Pharmaceutical Compositions Comprising HDAC Inhibitors and Anti-PD-1 or Anti-PD-L1 Antibodies

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, which can effectively prolong the survival of animals with cancer, tumors, tumor-related disorders, and neoplastic diseases, inhibit proliferative cell growth associated with neoplasia, or cause neoplasia regression.

Description

[Technical field]

[0001] The present invention relates to methods of reducing the growth of and killing cancer cells using a combination of HDAC inhibitors and anti-PD-1 or anti-PD-L1 antibodies.

[0002] This application relates to Patent No. 1978364, registered on May 8, 2019, entitled "Pharmaceutically acceptable salt of alkylcarbamoylnaphthaleneyloxyoctenoyl hydroxyamide or its derivative and preparation method thereof," and claims priority for the use of alkylcarbamoylnaphthaleneyloxyoctenoyl hydroxyamide, its derivative or its salt, which is an epigenome targeting inhibitor, in combination therapy with anti-PD-1 antibody in carcinoma, since the specific immune activity of alkylcarbamoylnaphthaleneyloxyoctenoyl hydroxyamide, its derivative or its salt has been proven to induce an increased anti-cancer effect of anti-PD-1 antibody.

[0003] This application also claims the benefit of priority based on Korean Patent Application No. 2019-0109256, dated September 4, 2019, and all contents disclosed in the documents of said Korean Patent Application are incorporated herein by reference. [Background technology]

[0004] Histones are basic proteins that bind to DNA in the nucleus of eukaryotic cells, and reversible acetylation occurs at the amino group of lysine residues at specific positions in each histone molecule. This histone acetylation reaction forms the higher-order structure of chromatin, regulates the expression of genetic information, and is stably regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). These enzymes are known to regulate gene expression at the transcription level by neutralizing the positive charge of lysine residues (four in the case of H4) at the amino terminus of histones through acetylation to induce transcriptional activity, or by deacetylating and recharging the positive charge to suppress transcription, thereby inducing equilibrium in the acetylation level of histones.

[0005] HDACs have recently been found to be highly expressed under adverse environmental conditions such as hypoxia, low glucose, and cell carcinogenesis, and to promote cancer cell proliferation by inhibiting the expression of tumor suppressor factors, and are recognized as important target factors in regulating cell carcinogenesis and differentiation. In other words, if a highly acetylated state of chromatin inhibits the proliferation and promotes the death of cancer cells, HDACs play a crucial role in inducing cancer cell proliferation through histone deacetylation. This fact is supported by the results that treatment with HDAC inhibitors inhibits the proliferation and angiogenesis of cancer cells.

[0006] On the other hand, PD-1 is known to play an important role in immune regulation and maintenance of peripheral tolerance. PD-1 is expressed at intermediate levels 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] Two known ligands for PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), are expressed in cancers occurring in a variety of tissues. For example, in many cancers, including ovarian, renal, rectal, colon, pancreatic, hepatic, and melanoma, PD-L1 expression correlates with poor prognosis and reduced overall survival, regardless of subsequent treatment.

[0008] Thus, PD-L1-expressing tumor cells are understood to interact with PD-1-expressing T cells to attenuate T cell activation and immune surveillance, thereby contributing to an impaired immune response against tumors.

[0009] It is expected that the efficacy of such antibodies can be further enhanced by administration in conjunction with radiation therapy, surgery, chemotherapy, targeted therapy, signaling pathway inhibitors, immune enhancers, and the like.

[0010] Therefore, there is a need to study a treatment method that combines suitable inhibitors and antibodies as described above for effective application in suppressing the growth and causing necrosis of cancer cells. Summary of the Invention [Problem to be solved by the invention]

[0011] To solve the above problems, the present invention provides 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. [Means for solving the problem]

[0012] The present invention provides a pharmaceutical composition comprising an alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof, and an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0013] According to an embodiment, the weight ratio of the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative, or a salt thereof to the anti-PD-1 antibody or anti-PD-L1 antibody is 1:0.1 to 1:15.

[0014] According to one embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a dose of 1 to 200 mg / kg.

[0015] According to one embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative or its salt is administered at a dose of 10 to 500 mg / kg.

[0016] According to one embodiment, the anti-PD-1 antibody may include one or more selected from the group consisting of pembrolizumab, nivolumab, camrelizumab, cemiplimab, sintilimab, and toripalimab.

[0017] According to one embodiment, the anti-PD-L1 antibody may include one or more selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0018] According to an embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide or a derivative thereof is one or more selected from the group consisting of the following compounds:

[0019] 1) (E)-N1-(3-(1H-imidazol-1-yl)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 2) (E)-N8-hydroxy-N1-(4-hydroxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 3) (E)-N1-(3-(dimethylamino)-2,2-dimethylpropyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 4) (E)-N1-(2-(diisopropylamino)ethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 5) (E)-N8-hydroxy-N1-(1-methoxypropan-2-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 6) (E)-N8-Hydroxy-N1-(4-methoxybenzyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 7) (E)-N1-(4-fluorophenethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 8) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(tetrahydrofuran-2-yl)methyl)-2-octenediamide, 9) (E)-N1-(2-cyclohexenylethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 10) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(3-(2-oxopyrrolidin-1-yl)propyl)-2-octenediamide, 11) (E)-N1-(furan-2-ylmethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 12) (E)-N1-(4-(dimethylamino)benzyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 13) (E)-N8-Hydroxy-N1-(2-methoxyethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 14) (E)-N1-Cyclohexyl-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 15) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(thiophen-2-ylmethyl)-2-octenediamide, 16) (E)-N8-Hydroxy-N1-(4-methoxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 17) (E)-N8-Hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(4-(trifluoromethoxy)benzyl)-2-octenediamide, 18) (E)-N1-(1-(cyclohexylmethyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 19) (E)-N1-(1-cyclopentylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 20) (E)-N1-(1-benzylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 21) (E)-N8-Hydroxy-N1-(1-isopropylpyrrolidin-3-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 22) (E)-N1-(1-(cyclohexanecarbonyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 23) (E)-3-(8-(hydroxyamino)-2-((naphthalen-1-yloxy)methyl)-8-oxo-2-octenamido)pyrrolidine-1-carboxylic acid t-butyl ester, 24) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(pyrrolidin-3-yl)-2-octenediamide, 25) (E)-N1-(1-cyclohexylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-2-yloxy)methyl)-2-octenediamide, 26) (E)-N1-(1-cyclopropylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 27) (E)-N1-(1-cyclopropylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 28) (E)-N1-(1-ethylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 29) (E)-N1-(1-ethylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 30) (E)-N8-hydroxy-N1-(2-(1-methylpyrrolidin-2-yl)ethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 31) (E)-N8-Hydroxy-N1-(1-isopropylpiperidin-4-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, and 32) (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide.

[0020] According to one embodiment, the salt of alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide is (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalene-1-yloxy)methyl)-2-octenediamide phosphate.

[0021] According to one embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative or a salt thereof, and the anti-PD-1 antibody or anti-PD-L1 antibody are administered simultaneously in one formulation or simultaneously, sequentially or in reverse order in separate formulations.

[0022] According to one embodiment, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, derivative or salt thereof, and the anti-PD-1 antibody or anti-PD-L1 antibody are present as a single formulation.

[0023] According to an embodiment, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, a derivative thereof or a salt thereof, and the anti-PD-1 antibody or anti-PD-L1 antibody are each present as separate compositions.

[0024] According to one embodiment, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative or a salt thereof, and the anti-PD-1 antibody or anti-PD-L1 antibody are each administered in a sub-therapeutic dose.

[0025] According to one embodiment, 1) the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative, or a salt thereof is administered prior to administration of the anti-PD-1 antibody or anti-PD-L1 antibody, or 2) the anti-PD-1 antibody or anti-PD-L1 antibody is administered prior to administration of the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative, or a salt thereof.

[0026] According to one embodiment, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, derivative, or salt thereof is administered intravenously or orally to the animal, and the anti-PD-1 antibody or anti-PD-L1 antibody is formulated as a separate agent to be administered by intravenous infusion to the animal.

[0027] According to one embodiment, the compositions are synergistic in inhibiting the growth or killing of cancer cells.

[0028] According to one embodiment, the cancer cells are characterized by expression of PD-L1, and the cancer cells are hepatoma virus A-related hepatoma cells, hepatoma virus B-related hepatoma cells, hepatoma virus C-related hepatoma cells, non-virus-related hepatoma cells, metastatic hepatoma cells, colorectal cancer cells, pancreatic cancer cells, blood cancer cells, melanoma cells or lung cancer cells.

[0029] According to one embodiment, the cancer cells are selected by a method comprising measuring the expression of PD-L1 in tumor tissue.

[0030] According to one embodiment, the tumor proportion score (TPS) for PD-L1 expression in the cancer cells is 1% or more.

[0031] Further details of the embodiment of the present invention are included in the following detailed description. Effect of the Invention

[0032] 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 of the present invention can effectively kill cancer cells.

[0033] Cancers, tumors, tumor-related disorders and neoplastic diseases are mostly life-threatening conditions characterized by rapid cell proliferation and growth. The methods of the present invention can effectively affect the prolongation of survival of animals with said diseases or disorders, the inhibition of proliferative cell growth associated with neoplasms or the regression of neoplasms. [Brief description of the drawings]

[0034] [Figure 1] 1 is a graph showing tumor growth inhibition rates in mouse models. [Diagram 2] 1 is a graph showing tumor growth inhibition rates in mouse models. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0035] Since the present invention can have various embodiments with various modifications and variations, specific embodiments are illustrated in the drawings and described in detail. However, this is not intended to limit the present invention to a specific embodiment, and it should be understood that the present invention includes all modifications, equivalents, or alternatives within the spirit and technical scope of the present invention. In describing the present invention, if it is determined that a detailed description of related publicly known technology may obscure the gist of the present invention, the detailed description will be omitted.

[0036] Hereinafter, 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 embodiments of the present invention will be described in more detail.

[0037] The term "abnormal cell growth" as used herein refers to cell growth independent of normal regulatory mechanisms, including abnormal growth of normal cells and growth of abnormal cells, e.g., loss of contact inhibition, i.e., loss of mechanisms that inhibit growth and division when cells grow and come into contact with surrounding cells or tissues.

[0038] As used herein, the term "neoplasia" is a non-normal, unregulated, unorganized proliferation of cells that is distinguished from normal cells by autonomous growth and somatic mutation. As neoplastic cells grow and divide, they transmit their genetic mutations and proliferative characteristics to progeny cells. A neoplasm, or tumor, is the accumulation of neoplastic cells. In some embodiments, neoplasia is benign or malignant.

[0039] As used herein, the term "metastasis" refers to the dissemination of tumor cells through lymphatic or blood vessels. Metastasis also refers to the migration of tumor cells by direct extension through serous cavities, or subarachnoid or other spaces. The migration of tumor cells to other sites in the body through the process of metastasis allows the establishment of a neoplasm at a site distant from the site of initial appearance.

[0040] As used herein, the term "subject" refers to an animal, including, but not limited to, a primate (e.g., a human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject" and "patient" are used interchangeably herein, e.g., in reference to a mammalian subject, e.g., a human subject.

[0041] As used herein, the terms "treat" and "treatment" are meant to include alleviating or preventing a disorder, disease, or condition, or one or more symptoms associated with a disorder, disease, or condition, or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself.

[0042] As used herein, the term "therapeutically effective amount" refers to an amount of a compound that, upon administration, is sufficient to prevent the onset of, or alleviate to some extent, one or more of the symptoms of, the disorder, disease, or condition being treated. The term "therapeutically effective amount" also refers to an amount of a compound sufficient to elicit the biological or medical response in a cell, tissue, system, animal, or human that is desired by a researcher, veterinarian, physician, or clinician.

[0043] The term "pharmaceutical composition" as used herein is interchangeable with "pharmaceutical composition" and "pharmaceutical acceptable composition" and refers to a formulation of any organic or inorganic compound that is relatively non-toxic and has an effective effect that is harmless to the subject to which it is administered, and that does not reduce the efficacy of the drug due to side effects resulting from the composition, does not induce significant irritation in the subject to which the compound is administered, and does not impair the biological activity and physical properties of the compound.

[0044] As used herein, the term "pharmaceutically acceptable carrier", "pharmaceutically acceptable excipient", "physiologically acceptable carrier" or "physiologically acceptable excipient" means a pharmaceutically acceptable substance, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each component must be "pharmaceutically acceptable" in terms of compatibility with the other components of a pharmaceutical formulation. It must also be suitable for use in contact with human and animal tissues or organs without undue toxicity, irritation, allergic response, immunogenicity or other problems or complications, commensurate with a reasonable benefit / risk ratio (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).

[0045] The term "pharmaceutical composition" as used herein means a mixture of the compounds disclosed herein and other chemical components, such as pharma- ceutical acceptable diluents, carriers, etc. The compounds can be easily administered to an organism through the pharmaceutical composition.

[0046] In the present invention, each component drug of the pharmaceutical composition may be a separate formulation or a single formulation, and the component drugs may be administered simultaneously, sequentially or in reverse order. In addition, in the present invention, the pharmacologic effective amount, administration time, administration interval, administration route, treatment period, etc. of each component drug of the pharmaceutical composition may be the same or different from each other.

[0047] The term "subject" as used herein is interchangeable with "subject individual" and "subject organism" and refers to any animal, including humans, that has been or may be induced to become infected with a bacterium or a resistant strain.

[0048] The term "death" as used in the present invention includes both "cell necrosis" and "cell death".

[0049] As discussed herein, "angiogenesis" is important in tumor formation and metastasis. Angiogenic factors have been found to be associated with several solid tumors, such as rhabdomyosarcoma, retinoblastoma, Ewing sarcoma, neuroblastoma, and osteosarcoma. Tumors cannot expand without a blood supply to provide nutrients and cellular waste products. Tumors in which angiogenesis is important include solid tumors, such as renal cell carcinoma, hepatocellular carcinoma, and benign tumors, such as acoustic neuroma and neurofibromatosis. Angiogenesis is associated with blood-borne tumors, such as leukemia. Angiogenesis is believed to play a key role in bone marrow abnormalities that lead to leukemia. Preventing angiogenesis can halt the growth of cancerous tumors and the resulting damage to the subject caused by the tumor's presence.

[0050] The present invention provides pharmaceutical compositions and methods for inhibiting the growth of or killing cancer cells using a combination of an HDAC inhibitor and an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0051] Histone deacetylases (HDACs) are a family of at least 18 enzymes that are divided into three classes (classes I, II, and III). Class I HDACs include, but are not limited to, HDACs 1, 2, 3, and 8. Class I HDACs are found in the nucleus and are considered to be associated with transcriptional repressors. Class II HDACs include, but are not limited to, HDACs 4, 5, 6, 7, and 9, and are found in the nucleus as well as the cytoplasm. Class III HDACs are considered to be NAD-dependent proteins and include, but are not limited to, members of the sirtuin protein family. Non-limiting examples of sirtuin proteins include SIRT1-7. As used herein, the term "selective HDAC" refers to an HDAC inhibitor that does not interact with any of the three HDAC classes.

[0052] 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), cyclic tetrapeptides containing 2-amino-8-oxo-9,10-epoxy-decanoic acid (i.e., trapoxin A), and cyclic peptides that do not contain the 2-amino-8-oxo-9,10-epoxy-decanoic acid moiety (i.e., FK228).

[0053] HDAC inhibitors are broadly classified into general HDAC inhibitors and selective HDAC inhibitors. Although there is a wide variety of structural diversity among known HDAC inhibitors, they share common features: a moiety that interacts with the enzyme active site, and a side chain that is placed inside the channel to guide the active site. This is seen in hydroxamates, such as SAHA (suberoylanilide hydroxamic acid), where the hydroxamate group is considered to interact with the active site. In the case of depsipeptides, a free thiol group attached to a 4-carbon alkenyl chain, which interacts with the active site, is considered to be formed through intracellular reduction of a disulfide bond.

[0054] The difference between HDAC inhibitors lies in the way they interact with the periphery of the HDAC channel, which is at the opposite end of the channel to the active site. Such interactions between HDAC inhibitors and the periphery of the channel are believed to be responsible, at least in part, for the differences in HDAC selectivity observed in part between pan HDAC inhibitors such as SAHA and selective HDAC inhibitors such as depsipeptide.

[0055] Alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamides are HDAC inhibitors currently in clinical investigation in multiple solid tumor and hematological cancer types. Alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamides are rapidly absorbed, with half-lives of approximately 7-8 hours upon intravenous administration. Importantly, histone acetylation changes persist for several days after administration.

[0056] On the other hand, programmed cell death-1, or PD-1, is a cell surface receptor that is 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, and the full-length PD-1 cDNA encodes a protein containing 288 amino acid residues with 60% homology to mouse PD-1. It is present on CD4-CD8- (double negative) thymocytes during thymic development and is expressed upon activation in mature hematopoietic cells, e.g., T and B cells, NKT cells, and monocytes, after prolonged antigen exposure. PD-L1 has been shown to be expressed on a large number of mouse and human tumors (mostly inducible by IFN-gamma on PD-L1-negative tumor cell lines) and has been hypothesized 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 numerous primary tumor biopsies from head and neck tumors as well as liver, lung, ovarian, cervical, skin, colon, neuroblastoma, bladder, breast, kidney, esophagus, stomach, oral squamous cell carcinoma, urothelial cell carcinoma, and pancreatic cancers (see references: [Brown JA et al., J. Immunol. 170:1257-1266(2003)]; [Dong H. et al., Nat. Med. 8:793-800(2002)]; [Wintterle et al., Cancer Res. 63:7462-7467(2003)]; [Strome SE et al., Cancer Res., 63:6501-6505(2003)]; [Thompson RH et al., Cancer Res. 1999, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2030, 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, 2040, 2041, 2042, 2043, 2044, 2045, Res.66:3381-5(2006)];[Thompson et al.,Clin.Cancer Res.13:1757-61(2007)];[Nomi T.et al.,Clin.Cancer Res.13:2151-7.(2007)]).Expression of PD-L1 ligands on tumor cells correlates with poor prognosis in cancer patients across multiple tumor types (reviewed in Okazaki and Honjo, Int. Immunol. 19:813-824 (2007)).

[0057] It is also known that high expression of PD-L1 on tumor cells correlates with poor prognosis and survival in various other solid tumor types. The PD-1 / PD-L1 pathway is believed to play an important role in tumor immune evasion, and it is understood and considered to be an attractive target for therapeutic intervention in several solid organ types. Several PD-1 and PD-L1 antibodies are in clinical development, and generally most have been reported to be well tolerated, although dose-limiting toxicity was not reached in their Phase I trials. Through various studies, it has been revealed that the interaction of PD-1 with its ligands (PD-L1 and PD-L2) suppresses lymphocyte proliferation in vitro and in vivo. Thus, it is believed that the binding of the ligand PD-L1 to PD-1 downregulates effector antitumor T cell activity and promotes immune evasion.

[0058] Disruption of PD-1 / PD-L1 interaction has also been shown to increase T cell proliferation and cytokine production, and block cell cycle progression. In vitro studies of PD-1 blockade with PD-1-specific antibodies have shown that it enhanced cytotoxic T cell responses to liver cancer-specific antigens, including an increase in the frequency of IFN-γ-secreting antigen-specific cells. Therefore, targeting PD-1 may serve as an effective therapeutic strategy for cancer.

[0059] The primary method of targeting PD-1 is through the development of genetically engineered monoclonal antibodies that inhibit function by interfering with the binding of PD-1 to PD-L1. Anti-PD-1 antibodies and 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 an alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, a derivative or salt thereof, and an anti-PD-1 antibody or antigen-binding portion thereof, wherein the antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof. In certain embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is a humanized antibody. In some embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is a human antibody. In some embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is a monoclonal antibody or antigen-binding portion thereof.

[0060] According to one embodiment, the HDAC inhibitor may include an alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof. The alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide is represented by the structural formula 1.

[0061] [ka] R1 is a C substituted or unsubstituted 1-3 Alkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyl C 1-3 Alkyl, benzyl, C 1-3 Alkyl or C 3-8 Cycloalkylcarbonyl-substituted or unsubstituted pyrrolidine, C 1-3 Alkyl or C 3-8 Cycloalkyl-substituted piperidine, furan, or C 3-8 is cycloalkyl, However, unsubstituted C 1-2 Alkyl and C1-2 Alkylpyrrolidinyl substituted C 1-2 Except for alkyl, The salts may be selected from phosphoric acid salts, tartaric acid salts, stearic acid salts, gluconic acid salts, fumaric acid salts, naphthoic acid salts, 1-hydroxy-2 salts, and mixtures thereof, for example, phosphate salts.

[0062] According to an embodiment, the salt may be selected from phosphates, tartrates, and mixtures thereof, which have relatively high stability and water solubility when stored at room temperature, and may include, for example, phosphates.

[0063] The alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide of the above structural formula 1 or a derivative thereof may be selected from the group consisting of the following compounds.

[0064] 1) (E)-N1-(3-(1H-imidazol-1-yl)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 2) (E)-N8-hydroxy-N1-(4-hydroxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 3) (E)-N1-(3-(dimethylamino)-2,2-dimethylpropyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 4) (E)-N1-(2-(diisopropylamino)ethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)octenediamide, 5) (E)-N8-hydroxy-N1-(1-methoxypropan-2-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 6) (E)-N8-Hydroxy-N1-(4-methoxybenzyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 7) (E)-N1-(4-fluorophenethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 8) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(tetrahydrofuran-2-yl)methyl)-2-octenediamide, 9) (E)-N1-(2-cyclohexenylethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 10) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(3-(2-oxopyrrolidin-1-yl)propyl)-2-octenediamide, 11) (E)-N1-(furan-2-ylmethyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 12) (E)-N1-(4-(dimethylamino)benzyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 13) (E)-N8-Hydroxy-N1-(2-methoxyethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 14) (E)-N1-Cyclohexyl-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 15) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(thiophen-2-ylmethyl)-2-octenediamide, 16) (E)-N8-Hydroxy-N1-(4-methoxyphenethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 17) (E)-N8-Hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(4-(trifluoromethoxy)benzyl)-2-octenediamide, 18) (E)-N1-(1-(cyclohexylmethyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 19) (E)-N1-(1-cyclopentylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 20) (E)-N1-(1-benzylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 21) (E)-N8-Hydroxy-N1-(1-isopropylpyrrolidin-3-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 22) (E)-N1-(1-(cyclohexanecarbonyl)pyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 23) (E)-3-(8-(hydroxyamino)-2-((naphthalen-1-yloxy)methyl)-8-oxo-2-octenamido)pyrrolidine-1-carboxylic acid t-butyl ester, 24) (E)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-N1-(pyrrolidin-3-yl)-2-octenediamide, 25) (E)-N1-(1-cyclohexylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-2-yloxy)methyl)-2-octenediamide, 26) (E)-N1-(1-cyclopropylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 27) (E)-N1-(1-cyclopropylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 28) (E)-N1-(1-ethylpiperidin-4-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 29) (E)-N1-(1-ethylpyrrolidin-3-yl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 30) (E)-N8-hydroxy-N1-(2-(1-methylpyrrolidin-2-yl)ethyl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, 31) (E)-N8-Hydroxy-N1-(1-isopropylpiperidin-4-yl)-2-((naphthalen-1-yloxy)methyl)-2-octenediamide, and 32) (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide.

[0065] According to one embodiment, the HDAC inhibitor included in the present invention includes a phosphate salt of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)oct-2-enediamide, the structural formula of which is as shown in structural formula 2.

[0066] [ka]

[0067] According to one embodiment, the anti-PD-1 antibody of the present invention may include one or more selected from the group consisting of pembrolizumab, nivolumab, camrelizumab, cemiplimab, sintilimab, and toripalimab.

[0068] According to an embodiment, the anti-PD-1 antibody of the present invention may be substituted for an anti-PD-L1 antibody, for example, the anti-PD-L1 antibody may include one or more selected from the group consisting of atezolizumab, avelumab, durvalumab, etc.

[0069] According to other embodiments, the anti-PD-1 antibody may be replaced by an anti-CTLA4 antibody, an anti-VEGFR antibody, or an anti-VEGF antibody.

[0070] According to an embodiment, the weight ratio of the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative, or a salt thereof to the anti-PD-1 antibody or anti-PD-L1 antibody is 1:0.1 to 1:15, for example, 1:1 to 1:5.

[0071] According to an embodiment, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative or its salt is used in an amount of 10 to 500 mg / kg, for example, 40 to 250 mg / kg, for example, 5 to 50 mg / kg.

[0072] According to one embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody is used in an amount of 1-200 mg / kg, for example, 1-10 mg / kg, and in another embodiment, the anti-PD-1 antibody or anti-PD-L1 antibody is used in a fixed dose of 200 mg, regardless of the weight of the subject.

[0073] According to an embodiment, the cancer cells to which the present invention is applied are characterized by expression of PD-L1, and the cancer cells can be selected by measuring the expression of PD-L1 in tumor tissue. For example, the cancer cells are characterized by overexpression or underexpression of PD-L1. For example, the tumor proportion score (TPS) of the PD-L1 expression of the cancer cells can be selected as the cancer cells to which the present invention is applied, for example, 1% or more, e.g., 20% or more, 50% or more.

[0074] The cancer cells may specifically include, for example, hepatoma virus A hepatoma cells, hepatoma virus B hepatoma cells, hepatoma virus C hepatoma cells, non-virus-related hepatoma cells, metastatic hepatoma cells, colon cancer cells, pancreatic cancer cells, blood cancer cells, melanoma cells, or lung cancer cells. For example, the melanoma is unresectable or metastatic melanoma, and for example, the lung cancer is non-small cell lung cancer, and the non-small cell lung cancer may be selected from adenocarcinoma, squamous cell carcinoma, non-squamous cell carcinoma, and large cell carcinoma.

[0075] In addition, the cancer cells may or may not respond to reduced growth or death when treated with existing drugs, including sorafenib, lenvatinib, regorafenib, nivolumab, or pembrolizumab, and have been treated with a prior therapy using such agents.

[0076] According to one embodiment, the cancer cells to which the present invention is applicable are those that have shown or have not shown progress through prior therapy using an anti-CTLA4 antibody or a BRAF inhibitor, in addition to the above types.

[0077] According to an embodiment, the order of administration of the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide or its salt and the anti-PD-1 antibody or anti-PD-L1 antibody is not particularly limited and may be administered in any order, simultaneously or sequentially. That is, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative or its salt and the anti-PD-1 antibody or anti-PD-L1 antibody are administered simultaneously, sequentially or in reverse order, specifically, simultaneously as one formulation, or simultaneously, sequentially or in reverse order as separate formulations.

[0078] According to one embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, derivative or salt thereof and the anti-PD-1 antibody or anti-PD-L1 antibody are present in a single formulation and are administered simultaneously.

[0079] In addition, the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative or its salt, and the anti-PD-1 antibody or anti-PD-L1 antibody each exist as an independent composition.

[0080] According to an embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative or its salt, and the anti-PD-1 antibody or anti-PD-L1 antibody are each administered at a sub-therapeutic dose, for example, when administered in combination with another type of anti-cancer therapeutic agent, the dose may be adjusted to be supplementary.

[0081] According to one embodiment, when used in combination with other therapeutic agents, the order of administration is 1) the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative, or a salt thereof is administered before the anti-PD-1 antibody or anti-PD-L1 antibody, or 2) the anti-PD-1 antibody or anti-PD-L1 antibody is administered before the alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide, its derivative, or a salt thereof.

[0082] According to one embodiment, the two substances are administered to an animal for 1-40 days, for example, 3-21 days, to induce the death of cancer cells. Specifically, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative or its salt is administered for 3 weeks, for example, with 5 consecutive days and 2 days off as one cycle. The anti-PD-1 antibody or anti-PD-L1 antibody is administered on the first day of the treatment cycle, and then every other week.

[0083] The method of administration can be selected from a number of compound administration techniques existing 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, etc.

[0084] The treatment method is not particularly limited, but according to one embodiment, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, its derivative, or its phosphate is administered intravenously or orally to the animal, and the anti-PD-1 antibody or anti-PD-L1 antibody is administered to the animal by intravenous infusion.

[0085] 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, pellets, granules, capsules, suspensions, emulsions or powders suitable for reconstitution with water or other suitable liquid medium.It can also be applied in the form of oral administration or injections.

[0086] The pharmaceutical composition for oral administration may include one or more diluents selected from the group consisting of microcrystalline cellulose, mannitol, lactose, and lactose, one or more lubricants selected from the group consisting of talc, magnesium stearate, sodium stearyl fumarate, and glyceryl behenate, and one or more binders selected from the group consisting of polyvinylpyrrolidone, hydroxypropylmethylcellulose, and hydroxypropylcellulose. In addition, the composition may include a coating layer in an amount of 1 to 10% by weight based on 100 parts by weight of the tablet or capsule. Specifically, the coating layer may include a water-soluble coating base, and a commonly used coating base may be used. More specifically, the coating layer may include a coating base containing a polyvinyl alcohol derivative, a methacrylic acid derivative, and a polyacrylic acid derivative, and may include one or more selected from the group consisting of Opadry®, Kollicoat®, and hydroxypropylmethylcellulose (HPMC), and may include polyvinyl alcohol-containing Opadry®, which has a relatively excellent moisture and light blocking effect.

[0087] When preparing granules of a composition for oral administration, it is preferable not to use purified water as a binding solvent, taking into consideration its unstable water stability, and ethanol, which is easy to remove during the manufacturing process, can be used. Magnesium stearate, a lubricant used in conventional compositions for oral administration, has poor compatibility with the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide of the present invention, so a substitute can be used.

[0088] According to an embodiment, in addition to the above additives, pharmaceutical excipients having excellent compatibility with the compounds may be added.

[0089] As an injectable composition, the alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide component is provided in the form of a liquid phase since it is a water-soluble substance that dissolves well in water, and it is not essential to use solubilizers and other additives that are generally used for poorly soluble substances, and since compatibility with additives is unstable, it is preferable to use a minimum amount of additives. More specifically, it can be prepared by dissolving in nitrogen-purged water for injection and then lyophilizing.

[0090] According to an embodiment, a sterilization step may be further included. Sterilization methods may include dry heat sterilization, pressure or vacuum sterilization, filtration sterilization, gas sterilization, radiation sterilization, etc. For filtration sterilization, for example, a nitrocellulose membrane filter may be used, and filters such as 0.45 μm and 0.2 μm may be used. In the present invention, for example, a sterilization step may be further included by high-temperature vacuum sterilization or aseptic filtration.

[0091] According to another embodiment of the present invention, there is provided a pharmaceutical composition for use as an anti-cancer agent, comprising an alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide, a derivative thereof, or a salt thereof, and an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0092] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention may be embodied in various different forms and should not be construed as limited to the embodiments set forth herein. EXAMPLES

[0093] Experimental example 1: Tumor growth inhibition rate in mouse models Nine mice (C57BL / 6) were transplanted with liver cancer cell line Hepa1-6 to generate liver cancer model. As shown in Table 1, treatment with alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide (HDAC inhibitor) or anti-PD-1 antibody improved tumor growth inhibition, and administration of the two substances in combination caused tumor disappearance.

[0094] [Table 1]

[0095] The alkylcarbamoylnaphthalenyloxyoctenoylhydroxyamide compound used was (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide phosphate (CG-745, CAS No. 2173017-02-0), and the anti-PD-1 antibody used was BP0146 (BioXcell, West Lebanon, NH, USA).

[0096] 1 x 10 6 Hepa1-6 hepatoma cells were implanted into the left flank. Five days later, the mice were divided into four groups for vehicle, CG-745 alone, anti-PD-1 antibody alone, or CG-745 and anti-PD-1 antibody combined administration. Tumor volumes were measured using a longitudinal x transverse axis (LSA) scale. 2The tumor volume was calculated as 0.5 × 0.5. Mice with tumors smaller than 1 mm at the time of tumor volume measurement were considered to have disappeared tumors 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 period, repeated three times. Anti-PD-1 antibody was administered once together with CG-745 at a dose of 5 mg / kg on the 5th day of the first CG-745 cycle.

[0097] The average tumor growth inhibition rate for the nine mice is shown in FIG. 1, and the results for each of the nine mice are shown in FIG.

[0098] As shown in the graph of FIG. 1, it was confirmed that Comparative Example 1 and Comparative Example 2 showed tumor growth inhibition rates of 73.8% and 84.59%, respectively, on the 32nd day. In addition, it was confirmed that tumors disappeared in 4 mice treated with Comparative Example 1. In particular, a tumor growth inhibition rate of 111.18% was confirmed on the 32nd day in Example 1, and it was confirmed that tumors disappeared in all 9 mice. Therefore, it was confirmed that the direct and indirect anti-cancer effects of alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide were enhanced by anti-PD-1 antibody.

[0099] As described above, the present invention confirmed that the combination of an alkylcarbamoylnaphthaleneyloxyoctenoylhydroxyamide as an HDAC inhibitor and an anti-PD-1 antibody has an effective effect on killing cancer cells.

[0100] The above description is merely an illustrative example of the technical idea of ​​the present invention, and those skilled in the art can make various modifications and variations without departing from the essential characteristics of the present invention. Furthermore, the embodiments disclosed in the present invention are for illustrative purposes only, and are not intended to limit the technical idea of ​​the present invention. The scope of the present invention should be interpreted according to the following claims, and any technical idea within the scope equivalent thereto should be interpreted as being included in the scope of the present invention.

Claims

1. (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof, A pharmaceutical composition for treating or preventing cancer, comprising an anti-PD-1 antibody or an anti-PD-L1 antibody.

2. 2. The pharmaceutical composition according to claim 1, wherein the weight ratio of the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof to the anti-PD-1 antibody or anti-PD-L1 antibody is 1:0.1 to 1:

15.

3. The pharmaceutical composition according to claim 1 or 2, wherein the dose of the anti-PD-1 antibody or anti-PD-L1 antibody is 1 to 200 mg / kg.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the dose of the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or its salt is 10 to 500 mg / kg.

5. The pharmaceutical composition of any one of claims 1 to 4, wherein the anti-PD-1 antibody comprises one or more compounds selected from the group consisting of pembrolizumab, nivolumab, camrelizumab, cemiplimab, sintilimab, and toripalimab.

6. The pharmaceutical composition of any one of claims 1 to 5, wherein the anti-PD-L1 antibody comprises one or more compounds selected from the group consisting of atezolizumab, avelumab, and durvalumab.

7. 2. The pharmaceutical composition of claim 1, wherein the pharma- ceutically acceptable salt of (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide is a phosphate salt.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition comprises an anti-PD-1 antibody, wherein the anti-PD-1 antibody is BP0146.

9. the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; The pharmaceutical composition according to any one of claims 1 to 8, wherein the anti-PD-1 antibody or the anti-PD-L1 antibody is present as a single formulation.

10. the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; The pharmaceutical composition according to any one of claims 1 to 8, wherein the anti-PD-1 antibody or the anti-PD-L1 antibody is prepared as two separate formulations for simultaneous or sequential administration.

11. the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof; The pharmaceutical composition according to any one of claims 1 to 10, wherein the anti-PD-1 antibody or the anti-PD-L1 antibody is administered at a dose equal to or less than a therapeutically effective dose in monotherapy.

12. 1) the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof is for administration prior to administration of the anti-PD-1 antibody or anti-PD-L1 antibody; or 2) The pharmaceutical composition according to any one of claims 1 to 8, wherein the anti-PD-1 antibody or anti-PD-L1 antibody is for administration prior to administration of the (E)-N1-(3-(dimethylamino)propyl)-N8-hydroxy-2-((naphthalen-1-yloxy)methyl)-2-octenediamide or a salt thereof.

13. The pharmaceutical composition according to any one of claims 1 to 8, wherein the (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 or anti-PD-L1 antibody is for intravenous administration.

14. The pharmaceutical composition according to any one of claims 1 to 13, which has a synergistic effect on inhibiting the growth of and killing cancer cells.

15. The pharmaceutical composition of claim 14, wherein the cancer cells are characterized by expression of PD-L1.

16. The pharmaceutical composition according to claim 14 or 15, wherein the cancer cells are hepatoma cells caused by hepatoma virus A, hepatoma cells caused by hepatoma virus B, hepatoma cells caused by hepatoma virus C, non-virus-related hepatoma cells, metastatic hepatoma cells, colon cancer cells, pancreatic cancer cells, blood cancer cells, melanoma cells or lung cancer cells.

17. The pharmaceutical composition according to any one of claims 14 to 16, wherein the cancer cells are selected by a method comprising a step of measuring expression of PD-L1 in tumor tissue.

18. The pharmaceutical composition according to any one of claims 14 to 17, wherein the tumor proportion score (TPS) for PD-L1 expression in the cancer cells is 1% or more.

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