TW202436339A - Use for treating cancer selected from non-small cell lung cancer or triple negative breast cancer - Google Patents

Abstract

This invention relates to the use of a bispecific antibody targeting programmed cell death protein 1 (PD-1) and lymphocyte activation gene-3 (LAG3) (PD1-LAG3) in the preparation of a medicament for treating a cancer selected from non-small cell lung cancer (NSCLC) and triple-negative breast cancer (TNBC) and its use in combination with (a) carboplatin and paclitaxel or (b) carboplatin and pemetrexed or (c) nab-paclitaxel.

Description

Use for treating cancer selected from non-small cell lung cancer or triple-negative breast cancer

The present invention relates to the use of a bispecific antibody (PD1-LAG3) targeting planned cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG3) in the preparation of a medicament for treating a cancer selected from non-small cell lung cancer (NSCLC) and triple-negative breast cancer. In particular, the present invention relates to methods and compositions for treating non-small cell lung cancer (NSCLC) in an individual by administering to the individual a bispecific antibody (PD-1-LAG3) targeting both programmed cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG3) and (a) carboplatin and paclitaxel or (b) carboplatin and pemetrexed, or the present invention relates to methods and compositions for treating triple-negative breast cancer (TNBC) in an individual by administering to the individual a bispecific antibody (PD1-LAG3) targeting both programmed cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG3) and nab-paclitaxel.

Lung cancer remains the leading cause of cancer death worldwide. NSCLC accounts for approximately 85% of all lung cancer cases. NSCLC can be divided into two subtypes: squamous (SQ) and non-squamous (NSQ). SQ histology accounts for approximately 25% of NSCLC. NSQ NSCLC includes several histologic subtypes, the most common of which is adenocarcinoma, which accounts for more than half of all NSCLC cases. The remainder of NSCLC cases are represented by other NSQ NSCLC histologies, including large cell carcinomas, neuroendocrine tumors, sarcomatoid carcinomas, and those with poorly differentiated histologies.

In the first-line setting, the standard of care for patients with advanced NSQ NSCLC is largely driven by the results of molecular profiling. For patients whose tumors harbor oncogenic driving mutations, the preferred first-line treatment utilizes approved targeted therapies, when available. For patients whose tumors lack targetable oncogenic aberrations, current standard of care in the first-line setting typically consists of immune checkpoint inhibitors (CPIs), including programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) blocking antibodies, with or without platinum doublet chemotherapy and bevacizumab. The current standard of care for newly diagnosed patients with advanced SQ NSCLC includes paclitaxel or gemcitabine in combination with a platinum agent.

Despite the remarkable progress in combination immunotherapy and the approval of immuno-CPI agents in combination with chemotherapy as first-line treatment for patients with advanced NSCLC, the majority of patients with advanced NSCLC progress during treatment and eventually experience disease progression and succumb to the disease. Therefore, there is still a large unmet medical need for patients with advanced NSCLC.

Breast cancer is the most commonly diagnosed cancer in women and the leading cause of cancer-related death in women worldwide. TNBC accounts for 12%-20% of newly diagnosed breast cancer cases. TNBC is immunohistologically characterized by the lack of expression of the hormones estrogen receptor (ER) and progesterone receptor (PgR), as well as the lack of overexpression and/or expansion of the human epidermal growth factor receptor 2 (HER2)/neurosaminidase ( NEU ) gene. Compared with other breast cancer subtypes, TNBC tumors are generally larger, more poorly differentiated, have more extensive lymph node involvement at diagnosis, and display an aggressive phenotype. Patients with TNBC have a higher risk of local and distant recurrence compared to patients with other breast cancers, and metastases are more likely to occur in internal organs and the brain rather than in bones. Patients with metastatic TNBC have a relatively poor prognosis (shorter progression-free survival (PFS) and overall survival (OS)) compared to patients with other breast cancer subtypes. In the setting of early-stage disease, this is reflected in a shorter time to recurrence and a shorter OS compared to patients with other breast cancers. Once breast cancer has metastasized, it is generally considered incurable.

In the first-line (1L) setting, treatment for this disease is chemotherapy, in combination with anti-planned death 1 (PD-1)/PD-L1 inhibitors (eg, atezolizumab, pembrolizumab) for patients with PD-L1-positive TNBC, where approved and readily available. In the 1L metastatic setting, OS remains modest at less than 3 years. Therefore, improvements in the combination of chemotherapy with PD-1/PD-L1-targeted agents remain urgently needed in 1L PD-L1-positive advanced TNBC. Combinations targeting novel immune checkpoints are attractive because they aim to exploit unique mechanisms to improve the success of immunotherapy in TNBC and have the potential to expand the proportion of patients whose tumors respond to immunotherapy.

In one aspect, the present disclosure provides a method for treating an individual with non-squamous non-small cell lung cancer (NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (b) pemetrexed; and (c) carboplatin.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the method comprises administering pemetrexed to the subject at a dose of 500 mg/m ² every three weeks.

In some aspects, the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and the method comprises administering to the individual the bispecific antibody, pemetrexed, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, the method comprises (a) administering to the individual a bispecific antibody first, administering pemetrexed second, and administering carboplatin third, or (b) administering to the individual a pemetrexed first, administering carboplatin second, and administering the bispecific antibody third.

In some aspects, the method comprises administering intravenously to the individual a bispecific antibody, pemetrexed, and carboplatin.

In some embodiments, (a) the bispecific antibody is administered over 60 (± 15) minutes in a first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) pemetrexed is administered over 10 minutes; and/or (d) carboplatin is administered over 30 to 60 minutes.

In some aspects, the dosing regimen comprises four dosing cycles.

In some aspects, the dosing regimen further comprises one or more additional dosing cycles of: (a) the bispecific antibody and (b) pemetrexed.

In some aspects, the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of: (a) the bispecific antibody and (b) pemetrexed.

In another aspect, the present disclosure provides a method for treating an individual having squamous NSCLC, the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (b) paclitaxel; and (c) carboplatin.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks.

In some aspects, the method comprises administering carboplatin to the subject at a target AUC of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and the method comprises administering to the individual the bispecific antibody, paclitaxel, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, the method comprises (a) administering to the individual the bispecific antibody first, administering paclitaxel second, and administering carboplatin third, or (b) administering to the individual paclitaxel first, administering carboplatin second, and administering the bispecific antibody third.

In some aspects, the method comprises administering intravenously to the individual a bispecific antibody, paclitaxel, and carboplatin.

In some aspects, (a) the bispecific antibody is administered over 60 (± 15) minutes in a first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) paclitaxel is administered over 3 hours; and/or (d) carboplatin is administered over 30 to 60 minutes.

In some aspects, the dosing regimen comprises four dosing cycles.

In some aspects, the dosing regimen further comprises one or more additional dosing cycles of the bispecific antibody.

In some aspects, the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody.

In some aspects, the subject is premedicated with oral or IV steroids and antihistamines prior to administration of paclitaxel.

In some aspects, the NSCLC is (a) stage IIIB/IIIC NSCLC; or (b) stage IV NSCLC.

In some embodiments, the individual has not received prior systemic therapy for metastatic NSCLC.

In some aspects, the individual has not been previously treated with immune checkpoint blockade therapy.

In some aspects, the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with an Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent.

In some embodiments, the individual has not been previously treated with a CD137 agonist.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or a mutation or anaplastic lymphoma kinase ( ALK ) fusion oncogene.

In some aspects, the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

In some aspects, a tumor sample of NSCLC from an individual has a PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TC) of <1%.

In some embodiments, the expression of PD-L1 on a PD-L1 TPS or TC in a tumor sample of NSCLC from an individual is between 1% and 49%.

In some aspects, the expression of PD-L1 on the PD-L1 TPS or TC in a tumor sample of NSCLC from the individual is ≥50%.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, (a) the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, pemetrexed, and carboplatin and does not comprise a bispecific antibody; or (b) the individual has squamous NSCLC and the control therapy comprises pembrolizumab, paclitaxel, and carboplatin and does not comprise a bispecific antibody.

In some embodiments, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising (i) a hypervariable region H1 (HVR-H1) sequence comprising an amino acid sequence of SEQ ID NO: 1, (ii) an HVR-H2 sequence comprising an amino acid sequence of GGR, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 3, (ii) an HVR-L2 sequence comprising an amino acid sequence of RSS, and (iii) an HVR-L3 sequence comprising SEQ ID NO: 4 The amino acid sequence.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. In some aspects, the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

In some aspects, the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. In some aspects, the Fc receptor is an Fcγ receptor.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7, (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12.

In some aspects, the first antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to the Kabat EU index number); and/or (b) an Fc domain comprising a modification that promotes the association of the first and second units of the Fc domain.

In some aspects, the first Fc domain unit comprises amino acid substitutions S354C and T366W (numbered according to Kabat EU index), and the second Fc domain unit comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to Kabat EU index).

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A, and P329G (according to Kabat EU index number).

In some aspects, in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other such that the VH domain is part of the light chain and the VL domain is part of the heavy chain.

In some aspects, in the first Fab fragment, the variable domains VL and VH are replaced with each other.

In some aspects, in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In another aspect, the present disclosure provides a method for treating an individual with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed, wherein the method comprises administering pemetrexed to the subject at a dose of 500 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL•min every three weeks.

In another aspect, the present disclosure provides a method for treating an individual with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising SEQ ID NO: and a second light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel, wherein the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL • min every three weeks.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with non-squamous non-small cell lung cancer (NSCLC), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; (b) pemetrexed; and (c) carboplatin.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the method comprises administering pemetrexed to the subject at a dose of 500 mg/m ² every three weeks.

In some aspects, the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and the method comprises administering to the individual the bispecific antibody, pemetrexed, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, the method comprises (a) administering to the individual a bispecific antibody first, administering pemetrexed second, and administering carboplatin third, or (b) administering to the individual a pemetrexed first, administering carboplatin second, and administering the bispecific antibody third.

In some aspects, the method comprises administering intravenously to the individual a bispecific antibody, pemetrexed, and carboplatin.

In some aspects, (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) pemetrexed is administered over 10 minutes; and/or (d) carboplatin is administered over 30 to 60 minutes.

In some aspects, the dosing regimen comprises four dosing cycles.

In some aspects, the dosing regimen further comprises one or more additional dosing cycles of: (a) the bispecific antibody and (b) pemetrexed.

In some aspects, the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of: (a) the bispecific antibody and (b) pemetrexed.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous NSCLC, wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; (b) paclitaxel; and (c) carboplatin.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks.

In some aspects, the method comprises administering carboplatin to the subject at a target AUC of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and the method comprises administering to the individual the bispecific antibody, paclitaxel, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, the method comprises (a) administering to the individual the bispecific antibody first, administering paclitaxel second, and administering carboplatin third, or (b) administering to the individual paclitaxel first, administering carboplatin second, and administering the bispecific antibody third.

In some aspects, the method comprises administering intravenously to the individual a bispecific antibody, paclitaxel, and carboplatin.

In some aspects, (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) paclitaxel is administered over 3 hours; and/or (d) carboplatin is administered over 30 to 60 minutes.

In some aspects, the dosing regimen comprises four dosing cycles.

In some aspects, the dosing regimen further comprises one or more additional dosing cycles of the bispecific antibody.

In some aspects, the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody.

In some aspects, the subject is premedicated with oral or IV steroids and antihistamines prior to administration of paclitaxel.

In some aspects, the NSCLC is (a) stage IIIB/IIIC NSCLC; or (b) stage IV NSCLC.

In some embodiments, the individual has not received prior systemic therapy for metastatic NSCLC.

In some aspects, the individual has not been previously treated with immune checkpoint blockade therapy.

In some embodiments, the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent.

In some embodiments, the individual has not been previously treated with a CD137 agonist.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or a mutation or anaplastic lymphoma kinase ( ALK ) fusion oncogene.

In some aspects, the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

In some aspects, a tumor sample of NSCLC from an individual has a PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TC) of <1%.

In some embodiments, the expression of PD-L1 on a PD-L1 TPS or TC in a tumor sample of NSCLC from an individual is between 1% and 49%.

In some aspects, the expression of PD-L1 on the PD-L1 TPS or TC in a tumor sample of NSCLC from the individual is ≥50%.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, (a) the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, pemetrexed, and carboplatin and does not comprise a bispecific antibody; or (b) the individual has squamous NSCLC and the control therapy comprises pembrolizumab, paclitaxel, and carboplatin and does not comprise a bispecific antibody.

In some embodiments, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising (i) a hypervariable region H1 (HVR-H1) sequence comprising an amino acid sequence of SEQ ID NO: 1, (ii) an HVR-H2 sequence comprising an amino acid sequence of GGR, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 3, (ii) an HVR-L2 sequence comprising an amino acid sequence of RSS, and (iii) an HVR-L3 sequence comprising SEQ ID NO: 4 The amino acid sequence.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. In some aspects, the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

In some aspects, the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. In some aspects, the Fc receptor is an Fcγ receptor.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7, (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12.

In some aspects, the first antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to the Kabat EU index number); and/or (b) an Fc domain comprising a modification that promotes the association of the first and second units of the Fc domain.

In some aspects, the first Fc domain unit comprises amino acid substitutions S354C and T366W (numbered according to Kabat EU index), and the second Fc domain unit comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to Kabat EU index).

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A, and P329G (according to Kabat EU index number).

In some aspects, in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. In some aspects, in the first Fab fragment, the variable domains VL and VH are replaced with each other.

In some aspects, in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (a) the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed, wherein the method comprises administering pemetrexed to the subject at a dose of 500 mg/ ^m2 every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL•min every three weeks.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (a) the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel, wherein the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL•min every three weeks.

In some aspects, the individual is a person.

In another aspect, the present disclosure provides a method for treating an individual with triple-negative breast cancer (TNBC), wherein the method comprises a dosing regimen comprising concurrently administering to the individual: (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antibody binding domain that specifically binds to LAG3; and (b) one or more dosing cycles of nab-paclitaxel.

In some embodiments, the individual has not previously received systemic anticancer therapy for locally advanced, unresectable, or metastatic TNBC.

In some aspects, the individual has not been previously treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the length of each of the one or more dosing cycles of the bispecific antibody is 21 days.

In some aspects, the method comprises administering a bispecific antibody to the individual on day 1 of each of the one or more dosing cycles.

In some embodiments, the method comprises administering the bispecific antibody intravenously to the subject. In some embodiments, (a) the bispecific antibody is administered over 60 (± 15) minutes in a first dosing cycle; and/or (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles.

In some aspects, the method comprises administering nab-paclitaxel to the individual at a dose of 100 mg/m ² once weekly for three weeks followed by one week off.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks; and administering nab-paclitaxel at a dose of 100 mg/m ² once weekly for three weeks, followed by one week off.

In some aspects, each of the one or more dosing cycles of nab-paclitaxel is 28 days in length.

In some aspects, the method comprises administering nab-paclitaxel to the subject on days 1, 8, and 15 of each of the one or more dosing cycles.

In some aspects, the method comprises administering nab-paclitaxel intravenously to the individual. In some aspects, the nab-paclitaxel is administered over 30 minutes.

In some aspects, on a day when the bispecific antibody and nab-paclitaxel are administered on the same day, the method comprises administering the bispecific antibody to the individual prior to nab-paclitaxel.

In some aspects, the TNBC is locally advanced, unresectable, or metastatic TNBC.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the TNBC is PD-L1 positive TNBC.

In some aspects, the PD-L1 positive TNBC has a PD-L1 combined positivity score (CPS) of ≥10 as measured using the pharmDx 22C3 IHC assay.

In some aspects, the PD-L1 positive TNBC has a PD-L1 tumor area positivity score (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay.

In some aspects, the PD-L1 positive TNBC has a PD-L1 combination immunoreductive (IC) ratio of ≥ 1% as measured using the Ventana SP142 IHC assay.

In some aspects, the dosing regimen comprises at least 5 or at least 10 dosing cycles of: (a) the bispecific antibody and (b) paclitaxel.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increased PFS rate at 12 months compared to a reference PFS rate at 12 months. In some aspects, the reference PFS rate is a PFS rate in a population of individuals who have received a control therapy.

In some aspects, the approach results in an increased OS rate at 12 months compared to a reference OS rate at 12 months. In some aspects, the reference OS rate is the OS rate in a population of individuals who have received a control therapy.

In some aspects, the control therapy comprises pembrolizumab and nab-paclitaxel and does not comprise a bispecific antibody.

In some aspects, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising (i) an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, (ii) an HVR-H2 sequence comprising the amino acid sequence GGR, and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, (ii) an HVR-L2 sequence comprising the amino acid sequence RSS, and (iii) an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. In some aspects, the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

In some aspects, the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. In some aspects, the Fc receptor is an Fcγ receptor.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7, (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12.

In some aspects, the first antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to the Kabat EU index number); and/or (b) an Fc domain comprising a modification that promotes the association of the first and second units of the Fc domain.

In some aspects, the first Fc domain unit comprises amino acid substitutions S354C and T366W (numbered according to Kabat EU index), and the second Fc domain unit comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to Kabat EU index).

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A, and P329G (according to Kabat EU index number).

In some aspects, in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. In some aspects, in the first Fab fragment, the variable domains VL and VH are replaced with each other.

In some aspects, in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In another aspect, the present disclosure provides a method for treating an individual with locally advanced, unresectable or metastatic TNBC, wherein the method comprises administering to the individual concurrently (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising SEQ ID NO: The method comprises administering nab-paclitaxel to the individual at a dose of 100 mg/m ² once a week for three weeks followed by one week of rest.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous triple-negative breast cancer (TNBC), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and nab-paclitaxel, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises concurrently administering to the individual (a) one or more dosing cycles of the bispecific antibody; and (b) one or more dosing cycles of nab-paclitaxel.

In some embodiments, the individual has not previously received systemic anticancer therapy for locally advanced, unresectable, or metastatic TNBC.

In some aspects, the individual has not been previously treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks.

In some aspects, the length of each of the one or more dosing cycles of the bispecific antibody is 21 days.

In some aspects, the method comprises administering a bispecific antibody to the individual on day 1 of each of the one or more dosing cycles.

In some embodiments, the method comprises administering the bispecific antibody intravenously to the subject. In some embodiments, (a) the bispecific antibody is administered over 60 (± 15) minutes in a first dosing cycle; and/or (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles.

In some aspects, the method comprises administering nab-paclitaxel to the individual at a dose of 100 mg/m ² once weekly for three weeks followed by one week off.

In some aspects, the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks; and administering nab-paclitaxel at a dose of 100 mg/m ² once weekly for three weeks, followed by one week off.

In some aspects, each of the one or more dosing cycles of nab-paclitaxel is 28 days in length.

In some aspects, the method comprises administering nab-paclitaxel to the subject on days 1, 8, and 15 of each of the one or more dosing cycles.

In some aspects, the method comprises administering nab-paclitaxel intravenously to the individual. In some aspects, the nab-paclitaxel is administered over 30 minutes.

In some aspects, on a day when the bispecific antibody and nab-paclitaxel are administered on the same day, the method comprises administering the bispecific antibody to the individual prior to nab-paclitaxel.

In some aspects, the TNBC is locally advanced, unresectable, or metastatic TNBC.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the TNBC is PD-L1 positive TNBC.

In some aspects, the PD-L1 positive TNBC has a PD-L1 combined positivity score (CPS) of ≥10 as measured using the pharmDx 22C3 IHC assay.

In some aspects, the PD-L1 positive TNBC has a PD-L1 tumor area positivity score (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay.

In some aspects, the PD-L1 positive TNBC has a PD-L1 combination immunoreductive (IC) ratio of ≥ 1% as measured using the Ventana SP142 IHC assay.

In some aspects, the dosing regimen comprises at least 5 or at least 10 dosing cycles of: (a) the bispecific antibody and (b) paclitaxel.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increased PFS rate at 12 months compared to a reference PFS rate at 12 months. In some aspects, the reference PFS rate is a PFS rate in a population of individuals who have received a control therapy.

In some aspects, the approach results in an increased OS rate at 12 months compared to a reference OS rate at 12 months. In some aspects, the reference OS rate is the OS rate in a population of individuals who have received a control therapy.

In some aspects, the control therapy comprises pembrolizumab and nab-paclitaxel and does not comprise a bispecific antibody.

In some aspects, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising (i) an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, (ii) an HVR-H2 sequence comprising the amino acid sequence GGR, and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, (ii) an HVR-L2 sequence comprising the amino acid sequence RSS, and (iii) an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. In some aspects, the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

In some aspects, the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. In some aspects, the Fc receptor is an Fcγ receptor.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7, (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12.

In some aspects, the first antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to the Kabat EU index number); and/or (b) an Fc domain comprising a modification that promotes the association of the first and second units of the Fc domain.

In some aspects, the first Fc domain unit comprises amino acid substitutions S354C and T366W (numbered according to Kabat EU index), and the second Fc domain unit comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to Kabat EU index).

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A, and P329G (according to Kabat EU index number).

In some aspects, in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. In some aspects, in the first Fab fragment, the variable domains VL and VH are replaced with each other.

In some aspects, in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering).

In some aspects, the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with locally advanced, unresectable or metastatic TNBC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and albumin-bound paclitaxel, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual (a) 600 mg every three weeks The method comprises administering to the subject one or more dosing cycles of a fixed dose of the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel, wherein the method comprises administering albumin-bound paclitaxel to the subject once a week at a dose of 100 mg/ ^m2 for three weeks, followed by one week of rest.

In some aspects, the individual is a person.

In another aspect, a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is provided for use in the manufacture of a medicament for treating an individual or a group of individuals suffering from non-squamous non-small cell lung cancer (NSCLC), wherein the bispecific antibody is formulated for simultaneous or separate administration in combination with (1) pemetrexed and (2) carboplatin, and wherein the treatment comprises administering to the individual or group of individuals a dosing regimen comprising one or more dosing cycles of the drug, pemetrexed, and carboplatin.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks.

In some aspects, pemetrexed is formulated for administration at a dose of 500 mg/m ² every three weeks. In some aspects, carboplatin is formulated for administration at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In some aspects, a use is provided, wherein the length of each of the one or more dosing cycles is 21 days, and wherein the treatment comprises administering a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, a use is provided wherein a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin are formulated for intravenous administration.

In some embodiments, a use is provided, wherein the treatment comprises: (a) administering a bispecific antibody targeting PD-1 and LAG3 over a period of 60 (± 15) minutes in a first dosing cycle and over a period of 30 (± 10) minutes in one or more additional dosing cycles; (b) administering pemetrexed over a period of 10 minutes; and/or (c) administering carboplatin over a period of 30 to 60 minutes.

In some embodiments, a use is provided, wherein the dosing regimen comprises four dosing cycles. In some embodiments, the dosing regimen further comprises one or more additional dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3 and (b) pemetrexed.

In some aspects, a use is provided, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of a bispecific antibody targeting PD-1 and LAG3 and pemetrexed.

In another aspect, a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is provided for use in the manufacture of a medicament for treating an individual or a group of individuals suffering from squamous NSCLC, wherein the bispecific antibody is formulated for simultaneous or separate administration in combination with (1) paclitaxel and (2) carboplatin, and wherein the treatment comprises administering to the individual or group of individuals a dosing regimen comprising one or more dosing cycles of the drug, paclitaxel, and carboplatin.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks.

In some aspects, paclitaxel is formulated for administration at a dose of 200 mg/m ² every three weeks. In some aspects, carboplatin is formulated for administration at a target AUC of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and wherein the treatment comprises administering a bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin on day 1 of each of the one or more dosing cycles.

In some aspects, a bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin are formulated for intravenous administration.

In some embodiments, a use is provided wherein the treatment comprises: (a) administering the bispecific antibody over a period of 60 (± 15) minutes in a first dosing cycle and over a period of 30 (± 10) minutes in one or more additional dosing cycles; (b) administering paclitaxel over a period of 3 hours; and/or (c) administering carboplatin over a period of 30 to 60 minutes.

In some embodiments, a use is provided wherein the dosing regimen comprises four dosing cycles. In some embodiments, the dosing regimen further comprises one or more additional dosing cycles of a bispecific antibody targeting PD-1 and LAG3. In some embodiments, the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of a bispecific antibody targeting PD-1 and LAG3.

In some aspects, a use is provided wherein the subject is premedicated with oral or IV steroids and an antihistamine prior to administration of paclitaxel.

In some aspects, a use is provided, wherein the NSCLC is: (a) Stage IIIB/IIIC NSCLC; or (b) Stage IV NSCLC.

In some aspects, uses are provided wherein the individual has not received prior systemic therapy for metastatic NSCLC.

In some aspects, uses are provided wherein the individual has not been previously treated with immune checkpoint blockade therapy.

In some embodiments, a use is provided wherein the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor having an Ig and a tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent. In some embodiments, the individual has not been previously treated with a CD137 agonist.

In some aspects, uses are provided wherein the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, uses are provided wherein the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or the anaplastic lymphoma kinase ( ALK ) fusion oncogene.

In some aspects, a use is provided wherein the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

In some aspects, a use is provided wherein a tumor sample of NSCLC from an individual has a PD-L1 tumor proportion score (TPS) or an expression level of PD-L1 on tumor cells (TC) of <1%.

In some aspects, a use is provided wherein the expression of PD-L1 on the PD-L1 TPS or TC of a tumor sample of NSCLC from an individual is between 1% and 49%.

In some aspects, a use is provided, wherein the expression of PD-L1 on PD-L1 TPS or TC in a tumor sample of NSCLC from an individual is ≥50%.

In some aspects, a use is provided wherein the treatment results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in an objective response rate (ORR) in a population of individuals treated according to the method compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, uses are provided wherein the treatment results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR of a population of individuals who have received a control therapy.

In some embodiments, a use is provided wherein: (a) the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, pemetrexed, and carboplatin and does not comprise a bispecific antibody; or (b) the individual has squamous NSCLC and the control therapy comprises pembrolizumab, paclitaxel, and carboplatin and does not comprise a bispecific antibody.

In another aspect, a use of a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 in the manufacture of a medicament for treating an individual or a population of individuals suffering from triple-negative breast cancer (TNBC), wherein the bispecific antibody is formulated for concurrent administration in combination with nab-paclitaxel, and wherein the treatment comprises administering to the individual or the population of individuals a dosing regimen comprising one or more dosing cycles of the drug and nab-paclitaxel.

In some aspects, the individual has not previously received systemic anticancer therapy for locally advanced, unresectable, or metastatic TNBC. In some aspects, the individual has not previously been treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks.

In some aspects, a use is provided, wherein the length of each of the one or more dosing cycles of the bispecific antibody is 21 days. In some aspects, the bispecific antibody targeting PD-1 and LAG3 is administered on day 1 of each of the one or more dosing cycles.

In some aspects, uses are provided wherein the treatment comprises intravenous administration of a bispecific antibody targeting PD-1 and LAG3.

In some embodiments, a use is provided, wherein the treatment comprises administering a bispecific antibody targeting PD-1 and LAG3 over 60 (± 15) minutes in a first dosing cycle; and/or administering a bispecific antibody targeting PD-1 and LAG3 over 30 (± 10) minutes in one or more additional dosing cycles. In some embodiments, the treatment comprises administering nab-paclitaxel at a dose of 100 mg/m ² once a week for three weeks, followed by 1 week off. In some embodiments, the treatment comprises administering a bispecific antibody targeting PD-1 and LAG3 at a fixed dose of 600 mg every three weeks; and administering nab-paclitaxel at a dose of 100 mg/m ² once a week for three weeks, followed by 1 week off.

In some aspects, a use is provided, wherein the length of each of the one or more dosing cycles of nab-paclitaxel is 28 days. In some aspects, the treatment comprises administering nab-paclitaxel on days 1, 8, and 15 of each of the one or more dosing cycles.

In some aspects, a use is provided, wherein the treatment comprises intravenously administering nab-paclitaxel. In some aspects, the nab-paclitaxel is administered over 30 minutes.

In some aspects, a use is provided wherein on a day when the bispecific antibody and nab-paclitaxel are administered on the same day, the treatment comprises administering the bispecific antibody to the individual prior to nab-paclitaxel.

In some aspects, a use is provided wherein the TNBC is locally advanced, unresectable or metastatic TNBC. In some aspects, the individual does not have any symptomatic, untreated or actively progressive central nervous system (CNS) metastases.

In some aspects, a use is provided, wherein the TNBC is PD-L1 positive TNBC. In some aspects, the PD-L1 positive TNBC has a PD-L1 combined positivity score (CPS) of ≥10 as measured using the pharmDx 22C3 IHC assay. In some aspects, the PD-L1 positive TNBC has a PD-L1 tumor area positivity score (TAP) of ≥5% as measured using the Ventana SP263 IHC assay. In some aspects, the PD-L1 positive TNBC has a PD-L1 combined immunoreductive (IC) ratio of ≥1% as measured using the Ventana SP142 IHC assay.

In some aspects, a use is provided, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3 and (b) nab-paclitaxel.

In some aspects, a use is provided wherein the treatment results in an increase in progression-free survival (PFS) compared to a reference PFS. In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in an objective response rate (ORR) in a population of individuals treated according to the method compared to a reference ORR. In some aspects, the reference ORR is the ORR in a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in duration of response (DOR) compared to a reference DOR. In some aspects, the reference DOR is the DOR of a population of individuals who have received a control therapy.

In some aspects, uses are provided, wherein the method results in an increase in OS compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in PFS rate at 12 months compared to a reference PFS rate at 12 months. In some aspects, the reference PFS rate is a PFS rate in a population of individuals who have received a control therapy.

In some aspects, a use is provided wherein the treatment results in an increase in OS rate at 12 months compared to a reference OS rate at 12 months. In some aspects, the reference OS rate is the OS rate in a population of individuals who have received a control therapy.

In some aspects, a use is provided, wherein the control therapy comprises pembrolizumab and nab-paclitaxel and does not comprise a bispecific antibody targeting PD-1 and LAG3.

In some embodiments, a method for manufacturing a medicament for treating non-squamous NSCLC, squamous NSCLC or TNBC is provided, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) a highly variable region H1 (HVR-H1) sequence comprising an amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising an amino acid sequence of GGR; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising SEQ ID NO: 3 ; (ii) HVR-L2 sequence, which comprises the amino acid sequence RSS; and (iii) HVR-L3 sequence, which comprises the amino acid sequence of SEQ ID NO: 4.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. In some aspects, the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain.

In some aspects, the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. In some aspects, the Fc receptor is an Fcγ receptor.

In some embodiments, a method for manufacturing a medicament for treating non-squamous NSCLC, squamous NSCLC or TNBC is provided, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10; (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11. NO: 11; and (iii) HVR-L3 sequence, which comprises the amino acid sequence of SEQ ID NO: 12.

In some aspects, the first antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the association of the first unit of the Fc domain with the second unit.

In some embodiments, the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A, and P329G (numbered according to Kabat EU index).

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In some aspects, the bispecific antibody targeting PD-1 and LAG3 is tobemstomig.

In some embodiments, a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is used in the manufacture of a medicament for treating non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC in an individual or a group of individuals, wherein the treatment comprises administering to the individual a dosing regimen comprising one or more of the following dosing cycles: (a) a bispecific antibody targeting PD-1 and LAG3, administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising SEQ ID NO: 16 an amino acid sequence of SEQ ID NO: 17; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed administered at a dose of 500 mg/m ² every three weeks; and (c) carboplatin administered at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In some embodiments, a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is used in the manufacture of a medicament for treating squamous stage IIIB/IIIC NSCLC or stage IV NSCLC in an individual or a group of individuals, wherein the treatment comprises administering to the individual a dosing regimen comprising one or more of the following dosing cycles: (a) a bispecific antibody targeting PD-1 and LAG3, administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising SEQ ID NO: 16 an amino acid sequence of SEQ ID NO: 17; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel, administered at a dose of 200 mg/m ² every three weeks; and (c) carboplatin, administered at a target AUC of 5 mg/mL • min every three weeks.

In some aspects, a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is used in the manufacture of a medicament for treating an individual or a group of individuals with locally advanced, unresectable or metastatic TNBC, wherein the treatment comprises administering to the individual the following dosing regimen in combination: (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3 administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising SEQ ID NO: 16 an amino acid sequence of SEQ ID NO: 17; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel administered once weekly at a dose of 100 mg/m ² for three weeks, followed by one week off.

In all these aspects, the individual is a person.

The present invention provides therapeutic methods and compositions for treating non-small cell lung cancer (NSCLC) or triple negative breast cancer (TNBC). Compositions, uses and kits involving such combinations and/or dosing regimens are also provided herein. I. Definitions

This article uses the following abbreviations: CDR Complementary decision area ORR Overall response rate/objective response rate CR Complete response OS Overall survival DNA DNA PD-1 Programmed Death 1 DCR Disease control rate PD-L1 Programmed death ligand 1 DOR Response duration PD-L2 Programmed death ligand 2 Fab Fragment antigen binding PFS Disease-free survival Fc Crystallizable fragment PR Partial response FFPE Formalin fixation and paraffin embedding Q2W Every 2 weeks FR frame Q3W Every 3 weeks HVR Highly variable area Q4W Every 4 weeks IHC Immunohistochemistry RNA RNA IV Intravenous SLD Sum of longest diameters NSCLC Non-small cell lung cancer TNBC Triple Negative Breast Cancer

As used herein, the term "about" refers to the usual error range of various values that are well known to those skilled in the art. In this article, reference to a value or parameter of "about" includes (and describes) aspects directed to the value or parameter itself. For example, a description of "about X" includes a description of "X".

The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners, thereby eliminating T cell dysfunction caused by signaling of the PD-1 signaling axis, resulting in restoration or enhancement of T cell function (e.g., proliferation, cytokine production and/or target cell killing). As used herein, PD-1 axis binding antagonists include PD-L1 binding antagonists, PD-1 binding antagonists, and PD-L2 binding antagonists. In some cases, a PD-1 axis binding antagonist includes a PD-L1 binding antagonist or a PD-1 binding antagonist. In a preferred aspect, the PD-1 axis binding antagonist is a PD-L1 binding antagonist.

The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with signal transduction caused by the interaction of PD-L1 with any one or more of its binding partners (such as PD-1 and/or B7-1). In some examples, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In a specific aspect, a PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and/or B7-1. In some examples, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or interfere with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners (e.g., PD-1 or B7-1). In one example, the PD-L1 binding antagonist reduces negative co-stimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes (through PD-L1-mediated signaling), thereby reducing the dysfunction of the dysfunctional T cells (e.g., enhancing effector responses to antigen recognition). In some cases, the PD-L1 binding antagonist binds to PD-L1. In some cases, the PD-L1 binding antagonist is an anti-PD-L1 antibody (e.g., an anti-PD-L1 antagonist antibody). Exemplary anti-PD-L1 antagonist antibodies include atezolizumab, MDX-1105, MEDI4736 (durvalumab), MSB0010718C (avelumab), SHR-1316, CS1001, envafolimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, cosibelimab, lodalimab (lodapolimab), FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, and HS-636. In some embodiments, the anti-PD-L1 antibody is atezolizumab, MDX-1105, MEDI4736 (durvalumab), or MSB0010718C (avelumab). In one embodiment, the PD-L1 binding antagonist is MDX-1105. In another embodiment, the PD-L1 binding antagonist is MEDI4736 (durvalumab). In another embodiment, the PD-L1 binding antagonist is MSB0010718C (avelumab). In other embodiments, the PD-L1 binding antagonist can be a small molecule, for example, GS-4224, INCB086550, MAX-10181, INCB090244, CA-170 or ABSK041, which can be administered orally in some examples. Other exemplary PD-L1 binding antagonists include AVA-004, MT-6035, VXM10, LYN192, GB7003 and JS-003. In a preferred embodiment, the PD-L1 binding antagonist is atezolizumab.

The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates or interferes with signal transduction caused by the interaction of PD-1 with one or more of its binding partners (such as PD-L1 and/or PD-L2). PD-1 (programmed death 1) is also known in the art as "programmed cell death 1", "PDCD1", "CD279" and "SLEB2". Exemplary human PD-1 is shown in UniProtKB/Swiss-Prot Accession No. Q15116. In some instances, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In a specific aspect, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and/or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, eliminate, or interfere with signal transduction caused by the interaction of PD-1 with PD-L1 and/or PD-L2. In one example, a PD-1 binding antagonist reduces negative co-stimulatory signals mediated by or expressed by cell surface proteins expressed on T lymphocytes (via PD-1 mediated signaling), thereby reducing dysfunction of dysfunctional T cells (e.g., enhancing effector responses to antigen recognition). In some instances, a PD-1 binding antagonist binds to PD-1. In some instances, a PD-1 binding antagonist is an anti-PD-1 antibody (e.g., an anti-PD-1 antagonist antibody). Exemplary anti-PD-1 antagonist antibodies include nivolumab, pembrolizumab, MEDI-0680, PDR001 (spartalizumab), REGN2810 (cemiplimab), BGB-108, prolgolimab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, retifanlimab, sasanlimab, penpulimab, CS1003, HLX10, SCT-I10A, zimberelimab, batilimab In one embodiment, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another embodiment, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In another embodiment, the PD-1 binding antagonist is a PD-L2 Fc fusion protein, e.g., AMP-224. In another embodiment, the PD-1 binding antagonist is MED1-0680. In another embodiment, the PD-1 binding antagonist is PDR001 (spartalizumab). In another embodiment, the PD-1 binding antagonist is REGN2810 (simiprilimab). In another embodiment, the PD-1 binding antagonist is BGB-108. In another embodiment, the PD-1 binding antagonist is proglitinomab. In another embodiment, the PD-1 binding antagonist is carrelizumab. In another embodiment, the PD-1 binding antagonist is sintilimab. In another embodiment, the PD-1 binding antagonist is tislelizumab. In another embodiment, the PD-1 binding antagonist is toripalimab. Other additional exemplary PD-1 binding antagonists include BION-004, CB201, AUNP-012, ADG104 and LBL-006. In a specific embodiment, the PD-1 binding antagonist antibody comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6.

The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates or interferes with signal transduction caused by the interaction of PD-L2 with any one or more of its binding partners, such as PD-1. PD-L2 (programmed death ligand 2) is also known in the art as "programmed cell death 1 ligand 2", "PDCD1LG2", "CD273", "B7-DC" and "PDL2". An exemplary human PD-L2 is shown in UniProtKB/Swiss-Prot Accession No. Q9BQ51. In some instances, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a specific aspect, a PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. Exemplary PD-L2 binding antagonists include anti-PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or interfere with signal transduction resulting from the interaction of PD-L2 with any one or more of its binding partners (such as PD-1). In one aspect, a PD-L2 binding antagonist reduces negative co-stimulatory signals mediated by or expressed by cell surface proteins expressed on T lymphocytes (via PD-L2-mediated signaling), thereby reducing dysfunction of dysfunctional T cells (e.g., enhancing effector responses to antigen recognition). In some aspects, a PD-L2 binding antagonist binds to PD-L2. In some aspects, a PD-L2 binding antagonist is an immunoadhesin. In other aspects, a PD-L2 binding antagonist is an anti-PD-L2 antagonist antibody.

The terms "programmed death ligand 1" and "PD-L1" herein refer to native sequence human PD-L1 polypeptides. Native sequence PD-L1 polypeptides are provided according to Uniprot Accession No. Q9NZQ7. For example, native sequence PD-L1 may have an amino acid sequence as set forth in Uniprot Accession No. Q9NZQ7-1 (isoform 1) (SEQ ID NO: 38). In another example, native sequence PD-L1 may have an amino acid sequence as set forth in Uniprot Accession No. Q9NZQ7-2 (isoform 2). In another example, native sequence PD-L1 may have an amino acid sequence as set forth in Uniprot Accession No. Q9NZQ7-3 (isoform 3). PD-L1 is also known in the art as “planned cell death 1 ligand 1,” “PDCD1LG1,” “CD274,” “B7-H,” and “PDL1.”

When referring to residues in the variable domains, the Kabat numbering system is generally used (approximately residues 1-107 for the light chain and residues 1-113 for the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest . 5th ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). When referring to residues in the constant region of the immunoglobulin heavy chain, the "EU numbering system" or "EU index" is generally used (e.g., the EU index reported by Kabat et al., supra). The "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody.

The term "cancer" refers to a disease caused by the uncontrolled division of abnormal cells in a part of the body. In one instance, the cancer is non-small cell lung cancer (NSCLC), e.g., squamous or non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous or non-squamous NSCLC. Cancer includes solid tumor cancers and non-solid tumor cancers and locally advanced or metastatic cancers (e.g., locally advanced or metastatic tumors). In one instance, the cancer is breast cancer, e.g., triple negative breast cancer (TNBC, e.g., locally advanced, unresectable or metastatic TNBC). More generally, examples of cancer include, but are not limited to, carcinomas, lymphomas, germ cell tumors, sarcomas, and leukemias or lymphoid malignancies. More specific examples of such cancers include, but are not limited to, urothelial carcinoma (UC), including locally advanced and metastatic UC (mUC), bladder cancer (e.g., muscle invasive bladder cancer (MIBC) and non-muscle invasive bladder cancer (NMIBC), e.g., BCG-refractory NMIBC), MIBC urothelial bladder cancer (UBC); kidney cancer or renal cancer (e.g., renal cell carcinoma (RCC)); urinary tract cancer; lung cancer, such as small cell lung cancer (SCLC), including extended stage SCLC (ES-SCLC), non-small cell lung cancer (NSCLC), including squamous NSCLC or non-squamous NSCLC, including locally advanced unresectable NSCLC (e.g., stage IIIB NSCLC), or recurrent or metastatic NSCLC (e.g., stage IV NSCLC), lung adenocarcinoma, or squamous cell carcinoma (e.g., epithelial squamous cell carcinoma (e.g., lung squamous cell carcinoma)); pancreatic cancer (e.g., pancreatic ductal adenocarcinoma (PDAC), e.g., metastatic PDAC); head and neck cancer (e.g., SCCHN, e.g., recurrent/metastatic PD-L1-positive SCCHN, and head and neck squamous cell carcinoma (HNSCC)); ovarian cancer (OC); esophageal cancer; peritoneal cancer; hepatocellular carcinoma; gastric cancer (GC) (e.g., gastroesophageal junction (GEJ) cancer) or gastric cancer, including gastrointestinal cancer and gastrointestinal stromal cancer; neuroglioblastoma; urinary tract cancer; liver cancer; breast cancer (e.g., HER2+ breast cancer and triple-negative breast cancer (TNBC (early TNBC) (eTNBC) that is estrogen receptor negative (ER-), progesterone receptor negative (PgR-), and HER2 negative (HER2-); prostate cancer, such as castration-resistant prostate cancer (CRPC); peritoneal cancer; hepatocellular carcinoma; gastric cancer or stomach cancer, including gastrointestinal cancer and gastrointestinal stromal cancer; pancreatic cancer (e.g., pancreatic ductal adenocarcinoma (PDAC)); neuroglioblastoma; cervical cancer (e.g., stage IVB, metastatic, recurrent, or persistent cervical cancer, such as metastatic and/or recurrent PD-L1-positive cervical cancer); ovarian cancer; liver cancer; colorectal cancer; rectal cancer; colorectal cancer (CRC; e.g., microsatellite stable (MSS) and microsatellite instability (MSI)-low CRC (MSI-low); endometrial or uterine cancer; salivary gland cancer; prostate cancer; vulvar cancer; thyroid cancer; liver cancer; anal cancer; penile cancer; melanoma, including superficial spreading melanoma, malignant lentigo melanoma, acral lenticular melanoma, and nodular melanoma; multiple myeloma and B-cell lymphoma (including low-grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate-grade/follicular NHL; intermediate-grade diffuse NHL; high-grade immunoblastic NHL; high-grade lymphoblastic NHL; high-grade small non-lytic cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); hairy cell leukemia; chronic myeloblastic leukemia (CML); post-transplant lymphoproliferative disorder (PTLD); and myelodysplastic syndrome (MDS), as well as abnormal vascular proliferation associated with nevus hamartomatosis, edema (such as that associated with brain tumors), Meigs syndrome, brain cancer, head and neck cancer, and related metastases.

In some instances, the cancer (e.g., NSCLC, e.g., squamous or non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous or non-squamous NSCLC) is a tumor having a tumor microenvironment comprising CD8+ T cells expressing LAG3.

In one instance, the cancer is breast cancer (e.g., triple-negative breast cancer (TNBC, e.g., locally advanced, unresectable or metastatic TNBC)).

In some cases, the cancer may be unresectable (e.g., unresectable locally advanced or metastatic cancer).

The term "tumor" refers to all proliferative cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive in this document.

As used herein, "tumor cell" refers to any tumor cell present in a tumor or a sample thereof. Tumor cells can be distinguished from other cells that may be present in a tumor sample, for example, stromal cells and tumor-infiltrating immune cells, using methods known in the art and/or described herein.

"Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Therefore, as a therapeutic concept, "tumor immunity" is "cured" when such evasion is weakened, and the tumor is recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

As used herein, "metastasis" refers to the spread of cancer from its original site to other sites in the body. Cancer cells can break away from the primary tumor, infiltrate the lymph and blood vessels, circulate in the blood, and grow (metastasize) to distant lesions in normal tissues elsewhere in the body. Metastasis can be local or distant. Metastasis is a secondary process that depends on tumor cells breaking away from the primary tumor, traveling through the bloodstream, and landing at a distant site. At the new site, these cells establish a blood supply and can grow to form a life-threatening mass. Both stimulatory and inhibitory molecular pathways within tumor cells modulate this behavior, and crosstalk between tumor cells and distant host cells is also important.

As used herein, "treatment" includes effective cancer treatment with an effective amount of a therapeutic agent (e.g., a bispecific antibody targeting PD-1 and LAG3) or a combination of therapeutic agents (e.g., a bispecific antibody targeting PD-1 and LAG3 and a chemotherapeutic agent, such as nab-paclitaxel). Treatment herein specifically includes adjuvant therapy, neoadjuvant therapy, non-metastatic cancer therapy (e.g., locally advanced cancer therapy), and metastatic cancer therapy. Treatment can be first-line therapy (e.g., the patient may not have been previously treated or has not received prior systemic therapy), or second-line or advanced therapy.

Herein, "effective amount" refers to an amount of a therapeutic agent (e.g., a bispecific antibody targeting PD-1 and LAG3) or a combination of therapeutic agents (e.g., a bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents, such as (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel) that achieves a therapeutic outcome. In some examples, an effective amount of a therapeutic agent or combination of therapeutic agents is an amount of the therapeutic agent or combination of therapeutic agents that achieves the following clinical endpoints: improved progression-free survival (PFS), improved PFS rate (e.g., improved PFS rate at 12 months), improved objective response rate (ORR), improved overall survival (OS), improved OS rate (e.g., improved OS rate at 12 months), improved pathological response rate (PRR), improved disease control rate (DCR), complete response (CR), pathological complete response (pCR), partial response (PR), improved survival (e.g., disease-free survival (DFS) and/or improved duration of response (DOR).

As used herein, "complete response" and "CR" refer to the disappearance of all target lesions.

As used herein, "partial response" or "PR" means a reduction of at least 30% in the sum of the longest diameters of target lesions (SLD) in the absence of a CR, using the baseline SLD before treatment as a reference.

As used herein, "disease progression" and "PD" refer to an increase of at least 20% in the sum of the SLD of target lesions relative to the sum of the smallest diameters at the previous time point (including baseline). The appearance of one or more new lesions may also be considered PD.

As used herein, "stable disease" or "SD" means neither shrinking to meet the requirements of PR nor increasing to meet the requirements of PD, with the minimum sum as reference.

As used herein, "Disease Control Rate" and "DCR" refer to the percentage of patients who have achieved CR, PR, and stable disease (SD). For example, DCR can be defined as the proportion of patients with SD ≥ 12 weeks or CR or PR, as determined by the investigator according to RECIST v1.1.

As used herein, "overall response rate," "objective response rate," and "ORR" may refer interchangeably to the sum of the CR rate and the PR rate. For example, an objective response may be defined as a CR or PR according to Response Evaluation Criteria in Entities (RECIST) v.1.1, as determined by investigator assessment and confirmed by repeat assessment ≥4 weeks after initial recording. In another example, ORR may be defined as the proportion of patients with two consecutive CRs or PRs ≥4 weeks apart as determined by the investigator according to RECIST v1.1.

As used herein, "progression-free survival" and "PFS" refer to the length of time that cancer does not progress during and after treatment. PFS can include the time a patient experiences a CR or PR and the time a patient has stable disease. For example, PFS can be defined as the time from the first study treatment to the first progression or death from any cause, whichever occurs first, as determined by the investigator according to RECIST v.1.1. In another example, PFS can be defined as the time from study entry to the first progression or death from any cause, whichever occurs first, as determined by the investigator according to RECIST v.1.1.

As used herein, "overall survival" and "OS" refer to the length of time a patient is alive from the date of diagnosis of a disease (e.g., cancer) or the date of initiation of treatment. For example, OS can be defined as the time from the first study treatment to death from any cause.

As used herein, the terms "duration of response" and "DOR" refer to the length of time from the documented tumor response to the date of disease progression or death from any cause, whichever occurs first. For example, DOR may be defined as the time from the first documented objective response to the first documented disease progression or death from any cause, whichever occurs first, as determined by the investigator according to RECIST v1.1.

As used herein, the term "chemotherapeutic agent" refers to a compound that can be used to treat cancer. In some aspects, the chemotherapeutic agent is pemetrexed, paclitaxel, or carboplatin. In some aspects, the chemotherapeutic agent is a combination of paclitaxel and carboplatin. In some aspects, the chemotherapeutic agent is a combination of pemetrexed and carboplatin. In some aspects, the chemotherapeutic agent is albumin-bound paclitaxel.

Examples of other chemotherapeutic agents include EGFR inhibitors (including small molecule inhibitors (e.g., erlotinib (TARCEVA®, Genentech/OSI Pharm.); PD 183805 (CI 1033, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-oxolinyl)propoxy]-6-quinazolinyl]-2-acrylamide dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-N-oxolinylpropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butyramide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolyl]-4-(dimethylamino)-2-butenylamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); and dual EGFR/HER2 tyrosine kinase inhibitors, such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6-[5-[[[2-methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine); tyrosine kinase inhibitors (e.g., EGFR inhibitors; small molecule HER2 tyrosine kinase inhibitors, such as TAK165 (Takeda); CP-724,714, oral selective inhibitors of the ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors, such as EKB-569 (available from Wyeth), which preferentially binds to EGFR but inhibits both cells that overexpress HER2 and cells that overexpress EGFR; PKI-166 (Novartis); pan-HER inhibitors, such as canertinib (CI-1033; Pharmacia); Raf-1 inhibitors, such as the antisense agent ISIS-5132 (ISIS Pharmaceuticals), which inhibits Raf-1 signaling; non-HER-targeted tyrosine kinase inhibitors, such as imatinib mesylate (GLEEVEC®, Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors, such as sunitinib (SUTENT®, Pfizer); VEGF receptor tyrosine kinase inhibitors, such as vatalanib (PTK787/ZK222584, Novartis/Schering AG); MAPK extracellular regulated kinase I inhibitor CI-1040 (Pharmacia); quinazolines, such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines, such as CGP 59326, CGP 60261, and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidines; curcumin (diferulylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrphostin containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to HER-encoding nucleic acids); quinoxalines (U.S. Patent No. 5,804,396); tyrphostin (U.S. Patent No. 5,804,396); ZD6474 (Astra Zeneca); PTK-787 (Novartis/Schering AG); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis/Lilly); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis/Schering AG); INC-1C11 (Imclone); and rapamycin (sirolimus, RAPAMUNE®); proteasome inhibitors, such as bortezomib (VELCADE®, Millennium Pharm.); disulfiram; epigallocatechin gallate; salinosporamide A; carfilzomib; 17-AAG (geldermycin geldanamycin); radicicol; lactate dehydrogenase A (LDH-A); fulvestrant (FASLODEX®, AstraZeneca); rituzol (FEMARA®, Novartis), finasunate (VATALANIB®, Novartis); oxaliplatin (ELOXATIN®, Sanofi); 5-FU (5-fluorouracil); folinic acid; lonafamib (SCH 66336); sorafenib (NEXAVAR®, Bayer Labs); AG1478, alkylating agents, such as thiotepa and CYTOXAN®; alkyl sulfonates, such as busulfan, improsulfan improsulfan and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa and uredopa; ethyleneimines and methylamelamines, including altretamine, triethylmelamine, triethylphosphatamide, triethylthiophosphatamide and trihydroxymethylmelamine; acetogenins (especially bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); bryostatin; callystatin; CC-1065 (including its adolesin derivative); adozelesin, carzelesin and bizelesin synthetic analogs); candidiasis (specifically candidiasis 1 and candidiasis 8); adrenocortical steroids (including prednisolone and prednisolone); cyproterone acetate; 5α-reductase (including finasteride and dutasteride); vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatin; aldesleukin, talc duocarmycin (including synthetic analogs, KW-2189 and CB1-TM1; eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards, such as chlorambucil, chlomaphazine, chlorophosphamide, estramustine, effulamide, methyldichloroethylamine, methyldichloroethylamine hydrochloride, myclobutanil, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, such as dichloroethylnitrosourea, chlorozotocin, fotemustine, lomustine, nimustine nimustine and ranimustine; antibiotics such as enediynes (e.g., calicheamicin, especially calicheamicin gamma 1 and calicheamicin omega 1); dynemicins, including dynemicin A; bisphosphonates such as clodronate; esperamicin; and neocarzinostatin chromophores and related chromoproteins (enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, cactinomycin, carabicin, carmosin caminomycin, carzinophilin, chromomycinis, dactinomycin, detorubicin, 6-diazo-5-oxo-L-norleucine, N-morpholino-doxorubicin, cyano-N-morpholino-doxorubicin, 2-N-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, miromycins such as miromycin C, mycophenolic acid, nogalamycin, oleamicin, pelocycin peplomycin, bofomycin, puromycin, quelamycin, rodorubicin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolism drugs, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-hydroxypurine, thiopurine thiamiprine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; adrenal anti-adrenal drugs, such as glutathione, mitotane, trilostane; folic acid supplements, such as folinic acid, leucovorin, folate, leucovorin, dapoxetine ... acid; acetylgluconolactone; aldehyde phosphamide glycoside; aminoacetyl propionic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; mushroom polysaccharide; lonidainine; maytansinoids, such as maytansine and ansamitocins; mitoguanidine mitoguazone; mitoxantrone; mopidamnol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazine; procarbazine; PSK® polysaccharide complex (JHS Natural Products); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; crescent toxins (especially T-2 toxin, verrucosin verracurin A, roridin A, and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; mechlorethamine phenylbutyric acid; GEMZAR® (gemcitabine); 6-thioguanine; hydroxypurine; methotrexate; etoposide (VP-16); ifosfamide; mitoxantrone; mitoxantrone novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; and pharmaceutically acceptable salts, acids, prodrugs and derivatives of any of the foregoing.

Chemotherapy agents also include (i) anti-hormonal agents that have a modulating or suppressive effect on the hormones of the tumor, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and FARESTON® (toremifine citrate); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazoles, aminoglutaramide, MEGASE® (megestrol acetate), AROMASIN® (exemestane; Pfizer), Formestanie, Fadrozole, RIVISOR® (vorozole), FEMARA® (ritozole; Novartis), and ARIMIDEX® (anastrozole; AstraZeneca); (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, triptorelin, medroxyprogesterone acetate, dienstilbestrol, premarin; (premarin), flumetrazolone, all trans-retinoic acid, fentanyl (fenretinide) and troxacitabine (1,3-dioxycytidine nucleoside analogs); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signaling pathways associated with abnormal cell proliferation, such as PKC-Alpha, Ralf and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (such as ANGIOZYME®) and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, such as ALLOVECTIN®, LEUVECTIN® and VAXID®; (ix) growth inhibitors, including vinca alkaloids (vincas) (e.g., vincristine and vinblastine), NAVELBINE® (vinorelbine), taxanes (e.g., paclitaxel, nab-paclitaxel and docetaxel), topoisomerase II inhibitors (e.g., adriamycin, epirubicin, daunorubicin, etoposide and bleomycin) and DNA alkylating agents (e.g., tamoxifen, prednisone, dacarbazine, methotrexate, cisplatin, methotrexate, 5-fluorouracil and ara-C); and (x) pharmaceutically acceptable salts, acids, prodrugs and derivatives of any of the foregoing.

As used herein, the term "cytotoxic agent" refers to any agent that is harmful to cells (e.g., causes cell death, inhibits proliferation, or otherwise impedes cell function). Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² , and radioisotopes of Lu); chemotherapeutic agents; enzymes and fragments thereof, such as nucleases; and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin, including fragments and/or variants thereof. Exemplary cytotoxic agents can be selected from anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, anti-metabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutics, pro-apoptotic agents, LDH-A inhibitors, fatty acid biosynthesis inhibitors, cell cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors and cancer metabolism inhibitors. In one example, the cytotoxic agent is a platinum-based chemotherapeutic agent (e.g., carboplatin or cisplatin). In one embodiment, the cytotoxic agent is an EGFR antagonist, for example, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (e.g., dexmedetomidine). In one embodiment, the cytotoxic agent is a RAF inhibitor, for example, a BRAF and/or CRAF inhibitor. In one embodiment, the RAF inhibitor is vemurafenib. In one embodiment, the cytotoxic agent is a PI3K inhibitor.

The term "patient" or "subject" refers to a human patient or subject. For example, the patient or subject can be an adult.

The term "antibody" herein specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (such as bispecific antibodies) and antibody fragments, as long as they exhibit the desired biological activity. In one example, the antibody is a full-length monoclonal antibody. In one case, the antibody is a bispecific antibody.

As used herein, the term IgG "isotype" or "subtype" refers to any subtype of immunoglobulins defined by the chemical and antigenic properties of its constant regions.

Antibodies (immunoglobulins) can be classified into different classes according to the amino acid sequences of their heavy chain constant domains. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of them can be further divided into subtypes (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, γ, ɛ, γ and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are known and generally described in, for example, Abbas et al . Cellular and Mol. Immunology , 4th edition (WB Saunders, Co., 2000). An antibody may be part of a larger fusion molecule formed by covalent or non-covalent association of the antibody with one or more other proteins or peptides.

The terms "full-length antibody", "intact antibody" and "whole antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, not as an antibody fragment as defined below. These terms refer to an antibody that includes an Fc region.

The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain that includes at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. In one aspect, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, antibodies produced by host cells may undergo post-translational cleavage of one or more, particularly one or two amino acids at the C-terminus of the heavy chain. Therefore, antibodies produced by host cells through the expression of a specific nucleic acid molecule encoding a full-length heavy chain may include a full-length heavy chain, or may include a cleavage variant of the full-length heavy chain. The last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447). Thus, the C-terminal lysine (Lys447) or C-terminal glycine (Gly446) and lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified, the amino acid sequence of the heavy chain including the Fc region is herein indicated to be free of the C-terminal lysine (Lys447). In one aspect, the heavy chain including the Fc region specified herein contained in the antibodies disclosed herein comprises an additional C-terminal glycine-lysine dipeptide (G446 and K447). In one aspect, the heavy chain including the Fc region specified herein contained in the antibodies disclosed herein comprises an additional C-terminal glycine residue (G446). In one aspect, the heavy chain comprising the Fc region specified herein contained in the antibodies disclosed herein comprises an additional C-terminal lysine residue (K447). In one embodiment, the Fc region comprises a single amino acid substitution N297A of the heavy chain. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or the constant region is according to the EU numbering system, also known as the EU index, as described in Kabat et al ., Sequences of Proteins of Immunological Interest , 5th edition Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

"Naked antibody" refers to an antibody that is not conjugated to a foreign moiety (e.g., a cytotoxic moiety) or a radiolabel. Naked antibodies can be present in pharmaceutical compositions.

The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies comprising the population are identical and/or bind to the same antigenic determinant, except for possible variant antibodies that contain naturally occurring mutations or that arise during the production of the monoclonal antibody preparation, such variants are generally present in small amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (antigenic determinants), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on the antigen. Therefore, the modifier "monoclonal" indicates that the characteristics of the antibody are obtained from a substantially homogeneous antibody population, and should not be interpreted as requiring the antibody to be produced by any particular method. For example, monoclonal antibodies intended for use according to the present invention can be produced by a variety of techniques, including but not limited to fusion tumor methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci.

As used herein, the term "hypervariable region" or "HVR" refers to the regions of the antibody variable domain whose sequences are highly variable and which determine the antigen binding specificity, such as the "complementary determining region" ("CDR").

Typically, antibodies include six CDRs: three in the VH (CDR-H1, CDR-H2, CDR-H3), and three in the VL (CDR-L1, CDR-L2, CDR-L3). As used herein, exemplary CDRs include: (a) highly variable loops present at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)); (b) CDRs present at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al. , Sequences of Proteins of Immunological Interest , 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and (c) antigen contacts are present at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)).

Unless otherwise indicated, CDRs are identified according to the method described by Kabat et al., supra. Those skilled in the art will appreciate that CDR names may also be identified according to the method described by Chothia, supra, McCallum, supra, or any other scientifically accepted nomenclature system.

"Framework" or "FR" refers to the variable domain residues outside the complementary determining regions (CDRs). The FR of a variable domain is usually composed of four FR domains: FR1, FR2, FR3, and FR4. Therefore, CDR and FR sequences usually appear in the following order in VH (or VL): FR1-CDR-H1(CDR-L1)-FR2-CDR-H2(CDR-L2)-FR3-CDR-H3(CDR-L3)-FR4.

The term "variable domain residue numbering as in Kabat" or "amino acid position numbering as in Kabat" and variants thereof refer to the numbering system used for the heavy chain variable domain or light chain variable domain in the Kabat et al. antibody compilation above . Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening or insertion of the FR or HVR of the variable domain. For example, the heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and an inserted residue after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbers of residues in a given antibody can be determined by aligning regions of sequence homology with a "standard" Kabat numbering sequence.

As used herein, the term "monospecific" antibody refers to an antibody having one or more binding sites, each binding site binding to the same epitope of the same antigen. As used herein, the term "bispecific" antibody means that the antibody is capable of specifically binding to at least two different antigens, such as two binding sites, each binding site is formed by a pair of antibody heavy chain variable domains (VH) and antibody light chain variable domains (VL), and the binding sites bind to different antigens or bind to different antigenic determinants on the same antigen. The bispecific antibody is in a 1+1 format. Other bispecific antibody formats are 2+1 format (comprising two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope) or 2+2 format (comprising two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). Typically, a bispecific antibody comprises two antigen binding sites, wherein each antigen binding site is specific for a different antigen. In one instance, a bispecific antibody is a bispecific antibody that comprises an Fc domain.

As used herein, in the context of an immunohistochemistry (IHC) assay (e.g., an IHC assay staining for PD-L1 using antibodies SP142, SP263, 22C3, or 28-8), the "proportion of PD-L1-positive tumor cells" is the percentage of live tumor cells that exhibit partial or complete membrane staining (excluding cytoplasmic staining) at any intensity relative to all live tumor cells present in the sample after staining the sample. Therefore, the proportion of PD-L1-positive tumor cells can be calculated using the PD-L1 IHC SP142 (Ventana) assay, for example, by the formula PD-L1-positive tumor cell proportion = (number of PD-L1-positive tumor cells)/(total number of PD-L1-positive and PD-L1-negative tumor cells), where PD-L1 cytoplasmic staining of tumor cells and all non-tumor cells (e.g., tumor-infiltrating immune cells, normal cells, necrotic cells, and debris) are excluded from evaluation and scoring. It should be understood that any given diagnostic PD-L1 antibody may correspond to a particular IHC assay protocol and/or scoring terminology that can be used to derive the proportion of PD-L1 positive tumor cells. For example, the proportion of PD-L1 positive tumor cells can be derived from tumor cell samples stained with SP263, 22C3, SP142, or 28-8 using the OPTIVIEW® assay performed on the Benchmark ULTRA, the EnVision Flex performed on the AutostainerLink 48, the OPTIVIEW® assay and expansion performed on the Benchmark ULTRA, or the EnVision Flex performed on the AutostainerLink 48, respectively.

As used herein, the "Ventana SP142 IHC assay" is performed according to the Ventana PD-L1 (SP142) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is incorporated herein by reference in its entirety.

As used herein, the "Ventana SP263 IHC assay" was performed according to the Ventana PD-L1 (SP263) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is incorporated herein by reference in its entirety.

As used herein, the "pharmDx 22C3 IHC assay" is performed according to the PD-L1 IHC 22C3 pharmDx package insert (Carpinteria, CA: Dako, Agilent Pathology Solutions), which is incorporated herein by reference in its entirety.

As used herein, the "pharmDx 28-8 IHC assay" is performed according to the PD-L1 IHC 28-8 pharmDx package insert (Carpinteria, CA: Dako, Agilent Pathology Solutions), which is incorporated herein by reference in its entirety.

The term "product leaflet" is used to refer to instructions customarily included in commercial packages of therapeutic products that contain information about the indications, usage, dosage, routes of administration, combination therapy, contraindications and/or warnings for the use of such therapeutic products.

As used herein, "in combination with" means administering a treatment modality in addition to another treatment modality, for example, a treatment regimen that includes administering a bispecific antibody targeting planned cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG3) and one or more chemotherapeutic agents (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel). Thus, "in combination with" means administering a treatment modality before, during, or after a patient is administered another treatment modality.

A drug that is administered "concurrently" with one or more other drugs is administered during a treatment cycle that occurs at the same time as the one or more other drugs, during the same treatment cycle, and/or on the same day of treatment, and, as appropriate, at the same time as the one or more other drugs. For example, for a cancer therapy given every 3 weeks, the concomitantly administered drugs may each be administered on Day 1 of the 3-week cycle. For a dosing regimen that includes dosing cycles of two or more drugs with different dosing frequencies (e.g., a dosing regimen that includes dosing cycles of: (a) a drug that is administered every three weeks and (b) a drug that is administered once weekly for three weeks followed by one week off), the dosing regimens are concurrent if the dosing regimens occur over the same time period (e.g., dosing cycles of two or more drugs with different dosing frequencies begin on the same day).

As used herein, the term "adverse event" or "AE" refers to any unfavorable and unexpected sign (including abnormal laboratory findings), symptom, or illness that is temporally associated with the use of a medical treatment or procedure and that may or may not be considered related to the medical treatment or procedure. Adverse events may be classified by a "grade" as defined by the National Cancer Institute Common Terminology Criteria for Adverse Events v4.0 or v5.0 (NIH CTCAE). In some aspects, the AE is a low-grade AE, e.g., a Grade 1 or 2 AE. Grade 1 includes AEs with no symptoms or mild symptoms. Grade 2 includes AEs that are moderate and limit age-appropriate instrumental activities of daily living (e.g., preparing meals, shopping for groceries or clothing) and AEs for which local or non-invasive intervention is indicated. In other instances, the AE is a high-grade AE, such as a Grade 3, 4, or 5 AE. In some instances, the AE is a Grade 3 or 4 AE. Grade 3 includes AEs that are severe or medically significant, but not immediately life-threatening, and indicate the need for hospitalization or prolonged hospitalization. Grade 4 includes AEs that have life-threatening consequences and indicate the need for urgent intervention. Grade 5 includes AEs that resulted in or were associated with death.

As used herein, the term "treatment-related AE" is an AE that is judged by the investigator to occur as a result of treatment, such as a bispecific antibody targeting PD-1 and LAG3 and/or one or more chemotherapeutic agents, such as (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel.

As used in this application, the term "valent" refers to the presence of a specified number of binding domains in an antigen binding molecule. Thus, the terms "bivalent", "tetravalent" and "hexavalent" refer to the presence of two binding domains, four binding domains and six binding domains, respectively, in an antigen binding molecule. A bispecific antibody according to the present invention is at least "bivalent" and may be "trivalent" or "multivalent" (e.g., "tetravalent" or "hexavalent"). In a particular aspect, an antibody of the present invention has two or more binding sites and is bispecific. That is, an antibody may be bispecific even in the presence of more than two binding sites (i.e., the antibody is trivalent or multivalent).

"Antibody fragment" refers to a molecule other than an intact antibody, which comprises a portion of an intact antibody that binds to an antigen bound by the intact antibody. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab') ₂ ; bifunctional antibodies, trifunctional antibodies, tetrafunctional antibodies, cross-Fab fragments, linear antibodies; single-chain antibody molecules (e.g., scFv); multispecific antibodies and single-domain antibodies formed from antibody fragments. For a review of certain antibody fragments, see Hudson et al., Nat Med 9, 129-134 (2003). For a general description of scFv fragments, see Plückthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994); see also WO 93/16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab')2 fragments that contain salvage receptor binding epitope residues and have increased in vivo half-life, see U.S. Patent No. 5,869,046. Bifunctional antibodies are antibody fragments having two antigen binding domains, which may be bivalent or bispecific, see, e.g., EP 404,097; WO 1993/01161; Hudson et al., Nat Med 9, 129-134 (2003); and Hollinger et al., Proc Natl Acad Sci USA 90, 6444-6448 (1993). Trifunctional antibodies and tetrafunctional antibodies are also described in Hudson et al., Nat Med 9, 129-134 (2003). Single domain antibodies are antibody fragments comprising all or part of the heavy chain variable domain of an antibody or all or part of the light chain variable domain of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516 B1). In addition, antibody fragments comprise single polypeptide chains having the characteristics of a VH domain (i.e., capable of assembling together with a VL domain into a functional antigen binding site) or a VL domain (i.e., capable of assembling together with a VH domain into a functional antigen binding site) and thereby providing the antigen binding properties of a full-length antibody. Antibody fragments can be produced by a variety of techniques, including but not limited to proteolytic digestion of intact antibodies as described herein and production in recombinant host cells (e.g., E. coli or bacteriophage).

Papain digestion of intact antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each of which contains a heavy chain variable domain and a light chain variable domain as well as a co-localized domain of the light chain and the first co-localized domain (CH1) of the heavy chain. Therefore, as used herein, the term "Fab fragment" refers to an antibody fragment comprising a light chain fragment, which comprises a VL domain and a co-localized domain (CL) of the light chain, and a VH domain and the first co-localized domain (CH1) of the heavy chain. Fab' fragments differ from Fab fragments in that a few residues are added to the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is a Fab' fragment in which the cysteine residue of the co-localized domain carries a free thiol group. Pepsin treatment produces a F(ab') ₂ fragment that has two antigen-binding sites (two Fab fragments) and a portion of the Fc region.

The term "cross-Fab fragment" or "xFab fragment" or "cross-Fab fragment" refers to a Fab fragment in which the variable regions or constant regions of the heavy and light chains are exchanged. Two different chain compositions of the cross-Fab molecule may exist and are included in the bispecific antibody of the present invention: on the one hand, the variable regions of the heavy and light chains of the Fab are exchanged, that is, the cross-Fab molecule comprises a peptide chain composed of the light chain variable region (VL) and the heavy chain constant region (CH1), and a peptide chain composed of the heavy chain variable region (VH) and the light chain constant region (CL). This interchanging Fab molecule is also called cross-Fab _(VLVH) . On the other hand, when the constant regions of the heavy chain and light chain of Fab are exchanged, the crossover Fab molecule contains a peptide chain composed of the heavy chain variable region (VH) and the light chain constant region (CL), and a peptide chain composed of the light chain variable region (VL) and the heavy chain constant region (CH1). This type of exchange Fab molecule is also called crossover Fab _(CLCH1) .

"Single-chain Fab fragment" or "scFab" is a polypeptide composed of an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL) and a linker, wherein the antibody domains and the linker have one of the following in the order from N-terminal to C-terminal direction: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CL-linker-VL-CH1 or d) VL-CH1-linker-VH-CL; and wherein the linker is a polypeptide of at least 30 amino acids, preferably between 32 and 50 amino acids. The single-chain Fab fragments are stabilized by the native disulfide bonds between the CL domain and the CH1 domain. In addition, these single-chain Fab molecules can be further stabilized by the insertion of cysteine residues (e.g., position 44 of the variable heavy chain and position 100 of the variable light chain according to Kabat numbering) to generate interchain disulfide bonds.

"Cross-linked Fab fragment" or "x-scFab" is a polypeptide composed of an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL) and a linker, wherein the antibody domains and the linker have one of the following in the order from N-terminal to C-terminal direction: a) VH-CL-linker-VL-CH1 and b) VL-CH1-linker-VH-CL; wherein VH and VL together form an antigen-binding domain that specifically binds to an antigen, and wherein the linker is a polypeptide of at least 30 amino acids. In addition, these x-scFab molecules can be further stabilized by the insertion of cysteine residues (e.g., position 44 in the variable heavy chain and position 100 in the variable light chain according to Kabat numbering) to generate interchain disulfide bonds.

"Single-chain variable fragment (scFv)" is a fusion protein of the variable regions of the heavy chain ( _VH ) and light chain ( _VL ) of an antibody, connected to a short linker peptide of ten to about 25 amino acids. The linker is usually rich in glycine for flexibility and serine or threonine for solubility, and can connect the N-terminus of _VH to the C-terminus of _VL , or vice versa . Despite the removal of the constant regions and the introduction of the linker, this protein retains the specificity of the original antibody. scFv antibodies are described, for example, in Houston, JS, Methods in Enzymol. 203 (1991) 46-96). Furthermore, antibody fragments comprise single polypeptides which have the characteristics of a VH domain (ie, capable of assembling together with a VL domain into a functional antigen binding site) or a VL domain (ie, capable of assembling together with a VH domain into a functional antigen binding site) and thereby provide the antigen binding properties of a full-length antibody.

Single domain antibodies are antibody fragments composed of a single monomeric variable antibody domain. The first single domain is derived from the variable domain of the antibody heavy chain from camel (nanobody or _VHH fragment). In addition, the term single domain antibody includes the autonomous human heavy chain variable domain (aVH) or _VNAR fragments derived from shark. Fibronectin is a scaffold that can be engineered to bind to antigens. Fibronectin consists of a backbone of the natural amino acid sequence of the 10th domain of the 15 repeat units of type III human fibronectin (FN3). Three loops at one end of the β-sandwich structure can be engineered to enable Adnectins to specifically recognize therapeutic targets of interest. For further details, see Protein Eng. Des. Sel. 18, 435- 444 (2005), US20080139791, WO2005056764 and US6818418B1. Peptide aptamers are combinatorial recognition molecules composed of a constant scaffold protein (usually thioredoxin (TrxA)) containing a constrained variable peptide loop inserted at the active site. For further details, see Expert Opin. Biol. Ther. 5, 783–797 (2005). Microbodies are derived from naturally occurring mini-proteins of 25-50 amino acids in length that contain 3-4 cysteine bridges, examples of mini-proteins include KalataBI and conotoxins and knottins. Mini-proteins have loops that can be engineered to include up to 25 amino acids without affecting the overall folding of the mini-protein. For further details on engineered knottin domains, see WO2008098796.

The terms “a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3”, “a bispecific antibody that specifically binds to PD-1 and LAG3”, “a bispecific antibody specific for PD-1 and LAG3” or “anti-PD-1/anti-LAG3 antibody” are used interchangeably herein and refer to a bispecific antibody that is capable of binding to PD-1 and LAG3 with sufficient affinity such that the antibody can be used as a diagnostic and/or therapeutic agent targeting PD-1 and LAG3. In some aspects, a bispecific antibody targeting PD-1 and LAG3 comprises (a) a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 1, (ii) an HVR-H2 sequence comprising an amino acid sequence of GGR, and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 3, (ii) an HVR-L2 sequence comprising an amino acid sequence of RSS, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 4; and (b) A second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising (i) an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, (ii) an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9; and a VL domain comprising (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12. In some aspects, the first antigen-binding domain comprises: a VH domain comprising an amino acid sequence of SEQ ID NO: 5, and a VL domain comprising an amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising an amino acid sequence of SEQ ID NO: 13, and a VL domain comprising an amino acid sequence of SEQ ID NO: 14. In some aspects, the bispecific antibody that specifically binds to PD-1 and LAG3 is RO7247669. The RO7247669 antibody was first described in WO 2018/185043, which is hereby incorporated by reference in its entirety.

RO7247669 (or tobemstomig) is a novel fragment crystallizable (Fc)-silenced IgG1-based bispecific antibody (BsAb) in a 1+1 format that incorporates monovalent binding to the checkpoint receptors, PD-1 and lymphocyte activation gene 3 (LAG3). The use of a native IgG-like monovalent heterodimeric IgG1 format enables the antibody to bind to both PD-1 and LAG3 simultaneously. RO7247669 BsAb is engineered to preferentially bind to T cells that co-express both PD-1 and LAG3, or to a lesser extent, to regulatory T cells that express only LAG3. Monovalent binding to LAG3 reduces antibody (Ab) internalization after binding to the T cell surface, and when simultaneously bound to PD-1 and LAG3, RO7247669 is retained longer on the T cell surface. The PGLALA mutation has been introduced into the IgG1-based Fc region of RO7247669 to avoid drug shaving and, therefore, tumor-associated macrophage resistance mechanisms that have been observed with IgG4-based antibodies such as KEYTRUDA® (pembrolizumab) and OPDIVO® (nivolumab) (Arlauckas et al., Sci Transl Med , 9:eaal3604, 2017; Shen et al., Eur J Pharm Sci , 157:105629, 2021).

The term "PD-1", also known as planned cell death protein 1, is a 288 amino acid type I membrane protein first described in 1992 (Ishida et al., EMBO J., 11 (1992), 3887–3895). PD-1 is a member of the expanded CD28/CTLA-4 family of T cell regulators and has two ligands, PD-L1 (B7-H1, CD274) and PD-L2 (B7-DC, CD273). The structure of the protein includes an extracellular IgV domain, followed by a transmembrane region and an intracellular tail. The intracellular tail contains two phosphorylation sites, located within an immunoreceptor tyrosine-based inhibitory motif and within an immunoreceptor tyrosine-based switch motif, suggesting that PD-1 negatively regulates TCR signaling. This is consistent with the binding of the SHP-1 and SHP-2 phosphatases to the PD-1 cytoplasmic tail upon ligand binding. Although PD-1 is not expressed on naive T cells, it is upregulated upon T cell receptor (TCR)-mediated activation and has been observed on both activated and exhausted T cells (Agata et al., Int. Immunology 8 (1996), 765-772). These exhausted T cells have a dysfunctional phenotype and are unable to mount an appropriate response. Although PD-1 has a relatively broad expression pattern, its most important role may be as a co-inhibitory receptor on T cells (Chinai et al., Trends in Pharmacological Sciences 36 (2015), 587-595). Therefore, current treatment approaches focus on blocking the interaction between PD-1 and its ligands to enhance T cell responses. The terms "planned death 1", "planned cell death 1", "protein PD-1", "PD-1", "PD1", "PDCD1", "hPD-1" and "hPD-1" are used interchangeably and include variants, isoforms, species homologs of human PD-1, and analogs that share at least one common antigenic determinant with PD-1. The amino acid sequence of human PD-1 is shown in UniProt (www.uniprot.org) accession number Q15116 (SEQ ID NO: 19).

Supportive clinical data in the field of cancer immunotherapy have shown that therapies focused on enhancing T cell responses against cancer can result in significant survival benefits for patients with advanced malignant tumors (Hodi et al., N Engl J Med, 363: 711-723, 2010; Kantoff et al., N Engl J Med, 363: 411-422, 2010; Chen et al., Clin Cancer Res, 18: 6580-6587, 2012).

The PD-1/PD-L1 pathway serves as an immune checkpoint that temporarily suppresses immune responses in states of chronic antigenic stimulation, such as chronic infection or cancer. PD-1 is an inhibitory receptor expressed on activated and exhausted T cells, including tumor-infiltrating CD8 ⁺ T cells that recognize mutated tumor antigens (neoantigens). Binding of PD-L1 to PD-1 inhibits T cell proliferation, activation, cytokine production, and cytolytic activity, resulting in a functionally inactivated and exhausted T cell state (Butte et al., Immunity, 27: 111-122, 2007; Yang et al., J. Immunol., 187: 1113-1119, 2011). Therapeutic targeting of the PD-1/PD-L1 pathway to enhance anti-tumor T cell responses has been clinically validated across multiple solid tumors when used as a single agent and in combination with chemotherapy and other targeted agents.

As used herein, unless otherwise indicated, the term "LAG3" or "Lag-3" or "lymphocyte activation gene-3" or "CD223" refers to any native LAG3 from any vertebrate source, including mammals, such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed LAG3 as well as any form of LAG3 obtained in cell processing. The term also encompasses natural LAG3 variants, such as splice variants or allelic variants. In a preferred embodiment, the term "LAG3" refers to human LAG3. The amino acid sequence of an exemplary processed (signal-free) LAG3 is shown in SEQ ID NO: 20. The amino acid sequence of an exemplary extracellular domain (ECD) LAG3 is shown in SEQ ID NO: 21.

LAG3 is an immune checkpoint protein involved in the regulation of anti-tumor immunity and chronic infection. LAG3 is expressed on a subset of activated T cells, B cells, natural killer cells, and tolerogenic plasmacytoid dendritic cells, and is constitutively expressed on regulatory T cells (Huard et al., Immunogenetics, 39: 213-217, 1994). Structurally similar to CD4, LAG3 is a member of the Ig superfamily and binds to the class II major histocompatibility complex (MHC-II). The interaction between LAG3 and MHC-II inhibits T cell proliferation, activation, cytolytic function and the production of pro-inflammatory cytokines (Goldberg and Drake, Curr Top Microbiol Immunol, 344: 269-278, 2011).

LAG3 has been reported to be expressed across multiple tumor types, including NSCLC, hepatocellular carcinoma, breast cancer, ovarian cancer, melanoma, renal cell carcinoma, and prostate cancer, and is associated with poor prognosis (Matsuzaki et al., Proc Natl Acad Sci USA, 107: 7875-7880, 2010; Baitsch et al., J Clin Invest, 121: 2350-2360, 2011; Thommen et al., Cancer Immunol Res, 3: 1344-1355, 2015; He et al., Cancer Sci, 107: 1193-1197, 2016; Norström et al., Oncotarget, 7: 23581-23593, 2016). Clinical evaluation of anti-LAG3 agents, given as single agents or in combination with other CPIs, is ongoing in multiple early studies in patients with advanced solid tumors (Long et al., Genes Cancer, 9: 176-189, 2018). Preliminary data suggest that anti-LAG3 therapy is well tolerated as a single agent and in combination with anti-PD-1 therapy, with a safety profile consistent with other CPI therapies (Ascierto et al., Ann Oncol, 28(Suppl 5): mdx440.011, 2017; Hong et al., J Clin Oncol, 36: 3012, 2018; Stratton et al., SITC Abstract P325, 2018). Therefore, therapeutic targeting of LAG3 represents an attractive strategy for treating patients with NSCLC or TNBC.

The terms "anti-LAG3 antibody" and "antibody that binds to LAG3" refer to an antibody that is capable of binding to LAG3 with sufficient affinity such that the antibody can be used as a diagnostic and/or therapeutic agent targeting LAG3. In one aspect, the anti-LAG3 antibody binds to unrelated non-LAG3 proteins to an extent that is less than about 10% of the binding of the antibody to LAG3 as measured by, for example, a radioimmunoassay (RIA). In certain embodiments, the antibody binds to LAG3 with a dissociation constant (K _D ) of ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM (e.g., 10 ^-8 M or lower, e.g., 10 ^-8 M to 10 ^-13 M, e.g., 10 ^-9 M to 10 ^-13 M). In certain aspects, the anti-LAG3 antibody binds to an epitope of LAG3 that is conserved in LAG3 across different species. In a preferred embodiment, "anti-LAG3 antibody", "antibody that specifically binds to human LAG3" and "antibody that binds to human LAG3" refer to antibodies that specifically bind to human LAG3 antigen or its extracellular domain (ECD) with a binding affinity of _KD value of 1.0 x ^10-8 mol/l or less, in one embodiment, the _KD value is 1.0 x ^10-9 mol/l or less, in one embodiment, the _KD value is 1.0 x ^10-9 mol/l to 1.0 x ^10-13 mol/l. In this context, binding affinity is determined by standard binding assays, such as surface plasmon resonance technology (BIAcore®, GE-Healthcare Uppsala, Sweden), for example using the LAG3 extracellular domain. The term "anti-LAG3 antibody" also includes bispecific antibodies that are able to bind to LAG3 and a second antigen.

The "knob-and-hole" technique is described in, for example, US 5,731,168; US 7,695,936; Ridgway et al., Prot Eng 9, 617-621 (1996); and Carter, J Immunol Meth 248, 7-15 (2001). Generally, the method involves introducing a protrusion ("knob") at the interface of a first polypeptide and a corresponding cavity ("hole") in the interface of a second polypeptide so that the protrusion can be placed in the cavity, thereby promoting heterodimer formation and hindering homodimer formation. The protrusion is constructed by replacing smaller amino acid side chains on the interface of the first polypeptide with larger side chains (e.g., tyrosine or tryptophan). Complementary cavities of the same or similar size as the protrusions are formed in the interface of the second polypeptide by replacing larger amino acid side chains with smaller amino acid side chains (e.g., alanine or threonine). The protrusions and cavities can be made by altering the nucleic acid encoding the polypeptide (e.g., by mutagenesis directed to specific sites or by peptide synthesis). In a specific embodiment, the knob modification comprises the amino acid substitution T366W in one of the two subunits of the Fc domain, and the hole modification comprises the amino acid substitutions T366S, L368A, and Y407V in the other of the two subunits of the Fc domain. In another specific embodiment, the subunit comprising the Fc domain with a knob modification further comprises the amino acid substitution S354C, and the subunit comprising the Fc domain with a hole modification further comprises the amino acid substitution Y349C. The introduction of these two cysteine residues allows the formation of a disulfide bond between the two subunits of the Fc region, thereby further stabilizing the dimer (Carter, J Immunol Methods 248, 7-15 (2001)).

The term "effector function" refers to those biological activities attributed to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen-presenting cells, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

"Activating Fc receptors" are Fc receptors that, upon binding to the Fc region of an antibody, initiate a signaling event that stimulates the receptor-bearing cell to perform effector functions. Activating Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcαRI (CD89). A specific activating Fc receptor is human FcγRIIIa (see UniProt accession number P08637, version 141).

The term "peptide linker" refers to a peptide comprising one or more amino acids, typically about 2 to 20 amino acids. Peptide linkers are known in the art or described herein. Suitable non-immunogenic linker peptides are, for example, ( _G4S ) _n , ( _SG4 ) _n or _G4 ( _SG4 ) _n peptide linkers, wherein "n" is typically a number between 1 and 10 (typically between 2 and 4, in particular 2), i.e. a peptide selected from the group consisting of GGGGS (SEQ ID NO: 22), GGGGSGGGGS (SEQ ID NO: 23), SGGGGSGGGG (SEQ ID NO: 24) and GGGGSGGGGSGGGG (SEQ ID NO: 25), but also including the sequences GSPGSSSSGS (SEQ ID NO: 26), (G4S) ₃ (SEQ ID NO: 27), (G4S) ₄ (SEQ ID NO: 28), GSGSGSGS (SEQ ID NO: 29), GSGSGNGS (SEQ ID NO: 30). (SEQ ID NO: 30), GGSGSGSG (SEQ ID NO: 31), GGSGSG (SEQ ID NO: 32), GGSG (SEQ ID NO: 33), GGSGNGSG (SEQ ID NO: 34), GGNGSGSG (SEQ ID NO: 35) and GGNGSG (SEQ ID NO: 36). Peptide linkers of particular interest are (G4S) (SEQ ID NO: 22), ( _G4S ) ₂ or GGGGSGGGGS (SEQ ID NO: 23), (G4S) ₃ (SEQ ID NO: 27) and ( _G4S ) ₄ (SEQ ID NO: 29), more specifically ( _G4S ) ₂ (SEQ ID NO: 23) or GGGGSGGGGS (SEQ ID NO: 23).

"Fused to" or "linked to" means that the components (eg, antigen binding domain or Fc domain) are linked directly or via one or more peptide linkers via a peptide bond. II. Therapeutic methods and compositions for non-small cell lung cancer A. Non-squamous NSCLC

In one aspect, the present disclosure provides a method for treating an individual with non-squamous non-small cell lung cancer (NSCLC) (e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (b) pemetrexed; and (c) carboplatin.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with NSCLC (e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; (b) pemetrexed; and (c) carboplatin.

Exemplary bispecific antibodies targeting PD-1 and LAG3 are provided in Section IV below. A specific example of a bispecific antibody targeting PD-1 and LAG3 is PD1-LAG3 as defined herein. In some aspects, the method comprises administering the bispecific antibody (e.g., a bispecific antibody targeting PD-1 and LAG3 as provided in Section IV) at a fixed dose of 600 mg every three weeks (Q3W). In some aspects, the length of each of the one or more dosing cycles of the bispecific antibody is 21 days, and the method comprises administering (e.g., intravenously) the bispecific antibody to the individual on day 1 of each of the one or more dosing cycles. For example, the bispecific antibody can be administered by intravenous infusion over about 60 minutes or over about 30 minutes. In some aspects, the bispecific antibody is administered by intravenous infusion over a longer duration in a first cycle and over a shorter duration in a second or further dosing cycle: for example, in some aspects, the bispecific antibody is administered over 60 (± 15) minutes (e.g., over about 60 minutes) in a first dosing cycle and over 30 (± 10) minutes (e.g., over about 30 minutes) in one or more additional dosing cycles.

In some aspects, the method comprises administering (eg, intravenously) pemetrexed to the subject at a dose of 500 mg/m ² every three weeks.

In some aspects, the method comprises administering (e.g., intravenously) calcein to a subject at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and the method comprises administering (e.g., intravenously) the bispecific antibody, pemetrexed, and carboplatin to the subject on day 1 of each of the one or more dosing cycles.

On days when the bispecific antibody, pemetrexed, and carboplatin are administered on the same day, the method may comprise (a) administering the bispecific antibody to the individual first, pemetrexed second, and carboplatin third, or (b) administering pemetrexed to the individual first, carboplatin second, and the bispecific antibody third. Alternatively, the method may comprise (c) administering to the individual the bispecific antibody first, carboplatin second, and pemetrexed third; (d) administering to the individual carboplatin first, pemetrexed second, and the bispecific antibody third; (e) administering to the individual carboplatin first, the bispecific antibody second, and pemetrexed third; or (f) administering to the individual pemetrexed first, the bispecific antibody second, and carboplatin third.

In some embodiments, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles of the bispecific antibody, pemetrexed, and carboplatin. In some embodiments, the dosing regimen comprises four dosing cycles of the bispecific antibody, pemetrexed, and carboplatin.

In some aspects, the dosing regimen comprises one or more dosing cycles of the bispecific antibody, pemetrexed, and carboplatin (e.g., 4 dosing cycles), and further comprises one or more additional dosing cycles of (a) the bispecific antibody and (b) pemetrexed (e.g., one or more additional dosing cycles comprising administration of the bispecific antibody and pemetrexed as described above but not comprising administration of carboplatin). For example, in some aspects, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20, or more than 20 additional dosing cycles of (a) the bispecific antibody and (b) pemetrexed (e.g., comprises at least 5 or at least 10 additional dosing cycles of (a) the bispecific antibody and (b) pemetrexed).

For example, in one aspect, the present disclosure provides a method for treating an individual with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed, wherein the method comprises administering pemetrexed to the subject at a dose of 500 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks.

In one aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (a) the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed, wherein the method comprises administering pemetrexed to the subject at a dose of 500 mg/ ^m2 every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL•min every three weeks.

In some aspects, the non-squamous NSCLC is locally advanced unresectable or metastatic non-squamous NSCLC, for example, (a) stage IIIB/IIIC non-squamous NSCLC; or (b) stage IV non-squamous NSCLC.

In some aspects, non-squamous NSCLC (e.g., locally advanced unresectable or metastatic non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC) is first line (1L), e.g., the individual has not previously received systemic anticancer therapy for locally advanced, unresectable or metastatic NSCLC. In some aspects, the individual has not received prior systemic therapy for metastatic NSCLC.

In some aspects, the individual has not been previously treated with immune checkpoint blockade therapy.

In some aspects, the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with an Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent.

In some embodiments, the individual has not been previously treated with a CD137 agonist.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or a mutation or anaplastic lymphoma kinase ( ALK ) fusion oncogene.

In some aspects, the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

In some aspects, the non-squamous NSCLC is PD-L1 positive NSCLC. Exemplary methods for assessing PD-L1 expression are provided, e.g., in Section III below. In some aspects, the PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TCs) of a tumor sample from an individual with NSCLC is <1%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is between 1% and 49%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is ≥50%. In other cases, non-squamous NSCLC is PD-L1 negative NSCLC.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%), e.g., an increase of In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in objective response rate (ORR) in a population of subjects treated according to the method compared to a reference ORR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of In some aspects, the reference ORR is the ORR of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of 1%-5%, 5%-10%, 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95%, 95%-100%) compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in DOR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%) compared to a reference duration of response (DOR), e.g., an increase of In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, the subject has non-squamous NSCLC and the control therapy comprises pembrolizumab, pemetrexed, and carboplatin and does not comprise a bispecific antibody. For example, in some aspects, the control therapy consists of pembrolizumab, pemetrexed, and carboplatin.

In some aspects, the subject is a human. B. Squamous NSCLC

In another aspect, the present disclosure provides a method for treating an individual with squamous non-small cell lung cancer (NSCLC) (e.g., stage IIIB/IIIC or stage IV squamous NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (b) paclitaxel; and (c) carboplatin.

In one aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous NSCLC (e.g., stage IIIB/IIIC or stage IV squamous NSCLC), wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; (b) paclitaxel; and (c) carboplatin.

Exemplary bispecific antibodies targeting PD-1 and LAG3 are provided in Section IV below. A specific example of a bispecific antibody targeting PD-1 and LAG3 is PD1-LAG3 as defined herein. In some aspects, the method comprises administering the bispecific antibody (e.g., a bispecific antibody targeting PD-1 and LAG3 as provided in Section IV) at a fixed dose of 600 mg every three weeks (Q3W). In some aspects, the length of each of the one or more dosing cycles of the bispecific antibody is 21 days, and the method comprises administering (e.g., intravenously) the bispecific antibody to the individual on day 1 of each of the one or more dosing cycles. For example, the bispecific antibody can be administered by intravenous infusion over about 60 minutes or over about 30 minutes. In some aspects, the bispecific antibody is administered by intravenous infusion over a longer duration in a first cycle and over a shorter duration in a second or further dosing cycle: for example, in some aspects, the bispecific antibody is administered over 60 (± 15) minutes (e.g., over about 60 minutes) in a first dosing cycle and over 30 (± 10) minutes (e.g., over about 30 minutes) in one or more additional dosing cycles.

In some aspects, the method comprises administering (e.g., intravenously) paclitaxel to the individual at a dose of 200 mg/m ² every three weeks. In some aspects, the individual is premedicated (e.g., premedicated with oral or IV steroids and antihistamines prior to administration of paclitaxel).

In some aspects, the method comprises administering (e.g., intravenously) calcein to a subject at a target AUC of 5 mg/mL • min every three weeks.

In some aspects, the length of each of the one or more dosing cycles is 21 days, and wherein the method comprises administering to the individual the bispecific antibody, paclitaxel, and carboplatin on day 1 of each of the one or more dosing cycles.

On the day when the bispecific antibody, paclitaxel, and carboplatin are administered on the same day, the method may comprise (a) administering the bispecific antibody to the individual first, administering paclitaxel second, and administering carboplatin third, or (b) administering paclitaxel to the individual first, administering carboplatin second, and administering the bispecific antibody third. Alternatively, the method may comprise (c) administering the bispecific antibody to the individual first, administering carboplatin second, and administering paclitaxel third; (d) administering carboplatin to the individual first, administering paclitaxel second, and administering the bispecific antibody third; (e) administering carboplatin to the individual first, administering the bispecific antibody second, and administering paclitaxel third; or (f) administering paclitaxel to the individual first, administering the bispecific antibody second, and administering carboplatin third.

In some embodiments, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles of the bispecific antibody, paclitaxel, and carboplatin. In some embodiments, the dosing regimen comprises four dosing cycles of the bispecific antibody, paclitaxel, and carboplatin.

In some aspects, the dosing regimen comprises one or more dosing cycles of the bispecific antibody, paclitaxel, and carboplatin (e.g., 4 dosing cycles), and further comprises the bispecific antibody and one or more additional dosing cycles (e.g., one or more additional dosing cycles comprising administration of the bispecific antibody as described above but not comprising administration of paclitaxel or carboplatin). For example, in some aspects, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more than 20 additional dosing cycles of the bispecific antibody (e.g., comprises (a) at least 5 or at least 10 additional dosing cycles of the bispecific antibody.

For example, in one aspect, the present disclosure provides a method for treating an individual with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel, wherein the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL • min every three weeks.

In one aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3, paclitaxel, and carboplatin, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (a) the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel, wherein the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) carboplatin, wherein the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL•min every three weeks.

In some aspects, the squamous NSCLC is locally advanced unresectable or metastatic squamous NSCLC, for example, (a) stage IIIB/IIIC squamous NSCLC; or (b) stage IV squamous NSCLC.

In some aspects, squamous NSCLC (e.g., locally advanced unresectable or metastatic squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous NSCLC) is first line (1L), e.g., the individual has not previously received systemic anticancer therapy for locally advanced, unresectable or metastatic NSCLC. In some aspects, the individual has not received prior systemic therapy for metastatic NSCLC.

In some aspects, the individual has not been previously treated with immune checkpoint blockade therapy.

In some aspects, the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with an Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent.

In some embodiments, the individual has not been previously treated with a CD137 agonist.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or a mutation or anaplastic lymphoma kinase ( ALK ) fusion oncogene.

In some aspects, the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1.

In some aspects, the squamous NSCLC is PD-L1 positive NSCLC. Exemplary methods for assessing PD-L1 expression are provided, e.g., in Section III below. In some aspects, the PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TCs) of a tumor sample from an individual with NSCLC is <1%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is between 1% and 49%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is ≥50%. Among other forms, squamous NSCLC is PD-L1 negative NSCLC.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%), e.g., an increase of In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in objective response rate (ORR) in a population of subjects treated according to the method compared to a reference ORR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of In some aspects, the reference ORR is the ORR of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of 1%-5%, 5%-10%, 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95%, 95%-100%) compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in DOR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%) compared to a reference duration of response (DOR), e.g., an increase of In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, the subject has squamous NSCLC and the control therapy comprises pembrolizumab, paclitaxel, and carboplatin and does not comprise a bispecific antibody. For example, in some aspects, the control therapy consists of pembrolizumab, paclitaxel, and carboplatin.

In some aspects, the bispecific antibody achieves at least 90% LAG3 receptor occupancy (RO) in a tumor, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% RO in a tumor.

In some aspects, the individual is a human. C. Triple Negative Breast Cancer

In one aspect, the present disclosure provides a method for treating an individual with triple-negative breast cancer (TNBC) (e.g., locally advanced, unresectable or metastatic TNBC), the method comprising a dosing regimen comprising concurrently administering to the individual one or more dosing cycles of: (a) a bispecific antibody targeting planned cell death protein 1 (PD-1) and lymphocyte activation gene 3 (LAG3), comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, and (b) nab-paclitaxel.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with TNBC (e.g., locally advanced, unresectable or metastatic TNBC), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and nab-paclitaxel, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises concurrently administering to the individual (a) one or more dosing cycles of the bispecific antibody; and (b) one or more dosing cycles of nab-paclitaxel.

Exemplary bispecific antibodies targeting PD-1 and LAG3 are provided in Section IV below. A specific example of a bispecific antibody targeting PD-1 and LAG3 is PD1-LAG3 as defined herein. In some aspects, the method comprises administering a bispecific antibody (e.g., a bispecific antibody targeting PD-1 and LAG3 as provided in Section IV) at a fixed dose of 600 mg every three weeks (Q3W). In some aspects, the length of each of the one or more dosing cycles of the bispecific antibody is 21 days. In some aspects, the method comprises administering (e.g., intravenously) a bispecific antibody to the individual on day 1 of each of the one or more dosing cycles. For example, the bispecific antibody can be administered by intravenous infusion over about 60 minutes or over about 30 minutes. In some aspects, the bispecific antibody is administered by intravenous infusion over a longer duration in a first cycle and over a shorter duration in a second or further dosing cycle: for example, in some aspects, the bispecific antibody is administered over 60 (± 15) minutes (e.g., over about 60 minutes) in a first dosing cycle and over 30 (± 10) minutes (e.g., over about 30 minutes) in one or more additional dosing cycles.

In some aspects, the method comprises administering nab-paclitaxel to the individual at a dose of 100 mg/m ² once a week for three weeks followed by one week off. In some aspects, the length of each of the one or more dosing cycles of nab-paclitaxel is 28 days. In some aspects, the method comprises administering (e.g., administering intravenously) nab-paclitaxel to the individual on days 1, 8, and 15 of each of the one or more dosing cycles. For example, in some aspects, nab-paclitaxel is administered by intravenous infusion over about 30 minutes (e.g., over 30 minutes).

On days when the bispecific antibody and nab-paclitaxel are administered on the same day (e.g., on the first day of the dosing regimen), the bispecific antibody can be administered before nab-paclitaxel. Alternatively, in some aspects, nab-paclitaxel is administered before the bispecific antibody.

In some aspects, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles of the bispecific antibody (e.g., at least 5 or at least 10 dosing cycles of the bispecific antibody). In some aspects, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more dosing cycles of nab-paclitaxel. In some aspects, the dosing regimen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 14, 15, 16, 17, 18, 19, 20 or more than 20 dosing cycles (e.g., at least 5 or at least 10 dosing cycles) of (a) the bispecific antibody and (b) nab-paclitaxel.

Accordingly, in some aspects, the method comprises administering to the individual a bispecific antibody at a fixed dose of 600 mg every three weeks; and administering nab-paclitaxel at a dose of 100 mg/m ² once a week for three weeks, followed by 1 week off.

In some aspects, TNBC (e.g., locally advanced, unresectable, or metastatic TNBC) is first line (1L), e.g., the individual has not previously received systemic anticancer therapy for locally advanced, unresectable, or metastatic TNBC.

In some aspects, the individual has not been previously treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody.

In some aspects, the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases.

In some aspects, the TNBC is PD-L1 positive TNBC. Exemplary methods for assessing the amount of PD-L1 expression are provided, e.g., in Section III below. In some aspects, the PD-L1 positive TNBC has a PD-L1 combined positive score (CPS) of ≥ 10 as measured using the pharmDx 22C3 IHC assay; a PD-L1 tumor area positive score (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay; and/or a PD-L1 positive immune cell (IC) ratio of ≥ 1% as measured using the Ventana SP142 IHC assay. In other aspects, the TNBC is PD-L1 negative TNBC.

In a specific aspect, the present disclosure provides a method for treating an individual with locally advanced, unresectable or metastatic TNBC, wherein the method comprises administering to the individual (a) a dosing regimen of one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual both of the specific antibodies at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising SEQ ID NO: 17 NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel, wherein the method comprises administering albumin-bound paclitaxel to the individual once a week at a dose of 100 mg/ ^m2 for three weeks, followed by 1 week of rest.

In another aspect, the present disclosure provides a bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with locally advanced, unresectable or metastatic TNBC, wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and albumin-bound paclitaxel, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual (a) 600 mg every three weeks The method comprises administering to the subject one or more dosing cycles of a fixed dose of the bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel, wherein the method comprises administering albumin-bound paclitaxel to the subject once a week at a dose of 100 mg/ ^m2 for three weeks, followed by one week of rest.

Recently, clinical proof of concept for dual inhibition of PD-1 and LAG3 was provided by the combination of relatlimab and nivolumab in patients with previously untreated metastatic or unresectable melanoma (Tawbi et al., N Engl J Med , 386(1): 24-34, 2022). By targeting both PD-1 and LAG-3 on dysfunctional tumor-specific T lymphocytes, bispecific antibodies targeting PD-1 and LAG3 aim to restore effective anti-tumor immune responses and provide even greater survival benefits to cancer patients compared to currently available checkpoint inhibitors. By preferentially targeting dysfunctional T cells co-expressing PD-1/LAG-3 and potentially reducing targeting of LAG-3-expressing Tregs in the tumor microenvironment, bispecific antibodies targeting PD-1 and LAG3 could avoid reinvigorating Treg-mediated immunosuppressive effects while restoring anti-tumor immune responses.

In some aspects, the method results in an increase in progression-free survival (PFS) compared to a reference PFS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%), e.g., an increase of In some aspects, the reference PFS is the PFS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in objective response rate (ORR) in a population of subjects treated according to the method compared to a reference ORR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of In some aspects, the reference ORR is the ORR of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in DOR (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%) compared to a reference duration of response (DOR), e.g., an increase of In some aspects, the reference DOR is the DOR for a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in OS (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of 1%-5%, 5%-10%, 10%-15%, 15%-20%, 20%-25%, 25%-30%, 30%-35%, 35%-40%, 40%-45%, 45%-50%, 50%-55%, 55%-60%, 60%-65%, 65%-70%, 70%-75%, 75%-80%, 80%-85%, 85%-90%, 90%-95%, 95%-100%) compared to a reference overall survival (OS). In some aspects, the reference OS is the OS of a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in PFS rate at 12 months (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of In some aspects, the reference PFS rate is the PFS rate for a population of individuals who have received a control therapy.

In some aspects, the method results in an increase in PFS rate at 12 months (e.g., an increase of at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or more than 100%, e.g., an increase of In some aspects, the reference PFS rate is the PFS rate for a population of individuals who have received a control therapy.

In some aspects, the control therapy comprises pembrolizumab and nab-paclitaxel and does not comprise a bispecific antibody. For example, in some aspects, the control therapy consists of pembrolizumab and nab-paclitaxel.

In some aspects, the bispecific antibody achieves at least 90% LAG3 receptor occupancy (RO) in a tumor, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% RO in a tumor.

In some aspects, the individual is a human. IV. Assessment of PD-L1 Expression

An individual treated according to any of the methods and compositions for use described herein may be assessed for PD-L1 expression. The methods and compositions for use may include determining the amount of PD-L1 expression in a biological sample (e.g., a tumor sample) obtained from an individual having cancer (e.g., NSCLC, e.g., squamous or non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous or non-squamous NSCLC, or TNBC, e.g., locally advanced, unresectable or metastatic TNBC). In other examples, the amount of PD-L1 expression in a biological sample (e.g., a tumor sample) obtained from an individual has been determined before starting treatment or after starting treatment. PD-L1 expression may be determined using any suitable method. For example, PD-L1 expression can be determined as described in U.S. Patent Publication Nos. US 20180030138 A1 and US 20180037655 A1. Any suitable tumor sample can be used, for example, a formalin-fixed and paraffin-embedded (FFPE) tumor sample, an archival tumor sample, a fresh tumor sample, or a frozen tumor sample.

In some aspects, the PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TC) of a cancer (e.g., NSCLC) or a sample therefrom is <1%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is between 1% and 49%. In some aspects, the PD-L1 TPS or PD-L1 expression on TCs of a tumor sample from an individual with NSCLC is ≥50%.

In some aspects, a cancer (e.g., TNBC) or a tumor sample therefrom is classified as PD-L1 positive if the cancer, or a tumor sample therefrom, has a PD-L1 combined positivity score (CPS) of ≥ 10 as measured using the pharmDx 22C3 IHC assay; a PD-L1 tumor area positivity score (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay; and/or a PD-L1 positive immune cell (IC) percentage of ≥ 1% as measured using the Ventana SP142 IHC assay. CPS is the number of PD-L1-staining cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells, multiplied by 100. IC is defined as the presence of discernible PD-L1 staining of any intensity in tumor-infiltrating immune cells covering the tumor area occupied by tumor cells, associated intratumoral stroma, and continuous peritumoral stroma. TAP is defined as the percentage of stained tumor and immune cells within the total tumor area.

For example, PD-L1 expression can be determined according to the percentage of tumor samples that are occupied by tumor-infiltrating immune cells that express a detectable amount of PD-L1 expression, as the percentage of tumor-infiltrating immune cells in a tumor sample that express a detectable amount of PD-L1 expression, and/or the percentage of tumor cells in a tumor sample that express a detectable amount of PD-L1 expression. It should be understood that in any of the foregoing examples, the percentage of a tumor sample occupied by tumor-infiltrating immune cells can be the percentage of tumor area covered by tumor-infiltrating immune cells in a tumor sample section obtained from an individual, for example, as assessed by IHC using an anti-PD-L1 antibody (e.g., SP142 antibody). Any suitable anti-PD-L1 antibody can be used, including, for example, SP142 (Ventana), SP263 (Ventana), 22C3 (Dako), 28-8 (Dako), E1L3N (Cell Signaling Technology), 4059 (ProSci, Inc.), h5H1 (Advanced Cell Diagnostics), and 9A11. In some examples, the anti-PD-L1 antibody is SP142. In other examples, the anti-PD-L1 antibody is SP263.

In some instances, a tumor sample obtained from an individual has detectable PD-L1 expression amount/level in less than 1% of tumor cells in the tumor sample, in 1% or more of tumor cells in the tumor sample, in 1% to less than 5% of tumor cells in the tumor sample, in 5% or more of tumor cells in the tumor sample, in 5% to less than 50% of tumor cells in the tumor sample, or in 50% to more of tumor cells in the tumor sample.

In some instances, a tumor sample obtained from an individual has detectable PD-L1 expression levels in tumor-infiltrating immune cells that comprise less than 1% of the tumor sample, more than 1% of the tumor sample, 1% to less than 5% of the tumor sample, more than 5% of the tumor sample, 5% to less than 10% of the tumor sample, or more than 10% of the tumor sample.

In some aspects, the cancer (e.g., NSCLC, e.g., squamous or non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous or non-squamous NSCLC, or TNBC, e.g., locally advanced, unresectable or metastatic TNBC) of an individual treated according to any of the methods provided herein has a PD-L1 positive tumor cell (TC) ratio or a tumor infiltrating immune cell (IC) ratio of <5%. In some aspects, esophageal cancer has a PD-L1 positive TC ratio of <1%. In other aspects, the cancer of an individual treated according to any of the methods provided herein has a PD-L1 positive TC ratio or IC ratio of ≥5%. In some aspects, PD-L1 is detected using a Ventana SP142 IHC assay, a Ventana SP263 IHC assay, a pharmDx 22C3 IHC assay, or a pharmDx 28-8 IHC assay.

In some examples, tumor samples can be scored for PD-L1 positivity in tumor-infiltrating immune cells and/or tumor cells according to the diagnostic assessment criteria shown in Table 1 and/or Table 2, respectively. Table 1. Tumor-Infiltrating Immune Cell (IC) IHC Diagnostic Criteria PD-L1 diagnostic assessment IC score Absence of any discernible PD-L1 staining or presence of any intensity of PD-L1 staining discernible in tumor-infiltrating immune cells covering <1% of the tumor area in tumor cells, associated intratumoral stroma, and contiguous peritumoral connective tissue proliferative stroma IC0 Presence of PD-L1 staining of any intensity discernible in tumor-infiltrating immune cells covering ≥1% to <5% of the tumor area in tumor cells, associated intratumoral stroma, and contiguous peritumoral connective tissue proliferative stroma IC1 Presence of PD-L1 staining of any intensity discernible in tumor-infiltrating immune cells covering ≥5% to <10% of the tumor area in tumor cells, associated intratumoral stroma, and contiguous peritumoral connective tissue proliferative stroma IC2 Presence of PD-L1 staining of any intensity discernible in tumor-infiltrating immune cells covering ≥10% of the tumor area in tumor cells, associated intratumoral stroma, and contiguous peritumoral connective tissue proliferative stroma IC3 Table 2. Tumor cell (TC) IHC diagnostic criteria PD-L1 diagnostic assessment TC score No discernible PD-L1 staining or discernible PD-L1 staining of any intensity in <1% of tumor cells TC0 PD-L1 staining of any intensity discernible in ≥1% to <5% of tumor cells TC1 PD-L1 staining of any intensity discernible in ≥5% to <50% of tumor cells TC2 PD-L1 staining of any intensity is discernible in ≥50% of tumor cells TC3 IV. Bispecific Antibodies Targeting PD-1 and LAG3 A. Exemplary Bispecific Antibodies Binding to PD -1 and LAG3

In one embodiment, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain comprising: (i) HVR-H1 comprising an amino acid sequence of GFSFSSY (SEQ ID NO: 1), (ii) HVR-H2 comprising an amino acid sequence of GGR, and (iii) HVR-H3 comprising an amino acid sequence of TGRVYFALD (SEQ ID NO: 2); and a VL domain comprising (i) HVR-L1 comprising an amino acid sequence of SESVDTSDNSF (SEQ ID NO: 3), (ii) HVR-L2 comprising an amino acid sequence of RSS, and (iii) HVR-L3, which comprises the amino acid sequence of NYDVPW (SEQ ID NO: 4).

In one aspect, the bispecific antibody comprises an Fc domain that is an IgG. In some aspects, the Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. In some aspects, the Fc domain has reduced or even eliminated effector function. In particular, the Fc domain may comprise one or more amino acid substitutions that reduce binding to an Fc receptor, in particular to an Fcγ receptor.

In another aspect, provided herein is a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the bispecific antibody comprises an Fc domain, the Fc domain is of IgG, specifically an IgG1 Fc domain or an IgG4 Fc domain, and wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor, specifically binding to an Fcγ receptor.

In another aspect, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain comprising: (i) HVR-H1 comprising an amino acid sequence of DYTMN (SEQ ID NO: 7), (ii) HVR-H2 comprising an amino acid sequence of VISWDGGGTYYTDSVKG (SEQ ID NO: 8), and (iii) HVR-H3 comprising an amino acid sequence of GLTDTTLYGSDY (SEQ ID NO: 9); and a VL domain comprising (i) HVR-L1 comprising an amino acid sequence of RASQSISSYLN (SEQ ID NO: 10), and (ii) HVR-L2, which comprises the amino acid sequence of AASTLQS (SEQ ID NO: 11), and (iii) HVR-L3, which comprises the amino acid sequence of QQTYSSPLT (SEQ ID NO: 12).

In one embodiment, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain comprising: (i) HVR-H1 comprising an amino acid sequence of GFSFSSY (SEQ ID NO: 1), (ii) HVR-H2 comprising an amino acid sequence of GGR, and (iii) HVR-H3 comprising an amino acid sequence of TGRVYFALD (SEQ ID NO: 2); and a VL domain comprising (i) HVR-L1 comprising an amino acid sequence of SESVDTSDNSF (SEQ ID NO: 3), (ii) HVR-L2 comprising an amino acid sequence of RSS; and The specifically binding second antigen-binding domain comprises: a VH domain comprising: (i) HVR-H1 comprising an amino acid sequence of DYTMN (SEQ ID NO: 7), (ii) HVR-H2 comprising an amino acid sequence of VISWDGGGTYYTDSVKG (SEQ ID NO: 8), and (iii) HVR-H3 comprising an amino acid sequence of GLTDTTLYGSDY (SEQ ID NO: 9); and a VL domain comprising (i) HVR-L1 comprising an amino acid sequence of RASQSISSYLN (SEQ ID NO: 10), (ii) HVR-L2 comprising an amino acid sequence of AASTLQS (SEQ ID NO: 11), and (iii) HVR-L3 comprising an amino acid sequence of QQTYSSPLT (SEQ ID NO: 12). In another aspect, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises a VH domain comprising an amino acid sequence of EVQLLESGGGLVQPGGSLRLSCAASGFSFSSYTMSWVRQAPGKGLEWVATISGGGRDIYYPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVLLTGRVYFALDSWGQGTLVTVSS (SEQ ID NO: 5); and a VL domain comprising The amino acid sequence of DIVMTQSPDSLAVSLGERATINCKASESVDTSDNSFIHWYQQKPGQSPKLLIYRSSTLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQNYDVPWTFGQGTKVEIK (SEQ ID NO: 6).

In another aspect, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the second antigen-binding domain that specifically binds to LAG3 comprises a VH domain comprising an amino acid sequence of EVQLLESGGGLVQPGGSLRL SCAASGFIFDDYTMNWVRQAPGKGLEWVAVISWDGGGTYYTDSVKGRFTISRDDFKNTLY LQMNSLRAEDTAVYYCAKGLTDTTLYGSDYWGQGTLVTVSS (SEQ ID NO: 13); and a VL domain comprising DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ The amino acid sequence of TYSSPLTFGGGTKVEIK (SEQ ID NO: 14).

In another aspect, the present invention provides a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain comprising the amino acid sequence of EVQLLESGGGLVQPGGSLRLSCAASGFSFSSYTMSWVRQAPGKGLEWVATISGGGRDIYYPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVLLTGRVYFALDSWGQGTLVTVSS (SEQ ID NO: 5); and a VL domain comprising DIVMTQSPDSLAVSLGERATINCKASESVDTSDNSFIHWYQQKPGQSPKLLIYRSSTLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQNYDVPWTFGQGTKVEIK (SEQ ID NO: 6); and the second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain comprising an amino acid sequence of EVQLLESGGGLVQPGGSLRLSCAASGFIFDDYTMNWVRQAPGKGLEWVAVISWDGGGTYYTDSVKGRFTISRDDFKNTLY LQMNSLRAEDTAVYYCAKGLTDTTLYGSDYWGQGTLVTVSS (SEQ ID NO: 13); and a VL domain comprising The amino acid sequence of DIQMTQSPSSSLSASVGDRVTITCRASQSISLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ TYSSPLTFGGGTKVEIK (SEQ ID NO: 14).

In one aspect, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain having at least 90% identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO:5; and a VL domain having at least 90% identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO:6. In one aspect, the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6.

In one aspect, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain having at least 90% identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO: 13; and a VL domain having at least 90% identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO: 14. In one aspect, the second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.

In one aspect, a bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain that has at least 90% identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO:5; and a VL domain that has at least 90% identity (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more than 99% identity) to the amino acid sequence of SEQ ID NO:6 and The second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain that is at least 90% identical (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical) to the amino acid sequence of SEQ ID NO: 13; and a VL domain that is at least 90% identical (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical) to the amino acid sequence of SEQ ID NO: 14.

In another aspect, a bispecific antibody is provided, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the first antigen-binding domain that specifically binds to PD-1 comprises: a VH domain comprising an amino acid sequence of SEQ ID NO: 5, and a VL domain comprising an amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain that specifically binds to LAG3 comprises: a VH domain comprising an amino acid sequence of SEQ ID NO: 13, and a VL domain comprising an amino acid sequence of SEQ ID NO: 14.

In a further aspect, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is a human antibody, a humanized antibody, or a chimeric antibody. Specifically, it is a humanized antibody or a chimeric antibody.

In one aspect, a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 is bivalent. This means that the bispecific antibody comprises one antigen-binding domain that specifically binds to PD-1 and one antigen-binding domain that specifically binds to LAG3 (1+1 format).

In one aspect, a bispecific antibody is provided, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the bispecific antibody comprises an Fc domain, a first Fab fragment comprising an antigen-binding domain that specifically binds to PD-1, and a second Fab fragment comprising an antigen-binding domain that specifically binds to LAG3. In a specific aspect, in one of the Fab fragments, the variable domains VL and VH are replaced with each other, such that the VH domain is part of the light chain and the VL domain is part of the heavy chain. In a specific aspect, in the first Fab fragment comprising an antigen-binding domain that specifically binds to PD-1, the variable domains VL and VH are replaced with each other.

In a specific aspect, a bispecific antibody is provided, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. For example, in one aspect, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18 (PD1-LAG3).

In another aspect, a bispecific antibody is provided, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the bispecific antibody comprises an Fc domain, a first Fab fragment comprising an antigen-binding domain that specifically binds to PD-1, and a second Fab fragment comprising an antigen-binding domain that specifically binds to LAG3, and the second Fab fragment is fused to the C-terminus of the Fc domain. Specifically, the Fab fragment comprising the antigen-binding domain that specifically binds to LAG3 is fused to the C-terminus of the Fc domain via its VH domain (trans 1+1 format).

In one aspect, the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 37; and a second light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 18. More specifically, the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 37; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18.

In a specific embodiment, the bispecific antibody targeting PD-1 and LAG3 is RO7247669 or tobemstomig. "Tobemstomig" is a bispecific antibody based on humanized IgG1, which comprises a monovalent binding to PD-1 and a monovalent binding to LAG3. The bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence of SEQ ID NO: 18. Tobemstomig is also described in WHO Drug Information (International Non-Proprietary Names of Drug Substances), Recommended INN: List 89, Volume 37, Issue 1, 2023 (page 203), see also Recommended INN: List 90, Volume 37, Issue 3, 2023, Corrigendum (page 907), and has the CAS registration number: 2648839-43-2. Fc domain modifications that reduce Fc receptor binding and / or effector function

In certain aspects, a bispecific antibody is provided, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the bispecific antibody comprises an Fc domain comprising one or more amino acid modifications that reduce binding to an Fc receptor, particularly an Fcγ receptor, and reduce or eliminate effector function.

In certain aspects, one or more amino acid modifications can be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant can comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g., substitution) at one or more amino acid positions.

The following sections describe preferred aspects of the bispecific antigen-binding molecules of the present invention, which comprise Fc domain modifications that reduce Fc receptor binding and/or effector function. In one aspect, the present invention relates to a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor, in particular to an Fcγ receptor. In particular, the Fc domain is of the human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index).

The Fc domain confers favorable pharmacokinetic properties to the bispecific antibodies of the present invention, including a long serum half-life and a favorable tissue-blood distribution ratio that facilitate good accumulation in target tissues. However, at the same time, it may cause the bispecific antibodies of the present invention to undesirably target cells expressing Fc receptors rather than preferred antigen-carrying cells. Accordingly, in specific embodiments, the Fc domain of the bispecific antibodies of the present invention exhibits reduced binding affinity to Fc receptors and/or reduced effector function compared to a native IgG Fc domain, specifically an IgG1 Fc domain or an IgG4 Fc domain. More specifically, the Fc domain is an IgG1 FC domain.

In one such aspect, the Fc domain (or the bispecific antigen-binding molecule of the invention comprising the Fc domain) exhibits less than 50%, preferably less than 20%, more preferably less than 10% and most preferably less than 5% of the binding affinity to the Fc receptor compared to a native IgG1 Fc domain (or a bispecific antigen-binding molecule of the invention comprising an IgG1 Fc domain), and/or exhibits less than 50%, preferably less than 20%, more preferably less than 10% and most preferably less than 5% of the effector function compared to a native IgG1 Fc domain (or a bispecific antigen-binding molecule of the invention comprising an IgG1 Fc domain). In one aspect, the Fc domain (or the bispecific antigen binding molecule of the invention comprising the Fc domain) does not substantially bind to an Fc receptor and/or induce effector function. In a specific aspect, the Fc receptor is an Fcγ receptor. In one aspect, the Fc receptor is a human Fc receptor. In one aspect, the Fc receptor is an activating Fc receptor. In a specific aspect, the Fc receptor is an activating human Fcγ receptor, more specifically human FcγRIIIa, FcγRI or FcγRIIa, most specifically human FcγRIIIa. In one aspect, the Fc receptor is an inhibitory Fc receptor. In a specific aspect, the Fc receptor is an inhibitory human Fcγ receptor, more specifically human FcγRIIB. In one aspect, the effector function is one or more of CDC, ADCC, ADCP, and cytokine secretion. In a specific aspect, the effector function is ADCC. In one aspect, the Fc domain exhibits substantially similar binding affinity to the neonatal Fc receptor (FcRn) compared to a native IgG1 Fc domain. Substantially similar binding to FcRn is achieved when the Fc domain (or a bispecific antigen-binding molecule of the invention comprising the Fc domain) exhibits greater than about 70%, particularly greater than about 80%, and more particularly greater than about 90% of the binding affinity of a native IgG1 Fc domain (or a bispecific antigen-binding molecule of the invention comprising an IgG1 Fc domain) to FcRn.

In certain aspects, the engineered Fc domain has reduced binding affinity to an Fc receptor and/or reduced effector function compared to a non-engineered Fc domain. In certain aspects, the Fc domain of the bispecific antigen-binding molecule of the present invention comprises one or more amino acid mutations that reduce the binding affinity of the Fc domain to an Fc receptor and/or effector function. Typically, the same one or more amino acid mutations are present in each of the two subunits of the Fc domain. In one aspect, the amino acid mutations reduce the binding affinity of the Fc domain to an Fc receptor. In another aspect, the amino acid mutations reduce the binding affinity of the Fc domain to an Fc receptor by at least 2-fold, at least 5-fold, or at least 10-fold. In one aspect, the bispecific antigen-binding molecules of the invention comprising an engineered Fc domain exhibit less than 20%, specifically less than 10%, more specifically less than 5% binding affinity to an Fc receptor compared to a bispecific antibody of the invention comprising a non-engineered Fc domain. In specific aspects, the Fc receptor is an Fcγ receptor. In other aspects, the Fc receptor is a human Fc receptor. In one aspect, the Fc receptor is an inhibitory Fc receptor. In specific aspects, the Fc receptor is an inhibitory human Fcγ receptor, more specifically human FcγRIIB. In some aspects, the Fc receptor is an activating Fc receptor. In a specific aspect, the Fc receptor is an activating human Fcγ receptor, more specifically human FcγRIIIa, FcγRI or FcγRIIa, most specifically human FcγRIIIa. Preferably, binding to each of these receptors is reduced. In some aspects, the binding affinity to complement components, specifically to C1q, is also reduced. In one aspect, the binding affinity to the neonatal Fc receptor (FcRn) is not reduced. Substantially similar binding to FcRn is achieved when the Fc domain (or the bispecific antigen-binding molecule of the invention comprising the Fc domain) exhibits greater than about 70% of the binding affinity of the non-engineered form of the Fc domain (or the bispecific antigen-binding molecule of the invention comprising the non-engineered form of the Fc domain) to FcRn, i.e., the binding affinity of the Fc domain to the receptor is maintained. The Fc domain or the bispecific antigen-binding molecule of the invention comprising the Fc domain may exhibit greater than about 80% and even greater than about 90% of this affinity. In certain embodiments, the Fc domain of the bispecific antigen-binding molecules of the invention is engineered to have reduced effector function compared to a non-engineered Fc domain. Reduced effector function may include, but is not limited to, one or more of the following: reduced complement-dependent cytotoxicity (CDC), reduced antibody-dependent cell-mediated cytotoxicity (ADCC), reduced antibody-dependent cellular phagocytosis (ADCP), reduced cytokine secretion, reduced immune complex-mediated antigen capture by antigen-presenting cells, reduced binding to NK cells, reduced binding to macrophages, reduced binding to monocytes, reduced binding to polymorphonuclear cells, reduced direct signaling that induces apoptosis, reduced dendritic cell maturation, or reduced T cell activation.

Antibodies with reduced effector function include those in which one or more of the Fc region residues 238, 265, 269, 270, 297, 327 and 329 are substituted (U.S. Patent No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" Fc mutant in which residues 265 and 297 are substituted with alanine (U.S. Patent No. 7,332,581). Certain antibody variants are described that have improved or reduced binding to FcRs. (e.g., U.S. Patent No. 6,737,056; WO 2004/056312; and Shields et al., J. Biol. Chem. 276 (2001) 6591-6604).

In one aspect of the invention, the Fc domain comprises amino acid substitutions at positions E233, L234, L235, N297, P331, and P329. In some aspects, the Fc domain comprises amino acid substitutions L234A and L235A ("LALA"). In one such embodiment, the Fc domain is an IgG1 Fc domain, particularly a human IgG1 Fc domain. In one aspect, the Fc domain comprises an amino acid substitution at position P329. In a more specific aspect, the amino acid substitution is P329A or P329G, particularly P329G. In one embodiment, the Fc domain comprises an amino acid substitution at position P329, and a further amino acid substitution selected from the group consisting of: E233P, L234A, L235A, L235E, N297A, N297D, or P331S. In a more specific embodiment, the Fc domain comprises amino acid mutations L234A, L235A, and P329G ("P329G LALA"). The "P329G LALA" combination of amino acid substitutions almost completely abolishes Fcγ receptor binding of human IgG1 Fc domains, as described in PCT Patent Application No. WO 2012/130831 A1. The document also describes methods for preparing such mutant Fc domains and methods for determining their properties (e.g. Fc receptor binding or effector function). Such antibodies are IgG1 with mutations L234A and L235A or with mutations L234A, L235A and P329G (EU index number according to Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition Public Health Service, National Institutes of Health, Bethesda, MD, 1991).

In one embodiment, the bispecific antibody of the invention comprises (all positions are according to the EU index numbering according to Kabat): (i) a homodimeric Fc region of human IgG1 subclass, optionally with the mutations P329G, L234A and L235A, or (ii) a homodimeric Fc region of human IgG4 subclass, optionally with the mutations P329G, S228P and L235E, or (iii) a homodimeric Fc region of human IgG1 subclass, optionally with the mutations P329G, L234A, L235A, I253A, H310A and H435A, or optionally with the mutations P329G, L234A, L235A, H310A, H433A and Y436A, or (iv) a heterodimeric Fc region wherein one Fc region polypeptide comprises the mutation T366W and the other Fc region polypeptide comprises the mutations T366S, L368A and Y407V, or wherein one Fc region polypeptide comprises the mutations T366W and Y349C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V and S354C, or wherein one Fc region polypeptide comprises the mutations T366W and S354C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V and Y349C, or (v) a heterodimeric Fc region of human IgG1 subclass The invention relates to an Fc region polypeptide comprising a polypeptide comprising the mutations P329G, L234A and L235A, and one Fc region polypeptide comprises the mutations T366W and the other Fc region polypeptide comprises the mutations T366S, L368A and Y407V, or one Fc region polypeptide comprises the mutations T366W and Y349C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V and S354C, or one Fc region polypeptide comprises the mutations T366W and S354C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V and Y349C.

In one aspect, the Fc domain is an IgG4 Fc domain. In a more specific aspect, the Fc domain is an IgG4 Fc domain comprising an amino acid substitution at position S228 (Kabat numbering), specifically an amino acid substitution S228P. In a more specific embodiment, the Fc domain is an IgG4 Fc domain comprising amino acid substitutions L235E and S228P and P329G. The amino acid substitutions reduce Fab group exchange of IgG4 antibodies in vivo (see Stubenrauch et al., Drug Metabolism and Disposition 38, 84-91 (2010)). Thus, in one aspect, a bispecific antibody is provided, comprising (all positions are numbered according to the EU index of Kabat): a heterodimeric Fc region of human IgG4 subclass, wherein both Fc region polypeptides comprise the mutations P329G, S228P and L235E, and one Fc region polypeptide comprises the mutations T366W and the other Fc region polypeptide comprises the mutations T366S, L368A and Y407V, or wherein one Fc region polypeptide comprises the mutations T366W and Y349C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V and S354C, or wherein one Fc region polypeptide comprises the mutations T366W and S354C and the other Fc region polypeptide comprises the mutations T366S, L368A, Y407V, and Y349C.

Antibodies with increased half-life and improved binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgG to the fetus (Guyer, R.L. et al., J. Immunol. 117 (1976) 587-593, and Kim, J.K. et al., J. Immunol. 24 (1994) 2429-2434), are described in US 2005/0014934. Those antibodies comprise an Fc region having one or more substitutions therein which improve binding of the Fc region to FcRn. Such Fc variants include those having substitutions at one or more of the Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424, or 434, e.g., substitution at Fc region residue 434 (U.S. Pat. No. 7,371,826). For other examples of Fc region variants, see also Duncan, A.R. and Winter, G., Nature 322 (1988) 738-740; US 5,648,260; US 5,624,821 and WO 94/29351.

Binding to Fc receptors can be readily determined by ELISA, or by surface plasmon resonance (SPR) using standard instrumentation such as a BIAcore instrument (GE Healthcare), and Fc receptors can be obtained, for example, by recombinant expression. Suitable such binding assays are disclosed herein. Alternatively, the binding affinity of an Fc domain or a cell-activating bispecific antigen binding molecule comprising an Fc domain to an Fc receptor can be assessed using a cell line known to express a specific Fc receptor, such as human NK cells expressing FcγIIIa receptors. The effector function of an Fc domain or a bispecific antibody of the invention comprising an Fc domain can be measured by methods known in the art. Assays suitable for measuring ADCC are described herein. Other examples of in vitro assays for assessing ADCC activity of molecules of interest are described in the following references: U.S. Patent No. 5,500,362; Hellstrom et al. Proc Natl Acad Sci USA 83, 7059-7063 (1986) and Hellstrom et al., Proc Natl Acad Sci USA 82, 1499-1502 (1985); U.S. Patent No. 5,821,337; Bruggemann et al., J Exp Med 166, 1351-1361 (1987). Alternatively, non-radioactive assays can be employed (see, e.g., ACTI™ Non-Radioactive Cytotoxicity Assay for Flow Cytometry (CellTechnology, Inc. Mountain View, CA); and CytoTox ^96® Non-Radioactive Cytotoxicity Assay (Promega, Madison, WI)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of a molecule of interest can be assessed in vivo in an animal model such as that disclosed by Clynes et al. in Proc Natl Acad Sci USA 95, 652-656 (1998).

The following section describes preferred aspects of the bispecific antibodies of the present invention, which comprise Fc domain modifications that reduce Fc receptor binding and/or effector function. In one aspect, the present invention relates to a bispecific comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, wherein the Fc domain comprises one or more amino acid substitutions that reduce the binding affinity of the antibody to an Fc receptor, particularly an Fcγ receptor. In another aspect, the present invention relates to a bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, wherein the Fc domain comprises one or more amino acid substitutions that reduce effector function. In a specific aspect, the Fc domain is of the human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index). Fc domain modifications that promote heterologous dimerization

The bispecific antigen-binding molecules of the present invention comprise different antigen-binding domains fused to one or the other of the two subunits of the Fc domain, so that the two subunits of the Fc domain can be contained in two different polypeptide chains. The recombinant co-expression of these polypeptides and the subsequent dimerization result in several possible combinations of the two polypeptides. In order to improve the yield and purity of the bispecific antibodies of the present invention in recombinant production, it would be advantageous to introduce modifications in the Fc domain of the bispecific antigen-binding molecules of the present invention that promote the binding of the desired polypeptides.

Thus, in a specific aspect, the present invention relates to a bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, wherein the Fc domain comprises a modification that promotes the first and second subunits of the Fc domain. The most extensive protein-protein interaction site between the two subunits of the human IgG Fc domain is in the CH3 domain of the Fc domain. Thus, in one aspect, the modification is performed in the CH3 domain of the Fc domain.

In a specific aspect, the modification is a so-called "knob-into-hole" modification, which comprises a "knob" modification in one of the two subunits of the Fc domain and a "hole" in the other of the two subunits of the Fc domain. Thus, the present invention relates to a bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding site that specifically binds to LAG3, wherein according to the knob-into-hole approach, the first subunit of the Fc domain comprises a knob and the second subunit of the Fc domain comprises a hole. In a specific aspect, the first subunit of the Fc domain comprises amino acid substitutions S354C and T366W (EU numbering), and the second subunit of the Fc domain comprises amino acid substitutions Y349C, T366S, and Y407V (according to Kabat EU index numbering).

The "knob-and-hole" technique is described in, for example, US 5,731,168; US 7,695,936; Ridgway et al., Prot Eng 9, 617-621 (1996); and Carter, J Immunol Meth 248, 7-15 (2001). Generally, the method involves introducing a protrusion ("knob") at the interface of a first polypeptide and a corresponding cavity ("hole") in the interface of a second polypeptide so that the protrusion can be placed in the cavity, thereby promoting heterodimer formation and hindering homodimer formation. The protrusion is constructed by replacing smaller amino acid side chains on the interface of the first polypeptide with larger side chains (e.g., tyrosine or tryptophan). By replacing larger amino acid side chains with smaller amino acid side chains (e.g., alanine or threonine), a complementary cavity of the same or similar size as the protrusion is formed in the interface of the second polypeptide.

Thus, in one aspect, in the CH3 domain of the first unit of the Fc domain of the bispecific antigen-binding molecule, an amino acid residue is substituted with an amino acid residue having a larger side chain volume, thereby generating a protrusion in the CH3 domain of the first unit, which can be positioned in the cavity in the CH3 domain of the second unit, and in the CH3 domain of the second unit of the Fc domain, an amino acid residue is substituted with an amino acid residue having a smaller side chain volume, thereby generating a cavity in the CH3 domain of the second unit, and the protrusion in the CH3 domain of the first unit can be positioned in the cavity. The protrusion and cavity can be prepared by altering the nucleic acid encoding the polypeptide (e.g., by mutation at a specific site or by peptide synthesis). In a specific embodiment, in the CH3 domain of the first unit of the Fc domain, the threonine residue at position 366 is replaced by a tryptophan residue (T366W), and in the CH3 domain of the second unit of the Fc domain, the tyrosine residue at position 407 is replaced by a valine residue (Y407V). In one embodiment, in addition in the second unit of the Fc domain, the threonine residue at position 366 is replaced by a serine residue (T366S), and the leucine residue at position 368 is replaced by an alanine residue (L368A).

In another embodiment, the serine residue at position 354 is replaced by a cysteine residue (S354C) in the first subunit of the Fc domain, and the tyrosine residue at position 349 is replaced by a cysteine residue (Y349C) in the second subunit of the Fc domain. The introduction of these two cysteine residues leads to the formation of a disulfide bridge between the two subunits of the Fc domain, further stabilizing the dimer (Carter (2001), J Immunol Methods 248, 7-15). In a specific aspect, the first subunit of the Fc domain comprises amino acid substitutions S354C and T366W (EU numbering), and the second subunit of the Fc domain comprises amino acid substitutions Y349C, T366S, and Y407V (according to Kabat EU index numbering).

However, alternatively or additionally, other knobs-in-hole techniques may be used as described in EP 1 870 459. In one embodiment, the multispecific antibody comprises the mutations R409D and K370E in the CH3 domain of the "knob chain" and the mutations D399K and E357K in the CH3 domain of the "hole chain" (numbering according to the Kabat EU index).

In one embodiment, the bispecific antibody comprises a T366W mutation in the CH3 domain of the "knob chain" and mutations T366S, L368A and Y407V in the CH3 domain of the "hollow chain", and additionally comprises mutations R409D and K370E in the CH3 domain of the "knob chain" and mutations D399K and E357K in the CH3 domain of the "hollow chain" (numbered according to the Kabat EU index).

In one embodiment, the bispecific antibody comprises mutations Y349C and T366W in one of the two CH3 domains and mutations S354C, T366S, L368A and Y407V in the other of the two CH3 domains; or the multispecific antibody comprises mutations Y349C and T366W in one of the two CH3 domains and mutations S354C, T366S, L368A and Y407V in the other of the two CH3 domains, and additionally comprises mutations R409D and K370E in the CH3 domain of the “knob chain” and mutations D399K and E357K in the CH3 domain of the “hole chain” (numbering according to the Kabat EU index).

In an alternative aspect, the modification that promotes the association of the first and second Fc domain subunits comprises a modification that mediates electrostatic switching, such as described in PCT Publication WO 2009/089004. Generally, this approach involves replacing one or more amino acid residues at the interface of two Fc domain subunits with charged amino acid residues, thereby making homodimer formation electrostatically unfavorable, but heterodimerization electrostatically favorable.

In addition to the "knobs-in-hole technique", other techniques for modifying the CH3 domain of the heavy chain of multispecific antibodies to enforce heterodimerization are known in the art. These techniques, in particular those described in WO 96/27011, WO 98/050431, EP 1870459, WO 2007/110205, WO 2007/147901, WO 2009/089004, WO 2010/129304, WO 2011/90754, WO 2011/143545, WO 2012/058768, WO 2013/157954 and WO 2013/096291 are contemplated herein as an alternative to the "knobs-in-hole technique" for use in combination with bispecific antibodies.

In one aspect, in a bispecific antibody, the method described in EP 1870459 is used to support heterodimerization of the first and second chains of the multispecific antibody. The method is based on the introduction of oppositely charged amino acids at specific amino acid positions at the CH3/CH3 domain interface between the two (i.e., the first and second chains).

Accordingly, in the aspect of the tertiary structure of the multispecific antibody, the CH3 domain of the first heavy chain and the CH3 domain of the second heavy chain form an interface between the CH3 domains of the corresponding antibodies, wherein the amino acid sequence of the CH3 domain of the first heavy chain and the amino acid sequence of the CH3 domain of the second heavy chain each comprise a set of amino acids within the interface within the tertiary structure of the antibody, wherein the set of amino acids within the interface within the CH3 domain of one heavy chain is substituted with positively charged amino acids, and the set of amino acids within the interface within the CH3 domain of the other heavy chain is substituted with negatively charged amino acids. Bispecific antibodies according to this aspect are referred to herein as "CH3(+/-)-engineered bispecific antibodies" (wherein the abbreviation "+/-" represents the oppositely charged amino acids introduced into the corresponding CH3 domains).

In one aspect, in the CH3(+/-) engineered bispecific antibody, the positively charged amino acid is selected from K, R and H, and the negatively charged amino acid is selected from E or D.

In one aspect, in the CH3(+/-) engineered bispecific antibody, the positively charged amino acid is selected from K and R, and the negatively charged amino acid is selected from E or D.

In one aspect, in the CH3(+/-) engineered bispecific antibody, the positively charged amino acid is K and the negatively charged amino acid is E.

In one embodiment, in a CH3(+/-) engineered bispecific antibody, amino acid R at position 409 in the CH3 domain of one heavy chain is substituted by D and amino acid K at position 409 is substituted by E, and amino acid D at position 399 in the CH3 domain of the other heavy chain is substituted by K and amino acid E at position 357 is substituted by K (according to Kabat EU index numbering).

In one aspect, the method described in WO 2013/157953 is used to support heterodimerization of the first and second heavy chains of the multispecific antibody. In one embodiment, the amino acid T at position 366 in the CH3 domain of one heavy chain is substituted with K, and the amino acid L at position 351 in the CH3 domain of the other heavy chain is substituted with D (numbered according to the Kabat EU index). In another embodiment, the amino acid T at position 366 in the CH3 domain of one heavy chain is substituted with K and the amino acid L at position 351 is substituted with K, and the amino acid L at position 351 in the CH3 domain of the other heavy chain is substituted with D (numbered according to the Kabat EU index).

In another aspect, the amino acid T at position 366 in the CH3 domain of one heavy chain is substituted by K and the amino acid L at position 351 is substituted by K, and the amino acid L at position 351 in the CH3 domain of the other heavy chain is substituted by D (according to the Kabat EU index numbering). In addition, the CH3 domain of the other heavy chain contains at least one of the following substitutions: the amino acid Y at position 349 is substituted by E, the amino acid Y at position 349 is substituted by D, and the amino acid L at position 368 is substituted by E (according to the Kabat EU index numbering). In one embodiment, the amino acid L at position 368 is substituted by E (according to the Kabat EU index numbering).

In one embodiment, the method described in WO 2012/058768 is used to support heterodimerization of the first and second chains of the multispecific antibody. In one embodiment, the amino acid L at position 351 in the CH3 domain of one chain is substituted by Y and the amino acid Y at position 407 is substituted by A, and the amino acid T at position 366 in the CH3 domain of the other chain is substituted by A and the amino acid K at position 409 is substituted by F (numbering according to the Kabat EU index). In another embodiment, in addition to the above substitutions, at least one of the amino acids at positions 411 (originally T), 399 (originally D), 400 (originally S), 405 (originally F), 390 (originally N), and 392 (originally K) in the CH3 domain of the other heavy chain is substituted (according to the Kabat EU index numbering). Preferred substitutions are: -     replacing the amino acid T at position 411 (numbered according to the Kabat EU index) with an amino acid selected from N, R, Q, K, D, E and W, -     replacing the amino acid D at position 399 (numbered according to the Kabat EU index) with an amino acid selected from R, W, Y and K, -     replacing the amino acid S at position 400 (numbered according to the Kabat EU index) with an amino acid selected from E, D, R and K, -     replacing the amino acid F at position 405 (numbered according to the Kabat EU index) with an amino acid selected from I, M, T, S, V and W; -     replacing the amino acid N at position 390 (numbered according to the Kabat EU index) with an amino acid selected from R, K and D; and -     replacing the amino acid S at position 400 (numbered according to the Kabat EU index) with an amino acid selected from The amino acids V, M, R, L, F and E are substituted for the amino acid K at position 392 (according to the Kabat EU index number).

In another aspect, the bispecific antibody is engineered according to WO 2012/058768, i.e., the amino acid L at position 351 in the CH3 domain of one heavy chain is substituted by Y and the amino acid Y at position 407 is substituted by V, and the amino acid T at position 366 in the CH3 domain of the other heavy chain is substituted by A and the amino acid K at position 409 is substituted by F (numbering according to Kabat EU index). In another embodiment of the multispecific antibody, amino acid Y at position 407 in the CH3 domain of one heavy chain is substituted by A, and amino acid T at position 366 in the CH3 domain of the other heavy chain is substituted by A and amino acid K at position 409 is substituted by F (numbering according to the Kabat EU index). In the last embodiment above, amino acid K at position 392 in the CH3 domain of the other heavy chain is substituted by E, amino acid T at position 411 is substituted by E, amino acid D at position 399 is substituted by R and amino acid S at position 400 is substituted by R (numbering according to the Kabat EU index).

In one embodiment, the method described in WO 2011/143545 is used to support heterodimerization of the first and second chains of the multispecific antibody. In one embodiment, amino acid modifications in the CH3 domains of the two chains are introduced at positions 368 and/or 409 (according to the Kabat EU index numbering).

In one aspect, the method described in WO 2011/090762 is used to support heterodimerization of the first and second chains of the bispecific antibody. WO 2011/090762 relates to amino acid modifications according to the "knobs-in-holes" (KiH) technique. In one embodiment, the amino acid T at position 366 in the CH3 domain of one heavy chain is substituted by W, and the amino acid Y at position 407 in the CH3 domain of the other heavy chain is substituted by A (numbering according to the Kabat EU index). In another embodiment, amino acid T at position 366 in the CH3 domain of one heavy chain is substituted with Y, and amino acid Y at position 407 in the CH3 domain of the other heavy chain is substituted with T (numbering according to the Kabat EU index).

In one aspect, the method described in WO 2009/089004 is used to support heterodimerization of the first and second chains of the bispecific antibody. In one embodiment, the amino acid K or N at position 392 in the CH3 domain of one heavy chain is substituted with a negatively charged amino acid (substituted with E or D in one embodiment, substituted with D in a preferred embodiment), and the amino acid D at position 399, the amino acid E or D at position 356, or the amino acid E at position 357 in the CH3 domain of the other heavy chain is substituted with a positively charged amino acid (substituted with K or R in one embodiment, substituted with K in a preferred embodiment; in a preferred embodiment, the amino acid at position 399 or 356 is substituted with K) (numbering according to the Kabat EU index). In another embodiment, in addition to the above substitutions, the amino acid K or R at position 409 in the CH3 domain of one heavy chain is substituted with a negatively charged amino acid (in one embodiment, substituted with E or D, in a preferred embodiment, substituted with D) (according to the Kabat EU index numbering). In yet another aspect, in addition to or in place of the above substitutions, the amino acid K at position 439 and/or the amino acid K at position 370 in the CH3 domain of one heavy chain are independently substituted with a negatively charged amino acid (in one embodiment, substituted with E or D, in a preferred embodiment, substituted with D) (according to the Kabat EU index numbering).

In one aspect, the method described in WO 2007/147901 is used to support heterodimerization of the first and second chains of the multispecific antibody. In one embodiment, the amino acid K at position 253 in the CH3 domain of one heavy chain is substituted by E, the amino acid D at position 282 is substituted by K and the amino acid K at position 322 is substituted by D, and the amino acid D at position 239 in the CH3 domain of the other heavy chain is substituted by K, the amino acid D at position 240 is substituted by K and the amino acid K at position 292 is substituted by D (numbering according to the Kabat EU index).

The C-terminus of the heavy chain of the bispecific antibody as reported herein can be a complete C-terminus ending with the amino acid residue PGK. The C-terminus of the heavy chain can be a shortened C-terminus, in which one or both C-terminal amino acid residues have been removed. In a preferred embodiment, the C-terminus of the heavy chain is a shortened C-terminus ending with PG. In one embodiment, the C-terminus of the heavy chain is a shortened C-terminus ending with P.

In one embodiment of all the embodiments reported herein, the bispecific antibody comprising a heavy chain comprising a C-terminal CH3 domain as specified herein comprises a C-terminal glycine-lysine dipeptide (G446 and K447, numbered according to the Kabat EU index). In one embodiment of all the embodiments reported herein, the bispecific antibody comprising a heavy chain comprising a C-terminal CH3 domain as specified herein comprises a C-terminal glycine residue (G446, numbered according to the Kabat EU index). i. Modifications in the Fab domain

In one embodiment, the present invention relates to a bispecific antibody comprising a first Fab fragment that specifically binds to PD-1 and a second Fab fragment that specifically binds to LAG3, wherein in one of the Fab fragments, the variable domains VH and VL or the constant domains CH1 and CL are exchanged. The bispecific antibody is prepared according to Crossmab technology.

Multispecific antibodies with domain swapping/interchanging in one binding pair (CrossMabVH-VL or CrossMabCH-CL) are described in detail in WO2009/080252, WO2009/080253 and Schaefer, W. et al., PNAS, 108 (2011) 11187-1191. They significantly reduce side products caused by mispairing of a light chain targeting a first antigen with an incorrect heavy chain targeting a second antigen (compared to approaches without such domain swapping).

In a specific embodiment, the present invention relates to a bispecific antibody comprising a first Fab fragment that specifically binds to PD-1 and a second Fab fragment that specifically binds to LAG3, wherein in one of the Fab fragments, the variable domains VL and VH are replaced with each other, such that the VH domain is part of the light chain and the VL domain is part of the heavy chain. In a specific embodiment, the bispecific antibody is a bispecific antibody in which, in the first Fab fragment comprising an antigen-binding domain that specifically binds to PD-1, the variable domains VL and VH are replaced with each other.

In another aspect, and to further improve correct pairing, a bispecific antibody comprising a first Fab fragment that specifically binds to PD-1 and a second Fab fragment that specifically binds to LAG3 may comprise different charged amino acid substitutions (so-called "charged residues"). These modifications are introduced into the cross-linked or non-cross-linked CH1 and CL domains. For example, such modifications are described in WO2015/150447, WO2016/020309 and PCT/EP2016/073408.

In a specific aspect, the present invention relates to a bispecific antibody comprising a first Fab fragment that specifically binds to PD-1 and a second Fab fragment that specifically binds to LAG3, wherein in one of the Fab fragments in the constant domain CL, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). In a specific aspect, the bispecific antibody is a bispecific antibody wherein, in the second Fab fragment comprising an antigen-binding domain that specifically binds to TIM3, the amino acid at position 124 in the constant domain CL is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index number), and the amino acids at positions 147 and 213 in the constant domain CH1 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index number).

In a specific aspect, the present invention relates to a bispecific antibody comprising a first Fab fragment that specifically binds to PD-1 and a second Fab fragment that specifically binds to LAG3, wherein in one of the CL domains, the amino acid at position 123 (EU numbering) has been replaced by arginine (R) and the amino acid at position 124 (EU numbering) has been replaced by lysine (K), and wherein in one of the CH1 domains, the amino acids at position 147 (EU numbering) and position 213 (EU numbering) have been substituted by glutamine (E). In a specific aspect, the bispecific antibody is a bispecific antibody in which, in the Fab fragment comprising the antigen-binding domain that specifically binds to LAG3, the amino acid at position 123 (EU numbering) has been replaced by arginine (R) and the amino acid at position 124 (EU numbering) has been replaced by lysine (K), and wherein in one of the CH1 domains, the amino acids at position 147 (EU numbering) and position 213 (EU numbering) have been substituted by glutamine (E).

In another embodiment, the bispecific antibody is a bivalent antibody comprising a) a first light chain and a first heavy chain of an antibody that specifically binds to a first antigen, and b) a second light chain and a second heavy chain of an antibody that specifically binds to a second antigen, wherein the variable domains VL and VH of the second light chain and the second heavy chain are replaced with each other.

The antibody under a) does not contain the modification reported under b), and the heavy chain and light chain under a) are separate chains.

In the antibody under b), the variable light chain domain VL within the light chain is replaced by the variable heavy chain domain VH of said antibody, and the variable heavy chain domain VH within the heavy chain is replaced by the variable light chain domain VL of said antibody.

In one embodiment, (i) the amino acid at position 124 (according to Kabat numbering) in the constant domain CL of the first light chain under a) is substituted with a positively charged amino acid, and wherein the amino acid at position 147 or the amino acid at position 213 (according to Kabat EU index numbering) in the constant domain CH1 of the first heavy chain under a) is substituted with a negatively charged amino acid, or (ii) the amino acid at position 124 (according to Kabat numbering) in the constant domain CL of the second light chain under b) is substituted with a positively charged amino acid, and wherein the amino acid at position 147 or the amino acid at position 213 (according to Kabat EU index numbering) in the constant domain CH1 of the second heavy chain under b) is substituted with a positively charged amino acid. The amino acid being treated is replaced by a negatively charged amino acid.

In another embodiment, (i) the amino acid at position 124 in the homeostatic domain CL of the first light chain under a) is independently substituted with lysine (K), arginine (R) or histidine (H) (according to Kabat numbering) (in a preferred embodiment, it is independently substituted with lysine (K) or arginine (R)), and wherein the amino acid at position 147 or the amino acid at position 213 in the homeostatic domain CH1 of the first heavy chain under a) is independently substituted with glutamine (E) or aspartic acid (D) (according to Kabat EU index numbering), or (ii) the amino acid at position 124 in the homeostatic domain CL of the second light chain under b) is independently substituted with lysine (K), arginine (R) or histidine (H) (according to Kabat EU index numbering). or histidine (H) (according to Kabat numbering) (in a preferred embodiment independently substituted by lysine (K) or arginine (R)), and wherein the amino acid at position 147 or the amino acid at position 213 in the homeostatic domain CH1 of the second chain under b) is independently substituted by glutamine (E) or aspartic acid (D) (according to Kabat EU index numbering).

In one embodiment, the amino acids at positions 124 and 123 in the constitutive domain CL of the second chain are substituted with K (according to the Kabat EU index numbering).

In one embodiment, the amino acid at position 123 in the constant domain CL of the second chain is substituted with R and the amino acid at position 124 is substituted with K (numbering according to the Kabat EU index).

In one embodiment, the amino acids at positions 147 and 213 in the constant domain CH1 of the second light chain are substituted by E (according to the EU index numbering of Kabat).

In one embodiment, the amino acids at positions 124 and 123 in the constant domain CL of the first light chain are substituted with K, and the amino acids at positions 147 and 213 in the constant domain CH1 of the first heavy chain are substituted with E (numbering according to the Kabat EU index).

In one embodiment, the amino acid at position 123 in the constant domain CL of the first light chain is substituted with R and the amino acid at position 124 is substituted with K, and the amino acids at positions 147 and 213 in the constant domain CH1 of the first heavy chain are substituted with E (according to the Kabat EU index numbering).

In one embodiment, the amino acids at positions 124 and 123 in the constant domain CL of the second heavy chain are substituted with K, and wherein the amino acids at positions 147 and 213 in the constant domain CH1 of the second light chain are substituted with E, and the amino acid at position 38 in the variable domain VL of the first light chain is substituted with K, the amino acid at position 39 in the variable domain VH of the first heavy chain is substituted with E, the amino acid at position 38 in the variable domain VL of the second heavy chain is substituted with K, and the amino acid at position 39 in the variable domain VH of the second light chain is substituted with E (numbering according to the Kabat EU index).

In one embodiment, the bispecific antibody is a bivalent antibody comprising a) a first light chain and a first heavy chain of an antibody that specifically binds to a first antigen, and b) a second light chain and a second heavy chain of an antibody that specifically binds to a second antigen, wherein the variable domains VL and VH of the second light chain and the second heavy chain are replaced with each other, and wherein the constant domains CL and CH1 of the second light chain and the second heavy chain are replaced with each other.

The antibody under a) does not contain the modification reported under b), and the heavy chain and light chain under a) are separated chains. In the antibody under b), the variable light chain domain VL in the light chain is replaced by the variable heavy chain domain VH of the antibody, and the constant light chain domain CL is replaced by the constant heavy chain domain CH1 of the antibody; and the variable heavy chain domain VH in the heavy chain is replaced by the variable light chain domain VL of the antibody, and the constant heavy chain domain CH1 is replaced by the constant light chain domain CL of the antibody.

In one embodiment, the bispecific antibody is a bivalent antibody comprising a) a first light chain and a first heavy chain of an antibody that specifically binds to a first antigen, and b) a second light chain and a second heavy chain of an antibody that specifically binds to a second antigen, wherein the constant domains CL and CH1 of the second light chain and the second heavy chain are replaced with each other.

The antibody under a) does not contain the modification reported under b), and the heavy chain and light chain under a) are separated chains. In the antibody under b), the constant light chain domain CL in the light chain is replaced by the constant heavy chain domain CH1 of the antibody, and the constant heavy chain domain CH1 in the heavy chain is replaced by the constant light chain domain CL of the antibody.

In one embodiment, the bispecific antibody is a bispecific antibody comprising a) a full-length antibody that specifically binds to a first antigen and is composed of two antibody heavy chains and two antibody light chains, and b) one, two, three or four single-chain Fab fragments that specifically bind to a second antigen, wherein the single-chain Fab fragment under b) is fused to the full-length antibody under a) at the C-terminus or N-terminus of the heavy chain or light chain of the full-length antibody via a peptide linker.

In one aspect, one or two identical single-chain Fab fragments that bind to a second antigen are fused to the full-length antibody at the C-terminus of the heavy or light chain of the full-length antibody via a peptide linker.

In one aspect, one or two identical single-chain Fab (scFab) fragments that bind to a second antigen are fused to the full-length antibody at the C-terminus of the heavy chain of the full-length antibody via a peptide linker.

In one aspect, one or two identical single-chain Fab (scFab) fragments that bind to a second antigen are fused to the full-length antibody at the C-terminus of the light chain of the full-length antibody via a peptide linker.

In one aspect, two identical single-chain Fab (scFab) fragments that bind to a second antigen are fused to the full-length antibody at the C-terminus of each heavy or light chain of the full-length antibody via a peptide linker.

In one aspect, two identical single-chain Fab (scFab) fragments that bind to a second antigen are fused to the full-length antibody via a peptide linker at the C-terminus of each heavy chain of the full-length antibody.

In one aspect, two identical single-chain Fab (scFab) fragments that bind to a second antigen are fused to the full-length antibody via a peptide linker at the C-terminus of each light chain of the full-length antibody.

In one embodiment, the bispecific antibody is a trivalent antibody comprising a) a full-length antibody that specifically binds to a first antigen and is composed of two antibody heavy chains and two antibody light chains, b) a first polypeptide composed of ba) an antibody heavy chain variable domain (VH), or bb) an antibody heavy chain variable domain (VH) and an antibody constant domain 1 (CH1), wherein the first polypeptide is fused at the N-terminus of its VH domain to the C-terminus of one of the two heavy chains of the full-length antibody via a peptide linker, c) a second polypeptide composed of ca) an antibody light chain variable domain (VL), or cb) an antibody light chain variable domain (VL) and an antibody light chain constant domain (CL), wherein the second polypeptide is fused to the C-terminus of the other of the two heavy chains of the full-length antibody via a peptide linker at the N-terminus of the VL domain, and wherein the antibody heavy chain variable domain (VH) of the first polypeptide and the antibody light chain variable domain (VL) of the second polypeptide together form an antigen binding domain that specifically binds to a second antigen.

In one embodiment, the antibody heavy chain variable domain (VH) of the polypeptide under b) and the antibody light chain variable domain (VL) of the polypeptide under c) are linked and stabilized via an interchain disulfide bridge by introducing a disulfide bond between the following positions: (i) heavy chain variable domain position 44 to light chain variable domain position 100, or (ii) heavy chain variable domain position 105 to light chain variable domain position 43, or (iii) heavy chain variable domain position 101 to light chain variable domain position 100 (always numbered according to the Kabat EU index).

Techniques for introducing non-native disulfide bridges for stabilization are described in, for example, WO 94/029350; Rajagopal, V., et al., Prot. Eng. (1997) 1453-1459; Kobayashi, H., et al., Nucl. Med. Biol. 25 (1998) 387-393; and Schmidt, M., et al., Oncogene 18 (1999) 1711-1721. In one embodiment, the optional disulfide bond between the variable domains of the polypeptides under b) and c) is between heavy chain variable domain position 44 and light chain variable domain position 100. In one embodiment, the optional disulfide bond between the variable domains of the polypeptides under b) and c) is between heavy chain variable domain position 105 and light chain variable domain position 43 (always according to Kabat numbering). In one embodiment, a trivalent bispecific antibody without the optional disulfide bond stabilization between the variable domains VH and VL of the single chain Fab fragment is preferred.

In one embodiment, the bispecific antibody is a trispecific or tetraspecific antibody comprising a) a first light chain and a first heavy chain of a full-length antibody that specifically binds to a first antigen, and b) a second (modified) light chain and a second (modified) heavy chain of a full-length antibody that specifically binds to a second antigen, wherein the variable domains VL and VH are replaced with each other, and/or wherein the constant domains CL and CH1 are replaced with each other, and c) wherein one to four antigen-binding domains that specifically bind to one or two other antigens (i.e., to a third and/or fourth antigen) are fused to the C-terminus or N-terminus of the light chain or heavy chain of a) and/or b) via a peptide linker.

The antibody under a) does not contain the modification reported under b), and the heavy chain and light chain under a) are separate chains.

In one aspect, a trispecific or tetraspecific antibody comprises one or two antigen-binding domains under c) that specifically bind to one or two other antigens.

In one aspect, the antigen binding domain is selected from the group consisting of a scFv fragment and a scFab fragment.

In one aspect, the antigen binding domain is a scFv fragment.

In one aspect, the antigen binding domain is a scFab fragment.

In one aspect, the antigen binding domain is fused to the C-terminus of the heavy chain of a) and/or b).

In one aspect, the trispecific or tetraspecific antibody comprises one or two antigen-binding domains under c) that specifically bind to one other antigen.

In one embodiment, the trispecific or tetraspecific antibody comprises two identical antigen-binding domains under c) that specifically bind to a third antigen. In a preferred embodiment, both of the two identical antigen-binding domains are fused to the C-terminus of the heavy chain of a) and b) via the same peptide linker. In a preferred embodiment, the two identical antigen-binding domains are scFv fragments or scFab fragments.

In one aspect, the trispecific or tetraspecific antibody comprises two antigen-binding domains under c) that specifically bind to the third and fourth antigens. In one embodiment, both of the two antigen-binding domains are fused to the C-terminus of the heavy chain of a) and b) via the same peptide linker. In a preferred embodiment, the two antigen-binding domains are scFv fragments or scFab fragments.

In one embodiment, the bispecific antibody is a bispecific tetravalent antibody comprising a) two light chains and two heavy chains of an antibody that specifically bind to a first antigen (and comprise two Fab fragments), b) two additional Fab fragments of an antibody that specifically bind to a second antigen, wherein both additional Fab fragments are fused to the C-terminus or N-terminus of the heavy chain of a) via a peptide linker, and wherein the following modifications are made in the Fab fragments (i) in the two Fab fragments of a) or in the two Fab fragments of b), the variable domains VL and VH are replaced with each other, and/or the constant domains CL and CH1 are replaced with each other, or (ii) in a) In the two Fab fragments of b), the variable domains VL and VH are replaced with each other, and the constant domains CL and CH1 are replaced with each other, and in the two Fab fragments of b), the variable domains VL and VH are replaced with each other, or the constant domains CL and CH1 are replaced with each other, or (iii) in the two Fab fragments of a), the variable domains VL and VH are replaced with each other, or the constant domains CL and CH1 are replaced with each other, and in the two Fab fragments of b), the variable domains VL and VH are replaced with each other, and the constant domains CL and CH1 are replaced with each other, or (iv) in the two Fab fragments of a), the variable domains VL and VH are replaced with each other, and in the two Fab fragments of b), the constant domains CL and CH1 are replaced with each other. or (v) in the two Fab fragments of a), the constant domains CL and CH1 are replaced with each other, and in the two Fab fragments of b), the variable domains VL and VH are replaced with each other.

In one aspect, the additional Fab fragments are both fused to the C-terminus of the heavy chain of a) or the N-terminus of the heavy chain of a) via a peptide linker.

In one aspect, both of the additional Fab fragments are fused to the C-terminus of the heavy chain of a) via a peptide linker.

In one aspect, both of the additional Fab fragments are fused to the N-terminus of the heavy chain of a) via a peptide linker.

In one embodiment, the following modifications are made in the Fab fragments: in the two Fab fragments in a), or in the two Fab fragments in b), the variable domains VL and VH are replaced with each other, and/or the constant domains CL and CH1 are replaced with each other.

In one embodiment, the bispecific antibody is a tetravalent antibody comprising: a) a (modified) heavy chain of a first antibody that specifically binds to a first antigen and comprises a first VH-CH1 domain pair, wherein the N-terminus of the second VH-CH1 domain pair of the first antibody is fused to the C-terminus of the heavy chain via a peptide linker, b) two light chains of the first antibody of a), c) a (modified) heavy chain of a second antibody that specifically binds to a second antigen and comprises a first VH-CL domain pair, wherein the N-terminus of the second VH-CL domain pair of the second antibody is fused to the C-terminus of the heavy chain via a peptide linker, and d) two (modified) light chains of the second antibody of c), each comprising a CL-CH1 Domain pair.

In one embodiment, the bispecific antibody comprises a) a heavy chain and a light chain of a first full-length antibody that specifically binds to a first antigen, and b) a heavy chain and a light chain of a second full-length antibody that specifically binds to a second antigen, wherein the N-terminus of the heavy chain is linked to the C-terminus of the light chain via a peptide linker.

The antibody under a) does not contain the modification reported under b), and the heavy and light chains are separate chains.

In one embodiment, the bispecific antibody comprises a) a full-length antibody that specifically binds to a first antigen and is composed of two antibody heavy chains and two antibody light chains, and b) an Fv fragment that specifically binds to a second antigen, the fragment comprising a VH2 domain and a VL2 domain, wherein the two domains are linked to each other via a disulfide bridge, wherein only the VH2 domain or the VL2 domain is fused to the heavy chain or light chain of the full-length antibody that specifically binds to the first antigen via a peptide linker.

In the bispecific antibody, the heavy chain and light chain in a) are separate chains.

In one aspect, the other of the VH2 domain or the VL2 domain is fused without a peptide linker to the heavy chain or light chain of a full-length antibody that specifically binds to a first antigen.

In all aspects reported herein, the first light chain comprises a VL domain and a CL domain and the first heavy chain comprises a VH domain, a CH1 domain, a hinge region, a CH2 domain and a CH3 domain.

In one embodiment, the bispecific antibody is a trivalent antibody comprising a) two Fab fragments that specifically bind to a first antigen, b) a CrossFab fragment that specifically binds to a second antigen, wherein the CH1 and CL domains in the fragment are exchanged with each other, c) an Fc region comprising a first Fc region heavy chain and a second Fc region heavy chain, wherein the C-termini of the CH1 domains of the two Fab fragments are linked to the N-termini of the heavy chain Fc region polypeptide, and wherein the C-termini of the CL domain of the CrossFab fragment is linked to the N-terminus of the VH domain of one of the Fab fragments.

In one embodiment, the bispecific antibody is a trivalent antibody comprising a) two Fab fragments that specifically bind to a first antigen, b) a CrossFab fragment that specifically binds to a second antigen, wherein the CH1 and CL domains in the fragment are interchanged, c) an Fc region comprising a first Fc region heavy chain and a second Fc region heavy chain, wherein the C-terminus of the CH1 domain of the first Fab fragment is linked to the N-terminus of one of the heavy chain Fc region polypeptides, and the C-terminus of the CL domain of the CrossFab fragment is linked to the N-terminus of the other heavy chain Fc region polypeptide, and wherein the C-terminus of the CH1 of the second Fab fragment is linked to the N-terminus of the VH domain of the first Fab fragment or the N-terminus of the VH domain of the CrossFab fragment.

In one embodiment, the bispecific antibody comprises a) a full-length antibody that specifically binds to a first antigen and is composed of two antibody heavy chains and two antibody light chains, and b) a Fab fragment that specifically binds to a second antigen, the fragment comprising a VH2 domain and a VL2 domain and comprising a heavy chain fragment and a light chain fragment, wherein the variable light chain domain VL2 in the light chain fragment is replaced by the variable heavy chain domain VH2 of the antibody, and the variable heavy chain domain VH2 in the heavy chain fragment is replaced by the variable light chain and VL2 of the antibody wherein the heavy chain Fab fragment is inserted between the CH1 domain of one of the heavy chains of the full-length antibody and the corresponding Fc region of the full-length antibody, and the light chain Fab The N-terminus of the fragment is bound to the C-terminus of the light chain of the full-length antibody, which is paired with the heavy chain of the full-length antibody and into which the heavy chain Fab fragment has been inserted.

In one aspect, the bispecific antibody comprises a) a full-length antibody that specifically binds to a first antigen and is composed of two antibody heavy chains and two antibody light chains, and b) a Fab fragment that specifically binds to a second antigen, the fragment comprising a VH2 domain and a VL2 domain and comprising a heavy chain fragment and a light chain fragment, wherein the variable light chain domain VL2 in the light chain fragment is replaced by the variable heavy chain domain VH2 of the antibody, and the variable heavy chain domain VH2 in the heavy chain fragment is replaced by the variable light chain domain VL2 of the antibody, and wherein the C-terminus of the heavy chain fragment of the Fab fragment binds to the N-terminus of one of the heavy chains of the full-length antibody, and the C-terminus of the light chain fragment of the Fab fragment The N-terminus of the light chain of the full-length antibody is bound to the N-terminus of the light chain of the full-length antibody, which is paired with the heavy chain of the full-length antibody, and the heavy chain fragment of the Fab fragment is bound to the heavy chain. B. Administration of bispecific antibodies binding to PD-1 and LAG3

For the prevention or treatment of disease, the appropriate dose of the bispecific antibody of the present invention comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3 (used alone or in combination with one or more additional therapeutic agents) will depend on the type of disease to be treated, the route of administration, the weight of the individual, the type of fusion protein, the severity and course of the disease (whether the bispecific antibody is administered for prevention or treatment purposes), previous or concurrent therapeutic interventions, the individual's clinical history and response to the fusion protein, and the judgment of the attending physician. In any case, the practitioner responsible for administration will determine the concentration of one or more active ingredients in the composition and the appropriate dose for a single subject. Various dosing regimens are contemplated herein, including but not limited to single or multiple administrations at various time points, bolus administration, and pulse infusion.

A bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3 as defined herein is suitable for administration to an individual at one time or over a series of treatments. Depending on the type and severity of the disease, about 1 μg/kg to 15 mg/kg (e.g., 0.1 mg/kg – 10 mg/kg) of a bispecific antibody may be an initial candidate dose for administration to an individual, for example, by one or more divided administrations or by continuous infusion. Depending on the factors mentioned above, a typical daily dose may range from about 1 μg/kg to 100 mg/kg or more. For repeated dosing over several days or longer, depending on the condition, treatment will generally continue until a desired suppression of disease symptoms occurs. An exemplary dosage of the bispecific antibody will be in the range of about 0.005 mg/kg to about 10 mg/kg. In other examples, the dosage can also include about 1 μg/kg body weight, about 5 μg/kg body weight, about 10 μg/kg body weight, about 50 μg/kg body weight, about 100 μg/kg body weight, about 200 μg/kg body weight, about 350 μg/kg body weight, about 500 μg/kg body weight, about 1 mg/kg body weight, about 5 mg/kg body weight, about 10 mg/kg body weight, about 50 mg/kg body weight, about 100 mg/kg body weight, about 200 mg/kg body weight, about 350 mg/kg body weight, about 500 mg/kg body weight to about 1000 mg/kg body weight or more per administration and any range derivable therefrom. In an example of a range derived from the numbers listed herein, doses ranging from about 5 mg/kg body weight to about 100 mg/kg body weight, about 5 μg/kg body weight to about 500 mg/kg body weight, etc. can be administered based on the above numbers. Thus, one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 5.0 mg/kg, or 10 mg/kg (or any combination thereof) can be administered to an individual. Such doses can be administered intermittently, such as weekly or every three weeks (e.g., so that the individual receives about two to about twenty, or, for example, about six doses of the fusion protein). An initial higher loading dose can be administered, followed by one or more lower doses. However, other dosage regimens can be used. The progress of this treatment is easily monitored through the use of techniques and measurements.

In one aspect, the bispecific antibody targeting PD-1 and LAG3 is administered to a subject at a fixed dose of about 300 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, or about 1200 mg every three weeks (Q3W), for example, administered at a fixed dose on day 1 (± 1 day) of a three-week dosing cycle.

In a specific aspect, the bispecific antibody targeting PD-1 and LAG3 is administered to a subject at a fixed dose of about 600 mg every three weeks (Q3W), for example, at a fixed dose of 600 mg Q3W (e.g., administered on day 1 (± 1 day) of a three-week dosing cycle). Specifically, the bispecific antibody targeting PD-1 and LAG3 is administered to a subject at a fixed dose of about 600 mg every three weeks (Q3W) on day 1 of a three-week dosing cycle.

In another aspect, the bispecific antibody targeting PD-1 and LAG3 is administered to a subject at a fixed dose of about 1200 mg every three weeks, for example, at a fixed dose of 1200 mg Q3W.

In another aspect, the bispecific antibody targeting PD-1 and LAG3 is administered to the subject at a fixed dose of about 2100 mg every two weeks (Q2W), for example, at a fixed dose of 2100 mg Q2W. C. Administration of Paclitaxel or Albumin-Bound Paclitaxel

Therapeutically effective amounts of a variety of chemotherapeutics, specifically taxanes (e.g., paclitaxel or nab-paclitaxel) are known in the art and are contemplated in the present invention. In a specific instance, paclitaxel is administered intravenously at a dose of 200 mg/m ² once every three weeks. In one instance, the starting dose level of paclitaxel in the study was 200 mg/m ² administered intravenously over 3 hours once every three weeks. In a specific instance, nab-paclitaxel is administered intravenously at a dose of 200-100 mg/m 2 once a week for three weeks, followed by 1 week off. Nab-paclitaxel doses should not be administered more than once every 7 days. In one case, the starting dose level of nab-paclitaxel in the study was 200-100 mg/m2, given intravenously over 3 hours every three 30-minute intervals per week for 3 weeks on a repeat schedule followed by 1 week off. D. Administration of Pemetrexed

Therapeutically effective amounts of a variety of chemotherapeutic agents, particularly anti-metabolic drugs (e.g., pemetrexed) are known in the art and are contemplated in the present invention. In a particular instance, pemetrexed is administered intravenously at a dose of 500 mg/m ² once every three weeks. In one instance, the starting dose level of pemetrexed in the study was 500 mg/m ² administered intravenously once every three weeks. E. Administration of Carboplatin

Therapeutically effective amounts of a variety of chemotherapeutics, particularly platinum-based chemotherapeutics (e.g., carboplatin) are known in the art and are contemplated in the present invention. In a particular instance, carboplatin is administered intravenously at a target area under the concentration-time curve (AUC) of 5 mg/mL·min every three weeks. In one instance, the starting dose level of carboplatin in the study was 100 mg/m ² , which was administered intravenously at a target AUC of 5 mg/mL·min every three weeks. V. Pharmaceutical Compositions and Formulations

Also provided herein are pharmaceutical compositions and formulations comprising bispecific antibodies targeting PD-1 and LAG3 and, optionally, pharmaceutically acceptable carriers. The present disclosure also provides pharmaceutical compositions and formulations comprising bispecific antibodies targeting PD-1 and LAG3 and one or more chemotherapeutic agents (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) albumin-bound paclitaxel), and, optionally, pharmaceutically acceptable carriers. Pharmaceutical compositions and formulations of bispecific antibodies targeting PD-1 and LAG3 and/or other agents described herein (e.g., one or more chemotherapeutic agents, for example, (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) albumin-bound paclitaxel) can be prepared by mixing one or more agents having the desired purity with one or more optionally selected pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. ed. (1980)), in the form of a lyophilized preparation or an aqueous solution.

The pharmaceutical compositions and formulations described herein can be prepared by mixing an active ingredient having a desired purity (e.g., a bispecific antibody targeting PD-1 and LAG3 and/or one or more chemotherapeutic agents (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) albumin-bound paclitaxel)) with one or more optionally selected pharmaceutically acceptable carriers (see, e.g., Remington's Pharmaceutical Sciences 16th edition, Osol, A. ed. (1980)), for example, in the form of a lyophilized preparation or an aqueous solution.

Pharmaceutically acceptable carriers are generally nontoxic to the recipient at the dosages and concentrations employed and include, but are not limited to: buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexadecene diammonium chloride; benzoammonium chloride; benzathonine chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, aspartic acid, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter ions such as sodium; metal complexes (e.g., zinc protein complexes); and/or non-ionic surfactants such as polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers herein further include interstitial drug dispersions, such as soluble neutral active hyaluronidase glycoproteins (sHASEGP), such as human soluble PH-20 hyaluronidase glycoproteins, such as rHuPH20 (HYLENEX®, Baxter International, Inc.). Certain exemplary sHASEGPs and methods of use (including rHuPH20) are described in U.S. Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, sHASEGP is conjugated to one or more additional glycosaminoglycans, such as chondroitinase.

Exemplary lyophilized antibody formulations are described in U.S. Pat. No. 6,267,958. Aqueous antibody formulations include those described in U.S. Pat. No. 6,171,586 and WO2006/044908, the latter formulations including a histidine-acetate buffer.

The formulations described herein may also contain more than one active ingredient suitable for the specific indication being treated, preferably, they are complementary active ingredients that do not adversely affect each other. For example, it may be desirable to further provide an additional therapeutic agent (e.g., a chemotherapeutic agent, a cytotoxic agent, a growth inhibitor and/or an anti-hormonal agent, such as those described herein above). These active ingredients are suitably present in combination in an amount effective for the intended purpose.

The active ingredient can be entrapped in microcapsules (e.g., hydroxymethylcellulose microcapsules or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or macroemulsions, for example, prepared by coacervation techniques or by interfacial polymerization. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. ed. (1980).

Sustained release preparations may be prepared. Suitable examples of sustained release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules.

Formulations intended for in vivo administration are generally sterile. Sterility can be readily achieved, for example, by filtration through a sterile filter membrane. VI. Articles of manufacture or kits A. Kits comprising bispecific antibodies targeting PD-1 and LAG3 and one or more chemotherapeutic agents

In another aspect, provided herein is a product or kit comprising a bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel). In some cases, the article of manufacture or kit further comprises a package insert comprising instructions for use of a chemotherapy agent (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel) in combination with a bispecific antibody targeting PD-1 and LAG3 to treat a cancer (e.g., NSCLC, e.g., squamous or non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV squamous or non-squamous NSCLC) or delay its progression in an individual. Any of the bispecific antibodies targeting PD-1 and LAG3 and/or chemotherapy agents described herein may be included in the article of manufacture or kit.

In another embodiment of the present invention, a kit is provided, comprising a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody being used in combination with a chemotherapeutic agent (e.g., one or both of pemetrexed and carboplatin) for treating an individual having cancer (e.g., non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC, or TNBC, e.g., locally advanced, unresectable or metastatic TNBC) according to any of the methods described herein. In some cases, the kit further comprises a chemotherapeutic agent (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel). In some cases, the product or kit further comprises a package insert comprising instructions for use of a bispecific antibody targeting PD-1 and LAG3 in combination with a chemotherapy agent (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel) to treat or delay the progression of cancer (e.g., non-squamous NSCLC, e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC, or TNBC, e.g., locally advanced, unresectable or metastatic TNBC) in an individual.

In another embodiment of the present invention, a kit is provided, comprising a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody being used in combination with a chemotherapeutic agent (e.g., one or both of paclitaxel and carboplatin) for treating an individual having cancer (e.g., squamous NSCLC, e.g., stage IIIB/IIIC or stage IV non-squamous NSCLC) according to any of the methods described herein. In some cases, the kit further comprises a chemotherapeutic agent (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel). In some cases, the product or kit further comprises a package insert comprising instructions for use of a combination of a bispecific antibody targeting PD-1 and LAG3 and a chemotherapy agent (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel) to treat cancer (e.g., squamous NSCLC, such as stage IIIB/IIIC or stage IV squamous NSCLC) in an individual.

In some cases, the bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapy agents (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) albumin-bound paclitaxel) are in the same container or in separate containers. Suitable containers include, for example, bottles, vials, bags, and syringes. The container can be formed of a variety of materials, such as glass, plastic (such as polyvinyl chloride or polyolefin), or metal alloy (such as stainless steel or hastelloy). In some instances, the container holds the formulation, and a label on or associated with the container can indicate directions for use. The article or kit may further include other materials as desired from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes, and leaflets with instructions for use. In some examples, the article further includes one or more other reagents (e.g., additional chemotherapeutic agents or anti-tumor agents). Suitable containers for one or more agents include, for example, bottles, vials, bags, and syringes.

Any of the bispecific antibodies and/or chemotherapeutics targeting PD-1 and LAG3 described herein (e.g., pemetrexed, paclitaxel, and/or carboplatin) may be included in the article or kit. Any of the articles or kits may include instructions for administering the bispecific antibodies and/or one or more chemotherapeutics targeting PD-1 and LAG3 (e.g., (a) one or both of pemetrexed and carboplatin, or (b) one or both of paclitaxel and carboplatin, or (c) nab-paclitaxel) to a subject according to any of the methods described herein, such as any of the methods described in Section II above. VII. EXAMPLES Example 1 : A Phase II , Randomized, Multicenter, Double-Blind, Controlled Study of RO7247669 Plus Platinum-Based Chemotherapy Versus Pembrolizumab Plus Platinum-Based Chemotherapy in Patients with Previously Untreated Locally Advanced or Metastatic Non-Small Cell Lung Cancer A. Overview of Study Design

CO44194 is a randomized, Phase II, global, multicenter, double-blind study designed to evaluate the efficacy, safety, and pharmacokinetics of RO7247669 in combination with platinum-based chemotherapy compared with pembrolizumab and platinum-based chemotherapy in patients with previously untreated locally advanced unresectable (stage IIIB/IIIC) or metastatic (stage IV) non-small cell lung cancer (NSCLC) who are not eligible for radical surgery and/or definitive chemoradiation.

Approximately 180 patients in the study were randomized in a 1:1 ratio to receive RO7247669 plus platinum chemotherapy (Arm A) or pembrolizumab plus platinum chemotherapy (Arm B) as follows: Arm A (n = approximately 90) : RO7247669 plus platinum chemotherapy. ● Participants with non-squamous (NSQ) NSCLC will receive induction therapy with blinded RO7247669 in combination with pemetrexed and carboplatin, all on Day 1 every 3 weeks (Q3W) for four 21-day cycles, followed by Q3W maintenance therapy with blinded RO7247669 with pemetrexed until disease progression or treatment discontinuation. ● Participants with squamous (SQ) NSCLC will receive blinded RO7247669 in combination with paclitaxel and carboplatin, all on Day 1 Q3W for four 21-day cycles, then blinded RO7247669 (on Day 1) Q3W until disease progression or treatment discontinuation. Arm B (n = approximately 90) : Pembrolizumab plus platinum chemotherapy. ● Participants with NSQ NSCLC will receive induction therapy with blinded pembrolizumab in combination with pemetrexed and carboplatin, all on Day 1 Q3W for four 21-day cycles, followed by maintenance therapy with blinded pembrolizumab in combination with pemetrexed Q3W until disease progression or treatment discontinuation. ● Participants with SQ NSCLC will receive blinded pembrolizumab in combination with paclitaxel and carboplatin, all on Day 1 Q3W for four 21-day cycles, followed by blinded pembrolizumab (on Day 1) Q3W until disease progression or treatment discontinuation.

Treatment continued until disease progression according to RECIST v1.1 occurred. The total duration of study participation for each individual was expected to range from 1 day to a maximum of 58 months.

In this protocol, “study treatment” refers to the combination of treatments assigned to patients as part of the study (i.e., the combination of RO7247669 or pembrolizumab with pemetrexed and carboplatin or paclitaxel and carboplatin).

Several key aspects of the study design and study population are summarized in Table 3. An overview of the studies is presented in Figure 1. Table 3. Summary of the CO44194 study Expect: Phase II Group Type: Adult patients Comparison method: Active comparator Population Diagnosis or Condition: Previously untreated locally advanced unresectable (stage IIIB/IIIC) or metastatic (stage IV) NSCLC who are not eligible for radical surgery and/or definitive chemoradiation. Intervention Mode: Parallel Groups Age of group: 18 years old or above Test compound: RO7247669 Site distribution: Multiple Sites Active comparator: Pembrolizumab Study Intervention Allocation Method: Random grouping and stratification Number of groups: 2 Number of participants to be enrolled: About 180 cases

The objectives and endpoints of the study are presented in Table 4. Table 4. Objectives and Endpoints Main objectives Estimates and Definitions ● To evaluate the efficacy of RO7247669 in combination with platinum-based chemotherapy (Arm A) compared with pembrolizumab plus platinum-based chemotherapy (Arm B). ● Population: Participants with previously untreated locally advanced, unresectable non-small cell lung cancer (NSCLC) who were not eligible for definitive chemoradiation (stage IIIB/IIIC), or metastatic (stage IV) NSCLC with squamous (SQ) or non-squamous (NSQ) histology and no known epidermal growth factor receptor ( EGFR ) or anaplastic lymphoma kinase ( ALK ) genomic tumor aberrations. ● Endpoint: Post-randomization progression-free survival (PFS), defined as the time from randomization to the first occurrence of disease progression or death from any cause, whichever occurred first, as assessed by the investigator according to Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. ● Treatments: – Experimental arms: NSQ NSCLC: RO7247669 in combination with pemetrexed and carboplatin followed by maintenance RO7247669 and pemetrexed. SQ NSCLC: RO7247669 in combination with paclitaxel and carboplatin followed by RO7247669. – Control arms: NSQ NSCLC: pembrolizumab in combination with pemetrexed and carboplatin followed by maintenance pembrolizumab and pemetrexed. SQ NSCLC: pembrolizumab in combination with paclitaxel and carboplatin followed by pembrolizumab. ● Concurrent events and management strategy: – Early discontinuation of study treatment for any reason before the corresponding event of interest: Treatment policy strategy. – Initiation of non-protocol-specified anticancer therapy before the corresponding event of interest: treatment policy strategy. ● Population-level summary: hazard ratio for progression-free survival (PFS). ● To evaluate the efficacy of RO7247669 in combination with platinum-based chemotherapy (Arm A) compared with pembrolizumab plus platinum-based chemotherapy (Arm B). ● Population: As defined above; participants must have measurable disease at baseline as assessed by the investigator according to RECIST v1.1. ● Endpoint: Objective response rate (ORR), defined as the proportion of participants with two consecutive complete responses or partial responses assessed by the investigator according to RECIST v1.1, ≥ 4 weeks apart. ● Treatment: As defined above. ● Concurrent events and management strategies: Definitions. ● Population-level summary: Difference in ORR. Secondary Goals Corresponding endpoint ● To evaluate the efficacy of RO7247669 in combination with platinum-based chemotherapy (Arm A) compared with pembrolizumab plus platinum-based chemotherapy (Arm B). ● Overall survival (OS) after randomization, defined as the time from randomization to death from any cause. ● Duration of response for participants with a confirmed objective response, defined as the time from the first confirmed objective response to disease progression or death from any cause, whichever occurred first, as assessed by the investigator according to RECIST v1.1. ● PFS and OS in participants with programmed death ligand 1 (PD-L1) expression, defined as tumor cells (TC) < 1%, 1%-49%, and ≥ 50%, as assessed by retrospective central PD-L1 testing. ● Changes from baseline to Week 12 in patient-reported outcomes of lung cancer symptoms, physical function, role functioning, and global health status/quality of life, as assessed using the European Organisation for Research and Treatment of Cancer (EORTC) Item Bank. ● To evaluate the safety of RO7247669 combined with platinum-based chemotherapy (Arm A) compared with pembrolizumab plus platinum-based chemotherapy (Arm B). ● The incidence and severity of adverse events, where the severity was determined according to the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events Version 5.0. The severity of cytokine release syndrome was determined according to the American Society of Transplantation and Cellular Therapy consensus grading scale. ● Study the pharmacokinetics (PK) of RO7247669, pemetrexed, carboplatin and paclitaxel. ● PK profile of RO7247669 and derived parameters, including but not limited to the following (as appropriate and when data permit): – Maximum concentration – Duration of maximum concentration – Clearance – Steady-state volume of distribution – Area under the concentration-time curve – Half-life ● Concentration of RO7247669 in serum at specified time points. ● Concentration of calcein (as total platinum), pemetrexed and paclitaxel in plasma at specified time points. ● Assess the immune response to RO7247669. ● The incidence of ADA to RO7247669 at baseline and during the study. Exploratory goals Corresponding endpoint ● To assess the tolerability of RO7247669 plus platinum-based chemotherapy (Arm A) compared with pembrolizumab plus platinum-based chemotherapy (Arm B) from the participants' perspective. ● The presence, frequency, severity, and/or interference with daily functioning of symptomatic treatment toxicity, specified as related to the treatment included in the study (i.e., RO7247669, pembrolizumab, chemotherapy), as assessed using the National Cancer Institute (NCI) Common Criteria for Patient-Reported Outcomes Adverse Events. ● The frequency of participant responses regarding the degree of bothersomeness from treatment symptoms, as assessed using the EORTC Item Library46. ● Assess the potential effects of anti-drug antibodies (ADA). ● Relationship between RO7247669 ADA status and efficacy, safety, or PK endpoints. ● Assess the predictive and prognostic effects of exploratory biomarkers in tissue and blood and their association with disease status, resistance mechanisms, and/or response to the study drug. ● In addition, assess the association of biomarkers with susceptibility to adverse events (i.e., safety biomarkers), evidence of RO7247669 activity (i.e., pharmacodynamic biomarkers), or the potential to increase knowledge and understanding of disease biology and drug safety or pharmacokinetics. ● Relationship between biomarkers in blood and tumor tissues and efficacy, safety, PK or other biomarker endpoints. Blinded RO7247669 and blinded pembrolizumab

Participants received blinded RO7247669 (600 mg every three weeks (Q3W)) or blinded pembrolizumab (200 mg Q3W) by IV infusion. All study treatment administrations were performed in a monitored environment with immediate access to trained personnel and adequate equipment and medications to manage potential severe reactions. The 600-mg dose of RO7247669 Q3W and the 200-mg dose of pembrolizumab Q3W remained constant throughout the study.

The initial dose of RO7247669 and pembrolizumab is delivered by IV infusion over 60 (± 15) minutes. If the 60-minute infusion is tolerated without infusion-related adverse events (fever or chills), the second infusion may be delivered over 30 (± 10) minutes. If the 30-minute infusion is well tolerated, all subsequent infusions may be delivered over 30 (± 10) minutes. Pemetrexed, paclitaxel, and carboplatin

As summarized in Table 5, participants were administered paclitaxel, pemetrexed, and carboplatin. Table 5. Treatment regimen of pemetrexed, paclitaxel, and carboplatin Research treatment Dosage and route of administration Induction period (four 21-day cycles) Maintain (until PD) Pemetrexed 500 mg/m ² by IV infusion. On day 1, Q3W, it took about 10 minutes. On day 1, Q3W, it took about 10 minutes. Paclitaxel 200 mg/m ² by IV infusion. On day 1, Q3W, it took about 3 hours. Not applicable. Card plating AUC 5 mg/mL • min by IV infusion. On day 1, Q3W, it takes about 30-60 minutes. Not applicable. AUC = area under the concentration-time curve; PD = progressive disease; Q3W = every 3 weeks. Paclitaxel

Paclitaxel 200 mg/ ^m2 was administered to participants as an IV infusion Q3W over 3 hours for four cycles per local practice and labeling. All participants were premedicated with oral or IV steroids and antihistamines per approved product labeling and/or standard practice. Additional premedication was performed per standard practice. Paclitaxel was fully administered prior to initiation of the carboplatin dose. Pemetrexed

Pemetrexed 500 mg/ ^m2 was administered to participants as an IV infusion Q3W over 10 minutes until progression or unacceptable toxicity.

All participants were appropriately supplemented with vitamin B12 and folic acid and were on corticosteroid prophylaxis as listed below (or as per local label): ● Folic acid 350-1000 µg oral (PO): Participants must have taken at least five doses of folic acid in the 7 days prior to the first dose of pemetrexed, and folic acid dosing must continue during the full course of therapy and for 21 days after the final dose of pemetrexed. ● Vitamin B12 1000 µg intramuscular (IM) injection: Participants received an IM injection one week prior to the first dose of pemetrexed and every three cycles thereafter. Subsequent vitamin B12 injections may be given on the same day as pemetrexed. ● Antiemetic prophylaxis with dexamethasone 4 mg (or equivalent) PO twice daily: Participants should take dexamethasone the day before, on, and the day after pemetrexed. Higher or additional doses of antiemetic prophylaxis are permitted during Cycles 1 to 4 but should not exceed doses specified by the Transnational Supportive Care Association and the European Society of Medical Oncology.

Area under the calcitonin concentration-time curve (AUC) 5 mg/mL • min: Participants received calcitonin immediately after paclitaxel or methotrexate as an IV infusion over 30-60 minutes, Q3W, for four cycles, as per local practice and labeling. The dose of calcitonin was calculated using the Calvert formula (see below) and should not exceed 750 mg. Calvert formula: Total dose (in milligrams [mg]) = (target AUC) × (creatine clearance [CrCl} + 25). The estimated glomerular filtration rate used in the Calvert formula should not exceed 125 mL/min. Maximum calcitonin dose (mg) = target AUC 5 (mg/mL • min) × (125 + 25) = 5 × 150 mL/min = 750 mg. Benefit - risk assessment

There is a huge unmet need for patients with advanced or metastatic solid tumors. The benefit of cancer immunotherapy using CPI monoclonal antibodies, such as those targeting PD-1/PD-L1, has been observed primarily in a subset of patients with an inflamed tumor phenotype. However, responses are not guaranteed even in this subset of patients due to primary or acquired resistance mechanisms. Upregulated LAG3 expression on T cells is associated with T cell dysfunction, which may lead to adaptive resistance to anti-PD-1/PD-L1 therapy (Sharma et al., Cell, 168: 707-723, 2017). Therefore, blockade of tumor-infiltrating lymphocytes (TILs) by LAG3 may overcome or prevent resistance mechanisms to anti-PD-1/PD-L1 and help restore or increase T cell proliferation and cytotoxic effector functions.

RO7247669 simultaneously targets two major immune checkpoint receptors: PD-1 and LAG3. Such targeting could be used to overcome resistance through co-ligand blockade and subsequent reactivation of TILs, regardless of T regulatory cells, and could potentially delay or prevent the development of LAG3-mediated adaptive resistance mechanisms.

As of the clinical cutoff date of March 1, 2022, RO7247669 was tolerated at doses up to 2100 mg every 2 weeks (Q2W); adverse events (AEs) were manageable, and a consistent safety profile was observed across different solid tumor indications, including NSCLC, and with approved antibodies targeting PD-1.

Currently, as of the data cutoff date of March 1, 2022, relevant efficacy and safety data for RO724766 are available from Study NP41300, a Phase I study in patients with advanced and/or metastatic solid tumors.

In the dose-escalation portion of Study NP41300, the disease control rate (DCR) was 51.4% (18 of 35 evaluable participants) and the objective response rate (ORR) was 17.1% (6 of 35 participants). Responses were observed in CPI-naïve and CPI-experienced participants across multiple tumor types at 600 mg Q2W and 2100 mg Q2W. In the second part of the study, the DCR was 48.3% (28/58) and the ORR was 5.2% (3/58) in patients treated at 2100 mg Q2W. In patients treated with 600 mg Q2W, the DCR was 40% (4/10) and the ORR was 10% (1/10). In patients treated with 600 mg Q3W, the DCR was 28.6% (2/7) and the ORR was 14.3% (1/7).

As of March 01, 2022, a total of 27 NSCLC participants were evaluable for efficacy in study NP41300. Of these, 25 had experienced 2nd/3rd line CPIs, and 2 were CPI-naive but had received other treatments. In this population, the DCR was 51.9%, with 2/27 NSCLC patients experiencing a cPR (ORR 7.4%). The first responder was a 4th line CPI-naive participant treated with 600 mg Q2W. At the time of data cutoff, the patient's response duration was 19.1 months and progression-free survival (PFS) was 22.3 months, and the response is ongoing. The second responder was a participant who experienced a CPI, treated with 600 mg Q2W, with a duration of response (ongoing at the data cutoff date) of 1.9 months. Both participants remain on study treatment.

In Study NP41300, a total of 31 participants with NSCLC were safety evaluable. Most events reported were mild to moderate in severity. Of the 31 patients, 14 experienced Grade 3 AEs. One patient experienced a Grade 4 AE of respiratory disorder. Study drug was interrupted due to this event. This event was ongoing at the data cutoff date. Of the 31 patients, one patient discontinued study treatment due to an AE of Grade 3 stress cardiomyopathy. The patient recovered from this event.

Preliminary clinical results suggest that combination therapy with an anti-PD-1 agent (nivolumab) and an anti-LAG3 agent (relaxalizumab) may provide an incremental benefit compared with anti-PD-1 therapy alone, while having an acceptable safety profile similar to nivolumab monotherapy. In patients with advanced melanoma who had been previously treated with anti-PD-1/PD-L1 therapy, the combination had an objective response rate (ORR) of 11.5% (n = 61), including a disease control rate of 49% (Ascierto et al., Ann Oncol, 28(Suppl 5):mdx440.011, 2017).

Tawbi et al. ( N. Engl. J. Med., 386: 26-34, 2022) report results from the completed phase III trial RELATIVITY-047 (CA224-047; NCT03470922), which evaluated the efficacy and safety of dual LAG3 and PD-1 inhibition therapy with relalizumab (a human IgG4 LAG3 blocker) and nivolumab (a PD-1 blocking antibody) compared with standard of care nivolumab alone. The study enrolled 714 previously untreated patients with histologically confirmed unresectable stage III or IV melanoma. Patients were stratified according to LAG3 expression, PD-L1 expression, BRAF mutation status, and metastatic stage. The primary efficacy endpoint was progression-free survival (PFS) as determined by blinded independent central review (BICR) according to the solid tumor response evaluation criteria version 1.1 (RECIST v1.1). Secondary endpoints included overall survival (OS) and ORR (Tawbi et al., N. Engl. J. Med., 386: 26-34, 2022).

Blinded independent assessment showed that PFS was longer with relaluzumab plus nivolumab than with nivolumab. Patients treated with relaluzumab plus nivolumab had twice the median progression-free survival (PFS) and a 25% reduction in the risk of disease progression or death compared with nivolumab alone (hazard ratio [HR]: 0.75; p = 0.006 by log-rank test). A 12% difference in the landmark PFS between the groups was observed at 12 months. PFS was longer with relaluzumab plus nivolumab compared with single-agent nivolumab, with a slightly higher incidence of adverse events, and health-related quality of life (HRQoL) measures similar to those observed with nivolumab. The combination of relalizumab and nivolumab also demonstrated a PFS advantage over nivolumab in prespecified subgroups. Patients with features typically associated with a worse prognosis, such as visceral metastases, high tumor burden, elevated serum lactate dehydrogenase levels, or mucosal or acral melanoma, had improved outcomes. Expression of LAG3 or PD-L1 did not predict a PFS benefit with the relalizumab plus nivolumab combination versus nivolumab alone. However, a benefit of relalizumab plus nivolumab combination therapy over nivolumab alone was observed in both BRAF mutant and wild-type subgroups (Tawbi et al., N. Engl. J. Med., 386: 26-34, 2022).

In the relalizumab plus nivolumab treatment group, the ORR confirmed by BICR increased numerically from 32.6% (95% confidence interval [CI]: 27.8 to 37.7) with nivolumab alone to 43.1% (95% CI: 37.9 to 48.4). In addition, a nonstatistically significant improvement in OS was also observed with relalizumab plus nivolumab compared with nivolumab alone, corresponding to an HR of 0.80 (95% CI: 27.8 to 37.7). The median OS was not reached (95% CI: 34.2 months to not reached) vs. 34.1 months (95% CI: 25.2 to not reached), respectively.

A total of 470 patients (65.8%) discontinued treatment (237 patients (66.8%) in the relimatumab plus nivolumab group and 233 patients (64.9%) in the nivolumab group), with the majority of discontinuations attributed to disease progression (reported in 36.3% and 46.0% of patients, respectively) (Tawbi et al., N. Engl. J. Med., 386: 26-34, 2022).

Infusion-related adverse reactions occurred in 5.9% of patients receiving the relalizumab plus nivolumab combination and in 3.6% of patients receiving nivolumab. Grade 3 or 4 treatment-related adverse events occurred in 18.9% of patients in the relalizumab plus nivolumab group and in 9.7% of patients in the nivolumab group. The most common grade 3 or 4 treatment-related adverse events in the relalizumab plus nivolumab group included increased lipase levels (in 1.7% of patients), ALT (1.4%) and AST (1.4%), and fatigue (1.1%). Treatment-related adverse events (of any grade) leading to discontinuation occurred in 14.6% of patients in the relalizumab plus nivolumab group and in 6.7% of patients in the nivolumab group. A total of five deaths were reported that were considered to be related to treatment by the investigators: three deaths (0.8%) in the relalizumab plus nivolumab group (hemophagocytic lymphohistiocytosis, acute pulmonary edema, and pneumonia) and two deaths (0.6%) in the nivolumab group (one patient died of sepsis and myocarditis, and one patient died of pneumonia). The most common categories of immune-related adverse events reported in the relalizumab plus nivolumab group were hypothyroidism or thyroiditis (in 18.0% of patients), rash (9.3%), and diarrhea or colitis (6.8%) (Tawbi et al., N. Engl. J. Med., 386: 26-34, 2022).

Given that the optimal mechanisms of immune regulation in the tumor setting are still being explored, rational new approaches are needed to increase response rates to first - line CPI therapy.

The study was divided into three phases : screening, treatment, and follow-up.

Patients were assessed for study eligibility during the 28-day screening period (Days -28 to -1). Patients who were deemed eligible based on the screening assessment were randomized to receive study treatment.

Patients whose tumors had known epidermal growth factor receptor ( EGFR ) mutations or anaplastic lymphoma kinase ( ALK ) gene aberrations were excluded from the study. Patients with non-squamous tumors of unknown EGFR or ALK mutation status required testing prior to enrollment. Patients with squamous tumors of unknown EGFR or ALK mutation status did not require testing before/at screening. Treatment period

Approximately 180 participants were randomly assigned in a 1:1 ratio to treatment groups, with approximately 90 participants enrolled in each group.

Randomization will use permuted block randomization to ensure balanced assignment to treatment groups and will be stratified according to the following criteria: ● PD-L1 expression (tumor proportion score (TPS) or tumor cell (TC) < 1% vs. 1%−49% vs. ≥ 50% by local assay approved by health authorities). ● Histology (SQ NSCLC vs. NSQ NSCLC). ● Smoking history (current/former vs. never smoker).

To adequately represent all PD-L1 expression subgroups (participants with TPS or TC <1%, 1%-49%, and ≥50%) and reflect the natural distribution of PD-L1 expression observed in patients with NSCLC in the first-line treatment setting, the proportion of participants enrolled in the TPS/TC <1% and TPS/TC 1%-49% PD-L1 expression subgroups was capped at approximately 40% of the total planned enrollment (72 participants), as assessed by a PD-L1 immunohistochemistry (IHC) assay approved by local health authorities.

Treatment continued until disease progression according to RECIST v1.1 occurred. The total duration of study participation for each individual was expected to range from 1 day to a maximum of 58 months. Safety Run-in Assessments

An initial safety run-in assessment was performed to assess the safety and tolerability of the new combination of blinded RO7247669 and chemotherapy. A minimum of 12 participants with NSCLC were required for each histology (SQ and NSQ) during the safety run-in assessment. This included a minimum of 6 participants per histology per group, equivalent to approximately 24 participants total. These participants underwent safety assessments after they had the opportunity to complete two treatment cycles (estimated to be approximately 6 months after the first participant was randomized). Participants in each 12-patient group who withdrew from treatment before completing two treatment cycles were also included in the safety run-in assessment. Randomization will continue after the first 12 patients in each histology are enrolled but will be temporarily paused if the upper limit of 22 participants per NSCLC histology is reached before safety lead-in is cleared (approximately 11 participants in arm A and approximately 11 in arm B).

Tumor assessments were performed at baseline and every 6 weeks (± 7 days) for 48 weeks after Day 1 of Cycle 1, regardless of treatment dose delays. After the Week 48 tumor assessment, tumor assessments were required every 9 weeks (± 7 days) thereafter, regardless of dose delays, until radiographic disease progression according to Response Criteria in Solid Tumors, version 1.1 (RECIST v1.1), withdrawal of consent, study discontinuation, or death, whichever occurred first. Participants who remained on treatment after disease progression per RECIST v1.1 had tumor assessments every 6 weeks (± 2 weeks) after the initial documented progression, or more frequently if clinically indicated (regardless of study duration), until treatment was discontinued.

Participants who discontinued treatment for reasons other than radiographic disease progression per RECIST v1.1 (e.g., toxicity, symptom worsening) continued to have scheduled tumor assessments at the same frequency as would have been followed if the participant continued on study treatment thereafter (i.e., every 6 weeks (± 7 days) after Day 1 of Cycle 1 for 48 weeks, then every 9 weeks (± 7 days)) until radiographic progression per RECIST v1.1, withdrawal of consent, study discontinuation, or death, whichever occurred first, regardless of whether the patient started new anticancer therapy.

Participants randomized to the study were asked to complete patient-reported outcome (PRO) questionnaires. Items from the European Organisation for Research and Treatment of Cancer (EORTC) Library (IL) 85, 132, 188, and 17 assess lung cancer symptoms (i.e., fatigue, cough, chest pain, bone pain, and dyspnea), physical and role functioning, and global HRQoL. Selected items from the National Cancer Institute (NCI) Common Terminology Criteria for Patient-Reported Adverse Events (PRO-CTCAE) and EORTC IL46 focus on the presence, frequency, severity, and/or interference with daily functioning and bother associated with treatment with RO7247669, pembrolizumab, and chemotherapy. Ongoing treatment after progression

During the study, participants who met criteria for disease progression (as assessed by the investigator according to RECIST v1.1) and showed evidence of clinical benefit could continue treatment with RO7247669 and pembrolizumab at the investigator's discretion, provided that the participant met all of the following criteria: ● Evidence of clinical benefit. ● Absence of symptoms and signs indicating clear disease progression (including worsening of laboratory values (e.g., new or worsening hypercalcemia)). ● Absence of Eastern Cooperative on Cancer (ECOG) performance status attributable to disease progression. ● Absence of tumor progression at critical anatomical sites that could not be managed with protocol-permitted medical interventions (e.g., meningeal disease).

Investigator assessment of overall tumor response at all time points was based solely on RECIST v1.1.

All participants who discontinued study treatment were followed up until death, loss to follow-up, participant withdrawal, or study termination. Example 2 : Study population

The CO44194 study enrolled approximately 180 previously untreated participants with locally advanced, unresectable SQ or NSQ histologically metastatic NSCLC who were not eligible for radical surgery or definitive chemoradiation and who did not have EGFR mutations or ALK gene aberrations. A. Inclusion Criteria

Potential participants were eligible for inclusion in the study only if all of the following criteria applied: ● Age ≥ 18 years. ● ECOG performance status of 0 or 1. ● Histologically or cytologically documented locally advanced, unresectable (stage IIIB/IIIC) or metastatic (stage IV) NSCLC, not eligible for radical surgery and/or definitive chemoradiation (UICC/AJCC staging system 8th edition). ○ Patients with mixed histological NSCLC and patients with small cell lung cancer were not eligible for inclusion in the study. ● No prior systemic therapy for metastatic NSCLC. ○ Patients who have received prior neoadjuvant, adjuvant chemotherapy, radiation therapy, or chemoradiation therapy with curative intent for non-metastatic disease must have undergone a treatment-free interval of at least 12 months since the last dose of chemotherapy and/or radiation therapy. ● Tumor PD-L1 status by documented local assessment using a health authority-approved PD-L1 IHC assay. ● Measurable disease as defined by RECIST v1.1. ○ Previously irradiated lesions can only be considered measurable disease if disease progression has been clearly documented at that site, as irradiated and previously irradiated lesions are not the only measurable disease. ● Life expectancy ≥ 12 weeks. ● Adequate hematologic and end-organ function as defined by the following laboratory test results within 14 days prior to the start of study treatment (Day 1 of Cycle 1): ○ Absolute neutrophil count (ANC) ≥ 1.5 × 10 ⁹ /L (≥ 1500/µL) without granulocyte colony-stimulating factor, with the following exceptions: Patients with benign ethnic neutropenia (BEN) and an ANC ≥ 1.3 x 10 ⁹ /L (≥ 1300/µL) are eligible. ○ Lymphocyte count ≥ 0.5 x 10 ⁹ /L (500/µL). ○ Platelet count ≥ 100 x 10 ⁹ /L (100,000/µL) without transfusion. ○ Hemoglobin ≥ 90 g/L (≥ 9 g/dL). Patients may receive transfusions or erythropoiesis-stimulating therapy according to local standards of care. ○ Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) ≤ 2.5 × upper limit of normal (ULN), with the following exceptions: patients with documented liver metastases: AST and ALT ≤ 5 × ULN; patients with documented liver or bone metastases: ALP ≤ 5 x ULN. ○ Total bilirubin ≤ 1.5 x ULN, except for the following: Patients with known Gilbert's disease: bilirubin ≤ 3 x ULN. ○ Creatinine clearance (CrCl) ≥ 45 mL/min, calculated using the Cockcroft-Gault formula (Cockcroft and Gault 1976) or determined by a 24-hour urine collection. ○ Albumin ≥ 25 g/L (≥ 2.5 g/dL). ○ For patients not receiving anticoagulation: International Normalized Ratio (INR) and activated partial thromboplastin time (aPTT) ≤ 1.5 × ULN. For patients receiving anticoagulation: stable anticoagulation regimen. ● Negative HIV test at screening. Individuals who were HIV positive at screening were eligible provided they were stable on antiretroviral therapy, had a CD4 count ≥ 200/µL, and had an undetectable viral load. ● Negative for hepatitis B surface antigen at screening. ● Positive hepatitis B surface antibody (HBsAb) test at screening; or negative HBsAb at screening with either a negative hepatitis B core antibody (HBcAb) or a positive total HBcAb test followed by a negative hepatitis B virus (HBV) DNA test (as defined by local laboratories). ● Negative hepatitis C virus (HCV) antibody test at screening; or positive HCV test at screening followed by negative HCV RNA test. ● Adequate cardiovascular function as demonstrated by: ○ New York Heart Association (NYHA) heart failure class II or lower. ○ Baseline corrected QT (by use of Fridericia's formula (QTcF)) interval ≤ 480 ms. If the QTcF interval is longer than 480 ms but shorter than 500 ms, the patient may undergo a cardiac evaluation and, in the absence of clinically significant findings, will be considered for treatment. ○ Resting systolic blood pressure ≤ 150 mmHg and diastolic blood pressure 100 mmHg (average of three or more readings during two or more treatments with brief breaks between each treatment) or no clinically significant hypertension. ○ Resting heart rate between 45 and 100 bpm (or no clinically significant tachycardia). ○ Left ventricular ejection fraction (LVEF) ≥ 50% as assessed by transthoracic echocardiogram (TTE) or multiple gate acquisition (MUGA) scan (TTE preferred test) within 6 months prior to starting study treatment. ● For female participants with childbearing potential: agree to abstain from sex (avoid heterosexual intercourse) or use contraceptive measures, and agree not to donate eggs. ● For male participants: agree to abstain from sex (avoid heterosexual intercourse) or use contraceptive measures, and agree not to donate sperm. B. Exclusion criteria

Potential participants were excluded from the study if any of the following criteria applied to them: ● NSCLC known to have a mutation in the EGFR gene or the ALK fusion oncogene. ○ Patients with non-squamous tumors of unknown EGFR or ALK mutation status need to be tested prior to enrollment (by local or central testing, using a validated health authority-approved test, or in the EU, using a CE-marked test). Patients with squamous tumors of unknown EGFR or ALK mutation status do not need to be tested before/at screening. ● Symptomatic, untreated, or actively progressive central nervous system (CNS) metastases. Asymptomatic patients with CNS lesions may be treated as long as all of the following criteria are met: ● Measurable disease according to RECIST v1.1 must be present outside the CNS. ● Patients have no history of intracranial or spinal hemorrhage. ● Patients have not undergone stereotactic radiotherapy within 7 days before randomization, whole brain radiotherapy within 14 days before randomization, or neurosurgery within 28 days before randomization. ● Patients have no ongoing need for corticosteroids as treatment for CNS disease. ● If patients are receiving anticonvulsant therapy, the dose is considered stable. ● Metastases are limited to the cerebellum or supratentorial regions (i.e., no metastases to the midbrain, pons, medulla oblongata, or spinal cord). ● There was no evidence of interim progression between completion of CNS-directed therapy and initiation of study treatment. ● There was no evidence of significant vasculogenic edema.

Asymptomatic patients with newly detected CNS metastases at screening are eligible to participate in the study after receiving radiation therapy or surgery without repeat screening brain scans. ● Spinal cord compression not definitively treated with surgery and/or radiation, or previously diagnosed and treated spinal cord compression with no evidence of clinical stability for ≥ 2 weeks prior to starting study treatment. ● History of leptomeningeal disease. ● Uncontrolled tumor-related pain. ● Patients requiring analgesia must be on a stable treatment regimen at the start of the study. ○ Symptomatic lesions amenable to palliative radiation therapy (e.g., bone metastases or metastases causing neurologic impact) should be treated prior to starting study treatment. Patients should have recovered from the effects of radiation. There is no minimum recovery period required. ○ Asymptomatic metastatic lesions that may cause functional deficits or intractable pain and further growth (e.g., epidural metastases not currently associated with spinal cord compression) should be considered for locoregional therapy as appropriate prior to enrollment. ● Uncontrolled pleural effusions, pericardial effusions, or ascites requiring repeated drainage (monthly or more frequently). Patients are permitted to use indwelling catheters (e.g., PLEURX ^® ). ● Uncontrolled or symptomatic hypercalcemia (ionized calcium > 1.5 mmol/L, calcium > 12 mg/dL, or corrected calcium greater than ULN). ● Active or history of autoimmune disease or immunodeficiency, including but not limited to myasthenia gravis, myositis, autoimmune hepatitis, systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, antiphospholipid antibody syndrome, Wegener’s granulomatosis, Sjögren’s syndrome, Guillain-Barré syndrome, or multiple sclerosis, with the following exceptions: ○ Patients with a history of autoimmune hypothyroidism who are taking thyroid replacement hormone are eligible to participate in this study. ○ Patients with controlled type 1 diabetes who are receiving insulin therapy are eligible to participate in the study. ○ Patients with eczema, psoriasis, lichen planus simplex, or vitiligo with only dermatologic manifestations (e.g., patients with psoriasis arthritis were excluded) were eligible for the study if the following criteria were met: the rash must cover < 10% of the body surface area; the disease was well controlled at baseline and required only low-potency topical corticosteroids; and there had been no acute exacerbations of underlying disease requiring psoralen plus ultraviolet A radiation, methotrexate, retinol, biologics, oral calcineurin inhibitors, high-potency or oral corticosteroids in the past 12 months. ● History of idiopathic pulmonary fibrosis, organizing pneumonia (e.g., occlusive bronchitis), drug-induced pneumonitis or idiopathic pneumonia, or evidence of active pneumonia on chest computed tomography (CT) scan. History of radiation pneumonitis (fibrosis) with radiation exposure is allowed. ● Active tuberculosis (TB) as documented by a positive purified protein derivative (PPD) skin test or TB blood test and confirmed by a positive chest X-ray within 3 months prior to starting study treatment. Patients with a positive PPD skin test or TB blood test and a subsequent negative chest X-ray may be eligible for the study. ● Untreated latent TB. ● Current treatment with antiviral therapy for HBV or HCV. ● Severe cardiovascular disease within 3 months before randomization, including any of the following: hypertensive crisis or encephalopathy, unstable angina, transient ischemic attack or stroke, congestive heart failure (according to New York Heart Association classification), severe arrhythmia requiring treatment (exceptions are atrial tremor, paroxysmal supraventricular tachycardia), history of thromboembolic events (such as myocardial infarction, stroke or pulmonary embolism), trophic calcification T (TnT) or trophic calcification I (TnI) greater than or equal to the institutional ULN. Patients with TnT or TnI levels between >1 and <2 × ULN are permitted to enroll if the duplicate level is ≤ 1 × ULN. Patients may undergo cardiac evaluation if the duplicate level is between >1 and <2 × ULN and will be considered for treatment in the absence of clinically significant findings. ● Major surgery other than diagnostic within 4 weeks prior to the start of study treatment or anticipated need for major surgery during the study. ● History of malignancy other than NSCLC within 5 years prior to randomization, excluding malignancy with negligible risk of metastasis or death (e.g., 5-year OS rate > 90%) (e.g., adequately treated cervical carcinoma in situ, non-melanoma skin cancer, localized prostate cancer, ductal carcinoma in situ, or stage I uterine cancer). ● Serious infection within 4 weeks prior to the start of study treatment, including but not limited to hospitalization for infection, bacteremia, or severe pneumonia, or any active infection that could affect patient safety. ● Therapeutic oral or IV antibiotic therapy within 2 weeks prior to the start of study treatment. Patients receiving prophylactic antibiotic therapy (e.g., to prevent urinary tract infection or progression of chronic obstructive pulmonary disease) were eligible for the study. ● Previous allogeneic stem cell or solid organ transplant. ● Use of investigational drugs, any other disease, metabolic dysfunction, physical examination findings, or clinical laboratory findings that may affect the interpretation of results or place the patient at high risk for treatment complications are prohibited. ● Treatment with live attenuated vaccines within 4 weeks before the start of study treatment, or anticipation of needing such vaccines during study treatment or within 5 months after the final dose of study treatment. ● Treatment with investigational therapy within 28 days before the start of study treatment. ● Receipt of any anticancer therapy, including hormonal therapy, within 21 days before the start of study treatment. ● Prior treatment with CD137 agonists or immune checkpoint blockade therapy, including but not limited to anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4), anti-T cell immune receptor with Ig and tyrosine-based inhibitory motif domain (TIGIT), anti-PD-1 and anti-PD-L1 therapeutic antibodies, and anti-LAG3 agents. ● Treatment with systemic immune stimulants (including but not limited to interferons and interleukin-2) within 4 weeks or 5 drug elimination half-lives (whichever is longer) prior to the start of study treatment. ● Patients who have been treated with systemic immunosuppressive drugs (including, but not limited to, corticosteroids, cyclophosphamide, azathioprine, methotrexate, thalidomide, and anti-tumor necrosis factor (TNF) agents) within 2 weeks prior to the start of study treatment, or are expected to require systemic immunosuppressive drugs during study treatment, with the following exceptions: ○ Patients who have received acute low-dose systemic immunosuppressive drugs or a one-time pulse dose of systemic immunosuppressive drugs (e.g., 48-hour corticosteroid allergy to contrast agent) are eligible to participate in this study. ○ Patients receiving corticosteroids (e.g., fluocortisol), inhaled or low-dose corticosteroids for chronic obstructive pulmonary disease or asthma, or low-dose corticosteroids for postural hypotension or adrenal insufficiency are eligible for the study. ● History of severe allergic reaction to chimeric or humanized antibodies, fusion proteins, or platinum-containing compounds. ● Known hypersensitivity to Chinese hamster ovary cell products or to any component of RO7247669 or pembrolizumab formulations. ● Known allergy or hypersensitivity or other contraindications to any component of the chemotherapy regimen that the patient may receive during the study. ● Pregnancy or breastfeeding, or anticipated pregnancy during the study, within 4 months after the final dose of RO7247669 and pembrolizumab, or within 6 months after the final dose of paclitaxel, pemetrexed, or carboplatin. ● Known targetable c-ROS oncogene 1 ( ROS1 ), ^BRAFV600E , or rearranged during transfection (RET) proto-oncogene genomic aberrations. Patients with known targetable ROS1 , BRAF ^V600E , or RET genomic aberrations were allowed to enroll in the trial, the only condition being that they were not eligible to receive available targeted therapies. Example 3 : Study Treatment and Concomitant Therapy A. Study Treatment

ERROR! Reference source not found. A description of the specified study treatments used for study CO44194 is provided. Table 6. Study Treatment Descriptions RO7247669 Pembrolizumab Paclitaxel Card plating Pemetrexed use Experiment Comparison Experiment Experiment Experiment Dosage Level 600 mg q3w 200 mg every 3 weeks 200 mg/ ^m2 Q3W AUC 5 mg/mL • min Q3W 500 mg/ ^m2 Q3W Administration IV infusion IV infusion IV infusion IV infusion IV infusion AUC = area under the concentration-time curve; Q3W = every 3 weeks.

In this protocol, “study treatment” refers to the following combinations of treatments assigned to participants in Group A or Group B in this study: Group A : ● For patients with non-squamous (NSQ) NSCLC, induction therapy with blinded RO7247669 in combination with pemetrexed and carboplatin every three weeks (Q3W) for four cycles, followed by maintenance therapy with blinded RO7247669 in combination with pemetrexed Q3W Q3W ● For patients with squamous (SQ) NSCLC, blinded RO7247669 in combination with paclitaxel and carboplatin Q3W for four cycles, followed by blinded treatment with RO7247669 Q3W. Group B : ● For patients with NSQ NSCLC, blinded pembrolizumab in combination with pemetrexed and carboplatin Q3W was used for induction therapy for four cycles, followed by maintenance therapy with blinded pembrolizumab in combination with pemetrexed Q3W. ● For patients with SQ NSCLC, blinded pembrolizumab was used for treatment with paclitaxel and carboplatin Q3W for four cycles, followed by blinded pembrolizumab Q3W. Blinded RO7247669 and blinded pembrolizumab

Participants received blinded RO7247669 (600 mg Q3W) or blinded pembrolizumab (200 mg Q3W) by IV infusion. All study treatment administrations were performed in a monitored environment with immediate access to trained personnel and adequate equipment and medications to manage potential severe reactions. The 600-mg dose of RO7247669 Q3W and the 200-mg dose of pembrolizumab Q3W remained constant throughout the study.

The initial dose of RO7247669 and pembrolizumab is delivered by IV infusion over 60 (± 15) minutes. If the 60-minute infusion is tolerated without infusion-related adverse events (fever or chills), the second infusion may be delivered over 30 (± 10) minutes. If the 30-minute infusion is well tolerated, all subsequent infusions may be delivered over 30 (± 10) minutes.

Participants who experience an infusion-related adverse event may be premedicated with antihistamines and/or antipyretics for subsequent doses at the investigator's discretion and continued beyond the subsequent dose, but the infusion time may not be reduced for that infusion.

RO7247669 and pembrolizumab infusions were administered to participants according to the instructions for use outlined in Error! Reference source not found. Table 7. Administration of the first and subsequent infusions of RO7247669 or pembrolizumab Study Drugs First infusion Subsequent infusion RO7247669 or pembrolizumab infusion ● For the first infusion of RO7247669 and pembrolizumab, no premedication is allowed. ● Vital signs (pulse rate, respiratory rate, blood pressure, and temperature) should be recorded for 60 minutes before the start of the infusion. ● RO7247669 and pembrolizumab are infused over 60 (± 15) minutes. ● If clinically indicated, vital signs should be recorded every 15 (± 5) minutes during the infusion. ● If a patient experienced an infusion-related reaction (IRR) during any prior infusion of RO7247669 or pembrolizumab, premedication with antihistamines and/or antipyretics may be administered for subsequent doses at the investigator’s discretion. ● Vital signs should be recorded for the 60 minutes prior to infusion. ● If the prior infusion was well tolerated without an IRR, RO7247669 and pembrolizumab should be infused over 30 (± 10) minutes, or over 60 (± 15) minutes if the patient experienced an IRR during the prior infusion. ● Vital signs should be recorded during the infusion if clinically indicated. Observation period after RO7247669 or pembrolizumab infusion ● Following infusion of RO7247669 or pembrolizumab, patients begin a 60-minute observation period. ● Vital signs should be recorded at 30 (± 10) minutes after infusion of RO7247669 or pembrolizumab. ● If the patient tolerated the previous RO7247669 or pembrolizumab infusion well without infusion-related adverse events, the observation period after the next and subsequent infusions can be shortened to 30 minutes. ● If the patient experienced an infusion-related adverse event during the previous infusion, the observation period should be 60 minutes. ● If clinically indicated, vital signs should be recorded at 30 (± 10) minutes after the infusion of RO7247669 or pembrolizumab. Paclitaxel

Paclitaxel 200 mg/ ^m2 is administered to participants as an IV infusion Q3W over 3 hours for four cycles per local practice and labeling. All participants should be premedicated with oral or IV steroids and antihistamines per approved product labeling and/or standard practice. Additional premedication should be performed per standard practice. Paclitaxel should be fully administered prior to initiation of the carboplatin dose. Pemetrexed

Pemetrexed 500 mg/ ^m2 was administered to participants as an IV infusion over 10 minutes Q3W until progression or unacceptable toxicity.

All participants were appropriately supplemented with vitamin B12 and folic acid and were on corticosteroid prophylaxis as listed below (or as per local label): ● Folic acid 350-1000 µg oral (PO): Participants must have taken at least five doses of folic acid in the 7 days prior to the first dose of pemetrexed, and folic acid dosing must continue during the full course of therapy and for 21 days after the final dose of pemetrexed. ● Vitamin B12 1000 µg intramuscular (IM) injection: Participants received an IM injection one week prior to the first dose of pemetrexed and every three cycles thereafter. Subsequent vitamin B12 injections may be given on the same day as pemetrexed. ● Antiemetic prophylaxis with dexamethasone 4 mg (or equivalent) PO twice daily: Participants took dexamethasone the day before, on, and the day after pemetrexed. Higher or additional doses of antiemetic prophylaxis were permitted during cycles 1 to 4 but did not exceed doses based on the Multinational Association for Supportive Care (MASCC) and ESMO guidelines (Roila et al. , Ann Oncol, 27: v119-133, 2016).

Calplatin AUC 5 mg/mL • min: Participants receive calplatin as an IV infusion over 30-60 minutes, Q3W, for four cycles immediately following paclitaxel or methotrexate, as per local practice and labeling. The dose of calplatin should be calculated using the Calvert formula (see below) and should not exceed 750 mg. Calvert formula: Total dose (in milligrams (mg)) = (target AUC) × (CrCl + 25). The estimated glomerular filtration rate (GFR) used in the Calvert formula should not exceed 125 mL/min. Maximum calplatin dose (mg) = target AUC 5 (mg/mL • min) × (125 + 25) = 5 × 150 mL/min = 750 mg.

Paclitaxel, pemetrexed, and carboplatin were administered to participants as outlined in Table 8. Treatment regimens for pemetrexed, paclitaxel, and carboplatin Research treatment Dosage and route of administration Induction period (four 21-day cycles) Maintain (until PD) Pemetrexed 500 mg/m ² by IV infusion On the first day, Q3W, it took about 10 minutes On the first day, Q3W, it took about 10 minutes Paclitaxel 200 mg/m ² by IV infusion On day 1, Q3W, about 3 hours Not applicable Card plating AUC 5 mg/mL • min by IV infusion On day 1, Q3W, about 30 to 60 minutes Not applicable AUC = area under the concentration-time curve; PD = progressive disease; Q3W = every 3 weeks. B. Treatments allowed in combination therapy

The following concomitant therapies are contraindicated as described below: ● Oral contraceptives. ● Hormone replacement therapy. ● Palliative radiation therapy (e.g., to treat known bone metastases or relieve pain) as outlined below: ○ For participants without documented progressive radiographic disease, maximal support for symptom management and avoidance of radiation therapy that would interfere with assessment of tumor target lesions are strongly recommended. Treatment with RO7247669 and pembrolizumab may be continued during palliative radiation therapy. ● Prophylactic or therapeutic anticoagulation (e.g., stable-dose warfarin or low molecular weight heparin). ● Vaccinations (e.g., influenza, COVID-19). Live attenuated vaccines are not permitted. ● Meprogesterone acetate given as an appetite stimulant. ● Mineral corticosteroids (e.g., fluocortisol). ● Inhaled or low-dose corticosteroids for chronic obstructive pulmonary disease or asthma. ● Low-dose mineralocorticoids for orthostatic hypotension or low-dose mineralocorticoids and corticosteroids for adrenocortical insufficiency.

Premedication with pemetrexed, paclitaxel, and carboplatin was permitted. Premedication with antihistamines, antipyretics, and/or analgesics was permitted only for the second and subsequent infusions of RO7247669 and pembrolizumab at the investigator's discretion.

In general, investigators should manage the care of participants in accordance with local standard practices with supportive therapies other than those they define as cautious or contraindicated as clinically indicated. Participants who experience infusion-related symptoms may be treated symptomatically with acetaminophen, ibuprofen, diphenhydramine, and/or _H2 -receptor antagonists (e.g., famotidine, cimetidine), or equivalent medications in accordance with local standard practices. Severe infusion-related events manifested by dyspnea, hypotension, wheezing, bronchospasm, tachycardia, decreased oxygen saturation, or respiratory distress should be treated with supportive care (eg, supplemental oxygen and beta ₂ -adrenaline agonists) as clinically indicated.

Unless the investigator determines that prescription and over-the-counter medications (including vitamins and dietary or herbal supplements) will not interfere with the study, participants must abstain from the medication for 7 days (or 14 days if the medication is a potential enzyme inducer) or 5 drug elimination half-lives (whichever is longer) before starting study treatment and until the completion of the follow- up visit.

Systemic corticosteroids, immunosuppressive drugs, and TNF inhibitors may attenuate the potential beneficial immunologic effects of treatment with RO7247669 and/or pembrolizumab. Therefore, alternative agents, including antihistamines, should be considered in the setting of conventional administration of systemic corticosteroids, immunosuppressive drugs, or TNF inhibitors. If alternatives are not feasible, systemic corticosteroids, immunosuppressive drugs, and TNF inhibitors may be administered at the investigator's discretion.

Systemic corticosteroids or immunosuppressive drugs are recommended, at the discretion of the investigator, for specific adverse events associated with treatment with RO7247669 and pembrolizumab.

Concurrent use of herbal therapies is not recommended because their pharmacokinetics, safety, and potential for drug interactions are generally unknown. However, during the course of a study, the investigator may, at their discretion, use herbal therapies that are not intended to treat cancer.

As described below, the following concomitant therapies are prohibited: ● Investigational therapies (other than study treatments specified in the protocol) within 28 days prior to and during study treatment. ● Concomitant therapies intended to treat cancer (including but not limited to chemotherapy, hormonal therapy, immunotherapy, radiation therapy, and herbal therapies), whether approved by health authorities or experimental (except palliative radiation therapy, radiation therapy to the brain, and local therapies in certain circumstances), at various times prior to and during study treatment (depending on the agent) and until disease progression is documented and the participant stops study treatment. ● Live attenuated vaccines within 4 weeks prior to starting study treatment, during study treatment, and within 5 months after the final dose of study treatment. ● Systemic immune stimulants (including but not limited to interferons and interleukin-2) within 4 weeks prior to starting study treatment or within 5 drug elimination half-lives (whichever is longer) and during study treatment, because these agents may increase the risk of autoimmune conditions when given in combination with study treatment. Example 4 : Efficacy and Safety Assessment A. Efficacy Assessment Tumor Assessment

Participants in the CO44194 study had tumor assessments at screening, every 6 weeks (± 7 days) for the first 48 weeks after the start of treatment (Day 1 of Cycle 1), and every 9 weeks (± 7 days) thereafter, regardless of dose delays. Participants continued to have tumor assessments until radiographic disease progression according to RECIST v1.1 or loss of clinical benefit (for participants continuing on treatment after radiographic progression) (as determined by the investigator), withdrawal of consent, death, or study discontinuation, whichever occurred.

Participants who remained on treatment after investigator-assessed disease progression per RECIST v1.1 had tumor assessments every 6 weeks (± 2 weeks) after the initial documented progression, or more frequently if clinically indicated (regardless of study duration), until treatment was discontinued. Tumor assessments could be repeated at any time if progressive disease was suspected, at the investigator's discretion. Participants who discontinued study treatment in the absence of investigator-assessed radiographic disease progression per RECIST v1.1 (for any reason, including, but not limited to, clinical decline or toxicity) continued to have tumor response assessments at the same frequency until radiographic disease progression per RECIST v1.1, withdrawal of consent, death, or study termination, whichever occurred first. Tumor assessments continued in the absence of radiographic disease progression per RECIST v1.1 regardless of whether participants started new anticancer therapy.

All measurable and/or evaluable lesions were assessed and recorded at Screening. Tumor assessments performed as standard of care prior to obtaining informed consent and within 28 days prior to initiating study treatment do not need to be repeated at Screening as long as they meet the criteria outlined herein.

Objective responses were assessed by the investigator at designated time points according to RECIST v1.1. Whenever possible, assessments were performed by the same individual to ensure internal consistency across visits.

Other endpoints (e.g., PFS, OS, DOR) were calculated programmatically based on investigator response assessments at each specified time point.

The PRO questionnaire was completed to assess the therapeutic benefit of RO7247669 plus platinum chemotherapy. In addition, the PRO questionnaire was able to capture each participant's direct experience of RO7247669 plus platinum chemotherapy. PRO data were collected using the following questionnaires: EORTC IL85, IL132, IL188, and IL17.

The IL85 consists of five lung cancer-specific items from the EORTC that assess cough, shortness of breath, and chest pain. The IL85 takes approximately 3 minutes to complete.

The IL132 consists of three items from the EORTC that are relevant to patients with cancer and are used to assess fatigue. The IL132 takes approximately 2 minutes to complete.

IL188 is a single item from the EORTC used to assess bone pain in patients with cancer. IL188 takes approximately 1 minute to complete.

The IL17 consists of nine items from the EORTC relevant to patients with cancer that assess physical functioning, role functioning, and GHS/QoL. The IL17 takes approximately 4 minutes to complete.

The recall period for these IL assessments was specified as the past week. B. Safety Assessment

Patient-reported outcome (PRO) questionnaires were completed to assess the treatment effects of RO7247669 plus platinum chemotherapy and pembrolizumab plus platinum chemotherapy. In addition, the PRO questionnaire was able to capture each participant's direct experience with the combination of RO7247669 and platinum chemotherapy.

PRO data were collected using the following questionnaires: items selected from NCI PRO-CTCAE and EORTC IL46.

The NCI PRO-CTCAE is a validated item bank that characterizes the presence, frequency, severity, and/or interference with daily functioning of 78 patient-reportable symptomatic treatment toxicities (Basch et al., J Natl Cancer Inst, 106: 1-11, 2014; Dueck et al., JAMA Oncol, 1: 1051-1059, 2015). The NCI PRO-CTCAE consists of 124 questions that are rated on a dichotomous (presence or absence) or 5-point Likert scale (frequency, severity, and interference with daily functioning). Treatment toxicities may occur with observable signs (e.g., vomiting) or non-observable symptoms (e.g., nausea). The standard recall period for the NCI PRO-CTCAE is the previous 7 days.

A subset of eight signs and symptoms that are considered best amenable to current treatments were selected for this study. These signs and symptoms were chosen based on known side effects of the commercially available drugs included in standard of care (i.e., pembrolizumab and platinum-based chemotherapy).

The EORTC IL46 is a validated single-item questionnaire used to assess the overall impact of side effects. The standard EORTC IL46 recall period is the previous week. Example 5 : Statistical Considerations Statistical Assumptions

The purpose of this study was to generate hypotheses regarding the efficacy of RO7247669 plus platinum-based chemotherapy (Arm A) versus pembrolizumab plus platinum-based chemotherapy (Arm B) based on the primary endpoints, investigator-assessed objective response rate (ORR) and progression-free survival (PFS). No formal statistical hypotheses were tested for this study. Sample size determination

A total sample size of approximately 180 patients was planned for this study.

With this sample size, the 95% CI for a 20% improvement in ORR in Arm A relative to Arm B (assuming an ORR of 48% in Arm B) (ie, ΔORR) would be 3.7% to 34.9%. The primary analysis of ORR was performed approximately 3 months after the last patient was randomized.

The primary analysis of PFS was performed when approximately 136 total PFS events had been observed. This is expected to occur approximately 37 months after the first patient was randomized. Assuming an exponential distribution of event times, the 95% confidence interval (CI) would be 0.50 to 0.98 for a target PFS HR of 0.7. ERROR! Reference source not found. and 10 CIs showing several possible true potential improvements in ORR and PFS favoring Arm A. Table 9. Confidence intervals for several possible true potential ΔORR values True potential ΔORR 10% 15% 20% 95% CI ^a -6.4, 25.7 -1.4, 30.3 3.7, 34.9 ^aCalculated using Newcombe's method. Table 10. Confidence intervals for several possible true potential PFS HR values True potential PFS hazard ratio (HR) 0.75 0.7 0.65 95% CI 0.54, 1.05 0.50, 0.98 0.46, 0.91 Analysis Set

The participant analysis set used for analysis purposes is defined in Error! Reference source not found. Table 11. Participant Analysis Set Participant Analysis Set describe Full analysis set All randomly assigned participants. Participants were included in the analysis according to the intervention they were randomly assigned to receive. Measurable at baseline All randomized participants had measurable disease at baseline, as assessed by the investigator according to RECIST v1.1. Participants were included in the analysis according to the intervention they were randomly assigned to receive. Security Analysis Set All participants were exposed to the study intervention; participants were analyzed according to the intervention they actually received. If a participant received any amount of RO7247669 or pembrolizumab, respectively, regardless of initial treatment assignment at randomization, that participant was included in Arm A (RO7247669 + chemotherapy) or Arm B (pembrolizumab + chemotherapy) in the safety analysis. Statistical analysis

Unless otherwise specified, all efficacy analyses were performed on the full analysis set. ORR analyses were performed among all randomized participants with measurable disease at baseline, as determined by the investigator according to RECIST v1.1. No formal hypothesis testing was performed; any treatment group comparisons and their associated p-values were generated for descriptive purposes only.

Unless otherwise specified, all safety analyses were performed on the safety-evaluable population.

The co-primary efficacy endpoints were confirmed ORR and PFS as assessed by the investigator according to RECIST v1.1.

The analysis population for ORR was all randomized patients with measurable disease at baseline. The primary analysis of the primary endpoint of investigator-assessed ORR was performed once all randomized patients had been treated and followed until they had a post-baseline tumor assessment 12 weeks or 3 months after the last patient was randomized, whichever occurred first.

Investigator-assessed PFS was analyzed in all randomized patients. An interim analysis of investigator-assessed PFS was performed at the time of the primary analysis of ORR. The primary analysis of the primary endpoint of investigator-assessed PFS occurred after approximately 136 events had been observed.

ORR was defined as the percentage of participants who experienced two consecutive complete responses or partial responses ≥4 weeks apart, as assessed by the investigator according to RECIST v1.1. Participants who did not undergo a post-baseline overall response assessment were counted as non-responders.

Estimates of the difference in ORR between the two groups and their 95% confidence intervals (CIs) were calculated using the Newcombe method. The 95% CI for the confirmed ORR for each treatment group was calculated using the Wilson score method. The Cochran-Mantel-Haenszel test was used to compare the ORR between the two treatment groups, stratified according to protocol-defined stratification factors.

PFS is defined as the time from randomization to the date of first documented disease progression or death, whichever occurs first. Disease progression for PFS analysis is based on investigator assessment using RECIST v1.1. Data from participants who do not have disease progression or have died at the time of analysis will be censored at the last tumor assessment. Data from participants who do not have a post-baseline tumor assessment will be censored at the randomization date.

A stratified Cox proportional hazards model was used to estimate the HR and its 95% CI. A two-sided stratified log-rank test was used to compare PFS between the two treatment groups. The PFS curve and median PFS for each treatment group were estimated using the Kaplan-Meier method. The Brookmeyer-Crowley method was used to construct the 95% CI for the median PFS for each treatment group. Example 6 : A phase II , multicenter, randomized, double-blind study of RO7247669 in combination with nab-paclitaxel versus pembrolizumab in combination with nab-paclitaxel in participants with previously untreated PD-L1- positive, locally advanced unresectable or metastatic triple-negative breast cancer C. Overview of study design

CO44194 is a phase II, randomized, double-blind, global, multicenter study designed to evaluate the efficacy, safety, and pharmacokinetics of the combination of RO7247669 and nab-paclitaxel compared with pembrolizumab plus nab-paclitaxel in patients with previously untreated locally advanced, unresectable or metastatic (stage IV) PD-L1-positive triple-negative breast cancer (TNBC). The design of the study is shown in Figures 1 and 2A-2C.

Participants were selected based on positive PD-L1 expression on tumors, as assessed by central testing, defined as meeting at least one of the following thresholds specific to each assay: Investigational Dako PD-L1 immunohistochemistry (IHC) 22C3 pharmDx assay Composite Positivity Score (CPS) ≥ 10 and/or Investigational VENTANA PD-L1 (SP263) assay Tumor Area Positivity Score (TAP) ≥ 5% and/or Investigational VENTANA PD-L1 (SP142) assay tumor infiltrating immune cells (ICs) expressing PD-L1 ≥ 1%. CPS is the number of PD-L1-staining cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells, multiplied by 100. IC is defined as the presence of discernible PD-L1 staining of any intensity in tumor-infiltrating immune cells covering the tumor area occupied by tumor cells, associated intratumoral stroma, and continuous peritumoral stroma. TAP is defined as the percentage of stained tumor and immune cells within the total tumor area.

The primary population of interest was all randomized participants, hereafter referred to as the full analysis set (FAS). The 22C3-positive analysis set was defined as all randomized participants with a CPS ≥ 10 using the investigational Dako PD-L1 IHC 22C3 pharmDx assay. The SP263-positive analysis set was defined as all randomized participants with a TAP ≥ 5% using the investigational VENTANA PD-L1 (SP263) assay. The SP142-positive analysis set was defined as all randomized participants with an IC ≥ 1% using the investigational VENTANA PD-L1 (SP142) assay. The safety analysis set (SAS) was defined as all randomized participants who received at least one dose of any study treatment.

The objectives and corresponding endpoints of the study are provided in Table 12 below. Progression-free survival (PFS) is the primary endpoint. Table 12. Objectives and Endpoints Main objectives The corresponding end point ● To evaluate the efficacy of RO7247669 plus nab-paclitaxel compared with pembrolizumab plus nab-paclitaxel in the full analysis set (FAS). ● Progression-free survival (PFS), defined as the time from randomization to the first occurrence of disease progression (as assessed by the investigator according to Response Evaluation Criteria in Solid Tumors (RECIST v1.1)) or death from any cause, whichever occurred first. Secondary Goals Corresponding endpoint ● To evaluate the efficacy of RO7247669 plus nab-paclitaxel compared with pembrolizumab plus nab-paclitaxel in FAS. ● Objective response rate (ORR), defined as the proportion of participants with two consecutive complete responses (CR) or partial responses (PR) of ≥ 4 weeks, as assessed by the investigator according to RECIST v1.1. ● Duration of response (DOR), defined as the time from the first confirmed objective response to the first occurrence of disease progression (as assessed by the investigator according to RECIST v1.1) or death from any cause, whichever occurred first. ● Overall survival (OS), defined as the time from randomization to death from any cause. ● PFS rate at 12 months, defined as the proportion of participants who did not experience disease progression or death from any cause 12 months after randomization, as assessed by the investigator according to RECIST v1.1. ● OS rate at 12 months, defined as the proportion of participants who had not experienced death from any cause 12 months after randomization. ● To evaluate the efficacy of RO7247669 plus nab-paclitaxel compared with pembrolizumab plus nab-paclitaxel in the SP263-positive analysis set, the 22C3-positive analysis set, and the SP142-positive analysis set. ● PFS. ● ORR. ● OS. ● To assess the safety of RO7247669 plus nab-paclitaxel compared with pembrolizumab plus nab-paclitaxel in the safety analysis set (SAS). ● The incidence and severity of adverse events, with severity determined based on the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE) v5.0. ● Changes from baseline in targeted clinical laboratory test results. ● Changes from baseline in targeted vital signs. ● Characterize the pharmacokinetic (PK) profile of RO7247669. ● The PK profile and parameters derived for RO7247669 include, but are not limited to (where appropriate and data permitting) the following parameters: – Maximum concentration – Duration of maximum concentration – Clearance – Steady-state volume of distribution – Area under the concentration-time curve – Half-life. ● Evaluate the immunogenicity of RO7247669. ● The incidence and titer of RO7247669 anti-drug antibodies (ADA) during the study period relative to baseline ADA incidence, and the impact of ADA on exposure, activity, and safety ● Exploratory goals ● Corresponding endpoint ● Evaluate the potential relationship between drug exposure and the efficacy and safety of RO7247669 ● The relationship between serum concentrations or PK parameters of RO7247669 and efficacy endpoints. ● The relationship between serum concentrations or PK parameters of RO7247669 and safety endpoints. ● Assess the potential PK interaction between RO7247669 and nab-paclitaxel ● Based on historical data, RO7247669 PK parameters when RO7247669 is co-administered with nab-paclitaxel are compared to RO7247669 given alone. ● Based on historical data, paclitaxel concentrations when RO7247669 is co-administered with nab-paclitaxel are compared to nab-paclitaxel given alone. ● Identify and/or assess biomarkers that predict response to RO7247669 plus nab-paclitaxel (i.e., predictive biomarkers), are early surrogate markers of efficacy, are associated with progression to a more severe disease state (i.e., prognostic biomarkers), are associated with acquired resistance to RO7247669 plus nab-paclitaxel, are associated with susceptibility to developing adverse events or may lead to improved adverse event monitoring or studies (i.e., safety biomarkers), may provide evidence of activity of RO7247669 plus nab-paclitaxel (i.e., pharmacodynamic biomarkers), or may increase knowledge and understanding of disease biology and drug safety or pharmacokinetics. ● The relationship between biomarkers in blood and tumor tissue can be correlated with efficacy, safety, PK, immunogenicity or other biomarker endpoints.

Some key aspects of the study design and study population are summarized in Table 13 below. Table 13. Overall design and study population Expect: Phase II Group Type: Adult patients Comparison method: Active comparator Population Diagnosis or Condition: Locally advanced, unresectable or metastatic (stage IV) PD-L1 positive triple-negative breast cancer Intervention Mode: Parallel Groups Age of group: ≥ 18 years old Test compound: RO7247669 Site distribution: Multiple sites and multiple regions Active comparator: Pembrolizumab Study Intervention Allocation Method: Random grouping and stratification Number of groups: 2 Number of participants to be enrolled: About 160 cases Research treatment

Approximately 160 participants were randomized in the study to receive RO7247669 plus nab-paclitaxel (Arm A) or pembrolizumab plus nab-paclitaxel (Arm B) as follows: 1. Arm A (n = approximately 80): RO7247669 plus nab-paclitaxel – Participants received blinded RO7247669, administered every 3 weeks (a “cycle”), plus nab-paclitaxel, administered weekly, on a 3-week schedule followed by 1 week off until disease progression or treatment discontinuation. 2. Group B (n = approximately 80 patients): Pembrolizumab plus nab-paclitaxel – Participants received blinded pembrolizumab every 3 weeks (one “cycle”) plus nab-paclitaxel every week for 3 weeks followed by 1 week off until disease progression or treatment discontinuation. Blinded RO7247669 and blinded pembrolizumab

Administration of blinded RO7247669 (600 mg every three weeks (Q3W)) or blinded pembrolizumab (200 mg Q3W) by IV infusion was performed in a monitored environment with immediate access to trained personnel and adequate equipment and medications to manage potentially severe reactions.

The initial dose is delivered over 60 (± 15) minutes. If the 60-minute infusion is tolerated without infusion-related adverse events (fever or chills), the second infusion may be delivered over 30 (± 10) minutes. If the 30-minute infusion is well tolerated, all subsequent infusions may be delivered over 30 (± 10) minutes.

Participants who experience an infusion-related adverse event may premedicate with antihistamines and/or antipyretics for the next infusion, but the infusion time for that infusion may not be reduced. Nab- paclitaxel

The starting dose level of nab-paclitaxel in this study was 100 mg/m ² , given intravenously over 30 minutes weekly, on a repeat schedule of 3 weeks, followed by a 1-week break. The dose of nab-paclitaxel should not be administered more frequently than once every 7 days.

On days when an infusion of blinded RO7247669 or pembrolizumab and nab-paclitaxel was planned, chemotherapy was administered after the infusion of RO7247669 or pembrolizumab.

Dosage modifications of RO7247669 or pembrolizumab are not permitted. Dose modifications of nab-paclitaxel may be implemented based on local guidelines and practices. Participation Period

Treatment continued until investigator-assessed disease progression per the Solid Tumor Response Evaluation Criteria v1.1 occurred. The total duration of study participation for each individual was expected to range from 1 day to more than 30 months. Benefit - Risk Assessment

The study aims to evaluate the efficacy, safety, and pharmacokinetics of RO7247669, a novel immunomodulatory therapy, in combination with chemotherapy to address a significant unmet medical need in patients with previously untreated, locally advanced, unresectable or metastatic PD-L1-positive TNBC who are not eligible for curative surgery and/or definitive chemoradiation.

There is a huge unmet need for patients with advanced or metastatic solid tumors. The benefit of cancer immunotherapy using CPI monoclonal antibodies, such as those targeting PD-1/PD-L1, has been observed primarily in a subset of patients with an inflamed tumor phenotype. However, responses are not guaranteed even in this subset of patients due to primary or acquired resistance mechanisms. Nearly all patients with metastatic TNBC eventually progress and die from the disease, so even incremental progress in therapy is critical. Upregulated LAG3 expression on T cells is associated with T cell dysfunction, which may lead to adaptive resistance to anti-PD-1/PD-L1 therapy (Sharma et al., Cell , 168: 707-723, 2017). Therefore, blockade of tumor-infiltrating lymphocytes (TILs) by LAG3 may overcome or prevent resistance mechanisms to anti-PD-1/PD-L1 and help restore or increase T cell proliferation and cytotoxic effector function.

RO7247669 simultaneously targets two major immune checkpoint receptors: PD-1 and LAG3. Such targeting could be used to overcome resistance through co-ligand blockade and subsequent reactivation of TILs, regardless of T regulatory cells, and could potentially delay or prevent the development of LAG3-mediated adaptive resistance mechanisms.

Indeed, preliminary clinical results suggest that combination therapy with an anti-PD-1 agent (nivolumab) and an anti-LAG3 agent (relaxalizumab) may provide an incremental benefit compared with anti-PD-1 therapy alone, while having an acceptable safety profile similar to nivolumab monotherapy. In patients with advanced melanoma previously treated with anti-PD-1/PD-L1 therapy, the combination resulted in an objective response rate (ORR) of 11.5% (n = 61), with a disease control rate (DCR) of 49% (Ascierto et al., Ann Oncol, 28(Suppl 5):mdx440.011, 2017).

Tawbi et al. ( N Eng J Med , 386: 26-34, 2022) reported results from the completed phase III trial RELATIVITY-047 (CA224-047; NCT03470922), which evaluated the efficacy and safety of dual LAG3 and PD-1 inhibition therapy with relalizumab (a human IgG4 LAG3-blocking antibody) and nivolumab (a PD-1-blocking antibody) compared with standard of care nivolumab alone. The study enrolled 714 previously untreated patients with histologically confirmed unresectable stage III or IV melanoma. Patients were stratified according to LAG3 expression, PD-L1 expression, BRAF mutation status, and metastatic stage. The primary efficacy endpoint was PFS as assessed by blinded independent central review (BICR) according to RECIST v1.1. Secondary endpoints included OS and ORR.

Blinded independent assessment showed that PFS was longer with relaluzumab plus nivolumab than with nivolumab. Patients treated with relaluzumab plus nivolumab had twice the median progression-free survival (PFS) and a 25% reduction in the risk of disease progression or death compared with nivolumab alone (hazard ratio [HR]: 0.75; p = 0.006 by log-rank test). A 12% difference in the landmark PFS between the groups was observed at 12 months. PFS was longer with relaluzumab plus nivolumab compared with single-agent nivolumab, with a slightly higher incidence of adverse events, and health-related quality of life (HRQoL) measures similar to those observed with nivolumab. The combination of relaluzumab and nivolumab also demonstrated a PFS advantage over nivolumab in prespecified subgroups. Patients with features typically associated with a worse prognosis, such as visceral metastases, high tumor burden, elevated serum lactate dehydrogenase levels, or mucosal or acral melanoma, had improved outcomes. Expression of LAG3 or PD-L1 did not predict a PFS benefit with the relaluzumab plus nivolumab combination versus nivolumab. However, a benefit with the relaluzumab plus nivolumab combination over nivolumab was observed in both the BRAF mutant and wild-type subgroups (Tawbi et al., N Eng J Med , 386: 26-34, 2022).

In the relalizumab plus nivolumab treatment group, the ORR confirmed by BICR increased numerically from 32.6% (95% CI: 27.8 to 37.7) with nivolumab alone to 43.1% (95% CI: 37.9 to 48.4). In addition, a non-statistically significant improvement in OS was also observed with relalizumab plus nivolumab compared with nivolumab alone, corresponding to an HR of 0.80 (95% CI: 27.8 to 37.7). Median OS was not reached (95% CI: 34.2 months to not reached) vs. 34.1 months (95% CI: 25.2 to not reached) in the relalizumab plus nivolumab group and in the nivolumab alone group, respectively.

A total of 470 patients (65.8%) discontinued treatment (237 patients (66.8%) in the relimatumab plus nivolumab group and 233 patients (64.9%) in the nivolumab group), with the majority of discontinuations attributed to disease progression (reported in 36.3% and 46.0% of patients, respectively) (Tawbi et al., N. Engl. J. Med , 386: 26-34, 2022).

Infusion-related adverse reactions occurred in 5.9% of patients receiving the relalizumab plus nivolumab combination and in 3.6% of patients receiving nivolumab alone. Grade 3 or 4 treatment-related adverse events occurred in 18.9% of patients in the relalizumab plus nivolumab group and in 9.7% of patients in the nivolumab group. The most common grade 3 or 4 treatment-related adverse events in the relalizumab plus nivolumab group included increased lipase levels (reported in 1.7% of patients), alanine transaminase (ALT) (1.4%) and aspartate transaminase (AST) (1.4%), and fatigue (1.1%). In the relalizumab-plus-nivolumab group, 14.6% of patients experienced treatment-related adverse events (of any grade) leading to discontinuation and in 6.7% of patients in the nivolumab group. A total of five treatment-related deaths were reported and considered by the investigators to be treatment-related: three deaths (0.8%) in the relalizumab-plus-nivolumab group (hemophagocytic lymphohistiocytosis, acute pulmonary edema, and pneumonia) and two deaths (0.6%) in the nivolumab group (one patient died of sepsis and myocarditis and one patient died of pneumonia). The most common categories of immune-related adverse events reported in the relalizumab plus volumab group were hypothyroidism or thyroiditis (in 18.0% of patients), rash (9.3%), and diarrhea or colitis (6.8%) (Tawbi et al., N. Engl. J. Med. , 386: 26-34, 2022).

Given that the optimal targeting mechanisms of immune regulation in the tumor setting are still under exploration, rational new approaches are needed to increase the efficacy of first-line CPI therapy while maintaining reasonable tolerability. D. Study Design and Rationale

Approximately 160 participants were randomized in the study to receive RO7247669 plus nab-paclitaxel (Arm A) or pembrolizumab plus nab-paclitaxel (Arm B) as follows: ● Arm A (n = approximately 80): RO7247669 plus nab-paclitaxel – Participants received blinded RO7247669 every 3 weeks (a “cycle”); plus nab-paclitaxel every week, repeated for 3 weeks, followed by 1 week off, until disease progression or treatment discontinuation. ● Arm B (n = approximately 80 patients): Pembrolizumab plus nab-paclitaxel – Participants received blinded pembrolizumab every 3 weeks (one “cycle”) plus nab-paclitaxel every week for 3 weeks followed by 1 week off until disease progression or treatment discontinuation.

The first approximately 12 participants were enrolled in a safety run-in period during which they were randomized equally to RO7247669 plus nab-paclitaxel or pembrolizumab plus nab-paclitaxel (targeting 6 participants per combination). The remaining participants continued to be randomized in a 1:1 ratio to receive RO7247669 plus nab-paclitaxel or pembrolizumab plus nab-paclitaxel.

For each individual participant, the study is divided into three phases: screening , treatment, and follow-up.

After providing informed consent, patients will be assessed for study eligibility during a 28-day screening period (Days -28 to -1 relative to the first dose of study treatment). Patients who are determined to be eligible based on the screening assessment are randomized to receive study treatment (and are hereafter referred to as “participants”). Patients who do not meet the criteria for participation in the study (screening failures) may be eligible for one rescreening opportunity. Treatment Period

Approximately 160 participants were randomly assigned to each treatment group. Randomization used permuted-block randomization to ensure balanced assignment to each treatment group and was stratified according to the following criteria: prior cancer immunotherapy exposure (yes vs. no) and initial presentation of TNBC as de novo metastatic (stage IV) disease (yes vs. no). No crossover between groups was allowed (i.e., RO7247669 to pembrolizumab or vice versa). Safety Run-In Assessments

An initial safety run-in assessment was conducted to evaluate the safety and tolerability of the new combination of blinded RO7247669 and nab-paclitaxel. The safety run-in assessment was conducted on the Safety Run-In Evaluable Analysis Set (SRAS), defined as at least 12 randomized participants (at least 6 participants in each group) who received at least one dose of the combination treatment. It occurred after SRAS participants had the opportunity to complete 6 weeks of treatment. Screening was held after randomization of the first approximately 12 participants until analysis and clearance for the safety run-in was performed. Assessments

Tumor assessments were performed at baseline and according to the active schedule regardless of treatment dose delays until radiographic disease progression per RECIST v1.1, withdrawal of consent, study discontinuation, or death, whichever occurred first. Participants who underwent palliative therapy for target lesions and were therefore not evaluable for response remained on study and continued to have tumor assessments until disease progression per RECIST v1.1, complete withdrawal of consent, loss to visit, study discontinuation, or death, whichever occurred first.

Participants who discontinued treatment for reasons other than radiographic disease progression per RECIST v1.1 (e.g., toxicity, symptom worsening) continued to have planned tumor assessments according to the active schedule until radiographic disease progression per RECIST v1.1, withdrawal of consent, loss to follow-up, study discontinuation, or death, whichever occurred first, regardless of whether the patient started new anticancer therapy.

Safety assessments at study visits included the incidence, nature, and severity of adverse events, protocol-defined vital signs, laboratory abnormalities, and other protocol-defined tests considered important for the assessment of study safety. The severity of adverse events was assessed according to the National Cancer Institute Common Terminology Criteria for Adverse Events version 5.0 (NCI CTCAE v5.0). The severity of cytokine release syndrome (CRS) was assessed according to the American Society of Transplantation and Cellular Therapy (ASTCT) consensus grading scale.

During the study, samples were collected to monitor serum RO7247669 concentrations and plasma paclitaxel concentrations at specific time points and to detect the presence of antibodies to RO7247669. Samples (including archival and fresh tumor tissue, serum, plasma, and blood samples) were also collected from participants for exploratory biomarker assessments.

All participants who discontinued study treatment were followed for survival until death, loss to follow-up, participant withdrawal, or study termination. Rationale for Patient Selection Using the Investigational Dako PD-L1 IHC 22C3 pharmDx Assay, the Investigational VENTANA PD-L1 (SP142) Assay, and the Investigational VENTANA PD-L1 (SP263) Assay

The Dako PD-L1 IHC 22C3 pharmDx assay, the VENTANA PD-L1 (SP142) assay, and the VENTANA PD-L1 (SP263) assay are intended for use as qualitative immunohistochemistry assays for the assessment of PD-L1 expression in formalin-fixed, paraffin-embedded (FFPE) TNBC tissues. Results from these devices are used to identify PD-L1-positive patients and to support efficacy analyses.

The KEYNOTE-355 study showed that only patients with mTNBC whose tumors expressed PD-L1 with a CPS ≥ 10 (as assessed using the Dako PD-L1 IHC 22C3 pharmDx assay) derived clinical benefit from pembrolizumab (Cortés et al., Lancet, 396: 1817-1828, 2020). Health authorities, including the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA), approved the anti-PD-1 antibody pembrolizumab in combination with chemotherapy for the treatment of patients with advanced TNBC whose tumors are PD-L1 positive, defined as a CPS ≥ 10, as determined by using the Dako PD-L1 IHC 22C3 pharmDx assay.

The IMpassion130 study showed that only patients with mTNBC whose tumors expressed PD-L1 (PD-L1-stained tumor-infiltrating immune cells (ICs) covering ≥ 1% of the tumor area, as assessed by the VENTANA PD-L1 (SP142) assay) derive clinical benefit from the combination of atezolizumab and nab-paclitaxel (Schmid et al., NEJM , 379: 2108-2121, 2018). Health authorities, including the European Medicines Agency (EMA), approved the anti-PD-L1 antibody atezolizumab in combination with nab-paclitaxel for the treatment of patients with advanced TNBC whose tumors are PD-L1 positive, defined as an IC ≥ 1%, as determined by using the VENTANA PD-L1 (SP142) assay.

Diagnostics for RO7247669 were developed using the investigational VENTANA (SP142) assay and the investigational VENTANA (SP263) assay, which has an immunostaining pattern similar to the Dako PD-L1 IHC 22C3 pharmDx assay (Emens et al., J Natl Cancer Inst, 113: 1005-1016, 2021).

The current study randomized individuals with TNBC whose tumor tissue had CPS ≥ 10 or TAP ≥ 5% or IC ≥ 1% as assessed by a central laboratory using the investigational DAKO PD-L1 IHC 22C3 pharmDx, the investigational VENTANA PD-L1 (SP263) assay, and the investigational VENTANA PD-L1 (SP142) assay, respectively. It is expected that there will be some differences in the detection and incidence of these three assays, which will be important to understand from this study. For example, the cutoff for SP263 PD-L1 expression is supported by Roche internal studies of PD-L1 expression, as determined by comparison of the VENTANA SP263 and DAKO 22C3 assays in 694 primary and metastatic TNBC tissue samples, where 50% (344 of 694 patients) had a TAP score ≥ 5%, while the incidence of 22C3 CPS ≥ 10 was 52% (360 of 694 patients). Based on the IMpassion130 study, the incidence of PD-L1 IC ≥ 1% was 41% using the investigational VENTANA SP142 assay (Schmid et al., NEJM , 379: 2108-2121, 2018). Interassay agreement was 77% for samples with a PD-L1 TAP score ≥ 5% by the SP263 assay and PD-L1 CPS ≥ 10 by the 22C3 assay, 73% for samples with a PD-L1 TAP score ≥ 5% by the SP263 assay and PD-L1 IC ≥ 1% by the VENTANA SP142 assay, and 74% for samples with a PD-L1 score ≥ 10% by the 22C3 assay and PD-L1 IC ≥ 1% by the VENTANA SP142 assay (Rugo et al., JNCI , 113(12): 1733-1743, 2021). Rationale for dosing and schedule

The 600 mg RO7247669 Q3W (600 mg on Day 1 of each 21-day cycle) dosing schedule was selected based on existing clinical PK, efficacy, and safety data from the combination Phase Ia/Ib study NP41300. In Study NP41300, the MTD was not reached and no DLTs were observed during dose escalation. Over 90% occupancy of peripheral PD-1 and LAG3 receptors on CD8 ⁺ cells was observed at 50 mg RO7247669 and remained unchanged at all higher doses. In the dose escalation portion of Study NP41300, responses were observed at 600 mg and 2100 mg. Intratumoral PD-1 and LAG3 target engagement was further modeled using the estimated target properties. The 600 mg Q3W dosing regimen was selected to ensure that most participants had at least 90% PD-1 and LAG3 tumor receptor saturation with RO7247669, regardless of intratumor spatial heterogeneity and inter-individual variability. Dosing above 600 mg is not expected to produce additional clinical activity.

In Study NP41300, persistent antidrug antibodies (ADA) developed in approximately 18% of the total population, with an incidence that was apparently not dose-dependent. Exposure effects were observed at doses of 50, 150, and 300 mg; therefore, a 600 mg dose was used in this study to minimize the risk of exposure loss due to ADA.

A participant was considered to have completed the study if he or she completed all periods of the study.

Treatment continued until investigator-assessed disease progression per RECIST v1.1. The total duration of study participation for each individual was expected to range from 1 day to more than 30 months.

The end of the study is defined as the date of the last visit for the last participant in the study or the date the last participant receives the last data point required for statistical analysis or safety follow-up (whichever occurs later). The expected end of the study occurs approximately 30 months after the last participant is randomized. Example 7 : Study Population

In the CO44194 study, approximately 160 patients with metastatic or locally advanced unresectable PD-L1-positive TNBC who had not received prior systemic therapy for the same condition were randomized. Locally advanced disease was not amenable to resection with curative intent, as assessed by the treating investigator/physician. Participants may have received prior chemotherapy and/or cancer immunotherapy in the neoadjuvant or adjuvant setting if all such therapy was completed ≥ 12 months prior to the start of study treatment. Participants must have met all eligibility criteria to be randomized. B. Inclusion Criteria

Potential participants were eligible for inclusion in the study only if all of the following criteria applied: ● Age ≥ 18 years. ● Metastatic or locally advanced unresectable, histologically documented triple-negative breast cancer (TNBC) (absence of HER2 overexpression, ER, and PgR expression by local assessment) ○ HER2 negativity by local laboratory assessment (laboratories are advised to follow the American Society of Clinical Oncology (ASCO)-American College of Pathologists (CAP) HER2 testing guidelines and are explained below): non-expanded in situ hybridization (HER2 to CEP17 ratio ≤ 2.0 or single probe average HER2 gene copy number < 4 signals/cell), or IHC 0. ○ HER2-low status as assessed by local laboratory (laboratories are recommended to follow the ASCO-CAP HER2 testing guidelines, explained below): IHC 2+ and non-expanded in situ hybridization (HER2 to CEP17 ratio < 2.0 or average HER2 gene copy number per probe < 4 signals/cell), or IHC 1+ (and (not required, but if performed) non-expanded in situ hybridization (HER2 to CEP17 ratio < 2.0 or average HER2 gene copy number per probe < 4 signals/cell)). ○ ER and PgR negativity defined as < 1% of cells expressing hormone receptors by IHC analysis, where testing is performed locally. ● Measurable disease per RECIST v1.1. ● Measurable disease outside of bone if metastatic disease (Stage IV). ● Previously irradiated lesions are considered measurable disease only if the same lesion has documented disease progression since radiation. ● No prior systemic therapy for metastatic or locally advanced unresectable TNBC. ○ Radiation therapy for metastatic disease is allowed. There is no minimum run-out period requirement for radiation therapy. Participants should have recovered from the effects of radiation. ○ Prior systemic therapy for early breast cancer (resectable and non-metastatic, in neoadjuvant and/or adjuvant therapy) is allowed. If prior systemic therapy for early TNBC was given, it must have included an anthracycline and a taxane. ○ Prior systemic therapy for early breast cancer is permitted, provided that treatment was completed ≥ 12 months prior to the start of study treatment (Cycle 1, Day 1). ○ Prior anti-PD-1 or anti-PD-L1 therapeutic antibody exposure (e.g., atezolizumab or pembrolizumab) in neoadjuvant and/or adjuvant therapy is permitted, provided that treatment was completed ≥ 12 months prior to the start of study treatment (Cycle 1, Day 1). ● Tumor PD-L1 expression, as documented by central testing of representative tumor tissue specimens. Tumor tissue should be of good quality and assessed for PD-L1 expression in a central laboratory based on total and viable tumor content and must be adjudicated positive as determined using: ○ Investigational VENTANA PD-L1 (SP142) assay (Positive: at least 1% of tumor area occupied by tumor-infiltrating immune cells expressing PD-L1 of any intensity (IC ≥ 1%)) ○ Investigational VENTANA PD-L1 (SP263) assay (Positive: at least 5% of tumor area occupied by tumor or tumor-infiltrating immune cells expressing PD-L1 of any intensity (TAP ≥ 5%)) ○ Investigational Dako PD-L1 IHC 22C3 pharmDx Assays (Positivity: PD-L1 combined positivity score of at least 10 (CPS ≥ 10)) ○ Positivity by any of these assays is sufficient and required for randomization. ● Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. ● Adequate hematologic and end-organ function as defined by the following laboratory test results within 14 days prior to the start of study treatment: ○ Absolute neutrophil count (ANC) ≥ 1.5 × 10 ⁹ /L (1500/µL) without granulocyte colony-stimulating factor (G-CSF) support within the previous 14 days, with one exception: patients with benign ethnic neutropenia (BEN): ANC < 1.3 x 10 ⁹ /L (1300/µL). ○ Lymphocyte count ≥ 0.5 x 10 ⁹ /L (500/µL) ○ Platelet count ≥ 100 × 10 ⁹ /L (100,000/µL) within the past 14 days without transfusion ○ Hemoglobin ≥ 90 g/L (9 g/dL) within the past 14 days without transfusion ○ AST, ALT, and ALP ≤ 2.5 x upper limit of normal (ULN), except for the following: Documented liver metastases: AST and ALT ≤ 5 x ULN. Documented liver or bone metastases: ALP ≤ 5 x ULN. ○ Total bilirubin ≤ 1.5 × ULN, except for the following: Known Gilbert’s disease: Total bilirubin ≤ 3 × ULN. Albumin ≥ 25 g/L (2.5 g/dL). ○ Not on therapeutic anticoagulation: INR and aPTT ≤ 1.5 × ULN. ○ On therapeutic anticoagulation: Stable anticoagulation regimen. ● HIV negative at screening , except for the following: Individuals with a positive HIV test at screening are eligible provided they are stable on antiretroviral therapy, have a CD4 count ≥ 200/µL, and have an undetectable viral load. ● Hepatitis B surface antigen (HBsAg) negative at screening. ● Positive Hepatitis B surface antibody (HBsAb) test at screening, or negative HBsAb at screening and any of the following: ○ Negative Hepatitis B core antibody (HBcAb). ○ Positive HBcAb test followed by quantitative hepatitis B virus (HBV) DNA < 500 IU/mL. ● Negative Hepatitis C virus (HCV) antibody test at screening; or positive HCV antibody test at screening followed by negative HCV RNA test. ● Adequate cardiovascular function: ○ New York Heart Association (NYHA) heart failure ≤ Class 2. ○ Baseline-corrected QT (QTcF) interval ≤ 480 ms. If the QTcF interval is longer than 480 ms but shorter than 500 ms, the participant may undergo a cardiac assessment and, in the absence of clinically significant findings, be considered for treatment. ○ Resting systolic BP ≤ 150 mmHg and diastolic BP 100 mmHg (≤ average of ≥ 3 readings over 2 treatment sessions with brief breaks between each session) (or no clinically significant hypertension). ○ Resting heart rate (HR) between 45 and 100 bpm (or no clinically significant tachycardia). ○ Left ventricular ejection fraction (LVEF) ≥ 50% as assessed by transthoracic echocardiography (TTE) or MUGA (TTE preferred test) within 6 months prior to first dose of study drug. ○ TnT or I (TnI) ≤ institutional upper limit of normal (ULN). Participants with TnT or TnI levels between > 1 and < 2 x ULN will be included in the study if the repeat level is ≤ 1 x ULN. If the repeat level is between ≥ 1 and < 2 x ULN, participants require a cardiac evaluation and may be considered for treatment in the absence of clinically significant findings. ● For female participants of childbearing potential: agree to abstain from sex (avoid heterosexual intercourse) or use contraception and agree not to donate eggs. ● For male participants: agree to abstain from sex (avoid heterosexual intercourse) or use condoms and agree not to donate sperm, as defined below. C. Exclusion Criteria

Potential participants were excluded from the study if any of the following criteria applied: ● Pregnant or breastfeeding, or anticipated pregnancy during the study or within 4 months after the final dose of RO7247669 or pembrolizumab, and within 6 months after the final dose of paclitaxel, pemetrexed, or carboplatin. ● Poor venous circulation ● Glomerular filtration rate (GFR) < 30 mL/min/1.73 ^m2 , as calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. GFR should be assessed by calculation using the CKD-EPI equation: CKD-EPI = 142 x (serum creatinine/A) ^B × 0.9938 ^age x (1.012 if female), where A and B are as follows: ▪ Female serum creatinine ≤ 0.7 mg/dL: A = 0.7 and B = -0.241 ▪ Female serum creatinine > 0.7 mg/dL: A = 0.7 and B = -1.2 ▪ Male serum creatinine ≤ 0.9 mg/dL: A = 0.9 and B = -0.302 ▪ Male serum creatinine > 0.9 mg/dL: A = 0.9 and B= -1.2

To achieve appropriate dosing of nab-paclitaxel, individualized body surface area (BSA)-adjusted GFR values should be used (normalized GFR multiplied by the individual's BSA (calculated using an appropriate formula) and then divided by 1.73). ● History of malignant tumor within 5 years prior to consent, except for cancers being investigated in this study and malignant tumors with negligible risk of metastasis or death (e.g., 5-year OS rate > 90%), such as adequately treated cervical carcinoma in situ, non-melanoma skin cancer, localized prostate cancer, ductal carcinoma in situ, or stage I uterine cancer. ● Symptomatic, untreated, or actively progressive central nervous system (CNS) metastases. Asymptomatic participants with treated CNS lesions were eligible if all of the following criteria were met: ● Measurable disease must be present outside the CNS per RECIST v1.1. ● Participants had no history of intracranial or spinal cord hemorrhage. ● Participants had not received stereotactic radiotherapy within 7 days prior to the start of study treatment, whole brain radiotherapy within 14 days prior to the start of study treatment, or neurosurgery within 28 days prior to the start of study treatment. ● Participants had no ongoing need for corticosteroids as treatment for CNS disease. ● If participants were receiving anticonvulsant therapy, the dose was considered stable (at the discretion of the investigator). ● Metastases limited to the cerebellum or supratentorial regions (i.e., no metastases to the midbrain, pons, medulla, or spinal cord). ● No evidence of significant vasculogenic edema. ● No evidence of transitional progression between completion of CNS-directed therapy and initiation of study treatment (as determined by the investigator). ● Asymptomatic participants with newly discovered CNS metastases at screening are eligible for the study after receiving radiation therapy and/or surgery without repeat screening brain scans. ● History of leptomeningeal disease. ● Pleural effusions, pericardial effusions, or ascites requiring repeated drainage (monthly or more frequently). Participants with indwelling catheters (e.g., PLEURX®) are permitted. ● Hypercalcemia (ionized calcium > 1.5 mmol/L, calcium > 12 mg/dL, or corrected calcium > ULN) or symptomatic hypercalcemia. ● Active or history of autoimmune disease or immunodeficiency, including but not limited to myasthenia gravis, myositis, autoimmune hepatitis, systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, antiphospholipid antibody syndrome, Vegner's granulomatosis (granulomatosis with polyangiitis), Sjögren's syndrome, Guillain-Barré syndrome, or multiple sclerosis, with the following exceptions: ○ Participants with a history of autoimmune-related hypothyroidism who were taking thyroid replacement hormone were eligible for the study. ○ Participants with controlled type 1 diabetes during the insulin regimen were eligible for the study. ○ Participants with eczema, psoriasis, lichen planus simplex, or vitiligo with only dermatologic manifestations (e.g., participants with psoriasis arthritis were excluded) were eligible for the study provided the following criteria were met: – The rash must cover < 10% of the body surface area. – The disease was well controlled at baseline with only low-potency topical corticosteroids. – No acute exacerbation of underlying disease requiring psoralen plus ultraviolet A radiation, methotrexate, retinol, biologics, oral calcineurin inhibitors, high-potency or oral corticosteroids in the 12 months prior to consent. ● History of idiopathic pulmonary fibrosis, organizing pneumonia (e.g., obstructive bronchitis), drug-induced pneumonitis, or idiopathic pneumonia, or evidence of active pneumonia on chest computed tomography (CT) scan. History of radiation pneumonitis (fibrosis) with radiation exposure is permitted. ● Active tuberculosis (as defined by local standards of care). ● Severe cardiovascular/cerebrovascular disease within 3 months prior to consent, including any of the following: hypertensive crisis or encephalopathy, unstable angina, transient ischemic attack or stroke, congestive heart failure, severe arrhythmia requiring treatment (exceptions are atrial tremor, paroxysmal supraventricular tachycardia), history of thromboembolic events (such as myocardial infarction, stroke, or pulmonary embolism). ● History or presence of an abnormal ECG considered clinically significant (e.g., complete left bundle branch block, second or third degree atrioventricular heart block), or evidence of a previous myocardial infarction. ● QT interval corrected using the Fridericia formula (QTcF) > 480 ms, as demonstrated by at least two ECGs taken > 30 minutes apart. If the QTcF interval is longer than 480 ms but shorter than 500 ms, the participant may undergo a cardiac evaluation and, in the absence of clinically significant findings, be considered for treatment. ● History of ventricular arrhythmias or risk factors for ventricular arrhythmias, such as structural heart disease (e.g., severe left ventricular systolic dysfunction, left ventricular hypertrophy), coronary artery disease (with symptoms or diagnostic testing confirming ischemia), clinically significant electrolyte abnormalities (e.g., hypokalemia, hypomagnesemia, hypocalcemia), or family history of unexplained sudden death or long QT syndrome. ● Major surgery within 4 weeks prior to the start of study treatment. Placement of a central venous access catheter (e.g., port or similar catheter) is not considered major surgery. ● Treatment with therapeutic oral or IV antimicrobials (antibacterial, antifungal, antiviral, antiparasitic) within 1 week prior to starting study treatment. Participants receiving prophylactic antibiotics (e.g., to prevent urinary tract infection or exacerbation of chronic obstructive pulmonary disease (COPD)) were eligible for the study. ● Previous allogeneic stem cell or solid organ transplant. ● Use of study medications, any other disease, metabolic disorder, physical examination finding, or clinical laboratory finding that may affect the interpretation of results or may place the participant at high risk for treatment complications are prohibited. ● Treatment with live attenuated vaccines within 28 days prior to starting study medication. ● Study treatment within 28 days prior to the start of study treatment. ● Previous treatment with CD137 agonists or anti-CTLA therapeutic antibodies or anti-LAG3 agents. ● Treatment with systemic immunostimulants (including, but not limited to, interferons and IL-2) within 4 weeks or within 5 drug elimination half-lives (whichever is longer) prior to the start of study treatment. ● Treatment with systemic corticosteroids or other systemic immunosuppressive drugs (including, but not limited to, prednisone, dexamethasone, cyclophosphamide, azathioprine, methotrexate, thalidomide, and anti-TNF agents) within 2 weeks prior to the start of study treatment. ○ Participants who have received acute, low-dose, systemic immunosuppressant medications (e.g., a one-time dose of dexamethasone for nausea, or 48-hour corticosteroids for contrast allergy) are eligible for the study. ○ Participants with a history of allergic reactions to IV contrast media and requiring steroid pretreatment should undergo magnetic resonance imaging (MRI) for baseline and follow-up tumor assessments. ○ Inhaled glucocorticoids are permitted for COPD, mineralocorticoids (e.g., fluocortisol) are permitted for participants with orthostatic hypotension, and low-dose supplemental corticosteroids are permitted for adrenocortical insufficiency. ● History of severe allergic allergic reactions to chimeric or humanized antibodies or fusion proteins. ● Known hypersensitivity to Chinese hamster ovary cell products or to any component of RO7247669 or pembrolizumab formulations. ● Known allergy or hypersensitivity to any component of nab-paclitaxel formulations. Example 8 : Study treatments, other treatments related to the study design, and concomitant therapies

Table 14 provides a description of the assigned study treatments for Study CO44194. Table 14. Study Treatment Description RO7247669 Pembrolizumab Nab-paclitaxel use Experiment Comparison Background treatment Dosage level 600 mg every 3 weeks. No dose reduction is permitted. 200 mg every 3 weeks. No dose reduction is permitted. Initial dose 100 mg/m ² Administration IV infusion IV infusion IV infusion IMP = investigational medicinal product; NIMP = noninvestigational medicinal product.

“Study treatment” means the combination of treatments assigned to a participant as part of this study (i.e., RO7247669 or pembrolizumab and nab-paclitaxel). A. Blinded RO7247669 and Blinded Pembrolizumab

Administration of blinded RO7247669 (600 mg Q3W) or blinded pembrolizumab (200 mg Q3W) by IV infusion was performed in a monitored environment with immediate access to trained personnel and adequate equipment and medications to manage potentially serious reactions.

The initial dose is delivered over 60 (± 15) minutes. If the 60-minute infusion is tolerated without infusion-related adverse events (fever or chills), the second infusion may be delivered over 30 (± 10) minutes. If the 30-minute infusion is well tolerated, all subsequent infusions may be delivered over 30 (± 10) minutes.

Participants who experience an infusion-related adverse event may pre-medicate with antihistamines and/or antipyretics for the next infusion, but the infusion time for that infusion may not be reduced.

Administer RO7247669 and pembrolizumab infusions according to the instructions outlined in Table 15. Table 15. Administration of the First and Subsequent Infusions of RO7247669 or Pembrolizumab First infusion Subsequent infusion RO7247669 or pembrolizumab infusion ● For the first infusion of RO7247669 or pembrolizumab, no premedication is allowed. ● Record vital signs (pulse rate, respiratory rate, blood pressure, temperature, and oxygen saturation) for 60 minutes before the start of the infusion. ● RO7247669 and pembrolizumab are infused over 60 (± 15) minutes. ● Record vital signs every 15 (± 5) minutes during the infusion, if clinically indicated. ● If the patient experienced an infusion-related reaction (IRR) during any prior infusion of RO7247669 or pembrolizumab, premedication with antihistamines and/or antipyretics may be administered for subsequent doses. ● Record vital signs for 60 minutes prior to infusion. ● RO7247669 and pembrolizumab should be infused over 30 (± 10) minutes if the prior infusion was well tolerated without an IRR, or over 60 (± 15) minutes if the patient experienced an IRR during the prior infusion. ● Record vital signs during infusion if clinically indicated. Observation period after RO7247669 or pembrolizumab infusion ● After infusion of RO7247669 or pembrolizumab, patients began a 60-minute observation period. ● Vital signs were recorded at 30 (± 10) minutes after infusion of RO7247669 or pembrolizumab. ● If the patient tolerated the previous RO7247669 or pembrolizumab infusion well without infusion-related adverse events, the observation period after the next and subsequent infusions can be shortened to 30 minutes. If the patient experienced an infusion-related adverse event during the previous infusion, the observation period should be 60 minutes. ● If clinically indicated, record vital signs at 30 (± 10) minutes after the infusion of RO7247669 or pembrolizumab. B. Nab- paclitaxel

The starting dose level of nab-paclitaxel in the study was 100 mg/m ² , given intravenously over 30 minutes weekly, on a repeat schedule of 3 weeks, followed by 1 week off. Doses of nab-paclitaxel were not to be administered more frequently than once every 7 days.

On days when infusion of blinded RO7247669 or pembrolizumab and nab-paclitaxel was planned, chemotherapy was administered after infusion of RO7247669 or pembrolizumab.

Nab-paclitaxel dosing may be adjusted based on local prescribing information or the EUSmPC (in the absence of local approval). Centers follow label guidance to determine nab-paclitaxel dosing for participants experiencing obesity and make dose adjustments as participants change in weight. Infusion sites are closely monitored during drug administration for possible penetration. C. Combination Therapies Permitted Therapies

As described below, the following concomitant therapies are contraindicated: ● Palliative (e.g., treatment of known bone metastases or relief of painful symptoms) local therapies (e.g., surgery, stereotactic radiosurgery, radiation therapy, radiofrequency ablation), as outlined below: Palliative local therapies are permitted provided there is no clear progression per RECIST v1.1. RO7247669 and pembrolizumab may be continued during palliative local therapy. ● Prophylactic or therapeutic anticoagulation (e.g., stable-dose warfarin or low molecular weight heparin) ● Vaccinations (e.g., influenza, COVID-19). Live attenuated vaccines are not permitted. ● Megestrol acetate given as an appetite stimulant. ● Mineralocorticoids (e.g., fluocortisol). ● Inhaled or low-dose corticosteroids for chronic obstructive pulmonary disease or asthma. ● Low-dose mineralocorticoids for orthostatic hypotension or low-dose mineralocorticoids and corticosteroids for adrenocortical insufficiency.

Premedication with nab-paclitaxel was permitted. Premedication with antihistamines, antipyretics, and/or analgesics was permitted only for the second and subsequent RO7247669 and pembrolizumab infusions.

In general, care of participants was managed with supportive therapies, except for those defined as cautious or contraindicated as clinically indicated, in accordance with local standard practice. Participants experiencing infusion-related symptoms could receive symptomatic treatment with acetaminophen, ibuprofen, diphenhydramine, and/or H2 receptor antagonists (e.g., phentermine, ximetidine), or equivalent medications in accordance with local standard practice. Severe infusion-related events manifested by dyspnea, hypotension, wheezing, bronchospasm, tachycardia, decreased oxygen saturation, or respiratory distress should be treated with supportive therapies (e.g., supplemental oxygen and beta-2 adrenergic agonists) as clinically indicated. Use therapy with caution

Systemic corticosteroids, immunosuppressive drugs, and tumor necrosis factor (TNF) inhibitors may attenuate the potential beneficial immunologic effects of treatment with RO7247669 and/or pembrolizumab. Therefore, alternative agents, including antihistamines, should be considered in the setting of conventional administration of systemic corticosteroids, immunosuppressive drugs, or TNF inhibitors. If alternatives are not feasible, systemic corticosteroids, immunosuppressive drugs, and TNF inhibitors may be administered at the investigator's discretion.

Systemic corticosteroids or immunosuppressive drugs are recommended, at the discretion of the investigator, for specific adverse events associated with treatment with RO7247669 and pembrolizumab.

Concomitant use of nab-paclitaxel with drugs known to induce CYP2C8 or CYP3A4 (e.g., rifampin, carbamapine, phenytoin, efavirenz, nevirapine) is not recommended because efficacy may be compromised due to lower paclitaxel exposure.

As described below, the following concomitant therapies are prohibited: ● Investigational therapies within 28 days prior to and during study treatment. ● Concomitant therapies intended to treat cancer (including but not limited to chemotherapy, hormonal therapy, immunotherapy, radiation therapy, and herbal therapies), whether approved by health authorities or experimental (except in certain cases, topical therapies), at various times prior to and during study treatment (depending on the agent) and until disease progression is documented and the participant discontinues study treatment. ● Live attenuated vaccines within 28 weeks prior to initiation of study treatment, during study treatment, and within 5 months after the final dose of study treatment. ● Systemic immune stimulants (including but not limited to interferons and interleukins) within 4 weeks prior to initiation of study treatment or 5 drug elimination half-lives (whichever is longer) and during study treatment, because these agents may increase the risk of autoimmune conditions when given in combination with study treatment. Example 9 : Study Assessments and Procedures A. Efficacy Assessments Tumor and Response Assessments

Participants in the CO44194 study had tumor assessments at screening, every 6 weeks for the first 12 months after treatment initiation, and every 12 weeks thereafter, regardless of dose delay or treatment schedule, until radiographic disease progression according to RECIST v1.1. Therefore, for participants who discontinued treatment for reasons other than radiographic disease progression according to RECIST v1.1, withdrawal of consent, death, or study discontinuation (whichever occurred first), tumor assessments continued as scheduled. In the absence of radiographic disease progression according to RECIST v1.1, tumor assessments continued regardless of whether participants started follow-up anticancer therapy.

All measurable and/or assessable lesions are assessed and recorded during screening. Radiographic Evaluation

Screening evaluations must include a CT scan of the chest, abdomen, and pelvis with contrast (per institutional standard operating procedure). If CT scans with contrast are contraindicated (e.g., in participants with contrast allergy or impaired renal clearance), a CT scan of the chest without contrast and an MRI scan of the abdomen and pelvis (with contrast, if available) may be performed. Response Assessment

Objective responses within individual participants were adjudicated by the investigator at designated time points according to RECIST v1.1. Whenever possible, assessments were performed by the same individual to ensure internal consistency across visits.

Other endpoints (e.g., ORR, PFS, DOR) were calculated programmatically based on investigator response assessments at each specified time point. Example 10 : Study Assessments and Procedures A. Statistical Considerations Statistical Assumptions

The null hypothesis ( _H0 ) and alternative hypothesis ( _H1 ) about PFS in the full analysis set (FAS) of the CO44194 study were tested at a one-sided significance level of 0.1 and can be expressed as the population hazard ratio (HR) of PFS between the experimental group and the control group: H0: ≥ 1 and H1: < 1 Sample size determination

The primary endpoint of progression-free survival (PFS) was analyzed in the FAS. Approximately 160 participants were randomized in a 1:1 ratio to receive RO7247669 plus nab-paclitaxel or pembrolizumab plus nab-paclitaxel.

The study was powered at approximately 74% with a target PFS HR of 0.65 for the primary analysis at an overall alpha of 0.1 (one-sided). Approximately 80 PFS events in the FAS were required for the primary analysis, assuming the following: ● Median PFS was 9.7 months in the pembrolizumab plus nab-paclitaxel arm. ● Target PFS HR of 0.65. ● One-sided log-rank test with a significance level of 0.1. ● 74% power. ● 10% withdrawal rate every 12 months. ● Exponential distribution of survival curves for PFS. ● Accumulation of approximately 160 patients over 15 months.

For 80 PFS events and a target HR of 0.65, an observed HR of 0.751 or less was predicted to result in a statistically significant difference between treatment arms (i.e., an HR of 0.751 was the minimal detectable difference (MDD) for the primary analysis). This corresponded to an increase in median PFS of 3.2 months, from 9.7 months with pembrolizumab plus nab-paclitaxel to 12.9 months with RO7247669 plus nab-paclitaxel.

The primary analysis was performed when approximately 80 PFS events were observed in the FAS and no earlier than 4 months after the last patient was enrolled. The MDD was recalculated based on the actual number of PFS events observed at the time of the primary analysis.

Table 16 shows the efficacy of RO7247669 plus nab-paclitaxel for several possible true potential improvements in PFS. Table 16. Efficacy of Several Possible True Potential Hazard Ratio Values The true potential hazard ratio 0.6 0.65 0.7 Power of the log-rank ^testa 84% 74% 63% The ^a -power was calculated using a one-sided α = 0.1 for the analysis set

The participant analysis set used for analysis purposes is defined in Table 17. Table 17. Participant Analysis Set Participant Analysis Set describe FAS All randomized participants, regardless of whether they received the assigned treatment. SAS Participants who were randomized and received at least one dose of any study treatment. 22C3-Positive Analysis Set All randomized participants with positive PD-L1 expression as determined by the investigational Dako PD-L1 IHC 22C3 pharmDx assay (CPS ≥ 10) SP142-Positive Analysis Set All randomized participants with positive PD-L1 expression as determined by the investigational VENTANA PD-L1 (SP142) assay (IC ≥ 1%) SP263-Positive Analysis Set All randomized participants with positive PD-L1 expression as determined by the investigational VENTANA PD-L1 (SP263) assay (TAP ≥ 5%) PK Evaluable Set Participants who received at least one dose of study treatment and had at least one post-dose evaluable PK sample Safety Import Evaluable Analysis Set Minimum 12 participants randomized to receive at least one dose of the combination therapy CPS = combined positivity score; FAS = full analysis set; IC = tumor-infiltrating immune cells; SAS = safety analysis set; PD-L1 = programmed death-ligand 1; PK = pharmacokinetic; TAP = tumor area positivity. B. General Considerations for Statistical Analysis

Analyses of PFS, ORR, and OS were performed in the FAS, 22C3-positive analysis set, SP142-positive analysis set, and SP263-positive analysis set. Other efficacy endpoints were assessed in the FAS unless otherwise specified. Participants were assigned according to the treatment assigned at randomization, regardless of whether they received any assigned study drug.

Safety analyses were performed on the SAS unless otherwise specified. Participants were assigned to the groups according to the actual treatment they received. Specifically, if a participant received any amount of RO7247669, that participant was assigned to the RO7247669 plus nab-paclitaxel group for the safety analyses, regardless of the initial treatment assignment at randomization. Methods for Estimation of the Primary End Points

The primary efficacy endpoint was investigator-assessed PFS in the FAS. Hypothesis testing for PFS was performed at a one-sided significance level of 0.1.

PFS was defined as the time from randomization to the first occurrence of disease progression (as assessed by the investigator using RECIST v1.1) or death from any cause, whichever occurred first. Data for participants who did not experience disease progression or death as of the clinical cutoff date were censored at the date of last tumor assessment. Participants who did not have a post-baseline tumor assessment were censored at the date of randomization.

PFS was compared between treatment groups using a stratified log-rank test. HRs were estimated using a stratified Cox proportional hazards model. 95% CIs for HRs are provided. Stratification factors were the same as those used for randomization (ie, prior cancer immunotherapy exposure and initial presentation of TNBC as de novo metastatic (stage IV) disease). Stratification factors were removed from the stratified analysis if there was a risk of over-stratification. Results from the unstratified analysis are also presented.

The median PFS (if achieved) for each treatment group will be estimated using the Kaplan-Meier method and Kaplan-Meier curves will be generated. The 95% CI for the median PFS (if achieved) for each treatment group will be constructed using the Brookmeyer-Crowley method (Brookmeyer and Crowle 1982). Methods for Estimation of Secondary Endpoints

PFS was analyzed in the 22C3-positive analysis set, SP142-positive analysis set, and SP263-positive analysis set at the same time as FAS using the same methods as above.

Confirmed ORR was assessed in the FAS, 22C3-positive analysis set, SP142-positive analysis set, and SP263-positive analysis set.

Confirmed objective response rate was defined as two consecutive complete responses (CR) or partial responses (PR) assessed by the investigator ≥ 4 weeks after randomization according to RECIST v1.1. Participants who did not meet these criteria, including those who did not have any post-baseline tumor assessments, were considered non-responders.

Confirmed ORR was defined as the proportion of participants with a confirmed objective response after randomization. ORR analyses were performed in participants with measurable disease at baseline and compared between treatment groups using the stratified Cochran Mantel-Haenszel test. Stratification factors were the same as those described for the PFS primary endpoint analysis. Results for unstratified analyses are also presented. The difference in ORR between treatment groups will be calculated, and its 95% CI will be calculated using the Newcombe method. ORR estimates for each treatment group will be calculated, and their 95% CI will be calculated using the Wilson scoring method.

Duration of response (DOR) was assessed on the FAS. DOR was defined as the interval from the date of the first confirmed objective response to the date of the first occurrence of disease progression as determined by the investigator according to RECIST v1.1 or death from any cause, whichever occurred first. Confirmed DOR was assessed in the subset of participants with measurable disease at baseline who achieved a confirmed objective response as assessed by the investigator according to RECIST v1.1. Data for participants who did not experience disease progression or death were censored at the date of the last tumor assessment.

OS was assessed in the FAS, 22C3-positive analysis set, SP142-positive analysis set, and SP263-positive analysis set.

OS is the time from randomization to death from any cause. Data for participants whose deaths were not reported at the time of analysis were censored at the date last known to be alive. Data for participants without postbaseline information were censored at the date of randomization. Comparisons of OS between treatment groups were performed in the same manner as for treatment comparisons for the primary endpoint of PFS. Median OS and corresponding 95% CIs (if reached) were estimated using the same methods as those used for PFS.

The PFS rate at 12 months for each treatment group was estimated using the Kaplan-Meier method, and the 95% CI was calculated using the standard errors derived from the Greenwood formula. The 95% CI for the difference in PFS rate between the two treatment groups was estimated using the normal approximation method. A similar analysis was performed for the OS rate 12 months after randomization. Safety Analysis

Safety analyses included all randomized patients who received at least one dose of study treatment, with patients divided according to the actual treatment received.

Safety was assessed by adverse events, changes in laboratory test results, changes in vital signs, exposure to study treatment, and was presented by study group.

Verbatim descriptions of adverse events were mapped to MedDRA index terms. The severity of all adverse events was graded by the investigator according to the NCI CTCAE v5.0, and the severity of CRS was graded by the investigator according to the American Society for Transplantation and Cellular Therapy (ASTCT) consensus grading scale. All adverse events occurring during or after the first dose of study drug, serious adverse events, adverse events leading to death, adverse events of special concern, and adverse events leading to discontinuation of study treatment (i.e., treatment-emergent adverse events) were summarized by mapped term, appropriate thesaurus level, and severity grade. For events of varying severity, the highest grade was used in the summary. Deaths and causes of death were summarized.

Display relevant laboratory test and vital sign data by time and, where appropriate, by grade. In addition, summarize the highest severity grade at baseline and after baseline using a shift table for selected laboratory tests. Summarize changes in vital signs. Pharmacokinetic analysis

Individual and mean serum RO7247669 concentration versus time data were tabulated and plotted by dose level. Serum pharmacokinetics of RO7247669 were summarized by estimating total exposure (area under the concentration-time curve), maximum observed concentration (serum), total clearance, steady-state distribution volume, and terminal drug elimination half-life (as appropriate for the data collected). Estimates of these parameters were tabulated and summarized (mean, standard deviation, coefficient of variation, median, and range). Inter-participant variability and drug accumulation were assessed.

Perform additional PK analyses as appropriate.

Tabulate and report the pharmacokinetic data of paclitaxel. Interim analysis

The interim analysis occurred after approximately 80 randomized patients had a minimum follow - up of 3 months . SEQ ID NO. sequence Sequence Type Type of organism 1 GFSFSSY Protein Synthetic constructs 2 TGRVYFALD Protein Synthetic constructs 3 SESVDTSDNSF Protein Synthetic constructs 4 NYDVPW Protein Synthetic constructs 5 EVQLLESGGGLVQPGGSLRLSCAASGFSFSSYTMSWVRQAPGKGLEWVATISGGGRDIYYPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVLLTGRVYFALDSWGQGTLVTVSS Protein Synthetic constructs 6 DIVMTQSPDSLAVSLGERATINCKASESVDTSDNSFIHWYQQKPGQSPKLLIYRSSTLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQNYDVPWTFGQGTKVEIK Protein Synthetic constructs 7 DYTMN Protein Synthetic constructs 8 VISWDGGGTYYTDSVKG Protein Synthetic constructs 9 GLTDTTLYGSDY Protein Synthetic constructs 10 RASQSISSYLN Protein Synthetic constructs 11 AASTLQS Protein Synthetic constructs 12 QQTYSSPLT Protein Synthetic constructs 13 EVQLLESGGGLVQPGGSLRLSCAASGFIFDDYTMNWVRQAPGKGLEWVAVISWDGGGTYYTDSVKGRFTISRDDFKNTLYLQMNSLRAEDTAVYYCAKGLTDTTLYGSDYWGQGTLVTVSS Protein Synthetic constructs 14 DIQMTQSPSSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSSPLTFGGGTKVEIK Protein Synthetic constructs 15 DIVMTQSPDSLAVSLGERATINCKASESVDTSDNSFIHWYQQKPGQSPKLLIYRSSTLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQNYDVPWTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK Protein Synthetic constructs 16 EVQLLESGGGLVQPGGSLRLSCAASGFSFSSYTMSWVRQAPGKGLEWVATISGGGRDIYYPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVLLTGRVYFALDSWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGEC Protein Synthetic constructs 17 EVQLLESGGGLVQPGGSLRLSCAASGFIFDDYTMNWVRQAPGKGLEWVAVISWDGGGTYYTDSVKGRFTISRDDFKNTLYLQMNSLRAEDTAVYYCAKGLTDTTLYGSDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEK VEPKSCD KTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK Protein Synthetic constructs 18 DIQMTQSPSSSLSASVGDRVTITCRASQSISLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSSPLTFGGGTKVEIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS SPVTKSFNRGEC Protein Synthetic constructs 19 MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTG QPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL Protein Homo sapiens 20 VPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGF NVSIMYNLTVLGLEPPTPLTVYAGAGS RVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLLFLILGVLSLLLLVTGAFGFHLWRRQWRPRRFSALEQGI HPPQAQSKIEELEQEPEPEPEPEPEPEPEPEQL Protein Homo sapiens twenty one VPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGF NVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHL Protein Homo sapiens twenty two GGGGS Protein Synthetic constructs twenty three GGGGSGGGGS Protein Synthetic constructs twenty four SGGGGSGGGG Protein Synthetic constructs 25 GGGGSGGGGSGGGG Protein Synthetic constructs 26 GSPGSSSSGS Protein Synthetic constructs 27 GGGGSGGGGSGGGGS Protein Synthetic constructs 28 GGGGSGGGGSGGGGSGGGGS Protein Synthetic constructs 29 GSGSGSGS Protein Synthetic constructs 30 GSGSGNGS Protein Synthetic constructs 31 GGSGSGSG Protein Synthetic constructs 32 GGSGSG Protein Synthetic constructs 33 GGS Protein Synthetic constructs 34 GGSGNGSG Protein Synthetic constructs 35 GGNGSGSG Protein Synthetic constructs 36 GGNGSG Protein Synthetic constructs 37 DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKGGG GSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFIFDDYTMNWVRQAPGKGLEWVAVISWDGGGTYYTDSVKGRFTISRDDFKNTLYLQMNSLRAEDTAVYYCAKGLTDTTLYGSDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDEKVEPKSC Protein Synthetic constructs 38 MRIFAVFFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNE IFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET Protein Homo sapiens VIII. Other embodiments

Some embodiments of the technology described herein may be defined according to any of the following numbered paragraphs: 1. A method for treating an individual having cancer (e.g., non-squamous non-small cell lung cancer (NSCLC)), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; and one or more chemotherapeutic agents, for example: (b) an anti-metabolic drug (e.g., pemetrexed); and (c) a platinum-based chemotherapeutic agent (e.g., carboplatin). 2. The method of paragraph 1, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 3. The method of paragraphs 1 or 2, wherein the method comprises administering pemetrexed to the subject at a dose of about 500 mg/m ² (e.g., at a dose of 500 mg/m ² ) every three weeks. 4. The method of any one of paragraphs 1 to 3, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of about 5 mg/mL • min (e.g., a target AUC of 5 mg/mL • min) every three weeks. 5. The method of any of paragraphs 1 to 4, wherein the length of each of the one or more dosing cycles is about 21 days (e.g., is 21 days), and wherein the method comprises administering the bispecific antibody, pemetrexed, and carboplatin to the individual on about day 1 (e.g., day 1 ± 1 day) of each of the one or more dosing cycles. 6. The method of any one of paragraphs 1 to 5, wherein the method comprises (a) first administering to the subject a bispecific antibody, secondly administering an anti-metabolic drug (e.g., pemetrexed), and thirdly administering a platinum-based chemotherapy (e.g., carboplatin), or (b) first administering to the subject an anti-metabolic drug (e.g., pemetrexed), secondly administering a platinum-based chemotherapy (e.g., carboplatin), and thirdly administering the bispecific antibody. 7. The method of any one of paragraphs 1 to 6, wherein the method comprises intravenously administering to the subject a bispecific antibody, an anti-metabolic drug (e.g., pemetrexed), and a platinum-based chemotherapy (e.g., carboplatin). 8. The method of paragraph 7, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) the anti-metabolic drug (e.g., pemetrexed) is administered over 10 minutes; and/or (d) the platinum-based chemotherapy (e.g., carboplatin) is administered over 30 to 60 minutes. 9. The method of any one of paragraphs 1 to 8, wherein the dosing regimen comprises four dosing cycles. 10. The method of any of paragraphs 1 to 9, wherein the dosing regimen further comprises one or more additional dosing cycles of (a) the bispecific antibody and (b) an anti-metabolic drug (e.g., pemetrexed). 11. The method of paragraph 10, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of (a) the bispecific antibody and (b) an anti-metabolic drug (e.g., pemetrexed). 12. A method of treating an individual with cancer (e.g., squamous NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3; and one or more chemotherapeutic agents, such as: (b) a taxane (e.g., paclitaxel); and (c) a platinum-based chemotherapeutic agent (e.g., carboplatin). 13. The method of paragraph 12, wherein the method comprises administering to the individual the bispecific antibody at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 14. The method of paragraph 12 or 13, wherein the method comprises administering paclitaxel to the subject at a dose of about 200 mg/m ² (e.g., a dose of 200 mg/m ² ) every three weeks. 15. The method of any one of paragraphs 12 to 14, wherein the method comprises administering carboplatin to the subject at a target AUC of about 5 mg/mL • min (e.g., a target AUC of 5 mg/mL • min) every three weeks. 16. The method of any of paragraphs 12 to 15, wherein the length of each of the one or more dosing cycles is about 21 days (e.g., is 21 days), and wherein the method comprises administering to the individual the bispecific antibody, the taxane (e.g., paclitaxel), and the platinum-based chemotherapy (e.g., carboplatin) on about day 1 (e.g., day 1±1 day) of each of the one or more dosing cycles. 17. The method of any one of paragraphs 12 to 16, wherein the method comprises (a) administering the bispecific antibody first to the subject, administering the taxane (e.g., paclitaxel) second, and administering the platinum-based chemotherapy (e.g., carboplatin) third, or (b) administering the taxane (e.g., paclitaxel) first to the subject, administering the platinum-based chemotherapy (e.g., carboplatin) second, and administering the bispecific antibody third. 18. The method of any one of paragraphs 12 to 17, wherein the method comprises administering the bispecific antibody, the taxane (e.g., paclitaxel) and the platinum-based chemotherapy (e.g., carboplatin) intravenously to the subject. 19. The method of paragraph 18, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) the taxane (e.g., paclitaxel) is administered over 3 hours; and/or (d) the platinum-based chemotherapy (e.g., carboplatin) is administered over 30 to 60 minutes. 20. The method of any one of paragraphs 12 to 19, wherein the dosing regimen comprises four dosing cycles. 21. The method of any of paragraphs 12 to 20, wherein the dosing regimen further comprises one or more additional dosing cycles of the bispecific antibody. 22. The method of paragraph 21, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody. 23. The method of any of paragraphs 12 to 22, wherein the subject is premedicated with oral or IV steroids and antihistamines prior to administration of the taxane (e.g., paclitaxel). 24. The method of any of paragraphs 1 to 23, wherein the cancer is stage IIIB/IIIC or stage IV (e.g., the NSCLC is: (a) stage IIIB/IIIC NSCLC; or (b) stage IV NSCLC). 25. The method of any of paragraphs 1 to 24, wherein the individual has not received prior systemic therapy for metastatic cancer (e.g., metastatic NSCLC). 26. The method of any of paragraphs 1 to 25, wherein the individual has not been previously treated with immune checkpoint blockade therapy. 27. The method of any of paragraphs 1 to 26, wherein the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with an Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent. 28. The method of any of paragraphs 1 to 27, wherein the individual has not been previously treated with a CD137 agonist. 29. The method of any of paragraphs 1 to 28, wherein the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases. 30. The method of any of paragraphs 1 to 29, wherein the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or an anaplastic lymphoma kinase ( ALK ) fusion oncogene. 31. The method of any of paragraphs 1 to 30, wherein the individual has an Eastern Cooperative Oncology (ECOG) performance status of 0 or 1. 32. The method of any one of paragraphs 1 to 31, wherein the PD-L1 tumor proportion score (TPS) or the amount of PD-L1 expression on tumor cells (TC) of a tumor sample from a cancer (e.g., NSCLC) of the individual is < 1%. 33. The method of any one of paragraphs 1 to 31, wherein the PD-L1 TPS or the amount of PD-L1 expression on TC of a tumor sample from a cancer (e.g., NSCLC) of the individual is between 1% and 49%. 34. The method of any one of paragraphs 1 to 31, wherein the PD-L1 TPS or the amount of PD-L1 expression on TC of a tumor sample from a cancer (e.g., NSCLC) of the individual is ≥ 50%. 35. The method of any of paragraphs 1 to 34, wherein the method increases progression-free survival (PFS) as compared to a reference PFS. 36. The method of paragraph 35, wherein the reference PFS is the PFS of a population of individuals who have received a control therapy. 37. The method of any of paragraphs 1 to 36, wherein the method increases the objective response rate (ORR) of a population of individuals treated according to the method as compared to a reference ORR. 38. The method of paragraph 37, wherein the reference ORR is the ORR of a population of individuals who have received a control therapy. 39. The method of any of paragraphs 1 to 38, wherein the method increases the OS as compared to a reference overall survival (OS). 40. The method of paragraph 39, wherein the reference OS is the OS of a population of individuals who have received a control therapy. 41. The method of any of paragraphs 1 to 40, wherein the method results in an increase in duration of response (DOR) compared to a reference DOR. 42. The method of paragraph 41, wherein the reference DOR is the DOR of a population of subjects who have received a control therapy. 43. The method of any of paragraphs 36 to 42, wherein: (a) the control therapy comprises pembrolizumab, an anti-metabolic drug (e.g., pemetrexed) and a platinum-based chemotherapy (e.g., carboplatin) and does not comprise a bispecific antibody (e.g., the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, an anti-metabolic drug (e.g., pemetrexed) and a platinum-based chemotherapy (e.g., carboplatin) and does not comprise a bispecific antibody); or (b) the control therapy comprises pembrolizumab, a taxane (e.g., paclitaxel) and a platinum-based chemotherapy (e.g., carboplatin) and does not include a bispecific antibody (e.g., the individual has non-squamous NSCLC and the control therapy includes pembrolizumab, a taxane (e.g., paclitaxel) and a platinum-based chemotherapy (e.g., carboplatin) and does not include a bispecific antibody). 44. The method of any one of paragraphs 1 to 43, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) a highly variable region H1 (HVR-H1) sequence comprising the amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising the amino acid sequence GGR; and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3; (ii) an HVR-L2 sequence comprising the amino acid sequence RSS; and (iii) an HVR-L3 sequence comprising SEQ ID NO: 45. The method of paragraph 44, wherein the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. 46. The method of paragraph 45, wherein the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. 47. The method of any of paragraphs 45 or 46, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. 48. The method of paragraph 47, wherein the Fc receptor is an Fcγ receptor. 49. The method of any one of paragraphs 44 to 48, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10; (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11; and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12. 50. The method of any one of paragraphs 44 to 49, wherein the first antigen-binding fragment comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14. 51. The method of any one of paragraphs 44 to 50, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the association of the first unit of the Fc domain with the second unit. 52. The method of any one of paragraphs 45 to 51, wherein the first unit of the Fc domain comprises amino acid substitutions S354C and T366W (numbered according to the Kabat EU index), and the second unit of the Fc domain comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to the Kabat EU index). 53. The method of any one of paragraphs 44 to 52, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and a second Fab fragment comprising the second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14, and an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to Kabat EU index numbering). 54. The method of paragraph 53, wherein in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. 55. The method of paragraph 54, wherein in the first Fab fragment, the variable domains VL and VH are replaced with each other. 56. The method of any one of paragraphs 53 to 55, wherein in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 57. The method of paragraph 56, wherein in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 58. The method of any one of paragraphs 44 to 57, wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. 59. The method of paragraph 58, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18. 60. A method for treating an individual having cancer (e.g., non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (1) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising SEQ ID NO: 17 ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) an anti-metabolic drug (e.g., pemetrexed), wherein in the aspect of the anti-metabolic drug being pemetrexed, the method comprises administering pemetrexed to the individual at a dose of 500 mg/m ² every three weeks; and (c) a platinum-based chemotherapeutic agent (e.g., carboplatin), wherein in the aspect of the platinum-based chemotherapeutic agent being carboplatin, the method comprises administering carboplatin to the individual at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks; or (2) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a PD-1- and a LAG3-targeting antibody. and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering the bispecific antibody to an individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, comprising the following hypervariable regions (HVRs): an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising SEQ ID NO: 4 an amino acid sequence of SEQ ID NO: 7; and (ii) a second antigen-binding domain that specifically binds to LAG3, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; (b) an anti-metabolic drug (e.g., pemetrexed), wherein in the aspect that the anti-metabolic drug is pemetrexed, the method comprises administering 500 mg/m every three weeks ² ; and (c) a platinum-based chemotherapy agent (e.g., carboplatin), wherein in the aspect where the platinum-based chemotherapy agent is carboplatin, the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of 5 mg/mL•min every three weeks. 61. A method for treating an individual having cancer (e.g., squamous stage IIIB/IIIC NSCLC or stage IV NSCLC), the method comprising administering to the individual a dosing regimen comprising one or more dosing cycles of: (1) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) a taxane (e.g., paclitaxel), wherein in the aspect of the taxane being paclitaxel, the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) a platinum-based chemotherapeutic agent (e.g., carboplatin), wherein in the aspect of the platinum-based chemotherapeutic agent being carboplatin, the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL • min every three weeks; or (2) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3 The method comprises administering the bispecific antibody to an individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4; and (ii) a first antigen-binding domain that specifically binds to PD-1, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4 a second antigen-binding domain that specifically binds to leukemia/antigens, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; (b) a taxane (e.g., paclitaxel), wherein in the aspect that the taxane is paclitaxel, the method comprises administering paclitaxel to the individual at a dose of 200 mg/ ^m2 every three weeks; and (c) A platinum-based chemotherapeutic agent (eg, carboplatin), wherein in the aspect where the platinum-based chemotherapeutic agent is carboplatin, the method comprises administering carboplatin to the subject at a target AUC of 5 mg/mL•min every three weeks. 62. The method of any one of paragraphs 1 to 61, wherein the subject is a human. 63. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with cancer (e.g., non-squamous non-small cell lung cancer (NSCLC)), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents, wherein the bispecific antibody comprises a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; and one or more chemotherapeutic agents, such as: (b) an anti-metabolic drug (e.g., pemetrexed); and (c) 64. The bispecific antibody of paragraph 63, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 65. The bispecific antibody of paragraph 63 or 64, wherein the method comprises administering pemetrexed to the subject at a dose of about 500 mg/m ² (e.g., at a dose of 500 mg/m ² ) every three weeks. 66. The bispecific antibody of any of paragraphs 63 to 65, wherein the method comprises administering carboplatin to the subject at a target area under the concentration-time curve (AUC) of about 5 mg/mL • min (e.g., a target AUC of 5 mg/mL • min) every three weeks. 67. The bispecific antibody of any of paragraphs 63 to 66, wherein the length of each of the one or more dosing cycles is about 21 days (e.g., is 21 days), and wherein the method comprises administering the bispecific antibody, pemetrexed, and carboplatin to the subject on about day 1 (e.g., day 1 ± 1 day) of each of the one or more dosing cycles. 68. The bispecific antibody of any one of paragraphs 63 to 67, wherein the method comprises (a) administering the bispecific antibody to the individual first, administering an anti-metabolic drug (e.g., pemetrexed) second, and administering a platinum-based chemotherapy (e.g., carboplatin) third, or (b) administering an anti-metabolic drug (e.g., pemetrexed) to the individual first, administering a platinum-based chemotherapy (e.g., carboplatin) second, and administering the bispecific antibody third. 69. The bispecific antibody of any one of paragraphs 63 to 68, wherein the method comprises administering the bispecific antibody, an anti-metabolic drug (eg, pemetrexed) and a platinum-based chemotherapy agent (eg, carboplatin) intravenously to the individual. 70. The bispecific antibody of paragraph 69, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) the anti-metabolic drug (e.g., pemetrexed) is administered over 10 minutes; and/or (d) the platinum-based chemotherapy agent (e.g., carboplatin) is administered over 30 to 60 minutes. 71. The bispecific antibody of any one of paragraphs 63 to 70, wherein the dosing regimen comprises four dosing cycles. 72. The bispecific antibody of any one of paragraphs 63 to 71, wherein the dosing regimen further comprises one or more additional dosing cycles of (a) the bispecific antibody and (b) an anti-metabolic drug (e.g., pemetrexed). 73. The bispecific antibody of paragraph 72, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of (a) the bispecific antibody and (b) an anti-metabolic drug (e.g., pemetrexed). 74. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with cancer (e.g., squamous NSCLC), wherein the method comprises a dosing regimen comprising a bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more dosing cycles of: (a) the bispecific antibody; and one or more chemotherapeutic agents, such as: (b) a taxane (e.g., paclitaxel); and (c) 75. The bispecific antibody of paragraph 74, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 76. The bispecific antibody of paragraph 74 or 75, wherein the method comprises administering paclitaxel to the subject at a dose of about 200 mg/m ² (e.g., at a dose of 200 mg/m ² ) every three weeks. 77. The bispecific antibody of any one of paragraphs 74 to 76, wherein the method comprises administering carboplatin to the subject at a target AUC of about 5 mg/mL • min (e.g., a target AUC of 5 mg/mL • min) every three weeks. 78. The bispecific antibody of any one of paragraphs 74 to 77, wherein the length of each of the one or more dosing cycles is about 21 days (e.g., is 21 days), and wherein the method comprises administering the bispecific antibody, a taxane (e.g., paclitaxel), and a platinum-based chemotherapy (e.g., carboplatin) to the subject on about day 1 (e.g., day 1 ± 1 day) of each of the one or more dosing cycles. 79. The bispecific antibody of any one of paragraphs 74 to 78, wherein the method comprises (a) administering to the subject a taxane (e.g., paclitaxel) secondarily and a platinum-based chemotherapy (e.g., carboplatin) thirdly, or (b) administering to the subject a taxane (e.g., paclitaxel) first, a platinum-based chemotherapy (e.g., carboplatin) secondarily, and the bispecific antibody thirdly. 80. The bispecific antibody of any one of paragraphs 74 to 79, wherein the method comprises administering to the subject intravenously a taxane (e.g., paclitaxel) and a platinum-based chemotherapy (e.g., carboplatin). 81. The bispecific antibody of paragraph 80, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles; (c) the taxane (e.g., paclitaxel) is administered over 3 hours; and/or (d) the platinum-based chemotherapy (e.g., carboplatin) is administered over 30 to 60 minutes. 82. The bispecific antibody of any one of paragraphs 74 to 80, wherein the dosing regimen comprises four dosing cycles. 83. The bispecific antibody of any one of paragraphs 74 to 82, wherein the dosing regimen further comprises one or more additional dosing cycles of the bispecific antibody. 84. The bispecific antibody of paragraph 83, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody. 85. The bispecific antibody of any one of paragraphs 74 to 84, wherein the subject is premedicated with oral or IV steroids and antihistamines prior to administration of a taxane (e.g., paclitaxel). 86. The bispecific antibody of any of paragraphs 63 to 85, wherein the cancer is stage IIIB/IIIC or stage IV (e.g., the NSCLC is: (a) stage IIIB/IIIC NSCLC; or (b) stage IV NSCLC). 87. The bispecific antibody of any of paragraphs 63 to 86, wherein the individual has not received prior systemic therapy for metastatic cancer (e.g., metastatic NSCLC). 88. The bispecific antibody of any of paragraphs 63 to 87, wherein the individual has not been previously treated with immune checkpoint blockade therapy. 89. The bispecific antibody of any of paragraphs 63 to 88, wherein the individual has not been previously treated with an anti-cytotoxic T lymphocyte-associated protein 4 (CTLA4) agent, an anti-T cell immune receptor with an Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent. 90. The bispecific antibody of any of paragraphs 63 to 89, wherein the individual has not been previously treated with a CD137 agonist. 91. The bispecific antibody of any of paragraphs 63 to 90, wherein the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases. 92. The bispecific antibody of any of paragraphs 63 to 91, wherein the individual does not have a known mutation in the epidermal growth factor receptor ( EGFR ) gene or anaplastic lymphoma kinase ( ALK ) fusion oncogene. 93. The bispecific antibody of any of paragraphs 63 to 92, wherein the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. 94. The bispecific antibody of any one of paragraphs 63 to 93, wherein the PD-L1 tumor proportion score (TPS) or the PD-L1 expression on tumor cells (TC) of a tumor sample from a cancer (e.g., NSCLC) of the individual is < 1%. 95. The bispecific antibody of any one of paragraphs 63 to 93, wherein the PD-L1 TPS or the PD-L1 expression on TC of a tumor sample from a cancer (e.g., NSCLC) of the individual is between 1% and 49%. 96. The bispecific antibody of any of paragraphs 63 to 93, wherein the expression of PD-L1 on the PD-L1 TPS or TC of a tumor sample from a cancer (e.g., NSCLC) of the individual is ≥50%. 97. The bispecific antibody of any of paragraphs 63 to 96, wherein the method increases progression-free survival (PFS) compared to a reference PFS. 98. The bispecific antibody of paragraph 97, wherein the reference PFS is the PFS of a population of individuals who have received control therapy. 99. The bispecific antibody of any of paragraphs 63 to 98, wherein the method increases the objective response rate (ORR) of a population of individuals treated according to the method compared to a reference ORR. 100. The bispecific antibody of paragraph 99, wherein the reference ORR is the ORR of a population of individuals who have received control therapy. 101. The bispecific antibody of any of paragraphs 63 to 100, wherein the method results in an increase in OS compared to a reference overall survival (OS). 102. The bispecific antibody of paragraph 101, wherein the reference OS is the OS of a population of individuals who have received control therapy. 103. The bispecific antibody of any of paragraphs 63 to 102, wherein the method results in an increase in DOR compared to a reference duration of response (DOR). 104. The bispecific antibody of paragraph 103, wherein the reference DOR is the DOR of a population of individuals who have received control therapy. 105. The bispecific antibody of any of paragraphs 98 to 104, wherein: (a) the control therapy comprises pembrolizumab, an anti-metabolic drug (e.g., pemetrexed) and a platinum-based chemotherapy (e.g., carboplatin) and does not comprise the bispecific antibody (e.g., the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, an anti-metabolic drug (e.g., pemetrexed) and a platinum-based chemotherapy (e.g., carboplatin) and does not comprise the bispecific antibody); or (b) the control therapy comprises pembrolizumab, a taxane (e.g., paclitaxel) and a platinum-based chemotherapy (e.g., carboplatin) and does not include a bispecific antibody (e.g., the individual has non-squamous NSCLC and the control therapy includes pembrolizumab, a taxane (e.g., paclitaxel) and a platinum-based chemotherapy (e.g., carboplatin) and does not include a bispecific antibody). 106. The bispecific antibody of any one of paragraphs 63 to 105, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) a highly variable region H1 (HVR-H1) sequence comprising the amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising the amino acid sequence GGR; and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3; (ii) an HVR-L2 sequence comprising the amino acid sequence RSS; and (iii) HVR-L3 sequence, which comprises the amino acid sequence of SEQ ID NO: 4. 107. The bispecific antibody of paragraph 106, wherein the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. 108. The bispecific antibody of paragraph 107, wherein the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. 109. The bispecific antibody of any one of paragraphs 107 or 108, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. 110. The bispecific antibody of paragraph 109, wherein the Fc receptor is an Fcγ receptor. 111. The bispecific antibody of any one of paragraphs 106 to 110, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12. 112. The bispecific antibody of any one of paragraphs 106 to 111, wherein the first antigen-binding fragment comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14. 113. The bispecific antibody of any one of paragraphs 106 to 112, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the association of the first unit and the second unit of the Fc domain. 114. The bispecific antibody of any one of paragraphs 107 to 113, wherein the first Fc domain unit comprises amino acid substitutions S354C and T366W (numbered according to the Kabat EU index), and the second Fc domain unit comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to the Kabat EU index). 115. The bispecific antibody of any one of paragraphs 106 to 114, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising the second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14; and an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (According to the Kabat EU index numbering). 116. The bispecific antibody of paragraph 115, wherein in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. 117. The bispecific antibody of paragraph 116, wherein in the first Fab fragment, the variable domains VL and VH are replaced with each other. 118. The bispecific antibody of any one of paragraphs 115 to 117, wherein in the constant domain CL of one of said Fab fragments, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (numbering according to the Kabat EU index), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (numbering according to the Kabat EU index). 119. A bispecific antibody as described in paragraph 118, wherein in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (numbering according to the Kabat EU index), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (numbering according to the Kabat EU index). 120. The bispecific antibody of any one of paragraphs 106 to 119, wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. 121. The bispecific antibody of paragraph 120, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18. 122. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (1) (a) a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (a) a second antigen-binding domain that specifically binds to an antigen-binding domain, wherein the method comprises administering the bispecific antibody to the individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence of SEQ ID NO: 18; (b) an anti-metabolic drug (e.g., pemetrexed), wherein in the aspect that the anti-metabolic drug is pemetrexed, the method comprises administering pemetrexed to the individual at a dose of 500 mg/ ^m2 every three weeks; and (c) A platinum-based chemotherapeutic agent (e.g., carboplatin), wherein in the aspect of the platinum-based chemotherapeutic agent being carboplatin, the method comprises administering carboplatin to the individual at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks; or (2) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering the bispecific antibody to the individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, comprising the following hypervariable regions (HVRs): an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4; and (ii) a second antigen-binding domain that specifically binds to LAG3, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising SEQ ID NO: 10 (a) an amino acid sequence of SEQ ID NO: 11, an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; (b) an anti-metabolic drug (e.g., pemetrexed), wherein in the aspect that the anti-metabolic drug is pemetrexed, the method comprises administering pemetrexed to the individual at a dose of 500 mg/ ^m2 every three weeks; and (c) a platinum-based chemotherapy agent (e.g., carboplatin), wherein in the aspect that the platinum-based chemotherapy agent is carboplatin, the method comprises administering carboplatin to the individual at a target area under the concentration-time curve (AUC) of 5 mg/mL•min every three weeks. 123. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3 and one or more chemotherapeutic agents, wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual one or more of the following dosing cycles: (1) (a) a bispecific antibody targeting PD-1 and LAG3 comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3; (a) a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence of SEQ ID NO: 18; (b) a taxane (e.g., paclitaxel), wherein in the aspect that the taxane is paclitaxel, the method comprises administering paclitaxel to the individual at a fixed dose of 600 ^mg every three weeks; and (c) A platinum-based chemotherapeutic agent (e.g., carboplatin), wherein in the aspect of the platinum-based chemotherapeutic agent being carboplatin, the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL • min every three weeks; or (2) (a) a bispecific antibody targeting PD-1 and LAG3, comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering the bispecific antibody to the individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, comprising the following HVR: HVR-H1 sequence, comprising SEQ ID NO: 1 an amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4; and (ii) a second antigen-binding domain that specifically binds to LAG3, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4. NO: 11, and HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; (b) a taxane (e.g., paclitaxel), wherein in the aspect of the taxane being paclitaxel, the method comprises administering paclitaxel to the individual at a dose of 200 mg/m ² every three weeks; and (c) a platinum-based chemotherapeutic agent (e.g., carboplatin), wherein in the aspect of the platinum-based chemotherapeutic agent being carboplatin, the method comprises administering carboplatin to the individual at a target AUC of 5 mg/mL • min every three weeks. 124. The bispecific antibody of any one of paragraphs 63 to 123, wherein the individual is human. 125. A method of treating an individual having cancer (e.g., triple negative breast cancer (TNBC)), wherein the method comprises a dosing regimen comprising concurrently administering to the individual: (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antibody binding domain that specifically binds to LAG3; and (b) one or more dosing cycles of a chemotherapy agent (e.g., a taxane, e.g., nab-paclitaxel). 126. The method of paragraph 125, wherein the individual has not previously received systemic anticancer therapy for locally advanced, unresectable or metastatic cancer (e.g., TNBC). 127. The method of paragraphs 125 or 126, wherein the subject has not been previously treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody. 128. The method of any of paragraphs 125 to 127, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 129. The method of any of paragraphs 125 to 128, wherein the length of each of the one or more dosing cycles of the bispecific antibody is about 21 days (e.g., is 21 days). 130. The method of paragraph 129, wherein the method comprises administering the bispecific antibody to the individual on or about day 1 (e.g., on day 1 ± 1 day) of each of the one or more dosing cycles. 131. The method of any of paragraphs 125 to 130, wherein the method comprises administering the bispecific antibody intravenously to the individual. 132. The method of paragraph 131, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in the first dosing cycle; and/or (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles. 133. The method of any of paragraphs 125 to 132, wherein the method comprises administering to the subject nab-paclitaxel once a week at a dose of about 100 mg/m ² (eg, once a week at a dose of 100 mg/m ² ) for three weeks followed by one week off. 134. The method of any of paragraphs 125 to 133, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg every three weeks (e.g., at a fixed dose of 600 mg every three weeks); and administering nab-paclitaxel to the subject once a week at a dose of about 100 mg/m ² (e.g., once a week at a dose of 100 mg/m ² ) for three weeks, followed by 1 week off. 135. The method of any of paragraphs 125 to 134, wherein the length of each of the one or more dosing cycles of nab-paclitaxel is about 28 days (e.g., is 28 days). 136. The method of paragraph 135, wherein the method comprises administering nab-paclitaxel to the subject on days 1, 8, and 15 of each of the one or more dosing cycles. 137. The method of any of paragraphs 125 to 136, wherein the method comprises administering nab-paclitaxel intravenously to the subject. 138. The method of paragraph 137, wherein the nab-paclitaxel is administered over about 30 minutes (e.g., administered over 30 minutes). 139. The method of any of paragraphs 125 to 138, wherein, on a day when the bispecific antibody and the chemotherapy (e.g., a taxane, e.g., nab-paclitaxel) are administered on the same day, the method comprises administering the bispecific antibody to the individual prior to the chemotherapy (e.g., a taxane, e.g., nab-paclitaxel). 140. The method of any of paragraphs 125 to 139, wherein the cancer is locally advanced, unresectable, or metastatic (e.g., the TNBC is locally advanced, unresectable, or metastatic TNBC). 141. The method of any of paragraphs 125 to 140, wherein the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases. 142. The method of any of paragraphs 125 to 141, wherein the cancer is PD-L1 positive (e.g., the TNBC is PD-L1 positive TNBC). 143. The method of paragraph 142, wherein the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) has a PD-L1 combined positivity score (CPS) of ≥ 10 as measured using the pharmDx 22C3 IHC assay. 144. The method of paragraph 142, wherein the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) has a PD-L1 tumor area positivity score (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay. 145. The method of paragraph 142, wherein the PD-L1 combined immune cell (IC) ratio of the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) is ≥ 1% as measured using the Ventana SP142 IHC assay. 146. The method of any of paragraphs 125 to 145, wherein the dosing regimen includes at least 5 or at least 10 dosing cycles of (a) the bispecific antibody and (b) a chemotherapy agent (e.g., a taxane, e.g., nab-paclitaxel). 147. The method of any of paragraphs 125 to 146, wherein the method increases progression-free survival (PFS) compared to a reference PFS. 148. The method of paragraph 147, wherein the reference PFS is the PFS of a population of individuals who have received control therapy. 149. The method of any of paragraphs 125 to 148, wherein the method increases the objective response rate (ORR) of a population of individuals treated according to the method as compared to a reference ORR. 150. The method of paragraph 149, wherein the reference ORR is the ORR of a population of individuals who have received control therapy. 151. The method of any of paragraphs 125 to 150, wherein the method increases the duration of response (DOR) as compared to a reference DOR. 152. The method of paragraph 151, wherein the reference DOR is the DOR of a population of individuals who have received control therapy. 153. The method of any of paragraphs 125 to 152, wherein the method results in an increase in OS compared to a reference overall survival (OS). 154. The method of paragraph 153, wherein the reference OS is the OS of a population of individuals who have received control therapy. 155. The method of any of paragraphs 125 to 154, wherein the method results in an increase in the PFS rate at 12 months compared to a reference PFS rate at 12 months. 156. The method of paragraph 155, wherein the reference PFS rate is the PFS rate of a population of individuals who have received control therapy. 157. The method of any of paragraphs 125 to 156, wherein the method results in an increase in the OS rate at 12 months compared to a reference OS rate at 12 months. 158. The method of paragraph 157, wherein the reference OS rate is the OS rate of a population of individuals who have received a control therapy. 159. The method of any of paragraphs 148 to 158, wherein the control therapy comprises pembrolizumab and chemotherapy (eg, a taxane, eg, nab-paclitaxel) and does not comprise a bispecific antibody. 160. The method of any one of paragraphs 125 to 159, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising an amino acid sequence of GGR; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 3; (ii) an HVR-L2 sequence comprising an amino acid sequence of RSS; and (iii) an HVR-L3 sequence comprising SEQ ID NO: 4 161. The method of paragraph 160, wherein the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. 162. The method of paragraph 161, wherein the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. 163. The method of any one of paragraphs 161 or 162, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. 164. The method of paragraph 163, wherein the Fc receptor is an Fcγ receptor. 165. The method of any one of paragraphs 160 to 164, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising SEQ ID NO: 12 166. The method of any one of paragraphs 160 to 165, wherein the first antigen-binding fragment comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14. 167. The method of any one of paragraphs 160 to 166, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the association of the first unit of the Fc domain with the second unit. 168. The method of any one of paragraphs 161 to 167, wherein the first unit of the Fc domain comprises amino acid substitutions S354C and T366W (numbered according to the Kabat EU index), and the second unit of the Fc domain comprises amino acid substitutions Y349C, T366S, and Y407V (numbered according to the Kabat EU index). 169. The method of any one of paragraphs 160 to 168, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising the second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14; and an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to Kabat EU 170. The method of paragraph 169, wherein in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. 171. The method of paragraph 170, wherein in the first Fab fragment, the variable domains VL and VH are replaced with each other. 172. The method of any one of paragraphs 45 to 47, wherein in the constant domain CL of one of the Fab fragments, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 173. A method as described in paragraph 172, wherein in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted with lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted with glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 174. The method of any one of paragraphs 169 to 173, wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. 175. The method of paragraph 174, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18. 176. A method for treating an individual with locally advanced, unresectable or metastatic cancer (e.g., locally advanced, unresectable or metastatic TNBC), wherein the method comprises a dosing regimen comprising administering to the individual concurrently: (1) (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, wherein the method comprises administering to the individual the bispecific antibody at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising SEQ ID NO: 16 an amino acid sequence of SEQ ID NO: 17; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of a chemotherapeutic agent (e.g., a taxane, e.g., albumin-bound paclitaxel), wherein in an aspect where the chemotherapeutic agent is albumin-bound paclitaxel, the method comprises administering albumin- ^bound paclitaxel to the individual once a week for three weeks, followed by one week of rest; or (2) (a) one or more dosing cycles of a bispecific antibody targeting PD-1 and LAG3, the bispecific antibody comprising a PD-1-binding protein and a PD-2-binding protein. The invention relates to a method for treating a subject with a bispecific antibody comprising: (i) a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering the bispecific antibody to a subject at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1, an HVR-H2 sequence comprising the amino acid sequence GGR, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3, an HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4; and (ii) a second antigen-binding domain that specifically binds to LAG3, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; and (b) one or more administration cycles of a chemotherapeutic agent (e.g., a taxane, e.g., albumin-bound paclitaxel), wherein in the aspect that the chemotherapeutic agent is albumin-bound paclitaxel, the method comprises administering 100 μg of the chemotherapeutic agent once a week to the patient. The subject is administered nab-paclitaxel at a dose of 1 mg/m ² for three weeks followed by one week off. 177. The method of any one of paragraphs 125 to 176, wherein the subject is a human. 178. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual having cancer (e.g., triple-negative breast cancer (TNBC)), wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3 and a chemotherapeutic agent (e.g., a taxane, e.g., nab-paclitaxel), wherein the bispecific antibody comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual concurrently: (a) one or more dosing cycles of the bispecific antibody; and (b) the chemotherapeutic agent (e.g., a taxane, e.g., nab-paclitaxel) 179. The bispecific antibody of paragraph 178, wherein the individual has not previously received systemic anticancer therapy for locally advanced, unresectable or metastatic cancer (e.g., TNBC). 180. The bispecific antibody of paragraphs 178 or 179, wherein the individual has not previously been treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody. 181. The bispecific antibody of any of paragraphs 178 to 180, wherein the method comprises administering the bispecific antibody to the individual at a fixed dose of about 600 mg (e.g., at a fixed dose of 600 mg) every three weeks. 182. The bispecific antibody of any of paragraphs 178 to 181, wherein the length of each of the one or more dosing cycles of the bispecific antibody is about 21 days (e.g., is 21 days). 183. The bispecific antibody of paragraph 182, wherein the method comprises administering the bispecific antibody to the individual on or about day 1 (e.g., on day 1 ± 1 day) of each of the one or more dosing cycles. 184. The bispecific antibody of any of paragraphs 178 to 183, wherein the method comprises administering the bispecific antibody intravenously to the individual. 185. A bispecific antibody as described in paragraph 184, wherein: (a) the bispecific antibody is administered over 60 (± 15) minutes in a first dosing cycle; and/or (b) the method comprises one or more additional dosing cycles, and the bispecific antibody is administered over 30 (± 10) minutes in the one or more additional dosing cycles. 186. The bispecific antibody of any of paragraphs 178 to 185, wherein the method comprises administering to the individual nab-paclitaxel once a week at a dose of about 100 mg/m ² (e.g., once a week at a dose of 100 mg/m ² ) for three weeks followed by one week off. 187. The bispecific antibody of any one of paragraphs 178 to 186, wherein the method comprises administering the bispecific antibody to the subject at a fixed dose of about 600 mg every three weeks (e.g., at a fixed dose of 600 mg every three weeks); and administering nab-paclitaxel to the subject once a week at a dose of about 100 mg/m ² (e.g., once a week at a dose of 100 mg/m ² ) for three weeks, followed by 1 week off. 188. The bispecific antibody of any one of paragraphs 178 to 186, wherein the length of each of the one or more dosing cycles of nab-paclitaxel is about 28 days (e.g., is 28 days). 189. The bispecific antibody of paragraph 188, wherein the method comprises administering to the subject nab-paclitaxel on days 1, 8, and 15 of each of the one or more dosing cycles. 190. The bispecific antibody of any of paragraphs 178 to 189, wherein the method comprises administering to the subject nab-paclitaxel intravenously. 191. The bispecific antibody of paragraph 190, wherein the nab-paclitaxel is to be administered over about 30 minutes (e.g., administered over 30 minutes). 192. The bispecific antibody of any one of paragraphs 178 to 191, wherein the bispecific antibody and the chemotherapy (e.g., taxane, e.g., nab-paclitaxel) are to be administered on the same day, the method comprising administering the bispecific antibody to the individual prior to the chemotherapy (e.g., taxane, e.g., nab-paclitaxel). 193. The bispecific antibody of any one of paragraphs 178 to 192, wherein the cancer is locally advanced, unresectable or metastatic (e.g., TNBC is locally advanced, unresectable or metastatic TNBC). 194. The bispecific antibody of any of paragraphs 178 to 193, wherein the individual does not have any symptomatic, untreated, or actively progressing central nervous system (CNS) metastases. 195. The bispecific antibody of any of paragraphs 178 to 194, wherein the cancer is PD-L1 positive (e.g., the TNBC is PD-L1 positive TNBC). 196. The bispecific antibody of paragraph 195, wherein the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) has a PD-L1 combined positivity score (CPS) of ≥10 as measured using the pharmDx 22C3 IHC assay. 197. The bispecific antibody of paragraph 195, wherein the PD-L1 tumor area positivity score (TAP) of the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) is ≥ 5% as measured using the Ventana SP263 IHC assay. 198. The bispecific antibody of paragraph 195, wherein the PD-L1 combined immune cell (IC) ratio of the PD-L1 positive cancer (e.g., PD-L1 positive TNBC) is ≥ 1% as measured using the Ventana SP142 IHC assay. 199. The bispecific antibody of any of paragraphs 178 to 198, wherein the dosing regimen comprises at least 5 or at least 10 dosing cycles of (a) the bispecific antibody and (b) a chemotherapy agent (e.g., a taxane, e.g., nab-paclitaxel). 200. The bispecific antibody of any of paragraphs 178 to 199, wherein the method results in an increase in progression-free survival (PFS) compared to a reference PFS. 201. The bispecific antibody of paragraph 200, wherein the reference PFS is the PFS of a population of individuals who have received control therapy. 202. The bispecific antibody of any of paragraphs 178 to 201, wherein the method results in an increase in the objective response rate (ORR) of a population of individuals treated according to the method compared to a reference ORR. 203. The bispecific antibody of paragraph 202, wherein the reference ORR is the ORR of a population of individuals who have received control therapy. 204. The bispecific antibody of any of paragraphs 178 to 203, wherein the method results in an increase in the duration of response (DOR) compared to a reference DOR. 205. The bispecific antibody of paragraph 204, wherein the reference DOR is the DOR of a population of individuals who have received control therapy. 206. The bispecific antibody of any of paragraphs 178 to 205, wherein the method results in an increase in OS compared to a reference overall survival (OS). 207. The bispecific antibody of paragraph 206, wherein the reference OS is the OS of a population of individuals who have received control therapy. 208. The bispecific antibody of any of paragraphs 178 to 207, wherein the method results in an increase in the PFS rate at 12 months compared to a reference PFS rate at 12 months. 209. The bispecific antibody of paragraph 208, wherein the reference PFS rate is the PFS rate of a population of individuals who have received control therapy. 210. The bispecific antibody of any of paragraphs 178 to 209, wherein the method results in an increase in OS rate at 12 months compared to a reference OS rate at 12 months. 211. The bispecific antibody of paragraph 210, wherein the reference OS rate is the OS rate in a population of individuals who have received a control therapy. 212. The bispecific antibody of any of paragraphs 201 to 211, wherein the control therapy comprises pembrolizumab and a chemotherapy agent (e.g., a taxane, e.g., nab-paclitaxel) and does not comprise the bispecific antibody. 213. The bispecific antibody of any one of paragraphs 178 to 212, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising the amino acid sequence GGR; and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3; (ii) an HVR-L2 sequence comprising the amino acid sequence RSS; and (iii) an HVR-L3 sequence comprising the amino acid sequence RSS. A sequence comprising the amino acid sequence of SEQ ID NO: 4. 214. The bispecific antibody of paragraph 213, wherein the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. 215. The bispecific antibody of paragraph 214, wherein the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. 216. The bispecific antibody of any one of paragraphs 214 or 215, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. 217. The bispecific antibody of paragraph 216, wherein the Fc receptor is an Fcγ receptor. 218. The bispecific antibody of any one of paragraphs 213 to 217, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10, (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11, and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 12. 219. The bispecific antibody of any one of paragraphs 213 to 218, wherein the first antigen-binding fragment comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14. 220. The bispecific antibody of any one of paragraphs 213 to 219, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to the Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the association of the first unit and the second unit of the Fc domain. 221. The bispecific antibody of any one of paragraphs 214 to 220, wherein the first Fc domain unit comprises the amino acid substitutions S354C and T366W (numbered according to the Kabat EU index), and the second Fc domain unit comprises the amino acid substitutions Y349C, T366S, and Y407V (numbered according to the Kabat EU index). 222. The bispecific antibody of any one of paragraphs 213 to 221, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 5, and a VL domain comprising the amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising the second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 13, and a VL domain comprising the amino acid sequence of SEQ ID NO: 14; and an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (According to the Kabat EU index numbering). 223. The bispecific antibody of paragraph 222, wherein in one of the Fab fragments of the bispecific antibody targeting PD-1 and LAG3, the variable domains VL and VH are replaced with each other, so that the VH domain is part of the light chain and the VL domain is part of the heavy chain. 224. The bispecific antibody of paragraph 223, wherein in the first Fab fragment, the variable domains VL and VH are replaced with each other. 225. The bispecific antibody of any one of paragraphs 222 to 224, wherein in the constant domain CL of one of said Fab fragments, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 226. A bispecific antibody as described in paragraph 225, wherein in the constant domain CL of the second Fab fragment, the amino acid at position 124 is independently substituted by lysine (K), arginine (R) or histidine (H) (according to the Kabat EU index numbering), and in the constant domain CH1, the amino acids at positions 147 and 213 are independently substituted by glutamine (E) or aspartic acid (D) (according to the Kabat EU index numbering). 227. The bispecific antibody of any one of paragraphs 213 to 226, wherein the bispecific antibody comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. 228. The bispecific antibody of paragraph 227, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18. 229. A bispecific antibody targeting PD-1 and LAG3 for use in a method of treating an individual with locally advanced, unresectable or metastatic cancer (e.g., locally advanced, unresectable or metastatic TNBC), wherein the method comprises a dosing regimen comprising the bispecific antibody targeting PD-1 and LAG3 and albumin-bound paclitaxel, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, and wherein the dosing regimen comprises administering to the individual concurrently: (1) (a) 600 mg every three weeks One or more dosing cycles of a fixed-dose bispecific antibody, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising the amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel, wherein the method comprises administering albumin-bound paclitaxel to the individual once a week at a dose of 100 mg/ ^m2 for three weeks, followed by 1 week of rest; or (2) (a) targeting PD-1 and LAG3 One or more dosing cycles of a bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, wherein the method comprises administering the bispecific antibody to the individual at a fixed dose of 600 mg every three weeks, and wherein the bispecific antibody comprises: (i) a first antigen-binding domain that specifically binds to PD-1, which comprises the following HVRs: HVR-H1 sequence, which comprises the amino acid sequence of SEQ ID NO: 1, HVR-H2 sequence, which comprises the amino acid sequence GGR, HVR-H3 sequence, which comprises the amino acid sequence of SEQ ID NO: 2, HVR-L1 sequence, which comprises the amino acid sequence of SEQ ID NO: 3, HVR-L2 sequence comprising the amino acid sequence RSS, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 4; and (ii) a second antigen-binding domain that specifically binds to LAG3, comprising the following HVRs: an HVR-H1 sequence comprising the amino acid sequence of SEQ ID NO: 7, an HVR-H2 sequence comprising the amino acid sequence of SEQ ID NO: 8, an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 9, an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L2 sequence comprising the amino acid sequence of SEQ ID NO: 11, and an HVR-L3 sequence comprising the amino acid sequence of SEQ ID NO: 12; and (b) a chemotherapeutic agent (e.g., a taxane, e.g., albumin-bound paclitaxel) wherein the chemotherapeutic agent is nab-paclitaxel, the method comprises administering nab-paclitaxel to the individual once a week for three weeks followed by one week off. 230. The bispecific antibody of any one of paragraphs 178 to ²²⁹ , wherein the individual is a human.

Although the above invention has been described in detail by way of illustrations and examples for the purpose of clear understanding, such descriptions and examples should not be construed as limiting the scope of the invention.

FIG1 is a flow chart showing the design of the BO44178 Phase II clinical trial of RO7247669 plus platinum- based chemotherapy compared with pembrolizumab plus platinum-based chemotherapy in patients with previously untreated locally advanced or metastatic non-small cell lung cancer (NSCLC). ADA = antidrug antibodies; DOR = duration of response; ECOG PS = Eastern Cooperative oncology performance status; IMC = internal monitoring committee; Inv = investigator; NSCLC = non-small-cell lung cancer; NSQ = nonsquamous; ORR = objective response rate; OS = overall survival; PD = progressive disease; PD-L1 = planned death-ligand 1; PFS = progression-free survival; PK = pharmacokinetics; PRO = patient-reported outcome; R = randomized; SQ = squamous. - Figure 2 is a flow diagram showing the design of the CO44194 Phase II clinical trial of RO7247669 in combination with nab-paclitaxel versus pembrolizumab in combination with nab-paclitaxel in patients with previously untreated, PD-L1-positive, locally advanced unresectable or metastatic triple-negative breast cancer (TNBC). 1L = first-line; CPS = combined positivity score; eTNBC = early triple-negative breast cancer; IMC = internal monitoring committee; PD-L1 = planned death-ligand 1; Q3W = every 3 weeks; R = randomized; TAP = tumor area positivity. ^aSafety run-in: Approximately 6 weeks after the 12th participant reaches the first study treatment. FIG3A is a schematic diagram showing the design of the first year of the CO44194 Phase II clinical trial. ADA = anti-drug antibodies; PK = pharmacokinetic. FIG3B is a schematic diagram showing the design of the CO44194 Phase II clinical trial after the second year. FIG3C is a schematic diagram showing the design of the follow-up period of the CO44194 Phase II clinical trial.

TW202436339A_113103499_SEQL.xml

Claims (91)

A use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals suffering from non-squamous non-small cell lung cancer (NSCLC), wherein the bispecific antibody is formulated for administration simultaneously or separately in combination with (1) pemetrexed and (2) carboplatin, and wherein the treatment comprises administering to the individual or group of individuals a dosing regimen comprising one or more dosing cycles of the medicament, pemetrexed, and carboplatin. The use of claim 1, wherein the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks. The use of claim 1 or 2, wherein pemetrexed is formulated for administration at a dose of 500 mg/m ² every three weeks. The use of claim 1 or 2, wherein the calcitonin is formulated for administration every three weeks at a target area under the concentration-time curve (AUC) of 5 mg/mL • min. The use of claim 1, wherein the length of each of the one or more dosing cycles is 21 days, and wherein the treatment comprises administering the bispecific antibody targeting PD-1 and LAG3, pemetrexed, and carboplatin on day 1 of each of the one or more dosing cycles. The use of claim 1, wherein the bispecific antibody targeting PD-1 and LAG3, the pemetrexed and the carboplatin are formulated for intravenous administration. The use of claim 1, wherein the treatment comprises: (a) administering the bispecific antibody targeting PD-1 and LAG3 over a period of 60 (± 15) minutes in a first dosing cycle and over a period of 30 (± 10) minutes in one or more additional dosing cycles; (b) administering pemetrexed over a period of 10 minutes; and/or (c) administering carboplatin over a period of 30 to 60 minutes. The use of claim 1, wherein the dosing regimen comprises four dosing cycles. The use of claim 1, wherein the dosing regimen further comprises one or more additional dosing cycles of (a) the bispecific antibody targeting PD-1 and LAG3 and (b) pemetrexed. The use of claim 9, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody targeting PD-1 and LAG3 and pemetrexed. A use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals suffering from squamous NSCLC, wherein the bispecific antibody is formulated for administration simultaneously or separately in combination with (1) paclitaxel and (2) carboplatin, and wherein the treatment comprises administering to the individual or group of individuals a dosing regimen comprising one or more dosing cycles of the medicament, paclitaxel, and carboplatin. The use of claim 11, wherein the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks. The use of claim 11 or 12, wherein the paclitaxel is formulated for administration at a dose of 200 mg/m ² every three weeks. The use of claim 11 or 12, wherein the platinum is formulated for administration every three weeks at a target AUC of 5 mg/mL • min. The use of claim 11, wherein the length of each of the one or more dosing cycles is 21 days, and wherein the treatment comprises administering the bispecific antibody targeting PD-1 and LAG3, the paclitaxel, and the carboplatin on day 1 of each of the one or more dosing cycles. The use of claim 11, wherein the bispecific antibody targeting PD-1 and LAG3, the paclitaxel and the carboplatin are formulated for intravenous administration. The use of claim 11, wherein the treatment comprises: (a) administering the bispecific antibody over a period of 60 (± 15) minutes in a first dosing cycle and over a period of 30 (± 10) minutes in one or more additional dosing cycles; (b) administering paclitaxel over a period of 3 hours; and/or (c) administering carboplatin over a period of 30 to 60 minutes. The use of claim 11, wherein the dosing regimen comprises four dosing cycles. The use of claim 18, wherein the dosing regimen further comprises one or more additional dosing cycles of the bispecific antibody targeting PD-1 and LAG3. The use of claim 19, wherein the dosing regimen comprises at least 5 or at least 10 additional dosing cycles of the bispecific antibody targeting PD-1 and LAG3. The use of claim 11, wherein the subject is premedicated with oral or IV steroids and antihistamines prior to administration of the paclitaxel. For the use of claim 1 or 11, wherein the NSCLC is: (a) Stage IIIB/IIIC NSCLC; or (b) Stage IV NSCLC. The use of claim 1 or 11, wherein the individual has not received prior systemic treatment for metastatic NSCLC. The use according to claim 1 or 11, wherein the individual has not been previously treated with immune checkpoint blockade therapy. The use of claim 1 or 11, wherein the individual has not been previously treated with an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) agent, an anti-T-cell immunoreceptor with Ig and tyrosine-based inhibitory motif domain (TIGIT) agent, an anti-PD-1 therapeutic antibody, an anti-PD-L1 therapeutic antibody, or an anti-LAG3 agent. The use of claim 1 or 11, wherein the individual has not been previously treated with a CD137 agonist. The use of claim 1 or 11, wherein the individual does not have any symptomatic, untreated or actively progressing central nervous system (CNS) metastases. The use of claim 1 or 11, wherein the individual has no known mutation in the epidermal growth factor receptor (EGFR) gene or the anaplastic lymphoma kinase (ALK) fusion oncogene. The use of claim 1 or 11, wherein the individual has an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1. The use of claim 1 or 11, wherein the NSCLC tumor sample from the individual has a PD-L1 tumor proportion score (TPS) or PD-L1 expression on tumor cells (TC) of <1%. The use of claim 1 or 11, wherein the tumor sample of the NSCLC from the individual has a PD-L1 TPS or an expression amount of PD-L1 on TC between 1% and 49%. The use of claim 1 or 11, wherein the tumor sample of the NSCLC from the individual has a PD-L1 TPS or PD-L1 expression on TC of ≥50%. The use of claim 1 or 11, wherein the method increases progression-free survival (PFS) compared to a reference PFS. The use of claim 33, wherein the reference PFS is the PFS of a population of individuals who have received a control therapy. The use of claim 1 or 11, wherein the method results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. The use of claim 35, wherein the reference ORR is the ORR for a population of individuals who have received control therapy. The use of claim 1 or 11, wherein the treatment results in an increase in overall survival (OS) compared to a reference OS. The use of claim 37, wherein the reference OS is the OS of a population of individuals who have received a control therapy. The use of claim 1 or 11, wherein the treatment results in an increase in the duration of response (DOR) as compared to a reference DOR. For use as in claim 39, wherein the reference DOR is the DOR for the population of individuals who received the control therapy. The use of claim 1 or 11, wherein: (a) the individual has non-squamous NSCLC and the control therapy comprises pembrolizumab, pemetrexed, and carboplatin and does not comprise the bispecific antibody; or (b) the individual has squamous NSCLC and the control therapy comprises pembrolizumab, paclitaxel, and carboplatin and does not comprise the bispecific antibody. Use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals suffering from triple-negative breast cancer (TNBC), wherein the bispecific antibody is formulated for concurrent administration in combination with nab-paclitaxel, and wherein the treatment comprises administering to the individual or group of individuals a dosing regimen comprising one or more dosing cycles of the medicament and nab-paclitaxel. The use of claim 42, wherein the individual has not previously received systemic anticancer therapy for locally advanced, unresectable or metastatic TNBC. The use of claim 42, wherein the individual has not been previously treated with anti-LAG3 therapy, a CD137 agonist, or an anti-CTLA therapeutic antibody. The use of claim 42, wherein the bispecific antibody targeting PD-1 and LAG3 is formulated for administration at a fixed dose of 600 mg every three weeks. The use of claim 42, wherein the length of each of the one or more dosing cycles of the bispecific antibody is 21 days. The use of claim 46, wherein the bispecific antibody targeting PD-1 and LAG3 is administered on day 1 of each of the one or more dosing cycles. The use of claim 42, wherein the treatment comprises intravenous administration of the bispecific antibody targeting PD-1 and LAG3. The use of claim 42, wherein the treatment comprises administering the bispecific antibody targeting PD-1 and LAG3 over a period of 60 (± 15) minutes in a first dosing cycle; and/or administering the bispecific antibody targeting PD-1 and LAG3 over a period of 30 (± 10) minutes in one or more additional dosing cycles. The use of claim 42, wherein the treatment comprises administering nab-paclitaxel at a dose of 100 mg/m ² once a week for three weeks, followed by one week off. The use of claim 42, wherein the treatment comprises administering the bispecific antibody targeting PD-1 and LAG3 at a fixed dose of 600 mg every three weeks and administering albumin-bound paclitaxel at a dose of 100 mg/ ^m2 once a week for three weeks, followed by 1 week of rest. The use of claim 42, wherein the length of each of the one or more dosing cycles of nab-paclitaxel is 28 days. The use of claim 42, wherein the treatment comprises administering nab-paclitaxel on days 1, 8, and 15 of each of the one or more dosing cycles. The use of claim 42, wherein the treatment comprises intravenously administering albumin-bound paclitaxel. The use of claim 54, wherein the albumin-bound paclitaxel is administered over a period of 30 minutes. The use of claim 42, wherein, on a day when the bispecific antibody and the nab-paclitaxel are administered on the same day, the treatment comprises administering the bispecific antibody to the individual prior to the nab-paclitaxel. The use as in claim 42, wherein the TNBC is locally advanced, unresectable or metastatic TNBC. The use of claim 42, wherein the individual does not have any symptomatic, untreated, or active CNS metastases. The use of claim 42, wherein the TNBC is PD-L1-positive TNBC. The use of claim 59, wherein the PD-L1 positive TNBC has a PD-L1 composite positivity score (CPS) ≥ 10 as measured using the pharmDx 22C3 IHC assay. The use of claim 59, wherein the PD-L1 positive TNBC has a PD-L1 tumor area positivity (TAP) of ≥ 5% as measured using the Ventana SP263 IHC assay. The use of claim 59, wherein the PD-L1 positive TNBC has a PD-L1 positive immune cell (IC) fraction of ≥ 1% as measured using a Ventana SP142 IHC assay. The use of claim 42, wherein the dosing regimen comprises at least 5 or at least 10 dosing cycles of (a) the bispecific antibody targeting PD-1 and LAG3 and (b) nab-paclitaxel. The use of claim 42, wherein the treatment results in an increase in progression-free survival (PFS) as compared to a reference PFS. The use of claim 64, wherein the reference PFS is the PFS of a population of individuals who have received a control therapy. The use of claim 42, wherein the treatment results in an increase in the objective response rate (ORR) in a population of individuals treated according to the method as compared to a reference ORR. The use of claim 66, wherein the reference ORR is the ORR for a population of individuals who have received control therapy. The use of claim 42, wherein the treatment results in an increase in the duration of response (DOR) as compared to a reference DOR. For use as described in claim 68, wherein the reference DOR is the DOR for the population of individuals who received the control therapy. The use of claim 42, wherein the method results in an increase in overall survival (OS) compared to a reference OS. The use of claim 70, wherein the reference OS is the OS of a population of individuals who have received a control therapy. The use of claim 42, wherein the treatment results in an increase in the PFS rate at 12 months compared to a reference PFS rate at 12 months. For the purpose of claim 72, wherein the reference PFS rate is the PFS rate for a population of individuals who have received a control therapy. The use of claim 42, wherein the treatment results in an increase in OS rate at 12 months compared to a reference OS rate at 12 months. The use of claim 74, wherein the reference OS rate is the OS rate in a population of individuals who have received a control therapy. The use of claim 42, wherein the control therapy comprises pembrolizumab and nab-paclitaxel and does not comprise the bispecific antibody targeting PD-1 and LAG3. The use of claim 1 or 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first antigen-binding domain that specifically binds to PD-1, the first antigen-binding domain comprising: a heavy chain variable (VH) domain comprising: (i) a highly variable region H1 (HVR-H1) sequence comprising the amino acid sequence of SEQ ID NO: 1; (ii) an HVR-H2 sequence comprising the amino acid sequence GGR; and (iii) an HVR-H3 sequence comprising the amino acid sequence of SEQ ID NO: 2; and a light chain variable (VL) domain comprising: (i) an HVR-L1 sequence comprising the amino acid sequence of SEQ ID NO: 3; (ii) an HVR-L2 sequence comprising the amino acid sequence RSS; and (iii) HVR-L3 sequence, which comprises the amino acid sequence of SEQ ID NO: 4. The use of claim 1, 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises an Fc domain, and the Fc domain is IgG. The use of claim 1, 11 or 42, wherein the IgG Fc domain is an IgG1 Fc domain or an IgG4 Fc domain. The use of claim 79, wherein the Fc domain comprises one or more amino acid substitutions that reduce binding to an Fc receptor. The use of claim 80, wherein the Fc receptor is an Fcγ receptor. The use of claim 1 or 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising: a VH domain comprising: (i) an HVR-H1 sequence comprising an amino acid sequence of SEQ ID NO: 7; (ii) an HVR-H2 sequence comprising an amino acid sequence of SEQ ID NO: 8; and (iii) an HVR-H3 sequence comprising an amino acid sequence of SEQ ID NO: 9; and a VL domain comprising: (i) an HVR-L1 sequence comprising an amino acid sequence of SEQ ID NO: 10; (ii) an HVR-L2 sequence comprising an amino acid sequence of SEQ ID NO: 11; and (iii) an HVR-L3 sequence comprising an amino acid sequence of SEQ ID NO: 9. A sequence comprising the amino acid sequence of SEQ ID NO: 12. The use of claim 1 or 11 or 42, wherein the first antigen-binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 5 and a VL domain comprising the amino acid sequence of SEQ ID NO: 6, and the second antigen-binding domain comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 13 and a VL domain comprising the amino acid sequence of SEQ ID NO: 14. The use of claim 1, 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: (a) an Fc domain of human IgG1 subclass having amino acid mutations L234A, L235A and P329G (numbered according to Kabat EU index); and/or (b) the Fc domain comprising a modification that promotes the binding of the first unit and the second unit of the Fc domain. The use of claim 1 or 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises a first Fab fragment comprising the first antigen-binding domain that specifically binds to PD1, the first antigen-binding domain comprising a VH domain comprising an amino acid sequence of SEQ ID NO: 5 and a VL domain comprising an amino acid sequence of SEQ ID NO: 6; and a second Fab fragment comprising the second antigen-binding domain that specifically binds to LAG3, the second antigen-binding domain comprising a VH domain comprising an amino acid sequence of SEQ ID NO: 13 and a VL domain comprising an amino acid sequence of SEQ ID NO: 14; and an Fc domain of the human IgG1 subclass having amino acid mutations L234A, L235A and P329G (according to Kabat EU index number). The use of claim 1 or 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: a first heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence having at least 95% sequence identity with the sequence of SEQ ID NO: 18. The use of claim 1 or 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 comprises: a first heavy chain comprising an amino acid sequence of SEQ ID NO: 15; a first light chain comprising an amino acid sequence of SEQ ID NO: 16; a second heavy chain comprising an amino acid sequence of SEQ ID NO: 17; and a second light chain comprising an amino acid sequence of SEQ ID NO: 18. The use of claim 1, 11 or 42, wherein the bispecific antibody targeting PD-1 and LAG3 is tobemstomig. A use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals with non-squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the treatment comprises administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) the bispecific antibody targeting PD-1 and LAG3, administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) pemetrexed administered at a dose of 500 mg/m ² every three weeks; and (c) carboplatin administered at a target area under the concentration-time curve (AUC) of 5 mg/mL • min every three weeks. A use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen binding domain that specifically binds to PD-1 and a second antigen binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals with squamous stage IIIB/IIIC NSCLC or stage IV NSCLC, wherein the treatment comprises administering to the individual a dosing regimen comprising one or more dosing cycles of: (a) the bispecific antibody targeting PD-1 and LAG3, administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising SEQ ID NO: 15 an amino acid sequence of SEQ ID NO: 16; a first light chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; (b) paclitaxel administered at a dose of 200 mg/m ² every three weeks; and (c) carboplatin administered at a target AUC of 5 mg/mL • min every three weeks. A use of a bispecific antibody targeting PD-1 and LAG3 in the manufacture of a medicament, the bispecific antibody comprising a first antigen-binding domain that specifically binds to PD-1 and a second antigen-binding domain that specifically binds to LAG3, the medicament being used to treat an individual or a group of individuals with locally advanced, unresectable or metastatic TNBC, wherein the treatment comprises a dosing regimen comprising administering to the individual concurrently: (a) one or more dosing cycles of the bispecific antibody targeting PD-1 and LAG3 administered at a fixed dose of 600 mg every three weeks, wherein the bispecific antibody comprises: a first heavy chain comprising the amino acid sequence of SEQ ID NO: 15; a first light chain comprising SEQ ID NO: 2 NO: 16; a second heavy chain comprising the amino acid sequence of SEQ ID NO: 17; and a second light chain comprising the amino acid sequence of SEQ ID NO: 18; and (b) one or more dosing cycles of albumin-bound paclitaxel administered once weekly at a dose of 100 mg/m ² for three weeks, followed by one week off.