Pharmaceutical composition comprising an HDAC inhibitor and an anti-PD1 antibody or an anti-PD-L1 antibody

Abstract

The present invention relates to a method for killing cancer cells using a combination of an HDAC inhibitor and an anti-PD-1 antibody or an anti-PD-L1 antibody. The present invention can effectively affect the following aspects: prolonging the survival of animals suffering from cancer, tumors, tumor-related diseases and tumor diseases; inhibiting the growth of tumor-related proliferating cells; or tumor degeneration.

Description

Technical Field

[0001] The present invention relates to a method for inhibiting cancer cell growth and killing cancer cells by combining an HDAC inhibitor with an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0002] The present application claims priority for the combined use of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivatives, or salts thereof with anti-PD-1 antibodies in cancer treatment, as the applicant has demonstrated in Korean Patent No. 1978364, entitled "PHARMACEUTICALLY ACCEPTABLE SALT OF ALKYLCARBAMOYL NAPHTHALENYLOXYOCTENOYLHYDROXYAMIDE OR OF DERIVATIVE THEREOF AND METHOD FOR PREPARING SAME," registered on May 8, 2019, that the specific immunoactivity of epigenetic target inhibitor alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivatives, or salts thereof has a synergistic effect in the anti-cancer effect of anti-PD-1 antibodies.

[0003] In addition, this application claims priority from Korean Patent Application No. 2019-0109256, filed on September 4, 2019, the entire contents of which are incorporated into this specification. Background Art

[0004] Histones are basic proteins that bind to DNA in the nucleus of eukaryotic cells, and at specific positions on each histone molecule, the amino group of the lysine residue undergoes reversible acetylation. Histone acetylation is associated with the formation of higher-order chromatin structures or the cell division cycle, and is therefore involved in the regulation of genetic information expression. It is also stably regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). It is understood that these enzymes neutralize the positive charge of the amino-terminal lysine residues of histones (the four residues of H4) by acetylation to induce transcriptional activity, or deacetylate them to provide charge again and inhibit transcription, thereby inducing a balance in the level of histone acetylation and regulating gene expression during the transcriptional stage.

[0005] HDACs have recently been shown to play a role in promoting cell proliferation by suppressing the expression of cell proliferation inhibitors through their high expression under adverse environmental conditions such as hypoxia, low glucose, and cell carcinogenesis. Therefore, they are considered important factors in regulating cell carcinogenicity and differentiation. In other words, if chromatin hyperacetylation inhibits cell proliferation and promotes differentiation, then HDACs play a crucial role in inducing cell proliferation through histone deacetylation. This is supported by the fact that treatment with HDAC inhibitors leads to the inhibition of cell proliferation and angiogenesis.

[0006] Meanwhile, PD-1 is known to play an important role in regulating immune responses and maintaining peripheral tolerance. PD-1 is moderately expressed on naive T cells, B cells, and NKT cells, and its expression is regulated by T / B cell receptor signaling on lymphocytes, monocytes, and myeloid cells.

[0007] PD-1 has two known ligands: PD-L1 (B7-H1) and PD-L2 (B7-DC), which are expressed on cancer cells from a variety of tissues. Specifically, in cells from various cancers such as ovarian cancer, kidney cancer, colorectal cancer, pancreatic cancer, liver cancer, and melanoma, PD-L1 expression is associated with poor prognosis and reduced overall survival, regardless of subsequent treatment.

[0008] Therefore, it is understood that the interaction of PD-L1 on tumor cells with PD-1 on T cells reduces T cell activation and decreases immune surveillance function, leading to an impaired immune response to tumors.

[0009] It is expected that the therapeutic efficacy of these antibodies can be further enhanced by combining them with radiotherapy, surgery, chemotherapy, targeted therapy, signaling pathway inhibitors, immune enhancers, etc.

[0010] Therefore, for effective application in the reduction of cancer cell growth and necrosis, research on combination therapy with suitable inhibitors and antibodies as described above is needed. Summary of the Invention

[0011] Technical issues

[0012] In order to solve the above problems, the present invention provides a method for killing cancer cells by combining an HDAC inhibitor with an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0013] Solution to the problem

[0014] The present invention provides a pharmaceutical composition comprising:

[0015] Alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof; and

[0016] Anti-PD-1 antibody or anti-PD-L1 antibody.

[0017] According to one embodiment, the weight ratio of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof to the anti-PD-1 antibody or anti-PD-L1 antibody may be 1:0.1 to 1:15.

[0018] According to one embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered at a dose of 1 to 200 mg / kg.

[0019] According to one embodiment, the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or its salt may be administered at a dose of 10 to 500 mg / kg.

[0020] According to one embodiment, the anti-PD-1 antibody may include one or more selected from pembrolizumab, nivolumab, camrelizumab, cemiplimab, sintilimab, and toripalimab.

[0021] According to one embodiment, the anti-PD-L1 antibody may include one or more selected from atezolizumab, avelumab, and durvalumab.

[0022] According to one embodiment, the alkylaminoformylnaphthyloxyoctenoylhydroxyamide or its derivatives can be one or more selected from the following compounds:

[0023] 1)(E)-N1-(3-(1H-imidazol-1-yl)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0024] 2) (E)-N8-hydroxy-N1-(4-hydroxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0025] 3)(E)-N1-(3-(dimethylamino)-2,2-dimethylpropyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0026] 4) (E)-N1-(2-(diisopropylamino)ethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0027] 5) (E)-N8-hydroxy-N1-(1-methoxyprop-2-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0028] 6) (E)-N8-hydroxy-N1-(4-methoxybenzyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0029] 7) (E)-N1-(4-fluorophenethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0030] 8) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(tetrahydrofuran-2-yl)methyl)-2-octenediamide,

[0031] 9) (E)-N1-(2-cyclohexenylethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0032] 10) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(3-(2-oxopyrrolidin-1-yl)propyl)-2-octenediamide,

[0033] 11)(E)-N1-(furan-2-ylmethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0034] 12) (E)-N1-(4-(dimethylamino)benzyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0035] 13) (E)-N8-hydroxy-N1-(2-methoxyethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0036] 14) (E)-N1-cyclohexyl-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0037] 15) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(thiophen-2-ylmethyl)-2-octenediamide,

[0038] 16) (E)-N8-hydroxy-N1-(4-methoxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0039] 17) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(4-(trifluoromethoxy)benzyl)-2-octenediamide,

[0040] 18) (E)-N1-(1-(cyclohexylmethyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0041] 19) (E)-N1-(1-cyclopentylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0042] 20)(E)-N1-(1-benzylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0043] 21)(E)-N8-hydroxy-N1-(1-isopropylpyrrolidin-3-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0044] 22) (E)-N1-(1-(cyclohexanecarbonyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0045] 23) (E)-tert-butyl 3-(8-(hydroxyamino)-2-((naphthalen-1-yloxy)methyl)-8-oxo-2-octenylamido)pyrrolidine-1-carboxylate,

[0046] 24) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(pyrrolidin-3-yl)-2-octenediamide,

[0047] 25) (E)-N1-(1-cyclohexylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-2-yloxy)methyl)-2-octenediamide,

[0048] 26) (E)-N1-(1-cyclopropylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0049] 27) (E)-N1-(1-cyclopropylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0050] 28) (E)-N1-(1-ethylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0051] 29) (E)-N1-(1-ethylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0052] 30) (E)-N8-hydroxy-N1-(2-(1-methylpyrrolidin-2-yl)ethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0053] 31)(E)-N8-hydroxy-N1-(1-isopropylpiperidin-4-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, and

[0054] 32)(E)-N1-(3-(Dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide.

[0055] According to one embodiment, the salt of the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide may be (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide phosphate.

[0056] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof; and

[0057] The anti-PD-1 antibody or anti-PD-L1 antibody may be administered simultaneously as a single formulation, or may be administered simultaneously or sequentially or in reverse order as separate formulations.

[0058] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof; and

[0059] The anti-PD-1 antibody or anti-PD-L1 antibody can be prepared as a single formulation.

[0060] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof; and

[0061] The anti-PD-1 antibody or anti-PD-L1 antibody can be prepared in separate compositions.

[0062] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof; and

[0063] The anti-PD-1 antibody or anti-PD-L1 antibody can be administered at a dose less than or equal to the therapeutically effective dose, respectively.

[0064] According to one embodiment, 1) alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof is administered before administering anti-PD-1 antibody or anti-PD-L1 antibody, or

[0065] 2) Anti-PD-1 antibody or anti-PD-L1 antibody can be administered before administration of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof.

[0066] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof and anti-PD-1 antibody or anti-PD-L1 antibody can be formulated into separate preparations, so that alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof is administered to an animal intravenously or orally, and anti-PD-1 antibody or anti-PD-L1 antibody is administered to an animal intravenously.

[0067] According to one embodiment, the composition can synergistically inhibit the growth of cancer cells or kill cancer cells.

[0068] According to one embodiment, the cancer cells may be characterized by the expression of PD-L1, and the cancer cells include hepatitis A virus-induced liver cancer cells, hepatitis B virus-induced liver cancer cells, hepatitis C virus-induced liver cancer cells, non-virus-related liver cancer cells, metastatic liver cancer cells, colon cancer cells, pancreatic cancer cells, blood cancer cells, melanoma cells or lung cancer cells.

[0069] According to one embodiment, cancer cells can be screened by a method comprising measuring PD-L1 expression in tumor tissue.

[0070] According to one embodiment, the PD-L1 tumor proportion score (TPS) on cancer cells may be 1% or higher.

[0071] Specific details of other embodiments of the present invention are included in the following detailed description.

[0072] Effects of the Invention

[0073] The method for killing cancer cells by combining an HDAC inhibitor with an anti-PD-1 antibody or an anti-PD-L1 antibody according to the present invention can effectively kill cancer cells.

[0074] Cancer, tumors, tumor-related diseases, and neoplastic diseases are generally life-threatening diseases characterized by rapid cell proliferation. The present invention can effectively affect the following aspects: prolonging the survival of animals suffering from such diseases or conditions; inhibiting the growth of tumor-related proliferating cells; or tumor degeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] Figure 1 and 2 Shown are the tumor growth inhibition rates in mouse models. DETAILED DESCRIPTION

[0076] Because the present invention is susceptible to various modifications and variations, specific embodiments are shown in the accompanying drawings and will be described in detail in the detailed description. However, it should be understood that the present invention is not limited to the specific embodiments, but encompasses all modifications, equivalents, and alternatives within the spirit and scope of the present invention. In the following description of the present invention, detailed descriptions of known functions will be omitted if it is determined that they may obscure the main purpose of the present invention.

[0077] The following describes in more detail the method for killing cancer cells using a combination of an HDAC inhibitor and an anti-PD-1 antibody or an anti-PD-L1 antibody according to an embodiment of the present invention.

[0078] As used herein, the term "abnormal cell growth" refers to cell growth that is independent of normal regulatory mechanisms, including abnormal growth of normal cells and abnormal cell growth, such as loss of contact inhibition. That is, when cells come into contact with adjacent cells or tissues during growth, the mechanisms that inhibit cell growth and division are lost.

[0079] As used herein, the term "neoplasia" refers to abnormal, unregulated, and disorganized cell proliferation that is distinguished from normal cells by autonomous growth and somatic mutation. As tumor cells grow and divide, they pass on their genetic mutations and proliferation characteristics to progeny cells. A tumor or neoplasm is an accumulation of tumor cells. In some embodiments, tumors can be benign or malignant.

[0080] As used herein, the term "metastasis" refers to the spread of tumor cells through lymphatic or vascular channels. Metastasis also refers to the migration of tumor cells through direct extension through the serous or subarachnoid spaces or other spaces. Through the process of metastasis, tumor cells migrate to other parts of the body and form tumors in areas distant from the site of initial appearance.

[0081] As used herein, the term "subject" refers to an animal, including but not limited to a primate (e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject" and "patient" are used interchangeably herein to refer to, for example, a mammalian subject, such as a human subject.

[0082] As used herein, the terms "treat," "treatment," and "therapy" are meant to include alleviating or eliminating a disorder, disease, or condition; or one or more symptoms associated with a disorder, disease, or condition; or alleviating or eradicating the cause of the disorder, disease, or condition itself.

[0083] As used herein, the term "therapeutically effective dose" refers to a dose of a compound that, when administered, is sufficient to prevent the development of, or alleviate to some extent, one or more symptoms of the disorder, disease, or condition being treated. The term "therapeutically effective dose" also refers to a dose of a compound that is sufficient to elicit the biological or medical response in a cell, tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or clinician.

[0084] As used herein, the term "pharmaceutical composition" is used interchangeably with "pharmaceutical composition" and "pharmaceutically acceptable composition" and refers to a composition that is relatively non-toxic and has a non-injurious, effective effect on the subject to which it is administered. Furthermore, it may refer to any formulation of an organic or inorganic compound that produces side effects that do not impair the efficacy of the drug, does not cause severe irritation to the subject to which the compound is administered, and does not diminish the biological activity and properties of the compound.

[0085] As used herein, the terms "pharmaceutically acceptable carrier," "pharmaceutically acceptable excipient," "physiologically acceptable carrier," or "physiologically acceptable excipient" refer to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each component must also be "pharmaceutically acceptable" in terms of compatibility with the other ingredients of the pharmaceutical formulation. It must also be suitable for use in contact with human and animal tissues or organs, without excessive toxicity, irritation, allergic reaction, immunogenicity, or other problems or complications, and commensurate with a reasonable benefit / risk ratio. See Remington: The Science and Practice of Pharmacy, 21st Edition; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 5th Edition; Rowe et al, Eds., The Pharmaceutical Press and the American Pharmaceutical Association: 2005; and Handbook of 'Pharmaceutical Additives, 3rd Edition; Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, Gibson Ed., CRC Press LLC: Boca Raton, FL, 2004.

[0086] The term "pharmaceutical composition" as used herein refers to a mixture of the compound disclosed herein and other chemical components, such as pharmaceutically acceptable diluents, carriers, etc. The compound can be easily administered to an organism via a pharmaceutical composition.

[0087] In the present invention, the various component drugs of the pharmaceutical composition may be present as separate preparations or a single preparation, and these component drugs may be administered simultaneously, sequentially, or in reverse order. In addition, in the present invention, the pharmaceutically effective dose, administration time, administration interval, administration route, treatment period, etc. of the various component drugs of the pharmaceutical composition may be the same or different from each other.

[0088] As used herein, the term "administered subject" is used interchangeably with "administered individual" and "administered organism" and refers to all animals, including humans, that have been infected or can be injected with bacteria or drug-resistant strains.

[0089] The term "death" as used herein includes both necrosis and apoptosis.

[0090] As discussed here, "angiogenesis" is important in the development and metastasis of tumors. Angiogenic factors have been found to be associated with several solid tumors, such as rhabdomyosarcoma, retinoblastoma, Ewing's sarcoma, neuroblastoma, and osteosarcoma. Without a blood supply to provide nutrients and remove cellular waste, tumors cannot expand. Tumors for which angiogenesis is important include solid tumors such as renal cell carcinoma and hepatocellular carcinoma, and benign tumors such as acoustic neuroma and neurofibromatosis. Angiogenesis is associated with blood-borne tumors such as leukemia. It is believed that angiogenesis may play a role in the bone marrow abnormalities that lead to leukemia. Preventing angiogenesis can prevent the growth of cancerous tumors and the damage that the tumor's presence causes to the subject.

[0091] The present invention relates to a pharmaceutical composition and method for inhibiting the growth of cancer cells and killing cancer cells by combining an HDAC inhibitor with an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0092] HDAC is a family that includes at least 18 enzymes and is divided into three classes (classes I, II and III). Class I HDAC includes but is not limited to HDAC 1, 2, 3 and 8. Class I HDAC can be found in the nucleus and is thought to be related to transcriptional control repressors. Class II HDAC includes but is not limited to HDAC 4, 5, 6, 7 and 9, which can be found in the cytoplasm and nucleus. Class III HDAC is considered to be NAD-dependent protein, including but not limited to members of the sirtuin protein family. Non-limiting examples of sirtuin proteins include SIRT1-7. The term "selective HDAC" as used herein refers to an HDAC inhibitor that does not interact with all three classes of HDAC.

[0093] HDAC inhibitors are a class of therapeutic agents that promote differentiation and apoptosis in hematological and solid malignancies through chromatin remodeling and gene expression regulation. Several HDAC inhibitors have been identified, including benzamides (entinostat), short-chain fatty acids (i.e., sodium phenylbutyrate); hydroxamic acids (i.e., vorinostat and trichostatin A); a cyclic tetrapeptide containing a 2-amino-8-oxo-9,10-epoxy-decanoyl moiety (i.e., Trapoxin A); and a cyclic peptide without a 2-amino-8-oxo-9,10-epoxy-decanoyl moiety (i.e., FK228).

[0094] HDAC inhibitors can be broadly divided into pan-HDAC inhibitors and selective HDAC inhibitors. Although known HDAC inhibitors have great structural diversity, they share common features: a portion that interacts with the enzyme active site and a side chain located within the channel leading to the active site. This can be seen in hydroxamates such as SAHA (suberoylanilide hydroxamic acid), where the hydroxamate group is thought to interact with the active site. In the case of depsipeptides, it is believed that intracellular reduction of the disulfide bond generates a free thiol group (which interacts with the active site) connected to the 4-carbon olefin chain.

[0095] The various HDAC inhibitors differ in how they interact with the rim of the HDAC channel, which is located at the end of the channel opposite the active site. This interaction between the HDAC inhibitor and the channel rim is thought to explain, at least in part, some of the differences in HDAC selectivity observed between pan-HDAC inhibitors (such as SAHA) and selective HDAC inhibitors (such as depsipeptides).

[0096] Alkylcarbamoylnaphthyloctenoylhydroxyamide is an HDAC inhibitor that is being clinically investigated for use in multiple types of solid tumors and hematological cancers. Alkylcarbamoylnaphthyloctenoylhydroxyamide is rapidly absorbed with a half-life of approximately 7 to 8 hours, and importantly, the changes in histone acetylation persist for several days after administration.

[0097] Programmed cell death-1 (PD-1) is a cell surface receptor and a member of the CD28 family of T cell regulators within the immunoglobulin superfamily of receptors. The human PD-1 gene is located on chromosome 2q37. The full-length PD-1 cDNA encodes a 288-amino acid protein with 60% homology to murine PD-1. It is present on CD4-CD8- (double-negative) thymocytes during thymic development and is expressed upon activation in mature hematopoietic cells (such as T and B cells, NKT cells, and monocytes) following prolonged antigen exposure. PD-L1 has recently been shown to be expressed on many mouse and human tumors (and can be induced by IFNγ on most PD-L1-negative tumor cell lines) and is postulated to mediate immune evasion (Iwai Y. et al., Proc. Natl. Acad. Sci. USA 99: 12293-12297 (2002); Strome SE et al., Cancer Res., 63: 6501-6505 (2003). In humans, expression of PD-1 and / or PD-L1 has been found in many primary tumor biopsies from lung, liver, ovarian, cervical, skin, colon, glioma, bladder, breast, kidney, esophageal, gastric, oral squamous cell, urothelial and pancreatic cancers, as well as head and neck tumors (Brown JA et al., J. Immunol. 170: 1257-1266 (2003); Dong H. et al., J. Immunol. 171: 1267-1279 (2003); al., Nat. Med. 8:793-800 (2002); Wintterle et al., Cancer Res. 63:7462-7467 (2003); Strome SE et al., Cancer Res., 63:6501-6505 (2003); Thompson RH et al., Cancer Res. 66:3381-5 (2006); Thompson et al., Clin. Cancer Res. 13:1757-61 (2007); Nomi T. et al., Clin. Cancer Res. 13:2151-7. (2007)). PD-ligand expression on tumor cells is associated with poor prognosis in cancer patients with various tumor types (reviewed by Okazaki and Honjo, Int. Immunol. 19:813-824 (2007)).

[0098] In addition, high expression of PD-L1 on tumor cells is known to be associated with poor prognosis and survival in various other solid tumors. The PD-1 / PD-L1 pathway is expected to play a key role in tumor immune evasion and may be considered an attractive target for therapeutic intervention in several solid organ systems. Several PD-1 and PD-L1 antibodies are in clinical development and, overall, are reported to be well tolerated, with most experiencing no dose-limiting toxicity in Phase I studies. Multiple studies have shown that the interaction of PD-1 with its ligands (PD-L1 and PD-L2) leads to inhibition of lymphocyte proliferation both in vitro and in vivo. Therefore, binding of the ligand PD-L1 to PD-1 is expected to downregulate effector anti-tumor T cell activity and promote immune evasion.

[0099] Furthermore, disruption of the PD-1 / PD-L1 interaction has been shown to increase T cell proliferation and cytokine production and arrest cell cycle progression. In vitro studies using PD-1-specific antibodies to block PD-1 have demonstrated enhanced cytotoxic T cell responses to liver cancer-specific antigens, including an increase in the frequency of antigen-specific cells secreting IFN-γ. However, targeting PD-1 is expected to be an effective therapeutic strategy for cancer.

[0100] The main approach to targeting PD-1 is to develop genetically engineered monoclonal antibodies that inhibit PD-1 function by interfering with the binding of PD-1 to PD-L1. Anti-PD-1 antibodies and their antigen-binding portions bind to PD-1 with high specificity and affinity, blocking the binding of PD-L1 and / or PD-L2, and inhibiting the immunosuppressive effects of the PD-1 signaling pathway. In some embodiments, the combination therapy comprises administering alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivatives, or salts thereof, and an anti-PD-1 antibody or its antigen-binding portion, wherein the antibody or its antigen-binding portion is a chimeric, humanized, or human monoclonal antibody or a portion thereof. In certain embodiments, the anti-PD-1 antibody or its antigen-binding portion is a humanized antibody. In some embodiments, the anti-PD-1 antibody or its antigen-binding portion is a human antibody. In some embodiments, the anti-PD-1 antibody or its antigen-binding portion is a monoclonal antibody or its antigen-binding portion.

[0101] According to one embodiment, the HDAC inhibitor may include an alkylaminoformyl naphthoxyoctenoyl hydroxyamide, a derivative thereof, or a salt thereof. The alkylaminoformyl naphthoxyoctenoyl hydroxyamide is represented by Formula 1:

[0102] [Formula 1]

[0103]

[0104] Wherein R1 is unsubstituted or substituted C 1-3 Alkyl; unsubstituted or C 3-8 Cycloalkyl, C3-8 Cycloalkyl C 1-3 Alkyl, benzyl, C 1-3 Alkyl or C 3-8 Cycloalkylcarbonyl-substituted pyrrolidine; 1-3 Alkyl or C 3-8 Cycloalkyl-substituted piperidine; furan; or C 3-8 Cycloalkyl,

[0105] Provided that it does not include unsubstituted C 1-2 Alkyl and C 1-2 Alkylpyrrolidinyl substituted C 1-2 alkyl,

[0106] The salt may be selected from phosphate, tartrate, stearate, gluconate, fumarate, naphthoate, 1-hydroxy-2-salt and mixtures thereof, such as phosphate.

[0107] According to one embodiment, the salt may be selected from phosphates, tartrates, and mixtures thereof, which have relatively high stability and water solubility, and may include, for example, phosphates.

[0108] Preferred compounds as alkylcarbamoylnaphthyloxyoctenoylhydroxyamides of Formula 1 or derivatives thereof may be selected from the following compounds:

[0109] 1)(E)-N1-(3-(1H-imidazol-1-yl)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0110] 2) (E)-N8-hydroxy-N1-(4-hydroxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0111] 3)(E)-N1-(3-(dimethylamino)-2,2-dimethylpropyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0112] 4) (E)-N1-(2-(diisopropylamino)ethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0113] 5) (E)-N8-hydroxy-N1-(1-methoxyprop-2-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0114] 6) (E)-N8-hydroxy-N1-(4-methoxybenzyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0115] 7) (E)-N1-(4-fluorophenethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide,

[0116] 8) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(tetrahydrofuran-2-yl)methyl)-2-octenediamide,

[0117] 9) (E)-N1-(2-cyclohexenylethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0118] 10) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(3-(2-oxopyrrolidin-1-yl)propyl)-2-octenediamide,

[0119] 11)(E)-N1-(furan-2-ylmethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0120] 12) (E)-N1-(4-(dimethylamino)benzyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0121] 13) (E)-N8-hydroxy-N1-(2-methoxyethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0122] 14) (E)-N1-cyclohexyl-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0123] 15) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(thiophen-2-ylmethyl)-2-octenediamide,

[0124] 16) (E)-N8-hydroxy-N1-(4-methoxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0125] 17) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(4-(trifluoromethoxy)benzyl)-2-octenediamide,

[0126] 18) (E)-N1-(1-(cyclohexylmethyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0127] 19) (E)-N1-(1-cyclopentylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0128] 20)(E)-N1-(1-benzylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0129] 21)(E)-N8-hydroxy-N1-(1-isopropylpyrrolidin-3-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0130] 22) (E)-N1-(1-(cyclohexanecarbonyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0131] 23) (E)-tert-butyl 3-(8-(hydroxyamino)-2-((naphthalen-1-yloxy)methyl)-8-oxo-2-octenylamido)pyrrolidine-1-carboxylate,

[0132] 24) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(pyrrolidin-3-yl)-2-octenediamide,

[0133] 25) (E)-N1-(1-cyclohexylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-2-yloxy)methyl)-2-octenediamide,

[0134] 26) (E)-N1-(1-cyclopropylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0135] 27) (E)-N1-(1-cyclopropylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0136] 28) (E)-N1-(1-ethylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0137] 29) (E)-N1-(1-ethylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0138] 30) (E)-N8-hydroxy-N1-(2-(1-methylpyrrolidin-2-yl)ethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide,

[0139] 31)(E)-N8-hydroxy-N1-(1-isopropylpiperidin-4-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, and

[0140] 32)(E)-N1-(3-(Dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide.

[0141] According to one embodiment, the HADC inhibitor included in the present invention may include a phosphate salt of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)oct-2-enediamide), whose structural formula is shown in Formula 2.

[0142] [Formula 2]

[0143]

[0144] According to one embodiment, the anti-PD-1 antibody of the present invention may include one or more selected from pembrolizumab, nivolumab, camrelizumab, cemiplizumab, sintilimab, and toripalimab.

[0145] According to one embodiment, the anti-PD-1 antibody of the present invention may be replaced by an anti-PD-L1 antibody. For example, the anti-PD-L1 antibody may include one or more selected from atezolizumab, avelumab, and durvalumab.

[0146] According to another embodiment, the anti-PD-L1 antibody can be replaced by an anti-CTLA4 antibody, an anti-VEGFR antibody, or an anti-VEGF antibody.

[0147] According to one embodiment, the weight ratio of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof to anti-PD-1 antibody or anti-PD-L1 antibody may be 1:0.1 to 1:15, for example 1:1 to 1:5.

[0148] According to one embodiment, the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or its salt may be administered at a dose of 10 to 500 mg / kg, such as 40 to 250 mg / kg, such as 5 to 50 mg / kg.

[0149] According to one embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody can be administered at a dose of 1 to 200 mg / kg, for example, 1 to 10 mg / kg. In another embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody can be administered at a fixed dose of 200 mg, regardless of the weight of the subject.

[0150] According to one embodiment, the target cancer cells of the present invention may be characterized by expression of PD-L1 and may be screened by measuring PD-L1 expression in tumor tissue. For example, cancer cells may be characterized by overexpression or underexpression of PD-L1. Furthermore, for example, cancer cells with a PD-L1 tumor proportion score (TPS) of 1% or higher, such as 20% or higher, or 50% or higher, may be selected as target cancer cells.

[0151] In particular, the cancer cells may include hepatitis A virus-induced liver cancer cells, hepatitis B virus-induced liver cancer cells, hepatitis C virus-induced liver cancer cells, non-virus-related liver cancer cells, metastatic liver cancer cells, colon cancer cells, pancreatic cancer cells, blood cancer cells, melanoma cells, or lung cancer cells. For example, the melanoma may be unresectable or metastatic melanoma, and the lung cancer may be non-small cell lung cancer, which is selected from adenocarcinoma, squamous cell carcinoma, non-squamous cell carcinoma, and large cell carcinoma.

[0152] Additionally, cancer cells may or may not respond to growth regression or death when treated with existing drugs including sorafenib, lenvatinib, regorafenib, nivolumab, or pembrolizumab, and may have been previously treated with these substances.

[0153] According to one embodiment, the target cancer cells of the present invention may or may not have shown progression when previously treated with an anti-CTLA4 antibody or a BRAF inhibitor other than the above substances.

[0154] According to one embodiment, the treatment order of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide or its salt and anti-PD-1 antibody or anti-PD-L1 antibody is not particularly limited and can be any order of simultaneous or sequential administration. Therefore, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or its salt and anti-PD-1 antibody or anti-PD-L1 antibody can be administered simultaneously, sequentially or in reverse order. Specifically, they can be administered simultaneously as a single formulation or as separate formulations, or administered simultaneously, sequentially or in reverse order.

[0155] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof and anti-PD-1 antibody or anti-PD-L1 antibody can be prepared as a single formulation and administered simultaneously.

[0156] Furthermore, the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt, and the anti-PD-1 antibody or anti-PD-L1 antibody can be prepared in separate compositions.

[0157] According to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivatives or salts thereof and anti-PD-1 antibody or anti-PD-L1 antibody can be administered at a dose less than or equal to a therapeutically effective dose. For example, when administered in combination with other types of anti-cancer therapeutic agents, the dose can be adjusted and used as an adjuvant.

[0158] According to one embodiment, when administered in combination with other therapeutic agents, 1) alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or salt thereof is administered before the administration of anti-PD-1 antibody or anti-PD-L1 antibody, or

[0159] 2) Anti-PD-1 antibody or anti-PD-L1 antibody can be administered before administration of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof.

[0160] According to one embodiment, cancer cell death can be induced by treating animals with these two substances for 1 to 40 days, for example, 3 to 21 days. Specifically, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivatives, or salts thereof can be administered repeatedly for 3 weeks. For example, one treatment cycle includes 5 consecutive days of drug administration followed by 2 consecutive days of no administration. In addition, anti-PD-1 antibodies or anti-PD-L1 antibodies can be administered on the first day of the treatment cycle or every other week.

[0161] The method of administration can be selected from a variety of compound administration techniques available in the art, including but not limited to oral, injection, aerosol, parenteral and topical administration. Pharmaceutical compositions can also be obtained by reacting the compound with an inorganic or organic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0162] The treatment method is not particularly limited, but according to one embodiment, alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, its derivative or its phosphate salt can be administered to animals intravenously or orally, and anti-PD1 antibody or anti-PD-L1 antibody can be administered to animals intravenously.

[0163] The pharmaceutical composition of the present invention can be provided in the form of tablets, granules, powders, capsules, dry syrups or injections. Specifically, it can be provided in any convenient form, such as tablets, pills, granules, capsules, suspensions, emulsions or powders suitable for reconstitution with water or other suitable liquid media. In addition, it can be provided in the form of oral dosage forms or injections.

[0164] The pharmaceutical composition for oral administration may comprise one or more diluents selected from microcrystalline cellulose, mannitol and lactose, one or more lubricants selected from talc, magnesium stearate, sodium stearyl fumarate and glyceryl behenate, and one or more binders selected from polyvinyl pyrrolidone, hydroxypropyl methylcellulose and hydroxypropyl cellulose. In addition, for example, the content of the coating layer accounts for 1 to 10 weight percent of 100 weight parts of tablets or capsules. Specifically, for example, the coating layer may comprise a water-soluble coating matrix, and as for the coating matrix, a conventional coating matrix may be used. More specifically, for example, a coating matrix comprising a polyvinyl alcohol derivative, a methacrylic acid derivative and a polyacrylic acid derivative may be included, for example, selected from and hydroxypropyl methylcellulose (HPMC), one or two or more thereof, such as polyvinyl alcohol containing polyvinyl alcohol having relatively excellent moisture and light-blocking effects.

[0165] When preparing granular compositions for oral administration, pure water is preferably not used as a binding solvent due to its unstable moisture stability. However, ethanol, which is easily removed during the manufacturing process, may be used. Magnesium stearate is a conventional lubricant for oral compositions. However, since magnesium stearate may be incompatible with the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide of the present invention, alternatives to magnesium stearate may be used.

[0166] According to one embodiment, in addition to the above-mentioned additives, a pharmaceutically acceptable excipient having excellent compatibility with the compound may be added.

[0167] The injectable composition can be provided in liquid form because the alkylcarbamoylnaphthyloxyoctenoylhydroxyamide is a water-soluble substance with high solubility in water. This composition generally does not require the use of solubilizers and other additives for poorly soluble substances. Minimal additives are preferred because compatibility with additives can be unstable. More specifically, the composition can be prepared by dissolving in nitrogen-purged water for injection followed by freeze-drying.

[0168] According to one embodiment, the method further includes a sterilization step. Sterilization methods may include dry heat sterilization, high pressure or reduced pressure sterilization, filtration sterilization, gas sterilization, radiation sterilization, etc. Filter sterilization may, for example, use a nitrocellulose membrane filter, such as a 0.45 μm filter or a 0.2 μm filter. In the present invention, the method may further include a sterilization step, such as by high temperature reduced pressure sterilization or sterile filtration.

[0169] According to another embodiment of the present invention, a pharmaceutical composition for an anticancer agent may be provided, comprising alkylcarbamoylnaphthyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof, and an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0170] The embodiments of the present invention will be described in detail below so that those skilled in the art can easily implement the present invention. However, the present invention can be implemented in various forms and is not limited to the embodiments described herein.

[0171] Experimental Example 1: Tumor Growth Inhibition Rate in Mouse Models

[0172] As shown in Table 1, in 9 liver cancer mouse models in which Hepa1-6 liver cancer cells were transplanted into mice (C57BL / 6), it was demonstrated that the tumor growth inhibition ability was improved when treated with alkylcarbamoylnaphthyloctenoylhydroxyamide (HDAC inhibitor) or anti-PD-1 antibody, and it was demonstrated that the tumor disappeared when these two substances were used in combination.

[0173] [Table 1]

[0174] Example 1 HDAC inhibitors and anti-PD-1 antibodies Comparative Example 1 HDAC inhibitors Comparative Example 2 Anti-PD-1 antibodies

[0175] (E)-N1-(3-(Dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide phosphate (CG-745, CAS No. 2173017-02-0) was used as the alkylcarbamoylnaphthyloxyoctenoyl hydroxyamide compound, and BP0146 (BioXcell, West Lebanon, NH, USA) was used as the anti-PD-1 antibody.

[0176] 1×10 6 Hepa1-6 liver cancer cells were injected into the left flank of 6-week-old C57BL6 mice. Five days later, they were divided into four groups and administered with vehicle, CG-745 alone, anti-PD-1 antibody alone, or a combination of CG-745 and anti-PD-1 antibody. Tumor volume was defined as (long axis) × (short axis) 2 ×0.5. Mice with tumor volumes less than 1 mm were considered tumor-free by measuring tumor volume and were observed for at least 50 days. Vehicle, CG-745, and anti-PD-1 antibody were administered intraperitoneally (IP). CG-745 was administered at a dose of 20 mg / kg for 5 consecutive days, followed by a 2-day rest (1 treatment cycle), and this treatment cycle was repeated 3 times. On the fifth day of the first treatment cycle of CG-745, anti-PD-1 antibody was administered once together with CG-745 at a dose of 5 mg / kg.

[0177] The average tumor growth inhibition rate of 9 mice was as follows Figure 1The results for each of the 9 mice are shown in Figure 2 shown.

[0178] like Figure 1 As shown, on day 32, the tumor growth inhibition rate for Comparative Example 1 was 73.8%, and the tumor growth inhibition rate for Comparative Example 2 was 84.59%. Furthermore, it was found that after treatment with Comparative Example 1, tumors disappeared in four mice. It was found that, particularly on day 32, the tumor growth inhibition rate for Example 1 was 111.18%, and tumors disappeared in all nine mice. Therefore, it was demonstrated that the anti-PD-1 antibody enhanced the direct or indirect anticancer activity of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide.

[0179] As described above, the present invention demonstrates that the combination of alkylcarbamoylnaphthyloxyoctenoylhydroxyamide as an HDAC inhibitor and an anti-PD-1 antibody has an effective effect in killing cancer cells.

[0180] The above description is merely an illustration of the technical concept of the present invention. A person skilled in the art of the present invention may make various modifications and changes without departing from the essential features of the present invention. In addition, the embodiments disclosed in the present invention are not intended to limit the technical concept of the present invention, but rather to illustrate the technical concept. The scope of the technical concept of the present invention is not limited by these embodiments. The scope of protection of the present invention shall be interpreted by the appended claims, and all technical concepts within the scope of their equivalents shall be interpreted as being included within the scope of the present invention.

Claims

1. Use of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; and an anti-PD-1 antibody in the preparation of a pharmaceutical composition for treating or preventing liver cancer; wherein the pharmaceutical composition has a synergistic effect in inhibiting the growth of cancer cells or killing cancer cells; and the salt is selected from phosphate, tartrate, and a mixture thereof.

2. The use according to claim 1, wherein the weight ratio of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof to the anti-PD-1 antibody is 1:0.1 to 1:

15.

3. The use according to claim 1, wherein the dose of the anti-PD-1 antibody is 1 to 200 mg / kg.

4. The use according to claim 1, wherein the dosage of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof is 10 to 500 mg / kg.

5. The use according to claim 1, wherein the anti-PD-1 antibody comprises one or more compounds selected from the group consisting of pembrolizumab, nivolumab, camrelizumab, cemiplizumab, sintilimab, and toripalimab.

6. The use according to claim 1, wherein the salt of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide is (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide phosphate.

7. The use according to claim 6, wherein the pharmaceutical composition comprises an anti-PD-1 antibody, wherein the anti-PD-1 antibody is BP0146.

8. The use according to any one of claims 1 to 7, wherein: (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; and an anti-PD-1 antibody for preparing a pharmaceutical composition as a single preparation, or for preparing a pharmaceutical composition as two separate preparations.

9. The use according to claim 8, wherein: (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; and an anti-PD-1 antibody for preparing a pharmaceutical composition as a single preparation.

10. The use according to claim 8, wherein: (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; and an anti-PD-1 antibody for preparing a pharmaceutical composition as two separate formulations for simultaneous or sequential administration.

11. The use according to any one of claims 1 to 7, wherein: (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalene-1-yloxy)methyl)-2-octenediamide or its salt; and anti-PD-1 antibody are respectively used to prepare a pharmaceutical composition having a dose less than or equal to a therapeutically effective dose.

12. The use according to any one of claims 1 to 7, wherein: 1) (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof is used for administration before administration of an anti-PD-1 antibody, or 2) The anti-PD-1 antibody is used for administration before administration of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof.

13. The use according to any one of claims 1 to 7, wherein (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof and an anti-PD-1 antibody are used to prepare a pharmaceutical composition as two separate preparations, whereby (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof is for intravenous or oral administration, and the anti-PD-1 antibody is for intravenous administration.

Images