Combination of irs / stat3 dual modulators and Anti-PD-1 / PD-l1 antibodies for treating cancer

Abstract

The present invention relates to the treatment of cancer using a combination therapy comprising a dual modulator of insulin receptor substrate (IRS) and signal transduction and activator of transcription protein 3 (Stat3) in combination with an antibody against programmed cell death 1 (PD-1) protein, an antibody against programmed cell death protein 1 ligand (PD-L1), or a combination thereof. The combination may be used to re-sensitize tumors that may or have developed drug resistance to the anti-PD-1 and / or anti-PD-L1 antibodies by enhancing the response of a tumor to the anti-PD-1 and / or anti-PD-L1 antibodies, transforming a non-responsive tumor into a responsive tumor, and / or blocking tumor progression.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of November 16, 2017, an application number of 201780097712.1, and an invention title of "Combination of IRS / STAT3 Dual Modulators and Anti-PD-1 / PD-L1 Antibodies for the Treatment of Cancer". Technical Field

[0002] The present invention relates to the treatment of cancer using a combination therapy that includes a dual modulator of insulin receptor substrate (IRS) and signal transducer and activator of transcription 3 (Stat3) in combination with an antibody against programmed cell death protein 1 (PD-1) and / or an antibody against programmed cell death protein 1 ligand (PD-L1). The combination can be used to resensitize tumors that may develop or have developed resistance to the anti-PD-1 and / or anti-PD-L1 antibodies by enhancing the tumor's response to the anti-PD-1 antibody, converting non-responsive tumors into responsive tumors, and / or blocking tumor progression. Background Art

[0003] Tyrosine phosphorylation inhibitors are a class of protein tyrosine kinase inhibitors that are designed to mimic tyrosine substrates, ATP, and can allosterically inhibit the enzyme (Levitzki et al., Science (1995), 267:1782-88; Levitzki et al., Biochem. Pharm. (1990), 40:913-920; Levitzki et al., FASEB J. (1992), 6:3275-3282; U.S. Patent Nos. 5,217,999 and 5,773,476; Posner et al., Mol. Pharmacol. (1994), 45:673-683). The pharmacophore of these tyrosine phosphorylation inhibitors, especially those of the benzylidenemalononitrile type, is a hydrophilic catechol ring and a more lipophilic substituted cyano-vinyl moiety. Kinetic studies have shown that certain tyrosine phosphorylation inhibitor compounds are pure competitive inhibitors relative to the tyrosine substrate, while they act as non-competitive inhibitors for the ATP binding site (Yaish et al., Science (1988), 242:933-935; Gazit et al., J. Med. Chem. (1989), 32:2344-2352). However, many tyrosine phosphorylation inhibitors have shown competitive inhibition or mixed competition against both the substrate and the ATP binding site (Posner et al., Mol. Pharmacol. (1994), 45:673-683).

[0004] Among a class of related tyrosine phosphorylation inhibitors, the hydrophilic catechol ring is replaced by a lipophilic dichloro- or dimethoxy-phenyl to produce EGFR kinase inhibitors that are effective in the low micromolar range (Yoneda et al., Cancer Res. (1991), 51:4430-4435). Further administration of these tyrosine phosphorylation inhibitors together with a suboptimal dose of anti-EGFR monoclonal antibody to tumor-bearing nude mice provided a significantly enhanced inhibition of tumor growth.

[0005] WO 2008 / 068751, belonging to certain inventors of the present invention, discloses compounds having enhanced inhibitory properties against the activation and signal transduction of insulin-like growth factor 1 receptor (IGF1R), platelet-derived growth factor receptor (PDGFR), epidermal growth factor receptor (EGFR), and IGF1R-related insulin receptor (IR).

[0006] WO 2009 / 147682, belonging to certain inventors of the present invention, discloses compounds that act as regulators of protein kinase (PK) and receptor kinase (RK) signal transduction. Also disclosed in WO 2009 / 147682 are methods for preparing such compounds, pharmaceutical compositions containing such compounds, and methods of using these compounds and compositions, particularly as chemotherapeutic agents for the prevention and treatment of PK- and RK-related disorders such as metabolic, inflammatory, fibrotic, and cell proliferative disorders, particularly cancer.

[0007] WO 2012 / 117396, belonging to certain inventors of the present invention, describes combinations of the compounds of WO 2008 / 068751 or WO 2009 / 147682 with anti-cancer agents for the treatment of cancer.

[0008] WO 2016 / 125169, belonging to certain inventors of the present invention, describes combinations of the compounds of WO 2008 / 068751 or WO 2009 / 147682 with the following agents for the treatment of cancer: (i) epidermal growth factor inhibitors (EGFR inhibitors) and EGFR antibodies; (ii) inhibitors of mammalian target of rapamycin (mTOR); (iii) mitogen-activated protein kinase (MEK) inhibitors; (iv) mutant B-Raf inhibitors; (v) immunotherapeutic agents; and (vi) chemotherapeutic agents.

[0009] In recent decades, immunotherapy has become an important part of the treatment of certain types of cancer. The goal of cancer immunotherapy is to enable the patient's immune system to specifically recognize and kill cancer cells. Signal transducer and activator of transcription 3 (Stat3) is commonly activated in cancer and is directly involved in the implementation and maintenance of the cancer immunosuppressive microenvironment and plays a central role in tumor immune escape.

[0010] There is an unmet need for combinations that can be used to treat cancer and preferably provide at least an additive therapeutic effect. Combinations of drugs from different classes can be used to prevent or overcome the emergence of drug-resistant tumors. SUMMARY OF THE INVENTION

[0011] The present invention relates to the use of a combination therapy for treating cancer, the combination therapy comprising a dual modulator of insulin receptor substrate (IRS) and signal transducer and activator of transcription 3 (Stat3) in combination with an antibody against programmed cell death 1 (PD-1) protein. The combination can be used to resensitize tumors that may develop or have developed resistance to the anti-PD-1 and / or anti-PD-L1 antibodies by enhancing the tumor response to the anti-PD-1 and / or anti-PD-L1 antibodies, converting non-responsive tumors into responsive tumors, and / or blocking tumor progression.

[0012] The compounds described herein are modulators of insulin receptor substrate 1 (IRS1) and / or insulin receptor substrate 2 (IRS2) signaling. Thus, these compounds are referred to herein as "IRS modulators". In certain embodiments, the compounds are inhibitors of IRS1 and / or IRS2. In other embodiments, the compounds of the present invention are inhibitors of insulin-like growth factor 1 receptor (IGF-1R). Thus, these compounds can be used to inhibit, treat, or prevent disorders associated with IGF-1R and / or IRS1 and / or IRS2 signaling, such as cancer. In certain embodiments, the compounds trigger any one or more of the following in any order: (i) dissociation of IRS1 and / or IRS2 from the cell membrane; (ii) serine phosphorylation of the direct substrates IRS1 and / or IRS2 of IGF-1R; and / or (iii) degradation of IRS1 and / or IRS2, thereby providing a long-lasting effect and enhancing the inhibitory activity of these compounds. In other embodiments, the compounds are also inhibitors of IGF1R-related insulin receptor (IR) or proteins that are affected by, mediated by, or part of the signal transduction pathways mediated by these PTKs.

[0013] The compounds described herein are also regulators of signal transducer and activator of transcription 3 (Stat3). Accordingly, these compounds are also referred to as "Stat3 regulators". In certain embodiments, the compounds result in inhibition of Stat3 phosphorylation in cancer cells. Elevated levels of Stat3 phosphorylation have been detected in a variety of different cancers and drug-resistant cancers, leading to increased cancer survival. Additionally, treatment of cancer with PK inhibitor drugs surprisingly results in induction of Stat3 phosphorylation, as demonstrated herein. Without wishing to be bound by any particular theory or mechanism of action, it is contemplated that use of the compounds of the invention to inhibit Stat3 activity may act synergistically with PK inhibitor drugs that upregulate Stat3 as a side effect, may prevent acquired resistance to such drugs, and may be effective against drug-resistant cancers.

[0014] Due to the dual effects on IRS and Stat3, the compounds are further described herein as "IRS / Stat3 dual regulators".

[0015] Dual regulators of IRS and Stat3 have been found to be useful for sensitizing tumors to immunotherapy with anti-PD-1 antibodies. Stat3 is commonly activated in cancer and is directly involved in the implementation and maintenance of the cancer immunosuppressive microenvironment and plays a central role in tumor immune evasion. Without wishing to be bound by any particular theory or mechanism of action, it is contemplated that use of the compounds of the invention to inhibit Stat3 phosphorylation exposes tumors to the local immune system and sensitizes them to immunotherapy with antibodies against PD-1.

[0016] In accordance with the principles of the present invention, it has now been demonstrated that a compound of formula 4, representative of the compounds of formula (I) described herein, in combination with the anti-PD-1 monoclonal antibody pembrolizumab (Keytruda ) converts non-responsive tumors into responsive tumors and blocks tumor progression in a patient-derived xenograft (PDX) model of immunodeficient mice, in which tumors derived from esophageal cancer biopsy tissue are implanted and peripheral blood mononuclear cells (PBMCs) isolated from the blood of the same patient are injected into the mice on the first day of treatment. Although no response was observed with either alone or compound 4 alone and tumor progression was rapid, mice treated with the combination of and compound 4 showed complete blockade of tumor progression. This confirms the ability of the compounds to overcome cancer resistance to anti-PD-1 antibody therapy.

[0017] Accordingly, in one embodiment, the present invention relates to a pharmaceutical combination comprising a compound represented by the structure of formula (I) or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof.

[0018]

[0019] Wherein

[0020] R 1 、R 2 、R 3 、R 5 and R 6 are each independently selected from H, halogen, haloalkyl, and OR 16 , wherein R 16 is H or C1-C4 alkyl;

[0021] R 4 is H or CN; and

[0022] R 7 is H or C1-C4 alkyl.

[0023] In certain embodiments, the compound is represented by the structure of any one of formulas 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. The structures of these compounds are set forth in the detailed description below. The currently preferred combination comprises the compound of formula 4.

[0024] In another embodiment, the present invention relates to a pharmaceutical combination comprising a compound represented by the structure of formula (I) or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof.

[0025]

[0026] Wherein

[0027] A is H or CN;

[0028] X 1 、X 2 、X 3 and X 4 are each independently selected from H, halogen, C1-C4 alkyl, haloalkyl, and OR 1 , wherein R 1 is H or C1-C4 alkyl; and

[0029] X5 is H or C1-C4 alkyl.

[0030] In certain embodiments, the compound is represented by the structure of any one of Formulas 17, 18, 19, 20, 21, 22, 23, 24 or 25. The structures of these compounds are set forth in the detailed description below. The currently preferred combination comprises the compound of Formula 20.

[0031] In another embodiment, the present invention also relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising the step of contacting the tumor with a compound represented by the structure of Formula (I) or (II) or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) antibody and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody.

[0032] In other embodiments, the present invention also relates to a combination comprising a compound of Formula (I) or (II) and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 antibody.

[0033] The currently preferred compound used in the combination of the present invention is the compound of Formula 4. Thus, in another embodiment, the present invention relates to a pharmaceutical combination comprising a compound represented by the structure of Formula 4 or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody.

[0034] In another embodiment, the present invention also relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising the step of contacting the tumor with a compound represented by the structure of Formula 4 or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody.

[0035] In other embodiments, the present invention also relates to a combination comprising a compound of Formula 4 and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody.

[0036] The structure of the compound of Formula 4 is as follows:

[0037]

[0038] In certain embodiments, the anti-PD-1 antibody used in combination with the above compounds is selected from pembrolizumab nivolumab (Opdivo ), pidilizumab (CT-011, MDV9300, Medivation), AGEN-2034, AMP-224, BCD-100, BGBA-317, BI-754091, CBT-501, CC-90006, cemiplimab, GLS-010, IBI-308, JNJ-3283, JS-001, MEDI-0680, MGA-012, MGD-013, PDR-001, PF-06801591, REGN-2810, SHR-1210, TSR-042, LZM-009, and ABBV-181. Each possibility represents an independent embodiment of the present invention.

[0039] In presently preferred embodiments, the anti-PD-1 antibody is pembrolizumab Thus, the presently preferred combination comprises the compound of formula 4 and pembrolizumab.

[0040] In certain embodiments, the anti-PD-L1 antibody used in combination with the above compounds is selected from durvalumab (Imfinzi TM ), atezolizumab (Tecentriq MPDL3280A), avelumab (Bavencio ), CX-072, BMS-936559, SHR-1316, M-7824, LY-3300054, FAZ-053, KN-035, CA-170, CK-301, CS-1001, HLX-10, MCLA-145, MSB-2311, and MEDI-4736. Each possibility represents an independent embodiment of the present invention.

[0041] In certain embodiments, the tumor is present in a cancer patient who is undergoing or is a candidate for immunotherapy using an anti-PD-1 and / or anti-PD-L1 antibody. In certain embodiments, the tumor in the cancer patient is resistant to treatment with an anti-PD-1 and / or anti-PD-L1 antibody alone. As contemplated herein, the compound of formula (I) or formula (II), particularly compound 4, renders the tumor sensitive again to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody by enhancing the tumor's response to the anti-PD-1 and / or anti-PD-L1 antibody, converting non-responsive tumors into responsive tumors, and / or blocking tumor progression.

[0042] The combination of the present invention is suitable for treating various different types of cancer. Specifically, the combination of the present invention is active against head and neck (H&N) cancer, sarcoma, multiple myeloma, ovarian cancer, breast cancer, kidney cancer, gastric cancer, hematopoietic cancer, lymphoma, leukemia including lymphoblastic leukemia, lung cancer, melanoma, glioblastoma, liver cancer, prostate cancer, pancreatic cancer and colon cancer. Each possibility represents an independent embodiment of the present invention.

[0043] As used herein, the term "combination" or "combination therapy" refers to any form of concurrent or parallel therapy using at least two different therapeutic agents. The term is intended to cover the simultaneous administration of the two therapeutic modalities, i.e., using substantially the same treatment schedule, as well as the overlapping administration of each treatment in a sequential or alternating schedule. Each possibility represents an independent embodiment of the present invention.

[0044] The combination therapy is particularly advantageous because the dose of each agent in the combination therapy can be reduced compared to single therapy using each agent, while still achieving an overall anti-cancer effect. Therefore, reducing the dose of each agent may result in reduced side effects. As demonstrated herein, the combination therapy can reduce the development of resistance to a particular anti-cancer treatment and / or cause regression of tumors after the tumors have acquired resistance.

[0045] The compound of formula (I) or formula (II) (e.g., compound 4) and the anti-PD-1 and / or anti-PD-L1 antibody can be administered simultaneously (in the same or separate dosage forms), or they can be administered sequentially in any order. The administration can also be carried out according to an alternating administration schedule, e.g., a compound of formula (I) or (II), then an anti-PD-1 and / or anti-PD-L1 antibody, then another dose of the compound of formula (I) or (II), then the same or another anti-PD-1 and / or anti-PD-L1 antibody, and so on. The present invention contemplates all administration schedules including simultaneous, sequential and alternating, where each possibility represents an independent embodiment of the present invention.

[0046] The pharmaceutical composition of the present invention can be provided in any form known in the art, for example, in a form suitable for oral administration (e.g., solution, suspension, syrup, emulsion, dispersion, tablet, pill, capsule, bolus, granule and powder), parenteral administration (e.g., intravenous, intramuscular, intra-arterial, transdermal, subcutaneous or intraperitoneal), topical administration (e.g., ointment, gel, cream), administration by inhalation or administration by suppository. Each possibility represents an independent embodiment of the present invention.

[0047] Other embodiments of the present invention and the full scope of applicability will become apparent from the detailed description given hereinafter. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various different changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 . In a patient-derived xenograft (PDX) model of immunodeficient mice implanted with tumors from esophageal cancer biopsy tissues, the combination of Compound 4 and pembrolizumab converted non-responsive tumors into responsive tumors and blocked tumor progression. On the first day of treatment, immune cells from the same patient (dual autologous) were replenished in the mice. The mice were treated with (a) control (vehicle) (◇); (b) (□); (c) Compound 4 (Δ); or (d) + Compound 4 (o). Treatment began when the average tumor size was ~160 mm 3 . Although no response was observed when using alone or Compound 4 alone, and the tumors developed rapidly, the mice treated with the combination of and Compound 4 showed complete blockade of tumor progression. DETAILED DESCRIPTION

[0049] The present invention relates to the use of combination therapy for treating cancer, the combination therapy comprising a dual modulator of insulin receptor substrate (IRS) and signal transducer and activator of transcription 3 (Stat3) in combination with an antibody against programmed cell death 1 (PD-1) protein and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody. By enhancing the response of tumors to the anti-PD-1 and / or anti-PD-L1 antibodies, converting non-responsive tumors into responsive tumors, and / or blocking tumor progression, the combination can be used to resensitize tumors that have developed resistance to the anti-PD-1 and / or anti-PD-L1 antibodies.

[0050] PD-1, PD-L1, and anti-PD-1 and PD-L1 antibodies

[0051] Programmed cell death protein 1, also known as PD-1, is a cell surface receptor that plays an important role in downregulating the immune system and promoting self-tolerance by inhibiting the inflammatory activity of T cells. Binding of the PD-1 ligands PD-L1 and PD-L2 to the PD-1 receptor found on T cells inhibits T-cell proliferation and cytokine production. Upregulation of the PD-1 ligand occurs in some tumors, and signaling through this pathway may contribute to the inhibition of active T-cell immunosurveillance of tumors. Anti-PD-1 antibodies bind to the PD-1 receptor and block its interaction with PD-L1 and PD-L3, releasing the inhibition of immune responses, including anti-tumor immune responses, mediated by the PD-1 pathway.

[0052] In certain embodiments, the anti-PD-1 antibody used in combination with the above compounds is pembrolizumab In other embodiments, the anti-PD-1 antibody used in combination with the above compounds is nivolumab In other embodiments, the anti-PD-1 antibody used in combination with the above compounds is pidilizumab (Medivation).

[0053] Other PD-1 antibodies are selected from AGEN-2034 (Agenus), AMP-224 (Medimmune), BCD-100 (Biocad), BGBA-317 (Beigene), BI-754091 (Boehringer Ingelheim), CBT-501 (Genor Biopharma), CC-90006 (Celgene), cemiplimab (Regeneron Pharmaceuticals), durvalumab + MEDI-0680 (Medimmune), GLS-010 (Harbin Gloria Pharmaceuticals), IBI-308 (Eli Lilly), JNJ-3283 (Johnson & Johnson), JS-001 (Shanghai Junshi Bioscience Co.), MEDI-0680 (Medimmune), MGA-012 (MacroGenics), MGD-013 (Marcogenics), pazopanib hydrochloride + pembrolizumab (Novartis), PDR-001 (Novartis), PF-06801591 (Pfizer), REGN-2810 (Regeneron), SHR-1210 (Jiangsu Hengrui Medicine Co.), TSR-042 (Tesaro Inc.), LZM-009 (Livzon Pharmaceutical Group Inc), and ABBV-181 (AbbVie Inc). Each possibility represents an independent embodiment of the present invention.

[0054] In a presently preferred embodiment, the anti-PD-1 antibody is pembrolizumab

[0055] In other embodiments, the anti-PD-L1 antibodies used in the combinations of the present invention are selected from durvalumab (MedImmune LLC), atezolizumab (Hoffmann-La Roche Ltd, Chugai Pharmaceutical Co Ltd), avelumab (Merck KGaA), CX-072 (CytomX Therapeutics Inc), BMS-936559 (ViiV Healthcare Ltd), SHR-1316 (Jiangsu Hengrui Medicine Co Ltd), M-7824 (Merck KGaA), LY-3300054 (Eli Lilly and Co), FAZ-053 (Novartis AG), KN-035 (AlphaMab Co Ltd), CA-170 (Curis Inc), CK-301 (TG Therapeutics Inc), CS-1001 (CStone Pharmaceuticals Co Ltd), HLX-10 (Shanghai Henlius Biotech Co Ltd), MCLA-145 (Merus NV), MSB-2311 (MabSpace Biosciences (Suzhou) Co Ltd), and MEDI-4736 (Medimmune).

[0056] As contemplated herein, the tumors to be treated by the combinations of the present invention are present in cancer patients who are undergoing or are candidates for undergoing immunotherapy with anti-PD-1 and / or anti-PD-L1 antibodies. In certain embodiments, the tumors in such cancer patients are resistant to treatment with anti-PD-1 and / or anti-PD-L1 antibodies alone. As demonstrated herein, compounds of formula (I) or (II), such as compound 4, render the tumors sensitive again to immunotherapy with the anti-PD-1 and / or anti-PD-L1 antibodies by enhancing the response of the tumors to the anti-PD-1 and / or anti-PD-L1 antibodies, converting non-responsive tumors into responsive tumors, and / or blocking tumor progression. Each possibility represents an independent embodiment of the present invention.

[0057] Insulin receptor substrate (IRS) / signal transducer and activator of transcription 3 (Stat3) dual modulator

[0058] Any compound of the structure of formula (I) or (II) or any individual compound embraced by these formulas can be used in the compositions and methods of the present invention.

[0059] The structure of formula (I) is represented as follows:

[0060]

[0061] wherein

[0062] R 1 、R 2 、R 3 、R 5 and R 6 are each independently selected from H, halogen, haloalkyl, and OR 16 , where R 16 is H or C1-C4 alkyl;

[0063] R 4 is H or CN; and

[0064] R 7 is H or C1-C4 alkyl;

[0065] and its salts, hydrates, and solvates.

[0066] In other embodiments, the compound is a compound of formula (I) wherein at least one of R 1 , R 2 , R 3 , R 5 , and R 6 is halogen. The halogen may be F, Cl, Br, or I, and each possibility represents an independent embodiment of the present invention.

[0067] In other embodiments, the compound is a compound of formula (I) wherein at least one of R 1 , R 2 , R 3 , R 5 , and R 6 is haloalkyl.

[0068] In one embodiment, the compound is a compound of formula (I) wherein R 1 is H. In another embodiment, the compound is a compound of formula (I) wherein R 1 is halogen. In another embodiment, the compound is a compound of formula (I) wherein R 1 is haloalkyl. In a particular embodiment, R 1 is F. In another particular embodiment, R 1 is Cl. In another particular embodiment, R 1 is Br. In another particular embodiment, R 1 is I. In another particular embodiment, R 1 is CF3.

[0069] In another embodiment, the compound is one wherein R2 A compound of formula (I) that is H. In another embodiment, the compound is one in which R 2 is a halogen in the compound of formula (I). In a particular embodiment, R 2 is Br.

[0070] In another embodiment, the compound is a compound of formula (I) in which R3 is H. In another embodiment, the compound is one in which R 3 is a halogen in the compound of formula (I). In a particular embodiment, R 3 is Cl. In another particular embodiment, R 3 is Br. In another particular embodiment, R 3 is I.

[0071] In another embodiment, the compound is a compound of formula (I) in which R 4 is H. In another embodiment, the compound is a compound of formula (I) in which R 4 is CN.

[0072] In another embodiment, the compound is a compound of formula (I) in which R 5 is H.

[0073] In another embodiment, the compound is a compound of formula (I) in which R 6 is H. In another embodiment, the compound is a compound of formula (I) in which R 6 is a halogen. In a particular embodiment, R 6 is Br.

[0074] In another embodiment, the compound is a compound of formula (I) in which R 7 is H. In another embodiment, the compound is a compound of formula (I) in which R 7 is a C1-C4 alkyl. In a particular embodiment, R 7 is CH3. In another particular embodiment, R 7 is CH2CH3.

[0075] In one embodiment, the compound is a compound represented by formula 1:

[0076]

[0077] In another embodiment, the compound is a compound represented by formula 2:

[0078]

[0079] In another embodiment, the compound is a compound represented by Formula 3:

[0080]

[0081] In another embodiment, the compound is a compound represented by Formula 4:

[0082]

[0083] In another embodiment, the compound is a compound represented by Formula 5:

[0084]

[0085] In another embodiment, the compound is a compound represented by Formula 6:

[0086]

[0087] In another embodiment, the compound is a compound represented by Formula 7:

[0088]

[0089] In another embodiment, the compound is a compound represented by Formula 8:

[0090]

[0091] In another embodiment, the compound is a compound represented by Formula 9:

[0092]

[0093] In another embodiment, the compound is a compound represented by Formula 10:

[0094]

[0095] In another embodiment, the compound is a compound represented by Formula 11:

[0096]

[0097] In another embodiment, the compound is a compound represented by Formula 12:

[0098]

[0099] In another embodiment, the compound is a compound represented by Formula 13:

[0100]

[0101] In another embodiment, the compound is a compound represented by Formula 14:

[0102]

[0103] In another embodiment, the compound is a compound represented by Formula 15:

[0104]

[0105] In another embodiment, the compound is a compound represented by Formula 16:

[0106]

[0107] Each possibility represents an independent embodiment of the present invention.

[0108] In another embodiment, the compound is represented by the structure of Formula (II):

[0109]

[0110] wherein

[0111] A is H or CN;

[0112] X 1 、X 2 、X 3 and X 4 are each independently selected from H, halogen, C1-C4 alkyl, haloalkyl, and OR 1 wherein R 1 is H or C1-C4 alkyl; and

[0113] X5 is H or C1-C4 alkyl;

[0114] including its salts, hydrates, solvates, polymorphs, optical isomers, geometric isomers, enantiomers, diastereomers, and mixtures.

[0115] In certain embodiments, the compound is a compound of Formula (II) wherein A is H.

[0116] In other embodiments, the compound is a compound of Formula (II) wherein A is CN.

[0117] In other embodiments, the compound is a compound of Formula (II) wherein at least one of X 1 、X 2 、X 3 and X is halogen. The halogen may be F, Cl, Br, or I, and each possibility represents an independent embodiment of the present invention.

[0118] In other embodiments, the compound is one in which X 1 , X 2 , X 3 and X are each H or a halogen in formula (II), wherein the halogen is preferably Cl, Br or I.

[0119] In other embodiments, the compound is one in which X 2 is H in formula (II).

[0120] In other embodiments, the compound is one in which X 5 is H in formula (II).

[0121] In other embodiments, the compound is one in which X 5 is C1-C4 in formula (II).

[0122] Each possibility represents an independent embodiment of the present invention.

[0123] In one embodiment, the compound is the compound represented by formula 17:

[0124]

[0125] In another embodiment, the compound is the compound represented by formula 18:

[0126]

[0127] In another embodiment, the compound is the compound represented by formula 19:

[0128]

[0129] In another embodiment, the compound is the compound represented by formula 20:

[0130]

[0131] In another embodiment, the compound is the compound represented by formula 21:

[0132]

[0133] In another embodiment, the compound is the compound represented by formula 22:

[0134]

[0135] In another embodiment, the compound is the compound represented by formula 23:

[0136]

[0137] In another embodiment, the compound is a compound represented by Formula 24:

[0138]

[0139] In another embodiment, the compound is a compound represented by Formula 25:

[0140]

[0141] The currently preferred compound of formula (II) is the compound of formula 20.

[0142] In other embodiments, the compound is any one of the derivatives described in the following documents: A) PCT International Patent Application Publication No. WO 2008 / 068751; B) PCT International Patent Application Publication No. WO 2009 / 147682; or C) PCT International Patent Application No. WO 2012 / 090204. The content of each of the above references is hereby incorporated by reference in its entirety as if fully set forth herein.

[0143] It should be understood that all conformational isomers, geometric isomers, stereoisomers, enantiomers, and diastereoisomers of any of the compounds described herein are encompassed and can be used in the combinations and methods described in this application.

[0144] All stereoisomers of the above compounds are contemplated, which may be mixtures or may take a pure or substantially pure form. The compounds may have asymmetric centers at any atom. Thus, the compounds may exist in enantiomeric or diastereomeric form or as mixtures thereof. The present invention contemplates the use of any racemate (i.e., a mixture containing equal amounts of each enantiomer), an enantiomerically enriched mixture (i.e., a mixture enriched in one enantiomer), a pure enantiomer or diastereoisomer, or any mixture thereof. Chiral centers may be designated as R or S or R,S or d, D, l, L or d,l, D,L. Compounds containing amino acid residues include residues of D-amino acids, L-amino acids, or racemic derivatives of amino acids. Compounds containing sugar residues include residues of D-sugars, L-sugars, or racemic derivatives of sugars. Residues of D-sugars occurring in nature are preferred. Additionally, several of the compounds of the present invention contain one or more double bonds. The present invention is intended to independently cover all structural and geometric isomers including cis, trans, E, and Z isomers each time they occur.

[0145] One or more compounds of the invention may exist as salts. The term "salt" encompasses both base and acid addition salts, including but not limited to carboxylates and salts formed with amine nitrogen, and includes salts formed with the organic and inorganic anions and cations discussed below. In addition, the term includes salts formed with basic groups (such as amino groups) and organic or inorganic acids by standard acid-base reactions. Such acids include hydrochloric acid, hydrofluoric acid, trifluoroacetic acid, sulfuric acid, phosphoric acid, acetic acid, succinic acid, citric acid, lactic acid, maleic acid, fumaric acid, palmitic acid, cholic acid, pamoic acid, mucic acid, D-glutamic acid, D-camphoric acid, glutaric acid, phthalic acid, tartaric acid, lauric acid, stearic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, sorbic acid, picric acid, benzoic acid, cinnamic acid, etc. Each possibility represents a separate embodiment of the invention.

[0146] The term "organic or inorganic cation" refers to the counterion for the anion of the salt. Such counterions include but are not limited to alkali and alkaline earth metals (such as lithium, sodium, potassium, barium, aluminum, and calcium), ammonium, and mono-, di-, and tri-alkylamines such as trimethylamine, cyclohexylamine, and organic cations such as diphenylmethylammonium, benzylammonium, 2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium, phenethylbenzylammonium, diphenylmethylethylenediammonium, etc. See, for example, Berge et al., J. Pharm. Sci. (1977), 66:1-19, which is incorporated herein by reference.

[0147] The invention also includes solvates of the compounds of the invention and their salts. "Solvate" means a physical association of a compound of the invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bond formation. In some cases, the solvate can be isolated. "Solvate" encompasses both the solution phase and isolable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, etc. "Hydrate" is a solvate in which the solvent molecule is water.

[0148] The invention also includes polymorphs of the compounds of the invention and their salts. The term "polymorph" refers to a specific crystalline or amorphous state of a substance, which can be characterized by specific physical properties such as X-ray diffraction, IR or Raman spectroscopy, melting point, etc.

[0149] Without wishing to be bound by any particular theory or mechanism of action, it is contemplated that the compounds of the present invention are inhibitors of PK signaling, such as IGF-1R. It has now surprisingly been found that, in addition to being inhibitors of IGF-1R, these compounds also cause dissociation of the IGF-1R substrates IRS1 / 2 from the cell membrane, inhibitory serine phosphorylation and / or degradation of the IRS1 / 2 protein. This activity results in long-lasting inhibition of the IGF-1R and IR pathways, growth inhibition of a broad range of cancer cell types, and potent anti-tumor effects. Accordingly, these compounds are referred to as "IRS modulators". In certain embodiments, the compound of formula I is an inhibitor of insulin receptor or insulin-like growth factor-1 receptor (IGF-1R) signaling, and / or the compound of formula I interacts with a substrate protein in the IGF-1R-mediated pathway, or affects or inhibits the substrate protein. In certain embodiments, the substrate protein is insulin receptor substrate 1 (IRS1), insulin receptor substrate 2 (IRS2), or a combination thereof. In a particular embodiment, the compound of formula I is an IGF-1R kinase inhibitor that causes at least one of dissociation of IRS1 or IRS2 from the cell membrane, phosphorylation of IRS1 or IRS2, and / or degradation of IRS1 or IRS2, in any order.

[0150] The compounds described herein are also modulators of signal transducer and activator of transcription 3 (Stat3). In certain embodiments, the compounds cause inhibition of Stat3 phosphorylation in cancer cells. Increased levels of Stat3 phosphorylation have been detected in various different cancers and drug-resistant cancers, leading to increased cancer survival. Without wishing to be bound by any particular theory or mechanism of action, it is contemplated that inhibition of Stat3 activity may synergize with PK inhibitor drugs that upregulate Stat3 as a side effect, may prevent acquired resistance to such drugs, and may be effective against drug-resistant cancers. In addition, Stat3 is commonly activated in cancer and is directly involved in the implementation and maintenance of the cancer immunosuppressive microenvironment and plays a central role in tumor immune evasion. Without wishing to be limited by any particular theory or mechanism of action, it is contemplated that inhibition of Stat3 phosphorylation exposes tumors to the local immune system and renders them sensitive to the local immune system.

[0151] Chemical definition:

[0152] "Alkyl" refers to any saturated aliphatic hydrocarbon, including straight-chain and branched-chain alkyls. In one embodiment, the alkyl has 1-4 carbons and is referred to herein as C1-C4-alkyl. The alkyl may be unsubstituted or substituted with one or more groups selected from halogen, hydroxy, alkoxycarbonyl, acylamino, alkylacylamino, dialkylacylamino, nitro, amino, alkylamino, dialkylamino, carboxyl, thiol, thioalkyl.

[0153] "Hydroxy" means an OH group. "Alkoxy" means -O-alkyl, where R is alkyl as defined above.

[0154] "Amino" means an NH2 group. Alkylamino means -NHR group, where R is alkyl as defined above. Dialkylamino means -NRR' group, where R and R' are alkyl as defined above.

[0155] "Acylamino" means -C(O)NH2 group. Alkylacylamino means -C(O)NHR group, where R is alkyl as defined above. Dialkylacylamino means -C(O)NRR' group, where R and R' are alkyl as defined above.

[0156] When used herein, the term "halogen" or "halo", alone or as part of another group, means chlorine, bromine, fluorine, and iodine. The term "haloalkyl" means an alkyl in which some or all of the hydrogens are independently replaced by halogen groups, including but not limited to trichloromethyl, tribromomethyl, trifluoromethyl, triiodomethyl, difluoromethyl, chlorodifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, bromomethyl, chloromethyl, fluoromethyl, iodomethyl, etc.

[0157] Also included within the scope of the present invention are prodrugs of the compounds disclosed herein. The term "prodrug" refers to a compound that is rapidly converted in vivo, for example by hydrolysis in the blood, into any compound represented by Formula I. Thus, the term "prodrug" refers to a pharmaceutically acceptable precursor of any compound of the present invention. A prodrug may be inactive when administered to a subject, but is converted into an active compound in vivo. The use of prodrugs is particularly advantageous for facilitating the administration of compounds. In mammalian organisms, prodrug compounds generally provide the benefits of solubility, tissue compatibility, or delayed release.

[0158] Cancer treatment

[0159] In one embodiment, the present invention relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising the step of contacting the tumor with a compound represented by the structure of formula (I) or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) antibody and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody.

[0160] In another embodiment, the present invention relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising contacting the tumor with a compound represented by the structure of formula (II) or a salt or hydrate thereof in combination with an anti-programmed cell death protein 1 (PD-1) antibody and / or an anti-programmed cell death protein 1 ligand (PD-L1) antibody.

[0161] In other embodiments, the present invention also relates to a combination comprising a compound of formula (I) and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody.

[0162] In other embodiments, the present invention also relates to a combination comprising a compound of formula (II) and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody.

[0163] A currently preferred compound used in the combinations of the present invention is the compound of formula 4. Thus, in another embodiment, the present invention also relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising contacting the tumor with a compound represented by the structure of formula 4 or a salt or hydrate thereof in combination with an anti-PD-1 and / or anti-PD-L1 antibody.

[0164] In other embodiments, the present invention also relates to a combination comprising a compound of formula 4 and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody.

[0165] Another currently preferred compound used in the combinations of the present invention is the compound of formula 20. Thus, in another embodiment, the present invention also relates to a method of sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, the method comprising contacting the tumor with a compound represented by the structure of formula 20 or a salt or hydrate thereof in combination with an anti-PD-1 and / or anti-PD-L1 antibody.

[0166] In other embodiments, the present invention also relates to a combination comprising a compound of Formula 20 and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, for sensitizing a tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody.

[0167] In certain embodiments, the tumor is present in a cancer patient who is receiving or is a candidate for receiving immunotherapy.

[0168] A currently preferred combination is pembrolizumab in combination with a compound of Formula 4.

[0169] Other currently preferred combinations are pembrolizumab in combination with a compound of Formula 20.

[0170] As used herein, the term "cancer" refers to a disorder in which a population of cells has become unresponsive, to varying degrees, to the control mechanisms that normally govern cell proliferation and differentiation. Cancer refers to a variety of different types of malignant neoplasms and tumors, including primary tumors and metastases. Non-limiting examples of cancers that can be treated with the combination of the present invention are cancers of the brain, ovary, colorectal, pancreas, head and neck, esophagus, prostate, kidney, bladder, breast, lung, oral cavity, and skin.

[0171] The combination of the present invention is suitable for treating a variety of different types of cancers. Specifically, the combination of the present invention is active against head and neck (H&N) cancer, sarcoma, multiple myeloma, ovarian cancer, breast cancer, kidney cancer, gastric cancer, hematological cancer, lymphoma, leukemia including lymphoblastic leukemia, lung cancer, melanoma, glioblastoma, liver cancer, pancreatic cancer, esophageal cancer, prostate cancer, and colon cancer. Each possibility represents an independent embodiment of the present invention.

[0172] Other examples of cancers are: malignant epithelial tumors, sarcomas, myelomas, leukemias, lymphomas, and mixed tumors. Specific categories of tumors include lymphoproliferative disorders, breast cancer, ovarian cancer, prostate cancer, cervical cancer, endometrial cancer, bone cancer, liver cancer, gastric cancer, colon cancer, pancreatic cancer, thyroid cancer, head and neck cancer, cancers of the central nervous system, cancers of the peripheral nervous system, skin cancer, kidney cancer, and metastases of all of the above cancers. Specific types of tumors include hepatocellular carcinoma, liver cancer, hepatoblastoma, rhabdomyosarcoma, esophageal cancer, thyroid cancer, ganglioneuroblastoma, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, infiltrating ductal carcinoma, papillary adenocarcinoma, melanoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma (well-differentiated, moderately-differentiated, poorly-differentiated, or undifferentiated), renal cell carcinoma, adrenocarcinoma, adrenocortical adenocarcinoma, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, nephroblastoma, testicular tumors, lung cancer including small cell, non-small cell, and large cell lung cancer, bladder cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, retinoblastoma, neuroblastoma, colon cancer, rectal cancer, hematopoietic system malignancies including all types of leukemia and lymphoma, including acute myeloid leukemia, acute myelocytic leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, mast cell leukemia, multiple myeloma, myeloid lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, and liver cancer. Each possibility represents an independent embodiment of the present invention.

[0173] In the context of the present invention, the term "treatment of cancer" includes at least one of the following: reducing the growth rate of cancer (i.e., the cancer is still growing, but at a slower rate); stopping the growth of a cancerous growth, i.e., arrest of tumor growth; and, preferably, reduction or decrease in the size of a tumor. The term also includes reducing the number of metastatic tumors, reducing the number of new metastatic tumors formed, slowing the progression of cancer from one stage to another, and reducing cancer-induced angiogenesis. In the most preferred case, the tumor is completely eliminated. Also included in this term is prolonging the survival period of a subject undergoing treatment, prolonging the time to disease progression, tumor regression, etc. It should be understood that the term "treating cancer" also refers to inhibiting the proliferation of malignant (cancer) cells including tumor formation, primary tumors, tumor progression or tumor metastasis. The term "inhibiting proliferation" with respect to cancer cells can also refer to at least one of the following reductions: the number of cells compared to a control (due to cell death, which may be necrosis, apoptosis or any other type of cell death or a combination thereof); a decrease in the cell growth rate, i.e., the total number of cells may increase, but at a lower level or rate compared to the increase in the control; a decrease in the invasiveness of the cells compared to a control group (e.g., as determined by a soft agar assay), even if their total number has not changed; progression from a less differentiated cell type to a more differentiated cell type; a slowdown in tumor transformation; or, a slowdown in the progression of cancer cells from one stage to the next.

[0174] As used herein, the term "administering" means bringing into contact with the combination of the present invention. Administration can be effected to a cell or tissue culture or to a living organism such as a human. In one embodiment, the present invention encompasses administering the combination of the present invention to a human subject.

[0175] A "therapeutic" treatment is the administration of treatment to a subject showing signs of a disease, with the intention of reducing or eliminating those signs. A "therapeutically effective amount" is an amount of a compound or composition sufficient to provide a beneficial effect to the subject to which the compound or composition is administered.

[0176] As used herein, the term "after cessation of treatment" means after the cessation of treatment with a selected drug. For example, according to certain embodiments of the present invention, the length of time required for the IRS / Stat3 dual modulator (e.g., a compound of formula (I) or (II)) to be administered (sequentially or simultaneously) with any of the combination therapies described herein. The treatment (and all compounds) is then stopped, and a period of time is required for tumor monitoring. As contemplated herein, the IRS / Stat3 dual modulators of the present invention are capable of preventing or delaying tumor recurrence to a greater extent after cessation of treatment with any of the combinations described herein compared to administering any of these drugs alone.

[0177] The term “treating a tumor that has developed drug resistance” or “preventing acquired drug resistance of a tumor” for certain anti-cancer drugs means any one or more of the following: (i) the tumor acquires or develops resistance to the anti-cancer drug as a result of the treatment; (ii) the tumor acquires or develops resistance as a result of treatment with other anti-cancer drugs; or (iii) the tumor has intrinsic resistance to the anti-cancer drug.

[0178] From the perspective of differential toxicity associated with two separate treatments, the combination therapy can provide a therapeutic advantage. For example, treatment with one compound can cause specific toxicities not seen when using the other compound, and vice versa. Thus, this differential toxicity can allow each treatment to be administered at a dose at which the toxicity is absent or minimal, such that together the combination therapy provides a therapeutic dose while avoiding the toxicity of each component of the combination agent. Additionally, when the therapeutic effect achieved as a result of the combination treatment is enhanced or is synergistic, i.e., significantly better than the additive therapeutic effect, the dose of each of the agents can even be further reduced, thereby reducing the associated toxicity to an even greater extent.

[0179] The terms “synergistic,” “cooperative,” and “supra-additive” and their grammatical variations are used interchangeably herein. The interaction between an IRS / Stat3 dual modulator and an anti-PD-1 and / or anti-PD-L1 antibody is considered to be synergistic, cooperative, or supra-additive when the effect observed (e.g., cytotoxicity) in the presence of the drugs together is higher than the sum of the individual effects of each drug administered separately. In one embodiment, the observed combined effect of the drugs is significantly higher than the sum of the individual effects. The term “significant” means that p < 0.05 is observed. A non-limiting way to calculate the efficacy of the combination treatment includes using the Bliss additivity model (Cardone et al., Science (1998), 282: 1318–1321), which uses the following formula: Ebliss = EA + EB - EA × EB, where EA and EB are the inhibition fractions obtained with drug A alone and drug B alone at a specific concentration. When the experimentally measured inhibition fraction equals Ebliss, the combination provides an additive therapeutic effect. When the experimentally measured inhibition fraction is greater than Ebliss, the combination provides a synergistic therapeutic effect.

[0180] Pharmaceutical composition

[0181] Although the components of the combination of the present invention can be administered separately, it is contemplated that the components are administered in a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier or excipient. Each of the components can be administered in a separate pharmaceutical composition, or the combination can be administered in one pharmaceutical composition.

[0182] The pharmaceutical compositions of the present invention can be formulated for administration by a variety of different routes, including oral, rectal, transdermal, parenteral (subcutaneous, intraperitoneal, intravenous, intraarterial, transdermal and intramuscular), topical, intranasal or by suppository. Each possibility represents an independent embodiment of the present invention. These compositions are prepared in a manner well known in the pharmaceutical art and contain at least one compound of the present invention as described above as the active ingredient and a pharmaceutically acceptable excipient or carrier. The term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia and can be used in animals and more particularly in humans.

[0183] During the preparation of the pharmaceutical compositions according to the present invention, the active ingredient is generally admixed with a carrier or excipient which can be a solid, semi-solid or liquid material. The compositions can take the form of tablets, pills, capsules, boluses, granules, powders, lozenges, sachets, cachets, elixirs, suspensions, dispersions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders. Each possibility represents an independent embodiment of the present invention.

[0184] The carrier can be any conventionally used carrier and is limited only by chemical-physical considerations such as solubility and lack of reactivity with the compounds of the present invention and the route of administration. The choice of carrier is determined by the specific method for administering the pharmaceutical composition. Some examples of suitable carriers include lactose, glucose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water and methylcellulose. Each possibility represents an independent embodiment of the present invention. The formulations can additionally include lubricants such as talc, magnesium stearate and mineral oil, wetting agents, surfactants, emulsifying agents and suspending agents, preservatives such as methyl and propyl hydroxybenzoates, sweetening agents, flavoring agents, coloring agents, buffering agents (such as acetates, citrates or phosphates), disintegrating agents, humectants, antibacterial agents, antioxidants (such as ascorbic acid or sodium bisulfite), chelating agents (such as ethylenediaminetetraacetic acid), and agents for regulating osmotic pressure such as sodium chloride. Other pharmaceutical carriers can be sterile liquids such as water and oils, including oils of petroleum, animal, vegetable or synthetic origin such as peanut oil, soybean oil, mineral oil, sesame oil, etc., polyethylene glycol, glycerol, propylene glycol or other synthetic solvents. When the pharmaceutical composition is administered intravenously, water is the preferred carrier. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Each possibility represents an independent embodiment of the present invention.

[0185] For the preparation of solid compositions such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid pre - formulation composition which is a homogeneous mixture containing the compound of the present invention. When referring to these pre - formulation compositions as homogeneous, it means that the active ingredient is uniformly dispersed throughout the composition such that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. Then this solid pre - formulation is subdivided into unit dosage forms of the above - mentioned types, which contain, for example, from about 0.1 mg to about 2000 mg, from about 0.1 mg to about 500 mg, from about 1 mg to about 100 mg, from about 100 mg to about 250 mg, etc. of the active ingredient of the present invention.

[0186] Any method can be used to prepare the pharmaceutical composition. Solid dosage forms can be prepared by wet granulation, dry granulation, direct compression, etc. The solid dosage forms of the present invention can be coated or otherwise compounded to provide dosage forms having the advantage of extended action. For example, the tablet or pill can contain an inner dose and an outer dose component, the latter taking the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be released in a delayed manner. A variety of different materials can be used for such enteric layers or coatings, such materials including a large number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate. Each possibility represents an independent embodiment of the present invention.

[0187] Liquid forms for oral or parenteral administration in which the compositions of the present invention can be incorporated include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions using edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical media. Each possibility represents an independent embodiment of the present invention.

[0188] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In one embodiment, the composition is administered via the oral or nasal respiratory route to achieve local or systemic effects. Compositions in pharmaceutically acceptable solvents can be atomized using an inert gas. The atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be attached to a face mask or an intermittent positive - pressure breathing machine. Solution, suspension, or powder compositions can be administered via the oral or nasal route from a device which delivers the formulation in a suitable manner.

[0189] Another formulation suitable for the compositions and methods of the present invention utilizes a transdermal delivery device (“patch”). Such transdermal patches can be used to provide continuous or discontinuous infusion of the compounds of the present invention in a controlled amount. The construction and use of transdermal patches for pharmaceutical delivery are well known in the art.

[0190] In yet another embodiment, the composition is formulated for topical administration, such as as an ointment, gel, drops, or cream. For topical administration to the body surface using, for example, creams, gels, drops, ointments, etc., the compounds of the present invention can be prepared and administered in a physiologically acceptable diluent with or without a pharmaceutical carrier. The present invention can be used topically or transdermally to treat cancer such as melanoma. Adjuvants for topical or gel bases can include, for example, sodium carboxymethylcellulose, polyacrylates, polyethylene oxide-polypropylene oxide block polymers, polyethylene glycols, and cetostearyl alcohol. Each possibility represents a separate embodiment of the present invention.

[0191] Optional formulations include nasal sprays, liposomal formulations, sustained release formulations, pumps for delivering drugs into the body (including mechanical or osmotic pumps), controlled release formulations, etc., as known in the art.

[0192] The composition is preferably formulated as a unit dosage form. The term “unit dosage form” refers to physically discrete units suitable as unit doses for human subjects and other mammals, each unit containing a predetermined quantity of the active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient.

[0193] In preparing the formulation, it may be necessary to comminute the active ingredient to provide a suitable particle size before combining it with the other ingredients. If the active compound is substantially insoluble, it is generally comminuted to a particle size of less than 200 mesh. If the active ingredient is substantially water-soluble, the particle size is usually adjusted by comminution to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.

[0194] It may be necessary to administer the pharmaceutical composition of the present invention topically to the area in need of treatment; this can be achieved by, for example but not limited to, topical infusion during surgery, infusion into the liver by administration to a blood vessel with or without surgery, topical application, for example in combination with a wound dressing after surgery, by injection, e.g., through a catheter, using a suppository, or using an implant, which is a porous, non-porous, or gelatinous material. According to certain embodiments, administration can be by direct injection at the site of the tumor or neoplastic or pre-neoplastic tissue, e.g., by syringe.

[0195] The compounds can also be administered by any convenient route, such as by infusion or bolus injection, by absorption through epithelial layers (such as oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other therapeutically active agents. The administration can be local, or it can be systemic. In addition, it may be necessary to introduce the pharmaceutical composition of the present invention into the central nervous system by any suitable route, including intraventricular injection and intrathecal injection; intraventricular injection can be facilitated, for example, by an intraventricular catheter attached to a reservoir. Pulmonary administration can also be utilized, for example, by using an inhaler or nebulizer and a formulation with a nebulizing agent.

[0196] The compounds of the present invention can be delivered in immediate release or controlled release systems. In one embodiment, an infusion pump can be used to deliver the compounds of the present invention, such as an infusion pump for delivering chemotherapy to a specific organ or tumor (see Buchwald et al., 1980, Surgery 88:507; Saudek et al., 1989, N. Engl. J. Med. 321:574). In one embodiment, the compounds of the present invention are administered in combination with a biodegradable, biocompatible polymeric implant that releases the compound at a controlled rate over a selected period of time at a selected site. Examples of polymeric materials include, but are not limited to, polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene-vinyl acetate and their copolymers and blends. In yet another embodiment, a controlled release system can be placed near the therapeutic target, so that only a fraction of the systemic dose is required.

[0197] In addition, the pharmaceutical composition can sometimes be formulated for parenteral administration (subcutaneous, intravenous, intra-arterial, transdermal, intraperitoneal or intramuscular injection), and can include aqueous and non-aqueous isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostatic agents and solutes that render the preparation isotonic with the blood of the target recipient, as well as aqueous and non-aqueous sterile suspensions, which contain suspending agents, solubilizing agents, thickening agents, stabilizing agents and preservatives. Oils such as petroleum, animal, vegetable or synthetic oils and soaps such as fatty acid alkali metal, ammonium and triethanolamine salts and suitable detergents can also be used for parenteral administration. The above preparations can also be used for direct intratumoral injection. In addition, to minimize or eliminate irritation at the injection site, the composition can contain one or more nonionic surfactants. Suitable surfactants include polyoxyethylene sorbitan fatty acid esters such as sorbitan monooleate, and high molecular weight adducts of ethylene oxide with hydrophobic bases formed by the condensation of propylene oxide with propylene glycol.

[0198] The parenteral preparation may be present in single-dose or multi-dose sealed containers such as ampoules and vials and may be stored under lyophilized (freeze-dried) conditions, with a sterile liquid carrier such as water for injection added just prior to use. Interim injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the types previously described and known in the art. Each possibility represents a separate embodiment of the invention.

[0199] Optionally, the combinations of the invention can be used in hemodialysis such as leukapheresis and other related methods, e.g., by drawing blood from a patient by various different methods such as dialysis through columns / hollow fiber membranes, cartridges, etc., treating it ex vivo with an IRS / Stat3 dual modulator and / or other anti-cancer agents, and returning it to the patient after treatment. Such treatment methods are well-known and described in the art. See, e.g., Kolho et al. (J. Med. Virol. 1993, 40(4): 318-21); Ting et al. (Transplantation, 1978, 25(1): 31-3); the contents of which are incorporated herein by reference in their entirety.

[0200] Dosage and administration schedule

[0201] The treatment using the IRS / Stat3 dual modulator and the anti-PD-1 and / or anti-PD-L1 antibodies can occur in any order, simultaneously, or in combination. For example, the administration of the IRS / Stat3 dual modulator can occur before, after, or simultaneously with the administration of the anti-PD-1 and / or anti-PD-L1 antibodies. For example, a total treatment duration can be determined for the IRS / Stat3 dual modulator. The anti-PD-1 and / or anti-PD-L1 antibodies can be administered before the start of the treatment using the IRS / Stat3 dual modulator or after the treatment using the IRS / Stat3 dual modulator. In addition, the anti-PD-1 and / or anti-PD-L1 antibodies can be administered during the administration of the IRS / Stat3 dual modulator, but need not occur throughout the entire treatment duration. In another embodiment, the treatment regimen includes pre-treatment with the anti-PD-1 and / or anti-PD-L1 antibodies or the IRS / Stat3 dual modulator, followed by the addition of another agent or agents. Alternating dosing sequences are also contemplated. Alternating dosing includes administering the IRS / Stat3 dual modulator and the other anti-PD-1 and / or anti-PD-L1 antibodies in an alternating order, e.g., IRS / Stat3 dual modulator, then anti-PD-1 and / or anti-PD-L1 antibodies, then IRS / Stat3 dual modulator, and so on.

[0202] The amount of the compound effective in the treatment of a particular disorder or condition including cancer will depend on the nature of the disorder or condition and can be determined by standard clinical techniques. In addition, in vitro assays can optionally be used to assist in identifying the optimal dosage range. The exact dosage to be used in the formulation also depends on the route of administration and the progression of the disease or disorder and should be decided according to the judgment of the practitioner and the circumstances of each patient. Preferred dosages are in the range of 0.01 - 1000 mg / kg body weight, 0.1 mg / kg to 100 mg / kg, 1 mg / kg to 100 mg / kg, 10 mg / kg to 75 mg / kg, 0.1 - 1 mg / kg, etc. The (non-limiting) amounts of the IRS / Stat3 dual modulator and the anti-PD-1 and / or anti-PD-L1 antibody include 0.1 mg / kg, 0.2 mg / kg, 0.5 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 50 mg / kg, 60 mg / kg, 75 mg / kg and 100 mg / kg. Alternatively, the amount administered can be measured and expressed as the molar concentration of the compound administered. By way of illustration and not limitation, the IRS / Stat3 dual modulator (e.g., a compound of any one of formulas I, II, III, IV) can be administered in the range of 0.1 - 10 mM, such as 0.1, 0.25, 0.5, 1 and 2 mM. Alternatively, the amount administered can be measured and expressed as mg / ml, μg / ml or ng / ml. By way of illustration and not limitation, the anti-PD-1 and / or anti-PD-L1 antibody can be administered in amounts of 1 ng / ml to 100 mg / ml such as 1 - 1000 ng / ml, 1 - 100 ng / ml, 1 - 1000 μg / ml, 1 - 100 μg / ml, 1 - 1000 mg / ml, 1 - 100 mg / ml, etc. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test bioassays or systems. When a synergistic effect is observed, the total dose of each component can be reduced, so that the side effects experienced by the subject can be significantly reduced while still achieving sufficient anti-cancer effects.

[0203] In one embodiment, the combination therapy reduces the amount of each of its components by 2-fold, i.e., each component is administered at half the dose compared to monotherapy and still achieves the same or similar therapeutic effect. In another embodiment, the combination therapy reduces the amount of each of its components by 5, 10, 20, 50 or 100-fold.

[0204] The dosing schedule depends on several factors such as the cancer to be treated, its severity and progression, the patient population, age, weight, etc. For example, the compositions of the present invention can be administered once a day, twice a day, three times a day, once a week or once a month. Additionally, the administration can be continuous, i.e., daily, or intermittent. As used herein, the term "intermittent" or "intermittently" means stopping and starting at regular or irregular intervals. For example, intermittent administration can be 1 to 6 days per week, or it may mean cyclic administration (e.g., daily administration for 2 to 8 consecutive weeks followed by a rest period of up to one week without dosing), or it may mean alternate-day administration. The different components of the combination can follow different dosing schedules independently of each other.

[0205] To more fully illustrate certain embodiments of the present invention, the following examples are presented. However, they should in no way be construed as limiting the broad scope of the invention. Those skilled in the art can readily devise many variations and modifications of the principles disclosed herein without departing from the scope of the invention.

[0206] Experimental details section

[0207] Example 1. Compound 4 in combination with pembrolizumab The combination converts non-responsive tumors into responsive tumors and blocks tumor progression

[0208] Experimental system :

[0209] Patient-derived xenografts (PDX) of esophageal-gastric junction adenocarcinoma biopsy tissues were subcutaneously (SC) implanted into NodScid mice. On the first day of treatment, peripheral blood mononuclear cells (PBMC) from the same patient were intravenously injected into the mice (double autologous).

[0210] ■ Fresh human primary esophageal-gastric junction adenocarcinoma biopsy tissues were subcutaneously (SC) implanted into NSG mice (P0).

[0211] ■ Subcutaneous implantation of human primary tumor biopsy samples in NodScid mice

[0212] Tumors were extracted from the mice (P3), measured, cut into 1-2 mm pieces and transferred to a gentleMACS tube containing sterile saline. The tumor volume was adjusted to 1.5 mm 3 tumor volume / 100 ul saline. The samples were dissociated using a gentleMACS Octo dissociator. The dissociated tumor tissue was collected using an 18G syringe and injected subcutaneously (approximately 1.5 mm per mouse 3Tumor volume / 100ul saline) was administered to male NOD.CB17-Prkdcscid / J (NodScid) mice. The animals were observed daily and monitored for any discomfort and immobility.

[0213] ■ Isolation of human PBMC

[0214] Human PBMC were isolated from heparinized venous blood samples of the same esophageal adenocarcinoma patients by density gradient centrifugation using Ficol Histopaque (Sigma). Briefly, the heparinized blood was gently layered on LSM medium (e.g., 10 ml blood on 10 ml ficol) at a ratio of 2:1 and centrifuged at 800 g for 10 minutes at RT (centrifuge settings: horizontal rotor, no brake). The white layer representing PBMC was gently aspirated and aseptically transferred to a sterile centrifuge tube. The cell suspension was then washed twice by adding 10 ml PBS and centrifuging at 600 g for 10 minutes to dilute possible Ficol residues. The cell pellet was resuspended in 0.5 ml PBS and cell counting was performed using trypan blue and a hemocytometer. The total cell number and viable cell number were evaluated. The isolated cells were used for cryopreservation in a sterile filtered 10% DMSO / FBS solution, placed in a pre-cooled 4°C Mr. Frosty overnight, and then transferred to -80°C. On the day of injection, the cells were thawed, washed twice by adding PBS and centrifuging (600 g for 10 minutes, RT) to dilute possible DMSO residues, and viable cells were counted.

[0215] ■ More than 90% of the injected mice developed tumors, and treatment was initiated on day 7 when the tumors reached an average size of approximately 160 mm 3 (day 0).

[0216] ■ Treatment:

[0217] On day 0 and 2 hr after the following treatments, PBMC (2.1M viable cells / mouse) were injected into 12 mice:

[0218] 1. Control (20% HPbCD) 50 μl, IV, in 3 mice on days 0, 5, and 11

[0219] 2. Compound 4 70 mg / kg, IV, in 3 mice on days 0, 5, and 11

[0220] 3. Keytruda 6 mg / kg, IP, in 3 mice on days 0, 5, and 11

[0221] 4. Keytruda 6 mg / kg IP + Compound 4 70 mg / kg, IV, in 3 mice on days 0, 5, and 11

[0222] *Pembrolizumab was administered 4 hours after Compound 4

[0223] For each of Treatment Groups 1 - 4, all treatments were initiated simultaneously.

[0224] ■ The length (l) and width (w) of the tumor were measured 4 - 5 times per week, and the tumor volume was calculated as follows: v = lw 2 / 2. The figure represents the mean tumor volume and standard error. Mice were examined 5 times per week and weighed at least twice per week.

[0225] ■ Mice were sacrificed on Day 18 and tumors were harvested for analysis.

[0226] ■ Parallel experiments of esophageal - gastric junction adenocarcinoma PDX were conducted without the patient's PBMC, and the tumor progression rate in the control group was similar to that in the control group in the presence of PBMC.

[0227] Results

[0228] The study was conducted in a patient - derived xenograft (PDX) model of immunodeficient mice implanted with tumors derived from esophageal cancer biopsy tissues, and the mice were supplemented with immune cells from the same patient (dual - autologous). As Figure 1 shown, although no response was observed when using either alone or when using Compound 4 alone, and tumor progression was rapid, mice treated with the and Compound 4 combination showed complete blockade of tumor progression (tumor growth inhibition = 98%, p - value = 0.002). Thus, the combination of Compound 4 with converted non - responsive tumors into responsive tumors and blocked tumor progression in a pre - clinical immuno - oncology model.

[0229] Although certain embodiments of the present invention have been illustrated and described, it is apparent that the present invention is not limited to the embodiments described herein. Numerous modifications, alterations, variations, substitutions, and equivalent forms will be apparent to those skilled in the art without departing from the spirit and scope of the invention as described by the claims.

Claims

1. Use of a compound represented by the structure of formula (4), or a salt or hydrate thereof, in combination with an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, in the manufacture of a medicament for sensitizing a tumor to immunotherapy using an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof.

2. The use according to claim 1, wherein the anti-PD-1 antibody is pembrolizumab (Keytruda) or nivolumab (Opdivo).

3. The use according to any one of claims 1 to 2, wherein the tumor is resistant to treatment with the anti-PD-1 and / or anti-PD-L1 antibody alone, and wherein the compound of formula (4) resensitizes the tumor to immunotherapy using the anti-PD-1 and / or anti-PD-L1 antibody by enhancing the response of the tumor to the anti-PD-1 and / or anti-PD-L1 antibody, converting a non-responsive tumor into a responsive tumor, and / or blocking tumor progression.

4. The use according to any one of claims 1 to 3, wherein the tumor is present in a cancer patient who is receiving or is a candidate for receiving immunotherapy with an anti-PD-1 and / or anti-PD-L1 antibody.

5. The use according to claim 4, wherein the cancer is selected from head and neck (H&N) cancer, esophageal cancer, sarcoma, multiple myeloma, ovarian cancer, breast cancer, kidney cancer, gastric cancer, hematological cancer, lymphoma, leukemia including lymphocytic leukemia, lung cancer, melanoma, glioblastoma, liver cancer, prostate cancer, pancreatic cancer, and colon cancer.

6. The use according to any one of claims 1 to 5, wherein the compound of formula (4) and the anti-PD-1 and / or anti-PD-L1 antibody are administered in the same pharmaceutical composition; or wherein the compound of formula (4) and the anti-PD-1 and / or anti-PD-L1 antibody are in separate pharmaceutical compositions and are administered simultaneously or in any order.

7. The use according to any one of claims 1 to 6, wherein the compound of formula (4) is an inhibitor of the insulin receptor or insulin-like growth factor-1 receptor (IGF-1R), or wherein the compound of formula (4) directly or indirectly interacts with a substrate protein in the IGF-1R-mediated pathway, or affects or inhibits the substrate protein; wherein the substrate protein is insulin receptor substrate 1 (IRS1), insulin receptor substrate 2 (IRS2), or a combination thereof; or wherein the compound causes any one or more of the following in any order: (i) dissociation of IRS1 or IRS2 from the cell membrane; (ii) phosphorylation of IRS1 or IRS2; or (iii) degradation of IRS1 or IRS2; or wherein the compound of formula (4) causes inhibition of Stat3 phosphorylation in cancer cells.

8. A pharmaceutical combination comprising a compound represented by the structure of formula (4) or a salt or hydrate thereof, and an anti-programmed cell death protein 1 (PD-1) antibody, (4) wherein the anti-PD-1 antibody is selected from pembrolizumab (Keytruda) and nivolumab (Opdivo).

9. The combination according to claim 8, wherein the anti-PD-1 antibody is pembrolizumab (Keytruda).

10. The combination according to claim 8, wherein the anti-PD-1 antibody is nivolumab (Opdivo).

11. A pharmaceutical combination comprising a compound represented by the structure of formula (II) or a salt or hydrate thereof, and an anti-programmed cell death protein 1 (PD-1) antibody, an anti-programmed cell death protein 1 ligand (PD-L1) antibody, or a combination thereof, wherein A is H or CN; X 1 、 X 2 、 X 3 and X 4 are each independently selected from H, halogen, C1-C4 alkyl, haloalkyl and OR 1 wherein R 1 is H or C1-C4 alkyl; and X 5 is H or a C1-C4 alkyl group.

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