KR101288894B1 - Composition for the prevention and treatment of cancers containing CKD712, (S)-1-(α-naphthylmethyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline or its pharmaceutically acceptable salts - Google Patents

Abstract

The present invention provides CKD712 (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof as an active ingredient It relates to an anticancer pharmaceutical composition comprising and an anticancer activity enhancer. The compounds inhibit the growth and metastasis of cancer cells by regulating PTEN and Akt related signaling systems.

Description

COD7112, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof as an active ingredient Composition for the prevention and treatment of cancers containing CKD712, (S) -1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline or its pharmaceutically acceptable salts }

The present invention provides CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof as an active ingredient. It relates to a pharmaceutical composition for anticancer and anticancer activity enhancing agent comprising a.

Cancer (tumor) is the product of abnormal, uncontrolled and disordered cell proliferation. If there is destructive growth, invasiveness and metastasis of such tumors, they are classified as malignant. Invasiveness is the property of invading or destroying surrounding tissues. Invasiveness generally means that the tumor spreads locally by destroying the basal layer that forms the boundary of the tissues, and often enters the circulatory system of the body. Tumor usually refers to the spread of tumor cells by lymphatic or blood vessels to a different location from the primary site. It also means to move tumor cells by elongation directly through serous body cavity or other spaces.

Currently, cancer is mainly treated through one of three treatments: surgical surgery, radiation and chemotherapy, or a combination thereof. Surgery involves removing most of the diseased tissue. Such surgical operations are effective in removing tumors located in certain areas, such as breasts, colon and skin, but cannot be used to treat tumors in some areas, such as the spine, or to treat diffuse tumor diseases such as leukemia. Chemotherapy disrupts cell replication or cell metabolism and is often used to treat cancers of the breast, lungs, and testes. Side effects of systemic chemotherapy used to treat tumor diseases are most problematic for patients receiving cancer treatment do. Among these side effects, motion sickness and vomiting are the most common and serious side effects. Side effects from chemotherapy have a profound effect on the patient's life and can dramatically change the patient's compliance with treatment. In addition, side effects associated with chemotherapeutic agents generally include a major dose limiting toxicity (DLT) that should be exercised when administering such drugs. For example, mucositis is major dose limiting toxicity to several anticancer agents, such as 5-Fluorouracil, methotrexate, and anti-tumor antibiotics (eg doxorubicin), which are anti-metabolite cell toxins. Most of these side effects from chemotherapy require hospitalization in severe cases or require analgesics to treat pain. As such, side effects caused by chemotherapeutic agents and radiation therapy are becoming a major problem in the clinical treatment of cancer patients. In addition, conventional anticancer drugs often kill up to normal cells, and thus, researches on new mechanisms of anticancer drugs that can act only on cancer cells, not on normal cells, have recently been conducted.

Breast cancer is the most common cancer in the United States, which ranks first among female cancers and third in Korea. About 20,000 women get breast cancer each year, and the number of patients increases every year. Breast cancer can be treated by surgical resection for early diagnosis, but many patients relapse in the form of distant metastasis such as lung, bone, and liver. More specifically, in Korea, the incidence of breast cancer was only 8.7% of female cancers in 1984. However, in 2000, 15.1% of female cancers appeared, followed by 15.6% of stomach cancers. Not only does it increase numerically, but the rate of increase is even more concerned. Its incidence has increased by more than 50% in just 15 years, and breast cancer is expected to be the most common cancer among women in the near future. In addition, Asians generally have a lower frequency of breast cancer than Westerners. According to the statistics in 2000, compared with 23/10/10 people of Korean breast cancer and 20-30 people / 100,000 Japanese, 111.8 / 100,000 Americans (whites) and 95.4 / 100,000 Americans (blacks) . While Westerners are more likely to develop breast cancer as they get older, breast cancer in Korea is most common in their 40s, followed by those in their 50s, 60s and 30s.

There are many possible causes of breast cancer, but the most important is the female hormone, but also has a large genetic impact. In the case of breast cancer in the family, genetic testing can confirm that the incidence of some gene variants is high. In addition, fatty foods are recognized as another cause.

On the other hand, paclitaxel (paclitaxel or taxol) is extracted from the bark of Taxus brevifolia native to the west coast of the United States, and is considered to be one of the most important drugs in chemotherapy of cancer. Paclitaxel selectively acts on tubulin without affecting the synthesis of DNA and RNA in cancer cells and without damaging the DNA molecule itself. This stops the growth of the polymerized tubulin by depolymerization and stops the growth of the cancer cells during the cell division, thereby killing the cancer cells. Although paclitaxel has recently been used as a chemotherapeutic agent for breast cancer, it has not been used for the treatment of advanced cancer due to drug resistance.

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline), has been studied as a potential anti-immune agent. Past studies have shown that CKD712 can be used in the treatment of immune disorders such as sepsis, and has been shown to inhibit atherosclerosis by inhibiting adhesion molecules caused by inflammatory cytokines. However, it is not known until now that CKD712 has anticancer activity.

Accordingly, the present inventors have tried to develop a novel anticancer agent, and as a result, CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) The present invention was completed by enhancing the anticancer activity of paclitaxel, which is achieved through the PTEN / PI3K / Akt signaling system.

Domestic application number: 10-2009-0074870

The present invention provides an active ingredient of CCD71, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof. To provide a pharmaceutical composition for cancer comprising.

The present invention also relates to CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof. To provide an anticancer activity enhancer comprising as an active ingredient.

In order to achieve the above object, the present invention provides CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically thereof. It provides a pharmaceutical composition for anticancer comprising an acceptable salt as an active ingredient.

The present invention is also effective for CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof. It provides an anticancer activity enhancer comprising as a component.

The compounds of the present invention inhibit the growth and metastasis of cancer cells by regulating PTEN and Akt related signaling systems, and play a role in enhancing the activity of anticancer agents. Therefore, the use of the composition of the present invention can increase the therapeutic effect while reducing the anticancer toxicity by using a small amount of anticancer agents.

1 is a graph confirming the effects of CKD712 and paclitaxel on cell survival.
2 shows the results of MTT assay with increasing concentration of CKD712.
3 and 4 are pictures showing the effect of paclitaxel and CKD712 on cell metastasis.
5 and 6 illustrate the effects of CKD712 on Akt and PTEN protein activity.
7 is a diagram confirming the relationship between the inhibitory effect and PTEN on the survival and metastasis of paclitaxel cells.
8 is a graph confirming the effects of paclitaxel and PTEN on cell survival.
9 and 10 are graphs confirming the effects of paclitaxel and PTEN on cell metastasis.
11 is a diagram confirming the effect of LY294002 (PI3K inhibitor) on the expression of MMP9.
12 is a graph confirming the effect of LY294002 (PI3K inhibitor) on the cell survival inhibitory effect of paclitaxel.
13 and 14 are graphs confirming the effect of LY294002 (PI3K inhibitor) on the cell metastasis inhibitory effect of paclitaxel.
15 is a diagram confirming whether the effect of CKD712 on the sensitivity of paclitaxel through the PTEN activity, comparing the degree of expression of proteins (MMP9, Akt, PTEN).
Figure 16 shows the effect of CKD712 on the sensitivity of paclitaxel via the PTEN activity using siRNA, confirmed by MTT assay.

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

The present invention provides CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof as an active ingredient. It provides a pharmaceutical composition for cancer comprising.

The active ingredient of the present invention is CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline), the tetrahydroisoquinoline compound Pharmaceutically acceptable salts thereof or prodrugs thereof.

The salt of the tetrahydroisoquinoline compound means a conventional salt used pharmaceutically, and various acids used in the preparation of the salt include hydrochloric acid, bromic acid, sulfuric acid, methanesulfonic acid, propionic acid, succinic acid, glutaric acid, citric acid, Fumaric acid, maleic acid, tartaric acid, glutamic acid, gluconic acid, glucuronic acid, galacturonic acid, ascorbic acid, carbonic acid, phosphoric acid, nitric acid, acetic acid, L-aspartic acid, lactic acid, vanic acid and hydroiodic acid.

As the functional derivatives of the prodrug compounds of the present invention, they can be easily changed to enter the living body and to show efficacy. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described in the existing literature.

The pharmaceutical compositions of the present invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral dosage forms, sterile injectable solutions, suppositories, and transdermal formulations according to conventional methods. have. Carriers, excipients and diluents that may be included in the composition 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. If necessary, it is formulated with diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like.

In one embodiment, the tetrahydroisoquinoline compounds of the present invention may be formulated as solid preparations for oral administration. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which include at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like. It is formulated by mixing. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

In another embodiment, the pharmaceutical composition containing the tetrahydroisoquinoline compound of the present invention may be formulated into a liquid preparation for oral administration. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, and the like. In addition to conventional inert diluents (e.g., purified water, ethanol, liquid paraffin), various excipients may be used. Wetting agents, sweetening agents, fragrances, preservatives and the like can be included.

In another embodiment, the pharmaceutical composition containing the tetrahydroisoquinoline compound of the present invention may be formulated into a formulation for parenteral, preferably intraperitoneal administration. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the sterilized aqueous solution, a suitable buffer solution such as Hank's solution, Ringer's solution or physically buffered saline can be used. As the non-aqueous solvent, propylene glycol, polyethylene glycol, olive oil, The same vegetable oil, an injectable ester such as ethyl oleate, or the like can be used. If desired, preservatives, stabilizers, wetting or emulsifying agents, salts for controlling osmotic pressure and / or buffers may be used. Meanwhile, in the case of suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, and the like, which are conventional bases thereof, may be used.

Compositions formulated in such a manner can be administered in various amounts, including orally or parenterally (percutaneously, subcutaneously, intravenously, intramuscularly, intraperitoneally) in an effective amount. As used herein, an "effective amount" refers to an amount exhibiting a prophylactic or therapeutic effect when administered to a patient. The dosage of the composition according to the present invention may be appropriately selected depending on the route of administration, subject of administration, age, sex, weight, individual difference and disease state. Preferably, the composition of the present invention may vary the content of the active ingredient according to the degree of disease, preferably 1 μg ~ 100mg / weight kg / day, more preferably 10 μg ~ 10mg / weight kg / day It may be administered in an effective amount of.

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

The present invention is also effective for CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) or a pharmaceutically acceptable salt thereof. Provided is an activity enhancer of an anticancer agent comprising as a component.

Subject cancer to which the composition of the present invention can be applied are lung cancer, pancreatic cancer, non-small cell lung cancer, liver cancer, colon cancer, bone cancer, skin cancer, head cancer, cervical cancer, skin or intraocular melanoma, cervical cancer, uterine cancer, endometrial carcinoma, vagina Carcinoma, vulvar carcinoma, fallopian tube carcinoma, ovarian cancer, rectal cancer, anal muscle cancer, gastric cancer, breast cancer, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis cancer, prostate cancer, bladder cancer , Renal cancer, ureter cancer, renal cell carcinoma and renal pelvic carcinoma and the like, preferably breast cancer.

The anticancer agent that can be used with the composition of the present invention is not particularly limited as long as it is a drug that inhibits the proliferation or metastasis of cancer, and anthracycline anticancer antibiotics (doxorubicin, daunorubicin, epirubicin, darvisinvinca alkaloid, etc.) ), Taxane-based anticancer agents (paclitaxel, docetaxel, etc.), colchicine, epipodophyllotoxin-based anticancer agents (etoposide, teniposidein, etc.), mitosantron, actinomycin D, and mitomycin C, and the like. Preferably paclitaxel.

Hereinafter, preferred embodiments and experimental examples are provided to facilitate understanding of the present invention. However, the following examples and experimental examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples and experimental examples.

Example  1: anti-proliferation ( anti - proliferative Effect Enhancement Experiment

MTT assay was performed to determine the effect of CKD712 on the survival of human breast cancer cells (MDA-MB-231).

As a result, as shown in Figure 1, paclitaxel showed a synergistic effect of inhibiting significant cell growth concentration-dependently when treated with CKD712. In addition, as shown in FIG. 2, cell growth was confirmed when CKD712 was treated alone or after treatment with Paclitaxel. As a result, CKD712 showed a significant inhibitory effect on cell growth and increased the inhibitory effect of paclitaxel. However, when CKD712 was treated at low concentration, this enhancement effect was not significant.

Example  2: anti-transition ( anti - metastatic Effect Enhancement Experiment

Invasion assay was performed using MDA-MB-231 cells.

As a result of the experiment, as shown in Figs. 3 and 4, the movement of the cells treated with Paclitaxel was weakened, whereas the cells without any treatment (con) passed through the metrigel. In addition, when CKD712 and Paclitaxel were treated together, it was confirmed that the anti-migration effect of Paclitaxel was increased. CKD712 alone also inhibited cell migration compared to the control.

In addition, previous studies have shown that PI3K / Akt signaling plays an important role in cell growth and invasion, confirming whether CKD712 is involved.

5 and 6, it was confirmed that CKD712 inhibits its activity by dephosphorylation of Akt and regulates the phosphorylation of PTEN to affect PTEN activity. CKD712 also regulates matrix metalloproteinase-9 (MMP9), a well-known pro-metastatic protein. In particular, treatment with Paclitaxel alone did not affect p-Akt, p-PTEN, and MMP9dml activities, and significant results were obtained only when combined with CKD712.

Example  3: PTEN Of PI3K Of Akt  Analysis of the impact on signal transmission

In Example 2 it was confirmed that CKD712 activates PTEN in MDA-MB231 breast cancer cells. Therefore, this study was performed using PTEN overexpression vector to explain the importance of PTEN on the anticancer effect of Paclitaxel in breast cancer cells.

As shown in FIG. 7 and FIG. 8, consistent with the results of the previous study, overexpression of PTEN significantly reduced the expression of MMP9 protein and resulted in inhibition of its active cell migration. In addition, as shown in FIGS. 9 and 10, the effect of Paclitaxel (activity of PTEN, p-Akt, MMP9 and cell migration) was significantly increased in cells transformed with PTEN overexpression vector.

Example  4: PI3K Of Akt  Analysis of the impact on signal transmission

In order to demonstrate that the effect of CKD712 to enhance the efficacy of Paclitaxel in breast cancer cells was derived from the activity of PTEN / PI3K / Akt, LY294002 was treated to inhibit PI3K / Akt signaling.

9 to 13, LY294002 was confirmed to enhance the action of Paclitexel by inhibiting cell growth, matrigel invasion and MMP9 protein expression in breast cancer cells.

Example  5: PTEN  Activity analysis

Experiments were performed using SiRNA to reaffirm that CKD712 regulates the activity of PTEN.

As shown in Figure 14 to 16, when Paclitaxel alone treatment did not affect the expression of PTEN, p-Akt, MMP9. However, when cells were transformed with siRNA, CKD712 markedly reduced the expression of p-Akt and MMP9 regardless of the presence of Paclitaxel. However, the cells transformed with siPTEN did not show any sensitivity to the treatment of CKD712, indicating that CKD712 mediates the growth, infiltration, and inhibition of MMP9 expression regulation through PTEN.

Hereinafter will be described a pharmaceutical composition containing the composition of the present invention and a preparation example of a health functional food, but is not intended to limit the present invention only intended to be described in detail.

Formulation example  One. Sanje  Produce

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3. Preparation of capsules

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

Formulation example  5. Liquid  Produce

CKD712, (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Claims (6)

delete delete An activity enhancer of paclitaxel, which is an anticancer agent comprising CKD712 (S-1- (α-naphthylmethyl) -6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) as an active ingredient. delete delete 4. The activity enhancer of paclitaxel, which is an anticancer agent, according to claim 3, wherein the activity enhancer has an activity of inhibiting the growth and metastasis of breast cancer cells.

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