Combination therapies using CDK4 inhibitors with PD-1 axis binding antagonists

Abstract

This invention relates to, in part, a method for treating cancer by administering a CDK4 inhibitor in combination with a PD-1 axis binding antagonist, and optionally an CDK2 inhibitor to a subject in need thereof.

Description

[0001] PC073220A

[0002] COMBINATION THERAPIES USING CDK4 INHIBITORS WITH PD- AXIS BINDING ANTAGONISTS FIELD

[0003] The present invention relates to combination therapies useful for the treatment of cancers. In particular, the invention relates to combination therapies which comprise administering a CDK4 inhibitor (optionally with a CDK2 inhibitor) in combination with a PD-1 axis binding antagonist. The invention also relates to associated methods of treatment, pharmaceutical compositions, and pharmaceutical uses. The methods and compositions are useful for any indication for which the therapeutic is itself useful in the detection, treatment and / or prevention of a disease, disorder, or other condition of a subject.

[0004] BACKGROUND

[0005] Cyclin-dependent kinases (CDKs) and related serine / threonine protein kinases are vital enzymes that regulate cell division and proliferation. CDK4 / 6 inhibitors, including palbociclib, ribociclib, and abemaciclib, have become standard treatments for patients with hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative (HR+ / HER2-) advanced or metastatic breast cancer when combined with endocrine therapy (Morrison, L., et al., Nature Reviews Clinical Oncology, 2024, 21, 89-105). However, these inhibitors often cause dose-limiting hematologic toxicities, foremost neutropenia.

[0006] Atirmociclib, a CDK4 selective inhibitor currently under clinical investigation, offers a more selective inhibition profile, potentially reducing the risk of neutropenia and other side effects associated with CDK6 inhibition. Despite advancements in CDK4 inhibitor development, there remains a significant need for improved combination therapies for cancer treatment.

[0007] The programmed death 1 (PD-1) receptor and PD-1 ligands 1 and 2 (PD-L1 and PD-L2, respectively) play integral roles in immune regulation, PD-1 is expressed by activated T cells, B cells, and myeloid cells. Notably, the majority of tumor infiltrating T lymphocytes overexpress PD- 1 relative to T lymphocytes in normal tissues and peripheral blood T lymphocytes (Ahmadzadeh et al., Blood, 2009 114(8):1537). PD-1 is activated by its ligands, PD-L1 and PD-L2, which are expressed by stromal cells, tumor cells, or both, this interaction initiates T-cell death and localized immune suppression (Dong et al., Nat Med 1999; 5:1365-69; Freeman et al., J Exp Med 2000; 192:1027-34), potentially providing an immune-tolerant environment for tumor development and growth. Conversely, inhibition of this interaction can enhance local T-cell responses and mediate antitumor activity in nonclinical animal models (Iwai Y, et al., Proc Natl Acad Sci USA 2002; 99:12293-97).

[0008] PD-L1 is a cell-surface protein and member of the B7 family, is found on almost all types of lymphohematopoietic cells and is expressed at low levels by resting T cells, B cells, macrophages and dendritic cells and is further up regulated by an anti-CD40 antibody for B cells, anti-CD3 antibody for T cells, anti-CD40 antibody, IFNy and granulocyte macrophage colony¬ stimulating factor (GM-CSF) for macrophages and / or anti-CD40 antibody, IFNy, IL-4, IL-12 and GM-CSF for Dendritic cells (DCs). PD-L1 is also expressed by some non-hemoatopoietic cells and is overexpressed in many cancers, wherein its overexpression is often associated with poor prognosis (Okazaki T et al., Intern. Immun. 2007 19(7):813) (Thompson R H et al., Cancer Res 2006, 66(7):3381). The majority of tumor infiltrating T lymphocytes predominantly express PD-1, in contrast to T lymphocytes in normal tissues and peripheral blood. PD-1 on tumor-reactive T cells can contribute to impaired antitumor immune responses (Ahmadzadeh et al., Blood 2009 1 14(8): 1537). This may be due to exploitation of PD-L1 signaling mediated by PD-L1 expressing tumor cells interacting with PD-1 expressing T cells to result in attenuation of T cell activation and evasion of immune surveillance (Sharpe et al., Nat Rev 2002) (Keir ME et al., Annu. Rev. Immunol. 2008, 26:677). Therefore, inhibition of the PD-L1 / PD-1 interaction may enhance CD8+ T cell-mediated killing of tumors.

[0009] The other known ligand for PD-1, PD-L2, is a cell surface protein expressed by antigen presenting cells, including dendritic cells, with expression also found in other non-hematopoietic tissues. Inhibition of PD-1 axis signaling through its direct ligands (e.g., PD-L1, PD-L2) has been proposed as a means to enhance T cell immunity for the treatment of cancer (e.g., tumor immunity). Similar enhancements to T cell immunity have been observed by inhibiting the binding of PD-L1 to the binding partner B7-1 (Ribas A. et a!., Science, 2018, 359: 1350-1355).

[0010] The present inventors have discovered that combination therapy with a CDK4 inhibitor and a PD-1 axis binding antagonist results in a synergistic inhibition of tumor growth compared to monotherapy with either agent alone. The combinations, methods and uses of the present invention are believed to offer one or more advantages, such as greater efficacy than treatment with either therapeutic agent alone, potential to reduce drug-drug interactions, potential to enable an improved dosing schedule, potential to reduce side effects, potential to overcome resistance mechanisms.

[0011] SUMMARY OF THE INVENTION

[0012] This invention relates to therapeutic methods, combinations, and pharmaceutical compositions for use in the treatment of cancer. Disclosed herein include methods of using a combination therapy that comprises a CDK4 inhibitor in combination with a PD-1 axis binding antagonist to treat cancer in a subject.

[0013] According to a first embodiment of the invention, there Is provided a method of treating cancer in a subject in need thereof comprising administering to a subject

[0014] (a) an effective amount of a CDK4 inhibitor having a Formula (I):

[0015]

[0016] wherein:

[0017] R1is H, F or Cl;

[0018] R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by Rs;

[0019] R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by Rs:

[0020] R is H or F; and

[0021] each R5and R6is independently OH, F or Ci-C2alkoxy; and

[0022] (b) an effective amount of a PD-1 axis binding antagonist; and

[0023] (c) optionally an effective amount of a CDK2 inhibitor.

[0024] According to a second embodiment of the invention, there is provided a combination comprising:

[0025] (i) a CDK4 inhibitor having a Formula (I):

[0026]

[0027] wherein:

[0028] R1is H, F or Cl;

[0029] R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R5;

[0030] R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionaily substituted by Rs:

[0031] R4is H or F; and

[0032] each R5and R6is independently OH, F orCi-C2alkoxy;

[0033] (ii) a PD-1 axis binding antagonist; and

[0034] (iii) optionally tegtociciib (a CDK2 inhibitor);

[0035] for use in the treatment of cancer in a subject.

[0036] According to a third embodiment of the invention, there is provided a combination comprising:

[0037] a. (i) atirmociclib; and (ii) a PD-1 axis binding antagonist; or b. (i) atirmociclib: (ii) PD-1 axis binding antagonist; and (iii) tegtociclib;

[0038] for use in the treatment of cancer in a subject.

[0039] According to a fourth embodiment of the invention, there are provided kits comprising one or more of the compositions of the invention.

[0040] Described below are embodiments of the invention, where for convenience Embodiment 1 (E1) is identical to the method of selecting a subject having a cancer provided above, and Embodiment 2 (E2) is identical to the method of treating a cancer in a subject provided above.

[0041] it is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.

[0042] BRIEF DESCRIPTION OF THE DRAWINGS FIG, 1 qRT-PCR of antigen (AG) presentation and interferon (IFN) signaling genes from human HR+ HER2- breast cancer, including MCF7, palbociclib resistant MCF7 (MCF7PR), T47D, and HCC1428, after 8-day treatment with vehicle (Veh), 30 nM palbociclib, 1 pM atirmociclib (At), 1 pM CDK2 inhibitor tegtociclib (PF-07104091) (CDK2I), or the combination (At + CDK2I). Heatmaps represent the average gene expression (0=2) relative to vehicle control, normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH).

[0043] FIG. 2 qRT-PCR of antigen (AG) presentation and interferon (IFN) signaling genes from human NSCLC cells lines, including H1792, H1573, HCC2108, HCC78 and H2087, after 8-day treatment with vehicle (Veh), 30 nM palbociclib, 1 pM atirmociclib (At), 1 pM CDK2 inhibitor tegtociclib (PF-07104091) (CDK2I), or the combination (At + CDK2I). Heatmaps represent average gene expression (n=2) relative to vehicle control, normalized to GAPDH.

[0044] FIG. 3 PDL1 / HLA-ABC flow cytometry plots from HCC78 cells after 8-day treatment with vehicle (Veh), 30 nM palbociclib, 1 pM atirmociclib (At), 1 pM CDK2 inhibitor tegtociclib (PF- 07104091) (CDK2I), or the combination (At +CDK2I). PDL1+ / HLA-ABC+ cells were quantified in the enclosed upper right square area (%).

[0045] FIG. 4 Summary of fold change in median fluorescence intensity (MFI) of PDL1 or HLA- ABC signals, from data in FIG. 3.

[0046] FIG. 5 / n vivo tumor growth in MC38R syngeneic mouse models. Mice were treated with vehicle (Veh), 10 mg / kg BID palbociclib (Pa), 10 mg / kg Q3Dx3, Q10D PD1 antibody PF- 06937004 (aPD1), or combinations thereof. Each data point is the mean value from 12 mice (±SEM). Percent TGI for treatment arms is shown. Asterisk indicates independently derived proliferation IC50 (pM) for atirmociclib in MC38R cell lines in vitro. FIG. 6 In vivo tumor growth in MC38R syngeneic mouse models. Mice were treated with vehicle (Veh), 60 mg / kg BID atirmociclib (At), 10 mg / kg Q3Dx3, Q10D PD1 antibody PF- 06937004 (aPD1), or combinations thereof. Each data point is the mean value from 12 mice (±SEM). Percent TGI for treatment arms is shown. Asterisk indicates independently derived proliferation IC50 (pM) for atirmociclib in MC38R cell lines in vitro.

[0047] FIG. 7 In vivo tumor growth in CT26 syngeneic mouse models. Mice were treated with vehicle (Veh), 10 mg / kg BID palbociclib (Pa), 10 mg / kg Q3Dx3, Q10D PD1 antibody PF- 06937004 (aPD1), or combinations thereof. Each data point is the mean value from 12 mice (±SEM). Percent TGI for treatment arms is shown. Asterisk indicates independently derived proliferation IC50 (pM) for atirmociclib in CT26 cell lines in vitro.

[0048] FIG. 8 In vivo tumor growth in CT26 syngeneic mouse models. Mice were treated with vehicle (Veh), 60 mg / kg BID atirmociclib (At), 10 mg / kg Q3Dx3, Q10D PD1 antibody PF- 06937004 (aPD1), or combinations thereof. Each data point is the mean value from 12 mice (±SEM). Percent TGI for treatment arms is shown. Asterisk indicates independently derived proliferation IC50 (pM) for atirmociclib in CT26 cell lines in vitro.

[0049] DETAILED DESCRIPTION OF THE INVENTION

[0050] The present invention may be understood more readily by reference to the following detailed description of the embodiments and preferred embodiments of the invention, it is to be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to be limiting. It is further to be understood that unless specifically defined herein, the terminology used herein is to be given its traditional meaning as known in the relevant art.

[0051] Methods

[0052] El A method of treating a cancer in a subject, as defined above.

[0053] E2 The method of embodiment 1, wherein the CDK4 inhibitor of Formula (I) is atirmociclib.

[0054] E3 The method of any one of the preceding embodiments, wherein the CDK2 inhibitor is PF-07104091.

[0055] E4 The method of any one of the preceding embodiments, wherein the method further comprises administering to a subject an endocrine therapy described herein.

[0056] E5 The method of any one of the preceding embodiments, wherein the PD-1 axis binding antagonist comprises a PD-1 binding antagonist, ora PD-L1 binding antagonist.

[0057] E6 The method of any one of the preceding embodiments, wherein the PD-1 axis binding antagonist comprises a PD-1 binding antagonist. E7 The method of any one of the preceding embodiments, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to its ligand binding partners.

[0058] E8 The method of any one of the preceding embodiments, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-Li.

[0059] E9 The method of any one of the preceding embodiments, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L2.

[0060] E10 The method of any one of the preceding embodiments, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to both PD-L1 and PD-L2,

[0061] E11 The method of any one of the preceding embodiments, wherein the PD-1 binding antagonist is an anti-PD-1 antibody,

[0062] E12 The method of any one of the preceding embodiments, wherein the PD-1 antagonist is nivolumab (MDX 1106), pembrolizumab (MK-3475), pidilizumab (CT-011), cemiplimab (REGN2810), tislelizumab (BGB-A317), spartalizumab (PDR001), sasanlimab (RN888), mAb15, MEDI-0680 (AMP-514), BGB-108, AGEN-2034, ora combination thereof.

[0063] E13 The method of any one of the preceding embodiments, wherein the anti-PD-1 antibody is sasanlimab.

[0064] E14 The method of any one of the embodiments E1-E5, wherein the PD-1 axis binding antagonist comprises a PD-L1 binding antagonist.

[0065] E15 The method of any one of the preceding embodiments, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1.

[0066] E16 The method of any one of the preceding embodiments, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1.

[0067] E17 The method of any one of the preceding embodiments, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1.

[0068] E18 The method of any one of the preceding embodiments, wherein the PD-L1 binding antagonist is an anti-PD-L1 antibody.

[0069] El 9 The method of any one of the preceding embodiments, wherein the anti-PD-L1 antibody is BMS-936559 (MDX-1105), AMP-714, atezolizumab (Tecentriq), durvalumab (imfinzi), avelumab (Bavencio), envafolimab (KN035), toripalimab (Loqtorzi), retifanlimab (Zynyz), dostarlimab (Jemperli), IBI318 (LY3434172), IMM2510, or an antibody comprising a VH region produced by the expression vector with ATCC Accession No. PTA-121183 and having the VL region produced by the expression vector with ATCC Accession No. PTA-121182, or a combination thereof.

[0070] E20 The method of any one of the preceding embodiments, wherein the subject is a human.

[0071] E21 The method of any one of the preceding embodiments, wherein the cancer is a solid tumor. E22 The method of any one of the preceding embodiments, wherein the cancer is selected from the group consisting of brain cancer, head / neck cancer (including squamous cell carcinoma of the head and neck (SCCHN)), prostate cancer, ovarian cancer, biadder cancer (including urothelial carcinoma, also known as transitional cell carcinoma (TCC)), lung cancer (including squamous cell carcinoma, small cell lung cancer (SCLC), and non-small cell lung cancer (NSCLC)), breast cancer, bone cancer, colorectal cancer, kidney cancer, liver cancer (including hepatocellular carcinoma (HCC)), stomach cancer, pancreatic cancer, esophageal cancer, cervical cancer, sarcoma, skin cancer (including melanoma and Merkel cell carcinoma (MCC)), mesothelioma, malignant rhabdoid tumors, neuroblastoma, diffuse intrinsic pontine glioma (DIPG), and carcinoma.

[0072] E23 The method of any one of the preceding embodiments, wherein the cancer is breast cancer.

[0073] E24 The method of any one of the preceding embodiments, wherein the breast cancer is selected from any one or more of: hormone receptor positive (HR+), hormone receptor negative (HR-), human epidermal growth factor receptor 2 negative (HER2-), human epidermal growth factor receptor 2 positive (HER2+), HR+ / HER2-, ER- / HR+, ER+ / HER2- and triple negative breast cancer (TNBC).

[0074] E25 The method of any one of the embodiments E1-E22, wherein the cancer is NSCLC, prostate, colorectal cancer, liposarcoma, or tumors characterized by amplification or overexpression of CDK4 and / or CCND1.

[0075] E26 The method of any one of the embodiments E1-E22 and E25, wherein the cancer is NSCLC.

[0076] E27 The method of any one of the embodiments E1-E22 and E25, wherein the cancer is colorectal cancer.

[0077] E28 The method of any one of the preceding embodiments, wherein the method further comprise administering chemotherapy, radiotherapy, immunotherapy, or phototherapy, or any combinations thereof to the subject.

[0078] Combinations

[0079] E30 A combination(s) for use in the treatment of cancer in a subject, comprising:

[0080] (i) a CDK4 inhibitor having a Formula (I):

[0081] H.

[0082] N

[0083]

[0084] wherein:

[0085] R1is H, F or Cl;

[0086] R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R5;

[0087] R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by Rs;

[0088] R4is H or F; and

[0089] each R5and R6is independently OH, F orCi-C2alkoxy;

[0090] (ii) a PD-1 axis binding antagonist; and

[0091] (iii) optionally tegtociclib.

[0092] E31 The combination of embodiment E30, wherein the CDK4 inhibitor of Formula (I) is atirmociclib.

[0093] E32 The combination of any one of embodiments E30-E31, wherein the PD-1 axis binding antagonist is a PD-1 binding antagonist.

[0094] E33 The combination of any one of embodiments E30-E32, wherein the PD-1 binding antagonist is an anti-PD-1 antibody.

[0095] E34 The combination of any one of embodiments E30-E33, wherein the anti-PD-1 antibody is nivolumab (MDX 1106), pembrolizumab (MK-3475), pidilizumab (CT-011), cemiplimab (REGN2810), tislelizumab (BGB-A317), spartalizumab (PDR001), RN888 (sasanlimab), mAb15, MEDI-0680 (AMP-514), BGB-108, AGEN-2034, ora combination thereof.

[0096] E35 The combination of any one of the embodiments E30-E31, wherein the PD-1 axis binding antagonist is a PD-L.1 binding antagonist.

[0097] E36 The combination of any one of the embodiments E30-E31 and E35, wherein the PD-L1 binding antagonist is an anti-PD-L1 antibody.

[0098] E37 The combination of any one of the embodiments E30-E31 and E35-36, wherein the anti-PD-L1 antibody is BMS-936559 (MDX-1105), AMP-714, atezolizumab (Tecentriq), durvalumab (Imfinzi), avelumab (Bavencio), envafolimab (KN035), toripalimab (Loqtorzi), retifanlimab (Zynyz), dostarlimab (Jemperli), IBI318 (LY3434172), IMM2510, or an antibody comprising a VH region produced by the expression vector with ATCC Accession No. PTA- 121183 and having the VL region produced by the expression vector with ATCC Accession No. PTA-121182, or a combination thereof.

[0099] E38 The combination of any one of the embodiments E30-E37, wherein the combination is synergistic.

[0100] E39 The combination of any one of the embodiments E30-E38, wherein the subject is a human.

[0101] E40 The combination of any one of the embodiments E30-E39, wherein the cancer is a solid tumor.

[0102] E41 The combination of any one of the embodiments E30-E40, wherein the cancer is selected from the group consisting of brain cancer, head / neck cancer (including squamous cell carcinoma of the head and neck (SCCHN)), prostate cancer, ovarian cancer, bladder cancer (including urothelial carcinoma, also known as transitional cell carcinoma (TCC)), lung cancer (including squamous ceil carcinoma, small cell lung cancer (SCL. C), and non-small cell lung cancer (NSCLC)), breast cancer, bone cancer, colorectal cancer, kidney cancer, liver cancer (including hepatocellular carcinoma (HCC)), stomach cancer, pancreatic cancer, esophageal cancer, cervical cancer, sarcoma, skin cancer (including melanoma and Merkel cell carcinoma (MCC)), mesothelioma, malignant rhabdoid tumors, neuroblastoma, diffuse intrinsic pontine glioma (DIPG), and carcinoma.

[0103] E42 The combination of any one of the embodiments E30-E41, wherein the cancer is breast cancer.

[0104] E43 The combination of any one of the embodiments E30-E42, wherein the breast cancer is selected from any one or more of: hormone receptor positive (HR+), hormone receptor negative (HR-), human epidermal growth factor receptor 2 negative (HER2-), human epidermal growth factor receptor 2 positive (HER2+), HR+ / HER2-, ER- / HR+, ER+ / HER2- and triple negative breast cancer (TNBC).

[0105] E44 The combination of any one of the embodiments E30-E41, wherein the cancer is NSCLC, prostate, colorectal cancer, liposarcoma, or tumors characterized by amplification or overexpression of CDK4 and / or CCND1.

[0106] E45 The combination of any one of the embodiments E30-E41 and E44, wherein the cancer is NSCLC.

[0107] E46 The combination of any one of the embodiments E30-E41 and E44, wherein the cancer is colorectal cancer.

[0108] Each of the embodiments described below can be combined with any other embodiment described herein which is not inconsistent with the embodiment(s) with which it is combined. Furthermore, each of the embodiments described herein envisions within its scope pharmaceutically acceptable salts of the small molecule compounds described herein. Accordingly, the phrase “or a pharmaceutically acceptable salt thereof” is implicit in the description of all small molecule compounds described herein.

[0109] The present invention may be understood more readily by reference to the following detailed description of the preferred embodiments of the invention and the Examples included herein. It is to be understood that the terminology used herein is for the purpose of describing specific embodiments only and Is not intended to be limiting. It is further to be understood that unless specifically defined herein, the terminology used herein is to be given its traditional meaning as known in the relevant art.

[0110] As used herein, the singular form "a," "an," and "the" include plural references unless indicated otherwise. For example, "a" substituent includes one or more substituents. Where the plural form is used for compounds, salts, and the like, this is taken to mean also a single compound, salt, or the like. The invention described herein suitably may be 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 with either of the other two terms.

[0111] The term “about” when used to modify a numerically defined parameter (e.g., the dose of a CDK inhibitor, the dose of a PD-1 axis binding antagonist, means that the parameter may vary by as much as 10% above or below the stated numerical value for that parameter. For example, a dose of about 5 mg / kg should be understood to mean that the dose may vary between 4.5 mg / kg and 5.5 mg / kg.

[0112] CDK4 Inhibitors

[0113] The invention relates to methods, combinations, and uses comprising a CDK4 inhibitor, wherein the CDK4 inhibitor is a compound of Formula (I), which includes hydrates (e.g., monohydrate), salts, and polymorphs thereof:

[0114]

[0115] wherein:

[0116] R1is H, F or Cl;

[0117] R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by Rs;

[0118] R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by Rs;

[0119] R4is H or F; and

[0120] each R5and R6is independently OH, F or Ci-C2alkoxy.

[0121] In each instance recited herein, reference to “a compound of Formula (I)” may be replaced by “a CDK4 inhibitor of Formula (I).”

[0122] In one embodiment, CDK4 selective inhibitors of the present invention include atirmociclib which refers to 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-y!)- 1H-benzimidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol (also refers to as PF- 07220060), which has the following chemical structure, including hydrates (e.g., monohydrate), salts and polymorphs thereof:

[0123]

[0124] In one embodiment, the compound of Formula (I) is 1,5-anhydro-3-({5-chloro-4- 4-fluoro- 2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benz-imidazol-6-yl]pyrimidin-2-yl}amino)-2,3- dideoxy-D-threo-pentitol or a pharmaceutically acceptable salt thereof.

[0125] In one embodiment, 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1- (propan-2-yl)-1 H-benz-imidazol-6-y pyrimidin-2"yl}amino)-2,3-dideoxy" D-threo-pentitol is 1,5-anhydro-3-({5-chloro-4-[4-fluoro-2-(2-hydroxypropan-2-yl)-1-(propan-2-yl)-1H-benz-imidazol-6-yl]pyrimidin-2-yl}amino)-2,3-dideoxy-D-threo-pentitol monohydrate.

[0126] The preparation of compounds of Formula (I), including atirmociclib, are described in WO 2019 / 207463 and U. S. Patent No. 10,766,884, the contents of which are incorporated herein by reference in their entirety.

[0127] Atirmociclib is disclosed in international Publication No. WO 2019 / 207463, U. S. Patent No. 10,766,884, and International Publication No. WO 2022 / 058871, the contents of which are incorporated herein by reference in their entirety. Unless indicated otherwise, all references herein to atirmociclib include references to salts, solvates, hydrates, and complexes thereof, and to solvates, hydrates and complexes of salts thereof, including polymorphs, stereoisomers, and isotopicaliy labelled versions thereof.

[0128] Atirmociclib can be prepared as described in Example A94 of U. S. Patent No. 10,766,884 and as described in WO 2022 / 058871, the contents of which are incorporated herein by reference in their entirety.

[0129] Tegtociclib

[0130] Tegtociclib (PF-07104091) or (1R,3S)-3-[3-({ 3-(methoxymethyl)-1-methyl-1H-pyrazol-5- yl]carbonyl}amino)-1H-pyrazol-5-yl]cyclopentyl propan-2-ylcarbamate (also refers to as PF- 07104091) is a potent and selective inhibitor of cyclin dependent kinase 2 (CDK2), having the structure:

[0131]

[0132] Tegtociclib is currently in clinical development for the treatment of certain cancers. Preparation of tegtociclib is disclosed in International Patent Publication No. WO 2020 / 157652 and in United States Patent No. 11,014,911, the contents of each which are incorporated herein by reference in their entirety.

[0133] Palbociclib

[0134] Palbociclib, or 6-acetyl-8-cyclopentyl-5-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-8H-pyrido[2,3-d]pyrimidin-7-one (also referred to as “palbo”) is a potent and selective inhibitor of CDK4 and CDK6 (a CDK4 / 6 inhibitor), having the structure:

[0135]

[0136] Palbociclib and pharmaceutically acceptable salts thereof, are disclosed in International Publication No. WO 2003 / 062236 and U. S. Patent Nos. 6,936,612, 7,208,489 and 7,456,168; International Publication No. WO 2005 / 005426 and U. S. Patent Nos. 7,345,171 and 7,863,278; International Publication No. WO 2008 / 032157 and U. S. Patent No. 7,781,583; and International Publication No. WO 2014 / 128588. The contents of each of the foregoing references are incorporated herein by reference in their entirety.

[0137] PD-1 Axis Binding Antagonists

[0138] As used herein, the term “PD-1 axis binding antagonist” or “PD-1 axis antagonist” refers to a molecule that inhibits the interaction of a PD-1 axis binding partner (e.g., PD-1, PD-L1, PD- L2) with either one or more of its binding partners, for example so as to overcome or partially overcome T-cell dysfunction resulting from signaling on the PD-1 signaling axis— with a result being to restore, partially restore or enhance T-cell function (e.g., proliferation, cytokine production, target ceil killing, survival). As used herein, a PD-1 axis binding antagonist includes one or more of (i) a PD-1 binding antagonist, (ii) a PD-L1 binding antagonist, and / or (iii) a PD-L2 antagonist.

[0139] The term “PD-1 binding antagonist,” as used herein, refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-1 with one or more of its binding partners, such as PD-L1, PD-L2. in some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its binding partners. In a specific aspect, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2 For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In some embodiments, a PD-1 binding antagonist reduces the negative co-stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-1 so as to render a dysfunctional T-cell less non-dysfunctional.

[0140] Exemplary PD-1 binding antagonists include those described in U. S. Patent Application Publication 20130280265, U. S. Patent Application Publication 20130237580, U. S. Patent Application Publication 20130230514, U. S. Patent Application Publication 20130109843, U. S. Patent Application Publication 20130108651, U. S. Patent Application Publication 20130017199, U. S. Patent Application Publication 20120251537, U. S. Patent Application Publication 20110271358, European Patent EP2170959B1, in PCT Publication No. WO 2011 / 066342, PCT Publication No. WO 2015 / 035606, PCT Publication No. WO 2015 / 085847, PCT Publication No. WO 2015 / 112800, PCT Publication No. WO 2015 / 112900, PCT Publication No. WO 2016 / 092419, PCT Publication No. WO 2017 / 017623, PCT Publication No. WO 2017 / 024465, PCT Publication No. WO 2017 / 054646, PCT Publication No. WO 2017 / 071625, PCT Publication No. WO 2017 / 019846, PCT Publication No. WO 2017 / 132827, PCT Publication No. WO 2017 / 214092, PCT Publication No. WO 2018 / 013017, PCT Publication No. WO 2018 / 053106, PCT Publication No. WO 2018 / 055503, PCT Publication No. WO 2018 / 053709, PCT Publication No. WO 2018 / 068336, and PCT Publication No. WO 2018 / 072743, the entire disclosures of which are incorporated herein by reference. Other exemplary PD-1 binding antagonists are described in Curran etal., PD-1 and CTLA-4 combination blockade expands infiltrating T cells and reduces regulatory T and myeloid cells within 816 melanoma tumors, PNAS, 2010, 107, 4275; Topalian etal., Safety, activity, and immune correlates of anti-PD-1 antibody in cancer, New Engl. J. Med.

[0141] 2012, 366, 2443; Brahmer ef a / ., Safety and activity of anti-PD-L1 antibody in patients with advanced cancer, New Engl. J. Med. 2012, 366, 2455; Dolan et al., PD-1 pathway inhibitors: changing the landscape of cancer immunotherapy, Cancer Control 2014, 21, 3; and Sunshine ef a / ., Pd-1 / Pd-L1 Inhibitors, Curr. Opin. in Pharmacol. 2015, 23.

[0142] The term “PD-L1 binding antagonist,” as used herein, refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-L1 with either one or more of its binding partners, such as PD-1, B7-1. in some embodiments, the PD-1 axis binding antagonist comprises a PD-L1 binding antagonist. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to its binding partners, in a specific aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1. In another specific aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In another specific aspect, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7- 1.

[0143] In some embodiments, the PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1, and / or B7- 1. In some embodiments, a PD-L1 binding antagonist reduces the negative co-stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-L1 so as render a dysfunctional T-cell less non-dysfunctional. In some embodiments, a PD-L1 binding antagonist is an anti-PD-L1 antibody (aPD-L1). In some embodiments, the PD- L1 antibody is a biosimiiar, biobetter, or bioequivalent thereof.

[0144] Some exemplary PD-L1 binding antagonists include those described in U. S. Patent Application Publication 20090055944, U. S. Patent Application Publication 20100203056, U. S. Patent Application Publication 20120039906, U. S. Patent Application Publication 20130045202, U. S. Patent Application Publication 20130309250, U. S. Patent Application Publication US20130034559, U. S. Patent Application Publication US20150282460, U. S. Patent Application Publication 20160108123, PCT Publication No. WO 2011 / 066389, PCT Publication No. WO 2016 / 000619, PCT Publication No. WO 2016 / 094273, PCT Publication No. WO 2016 / 061142, PCT Publication No. WO 2016 / 149201, PCT Publication No. WO 2016 / 149350, PCT Publication No. WO 2016 / 179576, PCT Publication No. WO 2017 / 020801, PCT Publication No. WO 2017 / 103147, PCT Publication No. WO 2017 / 112741, PCT Publication No. WO 2017 / 205213, PCT Publication No. WO 2017 / 054646, PCT Publication No. WO 2017 / 084495, PCT Publication No. WO 2017 / 161976, PCT Publication No. WO 2018 / 005682, PCT Publication No. WO 2018 / 053106, PCT Publication No. WO 2018 / 085469, PCT Publication No. WO 2018 / 111890, and PCT Publication No. WO 2018 / 106529, the entire disclosures of which are incorporated herein by reference. Other exemplary PD-L1 binding antagonists are described in Sunshine ef a / ., 2015.

[0145] The term “PD-L2 binding antagonist,” as used herein, refers to a molecule that decreases, blocks, inhibits, abrogates or interferes with signal transduction resulting from the interaction of PD-L2 with either one or more of its binding partners, such as PD-1. in some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its binding partners. In a specific aspect, the PD-L2 binding antagonist inhibits binding of PD-L2 to PD-1. In some embodiments, the PD-L2 antagonists include anti-PD-L2 antibodies, antigen binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides and other molecules that decrease, block, inhibit, abrogate or interfere with signal transduction resulting from the interaction of PD- L2 with either one or more of its binding partners, such as PD-1. in some embodiments, a PD-L2 binding antagonist reduces the negative co-stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes mediated signaling through PD-L2 so as render a dysfunctional T-cell less non-dysfunctional. In some embodiments, a PD-L2 binding antagonist is a PD-L2 immunoadhesin.

[0146] In some embodiments the PD-1 axis binding antagonist (e.g., PD-1 binding antagonist, PD-L1 binding antagonist, or PD-L2 binding antagonist) is a small molecule antagonist. In some further embodiments the PD-1 axis binding antagonist (e.g., PD-L1 binding antagonist) is a chemical compound disclosed in PCT Publication No. WO 2015 / 033299 or PCT Publication No. WO 2015 / 033301 or a pharmaceutically acceptable salt thereof.

[0147] Table 1 below provides a list of the amino acid sequences of exemplary PD-1 axis binding antagonists for use in the treatment method, medicaments and uses of the present invention. CDRs are underlined for mAb7 and mAb15. The mAB7 is also known as RN888 or PF-06801591. mAb7 and mAb15 are disclosed in PCT Publication No. WO 2016 / 092419, the disclosure of which is hereby incorporated by reference in its entirety.

[0148] Table 1. Exemplary PD-1 axis binding antagonists

[0149] mAb7(aka RN888) or QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWNWVRQAPGQG I mAb15 full-length LEWMGNIYPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSE heavy chain DTAVYYCARLSTGTFAYWGQGTLVTVSSASTKGPSVFPLAPCSRS WO 2016 / 092419 TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1)

[0150] m. Ab7 or mAbl 5 full- QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWNWVRQAPGQG | length heavy chain LEWMGNIYPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSE without the C-terminal DTAVYYCARLSTGTFAYWGQGTLVTVSSASTKGPSVFPLAPCSRS lysine TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY WO 2016 / 092419 SLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP

[0151]

[0152] CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 2)

[0153] mAb7 full-length light DIVMTQSPDSLAVSLGERATINCKSSQSLWDSGNQKNFLTWYQQK | chain PGQPPKLLIYWTSYRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV WO 2016 / 092419 YYCQNDYFYPHTFGGGTKVEIKRGTVAAPSVFIFPPSDEQLKSGTA SWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 3)

[0154] mAb7 light chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWNWVRQAPGQG I variable region LEWMGNIYPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSE WO 2016 / 092419 DTAVYYCARLSTGTFAYWGQGTLVTVSS (SEQ ID NO: 4) mAB7 and mAB15 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWINWVRQAPGQG heavy chain variable LEWMGNIWPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSE region DTAVYYCARLLTGTFAYWGQGTLVTVSS (SEQ ID NO: 5)

[0155] WO 2016 / 092419

[0156] mAb15 light chain DIVMTQSPDSLAVSLGERATINCKSSQSLWDSGNQKNFLTWYQQK | variable region PGQPPKLLIYWTSYRESGVPDRFSGSGSGTDFTLTISSLQAEDVAV WO 2016 / 092419 YYCQNDYFYPHTFGGGTKVEIK (SEQ ID NO: 6)

[0157] Nivolurnab, QVQLVESGGGWQPGRSLRLDCKASGITFSNSGMHWVRQAPGKG MDX1106, full length LEWVAVrWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAE heavy chain DTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSES WO 2006 / 121168 TAALGCLVDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTTYTCNVDHKPSNTKVDRVESYGPPCPPCPAPEFLG GPSVFLFPPKPKDTLMISRTPEVTCWVDVSQEDPEVQFNWYYDGV EVHNATKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKA GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPEKNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTQKSLSLSLGK (SEQ ID NO: 7)

[0158] Nivolurnab, EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQPGQAPRLL | MDX1106, full length IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSN light chain WPRTFGQGTKVEIRTVAAPSVFIFPPSDEQLSGTASVVCLLNNFYP WO 2006 / 121168 REAVQWKVDNALQSGNSQESVTEQDSDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVT SFNRGEC (SEQ ID NO: 8)

[0159]

[0160] Pembrolizumab, QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPG MK3475, full length QGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKS heavy chain LQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPS WO 2009 / 114335 VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVD KRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMSSRTPEV TCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPRE PQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEA LHNHYTQKSLSLSLGK (SEQ ID NO: 9)

[0161] Pembrolizumab, EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKP | MK3475, full length GQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAV light chain YYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTA WO 2009 / 114335 SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTY SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC

[0162] (SEQ ID NO: 10)

[0163] AMP-224, without LFTVTVPKELYIIEHGSNVTLECNFDTGSHVNLGAITASLQKVENDTS signal sequence PHRERATLLEEQLPLGKASFHIPQVQVRDEGQYQCIIIYGVA WO 2010 / 027827 & WDYKYLTLKVKASYRKI NTH I LKVPETDEVELTCQATGYPLAEVSW WO 2011 / 066342 PNVSVPANTSHSRTPEGLYQVTSVLRLKPPPGRNFSCVFWNTHVR ELTLASIDLQSQMEPRTHPTWEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGFY PSDIAVEWES NGQPENNYKT TPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0164] (SEQ ID NO: 11)

[0165] YW243.55. S70 heavy EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKG I chain variable region LEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAE WO 2010 / 077634 DTAVYYCARRHWPGGFDYWGQGTLVTVSA (SEQ ID NO: 12)

[0166] YW243.55. S70 light DSQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPK chain variable region LLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYL WO 2010 / 077634 YH PATFGQGTKVEIKR (SEQ ID NO: 13)

[0167] avelumab heavy EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMWVRQAPGKGLE | chain variable region WVSSSYPSGGITFYADKGRFTISRDNSKNTLYLQMNSLRAEDTAVY WO 13079174 YCARIKLGTVTTVDYWGQGTLVTVSS (SEQ ID NO: 14)

[0168]

[0169] aveiumab light chain QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKA variable region PKLMIYDVSNRPSGVSNRFSGSKSGNTAS LTISGLQAEDEADYYCS WO 2013 / 079174 SYTSSSTRVFGTGTKVTVL (SEQ ID NO: 15)

[0170] AGEN-2034W QVQLVESGGGWQPGRSLRLSCAASGFTFSSYGMHWVRQAPGK Full length heavy GLEWVAVIWYDGSNKYYADSVKGRFTiSRDNSKNTLYLQMNSLRA chain EDTAVYYCASNGDHWGQGTLVTVSSASTKGPSVFPLAPCSRSTSE CAS RN: 2088287- STAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS 86-7 SWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPA WO 2017 / 040790 PEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNW YVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDK SRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 16) AGEN2034W EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPR 1 full length light chain LLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYN CAS RN: 2088287- NWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNN 75-4 FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL. SK WO 2017 / 040790 ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 17) Spartaiizumab EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQG Full length heavy LEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSED chain TAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRST CAS RN: 1935694- SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY 88-4 SLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP WO / 2015 / 112900 CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 18)

[0171] Spartaiizumab Full EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKP length light chain GQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATY CAS RN: 1935694- YCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV 88-4 VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS WO 2015 / 112900 TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:

[0172] 19)

[0173] Cemipiimab (REGN- EVQLLESGGVLVQPGGSLRLSCAASGFTFSNFGMTWVRQAPGKG | 2810) LEWVSGISGGGRDTYFADSVKGRFTISRDNSKNTLYLQMNSLKGE

[0174] DTAVYYCVKWGNIYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRS

[0175]

[0176] CAS RN: 1801342- TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY 60-8 SLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP Full length heavy CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQ chain FNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKE WO 2015 / 112800 YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRL TVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 20)

[0177] Cemiplimab (REGN- DIQMTQSPSSLSASVGDSITITCRASLSINTFLNWYQQKPGKAPNLLI | 2810) YAASSLHGGVPSRFSGSGSGTDFTLTIRTLQPEDFATYYCQQSSNT CAS RN: 1801342- PFTFGPGTVVDFRRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY 60-8 PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD full length light chain YEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 21) WO 2015 / 112800

[0178] Durvalumab EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGK (IMFINZI®) GLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRA CAS RN: 2222916- EDTAVYYCAREGGWFGELAFDYWGQGTLVTVSSASTKGPSVFPL 00-7 APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV Full length heavy LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS chain CDKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVWD WO 2011 / 066389 & VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLH WO 2018 / 106529 QDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0179] K (SEQ ID NO: 22)

[0180] Durvalumab (MEDI- EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAP | 4736 or IMFINZI ®) RLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQY CAS RN: 2222915- GSLPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLN 99-1 NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS full length light chain KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 23) WO 2011 / 066389 &

[0181] WO 2018 / 106529

[0182] Atezoiizumab EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKG CAS number: LEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAE 1380723-44-3 DTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSS Full length heavy KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS chain GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT WO 2018 / 106529 HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE

[0183]

[0184] DPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0185] (SEQ ID NO: 24)

[0186] Atezolizumab DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPK full length light chain LLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYL WO 2018 106529 YHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 25) Envafolimab (KN035) QVQLVESGGGLVQPGGSLRLSCAASGFTFSRRCMAWFRQAPGKE I RERVAKLLTTSGSTYLADSVKGRFTISRDNSKNTVYLQMNSLRAED

[0187] Single domain TAVYYCAADSFEDPTCTLVTSSGAFQYWGQGTLVTVSS (SEQ ID antibody NO: 26)

[0188] From European

[0189] Publication No,

[0190] EP3330290A1, (aka,

[0191] hu56V2)

[0192] BMS-936559 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQGL Heavy chain variable EWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTA region VYFCARKFHFVSGSPFGMDVWGQGTTVTVSS (SEQ ID NO: 27) WO 2018 / 106529 &

[0193] WO 2007 / 005874

[0194] BMS-936559 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRL I Light chain variable LIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSN region WPTFGQGTKVEIK (SEQ ID NO: 28)

[0195] WO 2018 / 106529 &

[0196] WO 2007 / 005874

[0197] Pidilizumab QVQLVQSGSELKKPGASVKISCKASGYTFTNY GMNWVRQAPGQG Full length heavy LQWMGWINTDSGESTYAEEFKGRFVFSLDTSVNTAYLQITSLTAED chain TGMYFCVRVGYDALDYWGQGTLVTVSSASTKGPSVFPLAPSSKST WO 2009 / 101611 SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS

[0198]

[0199] KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 29)

[0200] Pidilizumab EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWFQQKPGKAPKL Full length light chain WIYRTSNLASGVPSRFSGSGSGTSYCLTINSLQPEDFATYYCQQRS WO 2009 / 101611 SFPLTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA DYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 30) Tislelizumab QVQLQESGPGLVKPSETLSLTCTVSGFSLTSYGVHWIRQPPGKGL I CAS Registry EWIGVIYADGSTNYNPSLKSRVTISKDTSKNQVSLKLSSVTAADTAV Number: 1858168- YYCARAYGNYWYIDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTS 59-8 ESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL Full length heavy SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCP chain APPVAGGPSVFLFPPKPKDTLMISRTPEVTCVWAVSQEDPEVQFN WO 2015 / 035606 WYVDGVEVHNAKTKPREEQFNSTYRVVSVLTWHQDWLNGKEYK CKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 31)

[0201] Tislelizumab DIVMTQSPDSLAVSLGERATINCKSSESVSNDVAWYQQKPGQPPK CAS Registry LLINYAFHRFTGVPDRFSGSGYGTDFTLTISSLQAEDVAVYYCHQA Number: 1858168- YSSPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN 59-8 FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK Full length light chain ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 32) WO 2015 / 035606

[0202]

[0203] As used herein, an anti-human PD-L1 mAb refers to a monoclonal antibody that specifically binds to mature human PD-L1. A mature human PD-L1 molecule consists of amino acids 19-290 of the following sequence MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWeAEMEDKNII QFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITV KVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFN VTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFR LRKGRMMDVKKCGIQDTNSKKQSDTHLEET (SEQ ID NO: 33).

[0204] Table 2 below provides exemplary anti-PD-L1 antibody sequences for use in the treatment method, medicaments and uses of the present invention. Table 2. Exemplary anti-human PD-L1 monoclonal antibody sequences

[0205] Heavy chain SYIMM (SEQ ID NO: 34)

[0206] CDR1 (CDRH1)

[0207] Heavy chain SIYPSGGITFY (SEQ ID NO: 35)

[0208] CDR2 (CDRH2)

[0209] Heavy chain IKLGTVTTVDY (SEQ ID NO: 36) i CDR3 (CDRH3)

[0210] Light chain CDR1 TGTSSDVGGYNYVS (SEQ ID NO: 37)

[0211] (CDRL1)

[0212] Light chain CDR2 DVSNRPS (SEQ ID NO: 38)

[0213] (CDRL2)

[0214] Light chain CDR3 SSYTSSSTRV (SEQ ID NO: 39) i (CDRL3)

[0215] Heavy chain EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEW I variable region VSSIYPSGGITFYADKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARI (VR) KLGTVTTVDYWGQGTLVTVSS (SEQ ID NO: 14)

[0216] QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPK

[0217] Light chain VR LMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTS SSTRVFGTGTKVTVL (SEQ ID NO: 15) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEW | VSSIYPSGGITFYADTVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYC ARIKLGTVTTVDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVP SSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG

[0218] Heavy chain

[0219] PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGK (SEQ ID NO: 40) QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPK LMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTS

[0220] Light chain SSTRVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFY PGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 41)

[0221]

[0222] In some embodiments, the PD-1 axis binding antagonist is RN888 and will be administered subcutaneously at a dose of about 1, 2, 3, 4, 5, 6, 7 or 8 mg / kg at intervals of about 14 days (± 2 days) or about 21 days (± 2 days) or about 30 days (+ 2 days) throughout the course of treatment. In some embodiment, RN888 is administer as a flat dose of about 80, 150, 160, 200, 240, 250, 300, 320, 350, 400, preferably 300mg at intervals of about 14 days (± 2 days) or about 21 days (± 2 days) or about 30 days (± 2 days). In some embodiments, RN888 is administered subcutaneously in an amount of 300 mg Q4W.

[0223] In one embodiment, " PD-1 antagonist" means any chemical compound or biological molecule that blocks binding of PD-L1 expressed on a cancer cell to PD-1 expressed on an immune cell (T cell, B cell or NKT cell) and preferably also blocks binding of PD-L2 expressed on a cancer cell to the immune-cell expressed PD-1. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279 and SLEB2 for PD-1; PDCD1L1, PDL1, B7H1, B7-4, CD274 and B7-H for PD-L1; and PDCD1L2, PDL2, B7-DC, Btdc and CD273 for PD-L2. In any of the treatment method, medicaments and uses of the present invention in which a human individual is being treated, the PD-1 antagonist may block binding of human PD-L1 to human PD- 1, and block binding of both human PD-L1 and PD-L2 to human PD-1. Exemplary human PD-1 amino acid sequences can be found in NCBI Locus No.: NP_005009. Exemplary human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

[0224] PD-1 antagonists useful in any of the treatment methods, medicaments and uses of the present invention include a monoclonal antibody (mAb), or antigen binding fragment thereof, which specifically binds to PD-1 or PD-L1, and preferably specifically binds to human PD-1 or human PD-L1, The mAb may be a human antibody, a humanized antibody or a chimeric antibody, and may include a human constant region. In some embodiments the human constant region is selected from the group consisting of IgG 1, lgG2, lgG3 and lgG4 constant regions, and in some embodiments, the human constant region is an lgG1 or lgG4 constant region. In some embodiments, the antigen binding fragment is selected from the group consisting of Fab, Fab'-SH, F(ab')2, scFv and Fv fragments.

[0225] Examples of mAbs that bind to human PD-1, and useful in the treatment method, medicaments and uses of the present invention, are described in U. S. Patent Nos. 7,488,802, 7,521,051, 8,008,449, 8,354,509, 8,168,757, PCT Patent Publication Nos. WO 2004 / 004771, WO 2004 / 072286, WO 2004 / 056875, and US Patent Publication No. 2011 / 0271358.

[0226] Table 3 below provides exemplary anti-PD-1 antibody sequences for use in the treatment method, medicaments and uses of the present invention.

[0227] Table 3 Exemplary anti-human PD-1 monoclonal antibody (RN888) sequences

[0228] Heavy chain GYTFTSYWIN (SEQ ID NO: 42)

[0229] CDR1 (CDRH1)

[0230] Heavy chain i NIYPGSSLTNYNEKFKN (SEQ ID NO: 43)

[0231] CDR2 (CDRH2)

[0232] Heavy chain LSTGTFAY (SEQ ID NO: 44)

[0233] CDR3 (CDRH3)

[0234]

[0235] Light chain CDR1 i KSSQSLWDSGNQKNFLT (SEQ ID NO: 45)

[0236] (CDRL1) |

[0237] Light chain CDR2 i WtSYRES (SEQ ID NO: 46)

[0238] (CDRL2) |

[0239] Light chain CDR3 QNDYFYPHT (SEQ ID NO: 47)

[0240] (CDRL3) i

[0241] Heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWIN

[0242] variable region | WMGNIYPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSEDTAV (VR) YYCARLSTGTFAYWGQGTLVTVSS (SEQ ID NO: 4)

[0243] Light chain VR | QPPKLLIYWTSYRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQ i NDYFYPHTFGGGTKVEIK (SEQ ID NO: 6)

[0244] | QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWINWVRQAPGQGLE | WMGNIYPGSSLTNYNEKFKNRVTMTRDTSTSTVYMELSSLRSEDTAV | YYCARLSTGTFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTA | ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT i VPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGG Heavy chain |

[0245] i PSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEV | HNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSS | IEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE | WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 1)

[0246] | DIVMTQSPDSLAVSLGERATINCKSSQSLWDSGNQKNFLTWYQQKPG | QPPKLLIYWTSYRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQ Light chain | NDYFYPHTFGGGTKVEIKRGTVAAPSVFIFPPSDEQLKSGTASVVCLL | NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK i ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 3)

[0247]

[0248] Endocrine Therapy

[0249] As used herein, “endocrine therapy” or “hormone therapy” means an aromatase inhibitor, a selective estrogen receptor degrader (SERD), or a selective estrogen receptor modulator (SERM). in certain embodiments, endocrine therapy includes fulvestrant, tamoxifen, toremifene, anastrozole, exemestane, or ietrozole.

[0250] The term “antiestrogen” as used herein refers to a class of drugs that prevent estrogens like estradiol from mediating the biological effects in the body. Antiestrogens act by blocking the estrogen receptor (ER) and / or inhibiting or suppressing estrogen production. In an embodiment, an antiestrogen is an aromatase inhibitor, a selective estrogen receptor degrader (SERD) or a selective estrogen receptor modulator (SERM). Examples of an aromatase inhibitor include, but are not limited to, anastrozole. Examples of a SERD include, but are not limited to, fuivestrant. Additional SERDs include elacestrant (RAD-1901, Radius Health), SAR439859 (Sanofi), RG6171 (Roche), AZD9833 (AstraZeneca), AZD9496 (AstraZeneca), rintodestrant (G1 Therapeutics), ZN-C5 (Zentaiis), LSZ102 (Novartis), D-0502 (Inventisbio), LY3484356 (Lilly), and SHR9549 (Jiansu Hengrui Medicine). Examples of a SERM include, but are not limited to, tamoxifen, clomifene and raloxifene. Additional SERMS include toremifene, lasofoxifene, bazedoxifene and afimoxifene.

[0251] In an embodiment, the aromatase inhibitor includes letrozole, exemestane, and anastrozole. In an embodiment, the SERM includes tamoxifen, clomifene and raloxifene.

[0252] In an embodiment, an antiestrogen of the present invention includes fuivestrant and letrozole. In an embodiment, an antiestrogen of the present invention includes fuivestrant. In an embodiment, an antiestrogen of the present invention includes letrozole.

[0253] As used herein, terms, including, but not limited to, “drug,” “agent,” “component,” “composition," “compound," “substance," “targeted agent," “targeted therapeutic agent," “therapeutic agent,” and “medicament” may be used interchangeably to refer to the small molecule compounds of the present invention, e.g., a CDK inhibitor. As used herein, terms, including, but not limited to, “drug,” “agent," “component," “composition," “compound," “substance," “targeted agent," “targeted therapeutic agent," “therapeutic agent," therapeutic antibody,” and “medicament” may be used interchangeably to refer to the antibodies of the present invention, e.g., an anti-PD-L1 antibody, an anti-PD-1 antibody, or combinations thereof.

[0254] The term “therapeutic antibody” refers to an antibody that is used in the treatment of a disease or a disorder. A therapeutic antibody may have various mechanisms of action. A therapeutic antibody may bind and neutralize the normal function of a target associated with an antigen. For example, a monoclonal antibody that blocks the activity of the protein needed for the survival of a cancer cell causes the cell's death. Another therapeutic antibody may bind and activate the normal function of a target associated with an antigen. For example, a monoclonal antibody can bind to a protein on a cell and trigger an apoptosis signal. Yet another monoclonal antibody may bind to a target antigen expressed only on diseased tissue; conjugation of a toxic payload (effective agent), such as a chemotherapeutic or radioactive agent, to the monoclonal antibody can create an agent for specific delivery of the toxic payload to the diseased tissue, reducing harm to healthy tissue. A “biologically functional fragment” of a therapeutic antibody will exhibit at least one if not some or all of the biological functions attributed to the intact antibody, the function comprising at least specific binding to the target antigen.

[0255] The therapeutic antibody may bind to any protein, including, without limitation, a PD-L1, and / or a PD-1. Accordingly, therapeutic antibodies include, without limitation, anti-PD-L1 antibodies, anti-PD-1 antibodies, or combinations thereof. " Biotherapeutic agent" means a biological molecule, such as an antibody orfusion protein, that blocks ligand / receptor signaling in any biological pathway that supports tumor maintenance and / or growth or suppresses the anti-tumor immune response.

[0256] A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethyloiomelamine; acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicines; betulinic acid; a camptothecin (including the synthetic analogue topotecan (HYCAMTIN®), CPT- 11 (irinotecan, CAMPTOSAR®), acetylcamptothecin, scopolectin, and 9- aminocamptothecin); bryostatin; pemetrexed; callystatin; CC- 1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); podophyllotoxin; podophyllinic acid; teniposide; cryptophycins (particularly cryptophycin 1 and cryptophycin 8); doiastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB 1 -TM1 ); eleutherobin; pancratistatin; TLK-286; CDP323, an oral alpha-4 integrin inhibitor; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma and calicheamicin omega! (e.g., Nicolaou et a / ., Angew. Chem Inti. Ed, Engl., 33: 183- 186 (1994)); dynemicin, including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including ADRIAMYCIN®, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin HC1 liposome injection (DOXIL®) and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine (XELODA®), an epothilone, and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, and imatinib (a 2-phenylaminopyrimidine derivative), as well as other c- it inhibitors; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamlne; demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (" Ara-C"); thiotepa; taxoids, e.g., paclitaxel (TAXOL®), albumin- engineered nanoparticle formulation of paclitaxel (ABRAXANE™), and doxetaxel (TAXOTERE®); chloranbucil; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine (VELBAN®); platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVIN®); oxaliplatin; leucovovin; vinorelbine (NAVELBINE®); novantrone; edatrexate; daunomycin; aminopterin; ibandronate; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; pharmaceutically acceptable salts, acids or derivatives of any of the above; as well as combinations of two or more of the above such as CHOP, an abbreviation fora combined therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and FOLFOX, an abbreviation for a treatment regimen with oxaliplatin (ELOXATIN™) combined with 5-FU and leucovovin.

[0257] Additional examples of chemotherapeutic agents include anti-hormonal agents that act to regulate, reduce, block, or inhibit the effects of hormones that can promote the growth of cancer, and are often in the form of systemic, or whole-body treatment. They may be hormones themselves. Examples include anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX® tamoxifen), raloxifene (EVISTA®), droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 1 1 7018, onapristone, and toremifene (FARESTON®); anti-progesterones; estrogen receptor down-regulators (ERDs); estrogen receptor antagonists such as fulvestrant (FASLODEX®); agents that function to suppress or shut down the ovaries, for example, luteinizing hormone-releasing hormone (LHRFI) agonists such as leuprolide acetate (LUPRON® and ELIGARD®), goserelin acetate, buserelin acetate and tripterelin; anti-androgens such as fiutamide, nilutamide and bicalutamide; and aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, megestrol acetate (MEGASE®), exemestane (AROMASIN®), formestanie, fadrozole, vorozole (RJVISOR®), letrozole (FEMARA®), and anastrozole (ARIMIDEX®). In addition, such definition of chemotherapeutic agents includes bisphosphonates such as clodronate (for example, BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58Q95, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); as well as troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); anti-sense oligonucleotides, particularly those that inhibit expression of genes in signaling pathways implicated in abherent cell proliferation, such as, for example, PKC-alpha, Raf, H-Ras, and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE® vaccine and gene therapy vaccines, for example, ALLOVECTIN® vaccine, LEUVECTIN® vaccine, and VAXID® vaccine; topoisomerase 1 inhibitor (e.g., LURTOTECAN®); an anti-estrogen such as fulvestrant; a Kit inhibitor such as imatinib or EXEL-0862 (a tyrosine kinase inhibitor); EGFR inhibitor such as erlotinib or cetuximab; an anti-VEGF inhibitor such as bevacizumab; arinotecan; rmRH (e.g., ABARELIX®); lapatinib and lapatinib ditosylate (an ErbB-2 and EGFR dual tyrosine kinase small molecule inhibitor also known as GW572016); 17AAG (geldanamycin derivative that is a heat shock protein (Hsp) 90 poison), and pharmaceutically acceptable salts, acids or derivatives of any of the above.

[0258] As used herein, the term "cytokine" refers generically to proteins released by one cell population that act on another cell as intercellular mediators or have an autocrine effect on the cells producing the proteins. Examples of such cytokines include lymphokines, monokines; interleukins (" ILs") such as IL- 1, IL- la, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL10, IL-1 1, IL-12, IL-13, IL-15, IL-17A-F, IL-18 to IL-29 (such as IL-23), IL-31, including PROLEUKIN® rlL-2; a tumor-necrosis factor such as TNF-a or TNF-p, TGF- I -3; and other polypeptide factors including leukemia inhibitory factor (“LIF"), ciliary neurotrophic factor (" CNTF"), CNTF-iike cytokine (" CLC"), cardiotrophin (" CT"), and kit ligand (" L").

[0259] As used herein, the term "chemokine" refers to soluble factors (e.g., cytokines) that have the ability to selectively induce chemotaxis and activation of leukocytes. They also trigger processes of angiogenesis, inflammation, wound healing, and tumorigenesis. Example chemokines include IL-8, a human homolog of murine keratinocyte chemoattractant (KC).

[0260] The terms “abnormal cell growth” and “hyperproliferative disorder” are used interchangeably in this application. “Abnormal cell growth,” as used herein, unless otherwise indicated, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition). Abnormal cell growth may be benign (not cancerous), or malignant (cancerous).

[0261] A “disorder” is any condition that would benefit from treatment with the compounds of the present invention. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the subject to the disorder in question.

[0262] The term “antibody,” as used herein, refers to an immunoglobulin molecule capable of specific binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site, located in the variable region of the immunoglobulin molecule. As used herein, the term encompasses a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a bispecific antibody, a dual-specific antibody, bifunctional antibody, a trispecific antibody, a multispecific antibody, a bispecific heterodimeric diabody, a bispecific heterodimeric IgG, a labeled antibody, a humanized antibody, a human antibody, and fragments thereof (such as Fab, Fab’, F(ab’)2, Fv), single chain (ScFv) and domain antibodies (including, for example, shark and camelid antibodies), fusion proteins comprising an antibody, any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site, and antibody like binding peptidomimetics (ABIPs). An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof), and the antibody need not be of any particular class. Depending on the antibody amino acid sequence of the constant region of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG-1, IgG-2, IgG-3, IgG-4, IgA1 and IgA2. The heavy¬ chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three- dimensional configurations of different classes of immunoglobulins are well known.

[0263] As used herein, a “bispecific antibody," “dual-specific antibody," “bifunctional antibody," “heteromultimer," "heteromultimeric complex," “bispecific heterodimeric diabody” or a “heteromultimeric polypeptide” is a molecule comprising at least a first polypeptide and a second polypeptide, wherein the second polypeptide differs in amino acid sequence from the first polypeptide by at least one amino acid residue. In some instances, the bispecific is an artificial hybrid antibody having two different heavy chain region and light chain region. Preferably, the bispecific antibody has binding specificity for at least two different ligands, antigens or binding sites. Accordingly, the bispecific antibodies can bind simultaneously two different antigens. The two antigen binding sites of a bispecific antibody bind to two different epitopes, which may reside on the same or different protein targets, e.g., tumor target.

[0264] The bispecific antibody, dual-specific antibody, bifunctional antibody, heteromultimer, heteromultimeric complex, bispecific heterodimeric diabody or the heteromultimeric polypeptide can be prepared by constructing sFv fragments with short linkers (e.g., about 3-10 residues) between the VH and VL regions such that inter-chain but not intra-chain pairing of the V regions is achieved, resulting in a bivalent fragment, i.e., fragment having two antigen-binding sites. Bispecific antibodies can be derived from full length antibodies or antibody fragments (e.g., F(ab')2bispecific antibodies). Diabodies are described more fully in, for example, EP404,097; WO 1993 / 011161; and Hollinger et al., A small bispecific antibody construct expressed as a functional single-chain molecule with high tumor cell cytotoxicity, Proc. Natl. Acad. Sci. 1993, 90:6444-6448. Bispecific antibodies are heterodimers of two "crossover" sFv fragments in which the VH and VL regions of the two antibodies are present on different polypeptide chains.

[0265] By way of non-limiting example, a bispecific antibody may comprise one antigen-binding site that recognizes an epitope on one protein (e.g., 0X40, 4-1 BB, PD-1 or PD-L1) and further comprise a second, different antigen-binding site that recognizes a different epitope on a second protein (e.g., 0X40, 4-1 BB, PD-1 or PD-L1). Generally, but not necessarily, reference to binding means specific binding.

[0266] The term "immunoglobulin" (Ig) is used interchangeably with "antibody" herein. The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. An IgM antibody consists of 5 of the basic heterotetramer units along with an additional polypeptide called a J chain, and contains 10 antigen binding sites, while IgA antibodies comprise from 2-5 of the basic 4-chain units which can polymerize to form polyvalent assemblages in combination with the J chain. In the case of IgGs, the 4-chain unit is generally about 150,000 Daltons. Each L chain is linked to an H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has at the N-terminus, a variable domain (VH) followed by three constant domains (CH) for each of the a and y chains and four CH domains for p and s isotypes. Each L chain has at the N-terminus, a variable domain (VL) followed by a constant domain at its other end. The VL is aligned with the VH and the CL is aligned with the first constant domain of the heavy chain (CHI). Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains. The pairing of a VH and VL together forms a single antigen¬ binding site. For the structure and properties of the different classes of antibodies, e.g., Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Basic and Clinical Immunology, 8th Edition, 1994, page 71 and Chapter S. The L chain from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains (CH), immunoglobulins can be assigned to different classes or isotypes.

[0267] The terms "full-length antibody,” "intact antibody" or "whole antibody" are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antibody fragment. Specifically, whole antibodies include those with heavy and light chains including an Fc region. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, the intact antibody may have one or more effector functions.

[0268] An "antibody fragment" comprises a portion of an intact antibody, preferably the antigen binding and / or the variable region of the intact antibody. Examples of antibody fragments suitable for use in this invention include, without limitation: (i) the Fab fragment, consisting of VL, VH, CL, and CH1 domains; (ii) the “Fd” fragment consisting of the VH and CH1 domains; (ill) the “Fv” fragment consisting of the VL and VH domains of a single antibody; (iv) the “dAb” fragment, which consists of a VH domain; (v) isolated CDR regions; (vi) F(ab')2 fragments, a bivalent fragment comprising two linked Fab fragments; (vii) single chain Fv molecules (scFv), wherein a VH domain and a VL domain are linked by a peptide linker that allows the two domains to associate to form a binding domain; (viii) bi-specific single chain Fv dimers (e.g., U. S. Pat. No. 5,091,513); and (ix) diabodies, multivalent or multispecific fragments constructed by gene fusion (US Patent App. Pub. 2005 / 0214860). Fv, scFv, or diabody molecules may be stabilized by the incorporation of disulphide bridges linking the VH and VL domains. Minibodies comprising a scFv joined to a CH3 domain may also be made (Hu etal., Minibodies are minimized antibody-like proteins comprising a scFv joined to a CH3 domain, Cancer Res. 1996, 56:3055-3061)).

[0269] Murali etal., Antibody like peptidomimetics as large scale immunodetection probes, Cell Mol Biol 2003, 49:209-216, describe a methodology for reducing antibodies into smaller peptidomimetics, they term “antibody like binding peptidomimetics" (ABIP) which may also be useful as an alternative to antibodies.

[0270] " Isolated antibody" or “isolated antibody fragment” refers to the purification status and in such context means the named molecule is substantially free of other biological molecules such as nucleic acids, proteins, lipids, carbohydrates, or other material such as cellular debris and growth media. Generally, the term "isolated" is not intended to refer to a complete absence of such material or to an absence of water, buffers, or salts, unless they are present in amounts that substantially interfere with experimental or therapeutic use of the binding compound as described herein.

[0271] " Monoclonal antibody" or “mAb” or “Mab,” as used herein, refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a multitude of different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are often specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., Continuous cultures of fused cells secreting antibody of predefined specificity, Nature 1975, 256: 495; or may be made by recombinant DNA methods (e.g., U. S. Patent No.

[0272] 4,816,567). The "monoclonal antibodies" may also be isolated from phage antibody libraries using the techniques described in Clackson et a / ., Making antibody fragments using phage display libraries, Nature 1991, 352: 624-628 and Marks et al., By-passing immunization: human antibodies from V-gene libraries displayed on phage, J. Mol. Biol. 1991, 222: 581-597, for example. See also Presta, Selection, design, and engineering oftherapeutic antibodies, J. Allergy Clin. Immunol. 2005,116:731.

[0273] " Chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in an antibody derived from a particular species (e.g., human) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in an antibody derived from another species (e.g., mouse) or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0274] “Human antibody” refers to an antibody that comprises human immunoglobulin protein sequences only. A human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “rat antibody” refer to an antibody that comprises only mouse or rat immunoglobulin sequences, respectively.

[0275] " Humanized antibody" refers to forms of antibodies that contain sequences from non¬ human (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from non-human immunoglobulin, in general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. The prefix “hum,” “hu” or “h” is added to antibody clone designations when necessary to distinguish humanized antibodies from parental rodent antibodies. The humanized forms of rodent antibodies will generally comprise the same CDR sequences of the parental rodent antibodies, although certain amino acid substitutions may be included to increase affinity, increase stability of the humanized antibody, or for other reasons.

[0276] A “variable region” of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. As known in the art, the variable regions of the heavy and light chain each consist of four framework regions (FR) connected by three complementarity determining regions (CDRs) also known as hypervariable regions.

[0277] The term "hypervariable region," " HVR," or " HV" when used herein refers to the regions of an antibody variable domain which are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six HVRs; three in the VH (HI, H2, H3), and three in the VL (LI, L2, L3). In native antibodies, H3 and L3 display the most diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al, Disruption of Early Tumor Necrosis Factor Alpha Signaling Prevents Classical Activation of Dendritic Cells in Lung-Associated Lymph Nodes and Development of Protective Immunity against Cryptococcal Infection, Immunity 2000, 13:37-45; Johnson and Wu, Antibody Engineering Methods and Protocols Methods in Molecular Biology 2003, 248: 1 -25. Indeed, naturally occurring camelid antibodies consisting of a heavy chain only are functional and stable in the absence of light chain. See, e.g., Hamers-Casterman eta / ., Naturally occurring antibodies devoid of light chains, Nature 1993, 363:446-448; Sheriff et al., Similarity between C2 domain jaws and immunoglobulin CDRs, Nature Struct. Biol 1996, 3:733-736. A number of HVR delineations are in use and are encompassed herein. The Kabat Complementarity Determining Regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, 1991). Chothia refers instead to the location of the structural loops (Chothia and Lesk, Canonical structures for the hypervariable regions of immunoglobulins, J. Mol. Biol. 1987, 196:901 -917). The AbM HVRs represent a compromise between the Kabat HVRs and Chothia structural loops, are used by Oxford Molecular's AbM antibody modeling software. The "contact" HVRs are based on an analysis of the available complex crystal structures,

[0278] A “CDR” of a variable domain are amino acid residues within the variable region that are identified in accordance with the definitions of the Kabat, Chothia, the accumulation of both Kabat and Chothia, AbM, contact, and / or conformational definitions or any method of CDR determination well known in the art. Antibody CDRs may be identified as the hypervariable regions originally defined by Kabat et al. See, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, NIH, 1992. The positions of the CDRs may also be identified as the structural loop structures originally described by Chothia and others. See, e.g., Chothia et al., Conformations of immunoglobulin hypervariable regions, Nature, 1989 342:877-883. Other approaches to CDR identification include the “AbM definition,” which is a compromise between Kabat and Chothia and is derived using Oxford Molecular's AbM antibody modeling software (now Accelrys®), or the “contact definition” of CDRs based on observed antigen contacts, set forth in MacCallum et at, Antibody-antigen interactions: contact analysis and binding site topography, J. Mol. Biol., 1996, 262:732-745. In another approach, referred to herein as the “conformational definition” of CDRs, the positions of the CDRs may be identified as the residues that make enthaipic contributions to antigen binding. See, e.g., Makabe ef al., Thermodynamic consequences of mutations in vernier zone residues of a humanized anti-human epidermal growth factor receptor murine antibody, 528, Journal of Biological Chemistry, 2008, 283:1156-1166. Still other CDR boundary definitions may not strictly follow one of the above approaches but will nonetheless overlap with at least a portion of the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. As used herein, a CDR may refer to CDRs defined by any approach known in the art, including combinations of approaches. The methods used herein may utilize CDRs defined according to any of these approaches. For any given embodiment containing more than one CDR, the CDRs may be defined in accordance with any of Kabat, Chothia, extended, AbM, contact, and / or conformational definitions.

[0279] The expression "variable-domain residue-numbering as in Kabat" or "amino-acid-position numbering as in Kabat," and variations thereof, refers to the numbering system used for heavy¬ chain variable domains or light-chain variable domains of the compilation of antibodies in Kabat ef a / ., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FR or HVR of the variable domain. For example, a heavy-chain variable domain may include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc. according to Kabat) after heavy-chain FR residue 82. The Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a "standard" Kabat numbered sequence.

[0280] " Framework" or " FR" residues are those variable-domain residues other than the HVR residues as herein defined.

[0281] A "human consensus framework” or "acceptor human framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences.

[0282] Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, 1991. Examples for the VL, the subgroup may be subgroup kappa I, kappa II, kappa III or kappa IV as in Kabat et al., supra. Additionally, for the VH, the subgroup may be subgroup I, subgroup II, or subgroup III as in Kabat et al., supra. Alternatively, a human consensus framework can be derived from the above in which particular residues, such as when a human framework residue is selected based on its homology to the donor framework by aligning the donor framework sequence with a collection of various human framework sequences. An acceptor human framework "derived from" a human immunoglobulin framework ora human consensus framework may comprise the same amino acid sequence thereof, or it may contain pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes are 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less.

[0283] An "amino-acid modification" at a specified position, e.g., of the Fc region, refers to the substitution or deletion of the specified residue, orthe insertion of at least one amino acid residue adjacent the specified residue. Insertion "adjacent" to a specified residue means insertion within one to two residues thereof. The insertion may be N-terminal or C-terminal to the specified residue. The preferred amino acid modification herein is a substitution.

[0284] " Conservatively modified variants" or "conservative substitution" refers to substitutions of amino acids in a protein with other amino acids having similar characteristics (e.g., charge, side¬ chain size, hydrophobicity / hydrophilicity, backbone conformation and rigidity, etc.), such that the changes can frequently be made without altering the biological activity or other desired property of the protein, such as antigen affinity and / or specificity. Those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (e.g., Watson et al., Molecular Biology of the Gene (4th Ed.), 1987, p. 224). An "affinity-matured" antibody is one with one or more aiterations in one or more HVRs thereof that result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess those aiteration(s). In one embodiment, an affinity-matured antibody has nanomolar or even picomolar affinities for the target antigen. Affinity-matured antibodies are produced by procedures known in the art. For example, Marks ef al., By-passing immunization: Building high affinity human antibodies by chain shuffling, Bio / Technology 1992, 10:779-783, describes affinity maturation by VH- and VL-domain shuffling. Random mutagenesis of HVR and / or framework residues is described by, for example: Barbas et al., In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity, Proc Nat. Acad. Sci. 1994, 91 :3809-3813; Schier et al., Identification of functional and structural amino-acid residues by parsimonious mutagenesis, Gene 1995, 169: 147- 155; Yelton et al., Affinity maturation of the BR96 anti-carcinoma antibody by codon-based mutagenesis, J. Immunol. 1995, 155: 1994-2004; Jackson etat, In vitro antibody maturation. Improvement of a high affinity, neutralizing antibody against IL-1 beta, J. Immunol.

[0285] 1995, 154(7):33 10-9; and Hawkins et al., Selection of phage antibodies by binding affinity: mimicking affinity maturation, J. Moi. Biol. 1992, 226:889-896.

[0286] The term " Fc region" herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy¬ chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. Accordingly, a composition of intact antibodies may comprise antibody populations with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Suitable native-sequence Fc regions for use in the antibodies of the invention include human IgG-1, IgG-2 (IgG2A, IgG2B), IgG-3 and IgG-4.

[0287] " Fc receptor" or " FcR" describes a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of these receptors, FcyRII receptors include FcyRIIA (an "activating receptor") and FcyRIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Activating receptor FcyRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain, inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain, (e.g., M. Daeron, Fc RECEPTOR BIOLOGY, Annu. Rev. Immunol. 1997, 15:203-234. FcRs are reviewed in Ravetch and Kinet, Fc receptors, Annu. Rev. Immunol. 1991, 9: 457-92; Capel etal., Heterogeneity of human IgG Fc receptors, Immunomethods 1994, 4: 25-34; and de Haas et al., Fey receptors of phagocytes, J. Lab. Clin. Med. 1995, 126: 330-41. Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein.

[0288] The term Fc receptor or FcR also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus. Guyer et al., Immunoglobulin binding by mouse intestinal epithelial cell receptors, J. Immunol. 1976, 117: 587, and Tokoyama et al., How do natural killer cells find self to achieve tolerance? Immunity, 1994, 24, 249-257. Methods of measuring binding to FcRn are known (e.g., Ghetie and Ward, FcRn: the MHC class l-related receptor that is more than an IgG transporter, Immunol. Today 1997, 18: (12): 592-8; Ghetie et al., Increasing the serum persistence of an IgG fragment by random mutagenesis, Nat Biotechnol. Jul. 1997;15(7):637-40; Hinton et al., Engineered human IgG antibodies with longer serum half¬ lives in primates, J. Biol. Chem. 2004, 279 (8): 6213-6; WO 2004 / 092219 (Hinton et a!.). Binding to FcRn in vivo and serum half-life of human FcRn high-affinity binding polypeptides can be assayed, e.g., in transgenic mice or transfected human cell lines expressing human FcRn, or in primates to which the polypeptides having a variant Fc region are administered. WO 2004 / 042072 (Presta) describes antibody variants which improved or diminished binding to FcRs. See also, e.g., Shields etai., High Resolution Mapping ofthe Binding Site on Human lgG1 forFcyRI, FcyRI I, FcyRill, and FcRn and Design of lgG1 Variants with Improved Binding to the FcyR, J. Biol. Chem.

[0289] 2001, 9(2): 6591 -6604.

[0290] The phrase "substantially reduced," "substantially different," or “substantially inhibit,” as used herein, denotes a sufficiently high degree of difference between two numeric values (generally one associated with a molecule and the other associated with a reference / comparator molecule) such that one of skill in the art would consider the difference between the two values to be of statistical significance within the context ofthe biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, and / or greater than about 50% as a function ofthe value for the reference / comparator molecule.

[0291] The term "substantially similar" or "substantially the same," as used herein, denotes a sufficiently high degree of similarity between two numeric values (for example, one associated with an antibody ofthe invention and the other associated with a reference / comparator antibody), such that one of skill in the art would consider the difference between the two values to be of little or no biological and / or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and / or less than about 10% as a function of the reference / comparator value.

[0292] As use herein, the term "specifically binds to" or is "specific for" refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody that specifically binds to a target (which can be an epitope) is an antibody that binds this target with greater affinity, avidity, more readily, and / or with greater duration than it binds to other targets. In one embodiment, the extent of binding of an antibody to an unrelated target is less than about 10 percent of the binding of the antibody to the target as measured, e.g., by a radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (Kd) of < 1 pM, < 100 nW, < 10 nW, < 1 nM, or < 0.1 nM. In certain embodiments, an antibody specifically binds to an epitope on a protein that is conserved among the protein from different species. In another embodiment, specific binding can include, but does not require exclusive binding.

[0293] As used herein, the term "immunoadhesin" designates antibody-like molecules which combine the binding specificity of a heterologous protein (an "adhesin") with the effector functions of immunoglobulin constant domains. Structurally, the immunoadhesins comprise a fusion of an amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site of an antibody (i.e., is "heterologous"), and an immunoglobulin constant domain sequence. The adhesin part of an immunoadhesin molecule typically is a contiguous amino acid sequence comprising at least the binding site of a receptor or a ligand. The immunoglobulin constant domain sequence in the immunoadhesin may be obtained from any immunoglobulin, such as IgG-1, IgG-2 (including IgG2A and IgG2B), IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE, IgD or IgM. The Ig fusions preferably include the substitution of a domain of a polypeptide or antibody described herein in the place of at least one variable region within an Ig molecule. In a particularly preferred embodiment, the immunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge, CHI, CH2 and CH3 regions of an lgG-1 molecule. For the production of immunoglobulin fusions, see also US Patent No. 5,428,130 issued June 27, 1995. Immunoadhesin combinations of Ig Fc and ECD of cell surface receptors are sometimes termed soluble receptors.

[0294] A "fusion protein” and a "fusion polypeptide” refer to a polypeptide having two portions covalently linked together, where each of the portions is a polypeptide having a different property. The property may be a biological property, such as activity in vitro or in vivo. The property may also be simple chemical or physical property, such as binding to a target molecule, catalysis of a reaction, etc. The two portions may be linked directly by a single peptide bond or through a peptide linker but are in reading frame with each other.

[0295] A " PD-1 oligopeptide," " PD-L1 oligopeptide," or " PD-L2 oligopeptide" is an oligopeptide that binds, preferably specifically, to a PD-1, PD-L1 or PD-L2 negative costimulatory polypeptide, respectively, including a receptor, ligand or signaling component, respectively, as described herein. Such oligopeptides may be chemically synthesized using known oligopeptide synthesis methodology or may be prepared and purified using recombinant technology. Such oligopeptides are usually at least about 5 amino acids in length, alternatively at least about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 amino acids in length or more. Such oligopeptides may be identified using well known techniques. In this regard, it is noted that techniques for screening oligopeptide libraries for oligopeptides that are capable of specifically binding to a polypeptide target are well known in the art (e.g., U. S. Patent Nos. 5,556,762, 5,750,373, 4,708,871, 4,833,092, 5,223,409, 5,403,484, 5,571,689, 5,663,143; PCT Publication Nos. WO 1984 / 003506 and WO 1984 / 003564; Geysen et al., Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid, Proc. Natl. Acad. Sci. 1984, 81:3998-4002; Geysen ef al., Small peptides induce antibodies with a sequence and structural requirement for binding antigen comparable to antibodies raised against the native protein, Proc. Natl. Acad. Sci. 1985, 82:178-182; Geysen et a!., A priori delineation of a peptide which mimics a discontinuous antigenic determinant, Synthetic Peptides as Antigens, 986, 130-149; Geysen, et al., Strategies for epitope analysis using peptide synthesis, J. Immunol. Meth. 1987, 102, 259- 274; Schoofs etaL Epitopes of an influenza viral peptide recognized by antibody at single amino acid resolution, J. Immunol, 1988, 140:611-616, Cwirla, S. E. et al., Peptides on phage: a vast library of peptides for identifying ligands., Proc. Natl. Acad. Sci. 1990, 87:6378; Lowman, H. B. et al., Selecting high-affinity binding proteins by monovalent phage display, Biochemistry, 1991, 30:10832; Clackson, T. et al., Making antibody fragments using phage display libraries, Nature, 1991, 352: 624; Marks, J. D. et al., By-passing immunization: human antibodies from V-gene libraries displayed on phage, J. Mol. Biol, 1991, 222:581; Kang, et al., Linkage of Recognition and Replication Functions by Assembling Combinatorial Antibody Fab Libraries Along Phage Surfaces, PNAS, 1991, vol. 88, pp. 4363-4366, and Smith, G. P. Surface presentation of protein epitopes using bacteriophage expression systems, Curr. Opin. Biotechnol. 1991, 2:668.

[0296] An "antagonist” antibody or a "blocking" antibody is one that inhibits or reduces a biological activity of the antigen it binds. In some embodiments, blocking antibodies or antagonist antibodies substantially or completely inhibit the biological activity of the antigen. The anti-PD-L1 antibodies of the invention block the signaling through PD-1 so as to restore a functional response by T-cells (e.g., proliferation, cytokine production, target cell killing) from a dysfunctional state to antigen stimulation.

[0297] An "agonist" or “activating antibody” is one that enhances or initiates signaling by the antigen to which it binds. In some embodiments, agonist antibodies cause or activate signaling without the presence of the natural ligand.

[0298] The term "dysfunction" in the context, of immune dysfunction, refers to a state of reduced immune responsiveness to antigenic stimulation. The term includes the common elements of both exhaustion and / or anergy in which antigen recognition may occur, but the ensuing immune response is ineffective to control infection or tumor growth. The term "dysfunctional", as used herein, also includes refractory or unresponsive to antigen recognition, specifically, impaired capacity to translate antigen recognition into down¬ stream T-cell effector functions, such as proliferation, cytokine production and / or target cell killing.

[0299] The term "anergy" refers to the state of unresponsiveness to antigen stimulation resulting from incomplete or insufficient signals delivered through the T-cell receptor (e.g., increase in intracellular Ca+2 in the absence of ras-activation). T cell anergy can also result upon stimulation with antigen in the absence of co- stimulation, resulting in the cell becoming refractory to subsequent activation by the antigen even in the context of co stimulation. The unresponsive state can often be overridden by the presence of lnterleukin-2. Anergic T-cells do not undergo clonal expansion and / or acquire effector functions.

[0300] The term "exhaustion" refers to T cell exhaustion as a state of T cell dysfunction that arises from sustained TCR signaling that occurs during many chronic infections and cancer. It is distinguished from anergy in that it arises not through incomplete or deficient signaling, but from sustained signaling. It is defined by poor effector function, sustained expression of inhibitory receptors and a transcriptional state distinct from that of functional effector or memory T cells. Exhaustion prevents optimal control of infection and tumors. Exhaustion can result from both extrinsic negative regulatory pathways (e.g., immunoregulatory cytokines) as well as cell intrinsic negative regulatory (co stimulatory) pathways.

[0301] " Enhancing T-cell function" means to induce, cause or stimulate a T-cell to have a sustained or amplified biological function, or renew or reactivate exhausted or dysfunctional T-cells. Examples of enhancing T-cell function include: increased secretion of y-interferon from CD4+ or CD8+ T-cells, increased proliferation, increased survival, increased differentiation, increased antigen responsiveness (e.g., viral, pathogen, or tumor clearance) relative to such levels before the intervention. In some embodiments, the level of enhancement is as least 50%, alternatively 60%, 70%, 80%, 90%, 100%, 120%, 150%, 200%. The manner of measuring this enhancement is known to one of ordinary skill in the art.

[0302] As used herein, "metastasis" or “metastatic” is meant the spread of cancer from its primary site to other places in the body. Cancer cells can break away from a primary tumor, penetrate into lymphatic and blood vessels, circulate through the bloodstream, and grow in a distant focus (metastasize) in normal tissues elsewhere in the body. Metastasis can be local or distant. Metastasis is a sequential process, contingent on tumor cells breaking off from the primary tumor, traveling through the bloodstream, and stopping at a distant site. At the new site, the cells establish a blood supply and can grow to form a life-threatening mass. Both stimulatory and inhibitory molecular pathways within the tumor cell regulate this behavior, and interactions between the tumor cell and host cells in the distant site are also significant.

[0303] The term “cancer,” “cancerous,” or “malignant” refers to or describe the physiological condition in subjects that is typically characterized by unregulated cell growth. The term “cancer” includes but is not limited to a primary cancer that originates at a specific site in the body, a metastatic cancer that has spread from the piace in which it started to other parts of the body, a recurrence from the original primary cancer after remission, and a second primary cancerthat is a new primary cancer in a person with a history of previous cancer of a different type from the latter one. Examples of cancer include, but are not limited to, brain cancer, head / neck cancer (including squamous cell carcinoma of the head and neck (SCCHN)), prostate cancer, ovarian cancer, bladder cancer (including urothelial carcinoma, also known as transitional cell carcinoma (TCC)), lung cancer (including squamous cell carcinoma, small cell lung cancer (SCLC), and non- smail cell lung cancer (NSCLC)), breast cancer, bone cancer, colorectal cancer, kidney cancer, liver cancer (including hepatocellular carcinoma (HCC)), stomach cancer, pancreatic cancer, esophageal cancer, cervical cancer, sarcoma, skin cancer (including melanoma and Merkel cell carcinoma (MCC)), multiple myeloma, mesothelioma, malignant rhabdoid tumors, diffuse intrinsic pontine glioma (DIPG), carcinoma, lymphoma, diffuse large B-cell lymphoma (DLBCL), primary mediastinal B-cell lymphoma (PMBCL), follicular lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CIVIL), follicular lymphoma, Hodgkin’s lymphoma (HL), classical Hodgkin lymphoma (cHL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid cell leukemia-1 protein (Mcl-1), myelodysplastic syndrome (MDS), non-Hodgkin’s lymphoma (NHL), small lymphocytic lymphoma (SLL), and SWI / SNF-mutant cancer.

[0304] As used herein, “breast cancer” includes, but is not limited to, luminal A, luminal B, triple negative / basal-like, or HER2-enriched subtypes. Breast cancers may be estrogen receptor (ER)-positive and / or progesterone receptor (PR)-positive, alternatively referred to as hormone receptor (HR)-positive. HR-positive breast cancers may be human epidermal growth factor receptor 2 (HER2)-negative (i.e., HR+ / HER2-) or HER2-positive ( / .©., HR+ / HER2+). HR-negative breast cancers may be HER2-positive ( / .e., HR- / HER2+) or HER-negative (HR- / HER2-), i.e. “triple negative” breast cancer (TNBC). In some embodiments, the breast cancer demonstrates primary or acquired resistance to endocrine therapy, anti-HER2 agents and / or CDK4 / CDK6 inhibitors. In some embodiments, the breast cancer is advanced or metastatic breast cancer. In some embodiments of the foregoing, the breast cancer is characterized by amplification or overexpression of CCNEI and / or CCNE2.

[0305] As used herein, “in combination with” or "in conjunction with" refers to administration of one treatment modality in addition to at least one other treatment modality. As such, “in combination with” or "in conjunction with" refers to administration of one treatment modality before, during, or after administration of at least one other treatment modality to the individual.

[0306] An "objective response" refers to a measurable response, including complete response (CR) or partial response (PR). In some embodiments, the term "objective response rate" (ORR) refers to the sum of complete response (CR) rate and partial response (PR) rate. " Complete response" or " CR," as used herein, means the disappearance of all signs of cancer (e.g., disappearance of all target lesions) in response to treatment. This does not always mean the cancer has been cured.

[0307] As used herein, "partial response" or " PR" refers to a decrease in the size of one or more tumors or lesions, or in the extent of cancer in the body, in response to treatment. For example, in some embodiments, PR refers to at least a 30% decrease in the sum of the longest diameters (SLD) of target lesions, taking as reference the baseline SLD.

[0308] As used herein, "progressive disease" or " PD" refers to at least a 20% increase in the SLD of target lesions, taking as reference the smallest SLD recorded since the treatment started or the presence of one or more new lesions.

[0309] As used herein, "progression free survival" or“PFS” refers to the length of time during and after treatment during which the disease being treated (e.g., cancer) does not get worse. Progression-free survival may include the amount of time patients have experienced a complete response or a partial response, as well as the amount of time patients have experienced stable disease.

[0310] As used herein, "overall response rate" (ORR) refers to the sum of complete response (CR) rate and partial response (PR) rate.

[0311] As used herein, "overall survival" refers to the percentage of individuals in a group who are likely to be alive after a particular duration of time.

[0312] " Sustained response" refers to the sustained effect on reducing tumor growth after cessation of a treatment. For example, the tumor size may be the same size or smaller as compared to the size at the beginning of the medicament administration phase. In some embodiments, the sustained response has a duration of at least the same as the treatment duration, at least 1,5x, 2x, 2.5x, or 3x length of the treatment duration, or longer.

[0313] “Duration of Response’’ for purposes of the present invention means the time from documentation of tumor model growth inhibition due to drug treatment to the time of acquisition of a restored growth rate similar to pretreatment growth rate.

[0314] In some embodiments, the anti-cancer effect of the method of treating cancer, including “objective response,” “complete response,” “partial response,” “progressive disease,” “stable disease,” “progression free survival,” “duration of response,” as used herein, are as defined and assessed by the investigators using RECIST v1.1 (Eisenhauer et al., New response evaluation criteria in solid tumors: revised RECIST guideline, Eur J of Cancer 2009; 45(2):228-47) in patients with locally advanced or metastatic solid tumors other than metastatic CRPC, and RECIST v1.1 and PCWG3 (Scher et al., Trial Design and Objectives for Castration-Resistant Prostate Cancer: Updated Recommendations From the Prostate Cancer Clinical Trials Working Group 3, J Clin Oncol 2016; 34(12): 1402-18) in patients with metastatic CRPC. The disclosures of Eisenhauer et al., 2009and Scher et al., 2016 are herein incorporated by references in their entireties. The term “patient” or “subject” refers to any subject for which therapy is desired or that is participating in a clinical trial, epidemiological study or used as a control, including humans and non-human animals, including veterinary subjects such as cattle, horses, dogs and cats. In a preferred embodiment, the subject is a human and may be referred to as a patient. Those skilled in the medical art are readily able to identify individual patients who are afflicted with cancer.

[0315] In some embodiments, the combination or co-administration of two or more agents can be useful for treating individuals suffering from cancer who have primary or acquired resistance to ongoing therapies. The combination therapy provided herein may be useful for improving the efficacy and / or reducing the side effects of cancer therapies for individuals who do respond to such therapies.

[0316] As used herein, the term “combination therapy” refers to the administration of each agent of the combination therapy of the invention, either alone or in a medicament, either simultaneously, separately or sequentially, as mixed or individual dosages.

[0317] As used herein, the term “simultaneously,” "simultaneous administration,” "administered simultaneously,” “concurrently,” or "concurrent administration,” means that the agents are administered at the same point in time or immediately following one another, but that the agents can be administered in any order. For example, in the latter case, the two or more agents are administered at times sufficiently close that the results observed are indistinguishable from those achieved when the agents are administered at the same point in time. The term simultaneous includes the administration of each agent of the combination therapy of the invention in the same medicament.

[0318] The agents of the present invention can be administered completely separately or in the form of one or more separate compositions. For example, the agents may be given separately at different times during the course of therapy (in a chronologically staggered manner, especially a sequence-specific manner) in such time intervals that the combination therapy is effective in treating cancer.

[0319] As used herein, the term “sequential,” “sequentially,” “administered sequentially,” or “sequential administration” refers to the administration of each agent of the combination therapy of the invention, either alone or in a medicament, one after the other, wherein each agent can be administered in any order. Sequential administration may be particularly useful when the therapeutic agents in the combination therapy are in different dosage forms, for example, one agent is a tablet and another agent is a sterile liquid, and / or the agents are administered according to different dosing schedules, for example, one agent is administered daily, and the second agent is administered less frequently such as weekly.

[0320] As used herein, “in combination with,” "in conjunction with" or “combined administration” refers to administration of one agent in addition to at least one other agent. As such, “in combination with,” ’’in conjunction with” or “combined administration” refers to administration of one agent before, during, or after administration of at least one other agent to the individual. The administration of two or more agents are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

[0321] A “combination” or “pharmaceutical combination” refers to a combination of any two or more agents as described herein. These two or more agents may (but do not necessarily) belong to different classes of agents.

[0322] In some embodiments, a combination as described herein, e.g., a CDK4 inhibitor in combination with a PD-1 axis binding antagonist, is administered in a single dose, in some embodiments, a combination as described herein, e.g., a CDK4 inhibitor in combination with a PD-1 axis binding antagonist, is administered in multiple doses. In some embodiments, an amount of a combination as described herein, e.g., a CDK4 inhibitor in combination with a PD-1 axis binding antagonist, may be administered periodically at regular intervals (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times every 1, 2, 3, 4, 5, or 6 days, or every 1, 2, 3, 4, 5, 6, 7, 8, or 9 weeks, or every 1, 2, 3, 4, 5, 6, 7, 8, 9 months or longer).

[0323] In some embodiments, a combination as described herein, e.g., a CDK4 inhibitor in combination with a PD-1 axis binding antagonist, is administered at a predetermined interval (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times every 1, 2, 3, 4, 5, or 6 days, or every 1, 2, 3, 4, 5, 6, 7, 8, or 9 weeks, or every 1, 2, 3, 4, 5, 6, 7, 8, 9 months or longer).

[0324] The present invention relates to combinations of two or more agents for simultaneous, separate or sequential administration. For example, the individual agents of the combination of the invention can be administered separately at different times in any order during the course of therapy or concurrently in divided or single combination forms.

[0325] The terms "concurrent administration,” "administration in combination," "simultaneous administration" or "administered simultaneously," as used herein, means that the agents are administered at the same point in time or immediately following one another. For example, in the latter case, the two agents are administered at times sufficiently close that the results observed are indistinguishable from those achieved when the agents are administered at the same point in time.

[0326] The agents of the present invention can be administered completely separately or in the form of one or more separate compositions. For example, the agents may be given separately at different times during the course of therapy (in a chronologically staggered manner, especially a sequence-specific manner) in such time intervals that the combination therapy is effective in treating cancer.

[0327] The term “sequentially,” as used herein, refers to a treatment in which administration of a first treatment, such as administration of first agent, follows administration of a second treatment, such as administration of a second agent.

[0328] The dosage of the individual agents of the combination may require more frequent administration of one of the agent(s) as compared to the other agent(s) in the combination. Therefore, to permit appropriate dosing, packaged pharmaceutical products may contain one or more dosage forms that contain the combination of agents, and one or more dosage forms that contain one of the combination of agents, but not the other agent(s) of the combination.

[0329] The term “single formulation,” as used herein, refers to a single carrier or vehicle formulated to deliver effective amounts of both therapeutic agents to a subject. The single vehicle is designed to deliver an effective amount of each of the agents, along with any pharmaceutically acceptable carriers or excipients. In some embodiments, the vehicle is a tablet, capsule, pill, or a patch. In other embodiments, the vehicle is a solution or a suspension.

[0330] The term “unit dose” is used herein to mean simultaneous administration of both agents together, in one dosage form, to the subject being treated. In some embodiments, the unit dose is a single formulation. In certain embodiments, the unit dose includes one or more vehicles such that each vehicle includes an effective amount of at least one of the agents along with pharmaceutically acceptable carriers and excipients. In some embodiments, the unit dose is one or more tablets, capsules, pills, or patches administered to the subject at the same time.

[0331] An “oral dosage form” includes a unit dosage form prescribed or intended for oral a ministration.

[0332] The term “advanced,” as used herein, as it relates to breast cancer, includes locally advanced (non-metastatic) disease and metastatic disease.

[0333] The term “treat” or “treating” a cancer, as used herein means to administer a combination therapy according to the present invention to a subject having cancer, or diagnosed with cancer, to achieve at least one positive therapeutic effect, such as, for example, reduced number of cancer cells, reduced tumor size, reduced rate of cancer cell infiltration into peripheral organize, or reduced rate of tumor metastases or tumor growth, reversing, stopping, controlling, slowing, interrupting, arresting, alleviating, and / or inhibiting the progression or severity of a sign, symptom, disorder, condition, or disease, but does not necessarily involve a total elimination of all disease-related signs, symptoms, conditions, or disorders. Within the meaning of the present invention, the term “treat” or “treating” also denotes, to arrest, delay the onset (i.e., the period prior to clinical manifestation of a disease or symptom of a disease) and / or reduce the risk of developing or worsening a symptom of a disease.

[0334] The term "treatment," as used herein, unless otherwise indicated, refers to the act of treating as "treating" is defined immediately above. The term “treating” also includes adjuvant and neo-adjuvant treatment of a subject. For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: reducing the proliferation of (or destroying) neoplastic or cancerous cell; inhibiting metastasis or neoplastic cells; shrinking or decreasing the size of tumor; remission of the cancer; decreasing at least one symptom resulting from the cancer; increasing the quality of life of those suffering from the cancer; decreasing the dose of other medications required to treat the cancer; delaying the progression the cancer; curing the cancer; overcoming one or more resistance mechanisms of the cancer; and / or prolonging survival of patients with cancer. Positive therapeutic effects in cancer can be measured in a number of ways (e.g., W. A. Weber, J. Nucl. Med. 50:1S-10S (200)). In some embodiments, the treatment achieved by a combination of the invention is any of the partial response (PR), complete response (CR), overall response (OR), progression free survival (PFS), disease free survival (DFS) and overall survival (OS). PFS, also referred to as “Time to Tumor Progression" indicates the length of time during and after treatment that the cancer does not grow and includes the amount of time patients have experienced a CR or PR, as well as the amount of time patients have experienced stable disease (SD). DFS refers to the length of time during and after treatment that the patient remains free of disease. OS refers to a prolongation in life expectancy as compared to naive or untreated subjects or patients. In some embodiments, response to a combination of the invention is any of PR, CR, OR, OS, PFS, or DFS that is assessed using Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 response criteria. The treatment regimen for a combination of the invention that is effective to treat a cancer patient may vary according to factors such as the disease state, age, weight of the patient, and the ability of the therapy to elicit an anti-cancer response in the subject. While an embodiment of any of the aspects of the invention may not be effective in achieving a positive therapeutic effect in every subject, it should do so in a statistically significant number of subjects as determined by any statistical test known in the art such as the Student’s t-test, the chi2-test the U-test according to Mann and Whitney, the Kruskal-Wallis test (H-test), Jonckheere-Terpstrat-testy and the Wilcon on-test.

[0335] The term “administer," “administering,” or “administration,” “treat,” “treating,” or “treatment” as it applies to an animal, human, experimental subject, cell, tissue, organ or biological fluid, refers to contacting, implanting, absorbing, ingesting, injecting, inhaling, or introducing of an exogenous pharmaceutical, therapeutic or diagnostic agent, compound, particle, and / or composition, to the animal, human, experimental subject, cell, tissue, organ or biological fluid. Treatment of a cell encompasses contact of an agent to the cell, as well as contact of an agent to a fluid, where the fluid is in contact with the cell. The term “treatment" also encompasses in vitro and ex vivo treatment, e.g., of a cell, by a reagent, diagnostic, binding compound, or by another cell.

[0336] The term "diagnosis" is used herein to refer to the identification or classification of a molecular or pathological state, disease or condition (e.g.. cancer). For example, "diagnosis" may refer to identification of a particular type of cancer. " Diagnosis" may also refer to the classification of a particular subtype of cancer, e.g., by histopathological criteria, or by molecular features (e.g., a subtype characterized by expression of one or a combination of biomarkers (e.g., particular genes or proteins encoded by said genes)).

[0337] The term "aiding diagnosis" is used herein to refer to methods that assist in making a clinical determination regarding the presence, or nature, of a particular type of symptom or condition of a disease or disorder (e.g., cancer). For example, a method of aiding diagnosis of a disease or condition (e.g., cancer) can comprise measuring certain biomarkers in a biological sample from an individual

[0338] The term "sampie," as used herein, refers to a composition that is obtained or derived from a subject and / or individual of interest that contains a cellular and / or other molecular entity that is to be characterized and / or identified, for example based on physical, biochemical, chemical and / or physiological characteristics. For example, the phrase "disease sample" and variations thereof refers to any sample obtained from a subject of interest that would be expected or is known to contain the cellular and / or molecular entity that is to be characterized. Samples include, but are not limited to, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous fluid, lymph fluid, synovial fluid, follicular fluid, seminal fluid, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebro-spinal fluid, saliva, sputum, tears, perspiration, mucus, tumor lysates, and tissue culture medium, tissue extracts such as homogenized tissue, tumor tissue, cellular extracts, and combinations thereof.

[0339] By "tissue sample" or "cell sample" is meant a collection of similar cells obtained from a tissue of a subject or individual. The source of the tissue or cell sample may be solid tissue as from a fresh, frozen and / or preserved organ, tissue sample, biopsy, and / or aspirate; blood or any blood constituents such as plasma; bodily fluids such as cerebral spinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid: cells from any time in gestation or development of the subject. The tissue sample may also be primary or cultured cells or cell lines. Optionally, the tissue or cell sample is obtained from a disease tissue / organ, The tissue sample may contain compounds which are not naturally intermixed with the tissue in nature such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, or the like.

[0340] A "reference sample,” "reference cell,” "reference tissue,” "control sample,” "control cell,” or "control tissue,” as used herein, refers to a sample, cell, tissue, standard, or level that is used for comparison purposes. In one embodiment, a reference sample, reference ceil, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non¬ diseased part of the body (e.g., tissue or cells) of the same subject or individual. For example, healthy and / or non-diseased cells or tissue adjacent to the diseased cells or tissue e.g., cells or tissue adjacent to a tumor). In another embodiment, a reference sample is obtained from an untreated tissue and / or cell of the body of the same subject or individual. In yet another embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-diseased part of the body (e.g., tissues or cells) of an individual who is not the subject or individual. In even another embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from an untreated tissue and / or cell of the body of an individual who is not the subject or individual.

[0341] The term “pharmaceutical composition” refers to a preparation which is in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. Such formulations are sterile. “Pharmaceutically acceptable” carriers or excipients (vehicles, additives) are those which can reasonably be administered to a subject to provide an effective dose of the active ingredient employed.

[0342] The combinations provided herein may be formulated by a variety of methods apparent to those of skill in the art of pharmaceutical formulation. The various release properties described above may be achieved in a variety of different ways. Suitable formulations include, for example, tablets, capsules, press coat formulations, and other easily administered formulations.

[0343] A "package insert" refers to instructions customarily included in commercial packages of medicaments that contain information about the indications customarily included in commercial packages of medicaments that contain information about the indications, usage, dosage, administration, contraindications, other medicaments to be combined with the packaged product, and / or warnings concerning the use of such medicaments, etc.

[0344] An “amount” for use and for treating a subject refers to an amount that provides, in single or multiple doses, alone, or in combination with one or more other agents, a detectable response of any duration of time (transient, medium or long term), a desired outcome in or an objective or subjective benefit to a subject of any measurable or detectable degree or for any duration of time (e.g., for hours, days, months, years, in remission or cured). Such amounts typically are effective to ameliorate a disease, or one, multiple or all adverse effects / symptoms, consequences or complications of the disease, to a measurable extent, although reducing or inhibiting a progression or worsening of the disease, or providing stability (i.e., not worsening) state of the disease, is considered a satisfactory outcome. The term “therapeutically effective amount” also means an amount of an agent, alone, or in combination with one or more other agents, effective for producing a desired therapeutic effect upon administration to a subject, for example, to stem the growth, or result in the shrinkage, of a cancerous tumor. In reference to the treatment of cancer, a therapeutically effective amount refers to that amount which has the effect of (1) reducing the size of the tumor, (2) inhibiting (that is, slowing to some extent, preferably stopping) tumor metastasis emergence, (3) inhibiting to some extent (that is, slowing to some extent, preferably stopping) tumor growth or tumor invasiveness, and / or (4) relieving to some extent (or, preferably, eliminating) one or more signs or symptoms associated with the cancer. Therapeutic or pharmacological effectiveness of the doses and administration regimens may also be characterized as the ability to induce, enhance, maintain or prolong disease control and / or overall survival in patients with these specific tumors, which may be measured as prolongation of the time before disease progression,

[0345] A therapeutically effective amount can be administered in one or more administrations. For the purposes of this invention, a therapeutically effective amount is an amount sufficient to accomplish prophylactic or therapeutic treatment either directly or indirectly. As is understood in the clinical context, an effective amount of a drug, compound or pharmaceutical composition may or may not be achieved in conjunction with another drug, agent, component, composition, compound, substance, targeted agent, targeted therapeutic agent, therapeutic antibody, therapeutic agent, medicament or pharmaceutical composition.

[0346] As used herein, “ameliorate” refers to any reduction in the extent, severity, frequency, and / or likelihood of a symptom or clinical sign characteristic of a particular disease. “Symptom” refers to any subjective evidence of disease or of a subject's condition.

[0347] Administration of the compounds of the present invention may be effected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical, and rectal administration. Each compound may be administered according to the same or different route of administration.

[0348] The term "biosimilar" refers to a biological product that is highly similar to an FDA- approved biological product (reference product) and has no clinically meaningful differences in terms of pharmacokinetics, safety and efficacy from the reference product.

[0349] The term "bioequivalent" refers to a biological product that is pharmaceutically 5 equivalent and has a similar bioavailability to an FDA-approved biological product (reference product). For example, according to the FDA the term bioequivalence is defined as "the absence of a significant difference in the rate and extent to which the active ingredient or active moiety in pharmaceutical equivalents or pharmaceutical alternatives becomes available at the site of drug action when administered at the same molar dose under similar conditions 10 in an appropriately designed study" (United States Food and Drug Administration, " Guidance for Industry: Bioavailability and Bioequicalence Studies for Orally Administered Drug Products - General Considerations," 2003, Center for Drug Evaluation and Research),

[0350] The term "biobetter" refers a biological product that is in the same class as an FDA approved biological product (reference product) but is not identical and is improved in terms 15 of safety, efficacy, stability, etc. over the reference product.

[0351] “Tumor” as it applies to a subject diagnosed with, or suspected of having, a 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 usually does not contain cysts or liquid areas. Examples of solid tumors are sarcomas, carcinomas, and lymphomas.

[0352] The term “tumor size” refers to the total size of the tumor which can be measured as the length and width of a tumor. Tumor size may be determined by a variety of methods known in the art, such as, e.g., by measuring the dimensions of tumor(s) upon removal from the subject, e.g., using calipers, or while in the body using imaging techniques, e.g., bone scan, ultrasound, CR or MRI scans.

[0353] The term “additive” is used to mean that the result of the combination of two or more agents is no greater than the sum of each agent individually. In one embodiment, the combination of agents described herein displays a synergistic effect. The term “synergy” or “synergistic” are used to mean that the result of the combination of two or more agents is greater than the sum of each agent individually. This improvement in the disease, condition or disorder being treated is a “synergistic” effect. A “synergistic amount” is an amount of the combination of the two or more agents that results in a synergistic effect, as “synergistic” is defined herein. A “synergistic combination” refers to a combination of agents which produces a synergistic effect in vivo, or alternatively in vitro as measured according to the methods described herein.

[0354] Determining a synergistic interaction between two or more agents, the optimum range for the effect and absolute dose ranges of each agent for the effect may be definitively measured by administration of the agents over different dose ranges, and / or dose ratios to subjects in need of treatment. However, the observation of synergy in in vitro models or in vivo models can be predictive of the effect in humans and other species and in vitro models or / n vivo models exist, as described herein, to measure a synergistic effect. The results of such studies can also be used to predict effective dose and plasma concentration ratio ranges and the absolute doses and plasma concentrations required in humans and other species such as by the application of pharmacokinetic and / or pharmacodynamics methods.

[0355] The term "pharmaceutically acceptable salt," as used herein, refers to pharmaceutically acceptable organic or inorganic salts of a compound of the invention. Some embodiments also relate to the pharmaceutically acceptable acid addition salts of the compounds described herein. Suitable acid addition salts are formed from acids which form non-toxic salts. Non-limiting examples of suitable acid addition salts, i.e., salts containing pharmacologically acceptable anions, include, but are not limited to, the acetate, acid citrate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulphate / sulphate, bitartrate, borate, camsylate, citrate, cyclamate, edisylate, esylate, ethanesulfonate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyrogiutamate, saccharate, stearate, succinate, tannate, tartrate, p-toluenesulfonate, trifluoroacetate and xinofoate salts.

[0356] Additional embodiments relate to base addition salts of the compounds described herein. Suitable base addition salts are formed from bases which form non-toxic salts. Non-limiting examples of suitable base salts include the aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts.

[0357] The compounds described herein that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that may be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds described herein are those that form non-toxic acid addition sails, e.g., salts containing pharmacologically acceptable anions, such as the hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate.e., 1,1’-methylene-bis-(2-hydroxy-3-naphthoate)] salts. The compounds described herein that include a basic moiety, such as an amino group, may form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above.

[0358] The chemical bases that may be used as reagents to prepare pharmaceutically acceptable base salts of those compounds described herein that are acidic in nature are those that form non-toxic base salts with such compounds. Such non-toxic base salts include but are not limited to those derived from such pharmacologically acceptable cations such 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 the lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines. Hemisalts of acids and bases may also be formed, for example, hemisulphate and hemicalcium salts.

[0359] For a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for making pharmaceutically acceptable salts of compounds described herein are known to one of skill in the art.

[0360] " Carriers," as used herein, include pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to the cell or subject being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0361] The term “solvate” is used herein to describe a molecular complex comprising a compound described herein and one or more pharmaceutically acceptable solvent molecules, for example, water and ethanol. The compounds described herein may also exist in unsolvated and solvated forms. Accordingly, some embodiments relate to the hydrates and solvates of the compounds described herein.

[0362] Compounds described herein containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. Where a compound described herein contains an alkenyl or alkenylene group, geometric cis / trans (or Z / E) isomers are possible. Where structural isomers are interconvertible via a low energy barrier, tautomeric isomerism (‘tautomerism’) can occur. This can take the form of proton tautomerism in compounds described herein containing, for example, an imino, keto, or oxime group, or so-called valence tautomerism in compounds which contain an aromatic moiety, A single compound may exhibit more than one type of isomerism.

[0363] The compounds of the embodiments described herein include all stereoisomers (e.g., cis and trans isomers) and all optical isomers of compounds described herein (e.g., R and S enantiomers), as well as racemic, diastereomeric and other mixtures of such isomers. While all stereoisomers are encompassed within the scope of our claims, one skilled in the art will recognize that particular stereoisomers may be preferred.

[0364] METHODS, USES AND MEDICAMENTS

[0365] General Methods

[0366] Standard methods in molecular biology are described in Sambrook, Fritsch and Maniatis (1982 & 19892nd Edition, 2001 3rd Edition) Molecular Cloning, A Laboratory Manual; Sambrook and Russell Molecular Cloning, 3rd ed., 2001; Wu, Recombinant DNA, Vol. 217. Standard methods also appear in Ausbel, et al., Current Protocols in Molecular Biology, Vols.1-4, 2001, which describes cloning in bacterial cells and DNA mutagenesis (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3), and bioinformatics (Vol.

[0367] 4).

[0368] Methods for protein purification including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization are described (Coligan, et al., Current Protocols in Protein Science, Vol. 1, 2000, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and glycosylation of proteins are described (e.g., Coligan, etal., Current Protocols in Protein Science, Vol. 2, 2000; Ausubel, et al., Current Protocols in Molecular Biology, Vol. 3, 2001, pp. 16.0.5- 16.22.17; Sigma-Aldrich, Co. Products for Life Science Research, 2001, pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, pp. 384-391). Production, purification, and fragmentation of polyclonal and monoclonal antibodies are described (Coligan, et at, Current Protocols in Immunology, Vol. 1, 2001; Harlow and Lane, Using Antibodies, 1999). Standard techniques for characterizing ligand / receptor interactions are available (e.g., Coligan, et al., Current Protocols in Immunology, Vol. 4, 2001). Monoclonal, polyclonal, and humanized antibodies can be prepared (e.g., Sheperd and Dean (eds.) Monoclonal Antibodies, 2000; Kontermann and Dubel (eds.) Antibody Engineering, 2001; Harlow and Lane, Antibodies A Laboratory Manual, 1988, pp. 139-243; Carpenter, et al., Non-Fc receptor-binding humanized anti-CD3 antibodies induce apoptosis of activated human T cells, J. Immunol. 2000, 165:6205; He, et al., Humanization and pharmacokinetics of a monoclonal antibody with specificity for both E-and P-selectin, J. Immunol. 1998, 160:1029; Tang et al., Use of a peptide mimotope to guide the humanization of MRK-16, an anti-P-glycoprotein monoclonal antibody, J. Biol. Chem. 1999, 274:27371-27378; Baca ef al., Antibody humanization using monovalent phage display, J, Biol. Chem. 1997, 272:10678-10684; Chothia etal., Conformations of immunoglobulin hypervariable regions, Nature 1989, 342:877-883; Foote and Winter Antibody framework residues affecting the conformation of the hypervariable loops, J. Mol. Biol. 1992, 224:487-499; U. S. Patent No. 6,329,511).

[0369] An alternative to humanization is to use human antibody libraries displayed on phage or human antibody libraries in transgenic mice (Vaughan et al., Human antibodies with sub- nanomolar affinities isolated from a large non-immunized phage display library, Nature Biotechnol. 1996, 14:309-314; Barbas, Synthetic human antibodies, Nature Medicine 1995, 1:837-839; Mendez et al., Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response in mice, Nature Genetics 1997, 15:146-156; Hoogenboom and Chames, Natural and designer binding sites made by phage display technology, Immunol. Today 2000, 21:371-377; Barbas et al., Phage Display: A Laboratory Manual, 2001; Kay et al., Phage Display of Peptides and Proteins: A Laboratory Manual, 1996; de Bruin et al., Selection of high-affinity phage antibodies from phage display libraries, Nature Biotechnol. 1999, 17:397-399).

[0370] Purification of antigen is not necessary for the generation of antibodies. Animals can be immunized with cells bearing the antigen of interest. Splenocytes can then be isolated from the immunized animals, and the splenocytes can be fused with a myeloma cell line to produce a hybridoma (e.g., Meyaard, L., et. al., LAIR-1, a novel inhibitory receptor expressed on human mononuclear leukocytes, Immunity 1997, 7:283-290; Wright etal., Inhibition of chicken adipocyte differentiation by in vitro exposure to monoclonal antibodies against embryonic chicken adipocyte plasma membranes, immunity 2000, 13:233-242; Preston, et al., The leukocyte / neuron cell surface antigen 0X2 binds to a ligand on macrophages,) Eur. J. Immunol. 1997, 27:1911-1918, Kaithamana et al., Induction of experimental autoimmune Graves' disease in BALB / c mice, J. Immunol. 1999, 163:5157-5164).

[0371] Antibodies can be conjugated, e.g., to small drug molecules, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic, kit or other purposes, and include antibodies coupled, e.g., to dyes, radioisotopes, enzymes, or metals, e.g., colloidal gold (e.g., Le Doussal et al., Enhanced in wvo targeting of an asymmetric bivalent hapten to double-antigen-positive mouse B cells with monoclonal antibody conjugate cocktails, J. Immunol. 1991, 146:169-175; Gibellini et al., Extracellular HIV-1 Tat protein induces the rapid Ser133 phosphorylation and activation of CREB transcription factor in both Jurkat iymphoblastoid T cells and primary..., J. Immunol. 1998160:3891-3898; Hsing and Bishop, Requirement for nuclear factor-xB activation by a distinct subset of CD40-mediated effector functions in B lymphocytes, J. Immunol. 1999, 162:2804-2811; Everts et al., Selective intracellular delivery of dexamethasone into activated endothelial cells using an E-selectin-directed immunoconjugate, J. Immunol. 2002, 168:883-889).

[0372] Methods for flow cytometry, including fluorescence activated cell sorting (FACS), are available (e.g., Owens, et al., Flow Cytometry Principles for Clinical Laboratory Practice, 1994; Givan Flow Cytometry, 2nd ed.; 2001; Shapiro, Practical Flow Cytometry, 2003). Fluorescent reagents suitable for modifying nucleic acids, including nucleic acid primers and probes, polypeptides, and antibodies, for use, e.g., as diagnostic reagents, are available (Molecular Probes, Catalogue, 2003; Sigma-Aldrich, Catalogue, 2003.

[0373] Standard methods of histology of the immune system are described (e.g., Muller- Harmelink (ed.), Human Thymus: Histopathology and Pathology, 1986; Hiatt, et al., Color Atlas of Histology, 2000; Louis, etai., Basic Histology: Text and Atlas, 2002.

[0374] Software packages and databases for determining, e.g., antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments, are available (e.g., GenBank, Vector NTI® Suite (Informax, Inc, Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DeCypher® (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al., A comparison of signal sequence prediction methods using a test set of signal peptides, Bioinformatics 2000, 16: 741-742; Menne, K. M. L., et. ai. A comparisonof signal sequence prediction methods using a test set of signal peptides, Bioinformatics 2000, 16, 741-742; Wren, et al., SIGNAL-sequence information and GeNomic Ana Lysis Comput. Methods Programs Biomed. 2002, 68:177-181; von Heijne, Patterns of amino acids near signal-sequence cleavage sites, Eur. J. Biochem. 1983, 133:17-21; von Heijne, A new method for predicting signal sequence cleavage sites, Nucleic Acids Res. 1986, 14:4683-4690).

[0375] In one aspect, the antibody against PD-L1, and / or PD-1 may incorporated into a multi¬ specific antibody (e.g., a bispecific antibody). In some such embodiments, a bispecific antibody comprises a first antibody variable domain and a second antibody variable domain, wherein the first antibody variable domain is capable of recruiting the activity of a human immune effector cell by specifically binding to an effector antigen located on the human immune effector cell, and wherein the second antibody variable domain is capable of specifically binding to a target antigen as provided herein. In some embodiments, the antibody has an lgG1, lgG2, lgG3, or lgG4 isotype. In some embodiments, the antibody comprises an immunologically inert Fc region. In some embodiments the antibody is a human antibody or humanized antibody.

[0376] in some embodiments, the bispecific antibody provided herein binds to two different target antigens on the same target ceil (e.g., two different antigens on the same tumor cell). Such antibodies may be advantageous, for example, for having increased specificity for a target ceil of interest (e.g., for a tumor cell that expresses two particular tumor associated antigens of interest). For example, In some embodiments, a bispecific antibody provided herein comprises a first antibody variable domain and a second antibody variable domain, wherein the first antibody variable domain is capable of specifically binding to a first target antigen as provided herein and the second antibody variable domain is capable of specifically binding to a second target antigen as provided herein. In some embodiments, the first target antigen is PD-L1 and the second target antigen is CD47. Examples of mAbs that bind to human PD-L1 and that may be used in bispecific anti-PD-L.1 / anti-CD47 antibodies include antibodies described in WO 2013 / 079174, WO 2015 / 061668, WO 2010 / 089411, WO 2007 / 005874, WO 2010 / 036959, WO 2014 / 100079, WO 2013 / 019906, WO 2010 / 077634, and U. S. Patent Nos. 8,552,154, 8779,108, and 8,383,796. Examples of mAbs that bind to CD47 and that may be used in bispecific anti-PD-LI / anti-CD47 antibodies include the anti-CD47 antibodies Hu5F9-G4 (Forty Seven Inc.), CC-90002 (Celgene), SRF231, and B6H12.

[0377] Methods far making bispecific antibodies are known in the art (e.g., c). Traditionally, the recombinant production of bispecific antibodies was based on the coexpression of two immunoglobulin heavy chain-light chain pairs, with the two heavy chains having different specificities (Millstein and Cuello, Hybrid hybridomas and their use in immunohistochemistry, Nature 1983, 305, 537-539).

[0378] Dosage Forms and Regimens

[0379] Administration of the compounds of the invention may be affected by any method that enables delivery of the compounds to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical, and rectal administration.

[0380] Those skilled in the art will be able to determine the appropriate amount, dose or dosage of each compound, as used in the combination ofthe present invention, to administerto a patient, taking into account variety of factors, including, though not limited to, the degree of advancement of the disease, age, weight, general health, gender, diet, the compound administered, the time and route of administration, the nature and advancement of cancer, requiring treatment, and other medications the individual is taking.

[0381] In some embodiments, the methods of administration of the agents and combinations herein may include oral, intravenous, intramuscular subcutaneous, topical, transdermal, intraperitoneal, intraorbital, by implantation, by inhalation, intrathecal, intraventricular, or intranasal administration.

[0382] Dosage regimens may be adjusted to provide the optimum desired response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are 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 art of compounding such an active compound for the treatment of sensitivity in individuals.

[0383] Thus, the skilled artisan would appreciate, based upon the disclosure provided herein, that the dose and dosing regimen is adjusted in accordance with methods well-known in the therapeutic arts. That is, the maximum tolerable dose can be readily established, and the effective amount providing a detectable therapeutic benefit to a patient may also be determined, as can the temporal requirements for administering each agent to provide a detectable therapeutic benefit to the patient. Accordingly, while certain dose and administration regimens are exemplified herein, these examples in no way limit the dose and administration regimen that may be provided to a patient in practicing the present invention.

[0384] For combination therapies as described herein, the agents may be administered at their approved dosages. Treatment is continued as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in certain embodiments, the combination therapies of the present invention may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the various monotherapies, it will be appreciated by the skilled practitioner that when the agents of the invention are used as part of a combination therapy, a lower dosage of the agent may be desirablethan when the agent alone is administered to a subject, a synergistic therapeutic effect may be achieved through the use of combination therapy which, in turn, permits use of a lower dose of the agent to achieve the desired therapeutic effect.

[0385] In one embodiment, the dosages may be lower and may also be applied less frequently, which may diminish the incidence or severity of side-effects. This is in accordance with the desires and requirements of the subjects to be treated.

[0386] It is one objective of this invention to provide a pharmaceutical composition comprising an amount, which may be jointly therapeutically effective at treating cancer. In this composition, two or more compounds may be administered together, one after the other or separately in one combined unit dosage form or in two separate unit dosage forms.

[0387] The unit dosage form may also be a fixed combination, it is to be noted that dosage values may vary with the type and severity of the condition to be alleviated and may include single or multiple doses, it is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. For example, doses may be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects such as toxic effects and / or laboratory values. Thus, the present invention encompasses intra-patient dose-escalation as determined by the skilled artisan. Determining appropriate dosages and regimens for administration of the chemotherapeutic agent are well-known in the relevant art and would be understood to be encompassed by the skilled artisan once provided the teachings disclosed herein.

[0388] The amount of the agent of the invention administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician.

[0389] An effective amount of the CDK inhibitor(s) and PD-1 axis binding antagonist, may be administered for prevention or treatment of disease. The appropriate dosage of the CDK inhibitor(s), PD-1 axis binding antagonist, may be determined based on the type of disease to be treated, the type of the CDK inhibitor, PD-1 axis binding antagonist, the severity and course of the disease, the clinical condition of the subject, the subject's clinical history and response to the treatment, and the discretion of the attending physician. In some embodiments, combination treatment with CDK inhibitor(s), and PD-1 axis binding antagonist (e.g., anti- PD-1 antibody or anti-PD-L1 antibody) are synergistic, whereby an efficacious dose of the CDK inhibitor (s) and PD-1 axis binding antagonist, in the combination is reduced relative to efficacious dose of the each of the CDK inhibitor (s), PD-1 axis binding antagonist, as a single agent.

[0390] Dosage units for a PD-1 axis binding antagonist (e.g., pembrolizumab, nivolumab, avelumab) may be expressed as a flat dose, i.e., 100 mg, 200 mg, 300 mg, or as a patient-specific dose, i.e., mg / kg (mg therapeutic agent / kg of body weight) or mg / m2(quantity in milligrams per square meter of body surface area).

[0391] As a general proposition, the therapeutically effective amount of the antibody administered to human will be in the range of about 0.01 to about 50 mg / kg of patient body weight whether by one or more administrations. In some embodiments, the antibody used is about 0.01 to about 45 mg / kg, about 0.01 to about 40 mg / kg, about 0.01 to about 35 mg / kg, about 0.01 to about 30 mg / kg, about 0.01 to about 25 mg / kg, about 0.01 to about 20 mg / kg, about 0.01 to about 15 mg / kg, about 0.01 to about 10 mg / kg, about 0.01 to about 5 mg / kg, or about 0.01 to about 1 mg / kg administered daily, for example. In some embodiments, the antibody is administered at 15 mg / kg. However, other dosage regimens may be useful. For example, in some embodiments, an anti-PD-L1 antibody described herein is administered to a human at a dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg or about 1400 mg on day 1 of 21 -day cycles. The dose may be administered as a single dose or as multiple doses (e.g., 2 or 3 doses), such as infusions. The dose of the antibody administered in a combination treatment may be reduced as compared to a single treatment. The progress of this therapy is easily monitored by conventional techniques.

[0392] In some embodiments that employ an antibody, antibody fragment or fusion soluble receptor as the PD-1 axis binding antagonist in the combination therapy, may comprise administering the antibody at a dose of about 0.5, 1, 2, 3, 5 or 10 mg / kg at intervals of about 7 days (± 2 days) or 14 days (± 2 days) or about 21 days (± 2 days) or about 30 days (± 2 days) throughout the course of treatment. Alternately, in some embodiments that employ an antibody, antibody fragment or fusion soluble receptor as the PD-1 axis binding antagonist in the combination therapy, the dosing regimen will comprise administering the antibody a dose of from about 0.005 mg / kg to about 10 mg / kg, with intrapatient dose escalation. In other escalating dose embodiments, the interval between doses will be progressively shortened, e.g., about 30 days (± 2 days) between the first and second dose, about 14 days (± 2 days) between the second and third doses. In certain embodiments, the dosing interval will be about 14 days (± 2 days), for doses subsequentto the second dose. In certain embodiments, the dosing interval will be about 7 days (± 2 days), for doses subsequent to the second dose.

[0393] In certain embodiments, a subject will be administered an intravenous (IV) infusion of a medicament comprising any of the PD-1 axis binding antagonists described herein.

[0394] In one embodiment of the invention, the PD-1 axis binding antagonist in the combination therapy is nivolumab, pembrolizumab or avelumab, which is administered intravenously or in a liquid dosage form at a dose selected from the group consisting of any one of: 1 mg / kg Q2W, 2 mg / kg Q2W, 3 mg / kg Q2W, 5 mg / kg Q2W, 10 mg Q2W, 1 mg / kg Q3W, 2 mg / kg Q3W, 3 mg / kg Q3W, 5 mg / kg Q3W, and 10 mg Q3W.

[0395] In some embodiments, pembrolizumab is administered at a dose of 2 mg / kg (up to 200 mg) every 3 weeks. In some embodiments, avelumab is administered at a dose of 10 mg / kg as an intravenous infusion over 60 minutes every 2 weeks. In some embodiments, the optimal dose for a PD-1 axis binding antagonist in combination with a CDK inhibitor may be identified by dose escalation of one or both of these agents.

[0396] The CDK4 inhibitor may be administered orally (PO), either once daily (QD) or twice daily (BID), with or without food on a continuous or intermittent schedule starting on Cycle 1 Day 1, A PD-1 axis binding antagonist such as avelumab may be administered as a 30-minute to 1 -hr intravenous (IV) infusion every 2 weeks (Q2W), every 3 weeks (Q3W) or in case of dose reduction, every 4 weeks (Q4W), starting on Cycle 1 Day 1. On the day of CDK inhibitor(s) administration, the CDK inhibitor may be given prior to or after administration of the PD-1 axis binding antagonist. In another embodiment, an CDK inhibitor(s) can be administered at 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg on a BID or QD schedule, which may be administered continuously or on an intermittent dosing schedule, such as 3 weeks on:1 week off (3:1) or 2 weeks on:1 week off (2:1) schedule, and the PD-1 axis binding antagonist is administered at a starting dose of 2 mg / kg, or 5 mg / kg or 10 mg / kg, at a dosing interval of Q2W, Q3Wor alternately Q4W.

[0397] In some embodiments, a treatment cycle begins with the first day of combination treatment and last for 3 weeks. In such embodiments, the combination therapy is preferably administered for at least 18 weeks (6 cycles of treatment), more preferably at least 24 weeks (8 cycles of treatment), and even more preferably at least 2 weeks after the patient achieves a CR.

[0398] Dosage units for a CDK inhibitor (e.g., atirmociclib, PF-Q7104091 or palbociclib) may be expressed as a flat dose, doses for each CDK inhibitor is independently of one another, i.e., 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg, etc. or as a subject-specific dose, i.e., mg / kg (mg therapeutic agent / kg of body weight) or mg / m2(quantity in milligrams per square meter of body surface area).

[0399] Some embodiments may comprise administering the CDK inhibitor(s), doses for each CDK inhibitor is independently of one another, in a dose of about: 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 150 mg, 150 mg, 175 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg, mg, wherein the amounts can be administered once a day (q.d.), twice a day (b.i.d), three times a day (t.i.d.), four times a day (q.i.d.), or on some other dosing schedule.

[0400] All the doses refer to the free base equivalent.

[0401] Repetition of the administration or dosing regimens, or adjustment of the administration or dosing regimen may be conducted as necessary to achieve the desired treatment. A “continuous dosing schedule,” as used herein, is an administration or dosing regimen without dose interruptions, e.g., without days off treatment. Repetition of 21 - or 28-day treatment cycles without dose interruptions between the treatment cycles is an example of a continuous dosing schedule. In an embodiment, the compounds of the combination of the present invention can be administered in a continuous dosing schedule.

[0402] This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.

[0403] The compounds of the combination of the present invention can be administered intermittently, concurrently or sequentially. In an embodiment, the compounds of the combination of the present invention can be administered in a concurrent dosing regimen,

[0404] i. Kits

[0405] The invention also provides a kit comprising: a. (i) a pharmaceutical composition comprising a CDK4 inhibitor of Formula (I) and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-1 binding antagonist and a pharmaceutically acceptable carrier;

[0406] b. (I) a pharmaceutical composition comprising a CDK4 inhibitor of Formula (I) and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-1 binding antagonist and a pharmaceutically acceptable carrier; (iii) a pharmaceutical composition comprising PF-07104091 and a pharmaceutically acceptable carrier;

[0407] c. (I) a pharmaceutical composition comprising atirmociclib and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-1 binding antagonist and a pharmaceutically acceptable carrier; or

[0408] d. (i) a pharmaceutical composition comprising atirmociclib and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-1 binding antagonist and a pharmaceutically acceptable carrier; (iii) a pharmaceutical composition comprising PF- 07104091 and a pharmaceutically acceptable carrier;

[0409] e. (i) a pharmaceutical composition comprising a CDK4 inhibitor of Formula (I) and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-L1 binding antagonist and a pharmaceutically acceptable carrier;

[0410] f. (i) a pharmaceutical composition comprising a CDK4 inhibitor of Formula (I) and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-L1 binding antagonist and a pharmaceutically acceptable carrier; (iii) a pharmaceutical composition comprising PF-07104091 and a pharmaceutically acceptable carrier;

[0411] g. (I) a pharmaceutical composition comprising atirmociclib and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-L1 binding antagonist and a pharmaceutically acceptable carrier; or

[0412] h. (i) a pharmaceutical composition comprising atirmociclib and a pharmaceutically acceptable carrier; (ii) a pharmaceutical composition comprising a PD-L1 binding antagonist and a pharmaceutically acceptable carrier; (iii) a pharmaceutical composition comprising PF-07104091 and a pharmaceutically acceptable carrier.

[0413] The PD-1 axis binding antagonist (i.e., PD-1 binding antagonist or PD-L1 binding antagonist) are described herein.

[0414] in some embodiments, the PD-1 axis binding antagonist is provided in a container. Suitable containers include, for example, bottles, vials, bags and syringes. The container may be formed from a variety of materials such as glass, plastic (such as polyvinyl chloride or polyolefin), or metal alloy (such as stainless steel or hastelloy). in some embodiments, the container holds the formulation and the label on, or associated with, the container may indicate directions for use. The kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use. in some embodiments, the kit further includes one or more of another agent (e.g., a chemotherapeutic agent, and anti-neoplastic agent). Suitable containers for the one or more agent(s) include, for example, bottles, vials, bags and syringes.

[0415] The specification is sufficient to enable one skilled in the art to practice the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0416] EXAMPLES

[0417] The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention. It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims.

[0418] Example 1: The CDK4 Selective inhibitor atirmociclib and the CDK2 Selective Inhibitor tegtociclib Synergistically Upreguiate PD-L1 Protein at the Cell Surface of Tumor Cells.

[0419] Overview

[0420] CDK4 / 6 inhibitors can augment anti-tumor immunity through a variety of cancer- and immune-cell dependent mechanisms, including improved antigen presentation and pro- inflammatory cytokine signaling by tumor cells.

[0421] Atirmociclib (“At”) was evaluated for its potential to upregulate expression of genes involved in antigen presentation and pro-inflammatory cytokine signaling in a selected panel of tumor cell lines. In addition, atirmociclib was evaluated in combination with a CDK2 inhibitor PF- 07104091 for synergistic efficacy in upregulating these genes, including PDL1.

[0422] Materials and Methods

[0423] HR+ HER2- breast cancer cell lines MCF7, MCF7 Palbo Resistance (MCF7PR), T47D, and HCC1428 (FIG. 1) or lung adenocarcinoma cell lines HCC78, H2087, H1573, H1792, HCC2108 (FIG. 2) were seeded in 10-cm dish, and treated with vehicle (Veh), 30 nM palbociclib, 1 pM atirmociclib (At), 1 pM CDK2 inhibitor PF-07104091 (CDK2I), or the combination (At +CDK2I). Celis were treated for 8 days and the culture media was replenished with new added drug every 2-3 days. The cell confluence was 80% in all the groups at the end point. The surface expression of MHCI and PD-L1 was detected by flow cytometric analysis. The PD-L1 antibody was CD274 (PD-L1, B7-H1) Monoclonal Antibody (MIH1 ), APC, eBioscience, Invitrogen, and the MHCI antibody was HLA-ABC Monoclonal Antibody (W6 / 32), FITC, eBioscience, Invitrogen. For gene expression, total RNA was extracted using the Quick-RNA Miniprep Plus Kit (Zymo Research) according to the manufacturer’s instructions. The RNA was treated with DNase I. cDNA was synthesized with the iScript cDNA synthesis kit (Bio-Rad). qPCR was performed using the SYBR Green master mix (Bio-Rad). Gene expression level was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. Fold change represents the average fold change compared to the control condition (n=2). The primers used in the study are listed in Table 4 below.

[0424] Table 4

[0425] Gene Forward primer Reverse primer

[0426] HLA-A 5'-ACCCTCGTCCTGCTACTCTC-3' 5-CTGTCTCCTCGTCCCAATACT-3' HLA-B 5'-CAGTTCGTGAGGTTCGACAG-3' 5’-CAGCCGTACATGCTCTGGA-3’ HLA-C 5'-GGACAAGAGCAGAGATACACG-3' 5'-CAAGGACAGCTAGGACAACC-3' B2M 5’-GAGGCTATCCAGCGTACTCCA~3' 5'-CGGCAGGCATACTCATCTTTT-3’ TAP1 5’-TTTGAGTACCTGGACCGCAC-3’ 5-AATGTCAGCCCCTGTAGCAC-3’ TAP2 5’-TCAAGGGGCTGACGTTTACC-3’ 5’-AGAACCGGAGAACAGCACAG-3’ TAPBP 5’-GACAGCGTAGGCCTTTTCCT-3’ 5’-AGGTTCTGCCTACCACACCA-3’ (variant 2)

[0427] TAPBP 5’-AGTGTACACGTGGTACTGACTG- 5’-GATGTCCCTTACCCAGGTGC-3’ (variant 3) 3’

[0428] PD-L1 5'-TGGCATTTGCTGAACGCATTT“3' 5'-TGCAGCCAGGTCTAATTGTTTT-3' STAT1 5’-AACCTGACTTCCATGCGGTT-3’ 5-GAGCAGGTTGTCTGTGGTCT-3’ STAT2 5’-TCGAAACACCTGTGGAGAGC-3’ 5’-GTCTTCCCTTTGGCCTGGAT-3’ IRF9 5’-TCTGCAGAGACTTGGTCAGG-3’ 5’-CTGCTCCAGCAAGTATCGGG-3’ NLRC5 5’-GCTGTGGTCTCTCAGCAAAC-3’ 5’-GCAGACTTCTAACAGGGAGAG-3’ DNMT1 5’-GAGGAGGGCTACCTGGCTAA-3’ 5’-GCTTAGCCTCTCCATCGGAC-3’ GAPDH 5'-GGAGCGAGATCCCTCCAAAAT-3' 5'-GGCTGTTGTCATACTTCTCATGG-3'

[0429]

[0430] Results

[0431] Both atirmociciib alone and the combination of atirmociclib and tegtociclib (CDK2i) increased antigen processing and presentation, as well as interferon pathway gene expression in tumor ceils. In contrast, palbociciib at clinically relevant concentration (CRC) had minimal effects. The surface expression of PDL1 and MHCi increased following treatment with atirmociclib and combination of atirmociclib and CDK2I. The surface expression of PDL1 is enriched in senescent tumor cells induced by atirmociclib and the combination of atirmociciib and CDK2I treatment. Example 2: The CDK4 Selective Inhibitor atirmociclib Synergizes with PD-1 Based Immune Checkpoint Blockade in MC38R and CT26 Syngeneic Mouse Tumor Models

[0432] Overview

[0433] Atirmociclib was evaluated in combination with a PD-1 antibody in the MC38R and CT26 syngeneic mouse tumor models to assess efficacy of in vivo tumor growth inhibition. Moreover, atirmociclib and palbociclib were compared at their clinically relevant doses for tumor growth inhibition combination efficacy with the PD-1 antibody.

[0434] Materials and Methods

[0435] MC38R tumor-bearing mice were established by implanting 0.5 x 106tumor cells in 0.2 ml of PBS into the dorsal region of female C57BL / 6 J mice (Jackson Lab). For efficacy testing, mice were randomly assigned to groups (n =12 per group) once tumor volumes reached between 40 mm3and 133 mm3. They were then treated with (1) vehicle (0.5% (w / v) methylcellulose (A4M) in deionized water), PO, BID; (2) PD1 antibody (PF-06937004) at 10 mg / kg Q3D x4, IP; (3) atirmociclib at 60 mg / kg BID, PO; (4) atirmociclib at 60 mg / kg BID, PO plus PD1 antibody (PF- 06937004) at 10 mg / kg Q3D x4, IP; (5) palbociclib at 10 mg / kg BID, PO; (6) palbociclib at 10 mg / kg BID, PO plus PD1 antibody (PF-06937004) at 10 mg / kg Q3D x4, IP. All groups received treatment continuously until Day 28 (inclusive).

[0436] CT26 tumor-bearing mice were established by implanting 0.2 x 105tumor cells in 0.2 ml of PBS into the dorsal region of female Balb / C J mice (Jackson Lab). For efficacy testing, mice were randomly assigned to groups (n = 12 per group) once tumor volumes reached between 50 mm3and 123 mm3. Treatment arms were same as for MC38R animals described above but animals received treatment continuously until Day 15 (inclusive).

[0437] The treatment groups and dose regimen information are summarized in Table 5.

[0438] Table 5

[0439] Group Drug n Route Regimen 1 vehicle 12 PO BID continuously 2 PF-06937004 (clone F2) 10 mg / kg 12 IP Q3D; 4 doses 3 Atirmociclib 60 mg / kg 12 PO BID continuously Atirmociclib 60 mg / kg + PO (At) BID continuously (At) 4 12

[0440] PF-06937004 (clone F2) 10 mg / kg IP (aPD1) Q3D; 4 doses (aPD1) 5 Palbociclib 10 mg / kg 12 PO BID continuously Palbociclib 10 mg / kg + PO (Pa) BID continuously (Pa) 6 12

[0441] PF-06937004 (clone F2) 10 mg / kg IP (aPD1) Q3D; 4 doses (aPD1)

[0442]

[0443] BID = twice daily; IP = intraperitoneal injection; PO = oral dosing; Q3D = 1 dose every 3 days; n = number of animals per group

[0444] Tumor volumes were measured three times a week. Tumor volumes were calculated based on two-dimensional caliper measurement with cubic millimeter volume calculated using the formula (length x width2) x 0.5. Mice were sacrificed when the tumor volumes reached 2000 mm3, which was the survival endpoint for this study. Tumor volumes were plotted using GraphPad Prism 7 software. Statistical significance determined using the Holm-Sidak method, with alpha = 0.05.

[0445] Results

[0446] On Day 16 (MC38) and day 13 (CT26) post-treatment initiation, the tumor growth results show that monotherapy with CDK4 inhibitor (atirmociclib) inhibited tumor growth in both MC38R and CT26 models (p < 0.05), more effectively than either the anti-PD-1 antibody (PF-06937004 clone F2) monotherapy or CDK4 / 6 inhibitor (palbociclib) monotherapy.

[0447] However, the combination of the CDK4 inhibitor (atirmociclib) with the anti-PD-1 antibody (PF-06937004 clone F2) demonstrated a strong combinatorial effect, resulting in the most significant tumor growth inhibition (p < 0.05). These data are summarized as mean tumor volume in FIGS. 5-8, with absolute values presented in Table 6.

[0448] Table 6

[0449] % TGI % TGI

[0450] Group | Drug

[0451] MC38R CT2620

[0452] 1 | vehicle

[0453] 2 | PF-06937004 (clone F2) 10 mg / kg 38 17

[0454] 3 | Atirmociclib 60 mg / kg 64 42

[0455] | Atirmociclib 60 mq / kq + 25

[0456] 4 80 63

[0457] PF-06937004 (clone F2) 10 mg / kg

[0458] 5 | Palbociclib 10 mg / kg 31 25

[0459] | Palbociclib 10 mg / kg +

[0460] 44 35

[0461] PF-06937004 (clone F2) 10 mg / kg

[0462]

[0463] _ 3£L_

[0464] Conclusion

[0465] Synergistic tumor growth inhibition efficacy was seen when atirmociclib was combined with immune checkpoint blockade antibodies in MC38R and CT26 syngeneic mouse tumor models. Moreover, in both models, when atirmociciib and palbociclib were used at their clinically relevant doses, the combination of atirmociciib and Immune checkpoint blockade antibodies led to greater tumor growth inhibition than the combination of palbociclib and immune checkpoint blockade antibodies.

[0466] All references cited herein, including patent applications, patent publications, and UniProtKB / Swiss-Prot Accession numbers cited in the specification are herein incorporated by reference in their entirety. Although the foregoing invention has been described in some detail by way of illustration and example, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0467] The foregoing description and Examples detail certain specific embodiments of the invention and describes the best mode contemplated by the inventors. It will be appreciated, however, that no matter how detailed the foregoing may appear in text, the invention may be practiced in many ways and the invention should be construed in accordance with the appended claims and any equivalents thereof.

Claims

CLAIMSWhat is claimed:

1. A method for treating cancer in a subject in need thereof comprising administering to a subject(a) an effective amount of a CDK4 inhibitor having a Formula (I):wherein:R1is H, F or Cl;R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R5;R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R6; R4is H or F; andeach R5and R6is independently OH, F or C1-C2 alkoxy; and(b) an effective amount of a PD-1 axis binding antagonist; and(c) optionally an effective amount of a CDK2 inhibitor.

2. The method of claim 1, wherein the CDK4 inhibitor of Formula (I) is atirmociclib.

3. The method of claim 1, wherein the CDK2 inhibitor is tegtociclib.

4. The method of any one of claims 1 to 3, wherein the PD-1 axis binding antagonist comprises a PD-1 binding antagonist, ora PD-L1 binding antagonist.

5. The method of any one of claims 1 to 4, wherein the PD-1 axis binding antagonist comprises a PD-1 binding antagonist.

6. The method of claim 5, wherein the PD-1 binding antagonist is an anti-PD-1 antibody.

7. The method of claim 6, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, pidilizumab, cemiplimab, tislelizumab, spartalizumab, RN888, mAb15, MEDI-0680, BGB-108, AGEN-2034, or a combination thereof.

8. The method of claim 3, wherein the PD-1 axis binding antagonist comprises a PD-L1 binding antagonist.

9. The method of any one of claims 1 to 4, wherein the PD-L1 binding antagonist is an anti- PD-L1 antibody.

10. The method of claim 9, wherein the anti-PD-L1 antibody is BMS-936559, AMP-714, atezoiizumab, durvalumab, avelumab, toripaiimab, retifanlimab, dostariimab, IBI318, IMM251Q, or an antibody comprising a VH region produced by the expression vector with ATCC Accession No. PTA-121183 and having the VL region produced by the expression vector with ATCC Accession No. PTA-121182, or a combination thereof.

11. The method of any one of claims 1 to 10, wherein the subject is a human.

12. The method of any one of claims 1 to 11, wherein the cancer is selected from the group consisting of brain cancer, head / neck cancer, prostate cancer, ovarian cancer, bladder cancer, lung cancer, breast cancer, bone cancer, colorectal cancer, kidney cancer, liver cancer, stomach cancer, pancreatic cancer, esophageal cancer, cervical cancer, sarcoma, skin cancer, mesothelioma, malignant rhabdoid tumors, neuroblastoma, diffuse intrinsic pontine glioma (DIPG), and carcinoma.

13. The method of any one of claims 1 to 12, wherein the cancer is breast cancer.

14. The method of any one of claims 1 to 13, wherein the cancer is breast cancer selected from any one or more of: hormone receptor positive (HR+), hormone receptor negative (HR-), human epidermal growth factor receptor 2 negative (HER2-), human epidermal growth factor receptor 2 positive (HER2+), HR+ / HER2-, ER- / HR+, ER+ / HER2- and triple negative breast cancer (TNBC).

15. The method of any one of claims 1 to 12, wherein the cancer is NSCLC, prostate, colorectal cancer, liposarcoma, or tumors characterized by amplification or overexpression of CDK4 and / or CCND1.

16. A combination comprising:(iv) a CDK4 inhibitor having a Formula (I):wherein:R1is H, F or Cl;R2is C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R5;R3is H or C1-C4 alkyl, where said C1-C4 alkyl is optionally substituted by R5;R4is H or F; andeach R5and R6is independently OH, F or C1-C2alkoxy;(v) a PD-1 axis binding antagonist; and(vi) optionally tegtociclib;for use in the treatment of cancer in a subject.

17. The combination of claim 16, wherein the PD-1 binding antagonist is an anti-PD-1 antibody.

18. A combination comprising:a. (I) atirmociclib; and (ii) a PD-L1 axis binding antagonist; orb. (I) atirmociclib; (ii) PD-1 axis binding antagonist; and (ill) tegtociclib; for use in the treatment of cancer in a subject.

19. The combination of claim 18, wherein the PD-L1 binding antagonist is an anti-PD-L1 antibody.

Images