KR20200096141A - Composition for preventing or treating colon cancer comprising streptonigrin and anticancer agent - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or anti-cancer aid for preventing or treating colorectal cancer, including streptonigrin and an anti-cancer agent. The present invention also relates to a pharmaceutical composition or anti-cancer aid for preventing 5-FU resistant colorectal cancer, including streptonigrin. The composition including streptonigrin and oxaliplatin, or streptonigrin and 5-FU according to the present invention shows a significantly increased effect of inhibiting growth of colorectal cancer cell strains, as compared to the treatment using the anti-cancer agent alone, and thus can be provided as a composition for preventing or treating colorectal cancer. In addition, in the case of 5-FU resistant colorectal cancer cell strains, even though treatment with streptonigrin alone shows an anti-cancer activity, treatment using streptonigrin and 5-FU in combination shows a synergic effect to provide a significantly increased anti-cancer activity.

Description

A pharmaceutical composition for preventing or treating colon cancer comprising streptonigrin and an anticancer agent {Composition for preventing or treating colon cancer comprising streptonigrin and anticancer agent}

The present invention relates to a pharmaceutical composition or an anticancer adjuvant for preventing or treating colon cancer comprising streptonigrin and an anticancer agent.

In addition, the present invention relates to a pharmaceutical composition or anticancer adjuvant for preventing or treating 5-FU-resistant colorectal cancer comprising streptonigrin.

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 serious pain and eventually death. Despite the advancement of medicine, the number of cancer patients in Korea has continuously increased, increasing by about 44% in the last 10 years, and the anticancer drug market has also increased internationally, which is reported to have a scale 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, most patients die from metastases and recurring cancers even after treatment of primary cancer. Accordingly, in order to enhance the effectiveness of the anti-cancer agent, a strategy to mix 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. It is primarily known to play a major role in the prevention of damage, and in promoting defense and repair, but recent studies have shown that abnormally overexpressed neurodegenerative diseases, atherosclerosis, inflammatory diseases, and autoimmune diseases It has been reported that it can provide a cause for disease outbreaks. In particular, it has been reported that the expression of TGase2 polymerizes and destabilizes p53, and thus it has been reported that TGase2 can exhibit an anticancer effect against overexpressed kidney cancer through inhibition of TGase2.

Republic of Korea Patent Laid-Open No. 10-2016-0009146 provides a pharmaceutical composition for treating or preventing diseases caused by increased transglutaminase 2 activity including streptonigrin, and pancreatic cancer, uterine cancer or It is disclosed that breast cancer is included.

However, no research or description has been disclosed to generate a synergistic effect on the anticancer effect, such as inhibition of the growth of cancer cells by using streptonigrin and other anticancer agents in combination.

Accordingly, the present inventors have used streptonigreen and oxaliplatin or 5-FU, respectively, to treat and treat cancer cell lines, particularly colon cancer cell lines, as a result of diligent efforts to research and provide substances capable of significantly increasing anti-cancer activity when used in combination. It was confirmed that the case can show a significantly increased anticancer activity compared to the treatment alone, and the present invention was completed.

Accordingly, an object of the present invention is to provide a pharmaceutical composition or anticancer adjuvant for preventing or treating colon cancer, including streptonigrin and an anticancer agent.

Another object of the present invention is to provide a pharmaceutical composition or anticancer adjuvant for preventing or treating 5-FU-resistant colorectal cancer comprising streptonigrin.

In order to achieve the above object, the present invention provides a pharmaceutical composition or anticancer adjuvant for preventing or treating colon cancer comprising streptonigrin and an anticancer agent.

According to a preferred embodiment of the present invention, the anti-cancer agent may be oxaliplatin or 5-FU.

According to a preferred embodiment of the present invention, the streptonigreen and anticancer agent may be included in a molar concentration ratio of 1: 1 to 1: 10 ⁵ .

The present invention also provides a pharmaceutical composition or an anticancer adjuvant for preventing or treating 5-FU-resistant colorectal cancer, including streptonigrin.

According to a preferred embodiment of the present invention, the composition may further include 5-FU.

According to a preferred embodiment of the present invention, the Streptomyces you green and 5-FU is from 1: may be included at a molar concentration ratio of 10 ^6: 1 to 1.

According to one preferred embodiment of the present invention, the 5-FU-resistant colon cancer may be over-expressing transglutaminase 2 (Transglutaminase 2).

Therefore, the composition of the present invention contains streptonigrin and oxaliplatin or streptonigrin and 5-FU, so that the effect of inhibiting the growth of colon cancer cell lines is significantly increased compared to the case of treatment alone. Or it may be provided as a therapeutic composition. In addition, in the case of 5-FU-resistant colorectal cancer cell lines, anti-cancer activity is shown even when streptonigrin is treated alone, but synergism occurs when streptonigrin and 5-FU are used in combination, indicating a significantly increased anti-cancer activity. have.

1 is a treatment by using streptonigreen and oxaliplatin or streptonigreen and 5-FU in combination, respectively, compared to the case where streptonigrin (STN), oxaliplatin, or 5-FU is treated alone in colon cancer cell lines In the case of (combi), it indicates that the anticancer activity against colorectal cancer cell lines is effectively increased. In particular, it was confirmed that when streptonigrin was used in combination with oxaliplatin, it was effective against HCT-116 and DLD1 cell lines, and when streptonigrin was used in combination with 5-FU, it was confirmed that it effectively exhibited anticancer activity against HCT-116 cell line.
Figure 2 shows that the synergistic effect on anticancer activity did not occur even if streptonigrin (STN) and cisplatin or streptonigrin and paclitaxel were treated in combination with each of the colon cancer cell lines. It also indicates that the anticancer activity is lower than that of the case.
Figure 3 shows the expression level of transglutaminase 2 (TGase 2) in the 5-FU-resistant colon cancer cell line (FR) and wild-type colon cancer cell line (HCT-15, WT). It was confirmed that the expression of TGase 2 was significantly increased in FR compared to WT.
Figure 4 shows the survival rate of 5-FU-resistant colorectal cancer cell lines (Resistance) and wild-type colorectal cancer cell lines (WT) treated with 5-FU by concentration. When treated with 1 μM of 5-FU, compared to WT whose survival rate begins to decrease, in the case of a 5-FU-resistant colon cancer cell line, treatment with 100 μM of 5-FU did not affect the cell survival rate.
5 shows that the anticancer activity significantly increased when the 5-FU-resistant colorectal cancer cell line (HCT-15FR) was treated with streptonigrin or 5-FU alone compared to the case of treatment alone.
Figure 6 shows the volume change when treated with streptonigrin and/or 5-FU to a colon cancer cell line (HCT116). It was confirmed that the tumor volume was significantly decreased when streptonigrin and 5-FU were treated in combination compared to the case of treatment with streptonigrin or 5-FU alone.
Figure 7 shows the weight change when treated with streptonigrin and/or 5-FU to the colon cancer cell line (HCT116). It was confirmed that the weight of the tumor was significantly reduced when streptonigrin and 5-FU were treated in combination compared to the case of treatment with streptonigrin or 5-FU alone.

Hereinafter, the present invention will be described in more detail.

As described above, in the prior art, there was a problem that resistance to an anti-cancer agent using an anti-cancer agent against cancer patients may occur, and the anti-cancer effect using the effect of inhibiting transglutaminase (TGase2) of streptonigreen was used. Although it has been confirmed that it can be exhibited, studies or descriptions of synergistic effects with anticancer agents that significantly increase the anticancer effect when used in combination with it have not been disclosed.

Since the composition comprising streptonigrin and other anticancer agents according to the present invention significantly inhibits the growth or growth of cancer cell lines, especially colon cancer-related cell lines, it exhibits a synergistic effect on anticancer activity, and thus, a pharmaceutical for preventing or treating colon cancer It is effective as a composition or anticancer adjuvant.

Streptonigrin of the present invention is a Tgase2 inhibitor produced by Streptomyces floculus, and refers to an antibiotic that causes chromosomal cleavage.

Oxaliplatin of the present invention is a vaccine-based antitumor agent-based drug, and is known to have an effect of inhibiting DNA replication of cancer cells.

5-FU of the present invention is also called fluorouracil, and is known to inhibit DNA production by inhibiting thymidylate synthase.

When treating a composition comprising a combination of strepttonirin and oxaliplatin in a colorectal cancer cell line, or a combination of strepttonirin and 5-FU, it is significant compared to treatment with strepttonirin, oxaliplatin or 5-FU, respectively. As a result, an effect of inhibiting survival or growth of the colon cancer cell line may be increased ([ FIG. 1 ], [FIG. 6 ], and [FIG. 7 ]). However, streptonigreen is not capable of generating synergistic effects against all anticancer drugs. In the case of combination of streptonirin and cisplatin or streptonirin and paclitaxel, synergistic effects did not occur on anticancer activity, but rather anticancer activity may be reduced compared to the single treatment group (FIG. 2).

When treatment with streptonigreen alone in an anticancer agent, particularly a colon cancer cell line resistant to 5-FU, can effectively suppress its survival. However, when the cell line is treated with streptonigreen and 5-FU in combination, a synergistic effect is generated, and thus a more elevated cell line survival inhibitory effect can be obtained (FIG. 5). This can be presumed to be due to the overexpression of transglutaminase 2 in the case of colorectal cancer cell lines resistant to 5-FU (FIG. 4).

Accordingly, the present invention can provide a pharmaceutical composition or anticancer adjuvant for preventing or treating colon cancer, including streptonigrin and an anticancer agent.

The anticancer agent of the present invention is preferably oxaliplatin or 5-FU. That is, the pharmaceutical composition or anticancer adjuvant for preventing or treating colon cancer may contain streptonigrin and oxaliplatin, or may include streptonigrin and 5-FU. The anticancer adjuvant may be effective when streptonigrin and an anticancer agent are administered simultaneously or sequentially.

The streptonigreen and anti-cancer agent of the present invention is preferably included in a molar concentration ratio of 1: 1 to 1: 10 ⁵ , more preferably 1: 10 to 1: 10 ⁴ may be included in a molar concentration ratio, most preferably It is most preferably included in a molar concentration ratio of 1: 30 to 1: 10 ³ .

In addition, the present invention can provide a pharmaceutical composition or anticancer adjuvant for preventing or treating 5-FU-resistant colon cancer, including streptonigrin.

It is preferable to additionally include 5-FU in the pharmaceutical composition to the anticancer adjuvant, in which case streptonigrin and 5-FU are preferably included in a molar ratio of 1:1 to 1:10 ⁶ . More preferably, it is preferably included in a molar concentration ratio of 1: 10 to 1: 10 ⁵ , most preferably it is included in a molar concentration ratio of 1: 100 to 1: 10 ⁴ . When additionally including 5-FU in the anticancer adjuvant, it may be effective that streptonigrin and 5-FU are administered simultaneously or sequentially.

The 5-FU-resistant colorectal cancer of the present invention may be to overexpress Transglutaminase 2 (Transglutaminase 2).

The pharmaceutical composition for preventing or treating colon cancer of the present invention may be oral or parenteral in various dosage forms. When formulating the composition, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., Aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants 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, pulverizing it, adding a suitable auxiliary, and processing into a granule 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, are included. Can. In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and an anti-coagulant, a lubricant, a wetting agent, a fragrance, an emulsifier and a preservative, etc. may be additionally included. .

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 pharmaceutical composition of the present invention may be administered orally or parenterally, and when applied parenterally, externally applied 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. Examples of suitable carriers for injections include, but are not limited to, water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), mixtures thereof and/or solvents or vegetable media comprising vegetable oils Can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with ethanol 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, it may further include various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, the injection may additionally include an isotonic agent such as sugar or sodium chloride in most cases.

In the case of transdermal dosage forms, ointments, creams, lotions, gels, external solutions, pasta, 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 can be used in a pressurized pack or with a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. It can be conveniently delivered from a nebulizer in the form of an aerosol spray. 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 preventing or treating colon 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, drug sensitivity, 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 conventional therapeutic agents, and may be administered single or multiple. That is, the total effective amount of the composition of the present invention may be administered to a patient in a single dose, and may be administered by a fractionated treatment protocol that is administered for a long period of 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 streptonigreen of the present invention and the anticancer agent used in combination with the present invention. have. However, since the dosage may be increased or decreased depending on the route of administration, the severity of obesity, sex, weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

The composition of the present invention can 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 may also be provided in the form of an external preparation comprising streptonigrin and an anticancer agent used in combination therewith as an active ingredient. When the pharmaceutical composition for preventing and treating colorectal 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, foaming agents ), 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, lipophilic active agent Or it may contain an adjuvant commonly used in the field of skin science, such as any other ingredients commonly used in external preparations for skin such as lipid vesicles. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

When the pharmaceutical composition for preventing and treating colon 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.

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 administration route of the anticancer adjuvant may be administered through any general route as long as it can reach the target tissue. The anticancer adjuvant of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, orally, pulmonarily, or rectally, 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 adjuvants 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 anticancer 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.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Colon cancer cell line culture

The COLO205, HCT-116, KM-12, HCT-15, HCT-8, SW-620 and HT29 cell lines were obtained from the National Cancer Institute (MTA: 2702-09), and the DLD1 cell line was obtained from a Korean cell line bank. . Cells were cultured in complete RPMI 1640 medium (Hyclone, UT, USA) containing 10% fetal bovine serum (Hyclone, UT, USA) in an environment of 37° C., 5% CO ₂ and 100% humidity, respectively.

The 5-FU resistant colorectal cancer cell line HCT-15-FR was prepared by exposing the HCT-15 cell line to a 5-FU concentration gradient. Specifically, HCT-15 cells were cultured in RPMI 1640 medium without drugs for 24 hours, and then, the medium was replaced with complete medium to which 5 μM 5-FU was added and cultured for 72 hours. When the cells grow to 80% (confluence), the above steps were repeated several times. As a result, HCT-15 cells cultured until stable growth at 100 μM of 5-FU, that is, 5-FU resistant cell line HCT15-FR was obtained.

SRB assay for antiproliferative activity

Cells (100 μl) were seeded into 96-well microtiter plates at a plating density of 5000 to 40000 cells/well depending on the doubling time of individual cell lines. After incubation for 24 hours before drug addition, an anticancer agent was added to each well, and the culture solution was incubated at 37°C for 48 hours. Cells were fixed in 50% trichloroacetic acid (50 μl/well), and then incubated at 4° C. for 1 hour. The liquid was removed from the plate, then rinsed 5 times with water and dried at room temperature for about 12-24 hours. After staining the fixed cells with 100 μl SRB for 5 minutes at room temperature, the plate was washed 3 times with 1% glacial acetic acid and dried at room temperature for about 12 to 24 hours. The SRB stain was then solubilized in 10 mM Trizma base and absorbance read at 515 nm. The effect of the drug was expressed as GI50 (50% growth inhibition), TGI (total growth inhibition) or LC50 (lethal concentration).

Western blotting

Total cell lysates of wild-type HCT-15 cell line (WT) or 5-FU resistant colorectal cancer cell line HCT15-FR(FR) include 150 mM sodium chloride, 1.0% IGEPAL CA-630 (NP-40), in addition to 50 mM Tris-HCl. Prepared using RIPA buffer (pH 8.0) containing a mixture of 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, protease inhibitor and phosphatase inhibitor. Protein analysis was performed using BCA protein analysis (Thermo Fisher, IL, USA) to normalize the protein. Proteins were analyzed by SDS-PAGE and transferred to a Polyvinylidene difluoride (PVDF) membrane (Merck Millipore, MA, USA). Membranes were blocked in 5% BSA for 1 hour at room temperature and incubated at 4° C. overnight with the indicated antibodies. The membrane was then washed with TBS-T for 1 hour at room temperature, and then incubated with horseradish peroxidase-binding secondary antibody for 1 hour at room temperature. After incubation, the membrane was washed with TBS-T for 1 hour at room temperature to confirm by enhanced chemiluminescence (ECL).

As a result, as shown in [Fig. 3], it was confirmed that the expression level of transglutaminase 2 (TGase 2) was significantly increased in FR compared to WT.

Confirmation of the combination effect of streptonigreen and anticancer drug

In the case of using a combination of streptonigreen and an anticancer agent (oxaliplatin or cisplatin) in a wild-type colon cancer cell line, that is, colon cancer cell lines that are not resistant to 5-FU, it was attempted to confirm whether synergy occurs in anticancer activity.

Specifically, the colon cancer cell lines COLO205, HCT-116, KM-12, HCT-15, HCT-8, SW-620, DLD1 and HT29 have streptonigreen (STN) of 20 nM, oxaliplatin of 1 μM, cisplatin ( cisplatin) was treated with 20 μM, paclitaxel with 20 nM and 5-FU with 5 μM, respectively, or in combination, and the survival rate of the colorectal cancer cell line was confirmed by the method of <Example 2>.

As a result, as shown in FIG. 1, streptonigrin and oxaliplatin or streptonigrin and 5-FU were used in combination, compared to the case where streptonigrin, oxaliplatin, or 5-FU was treated alone in colon cancer cell lines. In the case of treatment (combi), it was confirmed that the anti-cancer activity against colon cancer cell lines was effectively increased. In particular, it was confirmed that when streptonigrin was used in combination with oxaliplatin, it was effective against HCT-116 and DLD1 cell lines, and when streptonigrin was used in combination with 5-FU, it was confirmed that it effectively exhibited anticancer activity against HCT-116 cell line.

However, as shown in [Fig. 2], when treating streptonigrin and cisplatin or streptonirin and paclitaxel in combination with colon cancer cell lines, synergistic effect on anticancer activity did not occur, but rather each was treated alone. Compared to the case, it was confirmed that the occurrence of the anticancer activity was also lowered.

Confirmation of the combined effect of streptonigreen and 5-FU

In the case of treating the 5-FU-resistant colon cancer cell line prepared in the present invention with streptonigreen and 5-FU in combination, it was intended to confirm whether synergy occurs in anticancer activity.

Specifically, the 5-FU-resistant colon cancer cell line (Resistance) prepared in <Example 1> and wild-type colon cancer cell line (WT) by 5-FU concentration (1nM, 10nM, 100nM, 1μM, 10μM and 100μM) After treatment with the cell was confirmed the survival rate. In addition, the 5-FU-resistant colorectal cancer cell line (HCT-15FR) and wild-type colorectal cancer cell line (WT) prepared in <Example 1> were treated with 20 nM of streptonigrin and 50 μM of 5-FU, respectively or in combination. After that, the survival rate of the colon cancer cell line was confirmed by the method of <Example 2>.

As a result, as shown in [Fig. 4], compared to WT, where the survival rate begins to decrease when 1 μM of 5-FU is treated, the cells of the 5-FU-resistant colon cancer cell line are treated with 100 μM of 5-FU. It was confirmed that there was no effect on the survival rate.

In addition, as shown in [Fig. 5], 5-FU resistant colorectal cancer cell line (HCT-15FR) is treated with streptonigrin or 5-FU, respectively, when treated in combination (combination), anticancer activity is significant. It was confirmed that the enemy increased.

Preclinical xenograft tumor model

The experiments of the present invention were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the National Cancer Center Institute accredited by the American National Cancer Center Institute (AAALAC International) following the Institute of Laboratory Animals Research. . HCT-116 cells (5×10 ^{6 in} 100 μl PBS) in Balb/c-nu mice (Orient, Seoul, Korea) of 6 to 8 weeks of age Was inoculated subcutaneously with a 1-mL syringe. After a week, mice were divided into 4 groups: solvent control (Control), streptonigrin, 5-FU, and 5-FU combination with streptonigrin (Streptonigrin + 5-FU, Combination). Streptonigreen (0.1 mg/kg body weight), 5-FU (40 mg/kg body weight) and solvent (1% DMSO + 99% PBS) orally 5 days/week (Streptonigreen), or 3 days intraperitoneally /Week (5-FU). The primary tumor size was measured weekly using a caliper. Tumor volume was calculated as V = (A × B ² )/2 (V: volume (mm ³ ), A: long diameter, B: short diameter).

As a result, as shown in [Fig. 6] and [Fig. 7], the volume and weight of the tumor when treated with Streptonigreen and 5-FU in combination compared to the case where Streptonigreen or 5-FU are treated alone, respectively. Was found to decrease significantly.

Claims (14)

A pharmaceutical composition for preventing or treating colon cancer, including streptonigrin and an anticancer agent, or for preventing or treating colon cancer, including streptonigrin and an anticancer agent.
[2] The pharmaceutical composition for preventing or treating colon cancer according to claim 1, wherein the anticancer agent is oxaliplatin or 5-FU.
The method of claim 1 wherein the Streptomyces you green and anticancer agents is from 1: 1 to 1: colon, characterized in that contained as the molar concentration ratio of 10 ⁵ or therapeutic pharmaceutical composition for preventing cancer.
5-FU-resistant colorectal cancer prevention or treatment pharmaceutical composition comprising streptonigrin (streptonigrin).
The pharmaceutical composition for preventing or treating 5-FU-resistant colon cancer according to claim 4, wherein the composition additionally contains 5-FU.
The pharmaceutical composition for preventing or treating 5-FU-resistant colon cancer according to claim 5, wherein the streptonigrin and 5-FU are contained in a molar ratio of 1: 1 to 1: 10 ⁶ .
The pharmaceutical composition for preventing or treating 5-FU-resistant colorectal cancer according to claim 4, wherein the 5-FU-resistant colorectal cancer overexpresses transglutaminase 2.
Anticancer adjuvant for preventing or treating colon cancer, including streptonigrin and anticancer agents.
The anticancer adjuvant for preventing or treating colon cancer according to claim 8, wherein the anticancer agent is oxaliplatin or 5-FU.
The method of claim 8, wherein the Streptomyces you green and anticancer agent is from 1: anti-cancer adjuvant for cancer prevention or treatment, characterized in that contained as the molar concentration ratio of 10 ^5: 1 to 1.
Anticancer adjuvant for preventing or treating 5-FU-resistant colon cancer, including streptonigrin.
The anticancer adjuvant for preventing or treating 5-FU-resistant colon cancer according to claim 11, wherein the composition additionally contains 5-FU.
The anticancer adjuvant for preventing or treating 5-FU-resistant colon cancer according to claim 12, wherein the streptonigrin and 5-FU are contained in a molar ratio of 1: 1 to 1: 10 ⁶ .
The anticancer adjuvant for preventing or treating 5-FU-resistant colorectal cancer according to claim 11, wherein the 5-FU-resistant colorectal cancer overexpresses transglutaminase 2.

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