JPH04243833A - 11beta-hydroxysteroid dehydrogenase ingibitor - Google Patents

Abstract

PURPOSE:To provide a compound having 11beta-hydroxysteroid dehydrogenase- inhibiting activity and useful as medicines such as antiinflammatory agent and tonic. CONSTITUTION:The objective 11beta-hydroxysteroid dehydrogenase inhibitor contains Panax ginseng, its extract or ginsenoside Rd as an active component.

Description

[Detailed description of the invention]

0001

[Industrial Application Field] The present invention has an 11β-hydroxysteroid dehydrogenase inhibitory effect and is an anti-inflammatory agent.
It relates to compounds useful as pharmaceuticals such as air supplements.

0002

BACKGROUND OF THE INVENTION Among the anti-inflammatory drugs currently in clinical use, there is a series of compounds generally referred to as steroidal anti-inflammatory drugs. They are steroid compounds secreted from the adrenal cortex or chemically modified versions thereof, and most of them have oxygen at the C-11 position, ie, 11-hydroxysteroids.

Specific examples include cortisone, hydrocortisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, and the like.

[0004] Oxygen at position 11 is thought to be important for the action of steroid anti-inflammatory drugs, and 11β-hydroxysteroid dehydrogenase is an enzyme that reduces oxygen at position 11. because there is,
Drugs that inhibit the function of this enzyme are considered to be useful as anti-inflammatory drugs.

[0005]

Problem to be Solved by the Invention The present inventors have discovered that 11β
- As a result of intensive research to find a compound that has a hydroxysteroid dehydrogenase inhibitory effect, we found that the herbal medicine ginseng, its extract, and one of its constituents, ginsenoside Rd, which has been frequently used in Chinese medicine since ancient times, has this effect. This discovery led to the completion of the present invention.

That is, the present invention is an 11β-hydroxysteroid dehydrogenase inhibitor containing ginseng, an extract thereof, or ginsenoside Rd as an active ingredient.

Hereinafter, ginseng, its extract, and ginsenoside Rd will be collectively referred to as the active ingredients of the drug of the present invention.

[0008] Ginseng is one of the crude drugs frequently used in Chinese medicine, and is used for tonicity, tonicity, fertilization, sedation, etc. Ginsenoside Rd is a saponin isolated from the herbal medicine ginseng.

It was not previously known that the active ingredient of the drug of the present invention has an 11β-hydroxysteroid dehydrogenase inhibitory effect, and this was revealed for the first time by the present inventors.

[0010] The ginseng extract as used herein refers to an extract obtained by extracting ginseng with water and/or an organic solvent and distilling off the extraction solvent from the extract.

[0011] When extracting with water, 10 to 20
It is desirable to add twice the amount and extract at 90 to 100°C for about 30 to 90 minutes.

[0012] Specific examples of organic solvents in the case of extraction with organic solvents include alcohols, acetone, etc., and examples of alcohols include methanol, ethanol, propanol, butanol, etc. The extraction temperature may be room temperature, but in consideration of extraction efficiency, it may be heated to around the boiling point of the solvent used. Further, the extract can be separated by methods such as decantation and centrifugation.

[0013] Ginsenoside Rd can be obtained, for example, as follows.

The ginseng extract obtained as described above is defatted using ether or the like, and then extracted again using alcohol (same as above) or an organic solvent such as chloroform, ether, or ethyl acetate.

[0015] The extract obtained by extraction is used as it is or dissolved in a solvent, and one or more of water, alcohol such as methanol, organic solvent such as chloroform, ether, and ethyl acetate is used as a developing solvent, and silica gel or the like is used as a carrier. It can be obtained by subjecting it to column chromatography or high performance liquid chromatography one or more times.

Next, ginseng extract and ginsenoside R
A specific example of the production of d is shown below.

Specific example 1 Add 1 liter of water to 100 g of carrots,
After heating and extracting at 5° C. for 60 minutes and cooling to room temperature, the filtered extract was dried to obtain 43 g of dry extract powder.

Specific example 2 50g of carrots and 500ml of ethanol
1 was added and extracted, and the solvent of the filtered extract was distilled off to obtain 18 g of dry extract powder.

Specific Example 3 Carrots were cut into small pieces, extracted with methanol, defatted with ether, and further extracted with n-butanol. The solvent was distilled off, and 65 g of the obtained n-butanol extract was subjected to silica gel column chromatography and divided into seven fractions from I to VII using chloroform:methanol:water (7:3:0.5) as a developing solvent. Ta.

Fraction III was again subjected to silica gel column chromatography using chloroform:methanol:water (7:2:0.5) as a developing solvent.
It was divided into three fractions: II-1, III-2 and III-3.

Fractions IV and V were again subjected to silica gel column chromatography using chloroform:methanol:water (7:3:0.5) as a developing solvent, and five fractions from IV・V-1 to V・V-5 were separated. divided into fractions.

Regarding the III-3 fraction (4.1 g) and the IV/V-2 fraction (6.84 g),
High performance liquid chromatography using 50% methanol and then 75% methanol as a developing solvent [Column: Inert
sil Prep ODS (20mmφ×250m
m, 10 μm), flow rate: 15 ml/min, column temperature: room temperature, detection: 203 nm], and by distilling off the solvent of the 75% methanol eluate, ginsenoside Rd0.
75g was obtained.

Next, the active ingredient of the drug of the present invention is 11β-
The fact that it has a hydroxysteroid dehydrogenase inhibitory effect will be explained using experimental examples.

Experimental example Sprague Daw
ley) strain male rats (250-350 g) using the method of Remmer et al.
im H, Schenkman JB, Estab
rook RW (1967) Methods
of the elevation of
hepatic microsomal mix
ed function oxidase le
vels and cytochrome P-
450. In: Fleischer S, Est.
abrook RW (eds) Methods
in enzymelogy, vol 10. A
Academic Press, New York,
Microsomes were obtained according to [pp703-708]. That is, the above kidney was transferred to an ice-cold Tris buffer containing 0.25M sucrose solution, and then added to an STE solution (0.25M sucrose solution).
25M, Tris buffer 10mM, magnesium chloride 2m
M, dithioerythritol 1mM).
Centrifuge the supernatant at 0xg for 10 minutes at 8,000xg for 10 minutes.
min, 15,000 x g for 10 min, 105,000 x
The residue obtained by centrifugation at 105,000 x g for 60 minutes was dissolved in 0.15 M potassium chloride solution, and microsomes were obtained by centrifuging at 105,000 x g for 30 minutes.

[0025] 95% of the obtained rat kidney microsomes
Substrate (Krebs-Henseleit solution) containing oxygen and 5% oxygen
Test solutions (1 to 1.5 ml) adjusted to various concentrations using NAD+, NADH, NADP+, NADPH)
and tritium (3H)-labeled corticosterone (1,2,6,7-3H-Corticosterone
ne, 80 Ci/mol) and gently incubated in a 37°C water bath for 15-60 minutes. This was further extracted with ether, and after distilling off the ether, it was dissolved in 100 μl of methanol/water (55/45).
The 11β-hydroxysteroid dehydrogenase inhibitory effect of the active ingredient of the drug of the present invention was investigated by subjecting μl to reverse-phase high performance liquid column chromatography and measuring labeled corticosterone using a UV detector. Table 1 shows the 11β-hydroxysteroid dehydrogenase inhibitory effect of the active ingredient of the drug of the present invention as the inhibition rate (%) when compared with the control group.

Table 1

From the above results, the 11β-hydroxysteroid dehydrogenase inhibitory effect of the active ingredient of the drug of the present invention was confirmed.

[0028] Furthermore, as a result of an acute toxicity test of the active ingredient of the drug of the present invention using ICR rats, there was no death even after oral administration of 1 g/kg, confirming that the compound is highly safe. It was done.

[0029] Therefore, the active ingredient of the drug of the present invention is useful as a medicine such as an anti-inflammatory agent, that is, a medicine for the prevention and treatment of diseases that can be expected to be effective by inhibiting 11β-hydroxysteroid dehydrogenase.

Next, the dosage and formulation of the active ingredient of the drug of the present invention will be explained.

The active ingredient of the drug of the present invention can be administered to animals and humans as is or together with a conventional pharmaceutical carrier. The dosage form is not particularly limited and may be selected and used as necessary, and may include oral dosage forms such as tablets, capsules, granules, fine granules, and powders, and parenteral dosage forms such as injections and suppositories. It will be done.

[0032] In order to exert the desired effect as an oral preparation, the active ingredient of the drug of the present invention for an adult is usually 30 mg, depending on the age, weight, and severity of the disease of the patient.
It seems appropriate to take ~2g in divided doses several times a day.

[0033] In the present invention, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods using, for example, starch, lactose, sucrose, mannitol, carboxymethyl cellulose, cornstarch, inorganic salts, and the like. In addition to the above-mentioned excipients, this type of preparation may include binders, disintegrants,
Surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents,
Flavors etc. can be used. Specific examples of each are shown below.

[Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch,
Methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low substituted hydroxypropyl cellulose.

[Surfactant] Sodium lauryl sulfate,
Soy lecithin, sucrose fatty acid ester, polysorbate 80.

[Lubricant] Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The active ingredient of the drug of the present invention can also be administered as a suspension, emulsion, syrup, or elixir, and these various dosage forms may contain flavorings and coloring agents. You may.

[0040] In order to exert the desired effect as a parenteral agent, it depends on the age, weight, and severity of the disease of the patient;
Normally, as an adult, the weight of the active ingredient of the drug of the present invention is 0 per day.
．． Intravenous injections, intravenous drips, subcutaneous injections, and intramuscular injections of 1 to 50 mg are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and diluents generally include distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, and polyethylene glycol. etc. can be used. Furthermore, a bactericide, a preservative, and a stabilizer may be added as necessary. In addition, from the viewpoint of stability, this parenteral preparation may be filled into a vial or the like, then frozen, water removed by ordinary freeze-drying techniques, and a liquid preparation prepared from the freeze-dried product immediately before use. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate.

Other parenteral preparations include liquid preparations for external use, liniments such as ointments, suppositories for intrarectal administration, etc.
Manufactured according to conventional methods.

The present invention will be explained in more detail below with reference to Examples, but the present invention is not limited thereto in any way.

Example 1 According to the above recipe, ① to ② were mixed uniformly and compressed using a tablet machine to obtain tablets each weighing 200 mg. One tablet contains 20 mg of ginseng extract, and adults should take 5 to 15 tablets a day in several doses.

Example 2 According to the above recipe, ■, ■, and part of ■ were uniformly mixed, compressed and molded, and then pulverized, the remaining amounts of ■ and ■ were added, mixed, and compressed using a tablet machine. Molded tablet 200mg
tablets were obtained. This tablet contains ginsenoside Rd2
It contains 0mg, and adults should take 5 to 15 tablets a day in several doses.

Example 3 ①, ② and ② were uniformly mixed according to the above-mentioned recipe, and the mixture was homogenized by a conventional method, and granulated using an extrusion granulator.
After drying and crushing, (1) and (2) were mixed and compressed using a tablet machine to obtain tablets each weighing 200 mg. One tablet of this tablet contains 20 mg of ginsenoside Rd, and adults should take 5 to 15 tablets a day in several doses.

Example 4 [0047] Items (1) to (4) were uniformly mixed according to the above recipe, compression molded using a compression molding machine, crushed using a crusher, and sieved to obtain granules. 1 g of this granule contains 100 m of carrot extract.
It contains 1 to 3 g per day for adults, divided into several doses.

Example 5 According to the above recipe, ① to ② were uniformly mixed and made into a paste. After granulation using an extrusion granulator, the mixture was dried and sieved to obtain granules. 1 g of this granule contains ginsenoside Rd
It contains 100mg, and adults should take 1 to 3g in several doses per day.

Example 6 ■~■ were mixed uniformly according to the above recipe, and 200
mg was filled into No. 2 capsules. One capsule of this preparation contains 20 mg of ginsenoside Rd, and adults should take 5 to 15 capsules a day in several doses.

Example 7 According to the above recipe, ■ was dissolved in ■ and ■, and the solutions of ■ and ■ were added to emulsify to obtain an injection.

Claims (2)

[Claims] Claim 1: 1 containing ginseng or its extract as an active ingredient
1β-hydroxysteroid dehydrogenase inhibitor. 2. An 11β-hydroxysteroid dehydrogenase inhibitor, wherein the active ingredient is ginsenoside Rd.