Composition for enhancing anticancer effect of sorafenib, comprising sash1 overexpression vector as active ingredient

Abstract

The present invention relates to a composition for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector as an active ingredient. The composition comprises a polynucleotide sequence for inducing the overexpression of SASH1 (SAM and SH3 domain-containing protein 1) comprising a sequence commonly expressed by isoforms of a SASH1 gene. Since the vector of the present invention has an excellent anticancer effect by itself while increasing the anticancer effect of sorafenib, the composition comprising the SASH1 overexpression vector and / or sorafenib as an active ingredient can be usefully used as a substance for preventing or treating cancer.

Description

Composition for enhancing the anticancer effect of sorafenib containing a SASH1 overexpression vector as an active ingredient

[0001] The present invention relates to a composition for enhancing the anticancer effect of sorafenib, which comprises a SASH1 overexpression vector as an active ingredient.

[0002] This invention claims priority to Republic of Korea Patent Application No. 10-2023-0168172, filed on November 28, 2023, the entire disclosure of which is incorporated herein by reference.

[0003]

[0004] Surgery, radiation therapy, and chemotherapy are used to treat cancer. Today, there are approximately 60 different anticancer drugs available, and with active research on new anticancer drugs, more and more effective drugs are being developed. However, these drugs have a fatal drawback: when administered repeatedly and over long periods of time or when the cancer relapses, cancer cells develop resistance to them, rendering them ineffective.

[0005] Sorafenib (Sorafenib, Nexavar) @ ) is an oral multi-kinase inhibitor that targets tyrosine kinase receptors and exerts antiangiogenic and antiproliferative effects. According to new guidelines from the Asia-Pacific Association for the Study of the Liver (APASL) and the European Association for the Study of the Liver (EASL), sorafenib is recommended as a first-line treatment for patients with advanced hepatocellular carcinoma who are unsuitable for locoregional therapy. The American Association for the Study of Liver Diseases (AASD) also recommends sorafenib as a systemic agent.

[0006] However, sorafenib exhibits short-term anticancer effects due to a rapid decrease in drug sensitivity and the acquisition of tumor cell resistance. Typically, patients with hepatocellular carcinoma receiving sorafenib develop resistance within six months, with an overall survival increase of only three months. Several mechanisms, including the tumor microenvironment, epithelial-to-mesenchymal transition, and cancer stem cells, are known to be associated with low tumor cell sensitivity to sorafenib.

[0007] Therefore, research on substances and combinations to overcome sorafenib resistance and enhance anticancer effects is actively underway, but few reports have been published to date.

[0008]

[0009] One object of the present invention is to provide a composition for enhancing the anticancer effect of sorafenib, which comprises as an active ingredient a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression.

[0010] Another object of the present invention is to provide a composition for co-administration with sorafenib, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0011] Another object of the present invention is to provide an anticancer adjuvant pharmaceutical composition comprising, as an active ingredient, a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression.

[0012] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0013] Another object of the present invention is to provide a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a pharmaceutical composition for preventing or treating cancer comprising sorafenib as an active ingredient.

[0014] Another object of the present invention is to provide a kit for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

[0015] Another object of the present invention is to provide a kit for preventing or treating cancer, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and instructions.

[0016]

[0017] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0018]

[0019] The present invention provides a composition for enhancing the anticancer effect of sorafenib, comprising as an active ingredient a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression.

[0020] In one embodiment of the present invention, the polynucleotide sequence for inducing SASH1 overexpression may include, but is not limited to, a sequence commonly expressed in a SASH1 gene isoform.

[0021] In one embodiment of the present invention, the commonly expressed sequence may be commonly expressed in exon 20 of the SASH1 gene homolog, but is not limited thereto.

[0022] In one embodiment of the present invention, the commonly expressed sequence may include, but is not limited to, a portion of exon 20 of the SASH1 gene.

[0023] In one embodiment of the present invention, the commonly expressed sequence may include, but is not limited to, a polynucleotide sequence of 500 bp to 3,750 bp in length of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0024] In one embodiment of the present invention, exon 20 may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 4.

[0025] In one embodiment of the present invention, exon 20 may include, but is not limited to, the 4,199th to 7,949th bases from the 5′ end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0026] In one embodiment of the present invention, a portion of the exon 20 sequence may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 1.

[0027] In one embodiment of the present invention, a portion of the sequence of exon 20 may include, but is not limited to, bases 6,608 to 7,108 from the 5′ end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0028] In one embodiment of the present invention, the vector may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 1.

[0029] In one embodiment of the present invention, the vector may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 2.

[0030] In one embodiment of the present invention, the vector may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 3.

[0031] In one embodiment of the present invention, the cancer may be at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC), but is not limited thereto.

[0032] The present invention provides a composition for co-administration with sorafenib, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0033] The present invention provides an anticancer adjuvant pharmaceutical composition comprising, as an active ingredient, a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression.

[0034] The present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0035] The present invention provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a pharmaceutical composition for preventing or treating cancer comprising sorafenib as an active ingredient.

[0036] In one embodiment of the present invention, the composition may additionally include, but is not limited to, sorafenib.

[0037] The present invention provides a kit for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

[0038] The present invention provides a kit for preventing or treating cancer, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

[0039] In one embodiment of the present invention, the kit may further comprise, but is not limited to, sorafenib.

[0040] In one embodiment of the present invention, the cancer may be at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC), but is not limited thereto.

[0041] In addition, the present invention provides a method for enhancing the anticancer effect of sorafenib, a method for co-administering with sorafenib, or a method for preventing or treating cancer, comprising a step of administering to a subject in need thereof a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression in a pharmaceutically effective amount.

[0042] In addition, the present invention provides a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, for use in enhancing the anticancer effect of sorafenib, for use in combination with sorafenib, or for use in preventing or treating cancer.

[0043] In addition, the present invention provides a use for providing a composition comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression, a preparation for enhancing the anticancer effect of sorafenib, a preparation for co-administration with sorafenib, or a preparation for preventing or treating cancer.

[0044] In addition, the present invention provides a method for preventing or treating cancer, comprising administering to a subject in need thereof a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib in a pharmaceutically effective amount.

[0045] The present invention also provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib for use in preventing or treating cancer.

[0046] In addition, the present invention provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a use of the composition comprising sorafenib for preparing a preparation for preventing or treating cancer.

[0047]

[0048] A composition for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector as an active ingredient, comprises a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression, which comprises a sequence commonly expressed by an isoform of the SASH1 gene. Since the vector of the present invention enhances the anticancer effect of sorafenib while also having an excellent anticancer effect itself, a composition comprising a SASH1 overexpression vector and / or sorafenib as an active ingredient can be usefully utilized as a substance for preventing or treating cancer.

[0049]

[0050] Figure 1 is a schematic diagram of the SASH1 overexpression vector used in the present invention.

[0051] Figure 2 shows the IC when sorafenib was treated while overexpressing SASH1. 50 This is a graph showing the reduction effect.

[0052] Figure 3 is a graph showing cell growth inhibition activity under SASH1 overexpression treatment and / or sorafenib conditions.

[0053] Figure 4 is a graph showing cell proliferation activity under SASH1 overexpression treatment and / or sorafenib conditions.

[0054]

[0055] SASH1 (SAM and SH3 domain-containing protein 1) is a tumor suppressor protein that plays a role in key cellular processes including apoptosis and cell proliferation, but these cellular processes are frequently disrupted in human tumors. Despite this, little is known about the role of SASH1 in disease pathogenesis.

[0056] However, the inventors of the present invention completed the present invention by confirming that the cell death (cell growth inhibition) effect and cell proliferation inhibition effect of sorafenib were significantly and excellently increased when a vector overexpressing a gene sequence commonly expressed in the genetic isoform of SASH1 was treated.

[0057] The present invention provides a composition for enhancing the anticancer effect of sorafenib, comprising as an active ingredient a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression.

[0058] In the present invention, the term "polynucleotide sequence for inducing overexpression" may refer to a DNA sequence essential for overexpression of an operably linked nucleotide sequence in a specific host organism. In the present invention, the term "polynucleotide sequence for inducing SASH1 overexpression" may refer to a sequence essential for overexpression of SASH1 in a host cell through a vector or the like containing the sequence.

[0059] In the present invention, the polynucleotide sequence for inducing overexpression may include a promoter for performing overexpression transcription, an optional operator sequence for regulating transcription, a sequence encoding a suitable mRNA ribosome binding site, and a sequence for regulating the termination of transcription and translation. For example, a regulatory sequence suitable for prokaryotes includes a promoter, optionally an operator sequence, and a ribosome binding site. For eukaryotic cells, a promoter, a polyadenylation signal, and an enhancer may be included. The factor that most influences the amount of gene expression in a plasmid is the promoter. As promoters for high expression, the SRα promoter and a cytomegalovirus-derived promoter can be preferably used, but are not limited thereto.

[0060] In the present invention, “overexpression” may mean a higher level of expression than the level at which the corresponding gene is expressed in a cell under a normal state, but is not limited thereto, and may include the broadest meaning meaning a state in which the level of expression of a gene is increased, which is generally applied in the art for pharmaceutical or anticancer promotion purposes.

[0061] In the present invention, “sorafenib” may be an anticancer drug sold under the brand name Nexavar, which is approved for the treatment of primary kidney cancer (advanced renal cell carcinoma), advanced primary liver cancer (hepatocellular carcinoma), FLT3-ITD positive AML, and radioactive iodine-resistant advanced thyroid cancer.

[0062] Sorafenib is a protein kinase inhibitor active against many protein kinases, including VEGFR, PDGFR, and RAF kinases. Among RAF kinases, sorafenib may be more selective for c-Raf than B-RAF. Sorafenib treatment may inhibit tumor growth by inducing autophagy. It is also known to be a potent soluble epoxide hydrolase inhibitor, and this activity is likely to reduce the severity of side effects.

[0063] It is known to be the most effective single-drug therapy, especially for liver cancer, but it has fatal limitations such as high recurrence rate and resistance, and it is known that a treatment strategy to improve drug response is needed.

[0064] As used herein, "vector" means a DNA construct containing a DNA sequence operably linked to suitable regulatory sequences capable of expressing the DNA in a suitable host. The vector may be a plasmid, a phage particle, or simply a potential genomic insert. Once transformed into a suitable host, the vector can replicate and function independently of the host genome, or in some cases, can integrate into the genome itself. Since plasmids are currently the most commonly used form of vector, the terms "plasmid" and "vector" are sometimes used interchangeably herein. However, the present invention encompasses other forms of vectors known or becoming known in the art that have equivalent functionality. Typical expression vectors for mammalian cell culture expression include, but are not limited to, pRK5 (EP 307,247), pSV16B (WO 91 / 08291), and pVL1392 (Pharmingen).

[0065] In one embodiment of the present invention, the polynucleotide sequence for inducing SASH1 overexpression may include, but is not limited to, a sequence commonly expressed in a SASH1 gene isoform.

[0066] In the present invention, the term "isoform" may refer to, but is not limited to, mRNAs that are produced from the same genetic locus but have different transcription start sites (TSSs), protein-coding DNA sequences (CDSs), and / or untranslated regions (UTRs), thereby potentially altering gene function, i.e., different RNAs that can be synthesized from a single gene. Such isoforms can be produced as mRNAs composed of various combinations of introns and exons from the same sequence through a splice variant system.

[0067] In one embodiment of the present invention, the commonly expressed sequence may be commonly expressed in exon 20 of the SASH1 gene homolog, but is not limited thereto.

[0068] In one embodiment of the present invention, the commonly expressed sequence may include, but is not limited to, a polynucleotide sequence of 500 bp to 3,750 bp in length of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0069] In one embodiment of the present invention, the commonly expressed sequence may include, but is not limited to, a portion of exon 20 of the SASH1 gene.

[0070] In one embodiment of the present invention, exon 20 may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 4.

[0071] In one embodiment of the present invention, exon 20 may include, but is not limited to, the 4,199th to 7,949th bases from the 5' end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0072] In one embodiment of the present invention, a portion of the exon 20 sequence may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 1.

[0073] In one embodiment of the present invention, a portion of the sequence of exon 20 may include, but is not limited to, bases 6,608 to 7,108 from the 5' end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

[0074] In one embodiment of the present invention, the vector or the polynucleotide sequence for inducing SASH1 overexpression of the present invention may include the polynucleotide sequence of SEQ ID NO: 1, but is not limited thereto.

[0075] In one embodiment of the present invention, the vector or the polynucleotide sequence for inducing SASH1 overexpression of the present invention may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 2.

[0076] In one embodiment of the present invention, the vector or the polynucleotide sequence for inducing SASH1 overexpression of the present invention may include, but is not limited to, the polynucleotide sequence of SEQ ID NO: 3.

[0077] In the present invention, the cancer is not limited to any cancer for which sorafenib can be used as an anticancer agent. For example, cancers for which sorafenib can be used as an anticancer agent include cancers for which it is used as an adjuvant treatment even if it is not the primary treatment. Furthermore, it may include cancers for which it is used in combination with other treatments, i.e., as an adjuvant treatment, rather than as a single treatment. Such cancers for which sorafenib is applicable include not only those approved by an authorized institution for use as the primary treatment for a specific cancer, but also those types of cancers commonly used in the art. Furthermore, the term "cancer" is not limited to the timing, dose, or duration of sorafenib treatment in the cancer, and may refer to a broad range of cancer types. In one embodiment of the present invention, the cancer may be at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC), but is not limited thereto.

[0078] In one embodiment of the present invention, the composition may be administered in combination with sorafenib.

[0079] In the present invention, “combination administration” can be achieved by administering individual components of the treatment regimen simultaneously, sequentially, or individually. It is a method of obtaining a combination treatment effect by administering two or more drugs simultaneously or sequentially, or alternately at regular or indefinite intervals, etc. Combination therapy is not limited thereto, but can be defined as a combination therapy that provides a synergistic effect while being therapeutically superior to the efficacy obtained by administering one or the other of the components of the combination therapy at a regular dose, as measured by, for example, the degree of response, the rate of response, the period until disease progression, or the period of survival.

[0080] Additionally, single or multiple dosing regimens may be used for combination administration. Considering all of the above factors, those skilled in the art can administer the optimal dose, with the minimum amount necessary to achieve maximum efficacy without adverse effects, under optimal conditions such as recovery, concentration, and administration method.

[0081] In the present invention, “enhancing anticancer effect” refers to all effects that can ultimately strengthen the function of an anticancer agent, and is a concept that includes not only enhancing the anticancer effect of an anticancer agent, such as suppressing tumor growth, suppressing tumor metastasis, and suppressing tumor recurrence, but also increasing or increasing the responsiveness and / or sensitivity of an individual to an anticancer agent by suppressing the formation of resistance or tolerance in cancer cells to the anticancer agent, thereby ultimately enhancing the anticancer effect. In addition, it may include extending the remaining lifespan of a cancer patient, alleviating pain, and improving the well-being of the patient’s life while fighting the disease. In addition, it may include increasing the therapeutic effect that can be generally used, such as increasing the therapeutic synergy effect with other drugs, and reducing the dosage / cycle of the drug. In addition, the present invention may include, but is not limited to, all effects such as generating the same or greater anticancer effect even when using a lower concentration or smaller amount of an anticancer agent compared to the concentration or amount used in the art, generating a therapeutic effect at a concentration and / or dose that did not previously show a therapeutic effect, or generating a therapeutic synergy effect with other drugs that did not show, or even if it did, reducing the concentration and / or dose or increasing the cycle. That is, the composition according to the present invention can be used as a compound for combination administration with known anticancer agents for the purpose of enhancing anticancer effects. That is, the composition of the present invention can be used for combination administration with anticancer agents, thereby enhancing the anticancer effects of said anticancer agents.

[0082]

[0083] The vector of the present invention can be delivered by being included in a delivery vehicle according to a method common in the art, and for example, liposomes can be used, but are not limited thereto.

[0084] The polynucleotide sequence for inducing SASH1 overexpression of the present invention, and the vector sequence including the sequence, may each include a polynucleotide sequence (base sequence) represented by SEQ ID NO: 1 to SEQ ID NO: 3, but are not limited thereto, and variants of the polynucleotide sequence are included within the scope of the present invention. The nucleic acid molecule of the polynucleotide sequence represented by SEQ ID NO: 1 to SEQ ID NO: 5 of the present invention is a concept that includes a functional equivalent of the nucleic acid molecule constituting the same, for example, a variant that can perform the same function functionally as the nucleic acid molecule, although a part of the polynucleotide sequence of the nucleic acid molecule has been modified by deletion, substitution, or insertion. Specifically, the gene may include a polynucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, still more preferably 90% or more, and most preferably 95% or more, with the polynucleotide sequence of the polynucleotide sequence represented by SEQ ID NO: 1 to SEQ ID NO: 5, respectively. For example, it includes polynucleotides having sequence homology of 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%.

[0085] The “% sequence identity” for a polynucleotide is determined by comparing two optimally aligned sequences over a comparison region, where portions of the polynucleotide sequence in the comparison region may contain additions or deletions (i.e., gaps) compared to the reference sequence (which does not contain additions or deletions) for the optimal alignment of the two sequences.

[0086] In the present invention, sequence number 4 represents a polynucleotide sequence of exon 20 of the SASH1 gene, and the bold text of sequence number 4 in Table 1 represents a polynucleotide sequence for inducing SASH1 overexpression, which may be sequence number 1.

[0087] In addition, in the present invention, sequence number 5 is the SASH1 gene, and the polynucleotide sequence of exon 20 is underlined in Table 1 and is the same as sequence number 4. In addition, bold text may be sequence number 1.

[0088]

[0089] The present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0090] The present invention provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a pharmaceutical composition for preventing or treating cancer comprising sorafenib as an active ingredient.

[0091] In one embodiment of the present invention, the composition may additionally include, but is not limited to, sorafenib.

[0092] In the present invention, the composition comprising a vector and sorafenib does not necessarily mean only a composition comprising the vector and sorafenib simultaneously, but may have the broadest meaning including a composition comprising each composition included in a solvent, formulation, etc. to best exhibit each effect.

[0093] The oligopeptide according to the present invention or its analogue can be used for the prevention and / or treatment of cancer. The term "cancer" as used herein refers to a disease characterized by uncontrolled cell growth, which results in the formation of a tumor, a mass of cells, which invades surrounding tissues and, in severe cases, metastasizes to other organs of the body. Academically, it is also called a neoplasm. Cancer is an intractable chronic disease that often fails to achieve a fundamental cure even with surgery, radiation, and chemotherapy, causing suffering to patients and ultimately leading to death. There are various factors that contribute to the development of cancer, but these are categorized as intrinsic and extrinsic. While the precise mechanism by which normal cells transform into cancer cells is not yet fully understood, it is known that a significant number of cancers are caused by external factors, such as environmental factors. Internal factors include genetic factors and immunological factors, while extrinsic factors include chemicals, radiation, and viruses. Genes involved in the development of cancer include oncogenes and tumor suppressor genes, and cancer occurs when the balance between these genes is disrupted by the internal or external factors described above.

[0094] The cancer may be a solid cancer or a blood cancer, and may be at least one selected from the group consisting of, but not limited to, squamous cell carcinoma, lung cancer, adenocarcinoma of the lung, peritoneal cancer, skin cancer, cutaneous or intraocular melanoma, rectal cancer, anal cancer, esophageal cancer, small intestinal cancer, endocrine cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, blood cancer, liver cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, hepatoma, breast cancer, colon cancer, large intestine cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, brain cancer, and more specifically, may be a cancer selected from the group consisting of lung cancer, breast cancer, blood cancer, colon cancer, pancreatic cancer, and combinations thereof.

[0095] In the present invention, the lung cancer may be non-small cell lung carcinoma or lung papillary adenocarcinoma. In one embodiment of the present invention, the compound according to the present invention is used in patients with T790M mutation positive, EGFR m - It may be for the treatment of lung cancer in patients, and / or patients who have developed resistance to osimertinib, but is not limited thereto. In addition, in the present invention, the lung cancer may include, but is not limited to, EGFR mutations (e.g., L858R mutation, and / or T790M mutation) and / or MET amplification mutations. The present inventors have confirmed through specific examples that when the compound according to the present invention is used in combination with an anticancer agent, an excellent anticancer effect is exhibited against cancer cells or animal models containing the mutations.

[0096] In addition, the breast cancer may be, but is not limited to, hormone receptor (HR)-positive breast cancer. In addition, the breast cancer may be triple-negative breast cancer. In addition, the breast cancer according to the present invention may include, but is not limited to, a BRCA1 mutation (e.g., a 5382C mutation). The inventors of the present invention confirmed through specific examples that when the compound according to the present invention is used in combination with an anticancer agent, an excellent anticancer effect is exhibited against cancer cells or animal models containing the mutation.

[0097] Additionally, the blood cancer may be leukemia, lymphoma, multiple myeloma, etc.

[0098] In the present invention, the effect of preventing and / or treating cancer includes not only the effect of inhibiting the growth of cancer cells, but also the effect of inhibiting the worsening of cancer due to migration, invasion, metastasis, etc.

[0099] The composition for preventing or treating cancer according to the present invention has the same meaning as a pharmaceutical composition for preventing or treating cancer, and may further include suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions. The excipients may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film-coating materials, and controlled-release additives.

[0100] The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, ellipsoids, emulsions, suspensions, alcohols, troches, aromatic waters, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, ointments, lotions, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols, according to a conventional method, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasmas.

[0101] Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0102] When formulating, it is usually prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

[0103] The additives of the tablets, powders, granules, capsules, pills, and troches according to the present invention include excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl alcohol, polyvinyl pyrrolidone, and binders such as hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, Disintegrants such as carboxymethylcellulose calcium, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, baking soda, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethylcellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, and light anhydrous silicic acid;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium dentata, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid can be used.

[0104] As additives of the liquid formulation according to the present invention, water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, etc. can be used.

[0105] The syrup according to the present invention may include a solution of white sugar, other sugars, or sweeteners, and may also include a fragrance, a coloring agent, a preservative, a stabilizer, a suspending agent, an emulsifier, a viscosity modifier, and the like, as needed.

[0106] Purified water may be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. may be used as needed.

[0107] The suspension according to the present invention may include suspending agents such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, and the like. Surfactants, preservatives, stabilizers, colorants, and fragrances may also be used as needed.

[0108] The injection according to the present invention includes a solvent such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, benzene benzoate; a solubilizing agent such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tween, nitrile acid amide, hexamine, dimethylacetamide; a buffer such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumins, peptones, gums; It may include isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; oxidizing agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid, and acetone sodium bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium cis-methylenediamine, sodium alginate, Tween 80, and aluminum monostearate.

[0109] The suppository according to the present invention comprises cocoa butter, lanolin, withepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, ranet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydroxycote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Hydroxycote Mechanisms such as (Hydrokote) 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository base type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecovi (W, R, S, M, Fs), Tezester triglyceride base (TG-95, MA, 57) can be used.

[0110] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

[0111] Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0112] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the patient’s disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the excretion rate, the treatment period, the concurrently used drugs, and other factors well known in the medical field. A preferred dosage can be selected according to the condition and body weight of the subject, the degree of the disease, the form of the drug, the route of administration, and the period. As a specific example, the pharmaceutical composition can be administered once or several times a day in an amount of 0.001 to 1000 mg / kg, 0.01 to 100 mg / kg, 0.01 to 10 mg / kg, 0.1 to 10 mg / kg, or 0.1 to 1 mg / kg.

[0113] The pharmaceutical composition according to the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents. It can be administered sequentially or simultaneously with conventional therapeutic agents, or in single or multiple doses. Taking all of the above factors into account, it is important to administer an amount that achieves maximum efficacy with minimal side effects. This amount can be readily determined by those skilled in the art to which the present invention pertains.

[0114] The pharmaceutical composition of the present invention can be administered to a subject via various routes. All modes of administration are conceivable, including oral ingestion, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, intrathecal injection, sublingual administration, buccal mucosa administration, rectal insertion, vaginal insertion, ocular administration, otic administration, nasal administration, inhalation, oral or nasal spraying, dermal administration, and transdermal administration.

[0115] The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with various related factors such as the disease to be treated, route of administration, age, sex, weight of the patient, and severity of the disease.

[0116] In the present invention, “subject” means a subject requiring treatment for a disease, and more specifically, means a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, and cow.

[0117] In the present invention, “administration” means providing a predetermined composition of the present invention to an individual by any appropriate method.

[0118] In the present invention, “prevention” means any action that suppresses or delays the onset of a target disease, “treatment” means any action that improves or beneficially changes a target disease and its metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and “improvement” means any action that reduces a parameter related to a target disease, for example, the severity of a symptom, by administering a composition according to the present invention.

[0119]

[0120] The present invention provides a composition for co-administration with sorafenib, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0121] In addition, the composition of the present invention may be formulated with sorafenib and administered simultaneously, separately, or sequentially. In this case, the composition may be a pharmaceutical composition for combination administration for simultaneous or sequential administration, comprising a first pharmaceutical composition containing a pharmaceutically effective amount of the vector of the present invention as an active ingredient; and a second pharmaceutical composition containing a pharmaceutically effective amount of the anticancer agent; sorafenib as an active ingredient. In this case, in the case of sequential administration, the administration order is not limited, and the administration regimen may be appropriately adjusted depending on the patient's condition, etc.

[0122] That is, when the pharmaceutical composition is a combination-administration pharmaceutical composition for sequential administration, the composition may be administered first with the composition containing the vector of the present invention as an active ingredient (“first component”), and then with the anticancer agent; sorafenib (“second component”), or the reverse order is also possible.

[0123] Alternatively, the present invention provides an anticancer adjuvant pharmaceutical composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

[0124] The anticancer adjuvant pharmaceutical composition of the present invention may mean, but is not limited to, a composition that causes or enhances an anticancer effect when administered in combination with an anticancer agent.

[0125] In addition, in the present invention, “anticancer effect” may mean the effect of the anticancer agent itself, or causing or enhancing responsiveness to the anticancer agent, or may mean causing or enhancing the effect of combination treatment with other anticancer agents or anticancer treatment methods.

[0126] In addition, in the present invention, “anticancer adjuvant” means something that helps to increase the above-described “anticancer effect” of the anticancer agent, and may include everything that is the same as or assists the effect of “anticancer effect enhancement” of the present invention.

[0127] The anticancer adjuvant pharmaceutical composition of the present invention can be administered in combination with an anticancer agent, and the timing, cycle, dosage, concentration, method, etc. of the combination can be administered in the most preferable manner taking into account the clinical condition of the patient to whom the composition is administered, the type of cancer, the type of anticancer agent to be administered in combination, the type of other compositions, etc.

[0128] Additionally, the anticancer adjuvant pharmaceutical composition may be prepared together with other substances that may generally be added to enhance the adjuvant effect of the anticancer agent, and may be administered in combination with other compositions.

[0129] In addition, pretreatment or post-treatment methods, storage methods, and usage methods generally performed to enhance auxiliary effects can be applied without limitation.

[0130]

[0131] The present invention provides a kit for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

[0132] The present invention provides a kit for preventing or treating cancer, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

[0133] In one embodiment of the present invention, the kit may further comprise, but is not limited to, sorafenib.

[0134] In one embodiment of the present invention, the cancer may be at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC), but is not limited thereto.

[0135] In the present invention, the “kit” means a tool that enhances the anticancer effect of sorafenib by using a vector including a polynucleotide sequence for inducing SASH1 overexpression of the present invention, or that can prevent or treat cancer by including the vector of the present invention and sorafenib. In addition to the above substances, the kit of the present invention may include other components, compositions, solutions, devices, etc. that are usually necessary for methods of storing and processing them. As a specific example, in this case, each component may be applied once or more without limitation in the number of times, there is no limitation on the order in which each substance is applied, and the application of each substance may be performed simultaneously or microscopically.

[0136] In the present invention, the kit may include a container; instructions; and the like. The container may serve to package the substance, and may also serve to store and fix the substance. The material of the container may take the form of, for example, a bottle, a tub, a sachet, an envelope, a tube, an ampoule, and the like, and these may be formed partially or wholly from plastic, glass, paper, foil, wax, and the like. The container may be initially equipped with a completely or partially detachable stopper, which may be part of the container or may be attached to the container by mechanical, adhesive, or other means, and may also be equipped with a stopper for allowing access to the contents by means of a syringe needle. The kit may include an outer package, and the outer package may include instructions for the use of the components.

[0137]

[0138] In addition, the present invention provides a method for enhancing the anticancer effect of sorafenib, a method for co-administering with sorafenib, or a method for preventing or treating cancer, comprising a step of administering to a subject in need thereof a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression in a pharmaceutically effective amount.

[0139] In addition, the present invention provides a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, for use in enhancing the anticancer effect of sorafenib, for use in combination with sorafenib, or for use in preventing or treating cancer.

[0140] In addition, the present invention provides a use for providing a composition comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression, a preparation for enhancing the anticancer effect of sorafenib, a preparation for co-administration with sorafenib, or a preparation for preventing or treating cancer.

[0141] In addition, the present invention provides a method for preventing or treating cancer, comprising administering to a subject in need thereof a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib in a pharmaceutically effective amount.

[0142] The present invention also provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib for use in preventing or treating cancer.

[0143] In addition, the present invention provides a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a use of the composition comprising sorafenib for preparing a preparation for preventing or treating cancer.

[0144]

[0145] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples.

[0146]

[0147] [Example]

[0148]

[0149] [Materials and Methods]

[0150]

[0151] Cell line preparation

[0152] HepG2.2.15 cells were donated by the Korea Advanced Institute of Science and Technology (KAIST, Daejeon, Korea). Cells were cultured in Dulbecco's Modified Eagle's medium (DMEM; Hyclone, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen Life Technologies, USA) at 37°C in a humidified incubator containing 5% CO2.

[0153]

[0154] Cell transfection: inducing overexpression

[0155] Cell transfection was performed using Lipofectamine in Opti-MEM according to the manufacturer's instructions. ® 3000 (Invitrogen, L3000015) was used. Specifically, diluted Lipofectamine ® After mixing 3000 with the vector, the mixture was incubated at room temperature for 10 minutes. The mixture was then added to a HepG2.2.15 cell culture plate and incubated at 37°C in an atmosphere containing 5% CO2.

[0156]

[0157] Example 1. Construction of SASH1 overexpression vector

[0158] The SASH1 overexpression vector of the present invention was constructed. Specifically, in order to construct the SASH1 overexpression vector of the present invention, the exon sequence shared by the transcriptional isoforms of SASH1 was investigated. At this time, the sequence was confirmed using the mRNA of human SASH1 domain, transcript variant X1 (Homo sapiens SAM and SH3 domain containing 1 (SASH1), transcript variant X1, mRNA, NCBI XM_054354867.1).

[0159] Among the 20 exons included in SASH1, the exons commonly expressed by all gene isoforms were confirmed to be exon 10 (347 bp), exon 11 (75 bp), exon 12 (144 bp), exon 13 (136 bp), exon 14 (170 bp), exon 15 (210 bp), exon 16 (151 bp), exon 17 (114 bp), exon 18 (1,139 bp), exon 19 (132 bp), and exon 20 (3,750 bp). Among them, a vector was constructed using a sequence (SEQ ID NO: 2, 516 bp) including a portion (SEQ ID NO: 1, 6,608 bp to 7,108 bp of SASH1 gene SEQ ID NO: 3, total 500 bp) of exon 20 (from 4,199 bp to 7,949 bp of SASH1 gene SEQ ID NO: 3, total 3,750 bp) (Fig. 1). The vector of the present invention is represented by SEQ ID NO: 3 in Table 1, and at this time, the 5' end was constructed by cutting with HindIII and the 3' end with XbaI.

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170] Example 2. Confirmation of the cell death activity enhancement effect of sorafenib due to SASH1 overexpression.

[0171] The cell death effect was analyzed when the SASH1 overexpression vector manufactured according to Example 1 and sorafenib were treated separately or together. Specifically, HepG2.2.15 cells were cultured at 1×10 per well. 4 Each cell was cultured in a 96-well plate, and the drug was treated so that a total of 100 μl per well was made up, and the remaining amount was filled with medium. Then, 2.3 μl or 4.5 μl per well was treated with sorafenib diluted in DMSO to a concentration of 0.01 μM, 0.1 μM, 0.5 μM, 1 μM, 2 μM, 4 μM, 8 μM, 10 μM, and 20 μM. In addition, 1 μg of SASH1 overexpression vector was transfected in liposome form using lipofectamine 3000 (Invitrogen, L3000015) at 100 μl per well. As a negative control, the same volume of DMSO or empty vector (pcDNA) was treated and used. After 24 hours, 10 μl of CCK-8 reagent was added per well, and after 2 hours, the medium was removed and the absorbance was measured.

[0172]

[0173] As a result (Fig. 2), in the experimental group treated with sorafenib alone, the IC of sorafenib 50 was measured as 4.3 μM. In addition, in the experimental group treated with sorafenib while overexpressing SASH1, the IC of sorafenib 50 It was found that the concentration decreased to 2.9 μM, confirming a reduction effect of approximately 30%.

[0174]

[0175] These results confirmed that overexpression of SASH1 enhances the apoptotic activity of sorafenib, as the same apoptotic effect could be achieved with only a small amount of sorafenib treatment by overexpressing SASH1.

[0176]

[0177] Example 3. Confirmation of the cell growth inhibition effect according to SASH1 overexpression and the cell growth inhibition activity enhancement effect of sorafenib.

[0178] The cell growth inhibition effect of the SASH1 overexpression vector produced according to Example 1 and sorafenib when treated separately or together was analyzed. Specifically, HepG2.2.15 cells were cultured at 1×10 per well. 4 Each cell was cultured in a 96-well plate, and the drug was treated so that a total of 100 μl per well was made up, and the remaining amount was filled with medium. Sorafenib diluted in DMSO was treated at 5.6 μl per well to a concentration of 3.5 μM. In addition, 1 μg of SASH1 overexpression vector was transfected in liposome form at 100 μl per well using lipofectamine 3000 (Invitrogen, L3000015). The same volume of DMSO or empty vector (pcDNA) was used as a negative control. After 24 hours, 10 μl of CCK-8 reagent was added per well, and after 2 hours, the medium was removed and the absorbance was measured.

[0179]

[0180] As a result (Fig. 3), the experimental group overexpressing SASH1 showed a 35% decrease in cell growth compared to the negative control group, indicating that overexpression of SASH1 alone can inhibit cell growth, and this was confirmed to be statistically significant.

[0181] In addition, the experimental group treated with sorafenib alone showed a 40% decrease in cell growth compared to the negative control group, and the experimental group treated with sorafenib while overexpressing SASH1 showed an 80% decrease in cell growth compared to the negative control group. Therefore, it was confirmed that the cell growth inhibition activity of treating with sorafenib while overexpressing SASH1 was twice as high as the cell growth inhibition activity of treating with sorafenib alone or overexpressing SASH1, showing a significantly superior synergistic effect.

[0182]

[0183] Example 4. Confirmation of the cell proliferation inhibition effect according to SASH1 overexpression and the cell proliferation inhibition activity enhancement effect of sorafenib.

[0184] The cell proliferation inhibition effect was analyzed when the SASH1 overexpression vector manufactured according to Example 1 and sorafenib were treated separately or together. Specifically, HepG2.2.15 cells were cultured at 1×10 per well. 4 Each cell was cultured in a 96-well plate, and the drug was treated so that a total of 100 μl per well was filled with medium. 5.6 μl of sorafenib diluted in DMSO was treated per well to achieve a concentration of 3.5 μM. In addition, 1 μg of SASH1 overexpression vector was transfected in liposome form at 100 μl per well using lipofectamine 3000 (Invitrogen, L3000015). An equal volume of DMSO or empty vector (pcDNA) was used as a negative control. After 24 hours, the cells were washed with distilled water and fixed in 4% formalin for 15 minutes. Afterwards, the cells were stained with 0.1% crystal violet, washed with tap water, and the wells were photographed. The area was calculated using imageJ software, and the growth rate of each experimental group was compared with the negative control group.

[0185]

[0186] As a result (Fig. 4), the experimental group overexpressing SASH1 and the experimental group treated with sorafenib alone showed a 40% decrease in cell proliferation compared to the negative control group. At this time, the cell proliferation effect when SASH1 was overexpressed alone was the same as the experimental group treated with sorafenib alone, and this corresponds to a statistically significant effect, so these results suggest that cell proliferation activity is excellent even when SASH1 is overexpressed alone.

[0187] In addition, the experimental group treated with sorafenib while overexpressing SASH1 showed a 75% decrease in cell growth compared to the negative control group, indicating that it exhibited approximately 1.9 times more cell proliferation inhibition activity than when sorafenib was treated alone or when SASH1 was overexpressed alone.

[0188]

[0189] In conclusion, when SASH1 is overexpressed, not only is the cell growth inhibition activity and cell proliferation inhibition activity excellent even when treated alone, but it also exhibits a remarkably excellent synergy effect on the anticancer activity of sorafenib. Therefore, it is expected that the SASH1 overexpression agent will be usefully utilized as an anticancer composition or a composition for enhancing the anticancer effect of sorafenib.

[0190]

[0191] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0192]

[0193] A composition for enhancing the anticancer effect of sorafenib, comprising the SASH1 overexpression vector of the present invention as an active ingredient, comprises a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression, which comprises a sequence commonly expressed by an isoform of the SASH1 gene. Since the vector of the present invention increases the anticancer effect of sorafenib while also having an excellent anticancer effect itself, a composition comprising the SASH1 overexpression vector and / or sorafenib as an active ingredient can be usefully utilized as a substance for preventing or treating cancer, and thus its industrial applicability is recognized.

Claims

1. A composition for enhancing the anticancer effect of sorafenib, comprising as an active ingredient a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 (SAM and SH3 domain-containing protein 1) overexpression.

2. In paragraph 1, A composition wherein the polynucleotide sequence for inducing the above SASH1 overexpression comprises a sequence commonly expressed in a SASH1 gene isoform.

3. In paragraph 2, A composition wherein the above commonly expressed sequence is commonly expressed in exon 20 of the SASH1 gene isoform.

4. In paragraph 3, A composition wherein the commonly expressed sequence comprises a portion of exon 20 of the SASH1 gene.

5. In paragraph 4, A composition, wherein the commonly expressed sequence comprises a polynucleotide sequence of 500 bp to 3,750 bp in length of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

6. In paragraph 4, A composition wherein the above exon 20 comprises a polynucleotide sequence of SEQ ID NO:

4.

7. In paragraph 5, A composition, wherein the above exon 20 comprises bases 4,199 to 7,949 from the 5′ end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

8. In paragraph 4, A composition, wherein a portion of the above exon 20 sequence comprises a polynucleotide sequence of SEQ ID NO:

1.

9. In paragraph 4, A composition, wherein a portion of the above exon 20 sequence includes bases 6,608 to 7,108 from the 5′ end of the SASH1 gene (NCBI Reference Sequence: XM_054354867.1).

10. In paragraph 1, A composition wherein the above vector comprises a polynucleotide sequence of sequence number 1.

11. In paragraph 1, A composition wherein the above vector comprises a polynucleotide sequence of sequence number 2.

12. In paragraph 1, A composition wherein the above vector comprises a polynucleotide sequence of sequence number 3.

13. In paragraph 1, A composition wherein the cancer is at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC).

14. A composition for co-administration with sorafenib, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

15. A pharmaceutical composition for adjuvant anticancer treatment, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

16. A pharmaceutical composition for preventing or treating cancer, comprising a SASH1 overexpression vector including a polynucleotide sequence for inducing SASH1 overexpression as an active ingredient.

17. In paragraph 16, A pharmaceutical composition, wherein the composition further comprises sorafenib.

18. A SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a pharmaceutical composition for preventing or treating cancer, comprising sorafenib as an active ingredient.

19. A kit for enhancing the anticancer effect of sorafenib, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and an instruction manual.

20. A kit for preventing or treating cancer, comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and instructions.

21. In paragraph 19 or 20, A kit further comprising sorafenib.

22. In paragraph 19 or 20, A kit wherein the cancer is at least one selected from the group consisting of hepatocellular carcinoma (HCC), advanced thyroid cancer, and renal cell carcinoma (RCC).

23. A method for enhancing the anticancer effect of sorafenib, comprising the step of administering to a subject in need thereof a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression in a pharmaceutically effective amount.

24. A method for preventing or treating cancer, comprising administering to a subject in need thereof a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression in a pharmaceutically effective amount.

25. A method of co-administration with sorafenib, comprising the step of administering to a subject in need thereof a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression in a pharmaceutically effective amount.

26. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression for enhancing the anticancer effect of sorafenib.

27. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression for use in preventing or treating cancer.

28. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, for use in combination with sorafenib.

29. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression to provide a preparation for enhancing the anticancer effect of sorafenib.

30. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, for use in combination with sorafenib.

31. Use of a composition comprising a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression to provide a preparation for preventing or treating cancer.

32. A method for preventing or treating cancer, comprising administering to a subject in need thereof a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib in a pharmaceutically effective amount.

33. Use of a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib for the prevention or treatment of cancer.

34. Use of a SASH1 overexpression vector comprising a polynucleotide sequence for inducing SASH1 overexpression, and a composition comprising sorafenib for the preparation of a preparation for preventing or treating cancer.

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