KR20030079484A - A Pharmaceutical Composition for Prevention or Treatment of a Disease associated with Inflammation Comprising Byakangelicin as an effective ingredient - Google Patents

Abstract

PURPOSE: Provided is a pharmaceutical composition for prevention or treatment of diseases associated with inflammation comprising byakangelicin as an effective ingredient. It is of an anti-inflammatory agent, and is thus useful for the prevention and treatment of septic shock. CONSTITUTION: A pharmaceutical composition for the prevention and treatment of diseases associated with inflammation is characterized by containing byakangelicin as an effective ingredient, wherein the byakangelicin is represented by the formula(1) and separated from the root of Ganoderma Sessile MURRILL., and the disease associated with inflammation is septic shock.

Description

A pharmaceutical composition for prevention or treatment of a disease associated with Inflammation Comprising Byakangelicin as an effective ingredient}

The present invention relates to an anti-inflammatory composition comprising villacan jellyline (byakangelicin), and more particularly, to a composition for preventing and treating inflammation-related diseases, in particular, for preventing and treating septic shock.

Inflammation is a phenomenon that occurs in damaged tissue caused by microbial infection, trauma, and the like, and causes many diseases. Several drugs have been used to control inflammation, which can be classified into steroidal anti-inflammatory drugs and nonsteroidal anti-inflammatory drugs. However, these drugs are often unsatisfactory in terms of effects and side effects in treating various inflammatory diseases, and thus, new drugs for suppressing the inflammatory response are continuously required.

Septic shock, initially accompanied by a severe systemic inflammatory response, results from an excessive systemic response of a host, such as a mammal, to bacterial infection, resulting in a mortality rate of about 45%. Specifically, this disease occurs because the host reacts excessively to endotoxins of Gram-negative bacteria. Conventionally, antimicrobial agents and steroidal agents have been used to treat septic shock, but the effect is negligible and the mortality rate of the host due to septic shock is still high.

The present inventors have conducted research for many years to develop an anti-inflammatory agent having no side effects and a high anti-inflammatory effect. As a result, it has been confirmed that villacanzelin is an anti-inflammatory agent far from its known therapeutic use. The present invention was completed by confirming the effects of treatment and prevention on shock.

It has never been reported that such villacanzelin has the effect of treatment and prevention against anti-inflammatory agents, in particular septic shock.

It is an object of the present invention to provide an anti-inflammatory agent containing villacan jellyline as an active ingredient.

Still another object of the present invention is to provide a composition for preventing or treating an inflammation-related disease including villacanjellyin.

Still another object of the present invention is to provide a composition for preventing or treating septic shock, including villacanjellisin.

1A to 1C are graphs showing variations in the levels of TNF-α, IL-6 and IL-10 over time in blood taken from experimental group 2 and control mice.

Figure 2 is a graph showing the GTP value measured in blood taken from experimental group 2 and control mice.

In accordance with the above object, the present invention provides a pharmaceutical composition comprising villacan jellyline as an active ingredient.

Hereinafter, the present invention will be described in detail.

Villacanzellicin is a pale yellow needle crystal having a molecular weight of 334 and is a compound having a structure of the following formula.

Such villacanjellisin can be isolated from the roots (plants) of plants, in particular, Angelica dahuria Benth, et Hook (Kiyoshi Hata, Mitsugi Kozawa, Kun-ying Yen (1963) Yakugaku Zasshi . 83, 606-). 610; Kuk Hyun Shin, Myung Sook Chung, Tae Soon Cho (1994) Arch. Pharm. Res. 17, 331-336).

Villacanlysin has been shown to prevent and treat inflammatory diseases. Such effects can be seen in experiments using animal models in which septic shock is artificially induced (see Examples).

Villacanlysin can be mixed with a suitable pharmaceutically acceptable carrier or excipient or diluted with a diluent according to conventional methods to prepare a pharmaceutical composition of the present invention. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The composition may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like.

The pharmaceutical compositions of the present invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.

The pharmaceutical compositions of the present invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular. Typical daily dosages of villacanjellisin as an active ingredient of the composition of the present invention range from 1 to 100 mg / kg, and can be administered once or in divided doses. However, it is to be understood that the actual dosage should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the patient, such dosages thus limiting the scope of the invention in any aspect. Not.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only for illustrating the present invention and are not intended to be limiting.

Preparation Example: Isolation of Villacanlizin from White Paper

9.6 kg of dried white paper was extracted three times with 10 L of methanol: water (7: 3) under reflux cooling at about 60 ° C. for 12 hours. The extract was filtered under reduced pressure and concentrated to obtain 3 kg of methanol extract. Distilled water (1.5 L) and n-hexane (1.5 L) were added to the methanol extract, and a fraction funnel was used to separate the n-hexane layer and the aqueous layer twice. The n-hexane layer was concentrated under reduced pressure to obtain an n-hexane extract. Then, the aqueous layer was extracted in the same manner as in the above ethyl acetate, butanol in order to obtain 710g n-hexane extract, 35.8g ethyl acetate extract, 128.95g n-butanol extract, 2.1kg water extract.

Among them, 35.8 g of ethyl acetate extract was subjected to silica gel column chromatography to separate compounds. That is, the column (length 80cm, diameter 8cm) was filled with silica gel (No.9385, 230-400mesh, Merck) about 21cm and eluted with 1L of n-hexane: ethyl acetate (6: 4) to saturate the stationary phase. After the extraction, 35.8 g of this extract was adsorbed onto about 45 g of silica gel (No.7734, 70-230mesh, Merck) and loaded on the column. flash starting with mobile phase elution with n-hexane: ethyl acetate (6: 4) to gradually elute the polarity of the ethyl acetate: methanol mixture while simultaneously flowing N ₂ gas through the column ) Fractions 1 to 16 were separated by column chromatography.

Fraction 11 of the separated fractions was dissolved in n-hexane: ethyl acetate and left at room temperature to obtain a substance, villacanjellyin, as a crystal.

The isolated and purified material was identified as Villacanzellicin based on the following physical and chemical properties and spectroscopic data (Kiyoshi Hata, Mitsugi Kozawa, Kun-ying Yen (1963) Yakugaku Zasshi . 83, 606-610). Kuk Hyun Shin, Myung Sook Chung, Tae Soon Cho (1994) Arch. Pharm. Res. 17, 331-336).

Reference Example: Low Volume LPS Mouse Model

The low-dose LPS mouse model is one of the septic shock animal models, in which lipopolysaccharide (LPS) is administered with D-galactosamine to significantly increase the mouse's sensitivity to LPS, resulting in hepatotoxicity even when low-dose LPS is administered. The septic shock of the back is caused.

The low-dose LPS model used in the present invention caused septic shock by administering 36 μg / kg LPS and 0.8 g / kg D-galactosamine into the abdominal cavity of ICR mice (20-30 g, 4-6 weeks old).

Example

Effects of Villacanzlysin on Low Dose LPC Models exam

Twenty ICR mice of the low dose LPC septic shock model according to the reference example were divided into four groups of five, and three groups of mice were isolated according to the preparation 30 minutes prior to administration of LPC / D-galactosamine. Villacanzelin was dissolved in 10% solution of DMSO and intraperitoneally administered at doses of 10 mg / kg, 30 mg / kg and 100 mg / kg (Experimental Group 1, Experimental Group 2 and Experimental Group 3), respectively, and the remaining 1 group of mice (control group). ) Was administered intraperitoneally with the same amount (0.1 ml) of a 10% solution of DMSO.

(1) survival rate measurement result

Survival rates of the experimental group 1, the experimental group 2, the experimental group 3 and the control mice over time were measured, and the results are shown in Table 1.

Table 1

24 hour survival rate 48-hour survival rate 72-hour survival rate Control 20 20 20 Experiment group 1 80 80 80 Experiment group 2 100 100 100 Experiment group 3 80 80 80

As can be seen in Table 1, in the experimental groups 1, 2, and 3 administered with villacanjellisin, mortality due to septic shock was lowered compared to the control group, and particularly, when villacanjellisin was administered at 30 mg / kg (experimental group) 2) All mice survived. Therefore, it was confirmed that Villacan Jellisin exerts a prophylactic and therapeutic effect of septic shock due to inflammation.

(2) Determination of the levels of cytokines TNF-α, IL-6 and IL-10

When inflammation occurs, the levels of the inflammatory cytokines TNF-α, interleukin-6 (IL-6) and the anti-inflammatory cytokine interleukin-10 (IL-10) increase in the blood. Therefore, measuring the levels of these cytokines in the blood can confirm the inhibitory effect of inflammation.

After intraperitoneal administration of LPC / D-galactosamine from ICR mice of Experimental Group 2 and Control Group, blood was collected at 90 minutes, 3 hours, and 6 hours, and serum was separated using a centrifuge, followed by ELISA kit. The levels of TNF-α, IL-6 and IL-10 in the serum were measured, and the results are shown in FIGS. 1A to 1C.

As can be seen in Figure 1a, the blood of the experimental group 2 mice significantly reduced TNF-α, an inflammatory cytokine. In other words, TNF-α, a major causative agent of sepsis (inflammation), was drastically reduced when villacanjellisin was administered, and mortality due to septic shock was excellently reduced. In addition, there was no significant change in the levels of IL-6 and IL-10 in the blood of Experimental Group 2 (FIGS. 1B and 1C).

(3) hepatotoxicity test

LPC / D-galactosamine was injected intraperitoneally from mice of the experimental group 2 and the control group, and 0.5 ml of blood was collected from the bile vein at 90 minutes, and serum was separated by centrifugal separation. The level of alanine aminotransferase (ALT), an indicator, was measured according to the Reitman-Frankel method (Reitman S. and Frankel S., Am. J. Clin. Pathol . 28: 56-63, 1957). The results are shown in FIG.

As can be seen in FIG. 2, the ALT value, an indicator of hepatotoxicity, was significantly reduced in the experimental group 2 mice compared to the control group.

According to the present invention, the pharmaceutical composition containing villacanjellyin has an anti-inflammatory effect, and thus can be usefully used for preventing and treating inflammation-related diseases, especially septic shock.

Claims (4)

Anti-inflammatory agent containing villacan jellyline as an active ingredient. Pharmaceutical composition for the prevention and treatment of inflammation-related diseases containing villacan jellyline as an active ingredient. The pharmaceutical composition according to claim 2, wherein the inflammation-related disease is septic shock. The anti-inflammatory or pharmaceutical composition according to any one of claims 1 to 3, wherein the villacan jellyline is isolated from the root of white paper.