RU2849612C2 - Conjugate antibody preparation and its use - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a binding agent containing an amino acid unit (RL), wherein RL contains: -Val-Ala-Q-Gly-Phe-Gly-, where Q represents a bond, a single amino acid or a dipeptide, and its antibody-drug conjugate (ADC).

EFFECT: ADC drugs can release low-molecular-weight drugs more effectively in tumour cells by optimising the polypeptide sequence in the binding agent.

10 cl, 15 dwg, 3 tbl, 4 ex

Description

Field to which the invention relates

The present patent application relates to the field of biological medicine, in particular to a binding agent and its antibody-drug conjugate (ADC).

General information about the invention

In cancer treatment, many cytotoxic drug molecules are unsuitable due to their inability to selectively poison cancer cells. Therefore, the antibody-drug conjugate (ADC) has been developed as a new therapeutic option for cancer. Generally, an ADC consists of three parts: an antibody, a binding agent, and a drug, with the antibodies linked by the binding agent and the drug. The mechanism of action of the antibody-drug complex is to first recognize a specific or overexpressed antigen on the tumor cell due to the antibody's high recognition affinity for the corresponding antigen and then bind to it. When the resulting conjugate enters the tumor cell via endocytosis, the bound cytotoxic drug molecules also enter the tumor. In tumor cells, the antibodies or binding agents are degraded to release the cytotoxic drug molecules to kill the target tumor cells, thereby achieving selective poisoning.

Binding agents used in ADCs must meet several requirements: During circulation in human plasma, binding agents must be stable to prevent premature drug release; during tumor cell penetration, cleavable binding agents can be cleaved under certain conditions to release the drug, whereas non-cleavable binding agents are released in an active form containing the drug, the binding agent, and amino acid residues derived from protease degradation ligands.

Since the primary mechanism of action of lysable antibody-drug conjugates is that antibodies target specific cell surface antigens, they enter the cellular endoplasmic reticulum system via endocytosis after binding and release small-molecule drugs via protease cleavage in the endosome or lysosome to target and kill tumor cells. Due to the heterogeneity of solid tumors, target antigens are not expressed or expressed at low levels in some tumors. Therefore, small molecules with transmembrane activity can kill adjacent tumor cells upon release, a phenomenon known as the bystander effect.

Cathepsin B is a protease found in endosomes or lysosomes, one of whose key components is cathepsin B. This protease belongs to the family of cysteine proteases and hydrolases. It is homologous to papain (trypsin), and its optimal pH is acidic. This enzyme is involved in the dissolution of the extracellular matrix, thereby influencing tumor infiltration and metastasis. Therefore, it is highly expressed in various tumor cells, such as gastric cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, and kidney cancer. ADCs entering tumor cells are rapidly lysed by large amounts of cathepsin B, rapidly releasing small molecules that destroy cancer cells.

Suboptimal specificity can lead to decreased ADC efficacy, reduced immunological specificity of conjugates, and increased toxicity due to nonspecific drug release from conjugates. Therefore, new technologies for binding agents and conjugates available for targeted therapy are needed.

Brief description of the invention

This patent application proposes a binding agent comprising a spacer (an amino acid unit) capable of independent chemical or biological degradation. The amino acid unit comprises a GFG substrate component, preferred by cathepsin B, and a VA sequence added to the N-terminus of the GFG fragment to enhance binding to the active lysis region of cathepsin B. Considering the influence of polypeptide B length on enzyme substrate activity, one S and two S can be added between VA and GF, respectively, to form three polypeptide sequences: VAGFG, VASGFG, and VASSGFG. Consequently, ADCs can more effectively release small molecule drugs in tumor cells with optimized polypeptide sequences.

This application also discloses a binding agent drug, including the binding agent, conjugates and their use.

On the one hand, the present patent application relates to a linking agent, wherein the linking agent comprises an amino acid unit (RL), wherein RL comprises the following: -Val-Ala-Q-Gly-Phe-Gly-, where Q indicates a linkage, one amino acid or a dipeptide.

In some embodiments, RL may be lysed by enzymes.

In some embodiments, where RL may be lysed by cathepsin B.

In some embodiments, where Q indicates a connection.

In some embodiments, where Q indicates -Ser- or -Ser-Ser-.

In some embodiments, where RL combines -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG), or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG).

In some embodiments, the binding agent is selected from the following sequences: -Z-A-S*-RL- and -Z-A-S*-RL-Y-; where Z denotes a subclone and A denotes a linkage or co-unit, where S* denotes a linkage or separating agent, and Y denotes a spacer subunit.

In some embodiments, wherein Z comprises the following structure: -(succinimide-3-yl-N)-, -CH ₂ -C(=O)-NH-, or -C (=O)-;

where (succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

In some embodiments, where A is selected from the following structure:

-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ heteroalkylene-, -C ₃ -C ₈ carbocycloalkyl-, -O-(C ₁ -C ₈ alkylene)-, -arylene-, -C ₁ -C ₁₀ alkylene-arylene-, -arylene-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-, -C ₃ -C ₈ heterocyclyl-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ heteroalkylene-C(=O)-, -C ₃ -C ₈ carbocycloalkyl-C(=O)-, -O-(C ₁ -C ₈ alkylene)-C(=O)-, -arylene-C(=O)-, -C ₁ -C ₁₀ alkylene-arylene-C(=O)-, -arylene-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-C(=O)-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₃ -C ₈ heterocyclyl-C(=O)-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-C(=O)-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ heteroalkylene-NH-, -C ₃ -C ₈ carbocycloalkyl-NH-, -O-(C ₁ -C ₈ alkylene)-NH-, -arylene-NH-, -C ₁ -C ₁₀ alkylene-arylene-NH-, -arylene-C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-NH-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-NH-, -C ₃ -C ₈ heterocyclyl-NH-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-NH-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ alkylene-S-, -C ₁ -C ₁₀ heteroalkylene-S-, -C ₃ -C ₈ carbocycloalkyl-S-, -O-(C ₁ -C ₈ alkylene)-S-, -arylene-S-, -C ₁ -C ₁₀ alkylene-arylene-S-, -arylene-C ₁ -C ₁₀ alkylene-S-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-S-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-S-, -C ₃ -C ₈ heterocyclyl -S-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-S- or -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-S-;

wherein A is further replaced by a basic unit (BU) that contains -( _CH2 ) _xNH2 , _- ( _CH2 ) _xNHRa ^, or -( _CH2 ) _xNRa2 where x is any integer from 1 to ₄ ; further, each R ^a is independently selected from C1 _{- C6} alkyl and C1 _{- C6} haloalkyl, or two R ^a groups are combined ^with nitrogen attached thereto to form a 4-6-membered heteroaryl or azetidinyl, pyrrolidyl, or piperidyl group.

In some embodiments, where A is selected from the following structure: -(C ₁ -C ₁₀ ) alkylene-C(=O)-, wherein the alkylene portion of A is further replaced by a basic unit (BU).

In some embodiments, the binding agent may comprise the following structure:

, where a specifies an integer from 1 to 8.

In some embodiments, where a indicates a number from 2 to 5.

In some embodiments, where Y comprises the structure shown in -NH-(CH ₂ )n ¹ -La-Lb-Lc-, where La is -C(=O)-NH-, -NR ¹ -(CH ₂ )n ² -, -O-, or a single bond, and R ¹ is H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkylene carboxyl, or C ₁ -C ₆ alkylene hydroxyl.

Lb indicates ^-CR2 ( ^-R3 )- or one bond, where ^R2 and ^R3 independently indicate H, _-NH2 , _C1 - _C6 alkylene, _C1 - _C6 alkylene-amino, _C1 - _C6 alkylene- ^carboxyl , or C1- _C6 alkylene-hydroxyl, and _R2 and ^R3 are not simultaneously _-NH2 .

n ¹ indicates an integer from 0 to 6;

Lc indicates -CH ₂ - or -C(=O)-.

In some embodiments, where Y comprises NH-(CH ₂ )n ¹ -O-(CH ₂ )n ² -C(=O)-, where n ¹ or n ² is independently any integer from 0 to 5.

In some embodiments, where Y combines -NH-CH ₂ -C(=O)-, -NH-CH ₂ CH ₂ -C(=O)-, -NH-(CH ₂ ) ₃ -C(=O)-, -NH-(CH ₂ ) ₄ -C(=O)-, -NH-(CH ₂ ) ₅ -C(=O)-, -NH-CH ₂ -O-CH ₂ -C(=O)-, or -NH-(CH ₂ ) ₂ -O-CH ₂ -C(=O)-.

In some embodiments, where S* indicates a bond and the coupling agent indicates -Z-A-RL- or -Z-A-RL-Y-.

In some embodiments, where S* indicates a separating agent and the coupling agent indicates -Z-A-S*-RL- or -Z-A-S*-RL-Y-.

In some embodiments, where S* incorporates a PEG unit.

In some embodiments, S* combines the following formula:

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ C(=O)-;

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ C(=O)NH-(CH ₂ CH ₂ O)-CH ₂ CH ₂ C(=O)-; or

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ NH-(CH ₂ CH ₂ O)-CH ₂ CH ₂ C(=O)-;

where the left and right sides of S* are associated with A and RL respectively, and b is any integer from 2 to 20.

In some embodiments, where b indicates 2, 4, 8, or 12.

In some embodiments, the binding agent is selected from the following structure:

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-C(=O)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

Where,

(succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG), or Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG).

In some embodiments, the binding agent formulation is selected from the following structure:

, And .

On the other hand, the present patent application provides a binding agent preparation that contains the binding agent described in this application.

In some embodiments, the binding agent formulation is selected from the following structure: Z'-A-S*-RL-D and Z'-A-S*-RL-Y-D; where Z' indicates a subclone precursor, and the letter A indicates a linkage or shared unit. S* indicates a linkage or separating agent, and Y indicates a spacer subunit; and the letter D indicates a drug unit.

In some embodiments, where the subclone precursor Z' is selected from (maleimide-N-yl)-, (pyrrolidine-2,5-diketone-N-yl)-, M-CH ₂ -C(=O)-NH-, and HS-(CH ₂ ) _1-5 -C(=O)-, where M indicates halogen.

In some embodiments, where the dosage unit includes a cytotoxic agent.

In some embodiments, where the dosage unit comprises: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicines, colcimides, combretastatins, cryptophycins, discodemolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodiazepines (PBD), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vinca alkaloids or derivatives thereof.

In some embodiments, where the dosage unit comprises a DNA topoisomerase I inhibitor.

In some embodiments, the dosage unit comprises camptothecin or a derivative thereof.

In some embodiments, where the medicinal unit combines the following structure:

Dxd

The nitrogen atom of the amino group at position 1 is the group in the position of attachment.

In some embodiments, where the binding agent preparation comprises the following structure:

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd; And

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

Where,

(Maleimide-N-yl)- has the following structure: , where the nitrogen atom represents a group in the attachment position;

(Pyrrolidine-2,5-diketone-N-yl)- has the following structure: , where the nitrogen atom represents a group in the attachment position;

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG) or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG);

Dxd contains the following structure: , where the nitrogen atom of the amino group in position 1 represents a group in the position of attachment;

M indicates a halogen atom.

In some embodiments, where the binding agent preparation is selected from the following structure:

; And .

On the other hand, the present patent application proposes a conjugate that includes a binding agent in the present application.

In some embodiments, the conjugate comprises a ligand.

In some embodiments, the ligand targets a cell surface receptor or tumor-associated antigen.

In some embodiments, the described cell surface receptor or tumor-associated antigen may include HER2, HER3, claudin18.2, folate receptor α (FRα), BCMA, PSMA, TROP-2, CD19, CD20, CD22, CD30, CD79b, EGFR, c-Met, or CEACAM5.

In some embodiments, the ligand comprises an antibody or an antigen-binding fragment thereof.

In some embodiments, the antibody comprises a monoclonal antibody, a polyclonal antibody, a dimer, a polymer, a multispecific antibody, an intact antibody, an antibody fragment, a human antibody, a humanized antibody, or a chimeric antibody.

In some embodiments, the antigen-binding fragment comprises a Fab, Fab', Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.

In some embodiments, where the antibody is monoclonal.

In some embodiments, the antibody comprises a human antibody, a humanized antibody, or a chimeric antibody.

In some embodiments, the conjugate comprises a ligand-drug conjugate and has the following structure: C-(LD) _m , C denotes a ligand unit, L denotes a binding agent, D denotes a drug unit, and m denotes any number from 1 to 10.

In some embodiments, the ligand-drug conjugate is an antibody-drug conjugate (ADC) and has the following structure: Ab-(LD) _m , where Ab is an antibody or antigen-binding fragment thereof, L is a binding agent, D is a unit of drug, and m is any number from 1 to 10.

In some embodiments, where the dosage unit includes a cytotoxic agent.

In some embodiments, where the dosage unit comprises: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicines, colcimides, combretastatins, cryptophycins, discodemolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodiazepines (PBD), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vinca alkaloids or derivatives thereof.

In some embodiments, where the dosage unit comprises a DNA topoisomerase I inhibitor.

In some embodiments, the dosage unit comprises camptothecin or a derivative thereof.

In some embodiments, where the medicinal unit combines the following structure:

Dxd

The nitrogen atom of the amino group at position 1 is the group in the position of attachment.

In some embodiments, wherein the antibody-drug conjugate is selected from the following structure:

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ; And

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Where,

(succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG) or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG);

Dxd contains the following structure: , where the nitrogen atom of the amino group in position 1 represents a group in the position of attachment;

Ab denotes an antibody or its antigen-binding fragment, and m denotes any number from 1 to 10.

In some embodiments, wherein the antibody-drug conjugate is selected from the following structure:

; And ; where Ab denotes an antibody or its antigen-binding fragment, and m denotes any number from 1 to 10.

In some embodiments, the antibody or antigen-binding fragment thereof comprises: an anti-HER2 antibody, an anti-HER3 antibody, an anti-claudin18.2 antibody, an anti-FRα antibody, or fragments that bind its antigens.

On the other hand, the present patent application proposes a combination of drugs that comprises a conjugate described in this patent application and an additional carrier that is pharmaceutically acceptable.

On the other hand, the present patent application provides for the use of a conjugate or combination of drugs as described in this patent application in the preparation of a medicament for the treatment of diseases or disease states.

In some embodiments, where the diseases or conditions include tumors.

In some embodiments, wherein the diseases or conditions include solid tumors and hematological tumors.

In some embodiments, wherein the solid tumor comprises: gastric cancer, gastric cancer with metastases to the peritoneum, liver cancer, leukemia, kidney cancer, tumor, lung cancer, small bowel cancer, bone cancer, prostate cancer, colorectal cancer, breast cancer, colon cancer, cervical cancer, ovarian cancer, lymphatic cancer, nasopharyngeal cancer, adrenal tumor, bladder tumor, non-small cell lung cancer (NSCLC), glioma, endometrial cancer, or a combination thereof.

In some embodiments, wherein the hematological tumors include: acute myeloid leukemia (AML), multiple myeloma (MM), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, or a combination thereof.

On the other hand, the present patent application provides a method for treating diseases or disease states in patients in need thereof, comprising administering to the patients an effective dose of the conjugate as described in this application.

In some embodiments, where the diseases or conditions include tumors.

In some embodiments, wherein the diseases or conditions include solid tumors and hematological tumors.

In some embodiments, wherein the solid tumor comprises: gastric cancer, gastric cancer with metastases to the peritoneum, liver cancer, leukemia, kidney cancer, tumor, lung cancer, small bowel cancer, bone cancer, prostate cancer, colorectal cancer, breast cancer, colon cancer, cervical cancer, ovarian cancer, lymphatic cancer, nasopharyngeal cancer, adrenal tumor, bladder tumor, non-small cell lung cancer (NSCLC), glioma, endometrial cancer, or a combination thereof.

In some embodiments, wherein the hematological tumors include: acute myeloid leukemia (AML), multiple myeloma (MM), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, or a combination thereof.

Those skilled in the art can readily appreciate other aspects and advantages of the present invention from the detailed description set forth below. The detailed description below depicts and describes only an exemplary embodiment of this patent application. Those skilled in the art will appreciate that the contents of this patent application permit them to modify the disclosed specific embodiments without departing from the spirit and scope of the invention described in this patent application. Accordingly, the descriptions in the drawings and instructions accompanying this application are illustrative only and are not limiting.

From description to drawings

Specific features of the invention described in this patent application are set forth in the appended claims. The features and advantages of the invention described in this application can be better understood by using the approximate embodiments and drawings described below. A brief description of the drawings is as follows:

Figure 1A shows a HIC-HPLC chromatogram of HER3-ADC04 as described in embodiment 1 of this patent application.

Figure 1B shows a HIC-HPLC chromatogram of HER3-ADC07 as described in Embodiment 1 of this application.

Figure 1C shows a HIC-HPLC chromatogram of HER3-ADC08 as described in embodiment 1 of this application.

Figure 1D shows a HIC-HPLC chromatogram of HER3-ADC09 as described in Embodiment 1 of this application.

Figure 1E shows the HIC-HPLC chromatogram of Claudin18.2-ADC08 as described in Embodiment 1 of this application.

Figure 1F shows the HIC-HPLC chromatogram of Claudin18.2-ADC09 as described in Embodiment 1 of this application.

Figure 1G shows a HIC-HPLC chromatogram of FRα-ADC08 as described in Embodiment 1 of this application.

Figure 1H shows a HIC-HPLC chromatogram of FRα-ADC09 as described in Embodiment 1 of this application.

Figure 2A shows the in vitro killing effect of HER3-ADC04 on SW620 human colon cancer cells, as described in this patent application.

Figure 2B shows the in vitro killing activity of all -Dxd binding agent formulations on SW620 human colon cancer cells as described in this patent application.

Figure 3 shows the in vitro killing activity of anti-HER3 ADCs containing various binding agents on SW620 human colon cancer cells, as described in this patent application.

Figure 4 shows the in vitro killing activity of anti-HER3 ADCs containing various binding agents on CHO-K1 colon cancer target cells, as described in this patent application.

Figure 5 shows the in vitro killing activity of anti-HER3 ADC on SW620 target cells after incubation in plasma, as described in this patent application.

Figure 6 shows the effect of anti-HER3 ADC treatment on the SW620 transplanted tumor model in this patent application.

Figure 7 shows the changes in mouse weight during treatment with anti-HER3 ADC in the SW620 transplanted tumor model, as described in this patent application.

Figure 8 shows the in vitro killing activity of anti-Claudin18.2 ADC on human gastric cancer cells NUGC4, as described in this patent application.

Figure 9 shows the in vitro killing activity of an anti-Claudin18.2 ADC on human gastric cancer cells NUGC4 recombinant overexpressing the Claudin18.2 molecule, as described in this patent application.

Figure 10 shows the in vitro killing activity of an anti-Claudin18.2 ADC on HEK293 cells recombinantly overexpressing the Claudin18.2 molecule, as described in this patent application.

Figure 11 shows the in vitro killing activity of anti-Claudin18.2 ADC on HEK293 overexpressing the Claudin18.2 molecule after incubation in plasma, as described in this patent application.

Figure 12 shows the growth inhibitory effect of anti-claudin18.2 ADC in the NUGC4 transplant gastric cancer model as described in this patent application.

Figure 13 shows the in vitro killing activity of anti-FRα ADC on OVCAR3 human ovarian cancer cells, as described in this patent application.

Figure 14 shows the in vitro killing effect of anti-FRα ADC on anti-FRα ADC after incubation in plasma, as described in this patent application.

Figure 15 shows the growth inhibitory effect of anti-FRα ADC in the subcutaneously transplanted tumor model in OVCAR3 mice, as described in this patent application.

Design

Specific embodiments of the invention according to this patent application are described as follows. Those familiar with this technology will readily understand other advantages and effects of this application from the contents disclosed in the instructions.

Terms and definitions

In this patent application, the terms "antibody-drug conjugate (ADC)", "antibody-drug conjugate (ADC)", and "ADC" are used interchangeably. They generally refer to one or more therapeutic compounds (e.g., Dxd) that are linked to one or more antibodies or antigen-binding fragments and are defined by the following general formula: Ab-(LD) _m , where Ab=antibody or antigen-binding fragment, L=binding agent, D=drug unit, and m=the number of drug parts per antibody or antigen-binding fragment, which may be any number from 1 to 10. In some embodiments, the binding agent L may include a lysable moiety between the antibody or antigen-binding fragment and the drug unit. A typical lysable compound is described and demonstrated in the article.

In this patent application, the term "antibody" is generally used in its broadest sense and specifically encompasses a monoclonal antibody, a polyclonal antibody, a dimer, a polymer, a multispecific antibody (e.g., a bispecific antibody), and an antibody fragment, as long as they exhibit the desired biological activity (Millere et al. (2003) Jour. of Immunology 170:4854-4861). The antibody may be a murine antibody, a human antibody, a humanized antibody, a chimeric antibody, or an antibody derived from another species. Non-restrictive "antibodies" may typically include proteins with at least two heavy chains (HC) and two light chains (LC) linked to each other through disulfide bonds or their antigen-binding fragments. Each heavy chain contains a heavy chain variable region (VH) and a heavy chain constant region. In some naturally occurring IgG, IgD, and IgA antibodies, the heavy chain constant region comprises three domains (CH1, CH2, and CH3). In some naturally occurring antibodies, each light chain contains a variable light chain region (VL) and a constant light chain region. The constant light chain region comprises a domain (CL). The VH and VL regions can be further divided into hypervariable regions known as complementary determining regions (CDRs), which alternate with more conserved regions called framework regions (FRs). Each VH and VL region contains three CDRs and four framework regions (FRs), arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Each of the variable domains of natural heavy and light chains consists of four FR regions (HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4), most of which utilize a β-sheet configuration and are connected by three CDRs to form a ring junction, and in some cases form part of a β-sheet structure. The CDRs in each chain are tightly packed across the FR region and form antigen-binding sites with CDRs from another chain. The constant region of an antibody can mediate the binding of immunoglobulins to tissues or host factors, including various immune system cells (e.g., nerve terminal cells) and the first component (Clq) of the classical complement system.

In this patent application, the terms "antigen-binding fragment" or "functional fragment of an antibody" (also referred to in the article as a "targeting portion" or "antigen-binding portion") generally refer to the portion of an antibody molecule that contains the amino acid responsible for the specific binding between the antibody and an antigen. The portion of an antigen that is recognized and specifically bound by an antibody is called an "epitope", as described above. An antigen-binding fragment may typically comprise a variable region of the light chain (VL) and a variable region of the heavy chain (VH) of the antibody; however, it does not necessarily include both. Embodiments of an antigen-binding fragment of an antibody include the following: (1) Fab fragments and monovalent fragments with VL, VH, constant light chain (CL), and CH1 domains; (2) F(ab') ₂ fragments and divalent fragments of two Fab fragments linked by a disulfide bridge at the hinge region; (3) Fd fragments with two VH and CH1 domains; (4) Fv fragments with the VL and VH domains of a single-arm antibody, (5) dAb fragments (Ward et al., “Binding Activities of a Repertoire of Single Immunoglobulin Variable Domains Secreted From Escherichia coli,” Nature 341:544-546 (1989), incorporated by reference into this application) with its VH domain; (6) isolated complementary determining region (CDR); (7) single-chain Fv (scFv), e.g., obtained from an scFV library. Although the two structural domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be joined recombinantly using synthetic linkers. Binding agents are synthesized in a manner that allows them to be produced as a single protein chain in which the VL and VH regions are paired to form a monovalent molecule (called single-chain Fv (scFv)) (Huston et al., “Protein Engineering of Antibody Binding Sites: Recovery of Specific Activity in an Anti-Digoxin Single-Chain Fv Analogue Produced in Escherichia coli,” Proc. Natl. Acad. Sci. USA 85:5879–5883 (1988)); and (8) VHH. "VHH" includes variable antigen-binding structural domains of heavy chain antibodies from the camelid family (camelids, dromedaries, jaguars, alpacas, etc.) (Nguyen VK et al., 2000, The EMBO Journal, 19, 921-930; Muyldermans S., 2001, J Biotechnol., 74, 277-302 and Vanlandschoot P. et al., 2011, Antiviral Research 92, 389-407). These antibody fragments are obtained by routine methods known to those skilled in the art. The function of these fragments can be assessed in the same way as that of intact antibodies.

In this patent application, the term "variable" generally refers to the fact that some portion of the variable structural domain sequence of an antibody varies so significantly that it shapes the binding and specificity of different antibodies to their specific antigens. However, the variability is not distributed uniformly across the variable region of the antibody. It is concentrated in three segments of the variable regions of the light and heavy chains, known as the complementary decision region (CDR) or the highly variable region (HVR). The more conserved portion of the variable domain is called the frame (FR). In this field, antibody CDRs can be defined in various ways, such as the Kabat definition rule based on sequence variability (Kabat et al., Protein Sequence in Immunology, Vol. V, National Institutes of Health, Bethesda, MD (1991)), the Chothia definition rule based on the location of structural loop regions (A1-Lazikani et al., JMol Biol 273:927-48, 1997), and the KABAT definition rule based on the IMGT ontology concept (IMGT-ONTOLOGY). and the IMGT scientific map rules.

In this patent application, the term "monoclonal antibody" generally refers to an antibody obtained from a cluster of substantially homogeneous antibodies, meaning that the individual antibodies in the cluster are identical except for a small number of possible naturally occurring mutations. Monoclonal antibodies are typically highly specific for a single antigenic site. Moreover, unlike conventional polyclonal antibody preparations, which typically contain different antibodies directed against different determinants, each monoclonal antibody is directed against a single determinant of the antigen. In addition to their specificity, monoclonal antibodies have the following advantage: they can be synthesized in hybrid culture and are not contaminated with other immunoglobulins. The modifier "monoclonal" characterizes antibodies obtained from a substantially homogeneous population of antibodies. This is not construed as requiring production of the antibodies by any particular method. For example, the monoclonal antibodies used in this patent application can be produced in hybridoma cells or by recombinant DNA techniques.

In this patent application, the term "chimeric antibody" generally refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species. Generally, the variable region is derived from antibodies from experimental animals, such as rodents ("parental antibodies"), and the constant region is derived from human antibodies. This makes the resulting chimeric antibodies less likely to elicit an adverse immune response in humans compared to parental antibodies (e.g., those of mouse origin).

In this patent application, the term "humanized antibody" generally refers to an antibody in which some or all amino acids outside the CDR region of a non-human antibody (such as a murine antibody) are replaced with corresponding amino acids derived from human immunoglobulins. Minor additions, deletions, insertions, substitutions, or modifications of amino acids within the CDR region may also be permissible as long as they maintain the antibody's ability to bind a specific antigen. A humanized antibody may further comprise at least a portion of a constant region of a human immunoglobulin. "Humanized antibodies" retain antigen specificity similar to that of the parent antibodies. "Humanized" forms of non-human antibodies (such as murine antibodies) may, to a minimal extent, contain chimeric antibodies derived from non-human immunoglobulin sequences. In some cases, human immunoglobulin residues (the recipient antibody) in the CDR region may be replaced with residues in the CDR region from a non-human species (the donor antibody) (e.g., mouse, rat, rabbit, or non-human primate) that have the desired properties, similarities, and/or capabilities. In some cases, human immunoglobulin residues in the FR region may be replaced with corresponding non-human residues. Furthermore, a humanized antibody may contain amino acid modifications that are not present in the recipient or donor antibody. These modifications may be made to further improve antibody properties, such as binding affinity.

In this patent application, the term "fully human antibody" or "human antibody" generally refers to an antibody expressed in a genetically engineered antibody deficient in the antibody gene by transferring the human gene encoding the antibody into an animal. All parts of the antibody (including the variable and constant regions of the antibody) are encoded by human genes. Fully human antibodies can significantly reduce the immune side effects caused by heterologous antibodies in the human body. Methods for producing fully human antibodies in this area may include phage display technology, transgenic mouse technology, ribosome display technology, RNA-polypeptide technology, etc.

In this patent application, when referring to the interaction of a binding molecule (e.g., an antibody) with its binding partner (e.g., an antigen), the term "specific binding" generally means that the interaction is dependent on the presence of a specific structure (e.g., an antigenic determinant or epitope) on the binding partner. In other words, the antibody will preferentially bind to or recognize the binding partner even if the binding partner is present in a mixture of other molecules or organisms. The establishment of binding may be mediated by covalent or non-covalent interactions, or a combination of both. In other words, the term "specific binding" generally refers to immunospecific binding to antigenic determinants or epitopes without immunospecific binding to other antigenic determinants or epitopes. The binding molecule of an immunospecific binding antigen can bind other peptides or polypeptides with lower affinity, as determined by radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), BIACORE, or other assays known in the art. Binding molecules of immunospecific binding antigens or their fragments may react well with related antigens with the same epitope. In some cases, binding molecules of immunospecific binding antigens or their fragments do not cross-react with other antigens.

In this patent application, the term "binding agent," "compound," "binding agent," or "L" generally refers to any chemical moiety that covalently provides a linkage (usually a drug) to another moiety (such as an antibody or antigen-binding fragment). The compound may be sensitive to or substantially resistant to acid-induced lysis, peptidase-induced lysis, light-induced lysis, esterase-induced lysis, and/or disulfide bond lysis under conditions that preserve the activity of the compound or antibody.

In this patent application, the term "drug unit" or "D" generally refers to any compound with the desired biological activity and functional group reactivity that can be used to incorporate a drug into the conjugate of this application. In some embodiments, the drug unit is a cytotoxic drug for use in cancer therapy. The compound contains proteins or polypeptides with the desired biological activity, such as toxins (abrin, ricin A, pseudomonas exotoxin, diphtheria, etc.) and other relevant proteins, including tumor necrosis factor, α-interferon, β-interferon, nerve growth factor, platelet-derived growth factor, tissue plasminogen activator and biological response modifiers such as lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) or other growth factors.

In this patent application, the term "cytotoxic agent" generally refers to a substance that inhibits cell function and/or causes cell death or destruction. A cytotoxic agent includes, but is not limited to, radioisotopes (such as At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and Lu), chemotherapeutic agents or drugs (such as methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine and etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other inserts). It also includes growth inhibitors, enzymes and fragments thereof (such as nucleotide-splitting enzymes), antibiotics, toxins (such as small molecule toxins or enzymatically active toxins derived from bacteria, fungi, plants or animals), including fragments and/or variants thereof, and various antitumor or anticancer agents, which are described below.

In this patent application, the term "pharmaceutically acceptable form" generally refers to a form of the disclosed compound, including, but not limited to, its pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, and isotopically labeled derivatives. In one embodiment, "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, prodrugs, and isotopically labeled derivatives. In some embodiments, "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, prodrugs, and isotopically labeled derivatives.

The binding agents, drug binding agents, and conjugates described in this patent application may comprise any of their pharmaceutically acceptable forms, such as pharmaceutically acceptable salts, esters, hydrates, solvates, polymorphs, isomers, prodrugs, or isotopically labeled derivatives. The aforementioned pharmaceutically acceptable forms are also included within the scope of this patent application.

In this patent application, the term "pharmaceutically acceptable salt" generally refers to a pharmaceutically acceptable organic or inorganic salt of a compound (such as a drug, binding agent, or conjugate). In some aspects, the compound may contain at least an amino group and thus can form acid addition salts with the amino group. Examples of salts include, but are not limited to, sulfates, trifluoroacetates, citrates, acetates, oxalates, chlorides, bromides, iodides, nitrates, bisulfates, phosphates, hydrogen phosphates, isonicotinic acid esters, lactates, salicylates, citrates, tartrates, oleates, tannates, pantothenates, bitartrates, ascorbates, succinates, maleates, gentates, fumarates, gluconates, glucuronates, saccharates, formates, benzoates, glutamates, mesylates, esylates, benzenesulfonates, P-toluenesulfonates, and pamoates (1,1'-methylenebis-(2-hydroxyl-3-naphthoate)). Pharmaceutically acceptable salts may include the introduction of another molecule such as an acetate ion, a succinate ion, or other counterions. A counterion can be any organic or inorganic moiety that stabilizes the charge of the parent compound. Furthermore, pharmaceutically acceptable salts may contain more than one charged atom. Embodiments in which multiple charged atoms are part of pharmaceutically acceptable salts may also contain multiple counterions. Thus, pharmaceutically acceptable salts may contain one or more charged atoms and/or one or more counterions.

In this patent application, the term "derivative" generally refers to a chemical compound or molecule obtained from a parent compound through one or more chemical reactions.

In this patent application, the term "prodrug" generally refers to a compound with little or no biological activity. This compound is then transformed into a compound with higher biological activity in the laboratory through a chemical or biological process (e.g., a chemical reaction or enzymatic biotransformation).

In this patent application, the term "isotopically labeled compound" or "isotopically labeled derivative" generally refers to the presently described compounds in which one or more atoms are replaced by an atom with an atomic mass or mass number different from that typically found in nature, including the corresponding pharmaceutical preparations described herein: salts and prodrugs. Isotopic variants that can be introduced into the presently described compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, and ³⁶ Cl, respectively.

In this patent application, the term "pharmaceutically acceptable carrier" generally refers to one or more non-toxic substances that do not interfere with the biological efficacy of the active ingredient. Formulations of this type may typically contain a salt, a buffering agent, a preservative, a compatible carrier, an adjuvant, and optionally other therapeutic agents. This type of pharmaceutically acceptable formulation may also typically contain a compatible solid or liquid filler, diluent, or capsule coating material suitable for human administration. Non-limiting pharmaceutically acceptable carriers may include liquids such as water, saline, glycerol, and ethanol. These carriers may also contain excipients such as a wetting or emulsifying agent, a pH buffer, etc.

In this patent application, the term "alkyl" or "alkyl group" generally refers to a fully saturated straight, branched, or cyclic hydrocarbon chain. In some embodiments, alkyl may contain 1-8 carbon atoms ("C ₁ -C ₈ alkyl"). In some embodiments, alkyl may contain 1-6 carbon atoms ("C ₁ -C ₆ alkyl"). In some embodiments, alkyl contains 1-3 carbon atoms. In other embodiments, alkyl contains 2-3 carbon atoms, while in other embodiments, alkyl contains 1-2 carbon atoms.

In this patent application, the term "alkylalkoxy" refers to alkyl substituted with alkoxy.

In this patent application, the term "alkoxy" refers to an alkyl, as previously defined, attached to the main carbon chain through an oxygen atom ("alkoxy").

In this patent application, the term "alkylalkoxy" refers to alkyl substituted with alkoxy.

In this patent application, the term "hydroxy" or "hydroxyl" refers to -OH.

In this patent application, the terms "carbocyclyl" or "carbocyclyl" include both aromatic groups (such as aryl) and non-aromatic groups (such as cycloalkyl). In some embodiments, the carbocyclic group contains 3-10 carbon atoms ("3- to 10-membered carbocyclyls"). In some embodiments, the carbocyclic group contains 3-8 carbon atoms ("3- to 8-membered carbocyclyls"). In some embodiments, the carbocyclic group contains 3-6 carbon atoms ("3- to 6-membered carbocyclyls"). In some embodiments, the carbocyclic group contains 3-5 carbon atoms ("3- to 5-membered carbocyclyls").

In this patent application, the term "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms.

In this patent application, the term "halogen" refers to any halogen group such as -F, -Cl, -Br, or -I.

In this patent application, the terms "heterocycle or heterocyclyl or heterocyclic" and "heterocyclyl" refer to a single-ring heterocycle, a bicyclic heterocycle, or a tricyclic heterocycle containing at least one heteroatom in the ring. The terms "heterocycle or heterocyclyl or heterocycle" and "heterocyclyl" include heteroaryl. "Heteroaryl" refers to a cyclic moiety with one or more closed rings and one or more heteroatoms (oxygen, nitrogen, or sulfur) in at least one of the rings, wherein at least one of the rings is an aromatic ring, and the ring or ring type may be independently fused and/or bridged. Embodiments include, but are not limited to, phenyl, thienyl, triazolyl, pyridyl, pyrimidinyl, pyridazinyl, and pyrazinyl.

In this patent application, the term "PEG unit" generally refers to an organic moiety composed of repeating ethyleneoxy subunits (PEG or PEG subunits) and may be polydisperse, monodisperse, or discrete (equivalent to ethyleneoxy subunits of a discrete number). Polydisperse PEG is a heterogeneous mixture in size and molecular weight, while monodisperse PEG is typically purified from a heterogeneous mixture and therefore has a uniform chain length and molecular weight. A PEG unit in this patent application may comprise one or more polyethyleneglycol chains, and each polyethyleneglycol chain is composed of one or more ethyleneoxy subunits covalently in adhesive association with one another. For example, the polyethyleneglycol chains may be linked together in a linear, branched, or star-shaped configuration.

In this patent application, the term "substituted," whether preceded by the term "additional" or not, generally indicates that one or more hydrogen atoms of the designated moiety have been replaced by the corresponding substituents. Unless otherwise specified, an "additionally substituted" group may contain the corresponding substituents at each substitutable position of the group, and when more than one position in any given structure can be substituted by more than one substituent selected from the specified group, all substituents at a position may be either the same or different. According to the present invention, it is contemplated that a combination of substituents will result in the formation of stable or chemically feasible compounds.

Those skilled in the art will understand that the terms "substitution," "substituted," or "nonexistent" include the following implicit limiting conditions: whether or not substitution is based on the permissible valence of the substituted atom and the substituent, and whether or not substitution is nonexistent forms a stable compound, for example, a stable compound undergoes transformation non-spontaneously through rearrangement, cyclization, elimination, etc. For the purposes of the present invention, nitrogen heteroatoms may contain hydrogen substituents and/or any permissible substituent of organic compounds described in the article that satisfies the valence number of the heteroatoms.

In this patent application, the term "prevention and/or treatment" not only includes preventing and/or treating a disease, but generally includes preventing the onset of a disease, slowing or reversing the progression of a disease, preventing or slowing the progression of a disease, an attack of one or more symptoms associated with a disease, reducing and/or alleviating one or more symptoms associated with a disease, reducing the severity and/or duration of a disease and/or any symptoms associated with a disease, and/or preventing a disease and/or preventing a further increase in the severity of any symptoms associated with a disease, preventing, reducing or reversing any physiological damage caused by a disease, as well as any pharmacological effects that are generally beneficial to the patient undergoing treatment. The combination of this patent application forms a possible therapeutic agent without achieving a complete cure or elimination of any symptoms or manifestations of the disease. As recognized in the relevant art, a drug used as a therapeutic agent can reduce the severity of a given disease state, but does not necessarily eliminate all manifestations of the disease to be considered a useful therapeutic agent. Similarly, prophylactic treatment is a genuine preventative measure that does not necessarily have to be completely effective in preventing the onset of a disease. It aims to reduce the impact of the disease on patients, for example, by reducing the number or severity of symptoms, increasing the effectiveness of another treatment, or achieving another beneficial effect, or by reducing the likelihood of the disease occurring or worsening.

In this patent application, the term "administration" generally refers to the delivery of proteins (including immunoglobulins) to patients or animals in need by any route known in the art. The drug carrier and the drug itself or its composition are also well known in the art. Routes of administration may include intravenous, intramuscular, intradermal, subcutaneous, transdermal, mucosal, intratumoral, or mucosal.

In this patent application, the term "contact" generally refers to two or more different types of substances brought into contact in any order, by any method, and for any duration. When applied to cells, the term "contact" generally refers to the method of delivering an antibody or antigen-binding fragment thereof, a drug, a drug binding agent, a conjugate, and/or a combination of drugs of this application to or in close proximity to a target cell, where delivery may occur in vivo, extracorporeally, or may involve the use of a recombinant carrier system. "Contact" may occur in vivo, extracorporeally, or in vitro.

In this patent application, the term "effective amount" or "effective dose" generally refers to an amount sufficient to achieve, or at least partially achieve, the desired effect. An "effective therapeutic amount" or "effective therapeutic dose" of a drug or therapeutic agent generally refers to any amount of the drug that, when used alone or in combination with another therapeutic agent, promotes cure of a disease (as evidenced by a reduction in the severity of the disease), its symptoms, an increase in the incidence and duration of symptom-free periods, or the prevention of damage or disability caused by the disease. The terms "effective prophylactic amount" or "effective prophylactic dose" of a drug generally refers to an amount of the drug that inhibits the development or recurrence of a disease when administered alone or in combination with another therapeutic agent to patients at risk of developing or recurring the disease. A variety of methods known to those skilled in the art can be used to evaluate the ability of therapeutic or prophylactic agents to promote disease regression or inhibit the progression or recurrence of disease, for example, in humans undergoing clinical trials and in animal model systems, by predicting efficacy in humans, or by measuring the activity of agents in in vitro assays.

In this patent application, the term "tumor" generally refers to the growth and spread of all neoplastic cells, including malignant and benign cells, as well as all precancerous and cancerous cells and tissues. The terms "cancer," "carcinoma," "cell proliferative symptoms," "proliferative symptoms," and "tumor" are not mutually exclusive when used in this article. In this patent application, a tumor may be a solid tumor or a non-solid tumor.

In this patent application, the term "patient" generally refers to humans or animals (including mammals) requiring diagnosis, prognosis, amelioration, prevention, and/or treatment of diseases, such as humans, non-human primates (monkeys, gibbons, gorillas, chimpanzees, orangutans, and macaques), companion animals (dogs and cats), farm animals (poultry such as chickens and ducks, horses, cows, goats, sheep, and pigs), and experimental animals (mice, rats, rabbits), as well as guinea pigs. Human patients include fetuses, neonates, infants, adolescents, and adult patients. Patients may include animal disease models.

In this patent application, the terms "including," "comprising," "having," "may," "comprising," and variations thereof are generally intended to be transitional phrases, terms, or open-ended words and do not exclude the possibility of additional actions or structures. The term "consisting of" generally means that there may be no further components (or, similarly, functions, integers, steps, etc.). Unless otherwise expressly stated in the text, singular forms such as "a," "an," and "the" in English, and the words "one," "one type," and "specified/defined" in Russian, generally include the plural form of the word in question.

In this patent application, the term "approximately" generally refers to approximately, in the region of, approximately, or around. When the term "approximately" is used to indicate a range of numerical values, a limiting or specific value is indicated from which the claimed value may differ by up to 10%. Therefore, the term "approximately" may be used to indicate deviations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the specified value.

Resume

Binding agent

This patent application provides a novel conjugating coupling agent structure that comprises an amino acid unit (RL), wherein RL comprises: -Val-Ala-Q-Gly-Phe-Gly-, wherein Q indicates a linkage, a single amino acid, or a dipeptide.

In some embodiments, RL may be lysed by enzymes.

In some embodiments, where RL may be lysed by cathepsin B.

In some embodiments, where Q indicates a connection.

In some embodiments, where Q indicates -Ser- or -Ser-Ser-.

In some embodiments, where RL combines -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG), or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG).

In some embodiments, the binding agent is selected from the following sequences: -Z-A-S*-RL- and -Z-A-S*-RL-Y-; where Z denotes a subclone and A denotes a linkage or co-unit, where S* denotes a linkage or separating agent, and Y denotes a spacer subunit.

In some embodiments, wherein Z comprises the following structure: -(succinimide-3-yl-N)-, -CH ₂ -C(=O)-NH-, or -C (=O)-;

where (succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

In some embodiments, where A is selected from the following structure:

-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ heteroalkylene-, -C ₃ -C ₈ carbocycloalkyl-, -O-(C ₁ -C ₈ alkylene)-, -arylene-, -C ₁ -C ₁₀ alkylene-arylene-, -arylene-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-, -C ₃ -C ₈ heterocyclyl-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-, -C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ heteroalkylene-C(=O)-, -C ₃ -C ₈ carbocycloalkyl-C(=O)-, -O-(C ₁ -C ₈ alkylene)-C(=O)-, -arylene-C(=O)-, -C ₁ -C ₁₀ alkylene-arylene-C(=O)-, -arylene-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-C(=O)-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₃ -C ₈ heterocyclyl-C(=O)-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-C(=O)-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-C(=O)-, -C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ heteroalkylene-NH-, -C ₃ -C ₈ carbocycloalkyl-NH-, -O-(C ₁ -C ₈ alkylene)-NH-, -arylene-NH-, -C ₁ -C ₁₀ alkylene-arylene-NH-, -arylene-C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-NH-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-NH-, -C ₃ -C ₈ heterocyclyl-NH-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-NH-, -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-NH-, -C ₁ -C ₁₀ alkylene-S-, -C ₁ -C ₁₀ heteroalkylene-S-, -C ₃ -C ₈ carbocycloalkyl-S-, -O-(C ₁ -C ₈ alkylene)-S-, -arylene-S-, -C ₁ -C ₁₀ alkylene-arylene-S-, -arylene-C ₁ -C ₁₀ alkylene-S-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ carbocycloalkyl)-S-, -(C ₃ -C ₈ carbocycloalkyl)-C ₁ -C ₁₀ alkylene-S-, -C ₃ -C ₈ heterocyclyl -S-, -C ₁ -C ₁₀ alkylene-(C ₃ -C ₈ heterocyclyl)-S- or -(C ₃ -C ₈ heterocyclyl)-C ₁ -C ₁₀ alkylene-S-;

wherein A is further replaced by a basic unit (BU) that contains -( _CH2 ) _xNH2 , _- ( _CH2 ) _xNHRa ^, or -( _CH2 ) _xNRa2 where x is any integer from 1 to ₄ ; further, each R ^a is independently selected from C1 _{- C6} alkyl and C1 _{- C6} haloalkyl, or two R ^a groups are combined ^with nitrogen attached thereto to form a 4-6-membered heteroaryl or azetidinyl, pyrrolidyl, or piperidyl group.

In some embodiments, A is selected from the following structure: -(C ₁ -C ₅ ) alkylene-C(=O)-, wherein the alkylene portion of A is further replaced by a basic unit (BU).

For example, Z may contain -(succinimide-3-yl-N)-, while A may be -(C ₁ -C ₁₀ ) alkylene-C(=O)-, and ZA may contain the following structure:

, where a specifies an integer from 1 to 8. For example, where a can be an integer from 2 to 5.

In another example, Z-A- might contain the following structure:

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ -C(=O)-

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

-CH ₂ C(=O)NH-CH ₂ -C(=O)-

- CH ₂ C(=O)NH-CH ₂ CH ₂ -C(=O)-

- CH ₂ C(=O)NH-CH ₂ CH ₂ CH ₂ -C(=O)-

- CH ₂ C(=O)NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

- CH ₂ C(=O)NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

-C(=O)-cyc.Hex(1,4)-CH ₂ -(N-ly-3-diminiccuS)-

-C(=O)-Aryl-CH ₂ -(N-ly-3-diminiccuS)-

-C(=O)-cyc.Het-CH ₂ -(N-ly-3-diminiccuS)-

- C(=O)-CH ₂ CH ₂ -C(=O)-

- C(=O)-CH ₂ CH ₂ CH ₂ -C(=O)-

-C(=O)-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

-C(=O)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-

-C(=O)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-, etc.

(Aryl and cyclyl refer to a divalent aromatic hydrocarbon group. cyc.Het refers to a divalent cyclic heterocyclyl.)

In some embodiments, where Y comprises the structure shown in -NH-(CH ₂ )n ¹ -La-Lb-Lc-, where La is -C(=O)-NH-, -NR ¹ -(CH ₂ )n ² -, -O-, or a single bond, and R ¹ is H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkylene carboxyl, or C ₁ -C ₆ alkylene hydroxyl.

Lb indicates ^-CR2 ( ^-R3 )- or one bond, where ^R2 and ^R3 independently indicate H, _-NH2 , _C1 - _C6 alkylene, _C1 - _C6 alkylene-amino, _C1 - _C6 alkylene- ^carboxyl , or C1- _C6 alkylene-hydroxyl, and _R2 and ^R3 are not simultaneously _-NH2 .

n ¹ indicates an integer from 0 to 6;

Lc indicates -CH ₂ - or -C(=O)-.

In some embodiments, where Y comprises NH-( _CH2 ) ⁿ¹ -O-( _CH2 ) ⁿ² -C(=O)-, where ⁿ¹ or ⁿ² is independently any integer from 0 to 5. Spacers may be functional groups that facilitate the attachment of RL to the drug units or may provide additional structural components to further facilitate the release of drug units from the rest of the conjugate (e.g., self-removal of para-aminobenzyl (PAB) components).

In some embodiments, where Y combines -NH-CH ₂ -C(=O)-, -NH-CH ₂ CH ₂ -C(=O)-, -NH-(CH ₂ ) ₃ -C(=O)-, -NH-(CH ₂ ) ₄ -C(=O)-, -NH-(CH ₂ ) ₅ -C(=O)-, -NH-CH ₂ -O-CH ₂ -C(=O)-, or -NH-(CH ₂ ) ₂ -O-CH ₂ -C(=O)-.

For example, Y might contain the following structure:

-NH-CH ₂ -

-NH-CH(-Me)-

-NH-C(-Me) ₂ -

-NH-CH ₂ -CHMe-

-NH-CH(-CH ₂ OH)-

-NH-CH(-CH ₂ COOH)-

-NH-CH(-CH ₂ CH ₂ COOH)-

-NH-CH(-CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ )-

-NH-CH ₂ CH ₂ -

-NH-CH ₂ -O-CH ₂ -

-NH-CH ₂ CH ₂ -O-

-NH-CH ₂ CH ₂ -O-CH ₂ -

-NH-CH ₂ CH ₂ C(-Me) ₂ -

-NH-CH ₂ CH ₂ NH-

-NH-CH ₂ CH ₂ NH-CH ₂ -

-NH-CH ₂ CH ₂ NMe-CH ₂ -

-NH-CH ₂ CH ₂ NH-CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ NMe-CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ N(-CH ₂ COOH)-CH ₂ -

-NH-CH ₂ CH ₂ N(-CH ₂ CH ₂ OH)-CH ₂ -

-NH-CH ₂ CH ₂ N(-CH ₂ CH ₂ OH)-CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ CH ₂ C(=O)-NHCH(-CH ₂ OH)-

-NH-CH ₂ CH ₂ CH ₂ C(=O)-NHCH(-CH ₂ COOH)-

-NH-CH ₂ CH ₂ CH ₂ C(=O)-NHCH(-CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ )-

-NH-CH ₂ CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -

-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH(NH ₂ )-, etc.

In some embodiments, where S* indicates a bond and the coupling agent indicates -Z-A-RL- or -Z-A-RL-Y-.

In some embodiments, where S* indicates a separating agent and the coupling agent indicates -Z-A-S*-RL- or -Z-A-S*-RL-Y-. For example, a dosing agent can be used to mask the hydrophobicity of a drug or other coupling components.

A typical separating agent may include a polyethyleneglycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide, and a dendritic polymer. When the coupling agent contains a polyethyleneglycol (PEG) unit, a cyclodextrin unit, a polyamide, a hydrophilic peptide, a polysaccharide, or a dendritic polymer, these groups may exist as "linear" components or as side chains or branched components.

In some embodiments, where S* incorporates a PEG unit.

In some embodiments, S* combines the following formula:

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ C(=O)-;

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ C(=O)NH-(CH ₂ CH ₂ O)-CH ₂ CH ₂ C(=O)-; or

-NH-(CH ₂ CH ₂ O) _b -CH ₂ CH ₂ NH-(CH ₂ CH ₂ O)-CH ₂ CH ₂ C(=O)-;

where the left and right sides of S* are associated with A and RL, respectively, and b is any integer from 2 to 20. For example, b could be 2, 4, 8, or 12. In some embodiments, the index b is 2. In some embodiments, the index b is 4. In some embodiments, the index b is 8. In some embodiments, the index b is 12.

In some embodiments, the binding agent formulation may be selected from the following structure:

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-;

-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

-C(=O)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-;

Where,

(succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG), or Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG).

In another example, a binding agent may contain the following structure:

Binding agent payload

On the other hand, the present patent application provides a binding agent preparation that contains the binding agent described in this application.

In some embodiments, the binding agent formulation is selected from the following structure: Z'-A-S*-RL-D and Z'-A-S*-RL-Y-D; where Z' indicates a subclone precursor, and the letter A indicates a linkage or shared unit. S* indicates a linkage or separating agent, and Y indicates a spacer subunit; and the letter D indicates a drug unit.

In some embodiments, where the subclone precursor Z' is selected from (maleimide-N-yl)-, (pyrrolidine-2,5-diketone-N-yl)-, M-CH ₂ -C(=O)-NH-, and HS-(CH ₂ ) _1-5 -C(=O)-, where M indicates halogen.

In some embodiments, where the dosage unit includes a cytotoxic agent.

In some embodiments, where the dosage unit comprises: amanitins, anthracyclines, auristatins, baccatins, calicheamicins, camptothecins, cemadotins, colchicines, colcimides, combretastatins, cryptophycins, discodemolides, duocarmycins, docetaxel, doxorubicin, duocarmycins, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, maytansines, maytansinoids, methotrexate, netropsins, pyrrolo[2,1-c][1,4]benzodiazepines (PBD), puromycins, rhizoxins, SN-38, taxanes, tubulysins, vinca alkaloids or derivatives thereof.

In some embodiments, where the dosage unit comprises a DNA topoisomerase I inhibitor.

In some embodiments, the dosage unit comprises camptothecin or a derivative thereof.

In some embodiments, where the medicinal unit combines the following structure:

Dxd

the nitrogen atom of the amino group at position 1 is the group in the attachment position.

In some embodiments, where the binding agent preparation comprises the following structure:

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Maleimid-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

(Pyrrolidine-2,5-diketone-N-yl)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

M-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd; And

HS-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd;

Where,

(Maleimide-N-yl)- has the following structure: , where the nitrogen atom represents a group in the attachment position;

(Pyrrolidine-2,5-diketone-N-yl)- has the following structure: , where the nitrogen atom represents a group in the attachment position;

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG) or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG);

Dxd contains the following structure: , where the nitrogen atom of the amino group in position 1 represents a group in the position of attachment;

M indicates a halogen atom.

In some embodiments, where the binding agent preparation is selected from the following structure:

;

And

.

Units in ligands

Some embodiments of this patent application include ligand units. The ligand unit (C) is a targeting agent that specifically binds to a target moiety. In some embodiments, the ligand unit specifically and selectively binds to cellular components (binding cells) or other target molecules of interest. The function of the ligand unit is to target the drug unit (such as Dxd) and present the drug unit to specific target cell populations that interact with the ligand unit due to the presence of the target component or molecule of the drug unit, allowing free drugs to subsequently be released into the target cell (equivalent to an intracellular process). The ligand unit C includes, but is not limited to, proteins, polypeptides, and peptides.

Suitable ligand units include, for example, antibodies, such as full-length antibodies and antigen-binding fragments, interferons, lymphokines, hormones, growth factors and colony-stimulating factors, vitamins, nutrient transport molecules (including, but not limited to, transferrins), or any other cell-binding molecules or substances. In some embodiments, the ligand unit (C) is an antibody or a protein-targeting agent that is not an antibody.

In some embodiments, the ligand unit is linked to L (the agent-binding unit), which contains the releasable peptide binding agent. As described above, the conjugate described in this article may also contain other linking components to provide additional space between the drug unit and the ligand unit (e.g., a subclone and, optionally, a combined unit A) or to impart solubility-enhancing properties to the combination (e.g., a resolving agent S*). In some of these embodiments, the ligand unit is linked to the agent-binding unit Z via heteroatoms of the ligand unit. Heteroatoms that may be present in the ligand binding unit include sulfur (in one embodiment, from the sulfhydryl group of the targeting ligand), oxygen (in one embodiment, from the carboxyl or hydroxyl group of the targeting ligand), and nitrogen that is optionally substituted (in one embodiment, from a primary or secondary amino functional group of the targeting ligand or, in another embodiment, from an additionally substituted amide nitrogen). These heteroatoms may be present in the targeting ligand in the natural state of the ligand (e.g., in a naturally occurring antibody) or may be introduced into the targeting ligand by chemical modification or bioengineering.

In one embodiment, the ligand unit comprises a sulfhydryl functional group, which facilitates its binding to the linking unit via the sulfur atom of the sulfhydryl functional group.

In another embodiment, the ligand unit comprises one or more lysine residues capable of reacting with activated esters of the precursor subclone of the camptothecin-binding agent intermediate (such esters include, but are not limited to, N-hydroxy succinimide, pentafluorophenyl, and p-nitrophenyl esters), thereby providing an amide linkage consisting of a nitrogen atom of the ligand unit and a C=O group from the subclone of the binding agent unit.

In some aspects, the ligand unit is capable of forming a covalent bond between the subclone (Z) and the ligand unit corresponding to the targeting ligand by reacting with a reactive functional group on the subclone precursor (Z'). The functional group on Z' that is capable of interacting with the targeting ligand will depend on the properties of the ligand unit. In some embodiments, the reactive group is a maleimide that is present in the subclone prior to attachment to form the ligand unit (i.e., the maleimide moiety of the subclone precursor). Covalent attachment between the ligand unit and the subclone is achieved by reacting the sulfhydryl functional group of the ligand unit with the maleimide functional group of Z', resulting in the formation of a sulfur-substituted succinimide. The sulfidryl functional group may be present in the ligand unit in its natural state (e.g., in naturally occurring residues) or may be introduced into the ligand unit through chemical modification or bioengineering.

In another embodiment, the ligand unit is an antibody, and the sulfhydryl group is formed by the reduction of an interchain disulfide in the antibody. Accordingly, in some embodiments, the binding agent unit is conjugated to cysteine residues derived from reduced interchain disulfides.

In another embodiment, the ligand unit is an antibody, and the sulfhydryl functional group is chemically introduced into the antibody, for example, by introducing cysteine residues. Accordingly, in some embodiments, the binding agent unit (with or without an attached drug unit) is conjugated to the ligand unit via cysteine residues introduced by the ligand unit.

In some embodiments, the ligand targets the cell surface or a tumor-associated antigen.

In certain embodiments, the described cell surface receptor or tumor-associated antigen may include HER2, HER3, claudin18.2, folate receptor α (FRα), BCMA, PSMA, TROP-2, CD19, CD20, CD22, CD30, CD79b, EGFR, c-Met, or CEACAM5.

Antibody-drug conjugate (ADC)

On the other hand, the present application provides an antibody-drug conjugate (ADC), which has the following structure: Ab-(LD) _m , where Ab denotes an antibody or an antigen-binding fragment thereof, L denotes a binding agent, D denotes a unit of drug, and m denotes any number from 1 to 10.

In some embodiments, the antibody comprises a monoclonal antibody, a polyclonal antibody, a dimer, a polymer, a multispecific antibody, an intact antibody, an antibody fragment, a human antibody, a humanized antibody, or a chimeric antibody.

In some embodiments, the antigen-binding fragment comprises a Fab, Fab', Fv fragment, F(ab')2, scFv, di-scFv and/or dAb.

In some embodiments, where the antibody is monoclonal.

In some embodiments, the antibody comprises a human antibody, a humanized antibody, or a chimeric antibody.

In some embodiments, where the dosage unit includes a cytotoxic agent.

For example, the drug unit may be a microtubule-interfering agent such as auristatins, such as monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), and auristatin F (AF). Alternatively, the drug may be a microtubule-disrupting agent, such as maytansinoids, such as DM1, DM3, and DM4. Alternatively, the drug unit may be a DNA-damaging agent such as calicheamicins, duocarmycins, SN-38, and pyrrolo[2,1-c][1,4]benzodiazepines (PBD). Another example is the following: the dosage unit may be amanitins, anthracyclines, baccatins, camptothecins, cemadotins, colchicines, colcimides, combretastatins, cryptophycins, dyskermolides, docetaxel, doxorubicin, echinomycins, eleutherobins, epothilones, estramustines, lexitropsins, methotrexes, netropsins, puromycins, rhizoxins, taxanes, tubulysins, or vinca alkaloids.

In some embodiments, the described dosage unit is a topoisomerase-I inhibitor, such as a camptothecin derivative DXd.

In some embodiments, wherein the antibody-drug conjugate is selected from the following structure:

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-(succinimide-3-yl-N)-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-VASSGFG-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-CH ₂ -C(=O)-NH-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ -O-CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ; And

Ab-[-S-CH ₂ CH ₂ -C(=O)-NH-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ O-CH ₂ CH ₂ -C(=O)-RL-NH-CH ₂ CH ₂ CH ₂ -C(=O)-Dxd] _m ;

Where,

(succinimide-3-yl-N)- includes the following structure: , where the dash at position 3 of the structure indicates the position of attachment to the ligand unit, and the dash at position 1 on the N atom indicates the position of attachment to A.

-RL- indicates the following structure: -Val-Ala-Gly-Phe-Gly-(VAGFG), -Val-Ala-Ser-Gly-Phe-Gly-(VASGFG) or -Val-Ala-Ser-Ser-Gly-Phe-Gly-(VASSGFG);

Dxd contains the following structure: , where the nitrogen atom of the amino group in position 1 represents a group in the position of attachment;

Ab denotes an antibody or its antigen-binding fragment, and m denotes any number from 1 to 10.

In some embodiments, wherein the antibody-drug conjugate is selected from the following structure:

;

And

;

where Ab denotes an antibody or its antigen-binding fragment, and m denotes any number from 1 to 10.

In some embodiments, the antibody or antigen-binding fragment thereof comprises: an anti-HER2 antibody, an anti-HER3 antibody, an anti-claudin18.2 antibody, an anti-FRα antibody, or fragments that bind its antigens.

The disclosed antibody-drug conjugate (ADC) compound includes an ADC compound that exhibits anticancer activity. In particular, the ADC compound includes an antibody (i.e., covalently linked) or an antigen-binding fragment (including its antigen-binding fragment) linked to a unit of a drug (such as a cytotoxic agent); for example, the cytotoxic agent exhibits cytotoxicity or growth inhibitory activity when not linked to the antibody or antigen-binding fragment.

The ADC compound of this patent application can selectively deliver an effective dose of a cytotoxic agent or a cytostatic agent to cancer cells or tumor tissues. The ADC described herein has been found to exhibit potent cytotoxicity and/or cell growth inhibitory activity against cells expressing multiple target antigens (e.g., HER2, HER3, claudin18.2, folate receptor α (FRα), CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, or STEAP1). In some embodiments, the cytotoxicity and/or cell growth inhibitory activity of the ADC depends on the expression of the target antigen in the cell. In some embodiments, the described ADC is particularly effective in killing cancer cells expressing target antigens while minimizing the killing of non-target cells. In some embodiments, the described ADC does not exhibit cytotoxicity and/or growth suppression of cancer cells that do not express target antigens.

Variants of cancer types to which the anti-HER3 antibody-drug conjugate according to the present invention can be applied include lung cancer, kidney cancer, urothelial carcinoma, colorectal cancer, prostate cancer, spongioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, metastatic cancer, breast cancer, breast cavity cancer, melanoma, liver cancer, bladder cancer, stomach cancer, gastrointestinal stromal tumor, cervical cancer, head and neck cancer, esophageal cancer, epidermoid carcinoma, peritoneal cancer, glioblastoma multiforme in adults, liver cancer, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal cancer and penile cancer. Treatment using the antibody-drug conjugate against HER3 described in this patent application can target cancer cells in the treatment subject that express the HER3 protein, which is recognized by the antibody of the antibody-drug conjugate. In this specification, the term "HER3-expressing cancer" refers to a cancer that contains cells with the HER3 protein on their surface or to a cancer that secretes the HER3 protein into the bloodstream.

Cancer types to which the anti-claudin18.2 antibody-drug conjugate of this patent application may be applied include gastrointestinal cancer, pancreatic cancer, esophageal cancer, non-small cell lung cancer, liver cancer, ovarian cancer, lung cancer, gallbladder carcinoma, or head and neck cancer.

Cancer types to which the anti-FRα antibody-drug conjugate of the present invention can be applied include ovarian cancer, breast cancer, lung cancer, colorectal cancer, or kidney cancer.

Examples of cancer types to which the HER2 antibody-drug conjugate of the present invention can be applied include, but are not limited to, mast cell carcinoma, gastric cancer, bladder cancer, urothelial carcinoma, esophageal cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., severe endometrial cancer of the uterus), salivary duct carcinoma, cervical cancer, endometrial cancer, and ovarian cancer (English et al. (2013) Mol Diagn Ther. [Molecular Diagnostics and Therapy] 17:85-99).

In this patent application, the antibody may target target cell surface receptors or tumor-associated antigens. Furthermore, antibodies may include full-length antibodies or partially reactive antibody fragments, or complex fragments of antibodies bound to receptors, antigens, or other target cells. An antibody may be any protein, protein-like molecule, or polypeptide that binds to, complexes with, or reacts with a portion of a cell population requiring therapeutic modification. In some embodiments, the described antibody may be a chimeric antibody or a functionally active fragment thereof, a humanized antibody or a functionally active fragment thereof, or a human antibody or a functionally active fragment thereof.

In some embodiments, the antibody may also be a functionally active fragment, derivative, or analog of an antibody, wherein the antibody can bind to a target antigen using immunological specificity (e.g., a cancer antigen, a viral antigen, a microbial antigen, or another antigen that can bind to cells or a matrix). Here, "functionally active" means that the described fragment, derivative, or analog can recognize the same antigen and recognize a fragment, derivative, or analog antibody derived from the antigen. Other useful antibodies include antibody fragments such as, but not limited to, F(ab')2 fragments, Fab fragments, Fab' fragments, Fv fragments, and heavy-light chain dimers of an antibody, or Fv fragments, or single-chain antibodies (scFv). In other embodiments, the antibody may be a fusion protein or a functionally active fragment thereof.

The antibodies of this patent application may include, but are not limited to, antibodies against the following antigens (examples of cancer are given in parentheses): CA125 (ovarian cancer), CA15-3 (cancer, carcinoma), CA19-9 (cancer), L6 (cancer), Lewis Y (cancer), Lewis X (cancer), alpha-fetal protein (cancer), CA 242 (colorectal cancer), placental alkaline phosphatase (cancer), prostate cancer-specific membrane antigen (prostate cancer), prostate acid phosphatase (prostate cancer), epidermal growth factor (cancer), MAGE-1 (cancer), MAGE-2 (cancer), MAGE-3 (cancer), MAGE-4 (cancer), antitransferrin receptor (cancer), p97 (melanoma), MUC1-KLH (mastocarcinoma), CEA (colorectal cancer), gp100 (melanoma), MART1 (melanoma), Prostate-specific antibody (PSA) (prostate cancer), IL-2 receptor (T-cell leukemia and lymphoma), CD20 (non-Hodgkin lymphoma), CD52 (leukemia), CD33 (leukemia), CD22 (lymphoma), human chorionic gonadotropin (cancer), CD38 (multiple myeloma), CD40 (lymphoma), mucin (cancer), P21 (cancer), MPG (melanoma), and Neu oncogene product (cancer).

The antibodies of this patent application may also include modified or unmodified analogs and derivatives (i.e., by covalent attachment of any molecule), provided that such covalent attachment allows the antibody to retain its antigen-binding immunospecificity. Embodiments include, but are not limited to, analogs and derivatives of antibodies, including additional modifications such as glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization via acquired protecting/blocking groups, protease cleavage, attachment to a cellular antibody unit or to other proteins, etc. Any number of chemical modifications may be introduced using acquired methods, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc. In addition, analogs or derivatives may include one or more unnatural amino acids. In some embodiments, the antibody may contain alterations (e.g., substitution, deletion, or addition) of amino acid residues that interact with the Fc receptor.

In some embodiments, the antibody or antigen-binding fragment thereof comprises: an anti-HER2 antibody, an anti-claudin18.2 antibody, an anti-FRα antibody, an anti-HER3 antibody, or fragments that bind its antigens.

In one embodiment, the antibody is an anti-HER3 antibody, wherein the anti-HER3 antibody comprises a VH comprising HCDR1, HCDR2 and HCDR3, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 1. HCDR2 comprises the amino acid sequence of SEQ ID NO: 2. HCDR3 comprises the amino acid sequence of SEQ ID NO: 3.

In some embodiments, the HER3 antibody comprises a VH, wherein the VH comprises the amino acid sequence of SEQ ID NO: 15.

In some embodiments, the anti-HER3 antibody comprises a heavy chain constant region of the antibody. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG1, IgG2, IgG3, or IgG4.

In some embodiments, the anti-HER3 antibody comprises an antibody heavy chain. The antibody heavy chain comprises the amino acid sequence of SEQ ID NO: 17.

In some embodiments, wherein the anti-HER3 antibody comprises a VL, wherein the VL comprises LCDR1, LCDR2, and LCDR3. LCDR1 comprises the amino acid sequence of SEQ ID NO: 4. LCDR2 comprises the amino acid sequence of SEQ ID NO: 5. LCDR3 comprises the amino acid sequence of SEQ ID NO: 6.

In some embodiments, wherein the anti-HER3 antibody comprises VH and VL, wherein VH comprises HCDR1, HCDR2 and HCDR3, and VL comprises LCDR1, LCDR2 and LCDR3; wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 1. HCDR2 comprises the amino acid sequence of SEQ ID NO: 2. HCDR3 comprises the amino acid sequence of SEQ ID NO: 3. LCDR1 comprises the amino acid sequence of SEQ ID NO: 4. LCDR2 comprises the amino acid sequence of SEQ ID NO: 5. LCDR3 comprises the amino acid sequence of SEQ ID NO: 6.

In some embodiments, wherein the anti-HER3 comprises a VL, wherein the VL comprises the amino acid sequence of SEQ ID NO: 16.

In some embodiments, wherein the anti-HER3 comprises VH and VL, wherein VH comprises the amino acid sequence of SEQ ID NO: 15, VL comprises the amino acid sequence of SEQ ID NO: 16.

In some embodiments, the anti-HER3 antibody comprises the constant region of a light chain antibody. For example, the light chain constant region of the antibody may comprise the constant region of human Igκ or human Igλ.

In some embodiments, the HER3 antibody comprises a light chain. The light chain comprises the amino acid sequence of SEQ ID NO: 18.

In some embodiments, the HER3 antibody comprises a heavy chain. The heavy chain comprises the amino acid sequence of SEQ ID NO: 17. The light chain comprises the amino acid sequence of SEQ ID NO: 18.

In one embodiment, where the antibody is an anti-Claudin18.2 antibody, where the anti-Claudin18.2 antibody comprises a VH, where the VH comprises HCDR1, HCDR2 and HCDR3, HCDR1 comprises the amino acid sequence of SEQ ID NO: 19. Wherein HCDR2 comprises the amino acid sequence of SEQ ID NO: 20. HCDR3 comprises the amino acid sequence of SEQ ID NO: 21.

In some embodiments, wherein the anti-Claudin18.2 antibody comprises a VH, wherein the VH comprises the amino acid sequence of SEQ ID NO: 32.

In some embodiments, the anti-Claudin18.2 antibody comprises a heavy chain constant region of an antibody. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG1, IgG2, IgG3, or IgG4.

In some embodiments, the anti-Claudin18.2 antibody comprises an antibody heavy chain. The antibody heavy chain comprises the amino acid sequence of SEQ ID NO: 34.

In some embodiments, wherein the anti-Claudin18.2 antibody comprises a VL, wherein the VL comprises LCDR1, LCDR2 and LCDR3, LCDR1 comprises the amino acid sequence of SEQ ID NO: 22. LCDR2 comprises the amino acid sequence of SEQ ID NO: 23. LCDR3 comprises the amino acid sequence of SEQ ID NO: 24.

In some embodiments, wherein the anti-Claudin18.2 antibody comprises VH and VL, wherein VH comprises HCDR1, HCDR2 and HCDR3, and VL comprises LCDR1, LCDR2 and LCDR3; wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 19. HCDR2 comprises the amino acid sequence of SEQ ID NO: 20. HCDR3 comprises the amino acid sequence of SEQ ID NO: 21. LCDR1 comprises the amino acid sequence of SEQ ID NO: 22. LCDR2 comprises the amino acid sequence of SEQ ID NO: 23. LCDR3 comprises the amino acid sequence of SEQ ID NO: 24.

In some embodiments, wherein the anti-Claudin18.2 antibody comprises a VL, wherein the VL comprises the amino acid sequence of SEQ ID NO: 33.

In some embodiments, wherein the anti-Claudin18.2 antibody comprises VH and VL, wherein VH comprises the amino acid sequence of SEQ ID NO: 32, VL comprises the amino acid sequence of SEQ ID NO: 33.

In some embodiments, the anti-Claudin18.2 antibody comprises the constant region of an antibody light chain. For example, the constant region of the antibody light chain may comprise the constant region of human Igκ or human Igλ.

In some embodiments, the anti-Claudin18.2 antibody comprises an antibody light chain. The antibody light chain comprises the amino acid sequence of SEQ ID NO: 35.

In some embodiments, the anti-Claudin18.2 antibody comprises an antibody heavy chain. The antibody heavy chain comprises the amino acid sequence of SEQ ID NO: 34. The antibody light chain comprises the amino acid sequence of SEQ ID NO: 35.

In one embodiment, where the antibody is an anti-FRα antibody, where the anti-FRα antibody comprises a VH, where the VH comprises HCDR1, HCDR2 and HCDR3, HCDR1 comprises the amino acid sequence of SEQ ID NO: 36, and HCDR2 comprises the amino acid sequence of SEQ ID NO: 37. And HCDR3 comprises the amino acid sequence of SEQ ID NO: 38.

In some embodiments, wherein the anti-FRα antibody comprises a VH, wherein the VH comprises the amino acid sequence of SEQ ID NO: 49.

In some embodiments, the anti-FRα antibody comprises a heavy chain constant region of an antibody. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG. For example, the heavy chain constant region of the antibody may comprise a constant region derived from human IgG1, IgG2, IgG3, or IgG4.

In some embodiments, the anti-FRα antibody comprises an antibody heavy chain. The antibody heavy chain comprises the amino acid sequence of SEQ ID NO: 51.

In some embodiments, wherein the anti-FRα antibody comprises a VL, wherein the VL comprises LCDR1, LCDR2 and LCDR3, LCDR1 comprises the amino acid sequence of SEQ ID NO: 39. LCDR2 comprises the amino acid sequence of SEQ ID NO: 40. LCDR3 comprises the amino acid sequence of SEQ ID NO: 41.

In some embodiments, wherein the anti-FRα antibody comprises VH and VL, wherein VH comprises HCDR1, HCDR2 and HCDR3, and VL comprises LCDR1, LCDR2 and LCDR3; wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 36. HCDR2 comprises the amino acid sequence of SEQ ID NO: 37. HCDR3 comprises the amino acid sequence of SEQ ID NO: 38. LCDR1 comprises the amino acid sequence of SEQ ID NO: 39. LCDR2 comprises the amino acid sequence of SEQ ID NO: 40. LCDR3 comprises the amino acid sequence of SEQ ID NO: 41.

In some embodiments, wherein the anti-FRα antibody comprises a VL, wherein the VL comprises the amino acid sequence of SEQ ID NO: 50.

In some embodiments, wherein the anti-FRα antibody comprises VH and VL, wherein VH comprises the amino acid sequence of SEQ ID NO: 49, VL comprises the amino acid sequence of SEQ ID NO: 50.

In some embodiments, the anti-FRα antibody comprises the constant region of an antibody light chain. For example, the constant region of an antibody light chain may comprise the constant region of human Igκ or human Igλ.

In some embodiments, the anti-FRα antibody comprises an antibody light chain. The antibody light chain comprises the amino acid sequence of SEQ ID NO: 52.

In some embodiments, the anti-FRα antibody comprises an antibody heavy chain. The antibody heavy chain comprises the amino acid sequence of SEQ ID NO: 51. The antibody light chain comprises the amino acid sequence of SEQ ID NO: 52.

Compound

On the other hand, the present patent application proposes a combination of drugs that comprises a conjugate described in this patent application and an additional carrier that is pharmaceutically acceptable.

Typically, the term "carrier" refers to a diluent, adjuvant, or excipient administered with a compound. Such a drug carrier may be in liquid form, such as water or an oil, including petroleum, animal oil, vegetable oil, or synthetic oil such as peanut oil, soybean oil, mineral oil, and sesame oil. The carrier may also be saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silicon dioxide, or urea. It may also be an adjuvant, stabilizer, thickener, lubricant, or colorant. In one embodiment, when the drug is administered to a patient, the pharmaceutically acceptable chemical or composition and carrier are sterile.

The conjugate according to this patent application may be in any form that allows the compound (dosage unit) to be administered to a patient for the treatment of a symptom related to the expression of an antigen bound to the ligand unit. For example, the conjugate may be in liquid or solid form. An example of a route of administration is parenteral administration. Parenteral administration includes subcutaneous, intravenous, intramuscular, intrasternal injection, or intravenous drip. In one embodiment, the composition is administered parenterally. In another embodiment, the composition is administered intravenously. It can be administered by any convenient route, such as intravenous drip or bolus infusion.

Materials used in the preparation of pharmaceutical formulations may be nontoxic in the amounts used. Those skilled in the art readily recognize that the optimal dosage of one or more active ingredients in a pharmaceutical formulation depends on various factors. Relevant factors include, but are not limited to, the type of mammal (e.g., human), the specific form of the compound, the route of administration, and the formulation used.

For example, the composition may be in liquid form. The liquid can be administered by injection. The composition for injection may include one or more surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers, and isotonic agents. When the composition is administered intravenously, an example of a carrier can be water. Saline solution, an aqueous dextrose-glycerol solution, and water can also be used as liquid carriers, especially injectable solutions. Suitable drug carriers also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dry skim milk, glycerol, propylene, ethylene glycol, water, and ethanol. If necessary, the composition of the present invention may also contain a small amount of a humectant, emulsifier or pH buffer.

The pharmaceutical formulation can be prepared so that the compound is bioavailable after administration to the patient. The formulation may be in the form of single or multiple dosage units. For intravenous administration, the formulation may contain from approximately 0.01 mg to approximately 100 mg of the conjugate per kilogram of patient body weight. The dose may be even lower depending on the drug used, for example, from 1.0 mcg/kg to 5.0 mg/kg, 4.0 mg/kg, 3.0 mg/kg, 2.0 mg/kg, or 1.0 mg/kg, or from 1.0 mcg/kg to 500.0 mcg/kg of patient body weight.

Typically, the dose of the conjugate administered to the patient is from about 0.01 mg/kg to about 100 mg/kg of the patient's body weight, or from 1.0 μg/kg to 5.0 mg/kg of the patient's body weight. In some embodiments, the dose of the conjugate administered to the patient is from about 0.01 mg/kg to about 15 mg/kg of the patient's body weight. In some embodiments, the dose of the conjugate administered to the patient is from about 0.1 mg/kg to about 15 mg/kg of the patient's body weight. In some embodiments, the dose of the conjugate administered to the patient is from about 0.1 mg/kg to about 20 mg/kg of the patient's body weight. In some embodiments, the dose of the conjugate administered to the patient is from about 0.1 mg/kg to about 5 mg/kg of the patient's body weight, or from about 0.1 mg/kg to about 10 mg/kg of the patient's body weight.

Target

On the other hand, the present patent application provides for the use of a conjugate or combination of drugs as described in this patent application in the preparation of a medicament for the treatment of diseases or disease states.

On the other hand, the present patent application provides a method for treating diseases or disease states in patients in need thereof, comprising administering to the patients an effective dose of the conjugate as described in this application.

In some embodiments, where the diseases or conditions include tumors.

Cancers that can be treated with the ADCs of this patent application include, but are not limited to, cancers of the hematopoietic system, such as lymphomas (Hodgkin's lymphoma and non-Hodgkin's lymphoma), as well as leukemia and solid tumors. Cancers of the hematopoietic system include follicular lymphoma, anaplastic large cell lymphoma, mantle cell lymphoma, acute myeloid leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, and multiple myeloma. Solid tumors include fibrosarcoma, myxosarcomas, liposarcomas, chondrosarcomas, osteosarcomas, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma. In addition, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer, oral cancer, rhinocarcinoma, throat cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hidradoma carcinoma, sebaceous carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, liver cancer, bile duct cancer, choriocarcinoma, seminoma, embryonal cancer, Wilms' tumor, cervical cancer, uterine cancer, testicular carcinoma, small cell lung cancer, bladder cancer, lung cancer, epicytoma, glioma, glioblastoma multiforme, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineloma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, skin cancer, melanoma, neuroblastoma and retinoblastoma.

In some embodiments, wherein the tumors include tumors with abnormal expression of HER3.

In some embodiments, wherein the tumors include tumors with high HER3 expression.

In some embodiments, wherein the tumors include tumors with abnormal expression of claudin18.2.

In some embodiments, tumors with high expression of claudin18.2 are included.

In some embodiments, tumors with abnormal expression of FRα are included.

In some embodiments, tumors with high FRα expression are included.

In order not to be limited by any single theory, the following embodiments are intended only to illustrate the binding agents, drug binding agents, conjugates, methods of preparation, purposes, etc. of this application, but are not intended to limit the scope of the application.

Execution option

Option 1

1.1 ADC construction method

Replace the anti-HER3 antibody, anti-Claudin18.2 antibody, and anti-folate receptor FRα antibody in the target buffer (citrate buffer or histidine hydrochloride buffer, pH 6.0-6.5) using a replacement device (desalting column, ultrafiltration device, etc.), and then measure the antibody concentration. Maintain the antibody concentration at 5-20 mg/mL and incubate it with 0.04-0.4 mM TCEP (reducing agent) for 1-5 hours at 10-37°C to completely open the disulfide bond between the antibody chains. Next, adjust the pH of the reconstituted antibody solution to 6.0 with phosphoric acid and lower its temperature to room temperature. Dissolve the small molecule binding agent in DMSO and calculate its concentration. Mix the solution of the coupling agent and the drug (04-Dxd, 07-Dxd, 08-Dxd, 09-Dxd, where the amino acid unit in 04-Dxd is replaced by GGFG) with the fully reduced antibody solution according to the corresponding drug. Maintain the molecular ratio of the antibody (molar ratio > 10) and maintain the DMSO ratio below 10% (v/v) for 0.5-2 h until the reaction is complete. Filter the crude ADC solution, purify it with a purifying device (desalting column, ultrafiltration device, tangential flow system, etc.) to remove small molecular organic impurities and excess DMSO from the solution, and then place the ADC product in a preservation buffer.

Table 1. Sequence of the light chain and heavy chain of the antibody

TYPE NAME SEQUENCE IDENTIFIER NO. Antibody against HER3 RS-HT-002 HC: SEQ ID NO: 17
LC: SEQ ID NO: 18 Antibody against Claudin18.2 hu782 HC: SEQ ID NO: 34
LC: SEQ ID NO: 35 Antibodies to the folate receptor FRα M9346A HC: SEQ ID NO: 51
LC: SEQ ID NO: 52

Table 2. Drug structure of the binding agent

Drug binding agent structure 04-Dxd 07-Dxd 08-Dxd 09-Dxd

1.2 Detection of DAR of ADC product

Detected using the HIC-HPLC method. Collect a sample and extract it at pH 7.0 under mobile phase conditions. Determine the peak absorbance area at the specified wavelength. Specific detection methods are listed in Table 3. The DAR of the ADC product should be 7-8.

Table 3: HIC-HPLC detection method

Column: Hydrophobic Interaction Chromatography Column (HIC) Consumption: 0.5-1 ml/min. Stop time: 15-30 min. Injection quantity: 10-20 micrograms Mobile phases: Mobile phase A: phosphate buffer + high concentration ammonium sulfate, pH=7.0±0.2
Mobile phase B: Phosphate buffer + 15-25% v/v isopropyl alcohol, pH = 7.0 ± 0.2.

1.3 DAR HIC-HPLC Detection Results

HER3-ADC Claudin18.2-ADC FRα-ADC HER3-ADC04 HER3-ADC07 HER3-ADC08 HER3-ADC09 Claudin18.2-ADC08 Claudin18.2-ADC09 FRα-ADC08 FRα-ADC09 DAR 8.05 7.97 7.93 7.97 7.67 7.42 8.00 8.00

Option 2

2.1 In vitro assessment of HER3 ADC activity

Treat SW620 human colon cancer cells in logarithmic phase with pancreatin, resuspend them in the appropriate medium, adjust the density to 2×104/mL, add 100 μL/well to a 96-well microwell plate to achieve a cell count of 2× ¹⁰³ cells per well, then add 200 μL of medium to each well. Culture at 37°C and 5% _CO2 for 48 hours, then remove the supernatant from the cell culture plate, dilute the binding agent - small molecule drugs (04-Dxd, 07-Dxd, 08-Dxd, 09-Dxd), HER3 ADC, drugs (HER3-ADC04, HER3-ADC07, HER3-ADC08, HER3-ADC09) and carry out the interaction control of monoclonal antibodies with the medium with 400 nM as the starting concentration for the double gradient dilution: a total of 9 concentration gradients. Then add 100 μl of the diluted sample to one well. Culture for 4-6 additional days to allow color development, then remove the plate with cells and let it sit at room temperature for 15-20 minutes, add 100 µl of CellTiter-Glo® Luminescent Cell Viability Assay Solution (promega), shake oscillatingly at 200 rpm for 2 minutes, then keep in the dark for 18 minutes and count with an M5 microplate reader.

Processing the result:

then perform a four-parameter curve fit with the logarithm of the concentration as the horizontal coordinate and the kill rate as the vertical coordinate.

Result Figure 2A shows that, compared with naked antibody (RS-HT-002), Dxd-conjugated HER3 monoclonal antibody has a more obvious killing effect on SW620 target cells.

Figure 2B shows that all the -Dxd binding agents have killing effects on SW620 human colon cancer cells, and the killing effects of 07-Dxd, 08-Dxd, and 09-Dxd are obviously more effective than that of 04-Dxd.

Figure 3 shows that all the anti-HER3 ADC antibodies containing different binding agents have killing effects on SW620 human colon cancer cells, and the killing effect of HER3-ADC07, HER3-ADC08 and HER3-ADC09 is apparently more effective than that of HER3-ADC04.

Figure 4 shows that all HER3 ADC antibodies containing different binding agents have no obvious killing effect on non-target CHO-K1 cell.

2.2 Evaluation of HER3 ADC plasma stability

To determine the stability of ADCs in vitro, this study utilized in vitro human plasma incubation to simulate the in vivo environment for detection. Specifically, HER3 ADC was added to 10% human plasma to achieve a final antibody concentration of 1 mg/mL, followed by incubation at 37°C for 3 and 7 days, and then cell death detection. Target SW620 cells were treated in logarithmic phase, plated at a specific density, cultured at 37°C and 5% _CO2 for 48 hours, and then incubated ADC gradient-diluted with plasma was added. Unincubated ADC diluted with plasma served as a control for the experiment.

Result: Figure 5 shows that HER3-ADC08 and HER3-ADC09 have no obvious change in cell killing activity after 3 days or 7 days incubation in plasma.

2.3 Evaluation of HER3 ADC activity in vivo

Select 4-6-week-old female SPF BALB/c nude mice and feed them adoptively for 3-4 days for tumor inoculation. Treat log-phase SW620 human colon cancer cells, wash twice with 1640 serum-free medium, dilute to 2.5× ¹⁰⁷ /mL, and then inject 0.2 mL of 5× ¹⁰⁶ cells per mouse subcutaneously into the right axilla. When the tumor increases to 150±50 ^mm3 within one week, randomly group 5 mice in each group. Take the blank model group with 2, 6, and 10 mg/kg ADCs as the PBS control. Inject the small molecule control group with the same molar mass of ADCs at 10 mg/kg once a week. Doses of 6 and 10 mg/kg are administered four times, and the 2 mg/kg dose is administered eight times, all administered intraperitoneally. Tumors are measured twice a week and total tumor volume and mouse body weight are recorded before each administration.

The total tumor volume can be calculated using the following formula: V( ^mm3 )=( ^W2 ×L)/2, where W is the short diameter of the tumor midline, and L is the longitudinal diameter. If the total tumor volume is greater than 2000 ^mm3 and the mass loss exceeds 20%, or if disruption of vital physiological functions or necrosis of tumor ulcers is detected, mice are euthanized by cervical dislocation.

Result: Figure 6 shows that HER3-ADC07, HER3-ADC08, and HER3-ADC09 can significantly suppress the growth of transplanted SW620 tumors. In addition, tumors in mice completely regressed after four doses of 6 and 10 mg/kg. Tumors in some mice began to grow three weeks after stopping treatment. A dose of 2 mg/kg was able to suppress tumor growth in all mice. HER3-ADC08 was superior to HER3-ADC07 and HER3-ADC09 in tumor suppression. Figure 7 shows that there was no significant difference in body weight between mice in the ADC group and the PBS control group during treatment.

Option 3

3.1 In vitro assessment of Claudin18.2 ADC activity

Treat log-phase HEK293-hCLDN18.2, NUGC4, and NUGC4-hCLDN18.2 with pancreatin, resuspend them in the appropriate medium, adjust the density to 2× ¹⁰⁴ /mL, add 100 µL/well to a 96-well microwell plate to achieve 2× ¹⁰³ cells per well, then add 200 µL of medium to each well. Cultivate at 37°C and 5% _CO2 for 48 hours, then remove the supernatant from the cell culture plate, dilute the binding agent - small molecule drugs (08-Dxd and 09-Dxd), ADC drugs (Claudin18.2-ADC08, Claudin18.2-ADC09) and control monoclonal antibody (hu782) with the medium with 400 nM as the starting concentration for a double gradient dilution, a total of 9 concentration gradients. Add 100 μl of the diluted sample to one well. Culture for 4-6 additional days to allow color development, then remove the plate with cells and let it sit at room temperature for 15-20 minutes, add 100 µl of CellTiter-Glo® Luminescent Cell Viability Assay Solution (promega), shake oscillatingly at 200 rpm for 2 minutes, then keep in the dark for 18 minutes and count with an M5 microplate reader.

Processing the result:

then perform a four-parameter curve fit with the logarithm of the concentration as the horizontal coordinate and the kill rate as the vertical coordinate.

Result: Figures 8, 9 and 10 showed that the Claudin18.2-ADC08 and Claudin18.2-ADC09 antibody conjugates had obvious killing effects on NUGC4, NUGC4-hCLDn18.2 and HEK293-hCLDn18.2 and Claudin18.2-positive target cells.

3.2 Plasma stability assessment of Claudin18.2 ADC

To detect the stability of ADC preparations in vitro, this study utilizes in vitro incubation of human plasma to simulate the in vivo environment for detection. Specifically, Claudin18.2-ADC08 is added to 10% human plasma to achieve a final antibody concentration of 1 mg/mL, followed by incubation at 37°C for 3 and 7 days, and then cell death detection is performed. Target HEK293-hCLDN18.2 cells are incubated in logarithmic phase, plated at a certain density, cultured at 37°C and 5% _CO2 for 48 hours, and the incubated Claudin18.2-ADC08 gradiently diluted with plasma is added. Unincubated Claudin18.2-ADC08 diluted with plasma is used as a control for the experiment.

Result Figure 11 shows that Claudin18.2-ADC08 has no obvious change in cell killing activity after 3 days or 7 days incubation in plasma.

3.3 Evaluation of Claudin18.2 ADC activity in vivo

Select 4-6-week-old female SPF BALB/c nude mice and feed them adoptively for 3-4 days for tumor inoculation. Treat log-phase NUGC4 cells, wash twice with 1640 serum-free medium, dilute to 2.5× ¹⁰⁷ /mL, and then inject 0.2 mL of 5× ¹⁰⁶ cells per mouse subcutaneously into the right axilla. When the tumor grows to 150±50 ^mm3 within one week, randomly group 5 mice into each group. Use the blank group as a PBS control. Inject a low-molecular-weight control group with the same molar mass as the ADC drugs at a dose of 10 mg/kg, and administer Claudin18.2-ADC08 intraperitoneally at doses of 1 mg/kg, 3 mg/kg, and 10 mg/kg once weekly four times. Continue to measure tumors twice weekly after dosing and record total tumor volume and mouse body weight before each administration.

The total tumor volume can be calculated using the following formula: V( ^mm3 )=( ^W2 ×L)/2, where W is the short diameter of the tumor midline, and L is the longitudinal diameter. If the total tumor volume is greater than 2000 ^mm3 and the mass loss exceeds 20%, or if disruption of vital physiological functions or necrosis of tumor ulcers is detected, mice are euthanized by cervical dislocation.

Result: Figure 12 showed that the NUGC4 Claudin18.2-ADC08 mouse subcutaneous transplantation tumor model had antitumor activity and a certain dose response.

Option 4

4.1 Evaluation of the activity of the FR α ADC drug of the folate receptor in laboratory conditions

Treat log phase OVCAR3 ovarian cancer cells with pancreatin, resuspend them in the corresponding medium, adjust the density to 2× ¹⁰⁴ /mL, add 100 μL/well to a 96-well microwell plate to achieve 2× ¹⁰³ cells/well, then add 200 μL of medium to each well. Grow at 37°C and 5% _CO2 for 48 hours, then remove the supernatant from the cell culture plate, dilute the binding agent - small molecule drugs (08-Dxd and 09-Dxd), ADC drugs (FRα-ADC08 and FRα-ADC09), and control monoclonal antibody (M9346A) with the medium with 400 nM as the starting concentration for double gradient dilution, a total of 9 concentration gradients. Add 100 µl of the diluted sample to one well. Incubate for an additional 4-6 days to allow color development, then remove the plate and let it sit at room temperature for 15-20 minutes. Add 100 µl of CellTiter-Glo® Luminescent Cell Viability Assay Solution (promega), shake oscillatingly at 200 rpm for 2 minutes, then incubate in the dark for 18 minutes and count with an M5 microplate reader.

Processing the result:

then perform a four-parameter curve fit with the logarithm of the concentration as the horizontal coordinate and the kill rate as the vertical coordinate.

Result: Figure 13 shows that the FRα antibody conjugates FRα-ADC08 and FRα-ADC09 have significant killing activity against OVCAR3 cells.

4.2 Evaluation of folate receptor stability of FR α ADC in plasma

To detect the stability of ADC preparations in vitro, this study utilizes in vitro incubation of human plasma to simulate the in vivo environment for detection. Specifically, FRα-ADC08 is added to 50% human plasma to achieve a final antibody concentration of 1 mg/mL, followed by incubation at 37°C for 3 and 7 days, and then cell death detection. Target cells or OVCAR3 target cells are inoculated in the logarithmic phase, plated at a specific density, and cultured at 37°C and 5% _CO2 for 48 hours. The incubated FRα-ADC08 gradient diluted with plasma is then added, and unincubated FRα-ADC08 diluted with plasma is used as a control for the experiment.

Result Figure 14 shows that FRα-ADC08 has no obvious change in cell killing activity after 3 days or 7 days incubation in plasma.

4.3 Evaluation of the activity of the folate receptor FR α ADC in a living organism

Select 3-4-week-old female SPF BALB/c nude mice and feed them adoptively for 3-4 days for tumor inoculation. Treat the mice with OVCAR3 in log phase, wash twice with 1640 serum-free medium, dilute to 3× ¹⁰⁷ /mL and mix with an equal volume of matrix gel, then inoculate the mice subcutaneously in the right axilla at 0.2 mL of 3× ¹⁰⁶ cells per mouse. When the tumor grows to 150±50 ^mm3 within 1 week, randomly group 5 mice in each group. Use the blank group as a PBS control. Inject a control group of small molecules with the same molar mass as the ADCs at a dose of 5 mg/kg, and administer FRα-ADC08 intraperitoneally at doses of 2 mg/kg and 5 mg/kg once weekly. Continue to measure the tumor twice weekly and record the total tumor volume and mouse body weight before each administration.

The total tumor volume can be calculated using the following formula: V( ^mm3 )=( ^W2 ×L)/2, where W is the short diameter of the tumor midline, and L is the longitudinal diameter. If the total tumor volume is greater than 2000 ^mm3 and the mass loss exceeds 20%, or if disruption of vital physiological functions or necrosis of tumor ulcers is detected, mice are euthanized by cervical dislocation.

Result: Figure 15 showed that FRα-ADC08, by inhibiting the subcutaneous tumor transplantation model in OVCAR3 mice, had an inhibitory effect on tumor growth.

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<InventionTitle languageCode="en">ANTIBODY-DRUG CONJUGATE AND USE

THEREOF</InventionTitle>

<SequenceTotalQuantity>52</SequenceTotalQuantity>

<SequenceData sequenceIDNumber="1">

<INSDSeq>

<INSDSeq_length>10</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..10</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q1">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

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<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q105">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GYTFSNYWID</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="2">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q106">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q107">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>EILPGSGGTDYNEKFKG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="3">

<INSDSeq>

<INSDSeq_length>8</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..8</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q108">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..8</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q109">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DDYDVFAY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="4">

<INSDSeq>

<INSDSeq_length>16</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..16</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q110">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..16</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q111">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RSSKSLLHSNGITYLY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="5">

<INSDSeq>

<INSDSeq_length>7</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q112">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q113">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QMSNLAS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="6">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q114">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q115">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>AQNLELPWT</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="7">

<INSDSeq>

<INSDSeq_length>25</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q116">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q117">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVKVSCKAS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="8">

<INSDSeq>

<INSDSeq_length>14</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q118">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q119">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WVRQAPGQGLEWMG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="9">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q120">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q121">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>KATITADTSTSTAYMELSSLRSEDTAVYYCAR</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="10">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q122">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HFR4; anti-Claudin18.2

HFR4</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q123">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WGQGTLVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="11">

<INSDSeq>

<INSDSeq_length>23</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q124">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q125">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPGTLSLSPGERATLSC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="12">

<INSDSeq>

<INSDSeq_length>15</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q126">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q127">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WYQQKPGQAPRLLIY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="13">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q128">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q129">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GVPDRFSSSGSGTDFTLTISRLEPEDFAVYYC</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="14">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q130">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LFR4</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q131">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>FGQGTKVEIKR</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="15">

<INSDSeq>

<INSDSeq_length>117</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..117</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q132">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 VH</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..117</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q133">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVKVSCKASGYTFSNYWIDWVRQAPGQGLEWMG

EILPGSGGGTDYNEKFKGKATITADTSTSTAYMELSSLRSEDTAVYYCARDDYDVFAYWGQGTLVTVSS</

INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="16">

<INSDSeq>

<INSDSeq_length>113</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..113</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q134">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 VL</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..113</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q135">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPGTLLSLPGERATLSCRSSKSLLHSNGITYLYWYQQKPGQAP

RLLIYQMSNLASGVPDRFSSSGSGTDFTLTISRLEPEDFAVYYCAQNLELPWTFGQGTKVEIKR</INSD

Seq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="17">

<INSDSeq>

<INSDSeq_length>447</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..447</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q136">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 HC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..447</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q137">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVKKPGSSVKVSCKASGYTFSNYWIDWVRQAPGQGLEWMG

EILPGSGGGTDYNEKFKGKATITADTSTSTAYMELSSLRSEDTAVYYCARDDYDVFAYWGQGTLVTVSSAS

TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP

SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV

TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP

APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD

SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="18">

<INSDSeq>

<INSDSeq_length>219</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..219</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q138">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-HER3 LC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..219</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q139">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPGTLLSLPGERATLSCRSSKSLLHSNGITYLYWYQQKPGQAP

RLLIYQMSNLASGVPDRFSSSGSGTDFTLTISRLEPEDFAVYYCAQNLELPWTFGQGTKVEIKRTVAAPS

VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA

DYEKHKVYACEVTHQGLSSPVTKSFNRGEC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="19">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q140">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q141">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GYSITSGYSWH</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="20">

<INSDSeq>

<INSDSeq_length>16</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..16</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q142">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..16</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q143">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>YIHYSDNTNYAPSLKS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="21">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q144">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q145">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>IYYGNSFVY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="22">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q146">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q147">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>KSSQSLLNSGNQKNYLT</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="23">

<INSDSeq>

<INSDSeq_length>7</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q148">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q149">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WASTRES</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="24">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q150">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q151">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QNDYFYPYT</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="25">

<INSDSeq>

<INSDSeq_length>25</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q152">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q153">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLQESGPGLVKPSETLSLTCTVS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="26">

<INSDSeq>

<INSDSeq_length>14</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q154">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q155">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WIRQPPGKGLEWMG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="27">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q156">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

HFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q157">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RITISRDTSKNQFSLKLSSVTAADTAVYYCAR</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="28">

<INSDSeq>

<INSDSeq_length>23</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q158">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q159">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPDSLAVSLGERATINC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="29">

<INSDSeq>

<INSDSeq_length>15</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q160">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q161">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WYQQKPGQPPKLLIY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="30">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q162">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2

LFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q163">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="31">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q164">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2 LFR4; anti-FR

LFR4</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q165">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>FGGGTKLEIKR</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="32">

<INSDSeq>

<INSDSeq_length>118</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..118</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q166">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2 VH</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..118</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q167">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLQESGPGLVKPSETLSLTCTVSGYSITSGYSWHWIRQPPGKGLEWM

GYIHYSDNTNYAPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARIYYGNSFVYWGQGTLVTVSS<

/INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="33">

<INSDSeq>

<INSDSeq_length>114</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..114</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q168">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2 VL</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..114</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q169">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQKPGQP

PKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYFYPYTFGGGTKLEIKR</INS

DSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="34">

<INSDSeq>

<INSDSeq_length>448</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..448</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q170">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2 HC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..448</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q171">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLQESGPGLVKPSETLSLTCTVSGYSITSGYSWHWIRQPPGKGLEWM

GYIHYSDNTNYAPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARIYYGNSFVYWGQGTLVTVSSA

STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV

PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE

VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL

PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL

DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="35">

<INSDSeq>

<INSDSeq_length>220</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..220</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q172">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-Claudin18.2 LC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..220</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q173">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQKPGQP

PKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYFYPYTFGGGTKLEIKRTVAAP

SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="36">

<INSDSeq>

<INSDSeq_length>10</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..10</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q174">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..10</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q175">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GYTFTGYFMN</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="37">

<INSDSeq>

<INSDSeq_length>17</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q176">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..17</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q177">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RIHPYDGDTFYNQKFQG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="38">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q178">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q179">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>YDGSRAMDY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="39">

<INSDSeq>

<INSDSeq_length>15</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q180">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LCDR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q181">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>KASQSVSFAGTSLMH</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="40">

<INSDSeq>

<INSDSeq_length>7</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q182">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LCDR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..7</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q183">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>RASNLEA</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="41">

<INSDSeq>

<INSDSeq_length>9</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q184">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LCDR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..9</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q185">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QQSREYPYT</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="42">

<INSDSeq>

<INSDSeq_length>25</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q186">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..25</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q187">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVVKPGASVKISCKAS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="43">

<INSDSeq>

<INSDSeq_length>14</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q188">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..14</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q189">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WVKQSPGQSLEWIG</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="44">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q190">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q191">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>KATLTVDKSSNTAHMELLSLTSEDFAVYYCTR</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="45">

<INSDSeq>

<INSDSeq_length>11</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q192">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HFR4</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..11</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q193">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WGQGTTVTVSS</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="46">

<INSDSeq>

<INSDSeq_length>23</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q194">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LFR1</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..23</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q195">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVLTQSPLSLAVSLGQPAIISC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="47">

<INSDSeq>

<INSDSeq_length>15</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q196">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LFR2</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..15</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q197">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>WYHQKPGQQPRLLIY</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="48">

<INSDSeq>

<INSDSeq_length>32</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q198">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LFR3</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..32</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q199">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>GVPDRFSGSGSKTDFTLTISPVEAEDAATYYC</INSDSeq_sequenc

e>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="49">

<INSDSeq>

<INSDSeq_length>118</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..118</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q200">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR VH</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..118</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q201">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIG

RIHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYDGSRAMDYWGQGTTVTVSS<

/INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="50">

<INSDSeq>

<INSDSeq_length>112</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..112</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q202">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR VL</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..112</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q203">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPR

LLIYRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPYTFGGGTKLEIKR</INSDS

eq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="51">

<INSDSeq>

<INSDSeq_length>448</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..448</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q204">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR HC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..448</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q205">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>QVQLVQSGAEVVKPGASVKISCKASGYTFTGYFMNWVKQSPGQSLEWIG

RIHPYDGDTFYNQKFQGKATLTVDKSSNTAHMELLSLTSEDFAVYYCTRYDGSRAMDYWGQGTTVTVSSA

STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV

PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE

VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL

PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL

DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

<SequenceData sequenceIDNumber="52">

<INSDSeq>

<INSDSeq_length>218</INSDSeq_length>

<INSDSeq_moltype>AA</INSDSeq_moltype>

<INSDSeq_division>PAT</INSDSeq_division>

<INSDSeq_feature-table>

<INSDFeature>

<INSDFeature_key>REGION</INSDFeature_key>

<INSDFeature_location>1..218</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier id="q206">

<INSDQualifier_name>note</INSDQualifier_name>

<INSDQualifier_value>anti-FR LC</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

<INSDFeature>

<INSDFeature_key>source</INSDFeature_key>

<INSDFeature_location>1..218</INSDFeature_location>

<INSDFeature_quals>

<INSDQualifier>

<INSDQualifier_name>mol_type</INSDQualifier_name>

<INSDQualifier_value>protein</INSDQualifier_value>

</INSDQualifier>

<INSDQualifier id="q207">

<INSDQualifier_name>organism</INSDQualifier_name>

<INSDQualifier_value>synthetic construct</INSDQualifier_value>

</INSDQualifier>

</INSDFeature_quals>

</INSDFeature>

</INSDSeq_feature-table>

<INSDSeq_sequence>DIVLTQSPLSLAVSLGQPAIISCKASQSVSFAGTSLMHWYHQKPGQQPR

LLIYRASNLEAGVPDRFSGSGSKTDFTLTISPVEAEDAATYYCQQSREYPYTFGGGTKLEIKRTVAAPSV

FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD

YEKHKVYACEVTHQGLSSPVTKSFNRGEC</INSDSeq_sequence>

</INSDSeq>

</SequenceData>

</ST26SequenceListing>

<---

Claims (15)

1. A drug-binding agent, wherein the drug-binding agent unit is selected from the following structures: 2. A conjugate comprising a binding agent-drug conjugate, wherein the binding agent-drug is an antibody-drug conjugate, wherein the antibody-drug conjugate is selected from the following structures: where Ab denotes an antibody or antigen-binding fragment thereof, where the antibody or antigen-binding fragment thereof targets a cell surface receptor or a tumor-associated antigen, and m denotes any number from 1 to 10. 3. The conjugate of claim 2, wherein the cell surface receptor or tumor-associated antigen comprises: HER2, HER3, claudin18.2, folate receptor α (FRα), BCMA, PSMA, TROP-2, CD19, CD20, CD22, CD30, CD79b, EGFR, c-Met and CEACAM5. 4. The conjugate of claim 2, wherein the antibody comprises a monoclonal antibody, a polyclonal antibody, a dimer, a polymer, a multispecific antibody, an intact antibody, an antibody fragment, a human antibody, a humanized antibody, or a chimeric antibody. 5. The conjugate of claim 2, wherein the antigen-binding fragment comprises Fab, Fab', Fv fragment, F(ab')2, scFv, di-scFv and/or dAb. 6. The conjugate of any one of claims 2-5, wherein the antibody is a monoclonal antibody. 7. The conjugate of any one of claims 2 to 6, wherein the antibody is a human antibody, a humanized antibody, or a chimeric antibody. 8. The conjugate of any one of claims 2-7, wherein the antibody or antigen-binding fragment thereof comprises: an anti-HER2 antibody, an anti-HER3 antibody, an anti-claudin18.2 antibody, an anti-FRα antibody, or fragments that bind its antigens. 9. A pharmaceutical composition for selective delivery of effective doses of cytotoxic agents or cytostatic agents to cancer cells or tumor tissues, comprising an effective amount of a conjugate according to any one of claims 2-8 and, optionally, a pharmaceutically acceptable carrier. 10. A method of treating a disease or condition in a patient in need thereof, comprising administering to the patient an effective amount of the drug binding agent of claim 1, the conjugate of any one of claims 2-8, or the pharmaceutical composition of claim 9; wherein the effective amount administered to the patient is typically from about 0.01 mg/kg to about 100 mg/kg of the patient's body weight or from 1.0 mcg/kg to 5.0 mg/kg of the patient's body weight; where the disease or condition includes a solid tumor and a hematological tumor.