RU2849370C1 - Methods and dosage regimens containing cdk2 inhibitor and cdk4 inhibitor for the treatment of cancer - Google Patents

Abstract

FIELD: combined cancer therapy.

SUBSTANCE: a group of inventions relates to combination cancer therapy. A method for treating cancer in a subject in need thereof comprises administering to the subject: (a) an amount of compound (A): (1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (PF-07104091) or its hydrate, (b) an amount of compound (B): 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropane-2-yl)-1-(propane-2-yl)-1H-benzimidazole-6-yl]pyrimidine-2- yl}amino)-2,3-dideoxy-D-threo-pentite (PF-07220060) or its hydrate, wherein the amounts of (a) and (b) together are effective in treating cancer, and wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, oesophageal cancer, liver cancer, pancreatic cancer, and stomach cancer. A combination for treating cancer is also disclosed.

EFFECT: higher efficacy than treatment with therapeutic agents alone.

19 cl, 6 tbl, 7 ex

Description

LEVEL OF TECHNOLOGY

Field of technology

The present invention relates to methods and combination therapies useful for treating cancer in a subject in need thereof. In particular, the invention relates to methods and combinations comprising a cyclin-dependent kinase 2 (CDK2) inhibitor such as (1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (hereinafter PF-07104091) or its monohydrate in combination with a selective cyclin-dependent kinase 4 (CDK4) inhibitor such as 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (hereinafter PF-07220060) or a pharmaceutically acceptable salt thereof, wherein the combination is optionally in additional combination with an additional anticancer agent, such as an endocrine therapy agent. The present invention also relates to corresponding dosage regimens, uses, and pharmaceutical compositions.

Description of the prior art

Cyclin-dependent kinases (CDKs) are important cellular enzymes that perform crucial functions in regulating eukaryotic cell division and proliferation. CDK inhibitors may be useful for the treatment of proliferative disorders, including cancer.

Overexpression of CDK2 is associated with abnormal cell cycle regulation. The cyclin E/CDK2 complex plays a critical role in regulating the G1/S transition, histone biosynthesis, and centrosome duplication. Progressive phosphorylation of retinoblastoma (RB) by cyclin D/Cdk4/6 and cyclin E/Cdk2 releases the G1 transcription factor, E2F, and promotes S-phase entry. Activation of cyclin A/CDK2 during early S-phase promotes phosphorylation of endogenous substrates that mediate DNA replication and inactivation of E2F for S-phase completion. (Asghar et al. The history and future of targeting cyclin-dependent kinases in cancer therapy, Nat. Rev. Drug. Discov. 2015; 14(2): 130–146).

Cyclin E, a regulatory cyclin of CDK2, is frequently overexpressed in cancer. Amplification or overexpression of cyclin E has long been associated with poor outcomes in breast cancer. (Keyomarsi et al., Cyclin E and survival in patients with breast cancer. N Engl J Med. (2002) 347:1566–75). Overexpression of cyclin E2 (CCNE2) is associated with endocrine resistance in breast cancer cells, and inhibition of CDK2 has been reported to restore sensitivity to tamoxifen or CDK4 inhibitors in tamoxifen-resistant, CCNE2-overexpressing cells. (Caldon et al., Cyclin E2 overexpression is associated with endocrine resistance but not insensitivity to CDK2 inhibition in human breast cancer cells. Mol. Cancer Ther. (2012) 11:1488–99; Herrera-Abreu et al., Early Adaptation and Acquired Resistance to CDK4/6 Inhibition in Estrogen Receptor-Positive Breast Cancer, Cancer Res. (2016) 76: 2301–2313). Cyclin E amplification has also been reported to contribute to trastuzumab resistance in human epidermal growth factor receptor 2 (HER2+)-positive breast cancer. (Scaltriti et al. Cyclin E amplification/overexpression is a mechanism of trastuzumab resistance in HER2+ breast cancer patients, Proc Natl Acad Sci. (2011) 108: 3761–6). Cyclin E overexpression has also been reported to play a role in basal-like and triple-negative breast cancer (TNBC), as well as inflammatory breast cancer. (Elsawaf & Sinn, Triple Negative Breast Cancer: Clinical and Histological Correlations, Breast Care (2011) 6:273–278; Alexander et al., Cyclin E overexpression as a biomarker for combination treatment strategies in inflammatory breast cancer, Oncotarget (2017) 8: 14897–14911.)

High CCNE1 mRNA expression was found to be associated with relative resistance to palbociclib, suggesting a role for CDK2 inhibition in reducing or overcoming resistance to CDK4/6 inhibition. (Turner et al., Cyclin E1 Expression and Palbociclib Efficacy in Previously Treated Hormone Receptor-Positive Metastatic Breast Cancer, J. Clin. Oncol. (2019) 37:1169–1178). Cyclin E1 (CCNE1) amplification or overexpression is also associated with poor outcomes in ovarian, gastric, endometrial, and other cancers. (Nakayama et al., Gene amplification CCNE1 is related to poor survival and potential therapeutic target in ovarian cancer, Cancer (2010) 116: 2621-34; Etemadmoghadam et al., Resistance to CDK2 Inhibitors Is Associated with Selection of Polyploid Cells in CCNE1-Amplified Ovarian Cancer, Clin Cancer Res (2013) 19: 5960-71; Au-Yeung et al., Selective Targeting of Cyclin E1-Amplified High-Grade Ovarian Cancer by Cyclin-Dependent Kinase 2 and ACT Inhibition, Clin. Cancer Res. (2017) 23:1862-1874; CCNE1 copy-number gain and overexpression identify ovarian clear cell carcinoma. poor prognosis, Modern Pathology (2017) 30: 297–303; Ooi et al., Gene amplification of CCNE1, CCND1, and CDK6 in gastric cancers detected by multiplex ligation-dependent probe amplification and fluorescence in situ hybridization, Hum Pathol. (2017) 61:58–67; Noske et al., Detection of CCNE1/URI (19q12) amplification by in situ hybridization is common in high grade and type II endometrial cancer, Oncotarget (2017) 8: 14794–14805).

CDK4 and CDK6 are important regulators of cell cycle progression at the G1-S checkpoint, which are controlled by D-type cyclins and INK4 endogenous CDK inhibitors such as p16 ^INK4a (CDKN2A). Dysregulation of the cyclin D-CDK4/6-INK4-retinoblastoma (Rb) pathway has been reported to be associated with the development of resistance to endocrine therapy.

CDK4/6 inhibition has emerged as a promising strategy for cancer therapy, particularly for the treatment of endocrine-resistant breast cancer (ERBC). (Rani, A., et. al., Endocrine Resistance in Hormone Receptor-Positive Breast Cancer—From Mechanism to Therapy. Front Endocrinol (Lausanne) 10:245, 2019).

CDK4/6 inhibitors (eg, palbociclib, abemaciclib, ribociclib), when dosed in combination with endocrine therapy, significantly improve progression-free survival and/or overall survival in patients with HR-positive/HER2-negative advanced or metastatic breast cancer. (Spring, L.M., et. al., Cyclin-dependent kinase 4 and 6 inhibitors for hormone receptor-positive breast cancer: past, present, and future. Lancet, 395, 817-827, 2020). However, CDK4/6 inhibitors are associated with dose-limiting hematologic toxicity, primarily neutropenia, and gastrointestinal toxicity. As with other kinase inhibitors, the efficacy of CDK4/6 inhibitors may be limited over time by the development of primary or acquired resistance.

New data suggest that cyclin D3-CDK6 may be associated with the observed hematological toxicity. (Malumbres et al., Mammalian Cells Cycle without the D-type Cyclin-Dependent Kinases Cdk4 and Cdk6, (2004) Cell 118(4): 493–504; Sicinska et al. Essential Role for Cyclin D3 in Granulocyte Colony-Stimulating Factor-Driven Expansion of Neutrophil Granulocytes (2006), Mol. Cell Biol 26(21): 8052–8060; Cooper et al. A unique function for cyclin D3 in early B cell development, (2006), Nat. Immunol. 5(7): 489–497). CDK4 has been identified as the sole oncogenic factor in many breast cancers. Accordingly, a selective CDK4 inhibitor may provide an improved safety profile or increased overall efficacy due to the possibility of higher and/or continuous dosing compared to dual CDK4/6 inhibitors.

The compound (1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (hereinafter PF-07104091) is a potent and selective CDK2 inhibitor with the structure:

PF-07104091 (Pfizer Inc.) is currently in clinical development for the treatment of certain types of cancer. The preparation of PF-07104091 is disclosed in International Patent Publication No. WO 2020/157652 and U.S. Patent No. 11,014,911, the contents of each of which are incorporated herein by reference in their entirety. PF-07104091 also has the name (1R,3S)-3-(3-(3-(methoxymethyl)-1-methyl-1H-pyrazole-5-carboxamido)-1H-pyrazol-5-yl)cyclopentylisopropylcarbamate, generated in ChemDraw 20.1.1.

The compound 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (hereinafter PF-07220060) is a potent and selective CDK4 inhibitor with the structure:

or its pharmaceutically acceptable salt.

PF-07220060 (Pfizer Inc.) is currently in clinical development for the treatment of certain types of cancer. The preparation of PF-07220060 is disclosed in International Patent Publication No. WO 2019/207463 and U.S. Patent No. 10,766,884, the contents of each of which are incorporated herein by reference in their entirety. PF-07220060 also has the name (3S,4R)-4-((5-chloro-4-(4-fluoro-2-(2-hydroxypropan-2-yl)-1-isopropyl-1H-benzo[d])imidazol-6-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3-ol, generated in ChemDraw 20.1.1.

There remains a need for improved therapies for the treatment of cancer. The combinations, methods, dosing regimens, and administration disclosed herein are believed to have one or more advantages, such as greater efficacy than treatment with any single therapeutic agent; the potential to reduce drug-drug interactions; the potential to improve dosing regimens; the potential to reduce side effects; the potential to overcome resistance mechanisms, and the like.

THE DRYNESS OF THE INVENTION

The present invention relates to methods, combinations, dosage regimens, uses and pharmaceutical compositions for the treatment of cancer in a subject in need thereof, comprising a selective CDK2 inhibitor of Formula (I) or a pharmaceutically acceptable salt or solvate thereof, and a selective CDK4 inhibitor, such as a compound of Formula (II) or a pharmaceutically acceptable salt thereof.

In one aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject: (a) an amount of a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) the amount of a selective inhibitor of cyclin-dependent kinase 4 (CDK4);

where quantities (a) and (b) together are effective in treating cancer.

In some embodiments of this aspect, the invention provides a method further comprising administering to the subject: (c) an amount of an additional anti-cancer agent; wherein the amounts (a), (b), and (c) are together effective in treating cancer.

In another aspect, the invention provides a combination comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) a selective inhibitor of cyclin-dependent kinase 4 (CDK4);

where the combination of (a) and (b) is effective in treating cancer.

In some embodiments of this aspect, the combination further comprises (c) an additional anti-cancer agent; wherein the combination of (a), (b) and (c) is effective in treating cancer.

In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ⁵ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) selective inhibitor of cyclin-dependent kinase 4 (CDK4).

In some embodiments of this aspect, the combination for use further comprises (c) an additional anticancer agent.

In another aspect, the invention provides the use of a combination comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) a selective inhibitor of cyclin-dependent kinase 4 (CDK4);

where the use of the combination is effective in the treatment of cancer.

In some embodiments of this aspect, the combination further comprises (c) an additional anti-cancer agent, wherein use of the combination of (a), (b) and (c) is effective in treating cancer.

In preferred embodiments of each of the methods, combinations, and uses herein, the compound of Formula (I) is {1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (PF-07104091), a potent CDK2 inhibitor having the structure:

or its monohydrate.

In particularly preferred embodiments of each of the methods, combinations and uses described herein, the compound of Formula (I) is PF-07104091 monohydrate.

In some embodiments of each of the methods, combinations, and uses described herein, the combination of a compound of Formula (I) and a selective CDK4 inhibitor is synergistic. In some embodiments of the methods, combinations, and uses described herein, the combination of a compound of Formula (I), a CDK4 inhibitor, and an additional anticancer agent is synergistic.

In some embodiments of each of the methods, combinations, and uses described herein, the selective CDK4 inhibitor is a compound of Formula (II):

or a pharmaceutically acceptable salt thereof, where:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ independently represents H or C ₁ -C ₂ alkyl.

In preferred embodiments, the compound of Formula (II) is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threopentitol (PF-07220060), a potent and selective CDK4 inhibitor, having the structure:

or a pharmaceutically acceptable salt thereof.

In preferred embodiments of each of the methods, combinations, and uses described herein, the compound of Formula (I) is PF-07104091 or a pharmaceutically acceptable solvate thereof, and the compound of Formula (II) is PF-07220060 or a pharmaceutically acceptable salt thereof. In particularly preferred embodiments of each of the methods, combinations, and uses described herein, the compound of Formula (I) is PF-07104091 monohydrate, and the compound of Formula (II) is PF-07220060.

In some embodiments of each of the methods, combinations, and uses described herein, the invention further includes one or more additional anti-cancer agents. In preferred embodiments, the cancer is breast cancer (including HR+/HER2- breast cancer), and the additional anti-cancer agent is an endocrine therapeutic agent. In some such embodiments, the endocrine therapeutic agent is an aromatase inhibitor, a selective estrogen receptor degrader (SERD), or a selective estrogen receptor modulator (SERM). In preferred embodiments, the endocrine therapeutic agent is letrozole or fulvestrant.

Embodiments of each of the methods, combinations and applications described herein may be combined with one or more other embodiments, provided that such embodiments do not contradict each other.

DETAILED DESCRIPTION

The present disclosure can be more readily understood by reference to the following detailed description of preferred embodiments of the invention and the Examples included therein. It should be understood that the terminology used herein is intended to describe particular embodiments only and is not intended to be limiting. Furthermore, it should be understood that, unless otherwise specifically indicated herein, the terminology used herein is to be given its conventional meaning as known in the relevant art.

E1. A method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) the amount of a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) the amount of compound of Formula (II):

or its pharmaceutically acceptable salt, where:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyl and C ₁ -C ₄ fluoroalkyl is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ is independently H or C ₁ -C ₄ alkyl;

where quantities (a) and (b) together are effective in treating cancer.

E2. The method of embodiment E1, further comprising administering to the subject: (c) an amount of an additional anti-cancer agent; wherein the amounts (a), (b), and (c) together are effective in treating cancer.

E3. The method according to embodiment E1 or E2, wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, and stomach cancer.

E4. The method of embodiment E2 or E3, wherein the cancer is hormone receptor positive (HR+) breast cancer and the additional anticancer agent is an endocrine therapeutic agent selected from the group consisting of an aromatase inhibitor, a selective estrogen receptor modulator (SERM), and a selective estrogen receptor biodegrader (SERD).

E5. The method of embodiment E4, wherein the endocrine therapeutic agent is letrozole or fulvestrant.

E6. The method according to any one of embodiments E1-E3, wherein the cancer is lung cancer.

E7. The method of embodiment E6, wherein the lung cancer is small cell lung cancer (SCLC).

E8. The method of embodiment E7, wherein the SCLC is characterized by loss of retinoblastoma (RB) function.

E9. The method of embodiment E6, wherein the lung cancer is non-small cell lung cancer (NSCLC).

E10. The method of embodiment E9, wherein the NSCLC is lung squamous cell carcinoma (LUSC) or lung adenocarcinoma (LUAD).

E11. The method of embodiment E10, wherein the NSCLC is KRAS-associated lung adenocarcinoma (LUAD).

E12. The method according to any one of embodiments E1-E11, wherein the compound of Formula (I) is (1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate monohydrate (PF-07104091).

E13. The method of embodiment E12, wherein the amount of PF-07104091 is from about 75 mg to about 500 mg BID.

E14. The method of embodiment E13, wherein the amount of PF-07104091 is about 75 mg BID, about 150 mg BID, or about 225 mg BID.

E15. The method of embodiment E12, wherein the amount of PF-07104091 is from about 100 mg to about 500 mg BID.

E16. The method of embodiment E15, wherein the amount of PF-07104091 is from about 150 mg to about 300 mg BID.

E17. The method of embodiment E15 or E16, wherein the amount of PF-07104091 is about 150 mg BID, about 225 mg BID, or about 300 mg BID.

E18. The method according to any one of embodiments E1-E17, wherein the compound of Formula (II) is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (PF-07220060) or a pharmaceutically acceptable salt thereof.

E19. The method of embodiment E18, wherein the amount of PF-07220060 is from about 100 mg to about 500 mg BID.

E20. The method of embodiment E19, wherein the amount of PF-07220060 is from about 300 mg to about 500 mg BID.

E21. The method of embodiment E20, wherein the amount of PF-07220060 is about 100 mg BID, about 200 mg BID, or about 300 mg BID.

E22. The method of embodiment E19 or E20, wherein the amount of PF-07220060 is about 300 mg BID or about 400 mg BID.

E23. Combination containing:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) a compound of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein: R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ is independently H or C ₁ -C ₄ alkyl;

where the combination of (a) and (b) is effective in treating cancer.

E24. The combination according to embodiment E23, further comprising (c) an additional anticancer agent; wherein the combination of (a), (b) and (c) is effective in treating cancer.

E25. The combination according to embodiment E23 or E24, wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer and stomach cancer.

E26. The combination of embodiment E24 or E25, wherein the cancer is hormone receptor positive (HR+) breast cancer and the additional anticancer agent is an endocrine therapeutic agent selected from the group consisting of an aromatase inhibitor, a selective estrogen receptor modulator (SERM), and a selective estrogen receptor biodegrader (SERD).

E27. The combination of embodiment E26, wherein the endocrine therapeutic agent is letrozole or fulvestrant.

E28. The combination according to any one of embodiments E23-E25, wherein the cancer is lung cancer.

E29. The combination of embodiment E28, wherein the lung cancer is small cell lung cancer (SCLC).

E30. The combination of embodiment E29, wherein the SCLC is characterized by loss of function of retinoblastoma (RB).

E31. The combination of embodiment E28, wherein the lung cancer is non-small cell lung cancer (NSCLC).

E32. The combination of embodiment E31, wherein the NSCLC is lung squamous cell carcinoma (LUSC) or lung adenocarcinoma (LUAD).

E33. The combination of embodiment E32, wherein the NSCLC is KRAS-associated lung adenocarcinoma (LUAD).

E34. The combination according to any one of embodiments E23-E33, wherein the compound of Formula (I) is (1R,35)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (PF-07104091) monohydrate.

E35. The combination of embodiment E34, wherein the amount of PF-07104091 is from about 50 mg to about 250 mg BID.

E36. The combination of embodiment E35, wherein the amount of PF-07104091 is from about 75 mg to about 150 mg BID.

E37. A combination of embodiments E35 or E36, wherein the amount of PF-07104091 is about 75 mg BID, about 100 mg BID, about 125 mg BID, or about 150 mg BID.

E38. The combination of embodiment E35, wherein the amount of PF-07104091 is about 75 mg BID, about 150 mg BID, or about 225 mg BID.

E39. A combination of any one of embodiments E23-E38, wherein the compound of Formula (II) is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (PF-07220060) or a pharmaceutically acceptable salt thereof.

E40. The combination of embodiment E39, wherein the amount of PF-07220060 is from about 100 mg to about 500 mg BID.

E41. The combination of embodiment E40, wherein the amount of PF-07220060 is from about 300 mg to about 500 mg BID.

E42. The combination according to embodiment E40 or E41, wherein the amount of PF-07220060 is about 300 mg BID or about 400 mg BID.

E43. The method of embodiment E40, wherein the amount of PF-07220060 is about 100 mg BID, about 200 mg BID, or about 300 mg BID.

E44. A method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities PF-07104091; and

(b) an amount of PF-07220060 or a pharmaceutically acceptable salt thereof;

where quantities (a) and (b) together are effective in treating cancer.

E45. A method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities PF-07104091;

(b) an amount of PF-07220060, or a pharmaceutically acceptable salt thereof; and

(c) the amount of additional anticancer agent;

where quantities (a), (b) and (c) together are effective in treating cancer.

E46. The method according to embodiment E44 or E45, wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, and stomach cancer.

E47. The method according to any one of embodiments E44-E46, wherein the cancer is hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-) breast cancer.

E48. The method according to any one of embodiments E45-E47, wherein the additional anticancer agent is an endocrine therapeutic agent selected from the group consisting of an aromatase inhibitor, a SERM, and a SERD.

E49. The method according to embodiment E48, wherein the endocrine therapeutic agent is letrozole or fulvestrant.

E50. The method according to any one of embodiments E44-E46, wherein the cancer is lung cancer.

E51. The method according to embodiment E50, wherein the lung cancer is small cell lung cancer (SCLC).

E52. The method of embodiment E51, wherein the SCLC is characterized by loss of retinoblastoma (RB) function.

E53. The method according to embodiment E50, wherein the lung cancer is non-small cell lung cancer (NSCLC).

E54. The method of embodiment E53, wherein the NSCLC is lung squamous cell carcinoma (LUSC) or lung adenocarcinoma (LUAD).

E55. The method of embodiment E54, wherein the NSCLC is KRAS-associated lung adenocarcinoma (LUAD).

E56. The method according to any one of embodiments E44-E55, wherein the amount of PF-07104091 is from about 50 mg to about 250 mg BID.

E57. The method of embodiment E56, wherein the amount of PF-07104091 is from about 75 mg to about 150 mg BID.

E58. The method of embodiment E56 or E57, wherein the amount of PF-07104091 is about 75 mg BID, about 100 mg BID, about 125 mg BID, or about 150 mg BID.

E59. The method of embodiment E56 or E57, wherein the amount of PF-07104091 is about 75 mg BID, about 150 mg BID, or about 225 mg BID.

E60. The method according to any one of embodiments E44-E59, wherein the compound of Formula (II) is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (PF-07220060) or a pharmaceutically acceptable salt thereof.

E61. The method of embodiment E60, wherein the amount of PF-07220060 is from about 100 mg to about 500 mg BID.

E62. The method of embodiment E61, wherein the amount of PF-07220060 is from about 300 mg to about 500 mg BID.

E63. The method of embodiment E61, wherein the amount of PF-07220060 is about 100 mg BID, about 200 mg BID, or about 300 mg BID.

E64. The method according to embodiment E61 or E62, wherein the amount of PF-07220060 is about 300 mg BID or about 400 mg BID.

E65. The method according to any one of embodiments E1-E22, wherein the cancer is a cancer resistant to a CDK4/6 inhibitor.

E66. The method of embodiment E65, wherein the CDK4/6 inhibitor is palbociclib.

E67. The method according to any one of embodiments E1-E22, wherein the subject has previously been treated with a CDK4/6 inhibitor.

E68. The method of embodiment E67, wherein the CDK4/6 inhibitor is palbociclib.

Definitions:

As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise noted. For example, the substituent "a" includes one or more substituents.

The disclosure described herein may be suitably practiced in the absence of any element(s) not specifically disclosed herein. Thus, for example, in each instance herein, any of the terms "comprising," "consisting essentially of," and "consisting of" may be replaced by either of the other two terms.

The term "about" means a value within the accepted standard error of the mean, as considered by one of ordinary skill in the art. In some embodiments, the term "about" means within ±10% of the stated value. For example, a dose of approximately 150 mg should be understood to mean that the dose can range from 135 mg to 165 mg.

The terms "cancer," "cancerous," or "malignant" refer to or describe a physiological condition in mammals typically characterized by unregulated cell growth. As used herein, cancer can refer to any malignant and/or invasive growth or tumor caused by abnormal cell growth, including solid tumors, named for the cell type that forms them, as well as cancers of the blood, bone marrow, or lymphatic system. Examples of solid tumors include, but are not limited to, sarcomas and carcinomas. Examples of blood cancers include, but are not limited to, leukemia, lymphoma, and myeloma. The term "cancer" includes, but is not limited to, a primary cancer that occurs in a specific area of the body, metastatic cancer that has spread from the site where it began to other parts of the body, a recurrence of the original primary cancer after remission, or a second primary cancer, which is a new primary cancer in a person who has had a previous cancer of a type different from the last.

As used herein, the term "PF-07104091" may refer to the free base of PF-07104091 and/or the monohydrate of PF-07104091. In preferred embodiments, PF-07104091 as referred to herein is the monohydrate of PF-07104091.

As used herein, the term "dose-limiting toxicity" (DLT) refers to a dose of, for example, PF-07104091 or another agent described herein, that is a contraindication for further dosage escalation.

As used herein, the "maximum tolerated dose" (MTD) refers to the highest dose of PF-07104091 or another agent described herein that does not cause unacceptable side effects or intolerable toxicity. The MTD is estimated using the mTPI based on the observed degree of DLT.

The terms "patient" or "subject" refer to any individual subject for whom therapy is desired or who participates in a clinical trial, epidemiological study, or is used as a control, including humans and mammalian veterinary patients, such as domestic animals such as cattle, horses, dogs, and cats; non-human primates such as monkeys; laboratory animals such as rats, mice, and guinea pigs; and captive wild animals such as lions, tigers, and the like. In preferred embodiments, the subject is a human.

In some embodiments, the subject is an adult. In some embodiments, the adult subject is a woman in any state of menopause or a man. In some such embodiments, the subject is a postmenopausal woman or a man. In some such embodiments, the subject is a postmenopausal woman. In some such embodiments, the subject is a premenopausal or perimenopausal woman. In some such embodiments, the subject is a premenopausal or perimenopausal woman receiving a luteinizing hormone-releasing hormone (LHRH) agonist. In some such embodiments, the subject is a man. In some such embodiments, the subject is a man receiving an LHRH agonist. In some embodiments, the subject is a child between the ages of birth and 18 years. In some embodiments, the subject is a child between the ages of birth and 15 years suffering from pediatric cancer.

For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: reducing the proliferation (or killing) of neoplastic or cancer cells; inhibiting metastasis or neoplastic cells; shrinking or reducing the size of a tumor; remission of cancer; reducing symptoms resulting from cancer; improving the quality of life of cancer patients; reducing the dose of other drugs needed to treat cancer; delaying the progression of cancer; curing cancer; overcoming one or more mechanisms of cancer resistance; and/or prolonging the survival of patients with cancer. Positive therapeutic effects in cancer can be measured in several ways (see, e.g., W.A. Weber, Assessing tumor response to therapy, J. Nucl. Med. 50 Suppl. 1:1S-10S (2009). For example, with respect to tumor growth inhibition (TGI), according to National Cancer Institute (NCI) standards, a TGI of less than or equal to 42% represents a minimal level of antitumor activity; a TGI of <10% is considered a high level of antitumor activity, where TGI (%) = average treated tumor volume / average control tumor volume × 100.

The therapeutic effect of the methods, combinations, and uses described herein can be determined with reference to any of the following: partial response (PR), complete response (CR), progression-free survival (PFS), disease-free survival (DFS), duration of response (DoR), overall response rate (ORR), or overall survival (OS). In some embodiments, the response to the combination, method, or use described herein is any of PR, CR, PFS, DFS, DoR, ORR, or OS, as assessed using the RECIST v1.1 response criteria (Eisenhauer et al., New response evaluation criteria in solid tumors: Revised RECIST guideline (version 1.1), Eur J of Cancer, 2009; 45(2):228-47).

In some embodiments, the methods, combinations, and uses herein relate to neoadjuvant therapy, adjuvant therapy, first-line therapy, second-line therapy, or third-line or subsequent therapy. In each case, as further described herein, the cancer may be localized, advanced, or metastatic, and the intervention may occur at any point along the disease continuum (i.e., at any stage of the cancer).

A treatment regimen for a combination, method, or application that is effective for treating cancer in a subject may vary depending on factors such as the disease state, age, and weight of the subject, and the ability of the therapy to elicit an anti-cancer response in the subject. Although an embodiment of any of the aspects disclosed herein may not be effective in achieving a positive therapeutic effect in every subject, it should have such an effect on a statistically significant number of subjects, as determined by any statistical test known in the art, such as the Student t-test, the chi-square test, the Mann-Whitney U-test, the Kruskal-Wallis (H-test), the Jonckheere-Terpstraat test, and the Wilcon test.

As used herein, an "effective amount" or "therapeutically effective amount" for use in and/or treatment of a subject refers to an amount that provides, in a single or multiple doses, alone or in combination with one or more other agents, treatments, protocols, or therapeutic regimens, a detectable response of any duration (temporary, medium-term, or long-term), a desired outcome, or an objective or subjective benefit to the subject to any measurable or detectable degree or over any period of time (e.g., hours, days, months, years, remission, or cure). Such amounts are generally effective in alleviating a disease or one, some, or all of the side effects/symptoms, sequelae, or complications of the disease to a measurable degree, although a reduction or suppression of the progression or worsening of the disease or ensuring stability (i.e., no worsening) of the disease condition is considered a satisfactory outcome. The term "therapeutically effective amount" also means an amount of an active agent effective to produce a desired therapeutic effect when administered to a subject, such as stopping the growth or shrinking a cancer tumor.

The effective amount may vary depending on factors such as the disease state, age, gender, and weight of the subject. For prophylactic use, beneficial or desired outcomes may include eliminating or reducing the risk, reducing the severity, or delaying the onset of the disease. For therapeutic use, beneficial or desired outcomes may include reducing the incidence or alleviating one or more symptoms of the disease, reducing the dose of another drug used to treat the disease, improving the efficacy or safety of another drug used to treat the disease, delaying the time to disease progression, or prolonging survival.

As used herein, a "treatment cycle" refers to a period of time involving the administration of one or more agents, with or without rest periods between treatment cycles. A treatment cycle may be continuous, i.e., without a rest period between treatment cycles. Alternatively, a treatment cycle may be intermittent and include a rest period (i.e., a period of discontinued dosing of one or more days or weeks without treatment) between treatment cycles. In such cases, administration of another agent during the rest period should not interfere with or compromise the administration of the agent(s) described herein.

For example, a 21-day or 28-day treatment cycle with 14 or 21 days of treatment, respectively, followed by a 7-day rest period (i.e., a treatment break) is an example of an intermittent treatment cycle. Treatment cycles consisting of 2 or 3 weeks of treatment and 1 week of treatment break are sometimes referred to as 2/1-week or 3/1-week treatment cycles, respectively. Alternatively, an intermittent treatment cycle may include a 7-day cycle with 5 days of treatment and 2 days of treatment break.

As used herein, the term "improvement" refers to any reduction in the degree, severity, frequency, and/or likelihood of occurrence of a symptom or clinical sign characteristic of a specific disease. "Symptom" refers to any subjective evidence of a disease or condition in a subject.

As used herein, the term "treating" or "treating" cancer and/or a cancer-associated disease means administering a mono- or combination therapy according to the present invention to a subject, patient, or individual having or diagnosed with cancer to achieve at least one beneficial therapeutic effect, such as, for example, reducing the number of cancer cells, reducing the size of a tumor, reducing the rate of infiltration of cancer cells into peripheral organs, or reducing the rate of tumor metastasis or tumor growth, reversing, alleviating, inhibiting the progression of, or preventing the disorder or condition to which the term applies, or one or more symptoms of such disorder or condition. The term "treating," as used herein, unless otherwise specified, refers to the act of treating as "treating" is defined immediately above. The term "treating" also includes adjuvant and neoadjuvant treatment of a subject. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, one or more of the following: reducing the proliferation (or destruction) of neoplastic or cancer cells; inhibiting metastasis or neoplastic cells; shrinking or decreasing the size of a tumor; remission of cancer; reduction in symptoms resulting from cancer; improving the quality of life of cancer patients; reducing the dose of other drugs needed to treat cancer; delaying the progression of cancer; curing cancer; overcoming one or more mechanisms of cancer resistance; and/or prolonging the survival of patients with cancer. Positive therapeutic effects in cancer can be measured in a variety of ways (see, e.g., W.A. Weber, J. Nucl. Med. 50:1S-10S (2009)).

The term "tumor," when used to describe a subject diagnosed or suspected of having cancer, refers to a malignant or potentially malignant neoplasm or tissue mass of any size and includes primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue that typically does not contain cysts or fluid-filled areas. Examples of solid tumors include sarcomas, carcinomas, and lymphomas. Leukemias (blood cancers) do not typically form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

As used herein, the terms "combination" or "combination therapy" refer to the administration of two or more therapeutic agents in a combination therapy, either as compounds or as a pharmaceutical composition or medicinal product. Combination therapy may be administered sequentially, consecutively, or simultaneously.

When administering combination therapy of two or more agents, the agents can be administered in the same treatment cycle or in different cycles. In preferred embodiments, PF-07104091 is administered continuously over a 28-day cycle. In preferred embodiments, PF-07220060 is administered continuously over a 28-day cycle. Fulvestrant is typically administered intramuscularly on days 1, 15, and 29 of the first treatment cycle and once monthly thereafter. Letrozole is typically administered continuously over a 28-day treatment cycle.

Each therapeutic agent of the methods and combination therapies described herein can be administered as a compound or as a pharmaceutical composition (also referred to herein as a drug) that comprises the therapeutic agent and one or more pharmaceutically acceptable carriers, excipients, or diluents in accordance with pharmaceutical practice.

The term "sequential" or "sequentially" refers to the administration of each therapeutic agent of a combination therapy, either individually or as part of a medicinal product, one after the other, where each therapeutic agent can be administered in any order. Sequential administration can be particularly useful when the therapeutic agents in the combination therapy are in different dosage forms, such as one agent is a tablet and the other is a sterile liquid, and/or when the agents are administered according to different dosing schedules, such as one agent administered daily and the second agent administered less frequently, such as weekly.

The term "sequential" refers to the administration of each therapeutic agent in a combination therapy, either separately or as separate drugs, where the second therapeutic agent is administered immediately after the first therapeutic agent, but these therapeutic agents can be administered in any order. In a preferred embodiment, the therapeutic agents are administered sequentially.

The term "concomitant" refers to the administration of each therapeutic agent of a combination therapy in the same dosage form, such as a fixed-dose combination comprising two or more drugs in a single dosage form.

A "dosage regimen" refers to a period of administration of one or more medicinal products, compounds, or compositions, comprising one or more treatment cycles, where each treatment cycle may include administration of one or more agents at different times, with different frequencies, or in different amounts, using the same or different routes of administration. Repeated administrations or dosing regimens or adjustments to the administration or dosing regimen may be made as needed to achieve the desired treatment effect.

"BID" or "bid" refers to the administration of a drug, compound, or composition twice daily.

"QD" or "qd" refers to once-daily administration of a drug, compound, or composition.

"TID" or "tid" refers to the administration of a drug, compound, or composition three times a day.

The term "additive" means that the result of a combination of two or more drugs, compounds, or compositions is not greater than the sum of the effects of each drug, compound, or composition individually.

The term "synergy" or "synergistic" means that the effect of a combination of two or more drugs, compounds, or compositions is greater than the sum of the effects of each drug, compound, or composition individually. Combinations that provide a synergistic effect are called synergistic combinations.

"Synergistic amount" is the amount of two or more drugs, compounds, or compositions in combination that results in a synergistic effect. The synergistic effect can be calculated, for example, using suitable methods such as the Sigmoid-Emax equation (Holford, N.H.G. and Scheiner, L.B., Clin. Pharmacokinet. 6: 429-453 (1981)), the Loewe additivity equation (Loewe, S. and Muischnek, H., Arch. Exp.Pathol. Pharmacol. 114: 313-326 (1926)) and the median effect equation (Chou, T.C. and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each equation mentioned above can be applied to experimental data to generate an appropriate graph to help evaluate the effects of the drug combination.

The corresponding graphs associated with the above equations are the concentration-effect curve, the isobologram curve, and the combined index curve, respectively. Ma & Motsinger-Reif, Current Method for Quantifying Drug Synergism, Proteom. Bioinform (2019) 1 (2):43–48; Tang et al., What is Synergy? The Saariselka Agreement Revisited, Front Pharmacol. (2015) Article 181.6: 1–5.

All references herein to CDK2 inhibitors of Formula (I) or to CDK4 inhibitors of Formula (II) include (to the extent chemically feasible) references to their pharmaceutically acceptable salts, solvates, hydrates and complexes, and to solvates, hydrates and complexes of their pharmaceutically acceptable salts, and include amorphous and polymorphic forms, stereoisomers and isotopically labeled versions thereof.

As used herein, the term "pharmaceutically acceptable salt" refers to those salts that retain the biological effectiveness and properties of the parent compound. The phrase "pharmaceutically acceptable salt(s)" as used herein, unless otherwise specified, includes salts of acidic or basic groups that may be present in the compounds of the formulae disclosed herein. For example, compounds having a basic nature can form a variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are selected from those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions. Examples of anions suitable for mono- and di-acid addition salts include, but are not limited to, acetate, aspartate, benzenesulfonate, benzoate, besylate, bicarbonate, bisulfate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, decanoate, edetate, edislyate, estalate, esylate, fumarate, gluceptate, gluconate, glutamate, glycolate, hexanoate, hexylresorcinate, hydrabamine, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methyl sulfate, mucate, napsylate, nitrate, octanoate, oleate, pamoate (embonate), pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, theoclate, tosylate, triethiodide, and valerate. Alternatively, compounds that are acidic in nature may be capable of forming base salts with various pharmacologically acceptable cations that form non-toxic base salts. Such non-toxic base salts include, but are not limited to, salts derived from such pharmacologically acceptable cations as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium, or water-soluble amine addition salts such as N-methylglucamine (meglumine), and lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines. Examples of cations suitable for such salts include alkali or alkaline earth metal salts and other cations, including aluminum, arginine, benzathine, calcium, chloroprocaine, choline, diethanolamine, ethanolamine, ethylenediamine, lysine, magnesium, histidine, lithium, meglumine, potassium, procaine, sodium, triethiamine, and zinc. The salts can be prepared by conventional methods. Hemisalts of acids and bases, such as hemisulfate and hemicalcium salts, can also be formed. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts are known to those skilled in the art. Therapeutic methods, combinations, uses The present invention provides methods, combinations and uses comprising a CDK2 inhibitor which is a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ⁵ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy.

In each instance provided herein, reference to “a compound of Formula (I)” may be replaced by “a CDK2 inhibitor of Formula (I)”.

The present invention further provides methods, combinations and uses comprising a selective CDK4 inhibitor that is a compound of Formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ independently represents H or C ₁ -C ₂ alkyl.

In one aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject: (a) an amount of a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) the amount of compound of Formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ independently represents H or C ₁ -C ₂ alkyl;

where quantities (a) and (b) together are effective in treating cancer.

In some embodiments, the method further comprises administering to the subject (c) an amount of an additional anti-cancer agent; wherein the amounts (a), (b), and (c) are together effective in treating cancer.

In another aspect, the invention provides a combination comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and (b) a compound of Formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ is independently H or C ₁ -C ₂ alkyl;

where the combination of (a) and (b) is effective in treating cancer.

In some embodiments, the combination further comprises (c) an additional anti-cancer agent; wherein the combination of (a), (b) and (c) is effective in treating cancer.

In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) a compound of Formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl, or C ₁ -C ₄ fluoroalkyl, where

each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ independently represents H or C ₁ -C ₂ alkyl.

In some embodiments, the combination for use further comprises (c) an additional anticancer agent.

In another aspect, the invention provides the use of a combination comprising:

(a) a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C ₄ alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy; and

(b) a compound of Formula (II):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ⁴ is H, F or Cl;

R ⁵ is H, C ₁ -C ₅ alkyl, C ₁ -C ₅ fluoroalkyl or C ₃ -C ₈ cycloalkyl, wherein each of said C ₁ -C ₅ alkyl and C ₁ -C ₅ fluoroalkyl is optionally substituted with R ⁸ and each of said C ₃ -C ₈ cycloalkyl is optionally substituted with R ⁹ ;

R ⁶ is H, C ₁ -C ₄ alkyl or C ₁ -C ₄ fluoroalkyl, wherein each of said C ₁ -C ₄ alkyls and C ₁ -C ₄ fluoroalkyls is optionally substituted with R ⁸ ;

R ⁷ is H, F or Cl;

each R ⁸ is independently OH, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ;

each R ⁹ is independently F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or NR ¹⁰ R ¹¹ ; and

each R ¹⁰ and R ¹¹ independently represents H or C ₁ -C ₂ alkyl;

where the use of a combination of (a) and (b) is effective in the treatment of cancer.

In some embodiments of this aspect, the combination further comprises (c) an additional anti-cancer agent, wherein use of the combination of (a), (b) and (c) is effective in treating cancer.

In some embodiments of each of the methods, combinations, and uses described herein, the compound of Formula (I) is (1R,3S)-3-[3-({[3-(methoxymethyl)-1-methyl-1H-pyrazol-5-yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentylpropan-2-ylcarbamate (PF-07104091), having the structure:

It is understood that the above structural formula of PF-07104091 includes all tautomeric forms that can coexist and directly interconvert under appropriate conditions. For example, in some embodiments, PF-07104091 has the structure:

In preferred embodiments of each of the described methods, combinations and uses described herein, the compound of Formula (II) is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threopentitol (PF-07220060), having the structure:

or its pharmaceutically acceptable salt.

In another aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities of PF-07104091 monohydrate; and

(b) an amount of PF-07220060 or a pharmaceutically acceptable salt thereof;

where quantities (a) and (b) together are effective in treating cancer.

In another aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities of PF-07104091 monohydrate;

(b) the amount of PF-07220060 or a pharmaceutically acceptable salt thereof; and

(c) the amount of additional anticancer agent;

where quantities (a), (b) and (c) together are effective in treating cancer.

In another aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities of PF-07104091 monohydrate; and

(b) the amount of selective CDK4 inhibitor;

where quantities (a) and (i) are together effective in treating cancer.

In another aspect, the invention provides a method of treating cancer in a subject in need thereof, comprising administering to the subject:

(a) quantities of PF-07104091 monohydrate;

(b) the amount of a selective CDK4 inhibitor; and

(c) the amount of additional anticancer agent;

where quantities (a), (b) and (c) together are effective in treating cancer.

In another aspect, the invention provides a combination comprising:

(a) PF-07104091 monohydrate; and

(b) PF-07220060 or a pharmaceutically acceptable salt thereof; wherein the combination of (a) and (b) is effective in treating cancer.

In another aspect, the invention provides a combination comprising:

(a) monohydrate PF-07104091;

(b) PF-07220060 or a pharmaceutically acceptable salt thereof; and an additional anticancer agent;

wherein the combination of (a), (b) and (c) is effective in the treatment of cancer. In another aspect, the invention provides a combination comprising:

(a) PF-07104091 monohydrate; and

(b) selective CDK4 inhibitor;

wherein the combination of (a) and (b) is effective in the treatment of cancer. In another aspect, the invention provides a combination comprising:

(a) monohydrate PF-07104091;

(b) a selective CDK4 inhibitor; and

(c) additional anticancer agent;

wherein the combination of (a), (b) and (c) is effective in the treatment of cancer. In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) PF-07104091 monohydrate; and

(b) PF-07220060 or a pharmaceutically acceptable salt thereof.

In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) monohydrate PF-07104091;

(b) PF-07220060 or a pharmaceutically acceptable salt thereof; and

(c) additional anticancer agent.

In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) PF-07104091 monohydrate; and

(b) selective CDK4 inhibitor.

In another aspect, the invention provides a combination for use in the treatment of cancer, comprising:

(a) monohydrate PF-07104091;

(b) a selective CDK4 inhibitor; and

(c) additional anticancer agent.

In another aspect, the invention provides the use of a combination comprising:

(a) PF-07104091 monohydrate; and

(b) PF-07220060 or a pharmaceutically acceptable salt thereof;

where the use of a combination of (a) and (b) is effective in the treatment of cancer.

In another aspect, the invention provides the use of a combination comprising:

(a) monohydrate PF-07104091;

(b) PF-07220060 or a pharmaceutically acceptable salt thereof; and

(c) additional anticancer agent;

where the use of a combination of (a), (b) and (c) is effective in treating cancer.

In another aspect, the invention provides the use of a combination comprising:

(a) PF-07104091 monohydrate; and

(b) selective CDK4 inhibitor;

where the use of a combination of (a) and (b) is effective in the treatment of cancer.

In another aspect, the invention provides the use of a combination comprising:

(a) monohydrate PF-07104091;

(b) a selective CDK4 inhibitor; and

(c) additional anticancer agent.

where the use of a combination of (a), (b) and (c) is effective in treating cancer.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is selected from the group consisting of breast cancer, prostate cancer, lung cancer (including non-small cell lung cancer, NSCLC, and small cell lung cancer, SCLC), liver cancer (including hepatocellular carcinoma, HCC), kidney cancer (including renal cell carcinoma, RCC), bladder cancer (including urothelial carcinomas such as upper tract urothelial carcinoma, UUTUC), ovarian cancer (including epithelial ovarian cancer, EOC), peritoneal cancer (including primary peritoneal cancer, PPC), fallopian tube cancer, cervical cancer, uterine cancer (including endometrial cancer), pancreatic cancer, gastric cancer, colorectal cancer, esophageal cancer, head and neck cancer (including squamous cell carcinoma of the head and neck (SCCHN), thyroid cancer, and salivary gland cancer. glands), testicular cancer, adrenal cancer, skin cancer (including basal cell carcinoma and melanoma), brain cancer (including astrocytoma, meningioma, and glioblastoma), sarcomas (including osteosarcoma and liposarcoma), and lymphomas (including mantle cell lymphoma, MCL).

In some such embodiments, the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, uterine cancer, colon cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, and gastric cancer.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is characterized by amplification or overexpression of cyclin E1 (CCNE1) and/or cyclin E2 (CCNE2). In some such embodiments, the cancer is characterized by amplification or overexpression of cyclin E1 (CCNE1).

Examples of cyclin E-predominant cancers include, but are not limited to, ovarian cancer, breast cancer, liver cancer, stomach cancer, esophageal cancer, bladder cancer, uterine cancer, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), colon cancer, prostate cancer, or pancreatic cancer.

The retinoblastoma susceptibility gene (RB1) was the first tumor suppressor gene to be defined molecularly. The retinoblastoma gene product, RB, is frequently mutated or deleted in retinoblastoma and osteosarcoma, and is also mutated or deleted with varying frequencies in other tumor types, such as prostate cancer (including prostate neuroendocrine carcinoma), breast cancer (including triple-negative breast cancer, TNBC), lung cancer (including small cell lung cancer, SCLC, and non-small cell lung cancer, NSCLC), liver cancer, bladder cancer, ovarian cancer, uterine cancer, cervical cancer, gastric cancer, esophageal cancer, head and neck cancer, glioblastoma, and lymphoma. In human cancer, RB function can be impaired through neutralization by a binding protein (e.g., human papillomavirus E7 protein in cervical carcinoma; Ishiji, T, 2000[0021], J Dermatol., 27: 73–86) or dysregulation of pathways ultimately responsible for its phosphorylation.

The "RB pathway" refers to the entire molecular signaling pathway that includes the retinoblastoma protein (RB) and other proteins/protein families in this pathway, including, but not limited to, CDK, E2f, atypical protein kinase C, and Skp2. Inactivation of the RB pathway often results from disruption of p16INK4a, cyclin D1, and CDK4.

The terms "RB+," "RB plus," "RB-positive," or "RB-positive" can be used to describe cells expressing detectable amounts of functional RB protein. RB-positive includes both wild-type and non-mutant RB protein. Wild-type RB (RB-WT) is generally understood to refer to the form of RB protein that is normally present in the relevant population and that has the function currently attributed to this protein. RB-positive cells can be cells that contain a functional RB gene. RB-positive cells can also be cells that can encode detectable RB protein function.

The terms "RB-," "RB minus," "RB-deficient," or "RB-negative" describe several cell types in which RB function is impaired, including cells that do not produce detectable amounts of functional RB protein. RB-negative cells may be cells that lack a functional RB gene. RB-negative cells may also be cells that can encode the RB protein but in which the protein does not function properly.

In some embodiments of each of the methods and uses described herein, the cancer is characterized as wild-type retinoblastoma (RB-WT). In some embodiments of each of the methods and uses described herein, the cancer is characterized as RB-positive or RB-excessive. Such RB-positive or RB-excessive cancers contain at least some functional retinoblastoma genes. In some embodiments, such RB-WT, RB-positive, or RB-excessive cancers are characterized as RB1-WT, RB1-positive, or RBI-excessive cancers.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is characterized as RB-negative or RB-deficient. Such RB-negative or RB-deficient cancers may be characterized by loss-of-function mutations, which may encode missense mutations (i.e., encoding an incorrect amino acid) or nonsense mutations (i.e., encoding a stop codon). Alternatively, such RB-negative cancers may be characterized by a deletion of all or part of the retinoblastoma gene. In some embodiments, such RB-negative or RB-deficient cancers are characterized as RBI-negative or RBI-deficient.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is advanced or metastatic cancer.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is difficult to treat, i.e., the cancer does not respond at all to treatment with a therapeutic agent or class (including a standard agent or class) or initially responds but begins to grow again within a very short period of time.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is resistant to a therapeutic agent or class (including a standard agent or class). In some embodiments of each of the methods, combinations, and uses described herein, the cancer is characterized by innate or acquired resistance to a therapeutic agent or class (including a standard agent or class).

In some embodiments of each of the methods, combinations, and uses described herein, the compound of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, and the compound of Formula (II), or a pharmaceutically acceptable salt or solvate thereof, are administered sequentially, successively, or simultaneously. In some embodiments of each of the methods, combinations, and uses described herein, PF-07104091, or a pharmaceutically acceptable salt or solvate thereof, and PF-07220060, or a pharmaceutically acceptable salt or solvate thereof, are administered sequentially, successively, or simultaneously. In some embodiments of each of the methods, combinations, and uses described herein, PF-07104091 and the selective CDK4 inhibitor are administered sequentially, successively, or simultaneously.

In preferred embodiments of the methods, combinations, and uses described herein, the cancer is breast cancer. In some embodiments of each of the breast cancer subtypes described herein, the breast cancer is advanced or metastatic breast cancer.

In some embodiments, the breast cancer is hormone receptor positive (HR+), i.e., the breast cancer is estrogen receptor positive (ER+) and/or progesterone receptor positive (PR+). In some embodiments, the breast cancer is hormone receptor negative (HR-), i.e., the breast cancer is estrogen receptor negative (ER-) and progesterone receptor negative (PR-).

In some embodiments, where the cancer is HR+ breast cancer, the methods, combinations, and uses described herein further include an additional anticancer agent, wherein the additional anticancer agent is an endocrine therapeutic agent. In some such embodiments, the endocrine therapeutic agent is an aromatase inhibitor, SERD, or SERM. In preferred embodiments, the endocrine therapeutic agent is letrozole or fulvestrant.

In some embodiments, the breast cancer is human epidermal growth factor receptor 2 negative (HER2-). In some embodiments, the breast cancer is human epidermal growth factor receptor 2 positive (HER2+).

In some embodiments, breast cancer is associated with the BRCA1 or BRCA2 gene.

In preferred embodiments, the breast cancer is HR+/HER2- breast cancer. In some such embodiments, the HR+/HER2- breast cancer is resistant to treatment with a CDK4/6 inhibitor or progresses with treatment with a CDK4/6 inhibitor. In some such embodiments, the HR+/HER2- breast cancer is resistant to treatment with a CDK4/6 inhibitor, such as palbociclib or a pharmaceutically acceptable salt thereof. In some embodiments, the HR+/HER2- breast cancer is characterized by amplification or overexpression of cyclin E1 (CCNE1) and/or cyclin E2 (CCNE2). In some embodiments, the HR+/HER2- breast cancer is characterized by amplification or overexpression of cyclin E1 (CCNE1). In some embodiments of each of the above, the HR+/HER2- breast cancer is advanced or metastatic HR+/HER2- breast cancer.

In some embodiments, the breast cancer is HR+/HER2+ breast cancer. In other embodiments, the breast cancer is HR-/HER2+ breast cancer. In some embodiments, wherein the breast cancer is HER2+, the methods, combinations, and uses described herein further include an additional anti-cancer agent, wherein the additional anti-cancer agent is an agent targeting HER2 (e.g., trastuzumab emtansine, fam-trastuzumab deruxtecan, pertuzumab, lapatinib, neratinib, or tucatinib) or an agent targeting the PI3K/AKT molecular pathway (e.g., ipatasertib).

In other embodiments, the breast cancer is triple-negative breast cancer (TNBC), i.e., the breast cancer is ER-, PR-, and HER2-. In some embodiments, TNBC is difficult to treat or resistant to treatment with a CDK4/6 inhibitor, such as palbociclib or a pharmaceutically acceptable salt thereof. In some embodiments, TNBC is characterized by amplification or overexpression of cyclin E1 (CCNE1) and/or cyclin E2 (CCNE2). In some such embodiments, TNBC is characterized by amplification or overexpression of cyclin E1 (CCNE1). In some embodiments, TNBC is advanced or metastatic TNBC.

In some embodiments of each of the above, the breast cancer is difficult to treat with one or more standard-of-care agents. In some embodiments of each of the above, the breast cancer is resistant to treatment with one or more standard-of-care agents.

In some such embodiments, the breast cancer is difficult to treat or is resistant to treatment with endocrine therapeutic agents, such as aromatase inhibitors, SERDs, or SERMs. In some embodiments, the breast cancer is difficult to treat or is resistant to, or progresses on, treatment with a CDK4/6 inhibitor. In other embodiments, the breast cancer is difficult to treat or is resistant to, or progresses on, treatment with antitumor chemotherapeutic agents, such as platinum agents, taxanes, anthracyclines, or antimetabolites.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is lung cancer. In some such embodiments, the lung cancer is advanced or metastatic lung cancer.

In some such embodiments, the lung cancer is small cell lung cancer (SCLC). In some such embodiments, the SCLC is characterized by loss of retinoblastoma (RB) function. In some such embodiments, the SCLC is advanced or metastatic SCLC. In some such embodiments, the SCLC is advanced or metastatic SCLC characterized by loss of retinoblastoma (RB) function.

In some embodiments of the methods and uses described herein, the cancer is SCLC. In some such embodiments, the SCLC is Rb-negative or Rb-deficient.

In some embodiments of the methods and uses described herein, the cancer is NSCLC. In some such embodiments, NSCLC is characterized by amplification or overexpression of cyclin E1 (CCNE1) and/or cyclin E2 (CCNE2). In some embodiments, NSCLC is characterized by amplification or overexpression of cyclin E1 (CCNE1). In some such embodiments, NSCLC is advanced or metastatic NSCLC. In some such embodiments, NSCLC is advanced or metastatic NSCLC characterized by amplification or overexpression of cyclin E1 (CCNE1). In other embodiments, NSCLC is lung squamous cell carcinoma (LUSC) or lung adenocarcinoma (LUAD). In a preferred embodiment, the lung cancer is LUAD. Single-gene oncogenic drivers of lung adenocarcinoma include, but are not limited to, EGFR, BRAF, and KRAS. Approximately 25% of lung adenocarcinomas are driven by KRAS, which may include segments driven by KRAS G12C and non-G12C, such as tumors driven by G12A, G12D, G12V, G13D, and L19F.

In some embodiments, the cancer is lung cancer, including SCLC or NSCLC, and the methods, combinations, and uses described herein further include an additional anti-cancer agent.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is ovarian cancer, peritoneal cancer, or fallopian tube cancer (FTC). In some such embodiments, the ovarian cancer is epithelial ovarian cancer (EOC). In some such embodiments, the peritoneal cancer is primary peritoneal carcinomatosis (PPC). In some embodiments, the cancer is epithelial ovarian cancer (EOC), primary peritoneal carcinomatosis (PPC), or fallopian tube cancer (FTC). In some embodiments, the ovarian cancer is persistent, difficult to treat, or recurrent ovarian cancer. In some such embodiments, the ovarian cancer is platinum-resistant ovarian cancer. In some such embodiments, the cancer is advanced or metastatic ovarian cancer. In some such embodiments, the cancer is advanced or metastatic platinum-resistant ovarian cancer. In some such embodiments, the cancer is advanced or metastatic EOC, RMS, or FTC. In some such embodiments, the cancer is platinum-resistant advanced or metastatic EOC, RMS, or FTC.

In some embodiments, each of the methods, combinations, and uses described herein further comprises an additional anticancer agent. In some embodiments, the additional anticancer agent is a standard of care agent for the given cancer type. In some embodiments, each of the methods, combinations, and uses described herein further comprises an additional anticancer agent, wherein the compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof, the compound of Formula (II) or a pharmaceutically acceptable salt or solvate, and the additional anticancer agent are administered sequentially, consecutively, or simultaneously.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is breast cancer (including HR+ or HR+/HER2- breast cancer), and the methods, combinations, and uses further include an additional anti-cancer agent. In some embodiments, the additional anti-cancer agent is an endocrine therapeutic agent. In some such embodiments, the endocrine therapeutic agent is an aromatase inhibitor, SERD, or SERM. In some embodiments, the endocrine therapeutic agent is an aromatase inhibitor. In some such embodiments, the aromatase inhibitor is selected from the group consisting of letrozole, anastrozole, and exemestane. In some preferred embodiments, the aromatase inhibitor is letrozole. In some embodiments, the endocrine therapeutic agent is a SERD. In some such embodiments, the SERD is selected from the group consisting of fulvestrant, elacestrant (RAD-1901, Radius Health), SAR439859 (Sanofi), RG6171 (Roche), AZD9833 (AstraZeneca), AZD94 96 (AstraZeneca), rintodestrant (G1 Therapeutics), ZN-c5 (Zentalis), LSZ102 (Novartis), D-0502 (Inventisbio), LY3484356 (Lilly), and SHR9549 (Jiansu Hengrui Medicine). In some preferred embodiments, the SERD is fulvestrant. In some embodiments, the endocrine therapeutic agent is a SERM. In some such embodiments, the SERM is selected from the group consisting of tamoxifene, raloxifene, toremifene, lasofoxifene, bazedoxifene, and afinoxifene. In some such embodiments, the SERM is tamoxifen or raloxifene.

In some embodiments of each of the methods, combinations, and uses described herein, the cancer is breast cancer (including HR+ or HR+/HER2- breast cancer), and the methods, combinations, and uses further include an additional anti-cancer agent, wherein the compound of Formula (I) is PF-07104091, the compound of Formula (II) is PF-07220060 or a pharmaceutically acceptable salt thereof, and the additional anti-cancer agent is an endocrine therapeutic agent, wherein the endocrine therapeutic agent is an aromatase inhibitor, SERD, or SERM. In some such embodiments, the endocrine therapeutic agent is letrozole or fulvestrant.

Pharmaceutical Compositions, Medicaments and Kits The present invention further relates to pharmaceutical compositions, medicaments and kits containing a compound of Formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein:

R ¹ is -L-(5-6-membered heteroaryl) or -L-(phenyl), wherein said 5-6-membered heteroaryl or phenyl is optionally substituted with one to three R ³ ;

R ² is C ₁ -C ₆ alkyl or C ₃ -C ₇ cycloalkyl, wherein said C ₃ -C ₇ cycloalkyl is optionally substituted with C ₁ -C ₄ alkyl;

L represents a bond or methylene; and

each R ³ is independently C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or SO ₂ -C ₁ -C 4 alkyl, wherein each C ₁ -C ₄ alkyl is optionally substituted with F, OH, or C ₁ -C ₄ alkoxy.

In another aspect, the invention provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof, a selective CDK4 inhibitor or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In some embodiments of this aspect, the pharmaceutical composition further comprises an additional anticancer agent (e.g., an endocrine therapeutic agent).

In another aspect, the invention provides a first pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof and a pharmaceutically acceptable carrier or excipient, and a second pharmaceutical composition comprising a compound of Formula (II) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, wherein the first and second pharmaceutical compositions are administered sequentially, successively, or simultaneously. Some embodiments of this aspect further include a third pharmaceutical composition comprising an additional anticancer agent (e.g., an endocrine therapeutic agent) and a pharmaceutically acceptable carrier or excipient, wherein the first, second, and third pharmaceutical compositions are administered sequentially, successively, or simultaneously.

In another aspect, the invention provides a combination comprising a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof, and a selective CDK4 inhibitor or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for the treatment of cancer in a subject.

In another aspect, the invention provides the use of a combination comprising PF-07104091 or a pharmaceutically acceptable solvate thereof and PF-07220060 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of cancer in a subject. In some embodiments of these aspects, the combination further comprises an additional anticancer agent (e.g., an endocrine therapeutic agent) for use in the manufacture of the medicament.

In another aspect, the invention provides a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof for use in the manufacture of a medicament for the treatment of cancer, wherein the medicament is adapted for use in combination with a compound of Formula (II) or a pharmaceutically acceptable salt thereof. In another aspect, the invention provides a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof for use in the manufacture of a medicament for the treatment of cancer, wherein the medicament is adapted for use in combination with a compound of Formula (II) or a pharmaceutically acceptable salt thereof, and an additional anticancer agent, for example, an endocrine therapeutic agent.

In preferred embodiments of each of the pharmaceutical compositions, kits, and medicaments described herein, the compound of Formula (I) is PF-07104091 or a monohydrate thereof. In preferred embodiments of each of the pharmaceutical compositions, kits, and medicaments described herein, the compound of Formula (II) is PF-07220060 or a pharmaceutically acceptable salt thereof.

In particularly preferred embodiments of each of the pharmaceutical compositions, kits, and medicaments described herein, the compound of Formula (I) is PF-07104091 monohydrate and the compound of Formula (II) is PF-07220060.

In another aspect, the invention provides a kit comprising a first container, a second container, and a package insert, wherein the first container contains at least one dose of a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof; the second container contains at least one dose of a compound of Formula (II) or a pharmaceutically acceptable salt thereof; and the package insert contains instructions for treating cancer in a subject using a medicament. In some embodiments, the kit further comprises a third container, wherein the third container contains at least one dose of an additional anticancer agent (e.g., an endocrine therapeutic agent); and the package insert contains instructions for treating cancer in a subject using a medicament.

In embodiments of pharmaceutical compositions, medicaments, and kits containing an additional anticancer agent, the additional anticancer agent is an endocrine therapeutic agent, such as an aromatase inhibitor, SERD, or SERM. In some such embodiments, the endocrine therapeutic agent is letrozole or fulvestrant.

The pharmaceutical compositions, medicaments, and kits described herein may be useful for treating the cancer described above in relation to the methods, combinations, and uses disclosed herein. In some embodiments, the pharmaceutical compositions, medicaments, and kits may be useful for treating a cancer selected from the group consisting of breast cancer (including HR+/HER2-, HR+/HER2+, HR-/HER2+, or TNBC), lung cancer (including SCLC or NSCLC), ovarian cancer (including EOC), peritoneal cancer (including RMS), fallopian tube cancer (including FTC), bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, and gastric cancer.

Dosage forms and treatment regimens

Each therapeutic agent included in the combinations, dosage regimens, methods, and uses described herein can be administered either alone or as a medicament (also referred to herein as a pharmaceutical composition) that includes the therapeutic agent and one or more pharmaceutically acceptable carriers, excipients, or diluents, in accordance with pharmaceutical practice.

As used herein, the terms "combination" or "combination therapy" refer to the administration of two or more therapeutic agents in a combination therapy, either as a compound or as a pharmaceutical composition. Combination therapy may be administered sequentially, consecutively, or simultaneously.

In some embodiments, the effective amount of PF-07104091 when used daily in combination with PF-07220060 (i.e., the total daily dose) is from about 100 mg to about 500 mg per day, from about 100 mg to about 450 mg per day, from about 100 mg to about 400 mg per day, from about 100 mg to about 375 mg per day, from about 100 mg to about 350 mg per day, from about 100 mg to about 325 mg per day, from about 100 mg to about 300 mg per day, from about 100 mg to about 275 mg per day, from about 100 mg to about 250 mg per day, from about 100 mg to about 225 mg per day, from about 100 mg to about 200 mg per day, from about 100 mg to about 175 mg per day, from about 100 mg to about 150 mg per day, from about 150 mg to about 500 mg per day, from about 150 mg to about 450 mg per day, from about 150 mg to about 400 mg per day, from about 150 mg to about 375 mg per day, from about 150 mg to about 350 mg per day, from about 150 mg to about 325 mg per day, from about 150 mg to about 300 mg per day, from about 150 mg to about 275 mg per day, from about 150 mg to about 250 mg per day, from about 150 mg to about 225 mg per day, from about 150 mg to about 200 mg per day, from about 200 mg to about 500 mg per day, from about 200 mg to about 450 mg per day, from about 200 mg to about 400 mg per day, from about 200 mg to about 375 mg per day, from about 200 mg to about 350 mg per day, from about 200 mg to about 325 mg per day, from about 200 mg to about 300 mg per day, from about 200 mg to about 275 mg per day, or from about 200 mg to about 250 mg per day.

In some embodiments, the effective amount of PF-07104091 when administered QD in combination with PF-07220060 is from about 100 mg to about 500 mg QD, from about 100 mg to about 450 mg QD, from about 100 mg to about 400 mg QD, from about 100 mg to about 375 mg QD, from about 100 mg to about 350 mg QD, from about 100 mg to about 325 mg QD, from about 100 mg to about 300 mg QD, from about 100 mg to about 275 mg QD, from about 100 mg to about 250 mg QD, from about 100 mg to about 225 mg QD, from about 100 mg to about 200 mg QD, from about 100 mg to about 175 mg QD, from about 100 mg to about 150 mg QD, about 150 mg to about 500 mg QD, about 150 mg to about 450 mg QD, about 150 mg to about 400 mg QD, about 150 mg to about 375 mg QD, about 150 mg to about 350 mg QD, about 150 mg to about 325 mg QD, about 150 mg to about 300 mg QD, about 150 mg to about 275 mg QD, about 150 mg to about 250 mg QD, about 150 mg to about 225 mg QD, about 150 mg to about 200 mg QD, about 200 mg to about 500 mg QD, about 200 mg to about 450 mg QD, about 200 mg to about 400 mg QD, from about 200 mg to about 375 mg QD, from about 200 mg to about 350 mg QD, from about 200 mg to about 325 mg QD, from about 200 mg to about 300 mg QD, from about 200 mg to about 275 mg QD, from about 200 mg to about 250 mg QD.

In preferred embodiments, the effective amount of PF-07104091 when administered BID in combination with PF-07220060 is from about 50 mg to about 250 mg BID, from about 50 mg to about 225 mg BID, from about 50 mg to about 200 mg BID, from about 50 mg to about 175 mg BID, from about 50 mg to about 150 mg BID, from about 50 mg to about 125 mg BID, from about 50 mg to about 100 mg BID, from about 75 mg to about 250 mg BID, from about 75 mg to about 225 mg BID, from about 75 mg to about 200 mg BID, from about 75 mg to about 175 mg BID, from about 75 mg to about 150 mg BID, from about 75 mg to about 125 mg BID, from about 75 mg to about 100 mg BID, from about 100 mg to about 250 mg BID, from about 100 mg to about 225 mg BID, from about 100 mg to about 200 mg BID, from about 100 mg to about 175 mg BID, from about 100 mg to about 150 mg BID, or from about 100 mg to about 125 mg BID,

In some embodiments, PF-07104091, when used in combination with PF-07220060, is administered at a dose of about 100 mg per day, about 150 mg per day, about 200 mg per day, about 250 mg per day, about 300 mg per day, about 350 mg per day, or about 400 mg per day.

In some embodiments, PF-07104091, when used in combination with PF-07220060, is administered at a dose of about 100 mg QD, about 150 mg QD, about 200 mg QD, about 250 mg QD, about 300 mg QD, about 350 mg QD, or about 400 mg QD.

In some embodiments, PF-07104091, when used in combination with PF-07220060, is administered at a dose of about 50 mg BID, about 75 mg BID, about 100 mg BID, about 125 mg BID, about 150 mg BID, about 175 mg Twice a day, or about 200 mg BID.

In preferred embodiments, PF-07104091 is administered on a twice daily (BID) schedule. In preferred embodiments, the effective amount of PF-07104091 when used in combination with PF-07220060 is about 75 mg BID, about 100 mg BID, about 125 mg BID, or about 150 mg BID.

In some embodiments, a therapeutically effective amount of PF-07220060 when used in combination with PF-07104091 is from about 200 mg to about 1000 mg per day (i.e., a total daily dose), such as from about 200 mg to about 500 mg, from about 200 mg to about 450 mg, from about 200 mg to about 400 mg, from about 200 mg to about 350 mg, from about 200 mg to about 300 mg, from about 400 mg to about 950 mg, from about 400 mg to about 900 mg, from about 400 mg to about 850 mg, from about 400 mg to about 800 mg, from about 500 mg to about 950 mg, from about 500 mg to about 900 mg, from about 500 mg to about 850 mg, from about 500 mg to about 800 mg, from about 600 mg to about 950 mg, from about 600 mg to about 900 mg, from about 600 mg to about 850 mg, or from about 600 mg to about 800 mg per day.

In some embodiments, PF-07220060, when used in combination with PF-07104091, is administered at doses of from about 200 mg to about 1000 mg once per day (QD), such as from about 200 mg to about 500 mg QD, from about about 200 mg to about 450 mg QD, from about 200 mg to about 400 mg QD, from about 200 mg to about 350 mg QD, from about 200 mg to about 300 mg QD, from about 400 mg to about 950 mg QD, from about 400 mg to about 900 mg QD, from about 400 mg to about 850 mg QD, from about 400 mg to about 800 mg QD, from about 500 mg to about 950 mg QD, from about 500 mg to about 900 mg QD, from about 500 mg to about 850 mg QD, from about 500 mg to about 800 mg QD, from about 600 mg to about 950 mg QD, from about 600 mg to about 900 mg QD, from about 600 mg to about 850 mg QD, or from about 600 mg to about 800 mg QD.

In some embodiments, a therapeutically effective amount of PF-07220060 when used in combination with PF-07104091 is from about 100 mg to about 500 mg twice daily (BID), such as from about 200 mg to about 500 mg BID, from about 250 mg to about 500 mg BID, from about 300 mg to about 500 mg BID, from about 350 mg to about 500 mg BID, from about 400 mg to about 500 mg BID, from about 200 mg to about 450 mg BID, from about 250 mg to about 450 mg BID, from about 300 mg to about 450 mg BID, from about 350 mg to about 450 mg BID, from about 400 mg to about 450 mg BID, from about 200 mg to about 400 mg BID, from about 250 mg to about 400 mg BID, from about 300 mg to about 400 mg BID, or from about 350 mg to about 400 mg BID.

In some embodiments, PF-07220060 is administered to a subject when used in combination with PF-07104091 at a dose of about 200 mg to about 1000 mg per day, such as at daily doses of about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or about 1000 mg.

In some embodiments, PF-07220060 is administered at doses of about 200 mg QD, about 300 mg QD, about 400 mg QD, about 500 mg QD, about 600 mg QD, about 700 mg QD, about 800 mg QD, about 900 mg QD, or about 1000 mg QD.

In preferred embodiments, PF-07220060, when used in combination with PF-07104091, is administered at a dose of about 100 mg BID, about 200 mg BID, about 300 mg BID, about 400 mg BID, or about 500 mg BID.

In some embodiments, the subject is administered PF-07104091 and PF-07220060 at a dose of any of the effective amounts disclosed herein.

In some embodiments, the subject is administered about 75 mg of PF-07104091 BID and about 100 mg of PF-07220060 BID. In some embodiments, the subject is administered about 150 mg of PF-07104091 BID and about 100 mg of PF-07220060 BID. In some embodiments, the subject is administered about 225 mg of PF-07104091 BID and about 100 mg of PF-07220060 BID. In some embodiments, the subject is administered about 75 mg of PF-07104091 BID and 200 mg of PF-07220060 BID. In some embodiments, the subject is administered about 150 mg of PF-07104091 BID and about 200 mg of PF-07220060 BID. In some embodiments, the subject is administered about 225 mg of PF-07104091 BID and about PF-07220060. In some embodiments, the subject is administered about 75 mg of PF-07104091 BID and about 300 mg of PF-07220060 BID. In some embodiments, the subject is administered about 150 mg of PF-07104091 BID and about 300 mg of PF-07220060 BID. In some embodiments, the subject is administered about 225 mg of PF-07104091 BID and about 300 mg of PF-07220060 BID.

The amounts of PF-07104091 and PF-07220060 administered may be increased or decreased depending on the subject's weight, age, health, gender, or medical condition. A person skilled in the art will be able to determine the appropriate dose for a subject based on this description.

When administering combination therapy including PF-07104091 and PF-07220060, the agents may be administered in the same treatment cycle or in different treatment cycles.

PF-07104091 and PF-07220060 can be administered in treatment cycles with or without rest periods between treatment cycles. A treatment cycle can be approximately 7 days, approximately 14 days, approximately 21 days, approximately 28 days, approximately 35 days, and so on, or any days in between. The rest period can be one or more days (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and so on), one week, several weeks (e.g., 2 weeks, 3 weeks, and so on), or any days in between (e.g., 1 week and 3 days).

In some embodiments, PF-07104091 is administered continuously without a rest period between treatment cycles (i.e., continuous treatment until discontinued). In some embodiments, PF-07104091 is administered throughout a treatment cycle (e.g., about 28 days) with or without a rest period. In some embodiments, PF-07104091 is administered for about 28 days with a rest period of about one week. PF-07104091 can be administered for at least about 7 days, about 14 days, about 21 days, about 28 days, about 2 months, about 3 months, about 12 months, about 24 months, or more. In a preferred embodiment, PF-07104091 is administered continuously throughout a 28-day treatment cycle without a rest period.

In some embodiments, PF-07220060 is administered continuously without a rest period between treatment cycles (i.e., continuous treatment until discontinued). In some embodiments, PF-07220060 is administered throughout a treatment cycle (e.g., about 28 days) with or without a rest period. In some embodiments, PF-07220060 is administered for about 28 days with a rest period of about one week. 07220060 can be administered for at least about 7 days, about 14 days, about 21 days, about 28 days, about 2 months, about 3 months, about 12 months, about 24 months, or more. In a preferred embodiment, 07220060 is administered continuously throughout a 28-day treatment cycle without a rest period.

Pharmaceutical compositions can be administered with or without food.

Pharmaceutical compositions can be administered by one or more routes deemed appropriate by those skilled in the art, depending on the dosage form. Pharmaceutical compositions can be administered with or without food. Drug formulations are discussed in Remington's Pharmaceutical Sciences, 18th Ed., (1995) Mack Publishing Co., Easton, Pa. Other examples of drug formulations can be found in Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, Vol 3, 2nd Ed., New York, NY. If the compound is administered orally, it can be formulated as a pill, capsule, tablet, etc. with a pharmaceutically acceptable carrier, glidant, or excipient.

The pharmaceutical compositions may be in one or more dosage forms (e.g., capsule, tablet, powder, or liquid).

In some embodiments of each of the methods, combinations, and uses described herein, the combination therapy is administered to a subject who is previously naive, i.e., has not previously received treatment.

In some embodiments of each of the methods, combinations, and uses described herein, the combination therapy is administered to a subject who has failed to achieve a durable response following prior therapy with a biotherapeutic or chemotherapeutic agent, i.e., who has undergone treatment.

In some embodiments of each of the methods, combinations, and uses described herein, the combination therapy may be administered to a subject who has previously been treated with chemotherapy, radiation therapy, and/or surgical resection.

In some embodiments of each of the methods, combinations, and uses of combination therapy described herein, the subject has previously been treated with a CDK4/6 inhibitor. In some embodiments, the CDK4/6 inhibitor is palbociclib.

In some embodiments of each of the methods, combinations, and uses described herein, the invention relates to neoadjuvant therapy, adjuvant therapy, first-line therapy, second-line therapy, or third-line or later therapy, in each case for the treatment of cancer as further described herein. In each of the above embodiments, the cancer may be localized, advanced, or metastatic, and the intervention may occur at any point along the disease continuum (i.e., at any stage of the cancer).

Dosage regimens can be adjusted to provide the optimal desired response. For example, the therapeutic agent of the combination therapy disclosed herein can be administered as a single bolus, as several divided doses administered over time, or the dose can be proportionally reduced or increased, as indicated, depending on the requirements of the therapeutic situation. It may be particularly advantageous to formulate the therapeutic agent in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form, as used herein, refers to physically discrete units suitable as unitary dosages for the mammals to be treated; each unit contains a predetermined quantity of the active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification of unit dosage forms may be dictated by and directly dependent on (a) the unique characteristics of the chemotherapeutic agent and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the field of formulation of such active compound for the treatment of susceptibility in individuals.

In some embodiments, the endocrine therapeutic agent is letrozole, which can be administered orally at a dose of 2.5 mg per day. In some embodiments, the endocrine therapeutic agent is fulvestrant, which can be administered intramuscularly as one or two injections at a dose of 250 mg or 500 mg, respectively, on days 1, 15, and 29 of the first month, and then once per month thereafter.

In some embodiments, the compound of Formula (I) and the compound of Formula (II) are administered in an intermittent dosing regimen. In other embodiments, the compound of Formula (I) and the compound of Formula (II) are administered in a continuous dosing regimen.

In other embodiments, one of the compounds of Formula (I) and the compound of Formula (II) is administered on an intermittent dosing schedule (e.g., on a 2/1 week or 3/1 week schedule), and the other is administered on a continuous dosing schedule. In some such embodiments, the compound of Formula (I) is administered on an intermittent dosing schedule, and the compound of Formula (II) is administered on a continuous dosing schedule. In other such embodiments, the compound of Formula (I) is administered on a continuous dosing schedule, and the compound of Formula (II) is administered on an intermittent dosing schedule.

In some embodiments disclosed herein, the compound of Formula (I) and the compound of Formula (II) are dosed in amounts that are together effective in treating cancer.

In some embodiments disclosed herein, the compound of Formula (I) and the compound of Formula (II) are dosed in amounts that together are synergistic.

In some embodiments disclosed herein, the compound of Formula (I) and the compound of Formula (II) are dosed in amounts that together are additive.

In preferred embodiments of each of the above, the compound of Formula (I) is PF-07104091 or a monohydrate thereof, and the compound of Formula (II) is PF-07220060 or a pharmaceutically acceptable salt thereof. In particularly preferred embodiments of each of the above, the compound of Formula (I) is PF-07104091 monohydrate, and the compound of Formula (II) is PF-07220060.

Pharmaceutical Compositions and Routes of Administration "Pharmaceutical composition" refers to a mixture of one or more therapeutic agents described herein, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, as an active ingredient and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and/or excipients.

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that is not significantly irritating to the body and does not interfere with the biological activity and properties of the active compound or therapeutic agent.

A pharmaceutically acceptable carrier may include any conventional pharmaceutical carrier or excipient. The choice of carrier and/or excipient will depend largely on factors such as the particular route of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

Suitable pharmaceutical carriers include inert diluents or fillers, water, and various organic solvents (such as hydrates and solvates). Pharmaceutical compositions may optionally contain additional ingredients such as flavorings, binding agents, excipients, and the like. Thus, for oral administration, tablets can be used containing various excipients, such as citric acid, along with various disintegrants, such as starch, alginic acid, and certain complex silicates, and with binding agents, such as sucrose, gelatin, and acacia. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols. Additionally, lubricating agents, such as magnesium stearate, sodium lauryl sulfate, and talc, are often used for tableting. Solid compositions of a similar type can also be used in soft and hard-filled gelatin capsules. Thus, non-limiting examples of materials include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the active compound contained therein can be combined with various sweeteners or flavorings, colorings, and, optionally, emulsifiers or suspending agents, along with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.

The pharmaceutical composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, solution or suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream, or for rectal administration as a suppository.

Typical parenteral dosage forms include solutions or suspensions of the active compound in a sterile aqueous solution, such as aqueous propylene glycol or dextrose. If desired, such dosage forms can be appropriately buffered.

The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise quantities.

Pharmaceutical compositions suitable for delivering the therapeutic agents of the combination therapy disclosed herein, and methods for their preparation, will be readily apparent to those skilled in the art. Such compositions and methods for their preparation can be found, for example, in "Remington's Pharmaceutical Sciences," 19th Edition (Mack Publishing Company, 1995), the disclosure of which is incorporated herein by reference in its entirety.

The therapeutic agents of the combination therapy described herein can be administered orally. Oral administration can involve swallowing, so that the therapeutic agent enters the gastrointestinal tract, or buccal or sublingual administration can be used, whereby the therapeutic agent enters the bloodstream directly from the mouth.

Formulations suitable for oral administration include solid formulations such as tablets, capsules containing particles, liquids or powders, lozenges (including liquid-filled), chewable tablets, multi- and nanoparticulates, gels, solid solutions, liposomes, films (including mucoadhesive), capsules, sprays and liquid formulations.

Liquid formulations include suspensions, solutions, syrups, and elixirs. Such formulations can be used as fillers in soft or hard capsules and typically comprise a carrier, such as water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, and one or more emulsifying and/or suspending agents. Liquid formulations can also be prepared by reconstitution of a solid, such as from a sachet.

The combination therapy therapeutic agents disclosed herein may also be used in rapidly dissolving and rapidly disintegrating dosage forms, such as those described in Expert Opinion in Therapeutic Patents, 11 (6), 981-986 Liang and Chen (2001), the disclosure of which is incorporated herein by reference in its entirety.

For tablet dosage forms, the therapeutic agent may comprise from 1% to 80% by weight of the dosage form, more commonly from 5% to 60% by weight of the dosage form. In addition to the active agent, tablets typically contain a disintegrating agent. Examples of disintegrating agents include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, sodium croscarmellose, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch, and sodium alginate. Typically, the disintegrating agent may comprise from 1% to 25% by weight, preferably from 5% to 20% by weight, of the dosage form.

Binding agents are commonly used to impart cohesive properties to tablet formulations. Suitable binding agents include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose, and hydroxypropyl methylcellulose. Tablets may also contain diluents such as lactose (monohydrate, lyophilized monohydrate, anhydrous, and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch, and dibasic calcium phosphate dihydrate.

Tablets may also optionally include surfactants, such as sodium lauryl sulfate and polysorbate 80, and glidants, such as silicon dioxide and talc. Surfactants, if present, typically comprise 0.2% to 5% by weight of the tablet, and glidants typically comprise 0.2% to 1% by weight of the tablet.

Tablets also typically contain lubricating agents such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. Lubricating agents are typically present in amounts ranging from 0.25% to 10% by weight, preferably from 0.5% to 3% by weight, of the tablet.

Other traditional ingredients include antioxidants, colors, flavors, preservatives, and taste-masking agents.

Typical tablets may contain up to about 80% by weight of active agent, from about 10% by weight to about 90% by weight of binder, from about 0% by weight to about 85% by weight of diluent, from about 2% by weight to about 10% by weight of disintegrant, and from about 0.25% by weight to about 10% by weight of lubricant.

Tablet blends can be compressed directly or using a roller to form tablets. Alternatively, tablet blends or portions of blends can be wet, dry, or melt granulated, melt solidified, or extruded prior to tableting. The final composition may include one or more layers and may be coated or uncoated; or encapsulated.

The composition of the tablets is discussed in detail in "Pharmaceutical Dosage Forms: Tablets, Vol. 1" by H. Lieberman and L. Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0-8247-6918-X), the disclosure of which is incorporated herein by reference in its entirety.

Solid oral formulations can be formulated with immediate and/or modified release. Modified release formulations include delayed, sustained, pulsed, controlled, targeted, and programmed release.

Suitable modified release formulations are described in U.S. Patent No. 6,106,864. Details of other suitable release technologies, such as high-energy dispersions and osmotic and coated particles, can be found in Verma et al., Pharmaceutical Technology On-line, 25(2), 1-14 (2001). The use of chewing gum to achieve controlled release is described in WO 2000/035298. The disclosures of these references are incorporated herein by reference in their entirety.

The kits described herein may be particularly suitable for administering various dosage forms, such as oral and parenteral, for administering individual compositions at different dosing intervals, or for titrating individual compositions against each other. To facilitate compliance, the kit typically includes instructions for use and may be equipped with a memory aid. The kit may additionally include other materials that may be useful in administering the drug, such as diluents, filters, intravenous infusion bags and catheters, needles and syringes, and the like.

Additional anticancer agents

The methods, combinations, and uses disclosed herein may further include one or more additional anti-cancer agents, such as anti-angiogenesis agents, signal transduction inhibitors, or anti-tumor agents described below, wherein the amounts collectively are effective in treating cancer. In some embodiments of the methods, combinations, and uses disclosed herein, the additional anti-cancer agents may include a palliative care agent. Additional anti-cancer agents may include small molecule therapeutics and pharmaceutically acceptable salts or solvates thereof, therapeutic antibodies, antibody-drug conjugates (ADCs), heterobifunctional protein biodegraders (e.g., proteolysis-targeting chimeras, or PROTACs), or antisense molecules.

In some embodiments, the methods, combinations, and uses disclosed herein further include one or more additional anticancer agents selected from the following:

Anti-angiogenesis agents include, for example, VEGF inhibitors, VEGFR inhibitors, TIE-2 inhibitors, PDGFR inhibitors, angiopoietin inhibitors, PKCβ inhibitors, COX-2 (cyclooxygenase II) inhibitors, integrins (alpha-v/beta-3), MMP-2 (matrix metalloproteinase 2) inhibitors, and MMP-9 (matrix metalloproteinase 9) inhibitors.

Signal transduction inhibitors include, for example, kinase inhibitors (e.g., tyrosine kinase, serine/threonine kinase, or cyclin-dependent kinase inhibitors), proteasome inhibitors, PI3K/AKT/mTOR pathway inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) inhibitors, B-cell lymphoma 2 (BCL2) inhibitors, neurotrophin receptor kinase (NTRK) inhibitors, rearranged during transfection (RET) inhibitors, Notch inhibitors, PARP inhibitors, Hedgehog pathway inhibitors, and selective inhibitors of nuclear export (SINE).

Examples of signal transduction inhibitors include, but are not limited to: acalabrutinib, afatinib, alectinib, alpelisib, axitinib, binimetinib, bortezomib, bosutinib, brigatinib, cabozantinib, carfilzomib, ceritinib, cobimetinib, copanlisib, crizotinib, dabrafenib, dacomitinib, dasatinib, duvelisib, enasidenib, encorafenib, entrectinib, erlotinib, gefitinib, gilteritinib, glasdegib, ibrutinib, idelalisib, imatinib, ipatasertib, ivosidenib, ixazomib, lapatinib, larotrectinib, lenvatinib, lorlatinib, midostaurin, neratinib, nilotinib, niraparib, olaparib, osimertinib, pazopanib, ponatinib, regorafenib, rucaparib, ruxolitinib, sonidegib, sorafenib, sunitinib, talazoparib, trametinib, vandetanib, vemurafenib, venetoclax and vismodegib or pharmaceutically acceptable salts and solvates thereof.

Antineoplastic agents include, for example, alkylating agents, platinum coordination complexes, cytotoxic antibiotics, antimetabolite agents, biological response modifiers, histone deacetylase (HDAC) inhibitors, hormonal agents, monoclonal antibodies, growth factor inhibitors, taxanes, topoisomerase inhibitors, vinca alkaloids, and miscellaneous agents.

Alkylating agents include: altretamine, bendamustine, busulfan, carmustine, chlorambucil, cyclophosphamide, dacarbazine, ifosfamide, lomustine, mechlorethamine, melphalan, procarbazine, streptozocin, temozolomide, thiotepa, and trabectedin.

Platinum coordination complexes include: carboplatin, cisplatin, and oxaliplatin.

Cytotoxic antibiotics include: bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitomycin, mitoxantrone, plicamycin, and valrubicin.

Antimetabolites include: antifolates such as methotrexate, pemetrexed, pralatrexate, and trimetrexate; purine analogues such as azathioprine, cladribine, fludarabine, mercaptopurine, and thioguanine; and pyrimidine analogues such as azacitidine, capecitabine, cytarabine, decitabine, floxuridine, fluorouracil, gemcitabine, and trifluridine/tipracil.

Biological response modifiers include aldesleukin (IL-2), denileukin diftitox, and interferon gamma.

Histone deacetylase inhibitors include belinostat, panobinostat, romidepsin, and vorinostat.

Endocrine therapeutics (i.e., hormone therapy agents) include antiandrogens, antiestrogens, gonadotropin-releasing hormone (GnRH) analogs, and peptide hormones. Examples of antiestrogens include: aromatase inhibitors such as letrozole, anastrozole, and exemestane; SERDs such as fulvestrant, elacestrant (RAD-1901, Radius Health/Menarini), amcenestrant (SAR439859, Sanofi), giredestrant (GDC9545, Roche), RG6171 (Roche), kamisestrant (AZD9833, AstraZeneca), AZD9496 (AstraZeneca), rintodestrant (G1 Therapeutics), ZN-c5

(Zentalis), LSZ102 (Novartis), D-0502 (Inventisbio), LY3484356 (Eli Lilly), SHR9549 (Jiansu Hengrui Medicine); and SERMs such as tamoxifen, raloxifene, toremifene, lasofoxifene, bazedoxifene, afimoxifene. Examples of GnRH analogs include: degarelix, goserelin, histrelin, leuprolide, and triptorelin. Examples of peptide hormones include: lanreotide, octreotide, and pasireotide. Examples of antiandrogens include: abiraterone, apalutamide, bicalutamide, cyproterone, enzalutamide, flutamide, and nilutamide, and their pharmaceutically acceptable salts and solvates.

Monoclonal antibodies include: alemtuzumab, atezolizumab, avelumab, bevacizumab, blinatumomab, brentuximab, cemiplimab, cetuximab, daratumumab, dinutuximab, durvalumab, elotuzumab, gemtuzumab, inotuzumab ozogamicin, ipilimumab, izumab, moxetumomab pasudotox, necitumumab, nivolumab, ofatumumab, olaratumab, panitumumab, pembrolizumab, pertuzumab, ramucirumab, rituximab, tositumomab, and trastuzumab.

Taxanes include: cabazitaxel, docetaxel, paclitaxel, and paclitaxel-albumin-stabilized nanoparticle formulation.

Topoisomerase inhibitors include: etoposide, irinotecan, teniposide, and topotecan.

Vinca alkaloids include vinblastine, vincristine, and vinorelbine, and their pharmaceutically acceptable salts.

Miscellaneous antineoplastic agents include: asparaginase (pegaspargase), bexarotene, eribulin, everolimus, hydroxyurea, ixabepilone, lenalidomide, mitotane, omacetaxine, pomalidomide, tagraxofusp, telerotistat, temsirolimus, thalidomide, and venetoclax.

In some embodiments, the additional anti-cancer agent is selected from the group consisting of: abiraterone acetate; acalabrutinib; ado-trastuzumab emtansine; afatinib dimaleate; afimoxifene; aldesleukin; alectinib; alemtuzumab; alpelisib; amifostine; anastrozole; apalutamide; aprepitant; arsenic trioxide; erwinia chrysanthemi asparaginase; atezolizumab; avapritinib; avelumab; axicabtagene ciloleucel; axitinib; azacitidine; AZD9833 (AstraZeneca); AZD9496 (AstraZeneca); bazedoxifene; belinostat; bendamustine hydrochloride; bevacizumab; bexarotene; bicalutamide; binimetinib; bleomycin sulfate; blinatumomab; bortezomib; bosutinib; brentuximab vedotin; brigatinib; cabazitaxel; cabozantinib-s-malate; calaspargase pegola-mknl; capecitabine; caplacizumab-yhdp; capmatinib hydrochloride; carboplatin; carfilzomib; carmustine; cemiplimab-rwlc; ceritinib; cetuximab; chlorambucil; cisplatin; cladribine; clofarabine; cobimetinib; copanlisib hydrochloride; crizotinib; cyclophosphamide; cytarabine; D-0502 (Inventisbio); dabrafenib mesylate; dacarbazine; dacomitinib; dactinomycin; daratumumab; daratumumab and hyaluronidase-fihj; darbepoetin alfa; darolutamide; dasatinib; daunorubicin hydrochloride; decitabine; defibrotide sodium; degarelix; denileukin diftitox; denosumab; dexamethasone; dexrazoxane hydrochloride; dinutuximab; docetaxel; doxorubicin hydrochloride; durvalumab; duvelisib; elastrant; elotuzumab; eltrombopag olamine; emapalumab-lzsg; enasidenib mesylate; encorafenib; enfortumab vedotin-ejfv; entrectinib; enzalutamide; epirubicin hydrochloride; epoetin alfa; erdafitinib; Eribulin mesylate; Erlotinib hydrochloride; Etoposide; Etoposide phosphate; Everolimus; Exemestane; Fam-trastuzumab deruxtecan-nxki; Fedratinib hydrochloride; Filgrastim; Fludarabine phosphate; Fluorouracil; Flutamide; Fostamatinib disodium; Fulvestrant; Gefitinib; Gemcitabine hydrochloride; Gemtuzumab ozogamicin; Gilteritinib fumarate; Glasdegib maleate; Glucarpidase; Goserelin acetate; Granisetron; Granisetron hydrochloride; Hydroxyurea; Ibritumomab tiuxetan; Ibrutinib; Idarubicin hydrochloride; Idealisib; Ifosfamide; Imatinib mesylate; Imiquimod; inotuzumab ozogamicin; recombinant interferon alfa-2b; iobenguane I-131; ipatasertib; ipilimumab; irinotecan hydrochloride; isatuximab-irfc; ivosidenib; ixabepilone; ixazomib citrate; lanreotide acetate; lapatinib ditosylate; larotrectinib sulfate; lasofoxifene; lenalidomide; lenvatinib mesylate; letrozole; leucovorin calcium; leuprolide acetate; lomustine; lorlatinib; LSZ102 (Novartis); lurbinectedine; LY3484356 (Lilly); megestrol acetate; melphalan; melphalan hydrochloride; mercaptopurine; methotrexate; midostaurin; Mitomycin; Mitoxantrone hydrochloride; Mogamulizumab-kpkc; Moxetumomab pasudotox-tdfk; Necitumumab; Nelarabine; Neratinib maleate; Nilotinib; Nilutamide; Niraparib tosylate monohydrate; Nivolumab; Obinutuzumab; Ofatumumab; Olaparib; Omacetaxine mepesuccinate; Ondansetron hydrochloride; Osimertinib mesylate; Oxaliplatin; Paclitaxel; Paclitaxel-albumin-stabilized nanoparticle formulation; Palifermin; Palonosetron hydrochloride; Pamidronate disodium; Panitumumab; Panobinostat; Pazopanib hydrochloride; Pegaspargase; Pegfilgrastim; Peginterferon alfa-2b; pembrolizumab; pemetrexed disodium; pemigatinib; pertuzumab; pexidartinib hydrochloride; plerixaphora; polatuzumab vedotina-piiq; pomalidomide; ponatinib hydrochloride; pralatrexate; prednisolone; procarbazine hydrochloride; propranolol hydrochloride; radium dichloride 223; raloxifene hydrochloride; ramucirumab; rasburicase; ravulizumab-cwvz; recombinant interferon alpha-2b; regorafenib; RG6171 (Roche); rhintodestrant; ripretinib; rituximab; rollapitant hydrochloride; romidepsin; romiplostim; rucapariba camzilata; ruxolitinib phosphate; Sacituzumab govitecan-hziy; SAR439859 (Sanofi); Selinexor; Selpercatinib; Selumetinib sulfate; SHR9549 (Jiansu Hengrui Medicine); Siltuximab; Sipuleucel-t; Sonidgiba; Sorafenib tosylate; Tagraxofuspa-erzs; Talazoparib tosylate; Talimogen laherparepvec; Tamoxifen citrate; Tazemetostat hydrobromide; Temozolomide; Temsirolimus; Thalidomide; Thioguanine; Thiotepa; Tisagenlecleicel; Tocilizumab; Topotecan hydrochloride; Toremifene; Trabectedin; Trametinib; Trastuzumab; Trastuzumab and hyaluronidase-oysk; trifluridine and tipiracil hydrochloride; tucatinib; uridine triacetate; valrubicin; vandetanib; vemurafenib; venetoclax; vinblastine sulfate; vincristine sulfate; vinorelbine tartrate; vismodegib; vorinostat; zanubrutinib; ziv-aflibercept; ZN-c5 (Zentalis); and zoledronic acid; or the free base, pharmaceutically acceptable salt, or solvate of any of the foregoing; or a combination thereof.

These and other aspects will become apparent from the ideas contained in this document.

PF-07104091 can be prepared as described in U.S. Patent No. 11,014,911.

PF-07220060 can be prepared as described in U.S. Patent No. 10,766,884.

EXAMPLES

The following examples are merely illustrative of the description and should not be construed as limiting the scope of the invention, since these examples and other equivalents thereof will become apparent to those skilled in the art in light of the present description and the appended claims.

Example 1. Inhibition of spheroid growth

Lung adenocarcinoma (LUAD) cell lines NCI-H1792 and NCI-H23 were obtained from the ATCC and maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum and penicillin-streptomycin according to ATCC recommendations. Cells were maintained in a humidified incubator at 37°C with 5% _CO2 .

Spheroid assays were performed in 96-well ultra-low attachment plates (ULA-96U) from Nexcelom & Thermo Fisher Scientific. Two hundred (200) NCI-H1792 or NCI-H23 cells were dispensed at 200 µl of complete growth medium per well (n=10-12 wells per treatment group) of each ultra-low attachment plate to ensure the formation of one spheroid per well with a diameter between 200 and 250 µm before treatment (the cell seeding amount was pre-optimized to ensure the formed spheroids had the desired size). To facilitate spheroid formation, dispensed cells were centrifuged at 220 g for 66 min in ultra-low attachment plates and allowed to form compact spheroids for 4 days before treatment. After spheroid formation, 150 μL of medium is aspirated from each well without disturbing the spheroid, and the same volume of fresh RPMI medium containing single agent compounds (palbociclib, PF-07220060, PF-07104091), or selected combinations thereof, is added. The final concentrations of each compound in the wells are: 30 nM palbociclib; 300 nM PF-07220060 or PF-07104091. DMSO (0.01%) is used as a vehicle control. DMSO and all compounds are diluted in cell medium. Medium and compounds are replenished twice weekly at 3- and 4-day intervals. Replenishment is accomplished by aspirating 150 μL of medium per well without disturbing the spheroid and then adding the same volume of premixed medium/compound solution to the spheroids. Spheroid diameters were quantified immediately after each medium change twice weekly (every 3rd or 4th day) throughout the assay.

Mean tumor spheroid diameters are plotted in GraphPad Prism 8, and AUC is calculated. Baseline AUC is defined as the mean tumor spheroid diameter on day 0 in vehicle control (DMSO). Spheroid growth inhibition, or SGI, is calculated as follows: SGI = (1 - AUC of treatment / AUC of DMSO) × 100%. SGI for all treatment groups is determined at the time point when vehicle-treated spheroids reach their maximum diameter (usually close to 1 mm, but may vary depending on the cell line); this corresponds to the last time point taken for vehicle-treated spheroids. SOM is calculated based on n = 10-12 wells per test group.

The growth of multicellular lung adenocarcinoma tumor spheroids (MCTS) is monitored over time to assess: (i) the magnitude of response (SGI) and (ii) the duration of response to the CDK4 inhibitor PF-07220060, both as a single agent and in combination with the CDK2 inhibitor PF-07104091.

PF-07220060 and PF-07104091 are expected to provide greater clinical tolerability (due to reduced dose-limiting toxicity) than palbociclib in humans (mean free plasma concentration of 30 nM). Therefore, spheroids were treated with 300 nM PF-07220060 or PF-07104091 and compared with palbociclib at a clinically relevant concentration of 30 nM. Treatment with PF-07104091 increased the sensitivity of NCT-H1792 and NCI-H23 spheroids to PF-07220060, indicating that the combination of both agents enhances the antitumor effect compared to either compound alone. Moreover, 300 nM PF-07220060 plus 300 nM PF-07104091 demonstrated superior inhibition of spheroid growth compared to 30 nM palbociclib plus 300 nM PF-07104091. The data are presented in Table 1.

Example 2. Inhibition of ER+ human breast cancer cell proliferation by PF-07104091 in combination with the CDK4 inhibitor PF-07220060

The combined effects of PF-07104091 and PF-07220060 were evaluated in a cell proliferation assay using MCF7 and T47D BC cell lines. Each compound demonstrated potent, dose-dependent growth inhibition in both cell models.

Drug combinations are analyzed using the dose equivalence principle and the Lowe volume estimate. When modeling synergy using the dose equivalence principle, the Lowe volume estimate for combination compounds should be higher than that of the self-crossover control, and the additivity index (CI) estimate for combination compounds should be lower than that of the self-crossover control. Combinations of PF-07104091 with PF-07220060 demonstrate favorable profiles for these two metrics, Lowe volume and CI, compared to the self-crossover control in the MCF7 breast cancer cell line (Table 2) and the T47D breast cancer cell line (Table 3). Isobolograms set to an anti-proliferation threshold of 70% confirm a synergistic response. Based on Lowe's volumes, CI values, and isobolograms, moderate combinational synergy is observed in these cell lines, with the highest combinatorial advantage observed in the MCF7 model.

Example 3. Antitumor efficacy of PF-07104091 in combination with PF-07220060 in breast cancer tumor models

Methods

In vivo efficacy assessment and statistical analysis

Female NSG mice (Jackson Lab) are implanted subcutaneously with a fragment (ranging in size from 27 ^mm3 to 64 ^mm3 ) in the dorsal region. All mice in the T47D, HCC1428, and ST941PBR studies are supplemented with 8.5 μg/mL estradiol-containing water (β-estradiol Sigma-Aldrich, Cat. #E2758-5G) and are given ad libitum until the end of the study. For in vivo efficacy studies, tumor volume and body weight are measured twice weekly. Tumor volume is calculated as [(Length × Width × Width)/2)]. TGI is calculated as 100*(1-ΔT/ΔC). ΔC(ΔT) is obtained by subtracting the mean tumor burden in the vehicle (treated) group on the first day of treatment (day 0) from the mean tumor burden in the vehicle (treated) group on the day of assessment. Statistical analysis was performed using ANCOVA when the mean tumor volume in vehicle-treated mice reached the tumor size cutoff.

In vivo efficacy evaluation of PF-07104091 in combination with PF-07220060 or palbociclib in a model of HR+/HER2- T47D breast cancer

The T47D model is established by implanting passage 3 tumor fragments into recipient mice. To establish T47D donor mice, tumor cells (5 x 10 ⁶ cells/mouse with 50% Cultrex® Basement Membrane Scaffold) are implanted subcutaneously into female NSG mice. Once reaching the 700-800 mm ³ range, the donor tumors are then transplanted into secondary recipient mice to expand the study. Once tumor volumes reach the 101 mm ³ -255 mm ³ range, tumor-bearing mice are randomly assigned to groups (n=8 per group) and administered with 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 150 mg/kg; 3) PF-07220060 60 mg/kg; 4) PD-0332991 10 mg/kg; 5) PF-07104091 150 mg/kg plus PF-07220060 60 mg/kg PF; 6) PF-07104091 150 mg/kg plus palbociclib 10 mg/kg. PF-07104091 (lot 016), PF-07220060 (lot 019), and PD-0332991 (lot GR08498) were administered po BID (7 hours apart). All mice were treated continuously for up to day 41. TGI was assessed at day 41 after the first dose.

Evaluation of the in vivo efficacy of PF-07104091 in combination with PF-07220060 or palbociclib in the HR+/HER2- HCC1428 breast cancer model.

The HCC1428 model is established by implanting passage 4 tumor fragments into recipient mice. To establish HCC1428 donor mice, tumor cells ( ^5x106 cells/mouse with 50% Cultrex® Basement Membrane Scaffold) are implanted subcutaneously into female NSG mice. After reaching the 700-800 ^mm3 range, the donor tumors are subsequently transplanted into secondary recipient mice to expand the study. When tumor volumes reach the 100 ^mm3 to 272 ^mm3 range, tumor-bearing mice are randomly assigned to groups (n=10 per group) and administered with 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 150 mg/kg; 3) PF-07220060 60 mg/kg; 4) PD-0332991 10 mg/kg; 5) PF-07104091 150 mg/kg plus PF-07220060 60 mg/kg PF; 6) PF-07104091 150 mg/kg plus palbociclib 10 mg/kg. PF-07104091 (lot 016), PF-07220060 (lot 019), and PD-0332991 (lot GR08498) were administered po BID (7 hours apart). All mice were treated continuously until day 42. TGI was assessed at day 42 after the first dose.

In vivo assay of palbociclib resistance, HR+/HER2- BC PDX model, ST941PBR

The ST941PBR palbociclib resistance model is obtained from XENOSTART™ LLC, San Antonio, Texas. A proprietary model is established by continuously treating ST941PBR tumor-bearing mice with PD-0332991 at 50 mg/kg p.o. q.d. plus fulvestrant at 10 mg/kg s.c. twice during the first week and then weekly thereafter. To expand donor mice for the TGI study, tumors are transplanted into recipient mice. One week after reimplantation, recipient mice receive continuous treatment with palbociclib plus fulvestrant (dosing regimen described above). After 1-2 serial in vivo expansions with the same treatment regimen, resistant tumors measuring 700 to 800 mm are reimplanted to expand the study. The remaining tumor fragments are frozen for future use.

To confirm treatment resistance, ST941PBR mice (p12) bearing tumors were treated with palbociclib (50 mg/kg) plus fulvestrant (10 mg/kg) using the same treatment regimen as described above.

In vivo efficacy evaluation of PF-07104091 in combination with PF-07220060 or palbociclib in the ST941PBR PDX model

Living donor mice are generated by implanting ST941PBR (p11) tumor fragments. Once tumor volumes reach the 700-800 ^mm3 range, donor tumors are then transplanted into secondary recipient mice to expand the study. To maintain resistant clones, ST941PBR tumor-bearing mice are treated with PD-0332991 at 50 mg/kg p.o., QD, plus fulvestrant at 10 mg/kg sc twice for the first week, then weekly for 10 weeks. Treatment begins one week after implantation until the day of randomization for study inclusion. Once tumor volumes reach the 121 ^mm3 to 22.8 ^mm3 range, tumor-bearing mice are randomly assigned to groups (n=10 per group) and administered 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 150 mg/kg; 3) PF-07220060 at 60 mg/kg; 4) PD-0332991 at 10 mg/kg; 5) PF-07104091 at 150 mg/kg plus PF-07220060 at 60 mg/kg PF; 6) PF-07104091 at 150 mg/kg plus palbociclib at 10 mg/kg; 7) PD-0332991 at 10 mg/kg plus fulvestrant at 10 mg/kg; 8) PD-0332991 50 mg/kg plus fulvestrant 10 mg/kg. PF-07104091 (lot 016), PF-07220060 (lot 019), PF-06873600 (lot 022), and PD-0332991 (lot GR08498) 10 mg/kg are administered po BID (7 hours apart); PD-0332991 (lot GR08498) 50 mg/kg is administered po QD and fulvestrant 10 mg/kg is administered sc twice during the first week and then weekly. All mice are treated continuously until day 28. TGI is assessed at day 28 after the first dose.

Results and discussions

PF-07104091 is also being evaluated as a single agent and in combination with palbociclib or PF-07220060 in three models of HR+, HER2- BC: T47D, HCC1428, and palbociclib-resistant ST941PBR PDX. Treatment is administered po, BID, at a mg/kg (mg/kg) dose as shown in Table 4 (except for the palbociclib 50 mg/kg QD and fulvestrant SC arm twice in the first week and then weekly).

No significant changes in body weight or other clinical observations were observed throughout the treatment period in any of the 3 models in 10 subjects (clinical pathology was not performed).

In the T47D model, single-agent treatment with PF-07104091, PF-07220060, and palbociclib demonstrated significant TGI efficacy compared to vehicle (p<0.05). Treatment with PF-07104091 plus PF-07220060 demonstrated significantly increased efficacy (TGI 106%) compared to either PF-07220060 (TGI 73%) or PF-07104091 (TGI 82%) monotherapy (p<0.05 vs. 15 for each of the individual agents). PF-07104091 in combination with palbociclib (TGI 100%) also demonstrated significantly increased efficacy compared to any monotherapy (p<0.05 compared to any of the individual agents).

In the HCC1428 model, treatment with each individual agent PF-07104091, PF-07220060, and palbociclib also demonstrated significant TGI efficacy compared with vehicle (p<0.05). Treatment with PF-07104091 plus PF-07220060 demonstrated significantly increased efficacy (TGI 110%) compared with PF-07220060 monotherapy (TGI 89%) or PF-07104091 (TGI 95%) (Table 4).

In the palbociclib-resistant ST941PBR PDX model, treatment with palbociclib (10 mg/kg BID or 50 mg/kg QD) in combination with fulvestrant demonstrated a significant but minimal response (30% and 26% TGI, respectively, compared with vehicle), confirming acquired resistance (Table 4). PF-07104091 in combination with palbociclib resulted in a significant increase in TGI (49%) compared with vehicle or PF-07104091 monotherapy (34% TGI) or palbociclib. Treatment with PF-07104091 in combination with PF-07220060 significantly improves efficacy compared to either monotherapy (TGI for the combination 98% versus 34% for PF-07104091 monotherapy and 28% for PF-07220060 monotherapy). The combination of PF-07104091 plus PF-07220060 results in a significant improvement in efficacy compared to the combination of PF-07104091 plus palbociclib in this palbociclib-resistant ST 941PBRPDX model.

Example 4. Antitumor efficacy of low dose PF-07104091 in combination with PF-07220060 in breast cancer tumor models

Methods

In vivo efficacy assessment and statistical analysis

Female NSG mice (Jackson Lab) receive a subcutaneous implantation of a fragment (ranging in size from 27 ^mm3 to 64 ^mm3 ) in the dorsal region. All mice in the HCC1428, MCF7, and ST941PBR studies are supplemented with 8.5 μg/mL estradiol-containing water (β-estradiol Sigma-Aldrich, Cat. #E2758-5G) and are given ad libitum until the end of the study. For in vivo efficacy studies, tumor volume and body weight are measured twice weekly. Tumor volume is calculated as [(Length × Width × Width)/2)]. TGI is calculated as 100*(1-ΔT/ΔC). ΔC(ΔT) is obtained by subtracting the mean tumor burden in the vehicle (treated) group on the first day of treatment (day 0) from the mean tumor burden in the vehicle (treated) group on the day of assessment. Statistical analysis was performed using ANCOVA when the mean tumor volume in vehicle-treated mice reached the tumor size cutoff.

Evaluation of the in vivo efficacy of PF-07104091 in combination with PF-07220060 or palbociclib in the HR+/HER2- HCC1428 breast cancer model.

The HCC1428 model is generated by implanting passage 1 tumor fragments into recipient mice. To establish HCC1428 donor mice, tumor cells ( ^5x106 cells/mouse with 50% Cultrex® Basement Membrane Scaffold) are implanted subcutaneously into female NSG mice. When they reach the 700-800 ^mm3 range, the donor tumors are then transplanted into secondary recipient mice to expand the study. When tumor volumes reach the 119 ^mm3 to 267 ^mm3 range, tumor-bearing mice are randomly assigned to groups (n=10 per group) and administered with 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 120 mg/kg; 3) PF-07220060 at a dose of 60 mg/kg; 4) PF-06873600 at a dose of 30 mg/kg; 5) Palbociclib at a dose of 10 mg/kg plus fulvestrant at a dose of 10 mg/kg; 6) PF-07104091 at a dose of 120 mg/kg plus PF-07220060 at a dose of 20 mg/kg; 7) PF-07104091 at a dose of 75 mg/kg plus PF-07220060 at a dose of 40 mg/kg; 8) PF-07104091 at a dose of 35 mg/kg plus PF-07220060 at a dose of 60 mg/kg. PF-07104091 (lot 022), PF-07220060 (lot CPo126812-01-SY-5700-01), PF-06873600 (lot 022), and PD-0332991 (lot GR08498) were administered (po) BID (7 hours apart), while fulvestrant (lot 20160224-2) at a dose of 10 mg/kg was administered subcutaneously twice in the first week and then weekly. All mice received treatment continuously until day 42. TGI was assessed at day 42 after the first dose.

In vivo efficacy evaluation of PF-07104091 in combination with PF-07220060 or palbociclib in the HR+/HER2- MCF7 breast cancer model.

The MCF7 model is established by implanting donor tumor fragments into recipient mice. To establish MCF7 donor mice, tumor cells (5 x 10 ⁶ cells/mouse with 50% Cultrex® Basement Membrane Matrix) are implanted subcutaneously into female NSG mice. When they reach the 700-800 mm ³ range, the donor tumors are then transplanted into secondary recipient mice to expand the study. When the tumor volume reaches the 141 mm ³ to 231 mm ³ range, tumor-bearing mice are randomly assigned to groups (n=10 per group) and administered with 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 120 mg/kg; 3) PF-07220060 at a dose of 60 mg/kg; 4) PF-06873600 at a dose of 30 mg/kg; 5) Palbociclib at a dose of 10 mg/kg plus fulvestrant at a dose of 10 mg/kg; 6) PF-07104091 at a dose of 120 mg/kg plus PF-07220060 at a dose of 20 mg/kg; 7) PF-07104091 at a dose of 75 mg/kg plus PF-07220060 at a dose of 40 mg/kg; 8) PF-07104091 at 35 mg/kg plus PF-07220060 at 60 mg/kg. PF-07104091 (lot 022), PF-07220060 (lot CPo126812-01-SY-5700-01), PF-06873600 (lot 022), and PD-0332991 (lot GR08498) are administered (po) BID (7 hours apart), while fulvestrant (lot 20160224-2) at 10 mg/kg is administered sc twice during the first week and then weekly. All mice receive treatment continuously until day 29. TGI is assessed on day 29 after the first dose.

In vivo establishment of acquired resistance to palbociclib in the HR+/HER2- BC PDX, ST941PBR model

The ST941PBR palbociclib resistance model is obtained from XENOSTART™ LLC, San Antonio, Texas. A proprietary model is generated by continuously treating ST941PBR tumor-bearing mice with PD-0332991 at 50 mg/kg p.o. QD plus fulvestrant at 10 mg/kg s.c. twice during the first week and then weekly thereafter. To generate donors for the TGI study, tumors are transplanted into recipient mice. One week after reimplantation, recipient mice receive continuous treatment with palbociclib plus fulvestrant (dosing regimen described above). After 1-2 serial in vivo expansions with the same treatment regimen, resistant tumors measuring 700 to 800 ^mm3 are reimplanted to expand the study. The remaining tumor fragments are frozen for future use.

To confirm treatment resistance, ST941PBR mice (p14) bearing tumors were treated with palbociclib (50 mg/kg) plus fulvestrant (10 mg/kg) using the same treatment regimen as described above.

In vivo Evaluation of PF-07104091 in Combination with PF-07220060 or Palbociclib in the ST941PBR PDX Model Live donor mice are generated by implanting ST941PBR (p13) tumor fragments. After reaching the 700-800 ^mm3 range, donor tumors are then transplanted into secondary recipient mice to expand the study. To maintain resistant clones, ST941PBR tumor-bearing mice are treated with PD-0332991 at 50 mg/kg PO, QD, plus fulvestrant at 10 mg/kg SC twice for the first week, then weekly for 10 weeks. Treatment begins one week after implantation until the day of randomization for study inclusion. When tumor volumes reached a range of 137 ^mm3 to 216 ^mm3 , tumor-bearing mice were randomly assigned to groups (n=10 per group) and administered with 1) vehicle (0.5% MC with 0.1% Tween 80 in water); 2) PF-07104091 at 120 mg/kg; 3) PF-07220060 at 60 mg/kg; 4) PF-06873600 at 30 mg/kg; 5) Palbociclib at 10 mg/kg plus fulvestrant at 10 mg/kg; 6) PF-07104091 at 120 mg/kg plus PF-07220060 at 20 mg/kg; 7) PF-07104091 at a dose of 75 mg/kg plus PF-07220060 at a dose of 40 mg/kg; 8) PF-07104091 at 35 mg/kg plus PF-07220060 at 60 mg/kg. PF-07104091 (lot 021), PF-07220060 (lot CPol26812-01-SY-5700-01), PF-06873600 (lot 022), and PD-0332991 (lot GR08498) are administered (po) BID (7 hours apart), while fulvestrant (lot 20160224-2) is administered at 10 mg/kg sc twice during the first week and then weekly. All mice receive treatment continuously until day 28. TGI is assessed at day 28 after the first dose.

Results and discussions

Lower dose levels of PF-07104091+PF-07220060 are also being evaluated to test whether the combination can improve efficacy compared with monotherapy, PF-06873600, and SOC palbociclib + fulvestrant in three HR+, HER2-BC models: MCF7, HCC1428, and palbociclib-resistant ST941PBR PDX. Treatment is administered po, BID, at the mg/kg (mcg) dose listed in Table 5 (except fulvestrant at 10 mg/kg s.c. twice in the first week and weekly thereafter). No significant changes in body weight or other clinical observations were observed throughout the treatment period in any of the three models (clinical pathology is not performed).

In the MCF7 model, SOC palbociclib 10 mg/kg + fulvestrant 10 mg/kg and PF-07104091 120 mg/kg, PF-07220060 60 mg/kg, PF-06873600 30 mg/kg as single agent treatments demonstrated significant TGI efficacy compared to vehicle (TGI 84%, 60%, 79%, and 70%, respectively; p<0.05). Low dose treatment groups PF-07104091 + PF-07220060 - PF-4091 35 mg/kg + PF-0060 60 mg/kg, PF4091 75 mg/kg + PF-0060 40 mg/kg, and PF-4091 120 mg/kg + PF0060 20 mg/kg, demonstrated significant regression (TGI 106%, 106%, 105%) compared to vehicle, SOC palbociclib 10 mg/kg + fulvestrant 10 mg/kg, and monotherapy with PF-07104091 at a dose of 120 mg/kg, PF-07220060 at 60 mg/kg, and PF-06873600 at 30 mg/kg. There was also no significant difference in efficacy between the low-dose PF-4091+PF-0060 groups (p>0.05) in the MCF7 model.

In the HCC1428 model, SOC palbociclib 10 mg/kg + fulvestrant 10 mg/kg and PF-07104091 at a dose of 120 mg/kg, PF-07220060 at a dose of 60 mg/kg, PF-06873600 at a dose of 30 mg/kg when treated with a single agent demonstrate significant TGI efficacy compared to vehicle (TGI 70%, 79%, 70% and 97%, respectively; p<0.05). The low dose treatment groups PF-07104091+PF-07220060 - PF-4091 35 mg/kg + PF-0060 60 mg/kg, PF4091 75 mg/kg + PF-0060 40 mg/kg, and PF-4091 120 mg/kg + PF0060 20 mg/kg - demonstrated significant regression (TGI 110%, 110%, 109%, respectively) compared to vehicle, S0C palbociclib 10 mg/kg + fulvestrant 10 mg/kg, and PF-07104091 120 mg/kg, PF-07220060 60 mg/kg, and PF-06873600 30 mg/kg monotherapy. There was also no significant difference in efficacy between the low-dose PF-4091+PF-0060 groups (p>0.05) in the HCC1428 model.

In palbociclib-resistant ST941PBR PDX models, treatment with palbociclib (10 mg/kg BID) in combination with fulvestrant demonstrated a minimal response (14% TGI compared with vehicle), confirming acquired resistance (Table 5). PF-07220060 at 60 mg/kg demonstrated a minimal response (TGI 19%, p>0.05), and PF-07104091 at 120 mg/kg and PF-0873600 at 30 mg/kg demonstrated significant but modest efficacy compared with vehicle (TGI 36% and 47%, respectively; p<0.05). Low doses of the PF-07104091+PF-07220060 treatment groups - PF-4091 35 mg/kg + PF-0060 60 mg/kg, PF409175 mg/kg + PF-0060 40 mg/kg, and PF-4091 120 mg/kg + PF-0060 20 mg/kg - demonstrated significant inhibition (TGI 87%, 98%, 94%, respectively) compared to vehicle, SOC palbociclib 10 mg/kg + fulvestrant 10 mg/kg, and monotherapy with PF-07104091 at 120 mg/kg, PF-07220060 at 60 mg/kg, and PF-06873600 at 30 mg/kg. Among the low dose groups, there was no significant difference in efficacy between PF-4091 75 mg/kg + PF-0060 40 mg/kg and PF-4091 120 mg/kg + PF-0060 20 mg/kg, but significantly different between PF-4091 75 mg/kg + PF-0060 40 mg/kg and PF-4091 120 mg/kg + PF-0060 20 mg/kg versus PF-4091 35 mg/kg + PF-0060 60 mg/kg (p<0.05).

The results demonstrate the ability to reduce the impact of either agent or both and maintain maximum tumor control (regression in the study) in primary disease cell line models (MCF7 and HCC1428). In the STR941-PBR treatment-resistant model, reducing the CDK2i dose in the combination regimen to 35 mg BID does differ from other combination groups tested, demonstrating a slightly lower TGI in the study and more robust recovery after treatment.

Example 5. Combinatorial activity of PF-07104091 and PF-07220060 in lung adenocarcinomas

The combinatorial activity of PF-07104091 and PF-07220060 was evaluated in LUAD both as single agents (single agents) and in combination. The data are presented in Table 6.

Example 6. Preliminary assessment of the risk of drug-drug interactions (DDI)

A preliminary assessment of the substrate/prey DDI risk for PF-07104091 was performed using a static mechanistic model with PF-07220060 at clinically relevant concentrations. (Fahmi OA, Hurst S, Plowchalk D, et al. Comparison of different algorithms for predicting clinical drug-drug interactions based on the use of CYP3A4 in vitro data: predictions of 10 compounds as precipitants of interaction. Drug Metab Dispos 2009; 37(8):1658–66.)

Based on potential combination doses (300 mg PF-07220060 and 75 mg PF-07104091), PF-07220060 is predicted to exhibit time-dependent inhibition (TDI) of cytochrome P450 3A (CYP3A) based on the potential interfering effect of PF-07104091 in combination, resulting in an AUC ratio (AUCR) of ~2.

Apart from the potential CYP3A DDI, there are no additional data to indicate a significant risk of DDI between PF-07104 091 and PF-07220060 as either offender or substrate/prey at clinically relevant concentrations, nor when letrozole or fulvestrant are added to this combination.

Example 7. An open-label, multicenter, non-randomized, dose-escalation, expansion study of Phase 1b/2 to evaluate the safety, tolerability, PK, PD, and antitumor activity of PF-07220060 administered in combination with PF-07104091.

An open-label, multicenter, non-randomized, dose-escalation/expansion study to evaluate the safety, tolerability, PK, PD, and antitumor activity of PF-07220060 and PF-07104091 (double combination) in participants with metastatic breast cancer (mBC) or other advanced solid tumors (Part 1), and when administered with endocrine therapy (ET) in participants with metastatic/advanced mBC (Part 2). At the data cutoff, 12 participants are receiving double combination therapy in Part 1 (dose escalation) across 4 dose cohorts.

In Part 2, approximately 6 participants will be enrolled for each of the two triple combinations (PF-07220060, PF-07104091, and fulvestrant or PF-07220060, PF-07104091, and letrozole) to establish the safety and tolerability of the combinations. Part 2A will include participants previously treated with a CDK4/6 inhibitor and will receive the triple combination of PF-07220060, PF-07104091, and fulvestrant. Part 2B will include participants previously untreated with CDK4/6 inhibitors but previously treated with 1ET and will receive the triple combination of PF-07220060, PF-07104091, and fulvestrant. Part 2C will enroll participants previously untreated with CDK4/6 and ET inhibitors for advanced disease and will receive the triple combination of PF07220060, PF-07104091, and letrozole.

Dose escalation of PF-07220060 and PF-07104091 will continue in a stepwise fashion according to the recommendations of the Bayesian logistic regression model (BLRM). Dose escalation will continue until the MTD of the combination of PF-07220060 and PF-07104091 is determined or the criteria for stopping further escalation are met. The highest dose of PF-07220060 and PF-07104091 tested in this study will not be higher than the MTD for monotherapy/the recommended dose for expansion (RDE) of each drug, as determined in the single-agent studies, respectively. Dose escalation decisions will also take into account all available clinical data from this study.

To assess PK and DDI potential, spiked PK samples will be collected for both PF-07220060 and PF-07104091. To assess potential DDI, Part 1 participants will receive a single dose of PF-07104091 on day -1 with PK assessments within 24 hours of the PF-07104091 dose. Initially, participants will receive both PF-07220060 and PF-07104091 on day 1 of cycle 1 in a continuous schedule specified as a BID schedule, and a multiple-dose PK assessment will be performed on day 15 of cycle 1. PK assessments may be modified (e.g., discontinuing the day -1 assessment of cycle 1) based on evolving data from the initial DDI assessment cohorts.

Part 1 will include participants with metastatic/advanced breast cancer and other advanced or metastatic solid tumors without a standard alternative treatment option to determine the dual MTD/RDE. Each cohort will include approximately 2-4 participants. At least 6 participants will receive the recommended combination doses of PF-07220060 and PF-07104091 before dose expansion.

Participants with VS may also receive fulvestrant after at least 2 cycles of the combination of PF-07220060 and PF-07104091 at the discretion of the investigator and with sponsor approval if:

The current dose of PF-07220060 and PF-07104091 received by participants as combination therapy was determined to be safe by the BLRM;

• the participant did not experience an adverse event (AE) that met the DLT criteria, or;

• The DLT-eligible AE resolved and the participant resumed combination therapy with PF-07220060 and PF-07104091 for at least 28 days without another DLT-eligible AE.

Part 2 (dose expansion) will enroll up to 3 cohorts of participants with advanced or metastatic HR-positive HER2-negative BC to treatment with the triple combination of PF-07220060, PF07104091, and ET (letrozole or fulvestrant).

• Part 2A will include participants previously treated with CDK4/6 inhibitors.

• Part 2B will include participants who have not been previously treated with CDK4/6 inhibitors but have been previously treated with 1ET.

• Part 2C will include participants who have not previously been treated with CDK4/6 and ET inhibitors for advanced disease.

Participants in Parts 2A and 2B will receive the triple combination of PF-07220060, PF-07104091, and fulvestrant starting in Cycle 1, and participants in Part 2C will receive the triple combination of PF07220060, PF-07104091, and letrozole starting in Cycle 1.

To select the RDE combination, a dose escalation or reduction approach will be used at the full or intermediate dose of PF-07220060 or PF-07104091, depending on emerging clinical data. The BLRM, along with the EWOC criteria, will be used in Part 1 to define MTB.

All cycles are 28 days long, and treatment will continue until disease progression, uncontrolled toxicity, the participant or investigator decides to discontinue treatment, or the study is terminated. Participants experiencing toxicity, including DLT, may be treated with dose modification or discontinuation of either PF-07220060, PF-07104091, or both.

PF-07220060 and PF-07104091 will be administered orally BID as a continuous daily dose. To ensure optimal dosing of PF-07220060 and PF-07104091 in combination with endocrine therapy, i.e., letrozole or fulvestrant, alternative dosing regimens, schedules, and PK time points may be considered based on emerging PK data, safety, tolerability, laboratory data, and PD.

As of the data cutoff date, a total of 12 participants are receiving PF-07220060 in combination with PF-07104091 (a selective CDK2 inhibitor) in Part 1 (dose escalation) across 4 dose cohorts.

Demography

Overall, a total of 12 participants (11 women and 1 man) with a mean age of 55.25 years (SD: 11.19) are receiving PF-07220060+PF-07104 091. Participants have advanced cancers including HR+HER2- breast cancer (n=8), small cell carcinoma (n=1), ovarian cancer (n=1), small cell lung cancer (n=1), and not yet reported (n=1).

Adverse events of any cause arising during treatment

Of the 12 participants receiving treatment, 10 (83.3%) participants reported a total of 88 treatment-emergent adverse events (TEAEs).

The most frequently reported (≥20%) events of any cause were nausea (58.3%), diarrhea, decreased neutrophil and white blood cell counts (50.0% each), anemia (41.7%), decreased lymphocyte count (33.3%), fatigue, decreased platelet count, and vomiting (25.0% each).

No grade 4 TEAEs were reported. One grade 5 TEAE of disease progression was reported during the study.

Treatment-related adverse events caused by treatment

A total of 10 (83.3%) of 12 participants reported treatment-related TEAEs.

The most commonly reported (≥20%) treatment-related AEs were nausea (58.3%), diarrhea, neutrophil and white blood cell count decreased (50.0% each), lymphocyte count decreased (33.3%), anemia, fatigue, platelet count decreased, and vomiting (25.0% each).

No treatment-related grade 4 or 5 TEAEs were reported during the study.

Dose-limiting toxicity

At the data cutoff date, one participant receiving PF-07220060 300 mg BID+PF-07104091 150 mg BID experienced one DLT (grade 3 fatigue).

Side effects leading to discontinuation

None of the participants discontinued the study drug due to adverse events.

Serious adverse events and death

No serious adverse events (SAEs) or deaths related to study treatment were reported. One participant in the Part 1 PF-07220060 200 mg BID + PF-07104091 75 mg BID treatment group reported an SAE of shortness of breath. Another participant in the Part 1 PF-07220060 300 mg BID + PF-07104091 150 mg BID treatment group reported an SAE of disease progression leading to death. Both SAEs were unrelated to study treatment.

Claims (45)

1. A method of treating cancer in a subject in need thereof, comprising administering to the subject: (a) amount of compound (A): (PF-07104091) or its hydrate, and (b) amount of compound (B): (PF-07220060) or its hydrate, wherein amounts (a) and (b) together are effective in treating cancer and wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer and stomach cancer. 2. The method of claim 1, further comprising administering to the subject: (c) an amount of an additional anti-cancer agent; wherein the amounts (a), (b), and (c) together are effective in treating cancer. 3. The method according to claim 1 or 2, wherein the cancer is hormone receptor positive (HR+) breast cancer and the additional anticancer agent is an endocrine therapeutic agent selected from the group consisting of an aromatase inhibitor, a selective estrogen receptor modulator (SERM) and a selective estrogen receptor biodegrader (SERD). 4. The method of claim 3, wherein the endocrine therapeutic agent is letrozole or fulvestrant. 5. The method according to any one of paragraphs 1-4, wherein the compound (A) is a monohydrate (PF-07104091). 6. The method of claim 5, wherein the amount of PF-07104091 is from 75 mg to 500 mg BID. 7. The method of claim 6, wherein the amount of PF-07104091 is 75 mg BID, 150 mg BID, or 225 mg BID. 8. The method according to any one of paragraphs 1-7, wherein the compound (B) is a hydrate (PF-07220060). 9. The method of claim 8, wherein the amount of PF-07220060 is from 100 mg to 500 mg BID. 10. The method of claim 9, wherein the amount of PF-07220060 is 100 mg BID, 200 mg BID, or 300 mg BID. 11. The method according to any one of claims 1-10, wherein the cancer is a cancer resistant to palbociclib. 12. The method according to any one of claims 1-10, wherein the subject has previously been treated with a CDK4/6 inhibitor. 13. The method of claim 12, wherein the CDK4/6 inhibitor is palbociclib. 14. A combination for the treatment of cancer comprising: (a) compound (A): (PF-07104091) or its hydrate; and (b) compound (B): (PF-07220060) or its hydrate, wherein the cancer is selected from the group consisting of breast cancer, lung cancer, ovarian cancer, peritoneal cancer, fallopian tube cancer, bladder cancer, colon cancer, uterine cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, and stomach cancer. 15. The combination of claim 14, further comprising (c) an additional anticancer agent; wherein the combinations (a), (b) and (c) are effective in treating cancer. 16. The combination of claim 14 or 15, wherein the cancer is hormone receptor positive (HR+) breast cancer and the additional anticancer agent is an endocrine therapeutic agent selected from the group consisting of an aromatase inhibitor, a selective estrogen receptor modulator (SERM), and a selective estrogen receptor biodegrader (SERD). 17. The combination of claim 16, wherein the endocrine therapeutic agent is letrozole or fulvestrant. 18. The combination according to any one of paragraphs 14-17, wherein the compound (A) is a monohydrate (PF-07104091). 19. The combination according to any one of paragraphs 14-18, wherein the compound (B) is a hydrate (PF-07220060).