KR980008225A - Anticancer drugs containing dexacinol angelate as an active ingredient - Google Patents

Abstract

The present invention relates to an anticancer agent comprising decursinol angelate as an active ingredient, and it has been newly confirmed that decercinol angelate has an anticancer activity against various cancer cell lines such as lung cancer and liver cancer cells, To a method for producing an anticancer agent.

Description

Anticancer drugs containing dexacinol angelate as an active ingredient

Figure 1 shows the results of measuring the bio-induction of decercanol angelate against human lung cancer cell line (A-549), chronic white blood cell line (K-562), liver cancer cell line (Hep G ₂ ) and normal lung cell line (MRC-5) Fig.

[TECHNICAL FIELD OF THE INVENTION AND RELATED ART OF THE SAME]

The present invention relates to decursinol angelate, which is effective as an anticancer agent, and more specifically, decursinol angelate represented by the following formula (1) is newly found to have anticancer activity against various cancer cell lines. To an anticancer agent comprising the active ingredient.

The decursinol angelate represented by the above formula (1) can be prepared by the method of the present invention using a pyranocoumarin-based natural material component (Khim. Prir. Soedin (R)) first isolated from the extract of Seseli grandivitattum in Russia 5, 640 (1977)), but its pharmacological activity has yet to be known.

In addition, decercinol angelate was extracted from Korean Angelica gigas Nakai (Korean J. Pharmacogn., 21 , 64-68 (1990)), but decercinol angelate was extracted from other plants Is not known.

The pharmacokinetics and biological properties of tachycinol angelate extracted from Korean angelica orientalis (Angelica gigas) are unknown at present. It is a structural isomer of dexacinol angelate and decursine extracted from Korean angelica orientalis, like dexacinol angelate, paralyzes the ejected intestines of the rabbit and the extracted heart of the frog, , And it is known to act relatively excitingly on the rabbit's removed uterus (see Herbal Medicine Vol. 1, No. 1, pp. 25-32, 1970).

[Technical Problem]

The present inventors have long been studying to construct a biological or biochemical system capable of easily detecting an anticancer agent from a plant or a herbal medicine, and have found that decercinol angelate represented by the above formula (1) Toxicity, and formulated it as an anticancer agent, thereby completing the present invention.

[Structure and operation of the invention]

The present invention is characterized by an anticancer agent comprising decacinol angelate represented by the above formula (1) as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention relates to the use of a new deacercinol angelate used for the treatment of hepatocellular carcinoma, lung cancer, chronic leukemia and the like by containing decercinol angelate as an active ingredient.

The decancinol angelate represented by the above formula (1) used in the present invention is a compound or a natural product extracted from Korean Angelica gigas Nakai.

First, the method for preparing decancinol angelate from Korean Angelica angustifolia is prepared by using ethanol as an extraction agent as a conventional method (Korean J. Pharmacogn., 21 , 64-68 (1990)).

In the present invention, a drug is prepared using decancinol angelate prepared as described above as an active ingredient. In the present invention, the pharmaceutical composition may be either oral or parenteral.

The formulations may be in a form suitable for administration via various routes including oral, rectal, transdermal, subcutaneous, intravenous and intramuscular, for example, tablets, hard and soft gelatine capsules, solutions, emulsions, suspensions, And the like.

For the preparation of pharmaceutical preparations, the compounds may be combined with therapeutically inert inorganic or organic carriers.

If it is formulated into tablets or economic gelatin capsules in oral form, lactose, corn starch or derivatives thereof, active stearic acid or its salts may be used as carriers.

Examples of suitable carriers for soft gelatine capsules are vegetable oils, waxes, fats, semi-solid and liquid polyols and the like, although carriers may not be required in some cases.

In addition, examples of carriers suitable for the preparation of liquids and syrups include water, alcohols, polyols, glycerin or vegetable oils.

In the case of parenteral injection preparation, D-mannitol, purified water, alkali metal salt or p -hydroxybenzoate may be used as the carrier.

Other suitable carriers for the manufacture of suppositories are natural and hardened oils, waxes, fats, semi-liquid polyols and the like.

On the other hand, the pharmaceutical preparations may contain preservatives, stabilizers, wetting agents, emulsifiers, sweeteners, coloring agents, flavors, salts for controlling osmotic pressure, buffers, coating agents or antioxidants.

When used as a cancer treatment, the dosage can vary widely and, of course, is adjusted according to individual requirements in each particular case. Generally, when administered to an adult, a daily dosage of about 1 to 50 mg, preferably about 7 mg per kilogram of body weight is adequate, but in some cases it may exceed the upper limit. The daily dose may be administered in a single dose or divided doses.

Hereinafter, the effect of decancinol angelate extracted from Korean angelica orientalis Angelica angelica and the method of preparing the anticancer activity will be described in more detail with reference to the following examples. As a result, the effect is particularly excellent compared with decancinol angelate used in the present invention The compounds of the present invention are not limited to those described in the Examples but are not limited thereto.

[Example 1]

Isolation of Dacercinol Angelate from Angelica giganteus Ethanol Extract

Korean ginseng Angelica edulis (400 g) was placed in a flask, shaken at 50 ° C for 2 days with 1 L of ethanol, and filtered to remove insoluble matter. This ethanol extract was used to separate the dexacinol angelate of the coumarin family through the following multi-stage separation process.

1 L of the ethanol extract obtained above, from which the insoluble matter was removed, was distilled under reduced pressure and concentrated. To remove sugars or other water-soluble substances, 500 ml of hexane and distilled water were added to the concentrate, and the mixture was stirred. After separating each layer, the water layer was discarded and the hexane layer was further distilled under reduced pressure. Subsequently, ethyl ether and toluene were mixed at a ratio of 1: 4, 30 ml was taken, the concentrate was suspended, and silica gel chromatography was repeatedly carried out using the same solvent as that of the suspension. The fractions obtained through silica gel chromatography were combined and distilled under reduced pressure. 3 g of blue fluorescent substance was obtained under ultraviolet light through this process. The analysis was carried out using high performance liquid chromatography (HPLC) And deckerin. The deacercinol angelate was separated using a normal phase high performance liquid chromatography using a preparative column for pure separation.

Infrared spectroscopy, nuclear magnetic resonance analysis ( ¹ H-NMR, ¹³ C-NMR) and mass spectrometry were performed on the obtained material. The results are as follows.

_{IR (KBr, λ max) cm} -1: 2960 (CH), 1725 (C = O), 1630, 1653, 1495 ( aromatic ring), 1390, 1380 ((CH 3) 2 C =) 1 H-NMR ( _{300 MHz, CDCl 3) δ ppm} : 1.38 (3H, s, Me-2 '), 1.40 (3H, s, Me-2'), 1.89 (3H, d, J = 7.2 Hz, Me-4 '') , 1.84 (3H, d, J = 1.4 Hz, Me-2 ''), 2.90 (2H, dd, J = 17.6, 4.8 Hz, H-4 '), 3.23 (2H, dd, J = 17.6, 4.8 Hz (1H, d, J = 9.5, H-3 '), 5.13 (1H, t, J = 4.8, H- , 6.80 (1H, s, H -5), 7.16 (1H, s, H-8), 7.59 (1H, d, J = 9.5, H-4), 13 C-NMR (75.5 MHz, CDCl 3) δ ppm: 20.5 (2 " -Me), 23.2 (2 ' -Me), 25.1 (2 ' -Me), 15.8 ), 77.1 (C-3 '), 104.6 (C-8), 112.8 (C-10), 113.3 5), 143.1 (C-4), 154.2 (C-9), 156.4 (C-7), 139.5 MS m / e: 328 (M ⁺ ), 228 (xanthyletine), 213 (benzopyrylium), 83 (angeloyl acid)

As a result of the above-mentioned analysis results, it was confirmed that the extract had decancinol angelate of molecular formula C ₁₉ H ₂₀ O ₅ .

[Example 2]

Anticancer effect of decercinol angelate on human lung cancer cell line (A-539, ATCC CCL 185)

Decercinol angelate isolated in accordance with Example 1 was dissolved in dimethylsulfoxide (DMSO) to a concentration of 10 mg / ml prior to addition to the cell culture medium.

The A-549 cell line was cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% combined antibiotic (penicillin, streptomycin), and decancinol angelate Subculture was performed for about one week or more before treatment. When decancinol angelate was treated, the concentration of A-549 cells was adjusted to 2 × 10 ⁵ cells / ml in a 25 cm 2 -tissue flask, and a total of 10 ml was made. The amount of dimethyl sulfoxide added at the time of treatment was constant in all experimental groups including the control, so that the total amount of dimethyl sulfoxide was not more than 0.5%.

After treatment with decussinol angelate at 0 to 10 μg / ml, the cells were incubated at 37 ° C. in a 5% carbon dioxide incubator for 2 days, and the number of viable cells in each experimental group was measured under an inverted microscope. As a result, 1, and stained with Trypan blue to confirm viability.

From FIG. 1, it was found that the concentration of decascinol angelate, that is, the ED ₅₀ (50% cell lethal concentration), which showed a survival rate of 50% as compared with the control without treatment with decascinol angelate, / Ml, indicating significant A-549 cell lethal effects by deacercin angelate.

[Example 3]

Anticancer Effect of Deckerinol Angelate on Human Chronic Leukemia Cell Line (K-562, ATCC CCL 243) In the same manner as in Example 1, the anti-cancer effect of decercinol angelate on the chronic leukemia cell line K-562 The effect was measured by adding 10% fetal bovine serum and 1% complex antibiotic (penicillin, streptomycin) to RPMI-1640 medium.

As shown in FIG. 1, when the concentration of decancinol angelate was in the range of 0 to 20 μg / ml, the ED ₅₀ value of decancinol angelate for the K-562 cell line was found to be 15 μg / , Decercinol angelate was also found to have a strong anti-cancer activity in the K-562 cell line.

[Example 4]

Anticancer effect of decaccinol angelate on human hepatoma cell line (Hep G ₂ , ATCC HB 8065)

Even drugs that have efficacy in the in vitro (in vitro) test lot if the living body (in vivo) test efficacy does not appear, this is known to be due to detoxification and immune system by the liver largely. Thus, the present inventors have found that the efficacy of the decker resorcinol Ansel rate for Hep G ₂ cell line derived from the experiment were carefully cross. The culture medium was prepared by adding 10% fetal bovine serum and 1% mixed antibiotic (penicillin, streptomycin) to DMEM (Dulbecco's Modification of Eagle's MEM).

1, the ED ₅₀ value of decancinol angelate for the Hep G ₂ cell line was found to be 10 μg / ml , Indicating that decursinol angelate is not inactivated by hepatic function, and that anticancer activity against Hep G ₂ cell line is also excellent.

[Example 5]

Effect of decascinol angelate on human normal lung cells (MRC-5, ATCC CCL 171)

As an experiment to compare the effect of decascinol angelate with the cancer cell lines tested in the above example, the effect of decascinol angelate on human normal cell, MRC-5, was measured.

In order to measure the effect of decancinol angelate on MRC-5 cells, the same experiment as in Example 1 was carried out. In this case, DMEM (Dulbecco's Modification of Eagle's MEM) was supplemented with 10% fetal bovine serum and 1% A medium supplemented with antibiotics (penicillin, streptomycin) was used.

As shown in FIG. 1, the ED ₅₀ value of decancinol angelate for MRC-5 cells was measured at about 50 μg / ml, and it was confirmed that it exhibited a much lower lethal effect than the lethal effect for the cancer cell lines .

Therefore, it can be seen that dexacinol angelate can be used as a preferable anticancer drug in view of the fact that it acts specifically on cancer cells and has low toxicity to normal cells as an anticancer drug.

[Example 6]

Manufacture of tablets

5 g of decancinol angelate was homogeneously mixed with 10 g of lactose and 0.1 g of magnesium stearate, and 500 mg of these were partially molded into tablets.

[Example 7]

Preparation of capsules

A formulation in the form of a gelatin hard capsule was prepared containing 100 mg of decascinol angelate per capsule, 100 mg of lactose, 30 mg of starch, 2 mg of magnesium stearate and 17 mg of talc.

[Example 8]

Manufacture of liquid agent

30 g of decacyinol angelate and 50 g of D-mannitol were dissolved in about 600 ml of distilled water with stirring. 0.5 g of p - hydroxybenzoate and 2 g of alkali metal salt were dissolved in the resulting solution, and distilled water was added to prepare 1,000 ml of the solution. The solution was filtered and filled with ampoules in an amount of 4 ml per each ampoule according to a conventional method.

[Effects of the Invention]

As described above, the anticancer activity of decancinol angelate is disclosed in the present invention, and it can be formulated into an active ingredient to provide drugs useful for the treatment of lung cancer, liver cancer, and chronic leukemia.

Claims (5)

An anticancer agent comprising decaccinol angelate represented by the following formula (1) as an active ingredient. 2. The anticancer agent according to claim 1, wherein the deacercinol angelate is selected from the group consisting of a compound and a natural product extracted from Korean Angelica gigas Nakai. The anticancer agent according to claim 1, wherein the anticancer agent is a therapeutic agent for liver cancer. The anticancer agent according to claim 1, wherein the anticancer agent is a therapeutic agent for lung cancer. The method of claim 1, wherein the anticancer agent is a therapeutic agent for chronic leukemia. ※ Note: It is disclosed by the contents of the first application.