WO2020106049A1 - Pharmaceutical composition comprising both streptonigrin and anti-cancer agent for preventing or treating cancer - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or an adjuvant anti-cancer agent comprising both streptonigrin and gemcitabine for preventing or treating cancer. Additionally, the present invention relates to a method for preventing or treating cancer comprising the step for administering the pharmaceutical composition comprising both streptonigrin and gemcitabine to a patient, Therefore, the composition according to the present invention comprising both streptonigrin and other anti-cancer agents is highly effective in inhibiting cancer cell growth, especially the growth of pancreatic cancer cells, and thus can be effectively used in a composition or method using the composition for preventing or treating cancer.

Description

Pharmaceutical composition for use in the prevention or treatment of cancer, including both streptonigreen and anticancer agents

This application claims priority to Korean Patent Application No. 10-2018-0143938 filed on November 20, 2018, and the entire specification is a reference of the present application.

The present invention relates to pharmaceutical compositions or anticancer adjuvants for use in the prevention or treatment of cancer, including both streptonigrin and gemcitabine.

In addition, the present invention relates to a method for preventing or treating cancer comprising administering to a patient a pharmaceutical composition comprising both streptonigreen and gemcitabine.

Normal cells can perform regular and elastic proliferation and suppression as needed, whereas cancer cells proliferate indefinitely, which is also called a tumor as a cell mass composed of undifferentiated cells. These cancer cells penetrate into surrounding tissues and metastasize to other organs of the body, causing severe pain and eventually death. Despite the development of medicine, the number of cancer patients in Korea has increased continuously, increasing by about 44% in the last 10 years, and the market for anticancer drugs has also increased internationally, and has been reported to have a size of about 100 billion dollars per year.

Anticancer drugs are chemotherapy drugs, which are first-generation anti-cancer drugs, and targeted anti-cancer drugs, which are second-generation anti-cancer drugs. To overcome these side effects, immuno-cancer drugs have been developed as third-generation anti-cancer drugs and research is ongoing. However, the biggest problem in the treatment of cancer is the recurrence of cancer, which is why it is difficult to target a specific cancer due to various mutations in cancer, and resistance to anticancer drugs used in the treatment of recurrent cancer occurs. This is because the case is non-existent. As a result, even after treatment of primary cancer, most patients die from metastatic and recurrent cancer. Accordingly, in order to enhance the effectiveness of the anti-cancer agent, a strategy to combine and treat the anti-cancer agent has been proposed.

Transglutaminase (Transglutaminase 2, TGase2) is an enzyme that promotes the binding between γ-carboxamide groups of glutamine residues bound to specific peptides and various amines, and primarily promotes the prevention, repair, and repair of damage. Although it is known to play a major role in recent studies, recent studies have shown that abnormally excessive expression may contribute to the development of diseases such as neurodegenerative diseases, atherosclerosis, inflammatory diseases, and autoimmune diseases. There is a bar. In particular, it has been reported that the expression of TGase2 polymerizes and destabilizes p53 and thus disappears. Accordingly, it has been reported that the inhibition of TGase2 may have an anti-cancer effect on overexpressed kidney cancer.

Republic of Korea Patent Publication No. 10-2016-0009146 provides a pharmaceutical composition for treating or preventing a disease caused by an increase in transglutaminase 2 activity including streptonigreen, pancreatic cancer, uterine cancer to the disease It is disclosed that breast cancer is included.

However, studies or descriptions have not been disclosed to generate a synergistic effect on the anti-cancer effect, such as suppressing the growth of cancer cells by using streptonigreen and other anti-cancer agents in combination.

Accordingly, the present inventors tried to provide a composition having a significantly elevated anti-cancer effect by using streptonigrin and other anti-cancer agents, and as a result, superior anti-cancer effect when treated with cancer cells, particularly pancreatic cancer cells, using streptonigreen and gemcitabine in combination It confirmed that it represents and completed this invention.

Accordingly, an object of the present invention is to provide a pharmaceutical composition, an anticancer adjuvant to a method for preventing or treating cancer, for use in preventing or treating cancer comprising both streptonigrin and gemcitabine.

In order to achieve the above object, the present invention provides a pharmaceutical composition or an anticancer adjuvant for use in the prevention or treatment of cancer, including both streptonigrin and gemcitabine.

In one preferred embodiment of the present invention, the streptonigreen and gemcitabine may be included in a concentration ratio of 1: 1 to 1: 10000.

In a preferred embodiment of the present invention, the cancer may be pancreatic cancer.

In addition, the present invention provides a method for preventing or treating cancer, comprising administering to a patient a pharmaceutical composition comprising both streptonigrin and gemcitabine.

In one preferred embodiment of the present invention, the streptonigreen and gemcitabine may be included in a concentration ratio of 1: 1 to 1: 10000.

In one preferred embodiment of the present invention, the cancer may be pancreatic cancer.

As described above, in the prior art, it was confirmed that the anti-cancer effect can be exhibited by using the transglutaminase (TGase2) inhibitory effect of streptonigreen, and when used in combination with the anti-cancer agent, the anticancer effect is significantly increased. No studies or descriptions of synergistic effects have been disclosed.

Since the composition comprising both streptonigreen and other anticancer agents according to the present invention inhibits the growth or growth of cancer, it generates a significant anticancer effect, and thus can be effectively used in cancer prevention or treatment compositions, anticancer adjuvants or cancer prevention or treatment Can be.

Accordingly, the present invention can provide a pharmaceutical composition for use in the prevention or treatment of cancer, which includes both streptonigrin and gemcitabine.

The Streptonigreen is a Tgase2 inhibitor produced by Streptomyces floculus, and refers to an antibiotic that causes chromosomal cleavage. The gemcitabine is an anticancer agent used for various cancers such as breast cancer, ovarian cancer, or pancreatic cancer, and has an effect of inhibiting DNA production related to cell death.

The streptonigreen and gemcitabine are preferably included in a concentration ratio of 1: 1 to 1: 10000, more preferably, in a concentration ratio of 1:10 to 1: 1000, most preferably 1: It is preferably included in a concentration ratio of 50 to 1: 300.

It is preferred that the cancer of the present invention is pancreatic cancer.

The pharmaceutical composition for use in the prevention or treatment of cancer of the present invention may be various oral or parenteral formulations. When formulating the composition, one or more buffers (e.g. saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g. EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g. Aluminum hydroxide), suspending agents, thickener wetting agents, disintegrating agents or surfactants, diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in one or more compounds, such as starch (corn starch, wheat starch, rice starch, potato Starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl -It is prepared by mixing cellulose or gelatin. For example, tablets or dragees can be obtained by blending the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granular mixture.

In addition, lubricants such as magnesium stearate, talc and the like are used in addition to simple excipients. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions or syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, flavoring agents or preservatives, etc. are included. Can be. In addition, if necessary, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and may further include an anti-coagulant, lubricant, wetting agent, fragrance, emulsifier, and preservative. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspension solutions, emulsions, lyophilized preparations or suppositories. As the non-aqueous solvent and suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, etc. may be used.

The composition of the present invention may be administered orally or parenterally, and for parenteral administration, external application to the skin; Injections that are injected intraperitoneally, rectal, intravenous, intramuscular, subcutaneous, intrauterine dura mater or cerebrovascular; Transdermal administration; Alternatively, nasal inhalants may be formulated according to methods known in the art.

The injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils. Can be. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with triethanol amine or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. may be further included. In addition, the injection may additionally contain isotonic agents such as sugars or sodium chloride in most cases.

In the case of transdermal dosage forms, ointments, creams, lotions, gels, external solutions, pasta agents, linen agents, aerosols, and the like are included. In the above, transdermal administration means that the pharmaceutical composition is topically administered to the skin, so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

In the case of inhalation dosages, the compounds used according to the invention may be used in the form of pressurized packs, using suitable propellants, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a compound and a suitable powder base such as lactose or starch.

The pharmaceutical composition for use in the prevention or treatment of cancer of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the patient's disease type, severity, drug activity, sensitivity to the drug, and administration time. , The route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. That is, the total effective amount of the composition of the present invention can be administered to a patient in a single dose, and can be administered by a fractionated treatment protocol that is administered for a long time in multiple doses. . Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity. As a daily dose, when administered parenterally, it is preferably administered in an amount of 0.01 to 50 mg per kg of body weight per day, more preferably 0.1 to 30 mg per kg of body weight per day, based on streptonigreen and an anticancer agent used in combination, and orally When administered, it can be divided into 1 to several times to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per 1 kg of body weight per day, based on the streptonigrin of the present invention and an anticancer agent used in combination. have. However, since the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, and age, the above dosage does not limit the scope of the present invention in any way.

The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormonal therapy, chemotherapy and biological response modifiers.

The pharmaceutical composition of the present invention can also be provided as a formulation of an external preparation containing streptonigreen and an anticancer agent used in combination therewith as an active ingredient. When the pharmaceutical composition for use in the prevention or treatment of cancer of the present invention is used as an external preparation for skin, fat substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, blowing agents (foaming agent), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, metal ion blocker, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, It may contain adjuvants commonly used in the field of skin science, such as lipophilic actives or any other ingredients commonly used in external skin preparations such as lipid vesicles. In addition, the ingredients may be introduced in an amount commonly used in the field of skin science.

When the pharmaceutical composition for use in the prevention or treatment of cancer of the present invention is provided as an external preparation for skin, the present invention is not limited thereto, and may be a formulation such as ointment, patch, gel, cream or spray.

In addition, the present invention can provide an anti-cancer adjuvant comprising both streptonigrin and gemcitabine.

Since the streptonigreen and gemcitabine are the same as those used in the composition for preventing or treating cancer, the description is replaced by the above description.

The anti-cancer adjuvant of the present invention means all forms for enhancing the anti-cancer effect of the anti-cancer agent or suppressing or improving side effects of the anti-cancer agent. The anti-cancer adjuvant of the present invention may be administered in combination with various types of anti-cancer agents or anti-cancer adjuvants. Treatment can be performed.

 The route of administration of the anticancer adjuvant may be administered through any general route as long as it can reach the target tissue. The anti-cancer adjuvant of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, orally, pulmonarily, intrarectally, as desired, but is not limited thereto. In addition, the anti-cancer adjuvant may be administered by any device capable of transporting the active substance to the target cell.

 The anticancer adjuvant of the present invention can be preferably formulated as an anticancer adjuvant by including at least one pharmaceutically acceptable carrier in addition to the active ingredient for administration. Carriers, excipients, or diluents that may be included in the anticancer treatment adjuvant of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

 The anti-cancer adjuvant of the present invention may be an agent for oral or parenteral administration, and the description of the agent is replaced by the description of the agent of the pharmaceutical composition.

In addition, the present invention can provide a method for preventing or treating cancer comprising the step of administering to a patient a pharmaceutical composition comprising both streptonigreen and gemcitabine.

The streptonigreen, gemcitabine and pharmaceutical composition are the same as those used in the pharmaceutical composition for use in the prevention or treatment of cancer, so the description is replaced by the above description.

The administration is a concept including both administration of streptonigreen and gemcitabine simultaneously or sequentially.

It is preferred that the cancer is pancreatic cancer.

Therefore, the composition comprising both the streptonigreen of the present invention and other anticancer agents is effective as a composition for preventing or treating cancer because it has an excellent effect of inhibiting the growth of cancer cells, especially the effect of inhibiting the growth of pancreatic cancer cells.

Figure 1 shows the volume of the tumor when treated with pancreatic cancer tumors (MIA PaCa-2 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. Compared to the solvent or the alone treatment group, the tumor volume was most significantly reduced when streptonigreen and gemcitabine were used in combination.

Figure 2 shows the size of the tumor when treated with pancreatic cancer tumors (MIA PaCa-2 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine, or streptonigreen and gemcitabine. The tumor size was most significantly reduced when treated with streptonigrin and gemcitabine in combination with the solvent or alone treatment group.

Figure 3 shows the weight of the tumor when treated with pancreatic cancer tumor (MIA PaCa-2 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. Tumor weight was most significantly decreased when streptonigrin and gemcitabine were used in combination with a solvent or a single treatment group.

Figure 4 shows the weight of the rat when treated with pancreatic cancer tumor (MIA PaCa-2 cells) in combination with a solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. It was confirmed that the weight of the rat increased from 3 weeks after the combination treatment when treated with streptonigrin and gemcitabine in combination with the solvent or the alone treatment group.

Figure 5 shows the volume of the tumor when treated with pancreatic cancer tumors (BxPC-3 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigrin, gemcitabine, or streptonigrin and gemcitabine. Compared to the solvent or the alone treatment group, the tumor volume was most significantly reduced when streptonigreen and gemcitabine were used in combination.

Figure 6 shows the size of the tumor when treated with pancreatic cancer tumors (BxPC-3 cells) in combination with a solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. The tumor size was most significantly reduced when treated with streptonigrin and gemcitabine in combination with the solvent or alone treatment group.

Figure 7 shows the weight of the tumor when treated with pancreatic cancer tumors (BxPC-3 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. The weight of the tumor was most significantly reduced when treated with streptonigrin and gemcitabine in combination with the solvent or alone treatment group.

Figure 8 shows the weight of the rat when treated with pancreatic cancer tumors (BxPC-3 cells) in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. It was confirmed that the weight of the rat increased from 3 weeks after the combination treatment when treated with streptonigrin and gemcitabine in combination with the solvent or the alone treatment group.

Figure 9 shows the growth of cells when treated with pancreatic cancer cells in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, gemcitabine or streptonigreen and gemcitabine. It was confirmed that the growth rate of pancreatic cancer cells was significantly reduced when streptonigrin and gemcitabine were used in combination compared to a solvent or a single treatment group.

Figure 10 shows the growth of cells when treated with pancreatic cancer cells in combination with solvent (DMSO 1%, PBS 99%), streptonigreen, paclitaxel or screptonigreen and paclitaxel. It was confirmed that the synergistic effect by the combination treatment did not appear.

[Example 1]

Preclinical xenograft tumor model

A preclinical xenograft tumor model was prepared to confirm the effect of streptonigreen and / or gemcitabine on pancreatic cancer tumors.

Specifically, tumors were induced in Balb / c-nu mice (Central Lab Animal, Seoul, Korea) of 6 to 8 weeks of age. MiaPaCa-2 cells or BxPC-3 cells (1.0 × 10 ⁷ ) were inoculated subcutaneously using a 1 ml syringe. After 2 weeks, mice were treated with solvent (DMSO 1%, PBS 99%), streptonigreen (KN383), gemcitabine or streptonigreen and gemcitabine (streptonigreen 0.1 mg / kg / 100 μl (PO), gemcitabine 100mg / kg / 100µl (IP), 6 animals in each group).

As a result, as shown in [FIG. 1] to [FIG. 8], when streptonigreen and gemcitabine are used in combination, the volume of the tumor does not increase and the weight of the tumor is significant as compared to the treatment with each alone. It was confirmed that the decrease and the weight of rats was confirmed to increase from 3 weeks after drug treatment.

[Example 2]

Cell culture

Pancreatic cancer cells MIA PaCa-2, BxPC-3, Capan2, AsPC-1 and SNU-324 cells were received in the laboratory of ph.D Heo Kyun. The pancreatic cancer cells were cultured in a complete RPMI 1640 medium (Hyclone, UT, USA) containing 10% fetal bovine serum (Hyclone, UT, USA) in an environment of 5% CO _2, 100% humidity and 37 ° C.

[Example 3]

Cancer cell proliferation analysis

In the case of treatment with streptonigrin and / or anticancer agent, the change in cancer cell proliferation was attempted to be confirmed through a sulforhodamine B assay.

Specifically, cells (100 μl) were seeded in 96-well microtiter plates with plating densities of 5000 to 40000 cells / well depending on the doubling time of individual cell lines. After cell inoculation, the cells were cultured for 24 hours prior to the addition of the experimental drug. After 100 μl of each drug was added to each well, the plate was incubated for 48 hours in a 37 ° C. CO ₂ incubator. Cells were treated with cold 50% TCA (trichloroacetic acid, 50 μl / well), and then incubated at 4 ° C. for 60 minutes to fix the cells. The supernatant was discarded, washed 5 times with tap water and naturally dried at room temperature for 12 to 24 hours. A 0.4% (w / v) solution of sulforhodamine B (100 μl) in 1% acetic acid was added to each well, and the plate was left at room temperature for 10 minutes. After staining, washing was performed 5 times with 1% acetic acid to remove non-binding dyes and allowed to air dry. The bound dye was then solubilized with 10 mM Trizma base and absorbance was recorded using an automated plate reader at 515 nm. The effect of the drug was expressed as GI50 (50% growth inhibition), TGI (total growth inhibition) or LC50 (lethal concentration).

In the case of pancreatic cancer, the concentration of each of the treated drugs is 20 nM or 50 nM for streptonigreen, 5 μM for gemcitabine, and 10 nM for paclitaxel.

As a result, in the case of pancreatic cancer cells, as shown in [Fig. 9], when streptogrin and gemcitabine were used in combination, it was confirmed that the cell growth inhibitory effect was significantly increased compared to the treatment with each alone. . On the other hand, when treated in combination with streptonigreen and paclitaxel, as shown in [Fig. 10], it was confirmed that synergistic effect does not occur compared to the treatment with each alone.

The composition comprising both streptonigreen and gemcitabine provided by the present invention has a synergistic effect and is excellent in inhibiting cancer, particularly pancreatic cancer, so that it is a pharmaceutical composition for use in preventing or treating cancer, an anticancer adjuvant or using the same Since it can be effectively used for cancer prevention or treatment, it has great industrial applicability.

Claims (8)

1. A pharmaceutical composition for use in the prevention or treatment of cancer, comprising both streptonigreen and gemcitabine.
2. The method of claim 1, wherein the streptonigreen and gemcitabine is a pharmaceutical composition for use in the prevention or treatment of cancer, characterized in that it is included in a concentration ratio of 1: 1 to 1: 10000.
3. The pharmaceutical composition for use in the prevention or treatment of cancer according to claim 1, wherein the cancer is pancreatic cancer.
4. An anti-cancer adjuvant containing both streptonigreen and gemcitabine.
5. The anticancer adjuvant according to claim 4, wherein the streptonigreen and gemcitabine are contained in a concentration ratio of 1: 1 to 1: 10000.
6. A method of preventing or treating cancer, comprising administering to a patient a pharmaceutical composition comprising both streptonigreen and gemcitabine.
7. The method for preventing or treating cancer according to claim 6, wherein the streptonigreen and gemcitabine are contained in a concentration ratio of 1: 1 to 1: 10000.
8. The method for preventing or treating cancer according to claim 6, wherein the cancer is pancreatic cancer.

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