Pharmaceutical composition for preventing or treating tumor disease containing auranofin as active ingredient

Abstract

To provide a new pharmaceutical composition for preventing or treating neoplasticdisorders, and to provide a method for preventing or treating neoplasticdisorders using the composition.SOLUTION: [Technical Field] The present invention relates to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and more particularly, to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and a method for preventing or treating tumor diseases using the composition containing auranofin.SELECTED DRAWING: Figure 5B

Description

[Technical Field]

[0001] This invention was made under the support of the Ministry of Science and ICT under the project specific number 1711197632 and detailed project number 00239096, the research management organization for this project is the Korea Research Foundation, the research project name is "Individual Basic Research (Ministry of Science and ICT)", the research topic name is "Discovery and innovative research into therapeutic agents for acoustic neuroma using drug regeneration", the sponsoring organization is Soonchunhyang University, and the research period is from 2023.06.01 to 2024.02.29.

[0002] This patent application claims priority to Korean Patent Application No. 10-2024-0091599, filed with the Korean Intellectual Property Office on July 11, 2024, the disclosure of which is incorporated herein by reference.

[0003] The present invention relates to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and more particularly to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and a method for preventing or treating tumor diseases using a composition containing auranofin. [Background technology]

[0004] The human body has 12 pairs of nerves originating from the brain and 31 pairs of nerves originating from the spinal cord. These nerves connect to various parts of the body through passages in the skull and vertebrae. Schwannoma is a collective term for tumors that originate in the nerve sheath, or Schwann cells, which are the crown-like structures that surround and support these nerves. Schwannomas usually develop in the narrow spaces where nerves exit the brain or spinal cord and enter the bone passages, but they can also develop in peripheral nerves and soft tissues. When they originate in cranial nerves, they manifest as brain tumors. Since 70-80% of brain tumors originate in the auditory nerve, they are commonly called acoustic neurinomas. When they originate in the spinal cord, they are called spinal neurinomas. While brain tumors can rarely be malignant, most are benign, and if complete removal is possible, they can be treated with surgery alone. Schwannoma tumor cells grow slowly and rarely metastasize to organs other than the brain. However, they can recur even after complete removal. Furthermore, if the tumor grows larger or invades surrounding brain tissue, cranial nerves, or cerebral blood vessels, it becomes difficult to remove even through surgery, and there is a high chance of complications occurring during surgery. It accounts for around 8% of all brain tumors, including malignant ones, and is more common in women than men, but occurs most frequently in people in their 30s to 60s.

[0005] Neurofibromatosis (NF) is a genetic disorder affecting bone, soft tissue, skin, and the nervous system. It is classified as type 1 (NF1) or type 2 (NF2). NF2 is a benign tumor that develops in the cranial nerve (the eighth nerve) and presents with symptoms of hearing loss, tinnitus, and balance problems. It is also called vestibular schwannomas because they arise within Schwann cells in the central nervous system. The average age at onset of NF2 is 18-24 years, and by the age of 30, most patients develop bilateral vestibular schwannomas. Furthermore, schwannomas of cranial and peripheral nerves, meningiomas, ventricular theca cells, and, rarely, astrocytomas, can develop.

[0006] Drug repositioning is a method of confirming the potential of new disease treatments for drugs that are already on the market and whose safety has been proven, or drugs whose safety has been verified but which have not passed clinical trials due to efficacy issues. This not only shortens the development period for new drugs, which can take more than 10 years, but also significantly reduces the risks associated with drug safety issues.

[0007] Drug re-creation for acoustic schwannoma, a type of schwannoma, has shown that mifepristrone is effective against the acoustic schwannoma cell line HEI-193, as a result of applying gene expression data confirmed from acoustic schwannoma to FDA-approved drug information using a computer algorithm. Research has also shown that the growth of acoustic schwannoma is suppressed in patients taking aspirin, and clinical trials have been ongoing since 2018 to confirm the effectiveness of aspirin on acoustic schwannoma, targeting 300 patients. However, both drugs require high dosages to be effective against acoustic schwannoma cell lines, and when side effects and toxicity are taken into account, the therapeutic effect is less stable.

[0008] Auranofin (C 20 H 34 Auranofin (AuO9PS) is a gold phosphine compound used to treat rheumatoid arthritis. Recent research has shown that it is effective against brain cancer, a representative malignant tumor that occurs in the brain. However, the effectiveness of auranofin against schwannoma and neurofibromatosis has not yet been confirmed.

[0009] Currently, the main treatments for schwannoma include surgery, radiation therapy, and follow-up observation, with complete surgical removal being the most effective. However, there are limitations depending on the patient's age, general condition, severity of symptoms, tumor size and location, and the risks and complications associated with surgery must also be considered. Although schwannoma is a disease that can cause hearing loss, paresthesia, ataxia, balance disorders, and death due to brainstem compression, it is classified as a benign tumor rather than a cancer, and the prescription of anticancer drugs has not been approved. For this reason, there is an urgent need to develop new therapeutic agents that can provide high therapeutic effects even in small amounts that do not affect the human body. Summary of the Invention [Problem to be solved by the invention]

[0010] Therefore, the present inventors have discovered drugs that are effective against tumor diseases through drug re-creation, and have completed an invention relating to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient.

[0011] Therefore, an object of the present invention is to provide a pharmaceutical composition for preventing or treating neoplastic disorders, which comprises auranofin as an active ingredient.

[0012] Another object of the present invention is to provide a method for preventing or treating a tumor disease, comprising administering to a non-human subject a composition comprising auranofin. [Means for solving the problem]

[0013] The present invention relates to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and more particularly to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient, and a method for preventing or treating tumor diseases using a composition containing auranofin.

[0014] The present invention will now be described in more detail.

[0015] One aspect of the present invention relates to a pharmaceutical composition for preventing or treating neoplastic disorders, which comprises auranofin as an active ingredient.

[0016] The tumor disease may be one or more selected from the group consisting of schwannoma and neurofibromatosis type 2 syndrome, but is not limited thereto.

[0017] The schwannoma may be, but is not limited to, a vestibular schwannoma.

[0018] The composition may contain auranofin at a concentration of 0.1 to 5 mg / mL, preferably 0.1 to 3 mg / mL, 0.1 to 1 mg / mL, 0.2 to 5 mg / mL, and 0.2 to 3 mg / mL, for example, 0.2 to 1 mg / mL, but is not limited to this.

[0019] The auranofin may be administered at a dose of 1 to 20 mg per kg of body weight of the subject, preferably at a dose of 1 to 15 mg, 1 to 10 mg, 2 to 20, or 2 to 15 mg per kg of body weight, for example, at a dose of 2 to 10 mg per kg of body weight, but is not limited thereto.

[0020] The composition may be administered to a subject two to five times per week, preferably two to three times per week, and three to five times per week, for example, three times per week.

[0021] If the composition is administered less than twice a week, the preventive or therapeutic effect will be minimal, and if it is administered more than five times a week, toxicity may occur in the administered subject.

[0022] The pharmaceutical composition of the present invention may be used as a pharmaceutical composition containing a pharmaceutically effective amount of the composite powder and / or a pharmaceutically acceptable carrier.

[0023] As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the efficacy or activity of the composite powder described above.

[0024] The pharmaceutically acceptable carriers contained in the pharmaceutical composition of the present invention are those commonly used in formulations, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc. In addition to these ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc.

[0025] The pharmaceutical composition of the present invention may be administered to mammals, including humans, via various routes, including any commonly used route, such as oral, cutaneous, intravenous, intramuscular, or subcutaneous, preferably oral.

[0026] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, weight, sex, pathological condition, diet of the patient, administration time, administration route, excretion rate and reaction sensitivity, and an ordinary skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention.

[0027] The pharmaceutical compositions of the present invention may be prepared in unit dose form or in multi-dose containers by formulating them with pharmaceutically acceptable carriers and / or excipients by a method readily practiced by those skilled in the art to which this invention pertains. In this case, the dosage form may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablets, capsules, or gel (e.g., hydrogel), and may further contain a dispersing agent or stabilizer.

[0028] Another aspect of the present invention relates to a method for preventing or treating a tumor disease, comprising administering to a subject a composition comprising auranofin.

[0029] The subject may be a mammal, including but not limited to a non-human mammal.

[0030] The tumor disease may be one or more selected from the group consisting of schwannoma and neurofibromatosis type 2 syndrome, but is not limited thereto.

[0031] The schwannoma may be, but is not limited to, a vestibular schwannoma.

[0032] The composition may contain auranofin at a concentration of 0.1 to 5 mg / mL, preferably 0.1 to 3 mg / mL, 0.1 to 1 mg / mL, 0.2 to 5 mg / mL, and 0.2 to 3 mg / mL, for example, 0.2 to 1 mg / mL, but is not limited to this.

[0033] The auranofin may be administered at a dose of 1 to 20 mg per kg of body weight of the subject, preferably at a dose of 1 to 15 mg, 1 to 10 mg, 2 to 20, or 2 to 15 mg per kg of body weight, for example, at a dose of 2 to 10 mg per kg of body weight, but is not limited thereto.

[0034] The composition may be administered to a subject two to five times per week, preferably two to three times per week, and three to five times per week, for example, three times per week. [Effects of the Invention]

[0035] The present invention relates to a pharmaceutical composition for preventing or treating tumor diseases, which contains auranofin as an active ingredient. The composition has excellent tumor growth inhibitory effects against tumor diseases such as schwannoma and neurofibromatosis type 2, and can therefore be used as a pharmaceutical composition for preventing or treating tumor diseases. [Brief explanation of the drawings]

[0036] [Figure 1] 1 is a graph showing the measurement of cytotoxicity against schwannoma cell lines and neurofibromatosis type 2 cell lines by high-throughput screening (HTS) of an FDA-approved drug library according to an embodiment of the present invention. [Figure 2] 1 is a graph showing the IC50 values ​​of drugs discovered by drug re-creation according to an embodiment of the present invention measured against a schwannoma cell line and a neurofibromatosis type 2 cell line. [Figure 3] 1 is a graph showing the measurement and comparison of the cytotoxicity and IC50 values ​​of auranofin, aspirin, and mifepristone against a schwannoma cell line and a neurofibromatosis type 2 cell line according to an embodiment of the present invention. [Figure 4] 1 shows photographs showing changes in expression of cell death markers after treating a schwannoma cell line and a neurofibromatosis type 2 cell line with auranofin according to one embodiment of the present invention. [Figure 5A] 1 shows photographs of mice in which tumor formation was induced according to an embodiment of the present invention, divided into groups according to the dose of auranofin administered. [Figure 5B] 1 is a photograph showing a comparison of tumor size after administration of auranofin to mice in which tumor formation was induced according to an embodiment of the present invention. [Figure 5C] 1 is a graph comparing tumor sizes after administration of auranofin to mice in which tumor formation was induced according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0037] The present invention will be described in more detail below with reference to the following examples, but these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0038] Example 1: Cultivation of Schwannoma Cell Lines and Neurofibromatosis Type 2 Cell Lines Schwannoma cell line RT4-D6P2T (RT4) and neurofibromatosis type 2 cell line SC4 were each 60 mm 2 The cells were cultured on a culture dish, and when they reached about 50% confluency, they were serum starved in FBS-free DMEM medium for 6 hours.

[0039] The RT4 cell line was purchased from the American Type Culture Collection (CRL-2768), and the SC4 cell line was provided by House Ear Institute (Los Angeles, CA, USA). Both RT4 and SC4 cell lines were cultured in DMEM medium containing 10% FBS and 100 units / ml penicillin / streptomycin in a 37°C incubator with 5% CO2.

[0040] Example 2: New drug discovery using drug regeneration To discover effective therapeutic agents for schwannoma and neurofibromatosis type 2 using drug regeneration, high-throughput screening (HTS) was performed on the schwannoma cell line RT4 and neurofibromatosis type 2 cell line SC4 using a library of 2,920 FDA-approved drugs. As shown in Figure 1, RT4 and SC4 cell lines cultured as described in Example 1 were treated with 2,920 drugs at 10 μM each for 24 hours, and then cytotoxicity was measured. Drugs that showed 70% or more cytotoxicity in both cell lines were selected, and eight drugs, including auranofin, were discovered.

[0041] Example 3: Cell viability assay for schwannoma cell lines IC of the eight drugs discovered in Example 2 against the schwannoma cell line RT4 50 The RT4 cell line cultured according to Example 1 was treated with each drug at different concentrations, and the concentration at which the cell survival rate reached 50% after 24 hours was measured to determine the IC 50 The values ​​are shown in Table 1. [Table 1] As can be seen in Figure 2, all eight drugs we discovered had low IC concentrations for the RT4 cell line. 50 In particular, auranofin showed IC 50 The value was 0.23, the lowest compared to other drugs. These results confirmed that all eight drugs were able to inhibit the survival of schwannoma cell lines at low doses.

[0042] Example 4: Comparison of cell viability with existing drugs Among the eight drugs identified, this drug had the lowest IC against the schwannoma cell line RT4. 50 The efficacy of auranofin was further evaluated. RT4 and SC4 cell lines cultured according to Example 1 were treated with auranofin, aspirin, and mifepristone at different concentrations for 24 hours, and the cell viability was compared. The results are shown in Table 2 and Figure 3. [Table 2] Auranofin IC 50 The IC values ​​were 0.25 μM and 0.49 μM in the RT4 and SC4 cell lines, respectively, and the IC values ​​for aspirin and mifepristone were 0.25 μM and 0.49 μM in the RT4 and SC4 cell lines, respectively. 50 was found to be significantly lower than the values

[0043] Example 5: Measurement of changes in expression of cell death markers by auranofin treatment To measure changes in the expression of cell death markers due to auranofin treatment, RT4 and SC4 cell lines cultured as described in Example 1 were treated with auranofin at concentrations of 0, 0.1, and 0.3 μM, respectively, and 24 hours later, the protein expression of the cell death markers cleaved PARP and cleaved caspase-3 was compared by Western blot.

[0044] As shown in Figure 4, when RT4 and SC4 cell lines were treated with auranofin, the expression of cleaved PARP and cleaved caspase-3 increased in a concentration-dependent manner.

[0045] Example 6: Determination of the tumor growth suppression ability of auranofin in tumor-induced mice As shown in Figure 5A, 6-week-old female Balb / c nude mice with an average body weight of approximately 18 g were divided into three groups based on the dose of auranofin administered, and the SC4 cell line was injected into the flank of the mice to induce tumor formation.

[0046] For the low-concentration (2 mg / kg) auranofin, 0.04 mg of powdered auranofin was diluted in a mixture of 20 μL of DMSO (10%), 40 μL of PEG400 (20%), and 140 μL of PBS (200 μL total volume injected per 20 g mouse). For the high-concentration (10 mg / kg) auranofin, 0.2 mg of powdered auranofin was diluted in a mixture of 20 μL of DMSO (10%), 40 μL of PEG400 (20%), and 140 μL of PBS (200 μL total volume injected per 20 g mouse). Auranofin was intraperitoneally injected at 2 mg / kg or 10 mg / kg three times a week for two weeks. Control groups were injected with only the mixture of DMSO (10%), PEG400 (20%), and PBS. Tumor size was compared, as shown in Table 3. [Table 3] As shown in Figures 5B and 5C and Table 3, tumor growth was significantly suppressed in the auranofin-treated mice compared to the control group. In particular, the tumor growth rate in the experimental group administered auranofin at 10 mg / kg for 2 weeks was approximately 42.5% compared to the control group, confirming that tumor growth was effectively suppressed.

Claims

1. A pharmaceutical composition for preventing or treating neoplastic disorders, comprising auranofin as an active ingredient.

2. 2. The pharmaceutical composition for preventing or treating tumor diseases according to claim 1, wherein the tumor disease is at least one selected from the group consisting of schwannoma and neurofibromatosis type 2 syndrome.

3. The pharmaceutical composition for preventing or treating tumor diseases according to claim 2, wherein the schwannoma is vestibular schwannoma.

4. The pharmaceutical composition for preventing or treating tumor diseases according to claim 1, wherein the pharmaceutical composition contains auranofin at a concentration of 0.1 to 5 mg / mL.

5. 2. The pharmaceutical composition for preventing or treating tumor diseases according to claim 1, wherein the auranofin is administered in a dose of 1 to 20 mg per kg of body weight of the subject.

6. The pharmaceutical composition for preventing or treating tumor diseases according to claim 1, wherein the pharmaceutical composition is administered 2 to 5 times a week.

7. A method for preventing or treating a tumor disease, comprising administering to a non-human subject a composition comprising auranofin.

8. The method for preventing or treating a tumor disease according to claim 7, wherein the tumor disease is one or more selected from the group consisting of schwannoma and neurofibromatosis type 2.

9. The method for preventing or treating a tumor disease according to claim 8, wherein the neurinoma is acoustic neurinoma.

10. The method for preventing or treating a tumor disease according to claim 7, wherein the pharmaceutical composition contains auranofin at a concentration of 0.1 to 5 mg / mL.

11. The method for preventing or treating a tumor disease according to claim 7, wherein the auranofin is administered in a dose of 1 to 20 mg per kg of body weight of the subject.

12. The method for preventing or treating a tumor disease according to claim 7, wherein the composition containing auranofin is administered two to five times a week.

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