KR20090022928A - Composition for curing sepsis comprising compound k - Google Patents

Abstract

A pharmaceutical composition comprising Compound K(transformant of a red ginseng saponin) is provided to prevent and treat septic diseases caused by Gram negative bacteria using the Compound K obtained by decomposing Ginsenoside Rb1, Rb2, Rc and Rd with a cellulase enzyme. A pharmaceutical composition for treating septic diseases comprises a Compound K represented by the formula 1. The Compound K is a transformant of Ginsenoside Rb1, Rb2, Rc and Rd, which are saponin of red ginseng. The Compound K is prepared by decomposing the Ginsenoside in the presence of Aspergillus niger-derived cellulose. The septic diseases are caused by LPS(Lipopolysaccharide) of Gram negative bacteria.

Description

Composition for treating sepsis containing compound VII {Composition for curing sepsis comprising Compound K}

The present invention relates to a composition for treating sepsis containing Compound K, and more particularly, to a composition for treating sepsis containing Compound K (Compound K) represented by Formula 1 as an active ingredient.

Gram-negative sepsis is one of the leading causes of death in the intensive care unit of hospitals, and its frequency continues to increase worldwide due to increased frequency of infiltration pathways and treatments that lead to immunosuppression. . This symptom is characterized by damage to endothelial cells, coagulation disorders, vasospasm, tissue hypoperfusion, and impaired function of various organs. This is the result of a massive, uncontrolled inflammatory response due to the substances produced by the microorganisms. These substances include endotoxins called lipopolysaccharides (LPS), which are components of the outer membrane of Gram-negative bacteria and cause the production of proinflammatory cytokines. Except for the administration of antibiotics, treatment for sepsis is very limited. Therefore, there is a great demand for finding a substance that improves the survival rate of sepsis hosts and suppresses systemic inflammation.

Ginseng has long been used for health. Ingredients that indicate the pharmacological efficacy of ginseng are ginsenosides (ginsenosides), which exhibit various biological effects, such as anti-inflammatory and anti-tumor. Among these ginsenosides, ginsenosides Rb1, Rb2, Rc and Rd are metabolized by enterobacteria and converted to compound K (compound K, C-K; FIG. 1).

Compound K (20-O-beta-D-glucopyranosyl-20 (S) -protopanaxadiol) is a metabolite of protopanaxadiol ginsenoside, which acts as a variety of immunological pharmacological agents (Wakabayashi C). , 1998, Hasegawa H, 1995, Lee SJ, 1999), anti-allergic effects (Bae EA, Choo MK, 2003, Kim DH, 2003, Lee JH, 2006). Several studies have shown that some ginsenosides exhibit glucocorticoid-like activity, which directly or indirectly activates the glucocorticoid receptor (GR) (Lee YJ, 1997, Lee YN, 1996). Other studies have suggested a novel treatment for inflammatory diseases by harmonizing control between homeostasis and immune action by interactions between toll like receptor 4 (TLR4) and GR (Ogawa S, 2005).

The present invention seeks to explain the potential of a novel therapeutic agent to treat fatal sepsis by discovering the fact that Compound K modulates the response caused by endotoxin. It was found that pretreatment and posttreatment of Compound K resulted in the recovery of survival of sepsis-induced mice caused by LPS of Gram-negative bacteria, and that Compound K regulates systemic inflammatory responses in vivo. Therefore, the present invention confirmed that the compound K shows the effects of anti-inflammatory and anti-septicemia.

Accordingly, the present inventors have made intensive studies to overcome the problems of the prior art, and as a result, the compound K (Compound K), which is a red ginseng saponin convertor, confirmed that the antiseptic effect was excellent in sepsis and sepsis shock caused by Gram-negative bacteria, The present invention was completed.

Therefore, the main object of the present invention is to provide a composition containing Compound K (Compound K) which is a red ginseng saponin convertor having antiseptic efficacy as an active ingredient.

According to one embodiment of the present invention, the present invention provides a composition for treating sepsis, which contains Compound K (Compound K) represented by the following Chemical Formula 1 as an active ingredient. In the present invention, a novel use of compound K, known as the cinnanoside metabolite, known in the art, namely, the anti-septic effect of compound K was disclosed.

[Formula 1]

In the composition of the present invention, the compound K is characterized in that the red ginseng saponin ginsenosides Rb1, Rb2, Rc or a conversion from Rd. Compound K of the present invention is a substance not found in red ginseng, and is a saponin convertor in which red ginseng saponin is converted by enterobacteriaceae. In the examples of the present invention, compound K can be obtained by incubating protophanaxadiol type saponins (ginsenosides Rb1, Rb2, Rc and Rd) with Aspergillus niger derived cellulase. While the efficacy of known compounds K is primarily an anticancer effect, the efficacy identified in the present invention is anti sepsis, which can be used as a pharmaceutical composition.

In the composition of the present invention, the sepsis is characterized in that it is preferably caused by LPS of Gram-negative bacteria. In an embodiment of the present invention, Gram-negative sepsis caused by Gram-negative bacteria was induced by LPS of Gram-negative bacteria, which is a form of sepsis, and then the compound K of the present invention was administered to confirm the anti-septic effect.

In order to use the composition of the present invention as a pharmaceutical composition can be prepared by a method known in the pharmaceutical field, powders, granules, tablets, capsules by themselves or mixed with a pharmaceutically acceptable carrier, excipient, diluent, etc. Or in the form of an injection or the like. They can also be administered orally or parenterally.

The dosage for administering Compound K according to the present invention as an active ingredient may be appropriately selected according to the age, sex, condition, and symptoms of the disease of the patient, and preferably 0.01 to 100 mg / kg (body weight) of compound per day. K may be administered.

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

As described above, according to the present invention, compound K, which is a metabolite of ginsanoside, which is red ginseng saponin, can be obtained by using an enzyme, and this compound K is easily absorbed by humans and its stability has already been verified. The present invention confirms the excellent anti-septic effect in sepsis-induced mice as a new use of compound K, suggesting the possibility of use as a pharmaceutical composition for the treatment of sepsis.

Example 1. Preparation and Separation of Compound K

Compound K, one of the ginsenoside metabolites, was isolated as follows. 1 g of protopanaxadiol-type saponin (ginsenoside Rb1, Rb2, Rc and Rd, self-separated from KT & G Central Research Institute) was incubated with Aspergillus niger derived cellulase (cellulase) for 48 hours at 37 ° C. The reaction was extracted with n-butanol, the butanol fraction was concentrated, the compound K was separated from the silica gel column using chloroform-methanol (9: 1) as the mobile phase and chromatographed again using an ODS (C18) column. Was performed. Purification was analyzed by HPLC (Waters HPLC system): column, Discovery C ₁₈ (25 × 0.46 cm, 5um, Supelco); elution solvent, solvent A (water) and solvent B (acetonitrile) -gradient profile of solvent A to solvent B; from 80: 20 to 10: 90 for 0-90 min .; Detector, UV 203 nm. The separated compound K was identified using ¹³ C-NMR, ¹ H-NMR, HPLC, FAB-MS, the purity of the compound K was more than 97%.

Example 2 Preparation of Experimental Animals and Materials

All experiments described herein were performed using mice with a C57BL / 6 background. All animal-related procedures were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee of Chungnam National University. Mice were used for the LPS test at 8-10 weeks of age. Test groups were classified by age and gender. Escherichia coli O26: B6 LPS (Sigma-Aldrich) was diluted in sterile PBS and injected into animals intraperitoneally.

Example 3 Efficacy Analysis of Compound K (C-K) Using Sepsis Mouse Model

To determine whether C-K (Compound K) is effective in endotoxin-induced sepsis in vivo, a murine sepsis model was constructed. CK (10, 30 and 50 mg / kg) was orally administered to C57B / L6 mice (25 per group). One day later, gram-negative bacterial endotoxin (LPS) was injected into the abdominal cavity by 40 mg / kg (body weight) to cause septic shock. The survival rate was then examined for 5 days.

LPS at 40 mg / kg caused 90% death of the control group, but LPS-injected mice increased the survival rate concentration-dependently when C-K was orally administered. 85% survived in the highest concentration test group (50 mg / kg) and 50% survived even at 30 mg / kg (P <0.05) (FIG. 2).

Example 4 Inhibitory Effects of C-K on Endotoxin-Derived Lethal Shock

It was investigated whether C-K can protect mice from endotoxin shock caused by LPS. Mice were divided into groups and injected intraperitoneally (i.p.) at an LPS concentration of 40 mg / kg. Injecting LPS at 40 mg / kg body weight resulted in 100% death within 40 hours in the control group (n = 25). At this time, when C-K was administered after 6 hours, it was observed that 70% survived for 100 hours (FIG. 3A). In the other group, the LPS administered at 30 mg / kg body weight was 70% mortality in the control group, but significantly increased survival when treated with C-K (FIG. 3B) (P <0.006). These results suggest that C-K may be a novel therapeutic for the treatment of gram-negative septic shock.

1 is a chemical formula of compound K of the present invention.

Figure 2 shows the effect of Compound K of the present invention on sepsis caused by endotoxin in vivo.

Figure 3 shows the inhibitory effect of endotoxin-derived lethal shock of compound K of the present invention.

Claims (4)

Sepsis composition for treating sepsis containing a compound K (Compound K) represented by the formula (1) as an active ingredient. The composition of claim 1, wherein the compound K is a convertor of ginsenosides Rb1, Rb2, Rc or Rd which is red ginseng saponin. The composition of claim 2, wherein the compound K is prepared by digesting the ginsenoside with Aspergillus niger derived cellulase. The composition of claim 1, wherein the sepsis is caused by Lipopolysaccharide (LPS) of Gram-negative bacteria.

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