RU2796903C2 - Combination of hdac inhibitor and anti-pd-1 antibody for treatment of cancer - Google Patents

Abstract

FIELD: medicine; oncology.

SUBSTANCE: patient with melanoma or non-small cell lung cancer is administered with a combination of therapeutic agents: entinostat and pembrolizumab. In this case, the patient has previously received a PD-1-blocking antibody or a PD-L1-blocking antibody and has progression in previous therapy with the PD-1-blocking antibody or the PD-L1-blocking antibody. Entinostat is administered orally at a dose of 3 mg or 5 mg once a week. Pembrolizumab is administered intravenously at a dose of 200 mg every three weeks, while the duration of a treatment cycle is 21 days.

EFFECT: method allows for an increase in the efficiency of treatment of oncological diseases due to combination and dosage regimen of drugs.

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Description

CROSS REFERENCES

This application claims priority based on U.S. Provisional Application No. 62/136303, filed March 20, 2015, and U.S. Provisional Application 62/137601, filed March 24, 2015, which are incorporated herein by reference in their entirety.

SUMMARY OF THE INVENTION

In one embodiment, a method of treating cancer is provided, wherein the method comprises administering to a patient a combination containing an entinostat and an anti-PD-1 or anti-PD-L1 antibody. In some embodiments, the method includes administering to a patient a combination containing an entinostat and an anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody is pembrolizumab. In some embodiments, the cancer is characterized by overexpression of PD-L1. In some embodiments, the cancer is characterized by low expression of PD-L1. In some embodiments, the cancer is lung cancer or melanoma. In some embodiments, the patient has received at least one prior therapy. In some embodiments, the prior therapy was with an anti-PD-1 antibody. In some embodiments, the cancer is lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is selected from adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. In some embodiments, the cancer is melanoma. In some embodiments, the melanoma is unresectable or metastatic melanoma. In some embodiments, the patient has progressed on prior anti-CTLA4 antibody therapy. In some embodiments, the patient has progressed on prior therapy with a BRAF inhibitor. In some embodiments, the entinostat and the anti-PD-1 or anti-PD-L1 antibody are administered sequentially in any order or simultaneously. In some embodiments, the anti-PD-1 or anti-PD-L1 antibody is administered sequentially in any order or simultaneously during a 21 day treatment cycle. In some embodiments, the entinostat and the anti-PD-1 antibody are administered sequentially in any order or simultaneously. In some embodiments, the entinostat and the anti-PD-1 antibody are administered sequentially in any order or simultaneously during a 21 day treatment cycle. In some embodiments, the anti-PD-1 antibody is administered on day 1 of the treatment cycle. In some embodiments, the anti-PD-1 antibody is administered at a dose of 2 mg/kg. In some embodiments, the anti-PD-1 antibody is administered at a fixed dose of 200 mg. In some embodiments, the entinostat is administered at a dose of 3 mg/kg. In some embodiments, the entinostat is administered at a dose of 5 mg/kg. In some embodiments, the entinostat is administered at a dose of 10 mg/kg. In some embodiments, the entinostat is administered weekly. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered biweekly.

In a further embodiment of the invention, a method of treating cancer is provided, wherein the method comprises administering to a patient a combination comprising essentially entinostat and pembrolizumab. In some embodiments, the entinostat is administered as a solid dosage form and pembrolizumab is administered as an intravenous infusion. In some embodiments, the cancer is lung cancer or melanoma.

In another embodiment, the present invention provides a kit for the treatment of advanced non-small cell lung cancer and melanoma, comprising a combination of an entinostat and an anti-PD-1 or anti-PD-L1 antibody.

In another embodiment, the present invention provides a method for selecting a patient for combination therapy comprising administering an entinostat and an anti-PD-1 or anti-PD-L1 antibody, the method comprising measuring PD-L1 expression in a tumor tissue sample obtained from the patient.

In some embodiments, the method further comprises administering to the patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody if the tumor proportion score (TPS) for PD-L1 expression is greater than or equal to 50%. In some embodiments, the method further comprises administering to the patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody if the tumor proportion score (TPS) for PD-L1 expression is greater than or equal to 1%. In some embodiments, the method further comprises administering to the patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody if the tumor proportion score (TPS) for PD-L1 expression is between 1% and 50%. In some embodiments, the method further comprises administering to the patient a combination therapy comprising administering to the patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody if the tumor proportion score (TPS) for PD-L1 expression is greater than or equal to 49%. In some embodiments, the tumor tissue sample is obtained from squamous or non-squamous non-small cell lung cancer. In some embodiments, the tumor tissue sample is obtained from a melanoma.

INCLUSION BY LINK

All publications, patents, and patent applications described in this specification are incorporated by reference to the same extent as if each individual publication, patent, or patent application is specifically or individually incorporated by reference.

DETAILED DESCRIPTION

New strategies are needed for the treatment of cancer, in particular unresectable or metastatic melanoma and non-small cell lung cancer, which are not progressing satisfactorily with treatments currently known. Surprisingly, combination therapy using epigenetic modulation with histone deacetylase (HDAC) inhibitors such as entinostat and immune response checkpoint inhibitors such as anti-PD-1 or anti-PD-L1 antibodies has been found to provide an effective cancer treatment strategy, in particularly melanoma and non-small cell lung cancer.

Provided herein are methods for treating cancer based on the administration of an HDAC inhibitor and an anti-PD-1 antibody or an anti-PD-L1 antibody. In some embodiments, the HDAC inhibitor is an entinostat. In some embodiments, the anti-PD-1 antibody is pembrolizumab. The methods further include treatment wherein the combinations are supplemented with one or more therapeutic agents or therapy.

To facilitate understanding of the description provided here, a number of terms are defined below.

As used herein, "abnormal cell growth" refers to cell growth that is independent of normal regulatory mechanisms (eg, loss of contact inhibition), including abnormal growth of normal cells and growth of pathological cells.

"Neoplasia", as described herein, is an abnormal, unregulated, or disorganized proliferation of cells that differ from normal cells by autonomous growth and somatic mutations. As tumor cells grow and divide, they pass on their genetic mutations and proliferative characteristics to their descendants. A neoplasia or tumor is a collection of tumor cells. In some embodiments, the neoplasms may be benign or malignant.

"Metastasis", as used herein, refers to the dissemination of tumor cells through the lymphatic or blood vessels. Metastasis also refers to the migration of tumor cells by spreading directly through the serous cavities or subarachnoid or other spaces. Through the process of metastasis, tumor cells migrating to other areas of the body create neoplasms in areas far from their original site of origin.

As discussed herein, "angiogenesis" predominates in tumor formation and metastasis. Angiogenic factors have been found to be associated with several solid tumors such as rhabdomyosarcomas, retinoblastoma, Ewing's sarcoma, neuroblastoma and osteosarcoma. A tumor cannot spread without a blood supply to provide nutrients and remove cellular waste. Tumors in which angiogenesis is important include solid tumors such as renal cell carcinoma, hepatic cell carcinoma, and benign tumors such as acoustic neuroma and neurofibroma. Angiogenesis is associated with tumors of the hematopoietic system, such as leukemia. It is believed that angiogenesis plays a role in the pathology in the bone marrow that leads to the development of leukemia. Prevention of angiogenesis can stop the growth of cancerous tumors and the resulting harm to the patient due to the presence of the tumor.

The term "patient" refers to an animal, including, without limitation, a primate (eg, human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject" and "patient" are used interchangeably in relation to, for example, a mammalian patient, such as a human patient.

The terms "treat" "treatment" and "therapy" include the alleviation or elimination of a disorder, disease, or condition; or one or more symptoms associated with the disorder, disease, or condition; or alleviate or eliminate the cause(s) of the disorder, disease, or condition as such.

The term "therapeutically effective amount" refers to an amount of a compound that, when administered, is sufficient to prevent the development, or alleviate to some extent, one or more symptoms of the disorder, disease, or condition being treated. The term "therapeutically effective amount" also refers to the amount of a compound that is sufficient to produce a biological or medical response of a cell, tissue, system, animal or human, as selected by the researcher, veterinarian, physician or clinician.

The term "pharmaceutically acceptable carrier,""pharmaceutically acceptable excipient,""physiologically acceptable carrier," or "physiologically acceptable excipient" refers to a pharmaceutically acceptable material, composition, or carrier such as a liquid or solid excipient, diluent, excipient, solvent, or encapsulating material. Each component must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical composition. It must also be suitable for use in contact with a tissue or organ in humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problems or complications, according to a reasonable benefit/risk ratio. See, Remington: The Science and Practice of Pharmacy , 21st Edition; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients , 5th Edition; Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association: 2005; and Handbook of Pharmaceutical Additives , 3rd Edition; Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation , Gibson Ed., CRC Press LLC: Boca Raton, FL, 2004).

The term "pharmaceutical composition" refers to a mixture of a compound described herein with other chemical components such as diluents and carriers. The pharmaceutical composition facilitates the administration of the compound into the body. Multiple methods for administering a compound exist in the art, including, without limitation, oral, injectable, aerosol, parenteral, and topical administration. Pharmaceutical compositions can be prepared by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

Cancer, tumors, tumor-related disorders, and neoplastic pathological conditions are serious and often life-threatening conditions. Such diseases and disorders, which are characterized by rapidly proliferating cell growth, continue to be the subject of research aimed at identifying therapeutic agents that are effective in their treatment. Such agents prolong patient survival, inhibit the growth of rapidly proliferating cells associated with neoplasia, or cause regression of neoplasia.

HDAC inhibitors represent a promising class of therapeutic agents that drive differentiation and apoptosis in hematological and solid neoplasms through chromatin remodeling and regulation of gene expression. Several HDAC inhibitors have been identified, including benzamides (entinostat), short chain fatty acids (i.e. sodium phenylbutyrate); hydroxamic acids (ie siberoylanilide hydroxamic acid and trichostatin A); cyclic tetrapeptides containing a 2-amino-8-oxo-9, 10-epoxy-decanoyl moiety (i.e. trapoxin A) and cyclic peptides without a 2-amino-8-oxo-9,10-epoxy-decanoyl moiety (t .e. FK228). Entinostat is a benzamide HDAC inhibitor undergoing clinical trials in many types of solid tumors and hematologic cancers. Entinostat is rapidly absorbed and has a half-life of about 100 hours and, importantly, changes in histone acetylation persist for several weeks after entinostat administration.

High expression of PD-L1 on tumor cells has been found to correlate with poor prognosis and survival in various types of solid tumors. Without wishing to be bound by any theory, it is contemplated that the PD-1/PD-L1 pathway plays a critical role in tumor evasion from the immune response and may be an attractive target for therapeutic intervention in several types of solid tumors.

Several PD-1 and PD-L1 antibodies are in clinical development. In general, they are reported to be well tolerated, for the most part, without reaching dose-limiting toxicity in phase I studies.

Histone deacetylase

HDACs are a family of at least eighteen enzymes grouped into three classes (class I, II and III). Class I HDACs include, without limitation, HADCs 1, 2, 3, and 8. Class I HDACs can be found in the nucleus and are considered to be involved in transcriptional control repressors. Class II HDAC includes, without limitation, HDACS 4, 5, 6, 7, and 9 and can be found in both the cytoplasm and the nucleus. Class III HDACs are considered to be NAD-dependent proteins and include, without limitation, members of the sirtuin family of proteins. Non-limiting examples of sirtuin proteins include SIRT1-7. As used herein, the term "selective HDAC" refers to an HDAC inhibitor that does not interact with all three classes of HDAC.

HDAC inhibitors

HDAC inhibitors can be broadly classified into pan-HDAC inhibitors and selective HDAC inhibitors. Although there are major structural differences between known HDAC inhibitors, they share common characteristics: a moiety that interacts with the active site of the enzyme and a side chain that is located within the channel leading to the active site. This can be seen in hydroxamates such as SAHA where the hydroxamate group is thought to interact with the active site. In the case of depsipeptides, intracellular disulfide bond reduction is believed to create a free thiol group (which interacts with the active site) attached to the 4-carbon alkenyl group. The difference between HDAC inhibitors is that they can interact with the rim of the HDAC channel, which is located at the opposite end of the channel from the active site. This interaction between the HDAC inhibitor and the channel rim is thought to account, at least in part, for some of the observed differences in HDAC selectivity between pan-HDAC inhibitors such as SAHA and selective HDAC inhibitors such as depsipeptides. A particularly preferred HDAC inhibitor is entinostat. Enthinostat has the chemical name N-(2-aminophenyl)-4-[N-(pyridin-3-yl)methoxycarbonylaminomethyl]benzamide and the chemical structure shown below.

Chemical structure of entinostat

Programmed cell death-1 (PD-1)

PD-1 is a cell surface receptor that is a member of the CD28 T cell regulator family, within the immunoglobulin receptor superfamily. The human PD-1 gene is located on chromosome 2q37, and the full-length PD-1 cDNA encodes a 288 amino acid residue protein with 60% homology to mouse PD-1. It is present on CD4CD8 (double negative) thymocytes during thymus development and is expressed upon activation in mature hematopoietic cells such as T and B cells, NKT cells, and monocytes after prolonged antigen exposure.

PD-L1 has been shown to be expressed on a number of murine and human tumors (and induced by IFN-gamma on most PD-L1 negative tumor cell lines) and has been shown to mediate immune surveillance escape (Iwai Y. et al., Proc. Natl. Acad. Sci. U.S.A. 99: 12293-12297 (2002) Strome S. E. et al., Cancer Res., 63: 6501-6505 (2003) In humans, PD-1 and/or PD-L1 expression has been detected in a number of biopsies primary cancers of the lung, liver, ovary, cervix, skin, colon, gliomas, bladder, breast, kidney, esophagus, stomach, oral squamous cell, urothelial cells, and pancreas, as well as head and neck tumors (Brown J. A. et al., J. Immunol. 170: 1257-1266 (2003), Dong H. et al., Nat. Med. 8: 793-800 (2002), Wintterle et al., Cancer Res. 63: 7462- 7467 (2003), Strome S. E. et al., Cancer Res., 63: 6501-6505 (2003), Thompson R. H. et al., Cancer Res. 66: 3381-5 (2006), Thompson et al., Clin. Cancer Res. 13: 1757-61 (2007); Nomi T. et al., Clin. Cancer Res. 13:2151-7. (2007)). Expression of PD ligand on tumor cells has been correlated with poor prognosis in cancer patients in a variety of tumor types (described in Okazaki and Honjo, Int. Immunol. 19: 813-824 (2007)).

Several studies have shown that the interaction of PD-1 with its ligands (PD-L1 and PD-L2) results in inhibition of lymphocyte proliferation in vitro and in vivo . Accordingly, without wishing to be bound by any theory, it is believed that the PD-L1 anti-PD-1 ligand suppresses effector antitumor T-cell activity and facilitates evasion of the immunological response.

Disruption of the PD-1/PD-L1 interaction has been shown to increase T cell proliferation and cytokine production and block cell cycle progression. In vitro studies of PD-1 blockade with a PD-1 specific antibody have shown an increase in cytotoxic T-cell responses to melanoma-specific antigens, including an increased frequency of IFN-γ-secreting antigen-specific cells.

Without being bound by any theory, it is believed that targeting PD-1 may act as an effective therapeutic strategy for cancer.

The main method for targeting PD-1 has been clinically accomplished through the development of engineered monoclonal antibodies that inhibit the function of either PD-1 or PD-L1.

The methods of the present description include the use of full-length antibodies to PD-1 (anti-PD-1 antibody) and its antigen-binding portion. Examples of binding fragments encompassed by the term "antigen-binding portion thereof" include (i) a Fab fragment, a monovalent fragment consisting of the Vi, V//, CL, and C domains; (ii) an F(ab')2 fragment, a divalent fragment comprising two Fab fragments linked by a disulfide bridge in a loop region; (iii) Fd fragment consisting of \1H and Cm domains; and (iv) an Fv fragment consisting of Vi and V// domains on one Ab arm.

The anti-PD-1 antibody and its antigen-binding portion bind to PD-1 with high specificity and affinity, block the binding of PD-L1 and/or PD-L2, and inhibit the immunosuppressive effect of the PD-1 signaling pathway. In some embodiments, the combination therapy comprises administering an entinostat and an anti-PD-1 antibody, or antigen-binding portion thereof, wherein the antibody, or antigen-binding portion thereof, is a chimeric, humanized, or human monoclonal antibody, or portion thereof. In certain embodiments, the anti-PD-1 antibody, or antigen-binding portion thereof, is a humanized antibody. In some embodiments, the anti-PD-1 antibody, or antigen-binding fragment thereof, is a human antibody. In some embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is a monoclonal antibody or antigen-binding portion thereof.

Without wishing to be bound by theory, it is contemplated that an anti-PD-L1 antibody prevents interaction between PD-1 and PD-L1, thus having similar effects in the PD-1 signaling pathway. Accordingly, in some embodiments, the combination therapy comprises the administration of an entinostat and an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody, or antigen-binding portion thereof, binds specifically to the programmed death ligand 1 (PD-L1) receptor and inhibits PD-L1 activity.

pembrolizumab

Pembrolizumab is a humanized anti-PD-1 monoclonal antibody consisting of a high affinity mouse anti-PD-1 derived variable region grafted onto a human IgG4 immunoglobulin molecule with an Fc engineered region for stabilization. Pembrolizumab has been approved by the FDA for the treatment of unresectable or metastatic melanoma and metastatic non-small cell lung cancer (NSCLC).

Lung cancer

Lung cancer is the leading cause of cancer death in women and men both in the United States and worldwide. Lung cancer has overtaken breast cancer as the leading cause of cancer death in women. In the United States, 158,040 people were estimated to have died of lung cancer in 2014, more than the number of deaths from colon and rectal, breast, and prostate cancers combined. Only about 2% of those diagnosed with lung cancer that has spread to other areas of the body live five years after diagnosis, although the survival rate for lung cancer diagnosed in the earliest stages is higher, with approximately 49% survival for five years or more.

Cancer develops when normal cells undergo a transformation that allows them to grow and multiply out of control. The cells form a mass or tumor that is different from the surrounding tissues from which it originates. Tumors are dangerous because they consume oxygen, nutrients and space from healthy cells, and because they invade and destroy or reduce the ability of normal cells to function.

Most lung tumors are malignant. This means that they invade and destroy healthy tissues around them and can spread throughout the body. Tumors can spread to nearby lymph nodes or through the bloodstream to other organs. This process is called metastasis. When lung cancer metastasizes, the tumor in the lung is called the primary tumor, and tumors elsewhere in the body are called secondary tumors or metastatic tumors.

Some tumors in the lung are metastatic from cancers elsewhere in the body. The lungs are a common site of metastasis. In this case, the cancer is not considered lung cancer. For example, if prostate cancer spreads through the bloodstream to the lungs, it is metastatic prostate cancer (secondary cancer) in the lung and is not called lung cancer.

Lung cancer includes a group of different types of tumors. Lung cancer is usually divided into two main groups, which account for up to about 95% of all cases. The division into groups is based on the type of cells that make up the cancer. The two main types of lung cancer are characterized by the size of the tumor cells when viewed under a microscope. They are small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). NSCLC includes several tumor subtypes. SCLC is less common, but it grows faster and metastasizes more frequently than NSCLC. Often, SCLC has already spread to other parts of the body when cancer is diagnosed. About 5% of lung cancers have rare cell types, including carcinoid tumor, lymphoma, and others. As used herein, the term "lung cancer" includes, without limitation, SCLC, NSCLC, carcinoid tumor, lymphoma, and various subtypes thereof.

Non-small cell lung cancer

NSCLC is a lung cancer that is not a type of small cell carcinoma (oat cell carcinoma). The term "non-small cell lung cancer" is applied to various types of bronchogenic carcinomas (developing from the lining of the bronchi). Examples of specific types of NSCLC include, without limitation, non-squamous cell carcinoma such as adenocarcinoma, large cell carcinoma (ie, large cell undifferentiated carcinoma), and squamous cell carcinoma.

Adenocarcinoma is a cancer that develops in the lining or inner surface of an organ. Adenocarcinoma is the most common type of lung cancer, accounting for 30%-40% of all lung cancers. A subtype of adenocarcinoma is called bronchoalveolar cell carcinoma, which creates a pneumonia-like appearance on a chest x-ray.

Squamous cell carcinoma is a cancer that begins in sloughing cells. Squamous cells are thin, flat cells that look like fish scales under a microscope. Squamous cells are found in the tissue that forms the surface of the skin, the lining of the hollow organs of the body, and the passages of the respiratory and digestive tracts. Squamous cell carcinomas can develop in any of these tissues. Squamous cell carcinoma is the second most common type of lung cancer, accounting for about 30% of all cases.

Large cell carcinoma does not show signs of squamous or glandular maturation. Consequently, such tumors are often diagnosed by exclusion when all other possibilities have been excluded. Such tumors do not have any diagnostic characteristics to suggest their diagnosis prior to biopsy. They tend to grow rapidly, metastasize early, and are clearly associated with smoking. Large cell tumors are usually large, massive, well-filled, pink-gray masses with intense hemorrhages and necrosis. Although they usually have central necrosis, they rarely cavitate. They tend to be present from the middle to the peripheral zones of the lung. They can spread locally, involving segmental or subsegmental bronchi. A variant of large cell carcinoma is giant cell carcinoma. This subtype is particularly aggressive and has a very poor prognosis. Such tumors usually present as a large peripheral mass with a focal necrotic component. They do not involve the major airways except by direct stretching. Large cell carcinoma accounts for 10%-20% of all lung cancers.

In some embodiments, the method described herein comprises administering to an NSCLC patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein said patient has elevated levels of PD-L1 in a squamous or non-squamous tumor. In some embodiments, the method described herein comprises administering to an NSCLC patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein said patient has a PD-L1 tumor proportion score (PD-L1 TPS) from 1% to 50% in a tissue sample of a squamous or non-squamous tumor. In some embodiments, the method described herein comprises administering to an NSCLC patient a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein said patient has a PD-L1 tumor proportion score (PD-L1 TPS) 50% or more in a tissue sample of a squamous or non-squamous tumor. PD-L1 TPS is a measure of the percentage of cells in a tumor tissue sample that stain positive for PD-L1 expression as determined using immunohistochemistry. In some embodiments, PD-L1 TPS is determined using the PD-L1 IHC 22C3 pharmDx kit.

Melanoma

Melanoma is a malignant tumor of melanocytes, which are cells that produce the pigment melanin and originate from the neural crest. Although most melanomas develop on the skin, they can also develop on mucous membranes or other areas to which neural crest cells migrate, including the uveal tract. Uveal melanoma differs significantly from cutaneous melanoma in incidence, prognostic factors, molecular characteristics, and treatment.

In the United States in 2014, 9,710 people were believed to have died from melanoma and the number of new cases is estimated at 76,100. Skin cancer is the most common malignancy diagnosed in the United States, with 3.5 million cancers diagnosed in 2 million people annually. Melanoma represents less than 5% of skin cancers but is responsible for the majority of deaths. The frequency has been increasing over the past four decades. Older men are at the highest risk; however, melanoma is the most common cancer in young adults aged 25 to 29 and the second most common cancer in ages 15 to 29. Ocular melanoma is the most common eye cancer, with approximately 2000 cases diagnosed annually.

Melanoma occurs predominantly in adults and more than 50% of cases develop in an apparently normal area of the skin. Although melanoma can develop anywhere, including the mucous membranes and choroid of the eye, melanoma in women develops more frequently on the extremities and in men it develops more frequently on the trunk or head and neck.

The prognosis of melanoma is influenced by the characteristics of primary and metastatic tumors. The most important prognostic factors include, without limitation: thickness and/or level of melanoma invasion, mitotic index, defined as mitoses per millimeter, ulceration or bleeding at the primary site, number of regional lymph nodes involved, distinguishing between macrometastases and micrometastases, systemic metastasis, local- non-visceral versus lung versus other visceral areas, elevated serum lactate dehydrogenase. Without being bound by any theory, it is contemplated that the presence of tumor-infiltrating lymphocytes may be a potential prognostic factor. PD-1 is reported to be predominantly expressed by tumor-infiltrating T-lymphocytes in melanoma.

Treatment options for non-small cell lung cancer and melanoma

One embodiment of the invention provides a method of treating cancer in a patient, wherein the method comprises administering to the patient a combination containing an entinostat and an anti-PD-1 or anti-PD-L1 antibody. Another embodiment of the invention provides a method of treating cancer in a patient, wherein the method comprises administering to the patient a combination containing an entinostat and an anti-PD-1 antibody. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is pembrolizumab.

Another embodiment of the invention provides a method wherein the cancer is characterized by increased expression of PD-L1. Another embodiment of the invention provides a method wherein the cancer is characterized by low expression of PD-L1. Another embodiment of the invention provides a method wherein the cancer is lung cancer or melanoma. Another embodiment of the invention provides a method wherein the cancer is lung cancer. Another embodiment of the invention provides a method wherein the lung cancer is non-small cell lung cancer. Another embodiment of the invention provides a method wherein the non-small cell lung cancer is adenocarcinoma. Another embodiment of the invention provides a method wherein the non-small cell lung cancer is squamous cell carcinoma. Another embodiment of the invention provides a method wherein the non-small cell lung cancer is a large cell carcinoma. Another embodiment of the invention provides a method wherein the cancer is melanoma.

Another embodiment of the invention provides a method wherein the patient has received at least one prior therapy. Another embodiment of the invention provides a method wherein the prior therapy is for the treatment of cancer. Another embodiment of the invention provides a method where the cancer is locally advanced. Another embodiment of the invention provides a method where the cancer is metastatic. Another embodiment of the invention provides a method wherein the prior therapy was with an anti-PD-1 antibody.

Another embodiment of the invention provides a method wherein the entinostat and anti-PD-1 or anti-PD-L1 antibodies are administered sequentially in any order or simultaneously. Another embodiment of the invention provides a method wherein the entinostat and the anti-PD-1 antibody are administered sequentially in any order or simultaneously. Another embodiment of the invention provides a method wherein the entinostat and the anti-PD-1 antibody are administered sequentially, in any order, or simultaneously, during a 21 day treatment cycle. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered on day 1 of the treatment cycle. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 0.1-10 mg/kg. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered as an intravenous infusion. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered as an intravenous infusion over a period of 15 minutes to 60 minutes. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg as an intravenous infusion over a period of 15 minutes to 60 minutes. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg as an intravenous infusion over a period of 30 minutes. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg as an intravenous infusion over a period of 15 minutes to 60 minutes once every 2 weeks. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg as an intravenous infusion over a period of 15 minutes to 60 minutes once every 3 weeks. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a dose of 2 mg/kg as an intravenous infusion over a period of 15 minutes to 60 minutes once every 4 weeks. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a fixed dose of 200 mg. Another embodiment of the invention provides a method wherein the anti-PD-1 antibody is administered at a fixed dose of 200 mg, every 3 weeks, via intravenous infusion. Another embodiment of the invention provides a method wherein the entinostat is administered intermittently during a treatment cycle. Another embodiment of the invention provides a method wherein the entinostat is administered on day 1 of the treatment cycle. Another embodiment of the invention provides a method wherein the entinostat is administered on day 15 of a treatment cycle. Another embodiment of the invention provides a method wherein the entinostat is administered at a dose of 3 mg/kg. Another embodiment of the invention provides a method wherein the entinostat is administered at a dose of 5 mg/kg. Another embodiment of the invention provides a method wherein the entinostat is administered at a dose of 10 mg/kg. Another embodiment of the invention provides a method where the entinostat is administered first. Another embodiment of the invention provides a method where the entinostat is administered intermittently. Another embodiment of the invention provides a method wherein the entinostat is administered weekly. Another embodiment of the invention provides a method wherein the entinostat is administered biweekly. Another embodiment of the invention provides a method wherein the entinostat is administered biweekly, at a dose of 10 mg/kg. Another embodiment of the invention provides a method wherein an entinostat and an anti-PD-1 or anti-PD-L1 antibody are administered simultaneously. Another embodiment of the invention provides a method wherein an entinostat and an anti-PD-1 or anti-PD-L1 antibody are administered simultaneously.

In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include any additional therapeutic agent. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include any additional immune checkpoint inhibitor. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not contain an anti-CTLA4 inhibitor. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination containing an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include 5-azacytidine.

In one embodiment, a method of treating cancer in a patient is provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the entinostat is provided in a solid dosage form. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the anti-PD-1 antibody or anti-PD-L1 antibody given as an intravenous infusion. In some embodiments, methods are provided for treating cancer in a patient comprising administering to the patient a combination comprising essentially an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the entinostat is provided in a solid dosage form and an anti-PD-1 antibody, or anti-PD-L1 antibody is provided as an intravenous infusion. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting predominantly of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the entinostat is provided in a solid dosage form and the anti-PD-1 antibody is provided. in the form of intravenous infusion. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of entinostat and pembrolizumab, wherein the entinostat is provided in a solid dosage form and pembrolizumab is provided in the form of an intravenous infusion.

In one embodiment, a method of treating cancer in a patient is provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include any additional therapeutic agent. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include any additional immune checkpoint inhibitor. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include an anti-CTLA4 inhibitor. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the combination does not include 5-azacytidine.

In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and pembrolizumab. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of an entinostat and pembrolizumab, wherein said combination does not include an anti-CTLA4 inhibitor. In some embodiments, methods of treating cancer in a patient are provided, comprising administering to the patient a combination consisting essentially of entinostat and pembrolizumab, wherein said combination does not include 5-azacytidine.

Ways to select patients for combination therapy

Provided herein is a method for selecting patients for combination therapy involving the administration of an entinostat and an anti-PD-L1 antibody, where the selection is based on the level of PD-L1 expression in the tumor.

Provided herein in a further embodiment is a method for selecting patients for combination therapy comprising administering an entinostat and an anti-PD-1 or anti-PD-L1 antibody, where selection is based on tumor proportion score (TPS) for PD-L1 expression. The tumor proportion score is an estimate of the percentage of cells in a tumor tissue sample that stain positive for PD-L1 expression, as determined using immunohistochemistry. In some embodiments of the invention, TPS is determined using the PD-L1 IHC 22C3 pharmDx kit.

In some embodiments, the method further comprises administering a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody to patients expressing high levels of PD-L1 in a tumor. In some embodiments, the method further comprises administering a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody to patients wherein the TPS is between 1% and 50%. In some embodiments, the method further comprises administering a combination therapy comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody to patients where the TPS is greater than or equal to 50%. In some embodiments, the tumor tissue sample where PD-L1 expression is measured is obtained from a patient with non-small cell lung cancer. In some embodiments, the non-small cell lung cancer is a squamous cell carcinoma or a non-squamous cell carcinoma. In some embodiments, the tumor sample where PD-L1 expression is measured is obtained from a patient with melanoma. In some embodiments, the dosage of the anti-PD-L1 or anti-PD-L1 antibody used in combination therapy is determined based on the expression of PD-L1 in tumor samples.

Methods for assessing patients during the course of treatment

In a further embodiment, the present invention provides a method for treating cancer in a patient by administering a combination comprising an entinostat and an anti-PD-1 or anti-PD-L1 antibody, wherein the method further comprises the steps of evaluating biomarkers in a biological sample obtained from the patient during a course of combined therapy. In some embodiments of the invention, the biological sample is a blood sample. In some embodiments, the biomarker is Lin (CD3, CD14, CD19, CD56) negative, HLA-DR-, CD33+ myeloid-derived suppressor cells (MDSC). In some embodiments, the biomarker is CD11b+CD14-CD33+ MDSC. In some embodiments, the biomarker is CD11b+; CD14-; CD33+; CD15+ polymorphonuclear-MDSC. In some embodiments, the biomarker is CD11b+; CD14-; CD33+; CD15- monocytic MDSC. In some embodiments, the biomarker is CD14+; HLA-DR-/lo monocytic MDSC. In some embodiments, the biomarker is CD14+CD16+CD66b-HLA-DR+ monocytes, including subgroups of classic CD14hiCD16, intermediate CD14hiCD16+, and non-classical CD14+CD16hi. In some embodiments, the biomarker is Lin-HLA-DR+CD303+ (BDCA2) plasmacytoid dendritic cells. In some embodiments, the biomarker is Lin-HLA-DR+CD1c+ (BDCA1) myeloid dendritic cells. In some embodiments, the biomarker is Lin-HLA-DR+CD141+ (BDCA3) dendritic cells. In some embodiments, the biomarker is CD11b+CD14-CD15+/CD66b+ neutrophil cells in the high density fraction. In some embodiments, the biomarker is CD3+CD4+; CD3+CD8+ T cells. In some embodiments, the biomarker is CD4+CD25+FoxP3+ regulatory T cells. In some embodiments, the biomarker is CD19+ B cells.

In some embodiments of the invention, the introduction of a combination therapy, including entinostat and anti-PD-1 or anti-PD-L1 therapy, leads to a decrease in MDSC cells in a peripheral blood sample. In some embodiments of the invention, a decrease in MDSC cells in a peripheral blood sample during a course of treatment correlates with a positive result of combination therapy.

In some embodiments, the biomarker is the level of lysine protein acetylation in the tumor sample. In some embodiments, the biomarker is the level of expression of PD-L1 in a tumor sample. In some embodiments, the biomarker is a tumor proportion score (TPS) for PD-L1 expression. In some embodiments, TPS was determined using the PD-L1 IHC 22C3 pharmDx kit. In some embodiments, the biomarker is the level of e-cadherin in the tumor sample. In some embodiments, the biomarker is CD33+ and S100A9+ myeloid-derived suppressor cells (MDSC) in the tumor sample. In some embodiments, the biomarker is CD33 ⁺ and S100A9 ⁺ myeloid-derived suppressor cells (MDSC) in a peripheral blood sample. In some embodiments, the biomarker is CD163 ⁺ or CD68 ⁺ macrophages in a tumor sample. In some embodiments, the biomarker is CD163 ⁺ or CD68 ⁺ macrophages in a peripheral blood sample. In some embodiments, the biomarker is neutrophil elastase in a tumor sample. In some embodiments, the biomarker is neutrophil elastase in a serum sample. In some embodiments, the biomarker is dendritic cells (DC) expressing type II of the C-type lectin DC-SIGN (CD209). In some embodiments, DC-SIGN (CD209) is determined in serum. In some embodiments, DC-SIGN (CD209) is determined in a tumor sample. In some embodiments, the biomarker is CD4, CD8, Granzyme B, and FoxP3 positive cells in tumor samples.

Complementary Therapy

Available complementary treatments for lung cancer and melanoma that can advantageously be used in combination with the therapy described herein include, but are not limited to, radiation therapy, chemotherapy, antibody therapy, and tyrosine kinase inhibitor therapy as adjuvant therapy.

Radiation therapy is a cancer treatment that uses high-energy x-rays or other types of radiation to kill cancer cells or stop them from growing. Chemotherapy is a cancer treatment that uses drugs to stop the growth of cancer cells, either by killing the cells or by stopping them from dividing. When chemotherapy is taken by mouth or injected into a vein or muscle, the drugs enter the bloodstream and can reach cancer cells throughout the body (systemic chemotherapy). When chemotherapy is placed in the spinal cord, an organ, or a body cavity such as the abdomen, the drugs primarily target cancer cells in that area (regional chemotherapy). The way chemotherapy is given depends on the type and stage of cancer being treated.

Various chemotherapeutic agents are known in the art for the treatment of lung cancer. Cytotoxic agents used to treat lung cancer include carboplatin (eg, Paraplatin®, Paraplat®), cisplatin (eg, Platinol®, Platinol-Aq®), crizotinib (eg, Xalkori®), etoposide (eg, Toposar®, VePezid® ), etoposide phosphate (eg, Etopophos®), gemcitabine hydrochloride (eg, Gemzar®), gemcitabine-cisplatin, methotrexate (eg, Abitrexate®, Folex®, Folex Pfs®, Methotrexate Lpf®, Mexat®, Mexat-Aq®) , paclitaxel (eg Taxol®), pemetrexed disodium (eg Alimta®), and topotecan hydrochloride (eg Hycamtin®)

Various agents are known in the art for the treatment of melanoma, including aldesleukin (eg Proleukin®), dabrafenib (eg Tafinlar®), dacarbazine (eg DTIC-Dome®), recombinant interferon alfa-2b (eg Intron® A), Ipilimumab (eg, Yervoy®), pembrolizumab (eg, Keytruda®), Trametinib (eg, Mekinist®), Nivolumab (eg, Opdivo®), Peginterferon alfa-2b (eg, Pegintron®, Silatron®), vemurafenib (eg, Zelboraf®).

Monoclonal antibody therapy is a cancer treatment that uses antibodies made in a laboratory from a single cell type of the immune system. Such antibodies can identify substances on cancer cells or normal substances that can help cancer cells grow. Antibodies attach to substances and destroy cancer cells, block their growth or prevent them from spreading. Monoclonal antibodies are administered by infusion. They can be used alone or carry drugs, toxins, or radioactive substances directly into cancer cells. Monoclonal antibodies are also used in combination with chemotherapy as adjuvant therapy.

Additionally, illustratively, treatments that may advantageously be combined with the compositions and therapies described herein may include, without limitation, administering agents including, but not limited to, lapatinib alone or in combination with capecitabine, docetaxel, epirubicin, epothilone A , B or D, goserelin acetate, paclitaxell, pamidronate, bevacizumab, or trastuzumab.

In some embodiments, the adjunctive therapy comprises administering to the patient one or more of doxorubicin, cyclophosphamide, paclitaxel, lapatinib, capecitabine, trastuzumab, bevacizumab, gemcitabine, eribulin, or nab-paclitaxel.

Oral compositions

Oral compositions containing the active pharmaceutical ingredients described herein may include any commonly used oral form, including: tablets, capsules, pills, lozenges, dragees, lozenges, cachets, pellets, medicated chewing gum, granules, bulk powder, effervescent or non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers, sprinkles, elixirs, syrups, buccal forms and oral liquids. Capsules may contain mixtures of the active compound(s) with inert excipients and/or diluents such as pharmaceutically acceptable starches (e.g. corn, potato or tapioca starch), sugars, artificial sweeteners, powdered cellulose such as crystalline and microcrystalline cellulose, flour, gelatins, gums, etc. Useful tablet compositions can be prepared by conventional compression, wet granulation, or dry granulation methods and use pharmaceutically acceptable diluents, astringents, lubricants, disintegrants, surface modifying agents (including surfactants), suspending agents, or stabilizing agents, including, without limitation, magnesium stearate, stearic acid, talc, sodium lauryl sulfate, microcrystalline cellulose, calcium carboxymethylcellulose, polyvinylpyrrolidone, gelatin, alginic acid, gum arabic, xanthan gum, sodium citrate, complex silicates, calcium carbonate, glycine, dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, talc, dry starches and powdered sugar. In some embodiments of the invention, surface modifying agents are present, which include nonionic and anionic surface modifying agents. For example, surface modifying agents include, without limitation, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitol esters, colloidal silicon dioxide, phosphates, sodium dodecyl sulfate, magnesium aluminum silicate, and triethanolamine. The oral compositions herein may use standard delayed or time-release formulations to alter absorption of the active compound(s). The oral composition may also consist of administering the active ingredient in water or fruit juice containing appropriate diluents or emulsifiers, as appropriate.

Oral administration

As described herein, the combination therapy described herein may be given simultaneously, or may be given in alternating fashion, with an entinostat given at a different time during a course of chemotherapy than an EGFR inhibitor. Such time difference can vary from minutes, hours, days, weeks or more between administrations of the compounds. Therefore, the term combination does not necessarily mean administered at the same time or as a single dose, but each of the components is administered during the desired period of treatment. Funds can be entered in various ways. As is typical for chemotherapeutic regimens, the course of chemotherapy may be repeated after a few weeks, and may follow the same time frame for the administration of the two compounds, or may be modified based on the patient's response.

In other embodiments, the pharmaceutical compositions provided herein may be presented in solid, semi-solid, or liquid dosage forms for oral administration. As used herein, oral administration also includes buccal, lingual, and sublingual administration. Suitable oral dosage forms include, without limitation, tablets, capsules, pills, lozenges, dragees, lozenges, cachets, pellets, medicated chewing gum, granules, bulk powder, effervescent and non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers , sprinklers, elixirs and syrups. In addition to the active ingredient(s), pharmaceutical compositions may contain one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, astringents, excipients, diluents, disintegrants, wetting agents, lubricants, glidants, colorants, dye migration inhibitors , sweeteners and flavors.

Astringents or granulators impart integrity to the tablet to ensure that the tablet remains intact after compression. Suitable binders or granulators include, without limitation, starches such as corn starch, potato starch and peptized starch (eg STARCH 1500); gelatin; sugars such as sucrose, glucose, dextrose, molasses and lactose; natural and synthetic gums such as gum arabic, alginic acid, alginates, Irish moss extract, Panwar gum, ghatti gum, isabgol snail slime, carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone (PVP), veegum, larch arabogolactan, powdered tragacanth, and guar gum; celluloses such as ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose, methylcellulose, hydroxyethylcellulose (HEC), hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC); microcrystalline celluloses such as AVICEL-PH-101, AVICEL-PH-103, AVICEL RC-581, AVICEL-PH-105 (FMC Corp., Marcus Hook, PA); and their mixtures. Suitable fillers include, without limitation, talc, calcium carbonate, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, peptized starch, and mixtures thereof. The binder or excipient may be present in an amount of from about 50 to about 99 wt % in the pharmaceutical compositions described herein.

Suitable diluents include, without limitation, dicalcium phosphate, calcium sulfate, lactose, sorbitol, sucrose, inositol, cellulose, kaolin, mannitol, sodium chloride, dry starch, and powdered sugar. Certain diluents such as mannitol, lactose, sorbitol, sucrose, and inositol, when present in sufficient amounts, can interfere with the properties of some compressed tablets to allow disintegration in the mouth upon chewing. Such compressed tablets may be used as chewable tablets.

Suitable disintegrants include, without limitation, agar; bentonite; celluloses such as methylcellulose and carboxymethylcellulose; wood products; natural sponge; cation exchange resins; alginic acid; gums such as guar gum and veegum HV; citrus pulp; cross-linked celluloses such as croscarmellose; cross-linked polymers such as crospovidone; cross-linked starches; calcium carbonate; microcrystalline cellulose such as sodium starch glycolate; potassium polacrilin; starches such as corn starch, potato starch, tapioca starch and peptized starch; clay; equalizers; and their mixtures. The amount of disintegrating agent in the pharmaceutical compositions provided herein varies with the type of composition, and is obvious to one of ordinary skill in the art. The pharmaceutical compositions provided herein may contain from about 0.5% to about 15%, or from about 1% to about 5%, by weight of a disintegrant.

Suitable lubricants include, without limitation, calcium stearate; magnesium stearate; mineral oil; light mineral oil; glycerol; sorbitol; mannitol; glycols such as glycerol behenate and polyethylene glycol (PEG); stearic acid; sodium lauryl sulfate; talc; hydrogenated vegetable oil, including peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil; zinc stearate; ethyl oleate; ethyl laureate; agar; starch; lycopodium; silica or silica gels such as AEROSIL® 200 (W.R. Grace Co., Baltimore, MD) and CAB-O-SIL® (Cabot Co. of Boston, MA); and their mixtures. Pharmaceutical compositions provided herein may contain about 0.1 to about 5 wt% lubricant.

Suitable glidants include colloidal silica, CAB-O-SIL® (Cabot Co. of Boston, MA), and non-asbestos talc. The colorants include any of FD&C approved, certified, water-soluble colorants, and FD&C water-insoluble colorants suspended in aluminum hydrate, and lacquers, and mixtures thereof. Lacquer is a combination of absorption of a water-soluble paint on an aqueous heavy metal oxide, resulting in an insoluble form of the dye. Flavors include natural flavors extracted from plants, such as fruits, and synthetic blends of compounds that give a pleasant taste sensation, such as peppermint and methyl salicylate. Sweeteners include sucrose, lactose, mannitol, syrups, glycerin, and artificial sweeteners such as saccharin and aspartame. Suitable emulsifiers include gelatin, gum arabic, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitol monooleate (TWEEN® 20), polyoxyethylene sorbitol monooleate 80 (TWEEN® 80), and triethanolamine oleate. Suspending and dispersing agents include sodium carboxymethylcellulose, pectin, tragacanth, veegum, gum arabic, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Preservatives include glycerin, methyl and propyl paraben, benzoic acid, sodium benzoate, and alcohol. Moisturizers include propylene glycol monostearate, sorbitol monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether. Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examples of non-aqueous liquids used in emulsions include mineral oil and cottonseed oil. Organic acids include citric and tartaric acid. Sources of silica include sodium bicarbonate and sodium carbonate.

It is to be understood that a plurality of carriers and excipients can perform multiple functions, even in a single composition.

In further embodiments, the pharmaceutical compositions provided herein may be provided as compressed tablets, compressible tablets, chewable lozenges, fast dissolving tablets, multiple compressed tablets, or enteric, sugar, or film-coated tablets. Enteric-coated tablets are compressed tablets containing substances that resist the action of gastric acid but dissolve or break down in the intestine, thus protecting the active ingredients from the acidic environment of the stomach. Enteric coatings include, without limitation, fatty acids, fats, phenyl salicylate, waxes, shellac, ammoniated shellac, and cellulose acetate phthalates. Sugar-coated tablets are compressed tablets surrounded by a sugar shell, which can be useful in covering unpleasant tastes or odors and in protecting the tablets from oxidation. Film-coated tablets are compressed tablets that are coated with a thin layer or film of a water-soluble material. Film shells include, without limitation, hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Film casings impart the same general characteristics as sugar casings. Multi-compressed tablets are compressed tablets made by more than one compression cycle, including layered tablets and press-coated or dry-coated tablets.

Tablet dosage forms may be prepared from the active ingredient in powder, crystalline, or granular forms, alone or in combination with one or more of the carriers or excipients described herein, including astringents, disintegrants, release-controlling polymers, lubricants, diluents and/or dyes. Flavorings and sweeteners are particularly useful in making chewable tablets and lozenges.

The pharmaceutical compositions provided herein may be in the form of soft or hard capsules, which may be made from gelatin, methylcellulose, starch, or calcium alginate. A hard gelatin capsule, also known as a dry-filled capsule (DFC), consists of two compartments, one on top of the other, thus completely covering the active ingredient. A soft elastic capsule (SEC) is a soft globular shell, such as a gelatin shell, that is plasticized by the addition of glycerol, sorbitol, or a similar polyol. Soft gelatin capsules may contain a preservative to prevent the growth of microorganisms. Suitable preservatives are those described herein, including methyl and propyl parabens, and sorbic acid. Liquid, semi-solid, and solid dosage forms provided herein may be encapsulated in a capsule. Suitable liquid and semi-solid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils or triglycerides. Capsules containing such solutions can be prepared as described in US Pat. 4328245; 4409239; and 4,410,545. Capsules may also be coated, as is known to those skilled in the art, to modify or maintain dissolution of the active ingredient.

In other embodiments, the pharmaceutical compositions described herein may be provided in liquid and semi-solid dosage forms, including emulsions, solutions, suspensions, elixirs, and syrups. An emulsion is a two-phase system in which one liquid is dispersed in the form of small globules throughout another liquid, which may be oil-in-water or water-in-oil. Emulsions may include pharmaceutically acceptable non-aqueous liquids or a solvent, an emulsifying agent, and a preservative. Suspensions may include a pharmaceutically acceptable suspending agent and a preservative. Aqueous-alcoholic solutions may include a pharmaceutically acceptable acetal such as a lower alkyl aldehyde di(lower alkyl) acetal (the term "lower" means alkyl having 1 to 6 carbon atoms), for example, acetaldehyde diethyl acetal; and a water-miscible solvent having one or more hydroxyl groups such as propylene glycol and ethanol. Elixirs are clear, sweet and water-alcohol solutions. Syrups are concentrated aqueous solutions of sugar, such as sucrose, and may also contain a preservative. For a liquid dosage form, for example, the polyethylene glycol solution may be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier, for example, water, for easy metering upon administration.

Other useful liquid and semi-solid dosage forms include, without limitation, those containing the active ingredient(s) provided herein and dialkylated mono- or polyalkylene glycol, including, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350 -dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether, where 350, 550, and 750 refer to the approximate average molecular weight of polyethylene glycol. Such compositions may further include one or more antioxidants such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, bisulfite, sodium metabisulphite, thiodipropionic acid and its esters and dithiocarbamates.

Pharmaceutical compositions provided herein for oral administration may also be provided in the form of liposomes, micelles, microspheres, or nanosystems. Micellar dosage forms can be prepared as described in US Patent No. 6350458.

In other embodiments, the pharmaceutical compositions provided herein may be presented as non-effervescent or effervescent granules or powders, for reconstitution into a liquid dosage form. Pharmaceutically acceptable carriers and excipients used in non-effervescent granules or powders may include diluents, sweeteners and wetting agents. Pharmaceutically acceptable carriers and excipients used in effervescent granules or powders may include organic acids and a source of carbon dioxide.

Dyes and flavors can be used in all of the above dosage forms.

The pharmaceutical compositions provided herein may be formulated as immediate or modified release dosage forms, including delayed, sustained, pulsed, controlled, targeted, and programmed release forms.

In additional embodiments, the pharmaceutical compositions provided herein may be co-formulated with other active ingredients that do not interfere with the desired therapeutic effect, or with substances that complement the desired effect.

EXAMPLES

Example 1

An open-label phase 1B/2 escalation study of entinostat in combination with pembrolizumab in patients with non-small cell lung cancer, with an expansion cohort in patients with non-small cell lung cancer and melanoma.

The combination of entinostat with pembrolizumab improves response to PD-1 blocking antibody treatment and results in an improved response rate compared to administration of each agent alone in patients with non-small cell lung cancer and melanoma, with an acceptable safety profile.

The main objective:

- Number of participants receiving 3 mg entinostat per week with adverse events as a measure of safety and tolerability.

- Number of participants receiving 5 mg entinostat per week with adverse events as a measure of safety and tolerability.

- Number of participants receiving 10 mg entinostat per week with adverse events as a measure of safety and tolerability.

Phase 1b (Dose Upscaling/Confirmation Cohorts):

- to determine the dose-limiting toxicity (DLT) and maximum tolerated dose (MTD) or dose recommended for phase 2 (RP2D) of entinostat (SNDX-275) given in combination with pembrolizumab.

- Phase 2 (expanded cohorts): To assess the preliminary efficacy of entinostat in RP2D in combination with pembrolizumab in patients with melanoma and NSCLC, as determined by the overall response rate (ORR), according to the criteria for evaluating the immune response in solid tumors (irRECIST) in each evaluated cohort.

Additional goals:

Efficacy: To evaluate the efficacy of entinostat in combination with pembrolizumab in patients with melanoma and NSCLC, as determined by secondary efficacy evaluations based on irRECIST and RECIST 1.1, including:

- clinical benefit rate (CBR) (i.e. complete response [CR]+partial response [PR]+stable disease [SD]) at 6 months.

- progression-free survival (PFS) status at 6 months.

- PFS

- overall survival (OS)

• in patients with a response to treatment (ie CR or PR):

• response time (DOR)

• time to response (TTR)

Safety: To evaluate the safety and tolerability of entinostat in combination with pembrolizumab as assessed by clinical adverse events (AE), laboratory parameters and electrocardiograms (ECG). Study design

Patients included in the combination therapy study described herein received two weeks of entinostat monotherapy alone. After completion of two weeks of monotherapy, patients entered the treatment cycle of the combination therapy period. Each treatment cycle lasted 21 days, during which patients received entinostat in combination with pembrolizumab. Phase 2 (Expansion phase)

the main objective

To evaluate the preliminary efficacy of entinostat in RP2D in combination with pembrolizumab in patients with melanoma and NSCLC, as determined previously by overall response rate (ORR).

Additional goals

Additional targets are the same as those listed for Phase 1B.

Key Endpoints

- overall response rate (ORR)

- clinical benefit level (CBR)

- duration of response (DOR)

- time to response (TTR)

- progression-free survival (PFS) status at 6 months

- progression-free survival (PFS)

- overall survival (OS)

Additional endpoints

Safety: AEs, clinical laboratory parameters, and ECG.

Pharmacodynamics: Protein acetylation in peripheral blood mononuclear cells (PBMC)

Research:

- Changes in expression of checkpoint inhibitor (PD-1/PD-L1) in tumor biopsies before and after therapy

- ratio of effector T cells:regulatory T cells in tumor biopsies before and after therapy (immunohistochemistry)

- changes in the amount of MDSC in peripheral blood and tumor biopsies

- changes in protein lysine acetylation in peripheral blood cells and tumor biopsies before and after treatment

- PK profiles of entinostat and pembrolizumab

- changes in the specialization of inflammatory T cells in the blood and tumor biopsies before and after therapy

- gene expression profile before and after biopsy

- changes in other immune markers of regulatory mechanisms and other evaluations of received/stored samples of biological samples

- Immunogenicity of pembrolizumab in combination with entinostat

- entinostat exposure-safety response when administered in combination with pembrolizumab

The blood levels of the following exemplary immunological efficacy markers were assessed:

- suppressor cells of myeloid origin (MDSC): Lin (CD3, CD14, CD19, CD56) negative; HLA-DR-; CD33 ⁺

- MDSC: CD11b ⁺ CD14 ^- CD33 ⁺

- polymorphonuclear-MDSC (PMN-MDSC): CD11b ⁺ ; ^CD14- ; CD33 ⁺ ; CD15 ⁺

- monocytic (M-MDSC): CD11b ⁺ ; ^CD14- ; CD33 ⁺ ; ^CD15-

- M-MDSC: CD14 ⁺ ; HLA-DR ^-/lo

- monocytes: CD14 ⁺ CD16 ⁺ CD66b ^- HLA ^- DR ⁺ , including subgroups of classic CD14 ^hi CD16 ⁺ , intermediate CD14 ^hi CD16 ⁺ , and non-classical CD14 ⁺ CD16 ^hi

- dendritic cells: Lin ^- HLA ^- DR ⁺ CD303 ⁺ (BDCA2) plasmacytoid; Lin ^- HLA ^- DR ⁺ CD1c ⁺ (BDCA1) myeloid; Lin ^- HLA ^- DR ⁺ CD141 ⁺ (BDCA3)

- neutrophils: CD11b ⁺ CD14 ^- CD15 ⁺ /CD66b ⁺ cells in the high density fraction

- T cells: CD3 ⁺ CD4 ⁺ ; CD3 ⁺ CD8 ⁺ ; regulatory T cells: CD4 ⁺ CD25 ⁺ FoxP3 ⁺

- B cells: CD19

Cellular functions were assessed by measuring levels of interleukin-2 (IL-2), interferon gamma (IFN-γ), and granulocyte-macrophage-colony stimulating factor (GM-CSF) in mononuclear cells stimulated with CD3/CD28, ConA, and tetanus toxin.

Cell proliferation was assessed by the ³ H-thymidine consumption assay.

Tumor biopsy tissues obtained at various stages of treatment with the combination therapy described herein were evaluated for:

- acetylation

- PD-L1 expression

- e-cadherin levels

- MDSC: CD33 ⁺ S100A9 ⁺

- macrophage cells CD163 ⁺ or CD68 ⁺

- neutrophils - neutrophil-elastase cells ⁺

- dendritic cells - DC-SIGN (CD209)

- CD4, CD8, granzyme B and FoxP3 positive cells, if appropriate, if the level of MDSC in the tissue is reduced

Study design

The expansion phase was started after DLT, MTD and RP2D were determined during the dose escalation phase. In this phase, the entinostat was evaluated using the RP2D identified in the dose escalation phase. At baseline, patients will receive entinostat at an initial dose of 3 mg at D1, D8, and D15 along with pembrolizumab 200 mg by intravenous (IV) infusion at D1 of a 21-day cycle.

The following patient cohorts may be selected to participate in this phase:

- cohort 1 : NSCLC, with squamous cell tumor or adenocarcinoma histology, who were not treated with PD-1 or PDL-1 blocking antibodies

- cohort 2 : patients with NSCLC (any histology) who were previously treated and progressed on PD-1 or PD-L1-blocking antibodies

- cohort 3 : patients with melanoma who had previously received and progressed on PD-1 or PD-L1-blocking antibodies.

Entinostat monotherapy immunocorrelated cohort (EMIC) : Up to 20 patients with stage 2 NSCLC cohort 1 will be randomly selected to participate. Sample size

Dose escalation/confirmation phase

Three to 6 patients will be included in each dose cohort based on the standard phase 1 dose escalation schedule. Each patient will only participate in the 1 dose cohort. The total number of patients who will be included in the dose escalation/confirmation phase depends on the observed safety profile, which will determine the number of patients per dose cohort, as well as the number of dose escalations required to achieve the MTD or RP2D. A total of 9 additional patients will be included in the potential RP2D dose confirmation cohort to obtain additional AEs, immune correlation and antitumor activity data on entinostat at MTD or another dose recommended for further study in phase 2 (i.e. RP2D) in combination.

Expansion phase

It is planned to include up to 136 patients in 3 cohorts. Patients will be included in each cohort according to the design of an uncontrolled study with ORR as the primary endpoint. The expansion phase will be carried out in 2 stages, so that the inclusion of 1 or more of the cohorts being evaluated can end early in case the antitumor activity of the combination regimen is insufficient. The decision to end or continue enrollment for each cohort will be made independently of other cohorts. The protocol may be modified to allow inclusion of additional or other cohorts based on data obtained during the course of the study. Combination therapy with entinostat and pembrolizumab will be evaluated in each of the four cohorts, following the criteria described below:

Inclusion Criteria

Patients with NSCLC :

1. Have histologically or pathologically confirmed recurrent/metastatic NSCLC.

2. In the presence of adenocarcinoma, prior testing for anaplastic lymphoma kinase (ALK) rearrangement and epidermal growth factor receptor (EGFR) mutations is required, with available results in this study, and, if positive, prior treatment with EGFR or ALK.

3. Received at least 1 chemotherapeutic regimen under conditions of spread/metastasis

Melanoma patients:

1. Have a histologically or cytologically confirmed diagnosis of unresectable or metastatic melanoma and have experienced PD after a PD-1 or PD-L1 blocking antibody and, if BRAF V600 positive, a BRAF inhibitor. Patients in expansion phase, cohorts 2 and 3

2. Previously received PD-1/PD-L1 blocking antibody treatment and developed PD during such treatment.

Patients in expansion phase, cohorts 2 (NSCLC) and 3 (melanoma)

1. Previously treated with a PD-1/PD-L1 blocking antibody (i.e. pembrolizumab, nivolumab, MEDI4736, or GNE PD-L1 [MPDL3280A]) and experienced documented unequivocal progressive disease according to RECIST 1.1 or clinically during or after such treatment.

All patients :

1. Age 18 years or older on the date of signing the informed consent.

2. In the presence of brain metastases, must have a stable neurological status after topical therapy for at least 4 weeks without the use of steroids or at a stable or reduced dose of ≤10 mg per day of prednisone (or equivalent), and must not have neurological dysfunction , which distorts the assessment of neurological and other AEs.

3. Evidence of locally advanced or metastatic disease, based on imaging studies within 28 days prior to the first dose of study drug:

4. At least 1 measurable lesion ≥20 mm on conventional techniques or ≥10 mm on helical CT scan or MRI, with last imaging within 28 days prior to the first dose of study drug. If there is only 1 measurable lesion and it is located in a previously irradiated area, progression must be demonstrated according to RECIST version 1.1.

5. When receiving radiation therapy, have a 2-week washout period after completion of treatment prior to receiving the first dose of study drug and continue to have at least 1 measurable lesion according to the above criteria.

6. Functional status ECOG 0 or 1.

7. Have acceptable applicable laboratory parameters.

8. Female patients must not be pregnant.

9. If male, consent to use an adequate method of contraception

10. There is a resolution of the toxic effect(s), most pronounced for previous chemotherapy to grade 1 or less (with the exception of alopecia). If a patient has received major surgery or radiation therapy >30 Gy, they should recover from toxicity and/or complications from the intervention.

11. Desire to obtain a fresh tumor sample at screening and other time points designated as mandatory, as per the examination procedure schedule.

12. Are able to understand and give written informed consent and follow research procedures.

Exclusion Criteria

Patients who meet any of the following criteria are not considered eligible to participate in the study:

1. Diagnosis of immunodeficiency or receipt of systemic steroid therapy or any other form of immunosuppressive therapy within 7 days prior to the first dose of study drug. The use of physiological doses of corticosteroids may be approved after consultation with the Sponsor.

2. Active autoimmune disease that has required systemic treatment in the last 2 years (i.e., disease-modifying agents, corticosteroids, and immunosuppressive agents). Replacement therapy (eg, thyroxine, insulin, or physiologic corticosteroid replacement therapy for adrenal or pituitary insufficiency) is not considered a form of systemic treatment.

3. History of interstitial lung disease (ILD).

4. Allergy to benzamide or inactive components of entinostat.

5. History of allergy to any active or inactive ingredients of pembrolizumab.

6. History or existing evidence of any condition, therapy, or laboratory finding that may affect the results of the study, interfere with the patient's participation in the study for the duration of the study, or is not in the patient's best interest to participate, in the opinion of the Investigator, including, without limitation:

7. Myocardial infarction or thromboembolism within 6 months from baseline, or severe or unstable angina, New York Heart Association (NYHA) class III or IV disease, or QTc interval > 470 msec.

8. Uncontrolled heart failure or hypertension, uncontrolled diabetes mellitus, or uncontrolled systemic infection.

9. Other known additional malignancy that is progressing or requires active treatment (excluding adequately treated basal cell carcinoma or cervical intraepithelial neoplasia [CIN]/cervical carcinoma in situ or melanoma in situ). Prior history of other cancer is acceptable if there has been no active disease in the previous 5 years.

10. Signs of pneumonitis or history of pneumonitis.

11. Active infection requiring systemic therapy.

12. Known active metastases to the central nervous system (CNS) and/or carcinomatous meningitis.

13. Note: Patients with previously treated brain metastases may participate, provided they are stable (no evidence of progression on imaging [using identical imaging options for each assessment, or MRI or CT scan] for at least 4 weeks prior to the first dose of study drug and any neurologic symptoms have returned to baseline), have no evidence of new or increasing brain metastases, and have not used steroids for at least 2 weeks prior to the first dose of study drug, or are on a stable or reduced dose ≤10 mg per day of prednisone (or equivalent). This exclusion does not include carcinomatous meningitis, which is excluded regardless of clinical stability.

14. Known psychiatric disorders or drug dependence that interfere with the interaction with study requirements.

15. Currently participating and on study therapy or who participated in a study agent study and received study therapy or using study device within 4 weeks of the first dose of treatment.

16. Received a live vaccine within 30 days of the first dose of treatment.

17. Previous cancer monoclonal antibodies (mAbs) within 4 weeks prior to baseline or who did not recover (i.e., ≤grade 1 or baseline) from AEs due to agents administered more than 4 weeks earlier.

18. Prior chemotherapy, targeted small molecule therapy, or radiation therapy within 2 weeks of the study at baseline or who did not recover (i.e., ≤ Grade 1 or baseline) from AEs due to the previously administered agent. Note: Patients with grade ≤ 2 neuropathy or ≤ 2 alopecia are exceptions to this criterion and may be considered for study. Note that if a patient has undergone major surgery, they should have adequately recovered from the toxicity and/or complications of the intervention prior to initiating therapy.

19. Received transfusion of blood components (including platelets or red blood cells) or administration of colony-stimulating factors (including granulocyte colony-stimulating factor [G-CSF], granulocyte-macrophage colony-stimulating factor [GM-CSF], or recombinant erythropoietin) within 4 weeks prior to the first dose of treatment .

20. Currently receiving treatment with any other agent on the Prohibited List, such as valproic acid or other systemic cancer agents, within 14 days prior to the first dose of treatment.

21. If the woman is pregnant, nursing, or expecting to conceive, or if the man is expecting to have children within the intended study period from the screening visit up to 120 days after the last dose of study drug.

22. Known history of human immunodeficiency virus (HIV) (HIV 1/2 antibodies).

23. Known active hepatitis B (eg, hepatitis B surface antigen reaction) or hepatitis C (eg, hepatitis C virus ribonucleic acid [quantitative]).

24. Is or has a close family member (eg, spouse, parent/legal guardian, sibling, or child) who works at a study center or sponsor directly involved in said study, except when approved by the Institution's Prospective Review Board (IRB). )/Ethics Committee (EC) (chair or representative) for the possibility of exclusion under this criterion for a particular patient.

Example 2

ENCORE 601: A phase 1b/2 study designed to study entinostat in combination with pembrolizumab in patients with advanced NSCLC or melanoma.

Methods

Patients with stage III/IV NSCLC previously treated with platinum-based chemotherapy were included in the 3+3 dose escalation phase. Entinostat was administered orally at a dose of 3 mg once a week (qW) or at a dose of 5 mg qW in combination with a fixed dose of pembrolizumab at a dose of 200 mg every three weeks (q3W), administered via the intravenous route, in a 21-day cycle was studied for security definitions and MTD or RP2D. Evaluation markers correlated with treatment progress included tumor PD-L1 expression and alteration of MDSC and regulatory T cells in peripheral blood samples.

results

Nine patients were included (six of them administered 3 mg entinostat and 3 administered 5 mg qW) with the pembrolizumab combination as above. No DLT was observed in 8 of 9 patients. One patient treated with 3 mg of entinostat developed an increase in alkaline phosphatase and grade 3 bilirubin during cycle 2, which was considered manifestations of immune-mediated hepatitis. This was successfully and rapidly treated by discontinuation of the study drug and administration of systemic corticosteroids. In 2 of 3 patients with paired peripheral blood samples, a decrease in MDSC from baseline was observed. Stable disease was observed in 3 of 6 patients studied.

Conclusion

Entinostat in combination with pembrolizumab proved to be well tolerated and has immunomodulatory activity, as shown by the decrease in MDSC from baseline when the combination was administered.

While the preferred embodiments of the present invention have been shown and described in the present description, it is obvious to a person skilled in the art that such embodiments of the invention are presented by way of example only. Many variations, changes and substitutions are now apparent to those skilled in the art without deviating from the invention. It is to be understood that various alternatives to the embodiments of the invention described herein may be used in the practice of the invention. It is understood that the following claims define the scope of the invention and such methods and structures within the scope of said claims and their equivalents are encompassed therein.

Claims (12)

1. A method of treating melanoma or non-small cell lung cancer in a patient in need thereof, comprising administering to the patient a combination of therapeutic agents, where: therapeutic agents consist of entinostat and pembrolizumab, the patient has previously received a PD-1 blocking antibody or a PD-L1 blocking antibody, and the patient has progressed on previous therapy with a PD-1 blocking antibody or a PD-L1 blocking antibody, where entinostat is administered orally at a dose of 3 mg or 5 mg once a week, and pembrolizumab is administered intravenously at a dose of 200 mg every three weeks, while the duration of the treatment cycle is 21 days. 2. The method of claim 1, wherein the patient has progressed on prior PD-1 blocking antibody therapy. 3. The method of claim 1, wherein the patient has progressed on prior pembrolizumab therapy. 4. The method of claim 1 wherein the initial doses of the combination are administered simultaneously. 5. The method of claim 1 wherein the cancer is melanoma. 6. The method of claim 5 wherein the melanoma is inoperable or metastatic melanoma. 7. The method of claim 1 wherein the cancer is non-small cell lung cancer. 8. The method of claim 7, wherein the non-small cell lung cancer is selected from adenocarcinoma, squamous cell carcinoma, and large cell carcinoma.