KR20080058525A - Pharmaceutical composition, health food composition, cosmetic composition and farnesoid x-activated receptor inhibitors, containing sterols derivatives - Google Patents

Abstract

A sterol derivative is provided to lower cholesterol level in liver by promoting catabolism of the cholesterol into bile through inhibition of the activity of farnesoid X-activated receptor(FXR), thereby being useful for treating and preventing metabolic diseases and hyperlipidemia. A pharmaceutical composition for preventing or treating diabetes, obesity, hypercholesterinemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, cardiovascular disease, compulsive overeating and ischemic heart diseases or for inhibiting the activity of a farnesoid X-activated receptor comprises a sterol derivative represented by a formula(1), wherein the sterol derivative is extracted from Stelodoryx chlorophylla, one of the porifera. In the formula(1), R1 is a group 1 or a group 2.

Description

Pharmaceutical composition, health food composition, cosmetic composition and farnesoid X-activated receptor inhibitors, containing sterols derivatives}

The present invention is a pharmaceutical composition for the prevention or treatment of diabetes mellitus, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, overeating, ischemic heart disease using the sterol derivative of the formula (1) as an active ingredient The present invention relates to a composition, a health food composition and a cosmetic composition, and to a farnesoid X-activated receptor activity inhibitor composition, wherein the sterol derivative compound of formula 1 is a marine natural product purified from sponge animals.

[Formula 1]

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to be.]

Excess cholesterol accumulation in the body can lead to fatal diseases such as neurodegenerative diseases as well as cardiovascular diseases [ PNAS , 1999 , 96, 6660]. Therefore, bile acids produced from cholesterol in the liver are excessively accumulated in the body to remove toxic cholesterol and help absorb fat-soluble essential vitamins. Farnesoid X-activated receptors (FXRs), known as nuclear receptors involved in human lipid and cholesterol metabolism, are central factors in maintaining homeostasis in the production of bile in the body. Activation of the panasoid X nuclear receptors inhibits the production of cholesterol 7 alpha-hydroxylase, the most important enzyme for the catabolism of cholesterol to bile bile [ Science , 1999 , 284, 1362; Science , 2001 , 294, 1866]. Therefore, inhibiting the activity of the panasoid X nuclear receptor promotes the production of bile, which is useful for removing excessively accumulated cholesterol in vivo. For example, guggulsterone, an antihyperlipidemic drug used in India for 2,000 years, acts as an antagonist of the panasoid X nuclear receptor, and has been reported in the literature to effectively lower liver cholesterol levels [ Science , 2002 , 296, 1703. Thus, antagonists selective to the panasoid x nuclear receptor can be developed for the treatment of cholesterol metabolic diseases and hyperlipidemia.

The sterol derivative compound of Formula 1 has been reported to be isolated from Stelodoryx chlorophylla , which is a sponge animal [ J. Nat . Prod . 1993 , 56, 282], the physiological activity of the antagonistic effect of the nuclear receptor FXR is unknown.

An object of the present invention is to prevent or treat diabetes mellitus, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, overeating, ischemic heart disease as an active ingredient comprising a sterol derivative compound as an active ingredient. To provide a pharmaceutical composition, health food composition and cosmetic composition, another object of the present invention to provide a farnesoid X-activated receptor activity inhibitor composition comprising a sterol derivative compound as an active ingredient will be.

The present invention is a pharmaceutical composition for the prevention or treatment of diabetes mellitus, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, overeating, ischemic heart disease using the sterol derivative of the formula (1) as an active ingredient A composition, a health food composition and a cosmetic composition.

In addition, the present invention is a major factor involved in cholesterol metabolism in humans, when the activity is inhibited by promoting the catabolism of cholesterol from the bile to effectively lower the cholesterol level in the liver (farnesoid X-activated receptor; FXR ), The present invention relates to an activity inhibitor composition.

[Formula 1]

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to be.]

The sterol derivative compound of Chemical Formula 1 is a marine natural product purified from sponge animals, and the sterol derivative of Chemical Formula 1 is 3β-hydroxy-17β-pregin-5-en-20-one (3β-Hydroxy-17β). -pregn-5-en-20- one) and (22 E) -3β- hydroxy -26,27- bis play Collet star -5,22- diene -24- one ((22 E) -3β-Hydroxy- 26,27-bisnorcholesta-5,22-dien-24-one) and refer to the following compound (1) and compound (2), respectively.

The dosage of the pharmaceutical composition and the farnesoid X-activated receptor active inhibitor composition used to achieve the therapeutic effect according to the invention, as well as the specific compound, the method of administration, the subject to be treated and the disease to be treated However, based on the sterol derivative compound of Formula 1, which is the active compound according to the present invention, it is usually about 0.001 mg / Kg to 400 mg / Kg, and may be administered once or several times a day, and the dosage is the weight of the patient, Age, sex, health status, diet, administration time, administration method, excretion rate and the severity of the disease can be adjusted in various ways, the administration method is a tablet, capsule, powder form, solution and suppository form that can be administered orally as oral Or parenteral administration (eg intravenous subcutaneous, intraperitoneal, topical or visual route) but oral administration is more preferred.

Oral administration of the pharmaceutical composition according to the present invention can be prepared in any of a variety of existing forms, for example tablets, powders, dry syrups, chewable tablets, granules, chewing tablets, capsules, soft capsules, pills, It can exist in various forms, such as drink, sublingual tablet, and the like. In the case of powder, the amount of the active ingredient is preferably 0.01 to 99.9% by weight, and the content is applied in a rational manner according to the formulation of the composition. The pharmaceutical composition according to the present invention may be difficult to maintain physical properties when the amount of the pharmaceutically acceptable salt of the sterol derivative compound according to each formulation exceeds the maximum total weight, and less than the minimum weight. The effect may not be sufficient.

Tablets according to the present invention may be administered to a patient in any form or manner in which an effective amount is bioavailable, ie, by oral route, and is suitable for administration depending on the nature of the disease state to be treated, the stage of the disease, and other relevant circumstances. The form or manner can be easily selected and the composition according to the invention can comprise one or more pharmaceutically acceptable excipients, wherein the proportions and properties of such excipients are dependent on the solubility and chemical properties of the selected tablets, the chosen administration Determined by the route and standard pharmaceutical practice.

More specifically, the compositions according to the invention may comprise a therapeutically effective amount of the above-mentioned active ingredient as an essential ingredient together with one or more pharmaceutically acceptable excipients. Excipient materials can be solid or semisolid materials that can function as a vehicle or medium of the active ingredient, and suitable excipients are well known in the art. Excipient materials may be selected in connection with the intended dosage form, specifically for tablets, powders, chewable tablets, granules, chewing tablets, capsules, soft capsules, pills, sublingual tablets or syrups, The therapeutically active drug component may be combined with any oral nontoxic pharmaceutically acceptable inert excipient such as lactose or starch. Optionally, the pharmaceutical tablets of the present invention may contain a binder such as amorphous cellulose, gum tragacanth or gelatin, a disintegrant such as alginic acid, a lubricant such as magnesium stearate, a glidant such as colloidal silicon dioxide, sucrose It may also contain sweetening agents such as oz or saccharin, colorants or flavoring agents such as peppermint or methyl salicylate.

Because of their ease of administration, tablets may be the most advantageous oral unit dosage form, and if desired, tablets may be coated with sugar, shellac or other enteric coatings by standard aqueous or non-aqueous techniques, respectively. Tablets or capsules preferably contain about 10 mg to 200 mg of active ingredient.

Metabolic disorders such as diabetes, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, hyperphagia, ischemic heart disease using the sterol derivative compound of Formula 1 as an active ingredient It is useful as a health food composition for the prevention of. The health food supplement of the present invention using the sterol derivative compound as an active ingredient is useful for preventing stroke and may assist in the treatment of related diseases.

In one embodiment, as an essential ingredient of the health food composition is not particularly limited to ingredients added in addition to the sterol derivative compound of the present invention, when the composition of the present invention is used as a food or beverage additive, the composition is added as it is or other It can be used together with a food or food ingredient, or can be suitably used according to a conventional method.

At this time, the extract in the food or beverage, that is, in general, the health food composition of the present invention can be added to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.001 to 10 g, preferably based on 100 ml Can be added in a ratio of 0.3 to 1 g. The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the extract as an essential ingredient in the ratios indicated, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract, for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. . The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components may be used independently or in combination, and the proportion of the additive is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail.

Example 1 Isolation and Purification of the Sterol Derivative Compound (1) and Compound (2)

The extraction method of the sterol derivative compound (1) and compound (2) according to the present invention is as follows. After thawing the frozen animals stored in the sea and thawed, and then finely chopped the sponges to prepare a sample, the samples were mixed three times, each of 1: 1 ratio methanol: dichloromethane (CH ₂ Cl ₂ ) mixed organic Extracted at room temperature with a solvent. The extract was then vaporized under reduced pressure to afford a residue, which was first partitioned extracted with methanol and normal-hexane. Then, the extract of the solvent, methanol, was evaporated under reduced pressure to obtain a residue, and the residue was secondly extracted with water and ethyl acetate.

The ethyl acetate extract was vaporized under reduced pressure to obtain a residue. This residue was chromatographed on a silica column, using hexane and ethyl acetate as the eluting fluid. Fractions collected as above were subjected to C-18 reversed phase semi-preparative HPLC column, 10 μm, 250 × 10 mm, UV detector = 210 nm, elution rate 2.5 ml chromatographed with acetonitrile: water (60:40) as eluent on the eluent to obtain an eluate, and the eluate was vaporized under reduced pressure to give a sterol-based compound in the amorphous form (1) And compound (2) were obtained, respectively. The retention times on the chromatography of compounds (1) and (2) were 30 and 50 minutes, respectively.

Example 2 Determination of Chemical Structure of Sterol Derivative Compound (1) and Compound (2)

Identification process of the amorphous pure compound (1) and the compound (2) obtained in Example 1 was carried out by ¹ H-nmr (nuclear magnetic resonance), ¹³ C-nmr, LRMS and HRMS experiments This was done by analyzing the results and two-dimensional NMR data. That is, nmr experiments were conducted with respect to Compounds (1) and (2), and the structures of Compounds (1) and (2) were determined therefrom. System derivatives, marine naturals already reported in the literature [ J. Nat . Prod . 1993 , 56, 282.

Compound (1) ¹ H NMR (600 MHz, CDCl ₃ ) δ 5.35 (m, 1H), 3.53 (m, 1H), 2.53 (t, 1H, J = 11.2 Hz), 2.30-2.23 (m, 3H), 2.12 (s, 3H), 2.06-2.04 (m, 2H ), 1.87-1.84 (m, 2H), 1.68-1.44 (m, 8H), 1.24-1.09 (m, 3H), 1.01 (s, 3H), 0.99 (m, 1H), 0.88 (m 1H), 0.63 (s, 3H)

Compound (2) ¹ H NMR (600 MHz, CDCl ₃ ) δ 6.64 (dd, 1H, J = 15.8, 8.8 Hz), δ 5.99 (d, 1H, J = 15.8 Hz), 5.34 (m, 1H), 3.52 (m, 1H), 2.30-2.25 (m, 2H) , 2.23 (s, 3H), 2.01-1.95 (m, 2H), 1.85-1.83 (m, 2H). 1.66-1.53 (m, 12H), 1.28-1.27 (m, 2H), 1.10 (d, 3H, J = 6.5 Hz), 1.08 (m, 1H), 1.01 (s, 3H), 0.96 (dt, 1H, J = 11.8, 4.6 Hz), 0.73 (s, 3H)

Example 3 Antagonistic Effects of Sterol Derivative Compounds (1) and Compounds (2) on the Panasonic X Nuclear Receptor ( FXR )

FXR antagonistic effects of the amorphous pure substance sterol derivative compound (1) and compound (2) were assayed by Cotransfection assay. Here, the cotransfection assay refers to animal cells cultured in DMEM medium containing 10% bovine serum and 1% antibiotic under a 5% carbon dioxide environment for one day, and then to the FXR protein expression gene and FXR, which are human nuclear receptors. After injecting and expressing a vector containing a luminescent gene regulated by the cell, the cultivation was performed for one more day by adding CDCA (chenodeoxycholic acid), an extract of FXR and an extract or natural product, and confirming the expression level of the luminescent gene. As a general method of comparing the degree of inhibition with that of the control group ( Z- guggulsterone), the beta galactosidase gene was used to correct an error in pipetting during sample addition.

FXR antagonistic effect of the sterol derivative compound (1) and compound (2) was shown in Table 1. The sterol derivative compounds (1) and compound (2) have no cytotoxicity and show excellent FXR antagonistic effects.

Table 1 FXR antagonist effect and cytotoxicity of sterol derivative compound (1) and compound (2)

From the above results, it can be seen that the sterol derivative compound (1) and compound (2) are useful compounds for treating metabolic diseases and hyperlipidemia as new FXR antagonists.

The sterol derivative compound according to the present invention inhibits the activity of farnesoid X-activated receptor (FXR), promotes catabolism from cholesterol to bile, and effectively lowers cholesterol levels in the liver, thereby reducing metabolic diseases and hyperlipidemia. It is useful for treatment and prevention.

Claims (5)

A pharmaceutical composition for preventing or treating diabetes mellitus, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, hyperephagia, and ischemic heart disease using the sterol derivative of Formula 1 as an active ingredient. [Formula 1]

[R ₁ is

or

to be.] The method of claim 1, The sterol derivative is a pharmaceutical composition for the prevention or treatment of diabetes, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, overeating, ischemic heart disease, characterized in that it is extracted from a sponge animal . A health food composition for preventing or treating diabetes mellitus, obesity, hypercholesterolemia, hyperlipoproteinemia, hyperlipidemia, arteriosclerosis, hypertension, circulatory disease, overeating, ischemic heart disease using the sterol derivative of Formula 1 as an active ingredient. [Formula 1]

[R ₁ is

or

to be.] Cosmetic composition for the prevention of obesity or to improve obesity using a sterol derivative of the formula (1) as an active ingredient. [Formula 1]

[R ₁ is

or

to be.] A farnesoid X-activated receptor activity inhibitor composition comprising the sterol derivative of Formula 1 as an active ingredient. [Formula 1]

[R ₁ is

or

to be.]