Combination of small molecule CD-47 inhibitors with other anticancer agents

Abstract

The present invention relates to a composition comprising a CD47-SIRPα blocker and one or more anticancer agents: wherein the CD47-SIRPα blocker is represented by a compound of formula (I). The present invention also relates to a method for treating cancer in a subject by administering a therapeutically effective amount of a CD47-SIRPα blocker represented by formula (I) in combination with one or more anticancer agents.

Description

[0001] Related Applications

[0002] This application claims the benefit of Indian Provisional Application No. 201841042108 filed on November 08, 2018, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention relates to pharmaceutical compositions comprising a small molecule CD-47-SIRPα pathway inhibitor and one or more agents capable of stimulating receptors such as activated Fc receptors (FcR) or other pro-phagocytic receptors. Background Art

[0004] The CD47 / SIRPα axis has been established as a key regulator of myeloid cell activation and serves as an immune checkpoint for macrophage-mediated phagocytosis. Since CD47 is frequently upregulated in several cancers, it contributes to immune evasion and cancer progression. CD47 regulates phagocytosis primarily by interacting with SIRP1α expressed on macrophages. Blockade of SIRP1α / CD47 has been shown to significantly enhance tumor cell phagocytosis and dendritic cell maturation for better antigen presentation, resulting in substantially improved anti-tumor responses in preclinical models of cancer (MP Chao et al. Curr Opin Immunol. 2012(2):225-232). Currently, disruption of the CD47-SIRPα interaction as a therapeutic strategy for cancer is being evaluated by using monoclonal antibodies and engineered receptor decoys targeting CD47 or SIRPα.

[0005] CD47 is expressed on virtually all non-malignant cells, and blocking CD47 or loss of CD47 expression or changes in membrane distribution can be used as a marker for aged or damaged cells, especially on red blood cells (RBCs). Optionally, for those cells where pre-phagocytic signals are also present, blocking SIRPα can also engulf targets that would not normally be engulfed. CD47 is a widely expressed transmembrane glycoprotein with a single Ig-like domain and five transmembrane regions that acts as a cellular ligand for SIRPα, mediating binding through the NH2-terminal V-like domain of SIRPα. SIRPα is primarily expressed on bone marrow cells, including macrophages, granulocytes, myeloid dendritic cells (DCs), mast cells and their precursors, including hematopoietic stem cells.

[0006] CD47 is also constitutively upregulated in many cancers, such as non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), breast cancer, colon cancer, glioblastoma, glioma, ovarian cancer, bladder cancer, and prostate cancer. Tumor cells overexpress CD47, effectively helping tumor cells escape immune surveillance and killing by innate immune cells. However, in most tumor types, blocking the CD47-SIRPα interaction as a single agent may not induce significant phagocytosis and anti-tumor immunity, so it is necessary to combine with other therapeutic agents. In order to maximize the potential of CD-47-SIPRα pathway blockade, the simultaneous involvement of activating receptors such as Fc receptors (FcR) or other pro-phagocytic receptors (collectively referred to as "eat me" signals) may be required.

[0007] The involvement of pro-phagocytic receptors in blocking CD47-SIRPα interactions was demonstrated by the inefficient triggering of phagocytosis by anti-CD47 F(ab) fragments, single-chain variable fragments of CD-47, or SIRPα proteins without an Fc portion. When activated pro-phagocytic receptors are involved, as is evident in the case of blocking anti-CD47 antibodies containing an Fc portion, CD47-SIRPα blockade is able to trigger more efficient phagocytosis. Combining CD47-SIRPα blockers with therapeutic antibodies (containing Fc) targeting tumor antigens stimulates activated Fc receptors (FcRs), resulting in efficient phagocytosis. The Fc portion of therapeutic antibodies targeting tumor antigens also induces antibody-dependent cellular cytotoxicity (ADCC), which also increases the therapeutic efficacy. Therefore, an antibody selected from the group consisting of rituximab, herceptin, trastuzumab, alemtuzumab, bevacizumab, cetuximab and panitumumab, daratumumab can trigger more efficient phagocytosis due to its tumor targeting and ADCC.

[0008] Early approaches to disrupt CD47-SIRPα interactions utilized monoclonal antibodies targeting CD47 or SIRPα and engineered receptor decoys fused to Fc fragments. However, one concern with this approach is that CD47 is highly expressed on both hematopoietic and non-hematopoietic normal cells. Therefore, along with tumor cells, CD47-SIRPα blockers containing Fc portions may also target many normal cells, potentially causing them to be eliminated by macrophages. Blocking the interaction of antibodies with normal cells is considered a major safety issue, leading to anemia, thrombocytopenia, and leukopenia. These agents may also affect macrophage-rich solid tissues such as the liver, lungs, and brain. Therefore, it may be desirable to block the interaction of CD47-SIRPα by agents that do not contain an Fc portion, such as small molecules, peptides, Fab fragments, etc., while activating pro-phagocytic receptors in tumor cells through appropriate combinations to induce efficient phagocytosis of tumor cells.

[0009] In addition to Fc receptors, many other pro-phagocytic receptors have been reported to promote engulfment of tumor cells by triggering phagocytosis in response to CD47-SIRPα blockade. These receptors include SLAMF7, Mac-1, receptors for calreticulin, and possibly receptors yet to be identified. B-cell tumor lines such as Raji and other diffuse large B-cell lymphomas express SLAMF7 and have been implicated in triggering pro-phagocytic signals during CD47-SIRPα blockade.

[0010] Therefore, therapeutic agents known to activate pro-phagocytic receptors are also ideal partners for use in combination with CD47-SIRPα blockade to achieve efficient phagocytosis. These agents include proteasome inhibitors (bortezomib, ixazomib, and carfilzomib), anthracyclines (doxorubicin, epirubicin, daunorubicin, idarubicin, mitoxantrone), oxaliplatin, cyclophosphamide, bleomycin, vorinostat, paclitaxel, 5-fluorouracil, cytarabine, BRAF inhibitors (dabrafenib, vemurafenib), PI3K inhibitors, docetaxel, mitomycin C, C), Sorafenib, Tamoxifen and oncolytic viruses.

[0011] In addition to specific agents known to have effects on 'eat me' signals, other agents, including Abiraterone acetate, Afatinib, Aldesleukin, Aldesleukin, Alemtuzumab, Anastrozole, Axitinib, Belinostat, Bendamustine, Bicalutamide, Blinatumomab, Bosutinib, Brentuximab, Busulfan, Cabazitaxel, Capecitabine, Capecitabine, Carboplatin, Carfilzomib, Carmustine, Ceritinib, Clofarabine, Crizotinib, Dacarbazine, Dactinomycin, Dasatinib, Degarelix, Denileukin, Denosumab, Enzalutamide de), Eribulin, Erlotinib, Everolimus, Exemestane, Exemestane, Fludarabine, Fulvestrant, Gefitinib, Goserelin, Ibritumomab, Imatinib, Ipilimumab, Irinotecan, Ixabepilone, Lapatinib patinib), Lenalidomide, Letrozole, Leucovorin, Leuprolide, Lomustine, Mechlorethamine, Megestrol, Nelarabine, Nilotinib, Nivolumab, Olaparib, Omacetaxine, Palbociclib,Pamidronate, Panitumumab, Panobinostat, Pazopanib, Pegaspargase, Pembrolizumab, Pemetrexed Disodium, Pertuzumab, Plerixafor, Pomalidomide, Ponatinib, Pralatrexate, Procarbazine, Radium 223, Ramucirumab, Regorafenib, rIFNa-2b, Romidepsin, Sunitinib, Temozolomide, Temsirolimus, Thiotepa Thiotepa, Tositumomab, Trametinib, Vinorelbine, Methotrexate, Ibrutinib, Aflibercept, Toremifene, Vinblastine, Vincristine, Idelalisib, Mercaptopurine, and Thalidomide may have an effect on the 'eat me' signaling pathway when combined with CD-47-SIRPα blockers.

[0012] In addition to the above therapeutic agents, other treatment modalities used in cancer therapy also activate pro-phagocytic receptors and can therefore be combined with CD47-SIRPα blockade to achieve efficient phagocytosis. These treatment modalities include hypericin-based photodynamic therapy (Hyp-PDT), radiotherapy, high static pressure, photoporphyrin-based PDT, and rose bengal acetate-based PDT.

[0013] However, there is an unmet need to combine small molecule CD-47-SIRPα pathway inhibitors with agents that can stimulate activating receptors such as Fc receptors (FcRs) or other pro-phagocytic receptors, or with other therapeutic modalities used to activate pro-phagocytic receptors in cancer therapy to maximize the potential of CD-47-SIRPα pathway blockade. Summary of the invention

[0014] The present invention provides a composition comprising a small molecule CD-47-SIRPα pathway inhibitor in combination with an agent that stimulates activated receptors such as Fc receptors (FcR) or other pro-phagocytic receptors, or in combination with other therapeutic modalities for activating pro-phagocytic receptors in cancer therapy to maximize the potential of CD-47-SIRPα pathway blockade.

[0015] In one aspect of the present invention, there is provided herein a composition comprising a CD47-SIRPα blocker and one or more anticancer agents, wherein the CD-47-SIRPα blocker is a small molecule represented by a compound of formula (I):

[0016]

[0017] or a pharmaceutically acceptable salt, amide or ester or a stereoisomer thereof; wherein,

[0018] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring;

[0019] R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl;

[0020] R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0021] In another aspect, the invention relates to a method for treating a subject presenting with CD47 pathway dysregulation, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or amide or ester, or stereoisomer thereof, in combination with one or more anti-cancer agents.

[0022] Another aspect of the present invention provides a method for treating a disease or condition mediated by CD47, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt or amide or ester thereof, or a combination of a stereoisomer thereof and one or more anticancer agents.

[0023] In another aspect of the present invention, provided herein is a combination comprising a CD47-SIRPα blocker and one or more anticancer agents, wherein the small molecule CD-47-SIRPα blocker is represented by a compound of formula (I):

[0024]

[0025] or a pharmaceutically acceptable salt, amide or ester or a stereoisomer thereof; wherein,

[0026] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring;

[0027] R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl;

[0028] R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 : Compound 3 and Compound 6 combined with anti-CD20 antibody enhance the phagocytosis of lymphoma cells

[0031] Figure 2 : Compounds 3 and 6 combined with bortezomib enhance the phagocytosis of multiple myeloma cells

[0032] Figure 3 : Antitumor efficacy of compound 6 alone or in combination with anti-mouse PD-L1 antibody in A20 tumor-bearing mice DETAILED DESCRIPTION

[0033] Each embodiment is provided in a manner to explain the present invention, rather than to limit the present invention. In fact, it is readily understood by those skilled in the art that various modifications and variations may be made to the compounds, compositions and methods described herein without departing from the scope or spirit of the present invention. For example, a feature illustrated or described as part of one embodiment may be applied to another embodiment, thereby obtaining yet another embodiment. Therefore, it is intended that the present invention includes such modifications and variations and their equivalents. Other objects, features and aspects of the present invention are disclosed in or apparent from the following detailed description. It will be understood by those of ordinary skill in the art that this discussion is merely a description of an exemplary embodiment and should not be construed as limiting the broader aspects of the present invention.

[0034] In certain embodiments, the present invention provides a composition comprising a CD47-SIRPα blocker and one or more anticancer agents: wherein the CD47-SIRPα blocker is represented by a compound of formula (I):

[0035]

[0036] or a pharmaceutically acceptable salt, amide or ester or a stereoisomer thereof; wherein,

[0037] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring;

[0038] R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl;

[0039] R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0040] In certain embodiments, the anti-cancer agent is a chemotherapeutic agent or an immunomodulatory agent.

[0041] In certain embodiments, the specific combination partners of interest used with CD47-SIRPα blockers include therapeutic antibodies targeting tumor antigens, which stimulate activation of Fc receptors (FcRs) to produce efficient phagocytosis. In certain embodiments, the specific combination partners of interest include therapeutic antibodies that stimulate Fc receptor-mediated phagocytosis. Therefore, antibodies selected from the group consisting of agents capable of triggering efficient phagocytosis include anti-CD20, such as rituximab, tiuxetan, tositumomab, etc., and their combination is particularly suitable for treating non-Hodgkin's B-cell lymphoma and chronic lymphocytic leukemia (CLL). Combinations with anti-CD22, such as Epratuzumab, etc. are particularly suitable for treating B-cell leukemia and hairy cell leukemia. Combinations with anti-CD52, such as alemtuzumab, etc. are particularly suitable for treating B-cell and T-cell leukemias, including but not limited to chronic lymphocytic leukemia. The combination with anti-CD33, for example gemtuzumab ozogomicin, etc. is particularly suitable for treating myeloid leukemia, such as acute myeloid leukemia. The combination with trastuzumab is particularly suitable for treating breast cancer. The combination with bevacizumab is particularly suitable for treating certain types of brain tumors and certain types of renal cancer, lung cancer, colon cancer, rectal cancer, cervical cancer, ovarian cancer or fallopian tube cancer. The combination with cetuximab is particularly suitable for treating colon cancer and head and neck cancer. The combination with panitumumab is particularly suitable for treating colorectal cancer. The combination with daratumumab is particularly suitable for treating multiple myeloma. Other combinations of interest for treating myeloid leukemia include but are not limited to anti-CD96, anti-CD44 and anti-CD123.

[0042] In certain preferred embodiments, the Fc receptor (FcR) comprises an Fc gamma receptor (FcγR).

[0043] Other therapeutic antibodies of interest in combination with CD47-SIRPα blockers include, but are not limited to, ofatumumab for chronic lymphocytic leukemia, obinutuzumab for follicular lymphoma, alemtuzumab for B-cell chronic lymphocytic leukemia, Ibritumomab tiuxetan for B-cell non-Hodgkin lymphoma, dinutuximab for neuroblastoma, and necitumumab for lung cancer.

[0044] In certain embodiments, the anti-cancer agent is an anti-CD20 antibody, such as rituximab, tazetam, tositumomab.

[0045] In certain embodiments, therapeutic agents known to activate pro-phagocytic receptors are therefore ideal partners for use in combination with CD47-SIRPα blockers to achieve efficient phagocytosis. These agents include proteasome inhibitors (bortezomib, ixazomib and carfilzomib), anthracyclines (doxorubicin, epirubicin, daunorubicin, idarubicin, mitoxantrone) oxaliplatin, cyclophosphamide, bleomycin, vorinostat, paclitaxel, 5-fluorouracil, cytarabine, BRAF inhibitory drugs (dabrafenib, vemurafenib), PI3K inhibitors, docetaxel, mitomycin C, sorafenib and tamoxifen; or combinations thereof.

[0046] In certain embodiments, the anticancer agent is a proteasome inhibitor.

[0047] In certain embodiments, the anticancer agent is bortezomib, ixazomib, or carfilzomib, or an analog or derivative thereof.

[0048] In certain embodiments, in addition to specific agents known to have an effect on the 'eat me' signal, other agents, including abiraterone acetate, afatinib, aldesleukin, aldesleukin, alemtuzumab, anastrozole, axitinib, belinomastat, bendamustine, bicalutamide, blinatumomab, bosutinib, bentuximab, busulfan, cabazitaxel, capecitabine, carboplatin, carfilzomib, carmustine, cerevisiae, tinib, clofarabine, crizotinib, dacarbazine, actinomycin D, dasatinib, degarelix, denileukin, denosumab, enzalutamide, eribulin, erlotinib, everolimus, exemestane, exemestane, fludarabine, fulvestrant, gefitinib, goserelin, ibritumomab tiuxetan, imatinib, ipilimumab, irinotecan, ixabepilone, lapatinib, lenalidomide, letrozole, formyl Folic acid, leuprolide, lomustine, meclofenamic acid, megestrol acetate, nelarabine, nilotinib, nivolumab, olaparib, omacitaxin, palbociclib, pamidronate, panitumumab, panobinostat, pazopanib, pegaspargase, pembrolizumab, pemetrexed disodium, pertuzumab, plerixafor, pomalidomide, ponatinib, pralatrexate, procarbazine, radium 223, ramucirumab , regorafenib, rIFNa-2b, romidepsin, sunitinib, temozolomide, temsirolimus, thiotepa, tositumomab, trametinib, vinorelbine, methotrexate, ibrutinib, aflibercept, toremifene, vinblastine, vincristine, idelalisib, mercaptopurine, and thalidomide may have an effect on the ‘eat me’ signaling pathway when combined with CD-47-SIRPα blockers.

[0049] In other embodiments, in addition to the above therapeutic agents, other therapeutic modalities used in cancer therapy can also activate pro-phagocytic receptors and can therefore be combined with CD47-SIRPα blockers to achieve efficient phagocytosis. These therapeutic modalities include hypericin-based photodynamic therapy (Hyp-PDT), radiotherapy, high static pressure, photoporphyrin-based PDT, and rose bengal acetate-based PDT.

[0050] In certain embodiments, the chemotherapeutic agent is abarelix, aldesleukin, alitretinoin, allopurinol, altretamine, arsenic trioxide, asparaginase, azacitidine, bexarotene, baricitinib, bortezomib, intravenous busulfan, oral busulfan, calusterone, cetuximab, chlorambucil, cisplatin, cladribine, dalteparin sodium, decitabine, diftitox, disulfiram, dexrazoxane, dromostanolone propionate, propionate, eculizumab, estramustine, etoposide phosphate, etoposide, fentanyl citrate, filgrastim, floxuridine, gemcitabine, histrelin acetate, fosfamide, interferon α2a, lapatinib ditosylateditosylate, levamisole, marizomib, meclorethamine, melphalan, mercaptopurine, methotrexate, methoxsalen, mitotane, nandrolone phenpropionate, nofetumomab, oprozomib, pegfilgrastim, pentostatin tin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, ruxolitinib, rucaparib, streptozocin, teniposide, testolactone, thalidomide, thioguanine, topotecan, toremifene, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, niraparib, veliparib, talazoparib, or zoledronate.

[0051] In certain embodiments, the anticancer agent is an immunomodulator. In further embodiments, the immunomodulator is a co-stimulatory or co-inhibitory molecule, such as CTLA-4 (e.g., ipilimumab), 4-1BB (e.g., urelumab and utomilumab), antibodies to PD-1 and PD-L1 (e.g., nivolumab, pembrolizumab, atezolizumab, durvalumab and camrelizumab), cytokine antibodies (IL-10, TGF-β), TIM-3 antibodies, LAG3 antibodies, B7H3 antibodies, B7H4 antibodies, and B7H6 antibodies; or combinations thereof.

[0052] In some embodiments of the invention, two or more CD47-SIRPα blockers represented by compounds of formula (I) are administered. In some embodiments, the CD47-SIRPα blocker is administered more than once.

[0053] In certain embodiments, a CD47-SIRPα blocking agent is an agent that blocks the interaction between CD47 and SIRPα. In certain embodiments, blocking the interaction between CD47 and SIRPα induces macrophages to phagocytose tumor cells expressing CD47.

[0054] In certain embodiments, the present invention provides a composition comprising a small molecule CD47-SIRPα blocker as described in the compound of formula (I) and a proteasome inhibitor. In certain embodiments, the proteasome inhibitor is bortezomib, ixazomib or carfilzomib or any analog or derivative thereof.

[0055] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R1 is hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2-phenyl, or -CH2-imidazolyl.

[0056] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R1 is -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, or -(CH2)4NH2.

[0057] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R1 is -(CH2)2CONH2 or -(CH2)2COOH.

[0058] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-phenyl or -CH2-imidazolyl.

[0059] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R2 is hydrogen, -(CH2)3NHC(=NH)NH2, or -(CH2)2COOH.

[0060] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R2 is hydrogen, -(CH2)2CONH2, or -(CH2)2COOH.

[0061] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R3 represents hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-imidazolyl.

[0062] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R3 represents hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH or -(CH2)4NH2.

[0063] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-phenyl or -CH2-imidazolyl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring;

[0064] R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-phenyl or -CH2-imidazolyl;

[0065] R b is hydrogen; and R3 represents hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-imidazolyl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0066] In certain embodiments, the compositions of the invention comprise a compound of formula (I), which is a compound of formula (IA):

[0067]

[0068] or its pharmaceutically acceptable salts, amides or esters or their stereoisomers; wherein R1, R a and R2 are as defined for the compounds of formula (I).

[0069] In certain embodiments, the compositions of the present invention comprise a compound of formula (IA), wherein R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2-phenyl or -CH2-imidazolyl.

[0070] In certain embodiments, the compositions of the present invention comprise a compound of formula (IA), wherein R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-phenyl or -CH2-imidazolyl.

[0071] In certain embodiments, the compositions of the invention comprise a compound of formula (IA), wherein R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2-phenyl or -CH2-imidazolyl; R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-phenyl or -CH2-imidazolyl.

[0072] In another embodiment, the composition of the present invention comprises a compound of formula (IA): wherein R a R1 represents -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2 or -(CH2)4NH2. R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2COOH, -CH2-phenyl or -CH2-imidazolyl.

[0073] In certain embodiments, the compositions of the present invention comprise a compound of formula (I) selected from the group consisting of,

[0074] or

[0075]

[0076] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0077] In certain embodiments, the compositions of the present invention comprise a compound of formula (I) selected from the group consisting of,

[0078] or

[0079] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0080] In certain embodiments, the compositions of the invention comprise a compound of formula (IA), which may also be written by indicating its absolute stereochemistry, such as

[0081]

[0082] In certain embodiments, the compositions of the invention comprise a compound of formula (I), which is a compound of formula (IB):

[0083]

[0084] or its pharmaceutically acceptable salts, amides or esters or their stereoisomers; wherein R1, R a , R b and R3 are as defined for the compound of formula (I).

[0085] In certain embodiments, the compositions of the present invention comprise compounds wherein R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, or -CH2-phenyl.

[0086] In certain embodiments, the compositions of the present invention comprise compounds wherein R1 is -(CH2)2CONH2 or -(CH2)2COOH.

[0087] In certain embodiments, the compositions of the present invention comprise compounds wherein R3 is hydrogen, -CH2-phenyl, -(CH2)2CONH2, or -(CH2)2COOH.

[0088] In certain embodiments, the compositions of the present invention comprise compounds wherein R3 is hydrogen or -CH2-phenyl.

[0089] In certain embodiments, the compositions of the invention comprise a compound of formula (IB), wherein R b For hydrogen.

[0090] In certain embodiments, in Formula (IB), R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0091] In certain embodiments, the compositions of the present invention comprise a compound of formula (IB), wherein R1 represents -(CH2)2CONH2 or -(CH2)2COOH; R b is hydrogen; and R3 represents hydrogen or -CH2-phenyl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0092] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein the compound is selected from the group consisting of,

[0093] or

[0094] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0095] In certain embodiments, the compositions of the invention comprise a compound of formula (I), which is a compound of formula (IC):

[0096]

[0097] or its pharmaceutically acceptable salts, amides or esters or their stereoisomers; wherein R1, R a , R3 and R b As defined for compounds of formula (I).

[0098] In certain embodiments, the compositions of the present invention comprise compounds wherein R1 is -(CH2)2CONH2, -(CH2)3NHC(=NH)NH2, or -(CH2)4NH2.

[0099] In certain embodiments, the compositions of the present invention comprise compounds wherein R1 is -(CH2)2CONH2 or -(CH2)3NHC(=NH)NH2.

[0100] In certain embodiments, the compositions of the present invention comprise a compound of formula (IC), wherein R a In certain embodiments, in Formula (IC), R a and R1 together with the atoms to which they are attached form a pyrrolidine ring.

[0101] In certain embodiments, the compositions of the present invention comprise compounds wherein R3 is hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2, or -CH2-imidazolyl.

[0102] In certain embodiments, the compositions of the present invention comprise a compound of formula (IC), wherein R b For hydrogen.

[0103] In certain embodiments, in Formula (IC), R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0104] In another embodiment, the composition of the present invention comprises a compound of formula (IC): wherein R a is hydrogen; and R1 represents -(CH2)2CONH2, -(CH2)3NHC(=NH)NH2 or -(CH2)4NH2; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring; and R bis hydrogen; and R3 represents -CH2-phenyl, -(CH2)3NHC(=NH)NH2 or -CH2-imidazolyl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0105] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein the compound is selected from the group consisting of,

[0106] or

[0107] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0108] In certain embodiments, the compositions of the invention comprise a compound of formula (I), which is a compound of formula (ID):

[0109]

[0110] or its pharmaceutically acceptable salts, amides or esters or their stereoisomers; wherein R1, R a , R3 and R b As defined for compounds of formula (I).

[0111] In certain embodiments, the compositions of the present invention comprise compounds wherein R1 is -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, or -CH2CONH2.

[0112] In certain embodiments, the compositions of the present invention comprise compounds wherein R3 is hydrogen, -(CH2)3NHC(=NH)NH2, or -(CH2)4NH2.

[0113] In certain embodiments, the compositions of the present invention comprise a compound of formula (ID), wherein R b For hydrogen.

[0114] In certain embodiments, in Formula (ID), R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0115] In another embodiment, the composition of the present invention comprises a compound of formula (ID): wherein R1 represents -(CH2)3NHC(=NH)NH2, -(CH2)4NH2 or -CH2CONH2; R b is hydrogen; and R3 represents hydrogen, -(CH2)3NHC(=NH)NH2 or -(CH2)4NH2; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0116] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein the compound is selected from the group consisting of,

[0117] or

[0118]

[0119] Or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof. In certain embodiments, the composition of the present invention comprises a compound of formula (I), which is a compound of formula (IE):

[0120]

[0121] or its pharmaceutically acceptable salts or amides or esters or their stereoisomers; wherein R2, R3 and R b As defined for compounds of formula (I).

[0122] In certain embodiments, the compositions of the present invention comprise compounds wherein R2 is hydrogen or -(CH2)3NHC(=NH)NH2.

[0123] In certain embodiments, the compositions of the present invention comprise a compound of formula (IE), wherein R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0124] In another embodiment, the composition of the present invention comprises a compound of formula (IE): wherein R2 represents hydrogen or -(CH2)3NHC(=NH)NH2; R b and R3 together with the atoms to which they are attached form a pyrrolidine ring. In certain embodiments, the compositions of the present invention comprise a compound of formula (I), which is a compound of formula (IF):

[0125]

[0126] or its pharmaceutically acceptable salts or amides or esters or their stereoisomers; wherein R2, R3 and R b As defined for compounds of formula (I).

[0127] In certain embodiments, the compositions of the present invention comprise compounds wherein R2 is hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2, or -(CH2)2COOH.

[0128] In certain embodiments, the compositions of the present invention comprise compounds wherein R3 is -CH2-phenyl, -(CH2)2CONH2, or -(CH2)2COOH.

[0129] In certain embodiments, the compositions of the invention comprise a compound of formula (IF), wherein R b For hydrogen.

[0130] In certain embodiments, in Formula (IF), R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0131] In a particular embodiment, the composition of the present invention comprises a compound of formula (IF): wherein R2 represents hydrogen, -CH2-phenyl, -(CH2)3NHC(=NH)NH2 or -(CH2)2COOH; R b is hydrogen; and R3 represents -CH2-phenyl, -(CH2)2CONH2 or -(CH2)2COOH; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0132] In certain embodiments, the compositions of the present invention comprise a compound of formula (I), wherein the compound is selected from the group consisting of,

[0133] or

[0134]

[0135] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0136] In certain embodiments, the compositions of the present invention comprise a compound, wherein the compound is selected from:

[0137]

[0138]

[0139]

[0140]

[0141] or a pharmaceutically acceptable salt or amide or ester or a stereoisomer thereof.

[0142] In certain embodiments, the present invention relates to a composition comprising a CD47-SIRPα blocker and one or more anti-cancer agents for use as a medicament, wherein the CD47-SIRPα blocker is a compound of formula (I) or a pharmaceutically acceptable salt or amide or ester thereof, or a stereoisomer thereof, as described herein; and the anti-cancer agent is a chemotherapeutic agent or an immunomodulatory agent, as described herein.

[0143] In certain embodiments, the present invention relates to a composition comprising a CD47-SIRPα blocker and one or more anti-cancer agents and a pharmaceutically acceptable carrier, wherein the CD47-SIRPα blocker is a compound of formula (I) or a pharmaceutically acceptable salt or amide or ester thereof, or a stereoisomer thereof, as described herein; and the anti-cancer agent is a chemotherapeutic agent or an immunomodulatory agent, as described herein.

[0144] In certain embodiments, the present invention provides a combination comprising a CD47-SIRPα blocker and one or more anticancer agents: wherein the CD47-SIRPα blocker is represented by a compound of formula (I):

[0145]

[0146] or a pharmaceutically acceptable salt, amide or ester or a stereoisomer thereof; wherein,

[0147] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring;

[0148] R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl;

[0149] R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0150] In certain embodiments, the combinations of the invention comprise a CD47-SIRPα blocker of a compound of formula (I) and a proteasome inhibitor.

[0151] In certain embodiments, the combination of the invention comprises a CD47-SIRPα blocker of a compound of formula (I) and an anti-CD-20 antibody.

[0152] In certain embodiments, the combinations of the invention comprise a CD47-SIRPα blocker of a compound of Formula (IA), (IB), (IC), (ID), (IE) or (IF).

[0153] In certain embodiments, the present invention relates to a method of treating cancer in a subject presenting with CD47 pathway dysregulation, the method comprising administering to the subject a therapeutically effective amount of a CD47-SIRPα blocker in combination with a therapeutically effective amount of one or more anti-cancer agents: wherein the CD47-SIRPα blocker is represented by a compound of formula (I):

[0154]

[0155] or a pharmaceutically acceptable salt, amide or ester thereof; or a stereoisomer thereof;

[0156] in,

[0157] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring; R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl; R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0158] In certain embodiments, a subject presenting a dysregulation of the CD47 pathway is a subject presenting a CD47+ disease cell. In certain embodiments, the CD47+ disease cell is a CD47+ cancer cell.

[0159] In certain embodiments, the invention relates to a method wherein treatment with one or more anticancer agents is prior to, concurrently with, or subsequent to treatment with a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0160] In certain embodiments, the present invention relates to a method for treating a disease or condition mediated by the CD47 pathway in a subject or delaying the progression of the disease or condition, the method comprising administering to a subject in need thereof a therapeutically effective amount of a combination of an agent that blocks the CD47-SIRPα pathway and a therapeutically effective amount of one or more anti-cancer agents: wherein the agent that blocks the CD47-SIRPα pathway is represented by a compound of formula (I):

[0161]

[0162] or a pharmaceutically acceptable salt, amide or ester thereof; or a stereoisomer thereof;

[0163] in,

[0164] R a is hydrogen; and R1 represents hydrogen, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)4NH2, -CH2CONH2, -CH2-aryl or -CH2-heteroaryl; or R a and R1 together with the atoms to which they are attached form a pyrrolidine ring; R2 represents hydrogen, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -CH2-aryl or -CH2-heteroaryl; R b is hydrogen; and R3 represents hydrogen, -CH2-aryl, -(CH2)3NHC(=NH)NH2, -(CH2)2CONH2, -(CH2)2COOH, -(CH2)4NH2 or -CH2-heteroaryl; or R b and R3 together with the atoms to which they are attached form a pyrrolidine ring.

[0165] In certain embodiments, the invention relates to a method, wherein the disease or condition mediated by the CD47-SIRPα pathway is cancer.

[0166] In certain embodiments, the invention relates to a method wherein the disease or condition mediated by the CD47 pathway is atherosclerosis.

[0167] In certain embodiments, the invention relates to a method wherein the disease or disorder mediated by the CD47 pathway is multiple sclerosis.

[0168] In certain embodiments, the present invention provides a pharmaceutical composition comprising a compound as disclosed herein, optionally in admixture with a pharmaceutically acceptable carrier or diluent.

[0169] In certain embodiments, the present invention provides for use of a composition comprising a CD47-SIRPα blocker and one or more anti-cancer agents as described herein in the manufacture of a medicament for treating cancer in a subject exhibiting CD47 pathway dysregulation.

[0170] In certain embodiments, the invention provides a kit comprising a composition as described herein and a package insert comprising instructions for administration of the drug for treating a subject exhibiting a CD47 pathway dysregulation.

[0171] The present invention also provides methods for formulating the disclosed compounds for pharmaceutical administration.

[0172] The compositions and methods of the present invention can be used to treat individuals in need. In certain embodiments, the individual is a mammal, such as a human or non-human mammal. When applied to an animal such as a human, the compound is preferably administered in the form of a pharmaceutical composition, which comprises, for example, the compounds of the present invention and a pharmaceutically acceptable carrier. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive administration routes (i.e., routes of transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of the agent or selective targeting of one or more cells, tissues or organs. The pharmaceutical composition can be in the form of a dosage unit, such as a tablet, a capsule (including a sprinkle capsule and a gelatin capsule), a granule, a lyophilized agent for reconstruction, a powder, a solution, a syrup, a suppository, an injection, etc. The composition can also be present in a transdermal delivery system, such as a skin patch. The composition can also be present in a solution suitable for topical administration, such as an eye drop.

[0173] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that acts, for example, to stabilize, increase solubility, or increase the absorption of a compound. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier (including a physiologically acceptable agent) depends, for example, on the route of administration of the composition. The preparation of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation agent) may also be a liposome or other polymer matrix, into which, for example, a compound of the present invention may be incorporated. Liposomes, such as liposomes comprising phospholipids or other lipids, may be relatively simple, non-toxic, physiologically acceptable, and metabolizable carriers to manufacture and administer.

[0174] The agents in the composition are administered simultaneously, i.e., each agent is administered within about 45 days, 30 days, 15 days, 7 days, 3 days, 2 days, 1 day, or substantially simultaneously with respect to the other agents in the composition. If the administration schedule is such that the serum levels of the two agents reach therapeutic levels, the agents are considered to be used in combination.

[0175] In certain embodiments, for administration, each dose of the combination of CD47-SIRPα blocker and anticancer agent will be in the range of about 0.0001 to 100 mg / kg, more typically 0.01 to 50 mg / kg of the host body weight. For example, the dose can be 1 mg / kg body weight, 10 mg / kg body weight, or 30 mg / kg body weight, or in the range of 1-50 mg / kg. The dose can be adjusted according to the molecular weight of the CD47-SIRPα blocker or anticancer agent, and can be reduced relative to the dose required for monotherapy of any agent in the combination. Exemplary treatment regimens require administration daily, half a week, weekly, once every two weeks, once a month, etc.

[0176] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0177] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical compositions.

[0178] The pharmaceutical composition (preparation) can be administered to a subject by any of a variety of routes of administration, including, for example, oral routes (e.g., syrups, tablets, capsules (including sprinkle capsules and gelatin capsules) in the form of aqueous solutions or non-aqueous solutions or suspensions, boluses, powders, granules, pastes applied to the tongue); absorption routes through the oral mucosa (e.g., sublingual); anal, rectal or vaginal routes (e.g., as vaginal suppositories, creams or foams); parenteral routes (including intramuscular, intravenous, subcutaneous or intrathecal, e.g., as sterile solutions or suspensions); nasal routes; intraperitoneal routes; subcutaneous routes; transdermal routes (e.g., as patches attached to the skin); and topical routes (e.g., as creams, ointments or sprays applied to the skin or as eye drops). The compound can also be formulated for inhalation. In certain embodiments, the compound can be simply dissolved or suspended in sterile water.

[0179] The preparation can be conveniently present in unit dosage form and can be prepared by any method known in the pharmaceutical field. The amount of each active ingredient in the composition that can be combined with a carrier material to produce a single dosage form will vary according to the host being treated, the specific mode of administration. The amount of each active ingredient in the composition that can be combined with a carrier material to produce a single dosage form will generally be the amount of the compound that produces the therapeutic effect. In general, in one hundred percent, this amount will be between about 1% to about 99% of each active ingredient, preferably about 5% to about 70%, and most preferably about 10% to about 30%.

[0180] The method for preparing these preparations or compositions comprises the step of combining the composition of the present invention with a carrier and optionally one or more auxiliary ingredients. In general, the preparations are prepared by uniformly and intimately bringing into association a compound of the present invention with a liquid carrier, or a finely divided solid carrier, or both, and then, if necessary, shaping the product.

[0181] Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilizates, powders, granules, or as a solution or suspension in an aqueous liquid or non-aqueous liquid, or as an oil-in-water or water-in-oil emulsion, or as an elixir or syrup, or as a pastille (using an inert base such as gelatin and glycerin, or sucrose and acacia) and / or as a mouthwash, etc., each containing a predetermined amount of the composition of the invention as a therapeutically active combination. The composition may also be administered as a bolus, electuary or paste.

[0182] To prepare a solid dosage form for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, lozenges, powders, granules, etc.), the composition is mixed with one or more pharmaceutically acceptable carriers (such as sodium citrate or dicalcium phosphate) and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose and / or gum arabic; (3) wetting agents, Such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution delay agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glyceryl monostearate; (8) absorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules using excipients (such as lactose or milk sugar) and high molecular weight polyethylene glycols.

[0183] Tablets can be prepared by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium carboxymethyl starch or cross-linked sodium carboxymethyl cellulose), a surfactant, or a dispersant. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0184] Other solid dosage forms of the tablets and pharmaceutical compositions, such as lozenges, capsules (including sprinkle capsules and gelatin capsules), pills and granules, can be optionally scored or prepared with coatings and shells (such as enteric coatings and other coatings well-known in the field of pharmaceutical preparations). Hydroxypropyl methylcellulose, other polymer matrices, liposomes and / or microspheres in different proportions that provide the desired release profile can also be used to prepare them so that the combined active ingredients therein are slowly or controlled released. They can be sterilized by, for example, filtering through a filter that retains bacteria or by immediately incorporating a sterilizing agent in the form of a sterile solid composition that is soluble in sterile water or some other sterile injection medium before use. These compositions may also optionally contain an opacifier and may also be a composition that releases the active ingredient only or preferentially in a certain part of the gastrointestinal tract in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in a microencapsulated form and appropriately have one or more of the above-mentioned excipients.

[0185] Liquid dosage forms that can be used for oral administration include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as, for example, water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof.

[0186] Besides inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0187] Suspensions, in addition to the active ingredients, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0188] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration may be presented as suppositories which may be prepared by mixing one or more active ingredients of the compositions of the present invention with one or more suitable non-irritating excipients or carriers (including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate) and which are solid at room temperature but liquid at body temperature and will therefore melt in the rectum or vaginal cavity and release the active ingredient.

[0189] Formulations of pharmaceutical compositions for oral administration may be in the form of a mouthwash, oral spray, or oral ointment.

[0190] Alternatively or additionally, the composition can be configured for delivery via a catheter, stent, wire or other intraluminal device. Delivery via such a device may be particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[0191] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0192] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0193] In addition to the active compounds, the ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0194] In addition to the active compounds, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorocarbons and volatile unsubstituted hydrocarbons (such as butane or propane).

[0195] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of this invention to health. Such dosage forms can also be prepared by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0196] As used herein, the phrases "parenteral administration" and "administered parenterally" mean modes of administration other than enteral administration and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0197] Pharmaceutical compositions suitable for parenteral administration comprise a combination of one or more active compounds with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions immediately prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0198] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Appropriate fluidity can be maintained, for example, by the use of coating materials (such as lecithin), by maintaining the desired particle size in the case of dispersions and by the use of surfactants.

[0199] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. Preventing the action of microorganisms can be ensured by including various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents such as sugars, sodium chloride, etc. in the composition. In addition, delayed absorption of injectable drug forms may be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0200] In some cases, in order to prolong the effect of the drug, it is necessary to slow down the absorption of the drug of subcutaneous injection or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with low water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn can depend on crystal size and crystalline form. Alternatively, the delayed absorption of the drug form for parenteral administration can be achieved by dissolving or suspending the drug in an oil vehicle.

[0201] Injectable reservoir forms are prepared by forming a microencapsulated matrix of the subject compound in biodegradable polymers such as polylactide-polyglycolide. According to the ratio of drug to polymer and the property of the specific polymer adopted, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Reservoir injectable formulations are also prepared by trapping the drug in liposomes or microemulsions compatible with body tissues.

[0202] The method of introduction can also be provided by a rechargeable or biodegradable device. In recent years, various slow-release polymer devices have been developed and tested in vivo to control the delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form implants to slowly release the compounds of the compositions of the present invention at specific target sites.

[0203] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0204] The selected dosage level will depend upon a variety of factors including the activity of the particular combination of compounds employed, or the esters, salts or amides thereof, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0205] In general, a suitable daily dose of the active compound used in the compositions and methods of the invention will be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0206] The patient receiving such treatment is any animal in need thereof, including primates, particularly humans, and other mammals such as horses, cattle, pigs, and sheep; and poultry and pets in general.

[0207] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0208] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0209] In certain embodiments, the compositions described herein enhance the macrophage phagocytic activity of cancer cells, such as AML cells. In other embodiments, the phagocytic activity is enhanced by, for example, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% relative to the macrophage in the absence of the compositions described herein.

[0210] In certain embodiments, the present invention provides use of a composition of the present invention in the preparation of a medicament.

[0211] In certain embodiments, the present invention provides use of a composition of the present invention in the preparation of a medicament, for example, for the treatment of cancer.

[0212] In certain embodiments, the present invention provides methods for treating cancer, wherein the method comprises administering to a subject in need thereof, eg, a therapeutically effective amount of a composition of the present invention.

[0213] In certain embodiments, the present invention provides methods of inhibiting tumor cell growth and / or metastasis by administering to a subject in need thereof, for example, a therapeutically effective amount of a composition of the present invention.

[0214] Representative tumor cells include cells of cancers such as, but not limited to, melanoma, renal cancer, prostate cancer, breast cancer, colon cancer and lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, malignant melanoma of the skin or in the eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, cancer of the endocrine system, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer , penile cancer, chronic or acute leukemias including (acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia), solid tumors of childhood, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) tumors, non-small cell lung cancer (NSCLC), primary CNS lymphoma, tumor angiogenesis, spinal tumors, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, B-cell lymphoma, myeloproliferative disorders / neoplasms (MPDS), myelodysplastic syndrome, giant cell myeloma, heavy chain myeloma, light chain myeloma and Bence-Jones myeloma, cancers caused by the environment including cancers caused by asbestos (e.g., mesothelioma), and combinations of the aforementioned cancers. The term "treatment" includes preventive and / or therapeutic treatment. The term "preventive or therapeutic" treatment is recognized in the art and includes administering one or more of the subject compositions to the host. If administered before clinical manifestation of an undesirable symptom (e.g., a disease or other undesirable state of a host animal), the treatment is preventive (i.e., it protects the host from developing an undesirable symptom), and if administered after manifestation of an undesirable symptom, the treatment is therapeutic (i.e., it is intended to attenuate, improve or stabilize an existing undesirable symptom or its side effect).

[0215] As used herein, the term 'compound' includes compounds of Formula (I), (IA), (IB), (IC), (ID), (IE), (IF) and pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0216] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5-7 ring, more preferably a 6-ring. The term "aryl" also includes a polycyclic system with two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is aromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic. Aryl groups include benzene, naphthalene, phenanthrene, etc. Preferably, the term 'aryl' includes phenyl.

[0217] The term "heteroaryl" includes substituted or unsubstituted aromatic monocyclic structures, preferably 5 to 7 rings, more preferably 5 to 6 rings, whose ring structure includes at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic systems with two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is heteroaromatic, for example, other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic radicals. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, indole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole, benzimidazole, pyrimidine, etc. The heteroaryl group can be substituted by any optional substituent described herein at one or more positions where valence allows. Preferably, the term 'heteroaryl' includes imidazolyl and indolyl.

[0218] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the incidence of the disorder or condition in treated samples relative to untreated control samples, or delays the onset of or reduces the severity of one or more symptoms of the disorder or condition relative to untreated control samples.

[0219] The term "treatment" includes preventive and / or therapeutic treatment. The term "preventive or therapeutic" treatment is recognized in the art and includes administering one or more of the subject compositions to the host. If administered before clinical manifestation of an undesirable symptom (e.g., a disease or other undesirable state of a host animal), the treatment is preventive (i.e., it protects the host from developing an undesirable symptom), and if administered after manifestation of an undesirable symptom, the treatment is therapeutic (i.e., it is intended to attenuate, improve or stabilize an existing undesirable symptom or its side effect).

[0220] As used herein, the phrase "delay progression" refers to a procedure or application intended to delay the development of a disease or symptoms of a disease in time, including delaying the appearance or occurrence of at least one symptom of a particular disease in time.

[0221] The term "prodrug" is intended to encompass compounds that are converted into therapeutic agents of the present invention (e.g., compounds of formula (I)) under physiological conditions. Common methods for preparing prodrugs include one or more selected parts that are hydrolyzed under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of formula (I) in the formulations represented above may be replaced with corresponding suitable prodrugs, such as where the hydroxyl group in the parent compound exists in the form of an ester, or the carbonate or carboxylic acid present in the parent compound exists in the form of an ester.

[0222] As used herein, the terms "comprise" or "comprising" generally mean including, that is, allowing for the presence of one or more additional (unspecified) features or components.

[0223] As used herein, use of the term "including," as well as other forms such as "include," "includes," and "included," is not limiting.

[0224] As used herein, the term "disease" or "disorder" refers to a pathological condition in an organism caused by a cause or condition (including but not limited to infection, acquired conditions, genetic conditions) and characterized by recognizable symptoms.

[0225] As used herein, a "patient" or "subject" or "individual" to be treated includes humans and / or non-human animals, including mammals. Mammals include primates, such as humans, chimpanzees, gorillas, and monkeys; domesticated animals, such as dogs, horses, cats, pigs, goats, cows; and rodents, such as mice, rats, hamsters, and gerbils.

[0226] The term CD47+ disease cell refers to a cell having a CD47+ phenotype and associated with a disease. In one embodiment, the CD47+ disease cell is a cancer cell.

[0227] The term "CD47+" is used to refer to the phenotype of cells targeted by the binding of the CD47-SIRPα blocker of the present invention. CD47+ cells can be identified by flow cytometry using CD47 antibodies as affinity ligands. Appropriately labeled CD47 antibodies are commercially available for this purpose (e.g., the antibody product of clone B6H12 can be purchased from Santa Cruz Biotechnology). Cells examined for the CD47 phenotype may include standard tumor biopsy samples, especially blood samples collected from subjects suspected of carrying endogenous CD47+ cancer cells. CD47 disease cells that are particularly interesting as targets for treatment with the drug combination of the present invention are those that "overexpress" CD47. These CD47+ cells are typically disease cells, and the CD47 density on their surface exceeds the normal CD47 density of a given type of cell. CD47 overexpression will vary between different cell types, but in this article it means that any CD47 level determined by, for example, flow cytometry or by immunostaining or by gene expression analysis is greater than the measurable level on a healthy corresponding cell with a normal CD47 phenotype for that cell type.

[0228] The present invention includes compositions comprising pharmaceutically acceptable salts of compounds described herein and their uses in the compositions and methods of the present invention. In certain embodiments, expected salts include, but are not limited to, alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, expected salts of the present invention include, but are not limited to, L-arginine, benthamine, benzathine penicillin, betaine, calcium hydroxide, choline, danol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hyaluronic acid salts, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, expected salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts.

[0229] Pharmaceutically acceptable acid addition salts may also exist in the form of various solvates, such as solvates with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or insoluble in such solvents.

[0230] As used herein, the term "pharmaceutically acceptable salts" is intended to include all salts known and used in the pharmaceutical art. Pharmaceutically acceptable salts include, but are not limited to, amine salts such as, but not limited to, chloroprocaine, choline, N,N'-dibenzyl-ethylenediamine, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methylglucamine, procaine, N-benzyl-phenethylamine, 1-p-chloro-benzyl-2-pyrrolidin-1'-ylmethyl-benzimidazole, diethylamine and other alkylamines, piperazine, and tris(hydroxymethyl)aminomethane; alkali metal salts such as, but not limited to, lithium, potassium, and sodium; alkaline earth metal salts such as, but not limited to, barium, calcium, and magnesium; transition metal salts such as, but not limited to, zinc; and other metal salts such as, but not limited to, sodium hydrogen phosphate and disodium phosphate; and also include, but are not limited to, salts of inorganic acids such as, but not limited to, hydrochlorides and sulfates; and salts of organic acids such as, but not limited to, acetates, lactates, malates, tartrates, citrates, ascorbates, succinates, butyrates, valerates, and fumarates. Exemplary pharmaceutically acceptable salts include acetate, lactobionate, benzenesulfonate, laurate, benzoate, malate, bicarbonate, maleate, bisulfate, mandelate, bitartrate, methanesulfonate, borate, methyl bromide, methylnitrate, calcium edetate, methylsulfate, camphorsulfonate, mucate, carbonate, naphthenate, bromide, chloride, nitrate, clavulanate, N-methylglucamine, citrate, ammonium, dihydrochloride, oleate, edetate, oxalate, edisylate, pamoate (pamoate), etopoate, palmitate, estolate, The pharmaceutically acceptable salts of the present invention are hydroxybenzoate, hydroxynaphtho ...

[0231] The term "stereoisomer" refers to any enantiomer, diastereomer or geometric isomer of a compound such as described herein. When such compounds are chiral, they may exist in racemic or optically active form. Since the pharmaceutical activity of the racemates or stereoisomers of the compounds according to the present invention may be different, it may be necessary to use a compound enriched in one enantiomer. In these cases, the final product or even the intermediate can be separated into enantiomeric compounds by chemical or physical measures known to those skilled in the art, or even used in synthesis for this purpose. In the case of racemic amines, diastereomers are formed from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulfonic acids in R and S forms. Also advantageous is the chromatographic enantiomeric resolution with the aid of optically active resolving agents such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel.

[0232] As used herein, the term "ester" refers to the group -C(O)OR 11 , where R 11 represents a hydrocarbon group.

[0233] As used herein, the term "amide" refers to the group -C(O)NH2.

[0234] In certain embodiments, the compounds of the present invention may be racemic. In certain embodiments, the compounds of the present invention may be enriched in one enantiomer. For example, the compounds of the present invention may have greater than 30% ee, 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee or even 95% or higher ee. In certain embodiments, the compounds of the present invention may have more than one stereocenter. In certain such embodiments, the compounds of the present invention may be enriched in one or more diastereomers. For example, the compounds of the present invention may have greater than 30% de, 40% de, 50% de, 60% de, 70% de, 80% de, 90% de or even 95% or higher de.

[0235] The term "subject" includes mammals (particularly humans) and other animals, such as domesticated animals (eg, household pets including cats and dogs) and non-domesticated animals (such as wild animals).

[0236] In certain embodiments, the present invention provides a composition comprising a small molecule CD-47-SIRPα pathway inhibitor and an agent capable of activating receptors such as Fc receptors (FcR) or pro-phagocytic receptors or other therapeutic modalities for activating pro-phagocytic receptors in cancer therapy to maximize the potential of CD-47-SIRPα pathway blockade.

[0237] Example-1: The synthetic procedure for preparing the compounds described in the present invention is described in the co-pending Indian Provisional Patent Application No. 201841001438 filed on January 12, 2018, which was converted into PCT Application No. PCT / IB2019 / 050219, the contents of which are incorporated herein by reference in their entirety.

[0238] Biological Examples:

[0239] Reagents DPBS (Gibco), RPMI 1640 with HEPES and L-GLN-500ML (Lonza), recombinant human M-CSF (R&D systems), CD47 monoclonal antibody (B6H12), functional grade antibody (Ebioscience), functional grade mouse IgG1κ isotype control (Ebioscience), Vacutainer tubes (multiple sample Luer connector) (BD), Vacutainer tubes (heparin sodium (NH) 158 USP units), blood collection tubes (BD), Histopaque (density 1.077 gm / ml) (SIGMA 1077), Trypan blue solution (SIGMA-T8154), blood cell counter (Bright line-SIGMA Z359629), scalp vein infusion set (JMS), cell dissociation buffer (Gibco), 48-well sterile flat bottom plates (Corning), Raji cells expressing luciferase (produced in-house by transfecting Raji cells with luciferase gene) luminometer, hygromycin B (Invitrogen), Bright Glo luciferase assay system (Promega), 96-well plate, polystyrene, high band, white flat-bottom wells (Sigma CLS3912), anti-human CD20 antibody (Invivogen hcd20-mab1), bortezomib (Selleckchem, S1013)

[0240] Example-2: Luciferase-based phagocytosis assay

[0241] In vitro phagocytosis assays were performed to evaluate the ability of the test items to enhance macrophage phagocytic activity. Monocytes were isolated from the blood of healthy donors and cultured for 6-8 days using 10% RPMI (Roswell Park Memorial Institute) medium and recombinant human M-CSF to differentiate into macrophages. The culture medium was changed every other day. After differentiation, adherent macrophages were collected by gentle scraping and cultured overnight in 48-well tissue culture plates at a density of 100,000 per well in 10% RPMI. At the same time, Raji (lymphoma cell line) cells expressing luciferase were cultured in 10% RPMI medium with 100 μg / mL hygromycin B in tissue culture flasks. On the day of phagocytosis, macrophages were serum starved for 2 hours. 400,000 luciferase-expressing Raji cells per well were incubated with anti-human CD47 antibody or mouse IgG1 K isotype control antibody or various concentrations of selected compounds of the present invention alone or in combination with anti-human CD20 antibody in serum-free medium for 30 minutes at 37°C and added to each well of a 48-well plate seeded with macrophages. After 2 hours, the cells were washed twice with PBS and 100 μl of serum-free RPMI was added to each well. In addition, 50 μl of bright glow reagent was added to each well, and the cells were then mixed and incubated in the dark for 5 minutes. After transferring the contents of each well to a white plate, the luminescence readings were read using a microplate reader. The luminescence intensity indicates the extent of phagocytosis. Each experimental condition was performed in duplicate. The results are shown in Figure 1 Shown in.

[0242] Figure 1 The results in show that treatment of tumor cells with anti-human CD20 antibodies resulted in a significant increase in phagocytosis compared to CD47-SIRPα blocking compounds alone.

[0243] Example-3: Human macrophage phagocytosis assay to evaluate compounds in combination with bortezomib

[0244] In vitro phagocytosis assays were performed to evaluate the ability of the test items to enhance the phagocytic activity of macrophages. Monocytes were isolated from the blood of healthy donors and cultured for 6-8 days using complete RPMI (Roswell Park Memorial Institute) medium supplemented with 20 ng / mL recombinant human M-CSF to differentiate them into macrophages. The culture medium was changed every other day. H929 cells were cultured in complete RPMI medium at the same time and treated with 10 nM bortezomib for 48 hours. Macrophages were starved in serum-free RPMI for 2 hours. CFSE-stained, bortezomib-treated / untreated H929 (multiple myeloma cell line) cells were incubated at 0.2 x 10 6The cells were seeded at a density of 10 cells / well in a 96-well low attachment plate and treated with anti-human CD47 purified B6H12 (5 μg / mL) / mouse IgG1 K isotype control antibody (5 μg / mL) / different concentrations of selected compound 6 of the present invention in serum-free medium for 30 minutes at 37°C. Serum-starved macrophages were added to H929 cells at a ratio of 1:4 (macrophages: H929 cells) and incubated at 37°C for 2 hours. The cells in each well were stained with anti-human CD11b APC at 4°C in the dark for 30 minutes, followed by fixation and FACS analysis. FITC and APC-positive cells were considered to be H929 cells phagocytosed by macrophages. The data acquired using FACS verse was analyzed using Flow Jo software. The results are in Figure 2 Shown in.

[0245] Figure 2 Results in 2014 showed that treatment of tumor cells with a proteasome inhibitor (bortezomib) resulted in a significant increase in phagocytosis compared to CD47-SIRPα blocking compounds alone.

[0246] Example-4: Efficacy study of the combination of compound 6 and PD-L1 antibody in the A20 syngeneic lymphoma model

[0247] Female Balb / c (BALB / cAnNTac) mice (6-8 weeks old) bred in-house were used for this efficacy study in the A20 syngeneic lymphoma model. Animals were individually tail-tagged and caged with cage cards indicating the study code, experimental date, sex, and number of animals. Animals were weighed daily during the experiment. The A20 cell line (a B-cell lymphoma line derived from spontaneous reticulum neoplasms of old BALB / cAnN mice) was purchased from ATCC.

[0248] When the average tumor volume reaches about 75 mm 3 At 4 hr, the animals were randomly divided into four groups (G1 to G4) according to the tumor volume, with twelve animals in each group (N=12), and were administered with vehicle, compound 6, anti-mouse PD-L1 antibody, and a combination of compound 6 and anti-mouse PD-L1 antibody as described below:

[0249]

[0250] Treatment lasted for 21 days, during which time overall efficacy and tolerability were assessed based on changes in tumor volume and body weight observed during treatment.

[0251] The body weight of each animal was recorded daily before administration of compound 6 throughout the experiment. The mortality / morbidity of the animals was observed once a day throughout the experiment, and the clinical signs of the animals were observed once a day throughout the experiment. The tumor volume of all animals in the treatment groups was measured three times a week (once every 2-3 days) using a digital vernier caliper. As a measure of efficacy, the values ​​of T (treatment) / C (control)% and TGI (tumor growth inhibition%)% were calculated. GraphPad was used to analyze the tumor volume of all animals in the treatment group. Version 7.0 was used for graphics and statistical analysis. To analyze tumor volume data, statistical comparisons were performed on all groups using one-way ANOVA and Dunnett's multiple comparison test on day 18. All analyses and comparisons were evaluated at the 5% (p<0.05) level. "p" values ​​less than 0.05 were considered significant. The results and statistical data are summarized in the table below.

[0252]

[0253] One-way ANOVA, Dunnett's multiple comparison test: *-p<0.05, ***-p<0.001; TGI-tumor growth inhibition;

[0254] Compound 6 was well tolerated at 30 mg / kg bid, both as a single agent and in combination with PD-L1 antibody, without any treatment-related clinical signs and mortality, indicating that the test drug was extremely well tolerated at the administered dose. At the end of the treatment period, compound 6 administered alone at 30 mg / kg, anti-mouse PD-L1 antibody alone, and compound 6 in combination with anti-mouse PD-L1 showed tumor growth inhibition (TGI) values ​​of 72%, 41%, and 90%, respectively. The effect of treatment on tumor growth kinetics is graphically represented in Figure 3 middle.

[0255] It was further observed that the combination of compound 6 and anti-mouse PD-L1 antibody significantly enhanced tumor growth inhibition and more durable responses when compared to individual treatments. Five of the 11 animals treated with the combination of compound 6 and anti-mouse PD-L1 antibody showed complete tumor regression. The tumor growth inhibition observed with compound 6 alone and compound 6 in combination with anti-mouse PD-L1 antibody was statistically significant.

Claims

1. A combination product comprising a CD47-SIRPα blocker and one or more anticancer agents, wherein the CD47-SIRPα blocker is selected from: and , or a pharmaceutically acceptable salt thereof, wherein the anticancer agent is selected from anti-CD20 antibodies, proteasome inhibitors, antibodies to PD-1 and antibodies to PD-L1, and the proteasome inhibitor is selected from bortezomib, ixazomib and carfilzomib.

2. The combination product according to claim 1, wherein the anti-cancer agent is an anti-CD20 antibody selected from rituximab, tezetamipan and tositumomab.

3. The combination product according to claim 1, wherein the anticancer agent is bortezomib.

4. The combination product according to claim 1, wherein the antibody against PD-1 and the antibody against PD-L1 are selected from nivolumab, pembrolizumab, atezolizumab, durvalumab and camrelizumab. 5 . The combination product according to claim 1 , wherein the CD47-SIRPα blocker is a substance that blocks the interaction between CD47 and SIRPα.

6. The combination product according to claim 5, wherein blocking the interaction between CD47 and SIRPα induces macrophages to phagocytose tumor cells expressing CD47.

7. Use of a combination comprising a CD47-SIRPα blocker and one or more anticancer agents in the manufacture of a medicament for treating a disease or condition mediated by the CD47-SIRPα pathway in a subject or delaying the progression of the disease or condition, wherein the disease or condition mediated by the CD47-SIRPα pathway is cancer, wherein the CD47-SIRPα blocker is selected from: and , or a pharmaceutically acceptable salt thereof; wherein the anticancer agent is selected from anti-CD20 antibodies, proteasome inhibitors, antibodies to PD-1 and antibodies to PD-L1, and the proteasome inhibitor is selected from bortezomib, ixazomib and carfilzomib.

8. The use according to claim 7, wherein the anticancer agent is bortezomib.

9. The use according to claim 7, wherein the treatment with one or more anticancer agents is carried out with a and before, simultaneously or after treatment with a compound or a pharmaceutically acceptable salt thereof.

10. The method of claim 7 , wherein the cancer is selected from the group consisting of melanoma, kidney cancer, prostate cancer, breast cancer, colon cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, malignant melanoma of the skin or in the eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, cancer of the endocrine system, thyroid cancer. carcinoma, parathyroid carcinoma, adrenal carcinoma, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, solid tumors of childhood, lymphocytic lymphoma, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) tumors, tumor angiogenesis, spinal tumors, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, B-cell lymphoma, multiple myeloma, cancers caused by the environment, and combinations of the above cancers.

11. The use according to claim 10, wherein the acute or chronic leukemia is selected from acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia and chronic lymphocytic leukemia.

12. The use according to claim 10, wherein the environmentally induced cancer is asbestos-induced cancer.

13. The use according to claim 10, wherein the cancer is mesothelioma.

14. The use according to claim 10, wherein the cancer is selected from non-small cell lung cancer (NSCLC) and primary CNS lymphoma.

15. A pharmaceutical kit comprising a combination product according to any one of claims 1 to 6 and a package insert comprising instructions for administration of the drug for treating a subject presenting with a CD47 pathway disorder. 16 . A pharmaceutical composition comprising the CD47-SIRPα blocker according to any one of claims 1 to 6 , one or more anticancer agents, and a pharmaceutically acceptable carrier.

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