WO1994002160A1 - Tonic - Google Patents

Abstract

Muirapuama is the root of a bush of the family Olacaceae, which grows in the Amazon River basin, Brazil, and the original vegetation of which includes Ptychopetalum olacoides, P. uncinatum and Liriosma ovata. Its extract has been known from of old as a tonic and aphrodisiac, but it has problems of, for example, low drug efficacy. Now the present inventors have found that the extract containing at least 0.033 % of β-sitosterol, prepared by condensing a solution obtained by macerating a chopped crude drug containing muirapuama with 50-99 % ethanol or another solution obtained by further macerating the residue of the above solution with 50-99 % ethanol, has more potent tonic and aphrodisiac effects than those of the conventional extracts and dried extracts. Further it has been found that drugs containing the extract are efficacious in preventing or treating impotency, subvirility, and the like caused by stress.

Description

 Description Tonics Technical field

 The present invention relates to tonic extract. Background art

 Mirabuama is the root of a shrub of the family Boloniaceae (olcaeceae) that grows in Brazil and the Amazon River basin.

Unicinatum, Liriosmauovata, 6 ', and these exotics have long been known as tonic aphrodisiacs.

 Conventionally, extracts extracted from Mui Lapuama have had problems such as low efficacy.

Disclosure of the invention

 The present inventors have considered that mui lapuama (Ptychhopetalum)

olacoides) with 50% to 99% ethanol 3 times the volume of the extract, and the extract and the extraction residue were further immersed and extracted with a 50% to 99% ethanol 3 times the volume. It has been found that the tonic aphrodisiac effect is higher than that of the dried extract of the present invention, and the present invention has been completed based on this finding.

 That is, according to the present invention, a liquid obtained by immersing a crude drug shred of muirapuama with 50% to 99% ethanol and a liquid obtained by immersing a residue of the immersion liquid with 50% to 99% ethanol are concentrated; It is an extract containing 0.033% or more of sitosterol.

The extract of the present invention can be produced by the production method described below. -l-That is, muirapuama (P tychopeta 1 um

o 1 ac o i d e s) Extract the shredded pieces (20000 m or less) with 50% to 99% ethanol 3 times for about 2 hours and filter the leachate. This filtrate is referred to as filtrate (a). Further, the residue is leached for about 2 hours with 50% to 99% ethanol three times the volume, and the leaching solution is filtered. This filtrate is referred to as filtrate (b). The filtrate (a) and the filtrate (b) are concentrated under reduced pressure to obtain a residue. The resinous substance adhering during the concentration under reduced pressure is thoroughly dissolved in 99% ethanol, and the residue is combined with the residue to prepare a 5% extract yield.

 The property of the extract of the present invention is a brown liquid, which is characterized by having a high fat-soluble component content.

 The kiss of the present invention is preferably in the range of 0.2 to 3.0 gZ50 Kg in terms of a crude drug, particularly preferably 0.2 to 1.55 Kg, as a dose to an adult. preferable. The extract of the present invention is prepared into liquid preparations such as solutions, suspensions and emulsions or solid preparations such as granules (powder, granules), capsule preparations and tablets by ordinary methods, and used as oral preparations Can be provided.

 In shaping these dosage forms, those conventionally known in the art can be used as the carrier, and they are produced by means commonly used in the art.

 Examples of diluents that can be used for preparing liquid preparations for oral administration include water, ethanol, glycerol, propylene glycol, agar, and tragacanth, and other surfactants, coloring agents, and flavoring agents as necessary. Can be used.

Examples of excipients that can be used in the preparation of solid preparations for oral administration include, for example, excipients usually used in oral preparations (eg, milk powder, ¾5 hammer, mannitol, crystal cell D-source), disintegrants (horse Bell powder, corn starch, low-substituted hydroxypropylcellulose, carboxymethylcellulose, etc. Polyalcohol, hydroxypropyl bilcellulose, tannin castor oil, etc.), and if necessary, flavors, additives, preservatives, etc. can be added. BEST MODE FOR CARRYING OUT THE INVENTION

 Next, the present invention will be described in more detail with reference to Examples and Test Examples.

Example 1

 Mui Lapuama (Ptychopeta 1 umo 1 acoides) Tagiri (200 m or less) 3 80 Kg is soaked and extracted with 70% ethanol 3 times (114 L) for 2 days, and the leachate is extracted with 1 2 The mixture was filtered through a 0-mesh stainless steel wire mesh. 700 L of this filtrate was used as filtrate (a). Further, the residue is immersed in a beach for 2 days with 70% ethanol (3 times the amount of ethanol) (110 L), and the leachate is filtered through a 120 mesh stainless steel wire mesh. The filtrate (1200 L) was used as filtrate (b). The filtrate (a) and the filtrate (b) are combined and concentrated under reduced pressure at 150 mmHg to obtain 16 kg of a residue. The resinous substance adhering to the container during the concentration under reduced pressure was thoroughly dissolved in 40 L of 99% ethanol, and the residue was combined with the residue to obtain 19 kg (yield: 5.2%) of the concentrated extract under reduced pressure.

 The resulting extract was a brown liquid with a peculiar smell, and the taste was somewhat astringent and astringent.

The production of Example 1 was repeated seven times (lots 1 to 7), and the obtained extracts were heated in a water bath at 50 ° C. for 30 minutes, mixed well, and 5 g of the mixture was precisely weighed. And placed in a 50 ml stoppered centrifugal sedimentation tube. To this, 5 ml of methanol and 5 ml of water were added, 20 ml of hexane was added three times, and the mixture was shaken for 10 minutes each time, followed by centrifugation at 300 rpm for 5 minutes. . The supernatant was separated and passed through a funnel in which 5 g of anhydrous sodium sulfate was placed on a small amount of absorbent cotton. The solvent was distilled off under reduced pressure on a water bath, and 5 ml of an internal standard solution was accurately added to the residue to dissolve it, and used as a standard solution. The sample solution (T) and the standard solution (S) were each taken at 1; z 1 each, and the test was carried out by gas chromatography under the following conditions. To the peak Thus ¾ of internal standard ¾1 material of each liquid; 5 - to则定ratio Qt and Q _s of peak Thus i of collected by sterols. Contains citosterol (C29II500) of this product — (%)

 Qt 1 0 0

 Constant CtfflyS—Sitosterol (mg) X X

Q _s sample collection ttHmg) Internal standard solution; testosterone propionate in chloroform solution (3 → 2000)

Operating conditions

 Column; DB—10.5 3 mm 0 x l 5 m (l. 5 u m)

 Column temperature; initial temperature 230 ° C (0 min)

 Temperature rise + 2 ° C / min

 Final temperature 280 ° C (20 min)

 Inlet temperature; 300 ° C

 Detector temperature; 300 ° C

 Detector: Hydrogen ionization detector

 Split ratio; 1: 6

Machine used §5

 D e t e c t e r: Shimadzu G C-15 A (R an: 10)

 I n t e g r a t e r: S I C C h ro m a t o co r d e r 1 2

 (A t t. 3 2)

Reagents and reagents

 Test Stone Propionate (C22H32〇3)

 Made by SIGMA

 For determination / S—Sitosterol (C29H50O)

₉ Ma Biochemical Co., Ltd. (GLC purity 9 6.6%)

Quantitative results

 Table 1 shows the results of quantitative analysis of the lots for lots 1-7. [Table 1] Lot 3 ^ including β (%)

 1 0 .0 4 5 6

 2 0 .0 6 5 4

 3 0 .0 8 5 0

 4 0 .0 9 3 9

 5 0 .0 6 8 4

 6 0 .0 3 7 6

7 0 .0 4 6 1 Example 2

 In Example 1, 19 kg of the target extract was obtained in the same manner as in Example 1, except that 70% ethanol was replaced with 99% ethanol. Industrial applicability

 The extract of the present invention has a strong tonic and aphrodisiac effect, and a drug containing the extract of the present invention as an active ingredient is effective for the prevention or treatment of impotence and impaired energy due to stress. Test Example 1 (Quantification of fat-soluble components)

 [Sample]

 Specimen 1; 5 g of the kiss of Example 1

 Sample 2; 5 g of extract of Example 2

 Specimen 3; 5 g of Pequis obtained in the same manner as in Example 1 except that 70% ethanol was replaced with water.

 Specimen 4; 5 g of Pakis obtained in the same manner as in Example 1 except that 70% ethanol was replaced with 30% ethanol in Example 1.

 Specimen 5; 5 g of an extract obtained in the same manner as in Example 1 except that 70% ethanol was replaced with 50% ethanol in Example 1.

 Specimen 6; 5 g of commercially available dried kiss (manufactured by H. Finzelgerg Nachfogel Chemishe werke) Each of the specimens 1 to 5 other than the specimen 6 was preliminarily heated to 50 ° C. and subjected to a test as an almost uniform extract.

 The results of analyzing the main components of each sample are shown below. Determination of chemical substrate 台 by high-performance liquid chromatography

 Chemical compound A is determined in the example, though it is determined to be red. High-speed liquid chroma I is a special component in the graph method.

[Method for preparing sample solution] To each sample, add 5 ml of methanol, 5 ml of water, and 20 ml of n-hexane, shake for 10 minutes, centrifuge (300 rpm, 5 minutes), and remove the supernatant. The solution was concentrated under reduced pressure, and 25 ml of methanol was added to the obtained residue to prepare a sample solution. For quantification, ergosta-4, 6, 8 (14), 22-tetraen-3-one was used as a standard (3.5 mg was dissolved in 100 ml of methanol), It was quantified by the method of chromatography (HPLC).

 [High-speed liquid chromatograph conditions]

 Column; octadecylsilyl silicide gel gel (inner diameter 4 mm, length 150 mm)

 Detector; fluorescence detector

 Measurement wavelength; excitation wavelength 3 86 nm, fluorescence wavelength 4 72 nm

 Mobile phase; mixture of methanol and water (9: 1)

 Ram temperature; 50 ° C

 Injection volume: Quantitation of total steroids by 101 fluorometry

 [Method for preparing sample solution]

 To each sample, 5 ml of methanol, 5 ml of water and 20 ml of n-hexane are added, shaken for 10 minutes, centrifuged (30000 rpm, 5 minutes), and the supernatant is collected. After concentrating under reduced pressure, add 10 ml of a 1N hydroxylated water / water mixture (1: 1) to the resulting residue, add 20 ml X 3 of n-hexane, shake and add n-hexane. —-Heat cow liquid was collected and concentrated under reduced pressure, and 20 ml of sulfuric acid was added to the obtained residue to prepare a sample solution.

 The quantification is as follows:-As a standard product (3.5 mg dissolved in 100 ml of methanol), determine the fluorescence intensity (excitation wavelength: 480 nm, fluorescence intensity). 0 5 nm). Determination of 'class fatty acids by gas chromatography

 [Method for preparing sample solution]

To each sample, add 5 ml of methanol, 5 ml of water, and n- After shaking, centrifuge (300 rpm, 5 minutes), collect the supernatant, concentrate under reduced pressure 42563 1, and add 1N hydroxide Water mixture (1: 1) Add 10 ml of n-hexane, add 20 mlx3 of n-hexane, add 5 ml of hydrochloric acid and 20 m1 x 3 of n-hexane to the aqueous layer and shake. The n-hexane layer was removed, concentrated under reduced pressure, methylated with diazomethane monoether solution, the solvent was distilled off, and 2 ml of n-hexane was added to the obtained residue to prepare a sample solution. .

 Quantification was performed using stearic acid as a standard.

 [Gas chromatograph (GC) conditions]

 Column; methyl silicon polymer (inner diameter 0.53 mm, length 15 m) Detector; Hydrogen flame ionization detector

 Carrier gas; helium

 Flow rate: 20 m 1 Z min

 Power ram temperature; initial temperature 100 ° C, temperature rise 5 ° CZ, final temperature 300 ° C

 Injection volume; 1 1

 [Result]

 Table 2 shows the measurement results.

 [Table 2] Test items Compound A Total stearate higher fatty acids

 Test method HPLC method Fluorescence method G method

 0 3 2 0 7 2 0 0 9

 0 4 2 1 5 2 0 0 7

 0 0 0 0 0 0 3 0 0 2

 0 0 0 0 0 0 0 0 0 0

 0 0 6 0 0 7 0 0 6

 0 0 0 0 0 1 0 0 2 Test example 2

 [Sample]

 Specimen 7; 1 extract

Specimen 8; ΐ | ΐ 版 干 'i¾ extract (II. Finzel erg Nachfogoi Chemishc werkc: ¾!) [Test ίΜ] 4/02160

 -8-

One group of ICR mice (male, 8 weeks old) were used. ' [Test method]

 Rearing test male mice in isolation (4 weeks) causes sexual behavior disorders. After confirming the disorder 3, the subject was orally administered after 60 minutes, and a female mouse in estrus was placed 60 minutes later to observe sexual behavior.

 The test items are the following nine items.

 ①Mounting ②Mounting and ejaculation (intromission) ③Licking genitals (penislicking) す る Follow (（o1 1 owing) ⑤Grooming the other party (a11 1 gro om ing) ⑥Moving (1 ocomotion) ) ⑦Rising (rearing) ⑧Grooming yourself (se 1 f-grooming) 関心 Interest in genitals (anogenita 1)

 The above items were analyzed.

 [Result]

 ① Riding (ΙΏ 0 unting) ② Insertion and ejaculation (intromission) ③ About genital licking (penis 1 icking) Table 3 to Table 5 show the results of the number of animals that performed ① to ③ out of 8 animals per group. Show.

①Riding (mounting)

 [Table 3]

② '-In, ejaculation (intromission)

 [Table 4] Extract concentration% Sample 7 Sample 8

 01 2 5 1/8

 0.25 2/8 1/8

0.5 4/8 0/8 ③ licking genitals (penislicking)

 [Table 5]

Further, towards the specimen 1 also ④~⑨ showed a strong effect compared to sample 2 ₍

Claims

The scope of the claims . 1. a herb of Mui Lapuama  (A) Immersion liquid with 50% to 99% ethanol, and  (B) A solution obtained by soaking the residue of the immersion solution in 50% to 99% ethanol.  Concentrated — extract containing at least 0.03 3% sitosterol