KR20200025221A - Composition for prevention, improvement or treatment of cancer comprising kalkitoxin as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating or treating cancer comprising kalkitoxin as an active ingredient, and specifically, to a pharmaceutical composition for preventing or treating the cancer, functional food composition for preventing or ameliorating the cancer, pharmaceutical composition for preventing or treating metastasis of cancer cells, and functional food composition for preventing or ameliorating metastasis of the cancer cells comprising kalkitoxin as an active ingredient. The composition can effectively prevent, ameliorate or treat the cancer by comprising kalkitoxin as an active ingredient. Particularly, the composition not only can effectively prevent, ameliorate or treat metastatic cancers generated as cancers are metastasized to regions other than initially generated regions, but also can effectively prevent, ameliorate or treat bone metastasis or metastatic brain tumor which is difficult to be treated compared to metastasis to general tissues among such metastatic cancers.

Description

Composition for prevention, improvement or treatment of cancer comprising kalkitoxin as an active ingredient}

The present invention relates to a composition for the prevention, improvement or treatment of cancer containing calkittoxin as an active ingredient, specifically, a pharmaceutical composition for the prevention or treatment of cancer containing calkittoxin as an active ingredient, functional for preventing or improving cancer Food compositions, pharmaceutical compositions for the prevention or treatment of cancer cell metastasis and functional food compositions for the prevention or improvement of cancer cell metastasis.

Despite many medical advances, cancer is still the leading cause of death. Drugs for preventing or treating cancer have been developed through various studies, but the development of new next-generation anticancer drugs is required as the development of carcinomas resistant to existing anticancer drugs and serious side effects of anticancer drugs are revealed.

Breast cancer, a type of cancer, is particularly famous for carcinomas with high metastasis. About 10% of breast cancer patients already have metastases at the time of diagnosis, and the median survival period does not exceed 24 months, despite the advancement and treatment effect of anticancer, resection and chemotherapy.

When cancer has spread to bones, it is accompanied by severe pain, such as pathological fractures or spinal nerve compression, which significantly reduces the quality of life. Currently, cancers that have metastasized to bone are being treated with bisphosphonate-based drugs or radiation and surgical procedures. However, bisphosphonate-based drugs are very difficult to administer and cause severe digestive system reactions, as well as pericarditis, headache, It causes side effects such as fever, endocrine system, gastrointestinal system and cancer.

Metastatic brain tumors are the most common form of brain tumors, and it is estimated that about 20 to 50% of patients with malignant tumors metastasize to the brain. Metastatic brain tumors use supplements such as steroids and mannitol to relieve symptoms, in addition to aggressive treatment with surgery or radiation. However, long-term use can cause many side effects with hormones. Existing chemotherapy for the treatment of brain metastatic cancer has the advantage of treating the whole brain lesion and cancer spread throughout the system at the same time, but the effect is very limited because there is a cerebrovascular barrier in the brain.

Nat Med. 2007 13 (1): 62-9. Cancer Sci. 2007 98 (1): 127-33. J Med Econ. 2013 16 (1): 19-29. Endocrinol Metab (Seoul). 2016 31 (4): 485-492. J. Proc Natl Acad Sci USA. 2005 27; 102 (39): 13909-14.

Therefore, the main object of the present invention is to provide a new composition that can effectively prevent, ameliorate or treat cancer, in particular metastatic cancer, in particular bone metastasis cancer or brain metastasis cancer.

Another object of the present invention to provide a composition that can effectively prevent, improve or treat the metastasis of cancer cells.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer containing calkitoxin as an active ingredient.

In the pharmaceutical composition for preventing or treating cancer of the present invention, the cancer is preferably metastatic cancer.

In the pharmaceutical composition for preventing or treating cancer of the present invention, the metastatic cancer is preferably bone metastasis cancer or brain metastasis cancer.

In the pharmaceutical composition for preventing or treating cancer of the present invention, the cancer is preferably breast cancer.

In the pharmaceutical composition for preventing or treating cancer of the present invention, the breast cancer is preferably triple negative breast cancer.

According to another aspect of the present invention, the present invention provides a functional food composition for the prevention or improvement of cancer containing calquitoxin as an active ingredient.

In the functional food composition for preventing or improving cancer of the present invention, the cancer is preferably metastatic cancer.

In the functional food composition for preventing or improving cancer of the present invention, the metastatic cancer is preferably bone metastasis cancer or brain metastasis cancer.

In the functional food composition for preventing or improving cancer of the present invention, the cancer is preferably breast cancer.

In the functional food composition for preventing or improving cancer of the present invention, the breast cancer is preferably triple negative breast cancer.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer cell metastasis containing calquitoxin as an active ingredient.

In the pharmaceutical composition for preventing or treating cancer cell metastasis of the present invention, the cancer cell metastasis is preferably bone metastasis or brain metastasis of cancer cells.

In the pharmaceutical composition for preventing or treating cancer cell metastasis of the present invention, the cancer cells are preferably breast cancer cells.

In the pharmaceutical composition for preventing or treating cancer cell metastasis of the present invention, the breast cancer cells are preferably triple negative breast cancer cells.

According to another aspect of the present invention, the present invention provides a functional food composition for the prevention or improvement of cancer cell metastasis containing calquitoxin as an active ingredient.

In the functional food composition for preventing or improving cancer cell metastasis of the present invention, the cancer cell metastasis is preferably bone metastasis or brain metastasis of cancer cells.

In the functional food composition for preventing or improving cancer cell metastasis of the present invention, the cancer cells are preferably breast cancer cells.

In the functional food composition for preventing or improving cancer cell metastasis of the present invention, the breast cancer cells are preferably triple negative breast cancer cells.

The composition of the present invention can effectively prevent, ameliorate or treat cancer by containing calkittoxin as an active ingredient. In particular, the composition of the present invention can effectively prevent, ameliorate or treat metastatic cancer caused by metastasis to a site other than the first site, as well as bone metastasis cancer and brain metastasis, which are difficult to treat compared to the general tissue. Mucosa can also be effectively prevented, ameliorated or treated.

Figure 1 shows the chemical structure of the calcitoxin, an active ingredient of the composition of the present invention.
Figure 2 shows the results of examining the effect of calcitoxin (KT) on cytotoxicity and cell growth rate. A, cytotoxicity test results; B, cell growth test results.
Figure 3 shows the effect of inhibiting cancer cell mobility according to the treatment concentration of calcitoxin (KT). A, wound healing assay results; B, results of a simple scratch test.
Figure 4 shows the effect of calcitoxin (KT) in the interaction between cancer cells and bone cells. A, the results of osteoclasts (osteoclast); B, experimental results on osteoblasts (osteoblast); NC, normal osteoclasts; OcCM, control group (osteoblast CM treatment group); OcCM_KT100, calquitoxin (100 nM) treated group (osteoblast CM and calkitoxin treated group).
Figure 5 shows the effect of calcitoxin (KT) on the expression of protein related to the mobility of cancer cells. A, Western blot experiments on the expression of HIF-1α, CaBP, Cathepsin K and MMP2 according to the concentration of calcitoxin treatment; B, Western blot experiments on the expression of p-38, p-JNK and p-ERK according to the calcitoxin treatment concentration.
Figure 6 shows a schedule for animal testing of calcitoxin (KT). Red arrow, period of taking kaquitoxin.
Figure 7 shows the effect of calkitoxin (KT) on cancer metastasized to bone. Left, IVIS image of experimental animals (top, 3 weeks; bottom, 5 weeks); Right, mean graph of femoral region of interest (ROI) values; PC Group, control group; KT Group, calkitoxin group.
Figure 8 shows the effect of calkitoxin (KT) on the recovery of bone damage caused by cancer. A, Micro CT image of femur of experimental animal; B, BMD and trabecular analysis graph after Micro CT scan; NC, normal group; PC, control group (cancer cell injection bone metastasis tumor model); KT1, calquitoxin 0.2 mg / kg administration group; KT2, calquitoxin 1 mg / kg administration group; KT3, calquitoxin 5 mg / kg administration group; PC + KT, calquitoxin 1 mg / kg dose group.
9 shows the effect of calkitoxin (KT) on cancers that have metastasized to the brain. Left, IVIS image of experimental animals (top, 3 weeks; bottom, 5 weeks); Right, mean graph of brain ROI (region of interest) values; PC Group, control group; KT Group, calkitoxin group.

The present invention has been devised based on new findings supporting that kalkitoxin can effectively prevent, improve or treat cancer, and metastasis of cancer cells can also be effectively prevented, improved or treated.

The chemical structure of chalcitoxin is shown in FIG. 1.

Animal testing using calkitoxin has shown that calquitoxin can effectively prevent, ameliorate or treat cancer.

In particular, animal studies can prevent, improve, or treat metastatic cancer, which is a carcinoma that has metastasized to other sites, by the administration of calquitoxin, and prevent, improve, or treat bone metastasis cancer and brain metastasis cancer, which is difficult to treat unlike general tissues It was confirmed that there is. In the case of brain metastasis, cerebrovascular barrier is difficult to deliver the drug, and accordingly the effect of chemotherapy is very limited. Calquitoxin has been confirmed that brain metastasis can be effectively prevented, improved or treated.

The composition of the present invention can exert an excellent effect on breast cancer, especially a carcinoma with a high metastasis rate, and has the advantage of preventing, improving, or treating triple negative breast cancer, which is very difficult to treat because no effective therapeutic agent has been developed so far. .

In vitro experiments with kalquitoxin confirmed that kalquitoxin could inhibit the mobility of cancer cells, and in particular, it was possible to effectively prevent, improve or treat cancer metastasis by inhibiting the interaction between cancer cells and bone cells. Confirmed.

The content of the calcitoxin as an active ingredient in the pharmaceutical composition of the present invention can be applied in various ranges according to the use or administration method, but will generally contain 0.1 to 90% by weight of the calcitoxin based on the total weight of the pharmaceutical composition of the present invention. I think you can.

The pharmaceutical composition of the present invention can be administered orally or parenterally during clinical administration and intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intrauterine dural injection, cerebrovascular injection or It can be administered by intrathoracic injection and can be used in the form of general pharmaceutical formulations.

It is contemplated that the pharmaceutical compositions 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 daily dosage of the pharmaceutical composition of the present invention may be about 0.1 to 10 mg / kg, preferably 0.5 to 2 mg / kg per day, based on the calcitoxin contained in the composition, and may be administered once to several times a day. However, the range will vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease.

In actual clinical administration, it may be administered in a variety of parenteral formulations, which may be formulated using conventional diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like may be used.

The pharmaceutical composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the calcitoxin.

The food composition of the present invention may be a composition mixed with calcitoxin itself or a food acceptable carrier based on the effective amount. The food composition of the present invention is processed meat products, fish products, tofu, jelly, porridge, noodles such as ramen or noodles, soy sauce, soybean paste, seasoned foods such as miso, red pepper paste, mixed soy sauce, dairy products, such as sweets, fermented milk or cheese, kimchi Foods such as pickles such as pickles, pickles, fruits, vegetables, soy milk, and fermented beverages may be possible. In addition, a pharmaceutically acceptable carrier may also be used as the pharmaceutically acceptable carrier.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by these examples.

<Example>

1. In vitro experiment

The purpose of this study was to determine the effect of calcitoxin on the invasiveness and mobility of cells by selecting concentrations that do not induce cytotoxicity.

1-1. Investigate effects on cytotoxicity and cell growth rate

As a result of exposure of calcitoxin (KT) to breast cancer cells (MDA-MB-231, triple negative breast cancer cell line) at concentrations of 25, 50, and 100 nM for up to 48 hours, the toxicity was confirmed. It is not shown. In addition, as a result of exposure to breast cancer cell lines for two weeks to confirm the effect of calkitoxin on the cell growth, as shown in B of FIG.

Accordingly, the verification of the efficacy of calkytoxin through in vitro experiments was performed according to the concentration of 100nM, exposure time within 48 hours.

1-2. Investigate breast cancer cell mobility inhibitory effect

In order to confirm the effect of calcitoxin (KT) on the mobility of tumor cells, a simple scratch test and a wound healing assay were performed. As a result, it was confirmed that the mobility of the cells statistically significant at 50nM and 100nM concentration as shown in FIG.

1-3. Investigation of the inhibitory effect of interaction between bone cells and tumor cells

In order for tumor cells to proliferate, it is very important to interact with tumor stem cells. Cancer cells that have metastasized to bone are further activated through mutual signaling with normal cells (osteoblasts or osteoclasts) present in the bone. The chemotaxis test between breast cancer cells and osteoblasts (or osteoclasts) was conducted to determine the effect of calcitoxin on the interaction between cancer cells and bone cells.

As a result of the chemotaxis experiment, as shown in FIG. 4, the cancer cells showed high chemotaxis to the substances secreted from osteoclasts (osteoclast, Oc) or osteoblasts (osteoblast, Ob). On the other hand, breast cancer cells exposed to CM (CM + KT) containing calkitoxin were found to have significantly reduced chemotaxis. These results suggest that calquitoxin may be involved in signaling between osteoclasts (or osteoblasts) and breast cancer cells.

1-4. Investigation of the expression regulation effect of tumor cell mobility-related protein

The effect of calcitoxin on the expression of proteins involved in the mobility of tumor cells was investigated by Western blot experiment.

As a result, the expression of CaBP, a protein known to inhibit cell mobility is increased in cells treated with calkittoxin (KT) as shown in FIG. 5, and known to increase cell mobility, such as Cathepsin K and HIF-1a. It was confirmed that the expression of the protein is reduced.

In addition, the major proteins related to MAPK, JNK, and ERK signaling pathways, which are well known for cell metabolism, were identified. As a result, JNK and ERK activities increased after 25 nM of calkitoxin treatment, and p38 decreased with concentration.

2. Animal Experiment

Bone metastatic tumor model was constructed using breast cancer cell line (MDA-MB-231, triple negative breast cancer cell line).

After confirming that the tumor was formed in the femur of the mouse, calcitoxin was orally administered for 2 weeks, and then the degree of tumor inhibition and bone recovery were verified by micro CT analysis and imaging technique.

2-1. Investigate the effects of metastatic cancer

After building a metastatic cancer animal model using the breast cancer cell line MDA-MB-231, the experiment was performed with the schedule as shown in FIG. Three weeks after the cancer cell line was injected into the mouse, the degree of metastasis to the bone or brain was checked, and calkitoxin (KT) was taken for two weeks to compare the change in cancer size and the extent of treatment of the damaged bone.

Cancer metastases were compared by the magnitude of the signal using an IVIS imaging system. Bone damage and recovery were verified by Micro-CT analysis.

As a result of IVIS imaging at 3 weeks (just before drug administration) and 5 weeks (2 weeks after drug administration), tumor signals in the thigh bones of the experimental group fed with calcitoxin (KT) were significantly higher than those of the control group as shown in FIG. 7. It confirmed that it decreased. This may be a result showing that the calcitoxin has a therapeutic effect on bone metastasis cancer.

As a result of comparative analysis of the degree of bone damage caused by metastatic cancer through the micro CT scan of the mouse thigh bone, as shown in FIG. 8, the bones damaged by cancer after the treatment with calcitoxin (KT) are treated as similar to those of normal mice. It showed an effect.

FIG. 9 is a graph analyzing IVIS photographs and signals of mouse brain sections at 3 weeks (just before drug administration) and 5 weeks (2 weeks after drug administration). In the control group, the tumor signal increased by 200-400 times between two weeks, whereas the experimental group fed with calkittoxin (KT) decreased to more than half the level.

Because of the brain's blood vessel barrier, the drug is difficult to pass, so chemotherapy has very limited effects. However, the results showed that the size of metastatic brain tumors was significantly reduced in the calquitoxin-treated group. This is significant as an indirect result showing that the metabolite of calquitoxin can cross the cerebrovascular barrier well.

Claims (10)

Pharmaceutical composition for the prevention or treatment of cancer containing calquitoxin as an active ingredient. The method of claim 1,
Pharmaceutical composition, characterized in that the cancer is metastatic cancer. The method of claim 2,
The metastatic cancer is a pharmaceutical composition, characterized in that the bone metastasis cancer or brain metastasis cancer. Functional food composition for the prevention or improvement of cancer containing calquitoxin as an active ingredient. The method of claim 4, wherein
Functional food composition, characterized in that the cancer is metastatic cancer. The method of claim 5,
The metastatic cancer is a functional food composition, characterized in that bone metastasis cancer or brain metastasis cancer. Pharmaceutical composition for the prevention or treatment of cancer cell metastasis containing calquitoxin as an active ingredient. The method of claim 7, wherein
The cancer cell metastasis is a pharmaceutical composition, characterized in that the bone metastasis or brain metastasis of cancer cells. Functional food composition for the prevention or improvement of cancer cell metastasis containing calquitoxin as an active ingredient. The method of claim 9,
The cancer cell metastasis is a functional food composition, characterized in that the bone metastasis or brain metastasis of cancer cells.

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