CN101143185B - Medicinal composition capable of anti-senility during menopausal syndrome - Google Patents
Medicinal composition capable of anti-senility during menopausal syndrome Download PDFInfo
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- CN101143185B CN101143185B CN2007100292175A CN200710029217A CN101143185B CN 101143185 B CN101143185 B CN 101143185B CN 2007100292175 A CN2007100292175 A CN 2007100292175A CN 200710029217 A CN200710029217 A CN 200710029217A CN 101143185 B CN101143185 B CN 101143185B
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Abstract
The invention provides an antiaging drug combination for the menopause. The combination consists of active components and medically acceptable complementary materials, wherein the active components consist of the components with the following weight portions of 10 to 31 portions of epimedium herb extracts, 21 to 70 portions of yam extracts and 20 to 50 portions of soybean isoflavone extracts, wherein the epimedium herb extracts contain 20 to 90 percent of total flavones of the epimedium herb; the yam extracts contain 20 to 60 percent of diosgenin; the soybean isoflavone extracts contain 10 to 60 percent of soybean isoflavones. The combination of the invention can protect the women uterus and prevent the osteoporosis and effectively delay the menopause aging of women.
Description
Technical field
The present invention relates to a kind of medicinal preparation, being specifically related to Chinese medicine is the Chinese patent medicine of feedstock production.
Background technology
Climacteric syndrome is the women before and after menopause because the ovarian function decline, decrease in estrogen, promoting sexual gland hormone raises, and organism endocrine is readjusted in the process patient and can not be adapted to and cause the autonomic nervous dysfunction, with one group of syndrome of neural mental symptoms.Postmenopausal osteoporosis refers to that by the low osteoporosis that causes of postmenopausal estrogen level morbidity in 5～10 years after menopause often belongs to primary osteoporosis.It is characterized in that the minimizing of bone mineral content and matrix components equal proportion, the cortical bone attenuation, bone trabecula reduces, attenuates.The epidemiological study data shows: osteoporosis takes place in postmenopausal women about 30%.
At present the control medicine in climacteric syndrome has the estrogens medicine, but the type medicine is because of having the danger of diseases such as potential increase breast carcinoma, carcinoma of endometrium, and has limited its application.
In recent years, the treatment of climacteric syndrome that is found to be of phytoestrogen such as soybean isoflavone etc. provides new selection.Soybean isoflavone is to be present in the bioactive substance that a class in the Semen sojae atricolor has the effect of estrogen sample, comprises 12 kinds of chemical compounds altogether, accounts for 0.2%～0.4% of full Semen sojae atricolor.To the postmenopausal women because of the osteoporosis of serum estrogen level due to descending, early disease such as senile dementia and menopausal syndrome all has certain preventive and therapeutic effect.The soybean isoflavone treatment is still a kind of estrogenic exogenous replacement therapy that substitutes in essence, can correct to a certain extent because of estrogen the descend endocrine regulation cause, osteoporosis etc., but the mechanism that promotes estrogen production is not seen that the drug effect report is arranged.And many research report the kidney invigorating spleen reinforcing Chinese medicines can be adjusted hypothalamus-hypophysis-target gland s function by endogenous, and part reverses osteoporosis etc.
Summary of the invention
The technical problem to be solved in the present invention is the effect that further improves the anti-climacteric aging of soybean isoflavone.
In order to solve the problems of the technologies described above, the inventor filters out the Herba Epimedii with the kidney invigorating effect from numerous Chinese medicines, therefrom isolate the part that is rich in the Herba Epimedii total flavones component; Rhizoma Dioscoreae with spleen reinforcing effect is therefrom isolated the part that is rich in the diosgenin component; And above-mentioned two kinds of Chinese medicinal components and soybean isoflavone are made up in certain proportion, make the oral formulations of medically accepting.
According to above-mentioned design, the technical scheme that the present invention solves the problems of the technologies described above is:
A kind of climacteric antidotal pharmaceutical composition, said composition is made up of active component and medically acceptable adjuvant, and wherein said active component is made up of the component of following parts by weight: 10～31 parts of Herba Epimedii extracts, 21～70 parts of Rhizoma Dioscoreae extracts, 20～50 parts of soybean isoflavones;
Above-mentioned each component all is to be prepared by known method, and wherein Herba Epimedii extract is the pure molten or water-soluble part and refining the obtaining of extracting Herba Epimedii, contains 20～90% Herba Epimedii total flavones; Rhizoma Dioscoreae extract extracts petroleum ether dissolution and partly obtains behind acid hydrolysis, contain 20～60% diosgenins; Soybean isoflavone is that the pure molten part of extracting defatted soybean meal obtains, and contains 10～60% soybean isoflavone;
Above-mentioned percentage ratio all is weight percentage.
Preferred version of the present invention is: 20～31 parts of Herba Epimedii extracts, 31～70 parts of Rhizoma Dioscoreae extracts, 30～50 parts of soybean isoflavones; Contain Herba Epimedii total flavones 60～80% in the Herba Epimedii extract, contain diosgenin 20～60% in the Rhizoma Dioscoreae extract, contain soybean isoflavone 10～60% in the soybean isoflavone.
Preferred plan of the present invention is:
Pharmaceutical composition of the present invention, wherein active component is made up of following components in weight percentage: 31 parts of Herba Epimedii extracts, 70 parts of Rhizoma Dioscoreae extracts, 50 parts of soybean isoflavones; Contain Herba Epimedii total flavones 80% in the Herba Epimedii extract, contain diosgenin 20% in the Rhizoma Dioscoreae extract, contain soybean isoflavone 40% in the soybean isoflavone.
The preparation method of each component is as described below in the active ingredient in pharmaceutical of the present invention:
1, obtain after after Herba Epimedii extract can water decocts decocting liquid being purified by macroporous resin, concrete grammar is: take by weighing Herba Epimedii herb dry powder, add the water of 8～12 times of crude drug amounts, heat little boiling and extract 30～90min, cool off coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, filter.Filtrate is gone up macroporous resin column, with 2～5 times of volume distilled water washing resin posts, use the ethanol elution of 2～4 times of volumes 50～70% then, 50～70 ℃ are evaporated to driedly, get final product.
The Herba Epimedii extract of gained can also be used chloroform extraction (removing the little impurity of depolarization) and ethyl acetate extraction successively, gets to get yellow powder after ethyl acetate concentrates mutually, uses 60% ethyl alcohol recrystallization at last, can obtain the higher Herba Epimedii extract of purity.
2, Rhizoma Dioscoreae extract is earlier Rhizoma Dioscoreae to be obtained with Petroleum ether extraction after with acid hydrolysis, concrete grammar is: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, in crude drug: the ratio of 2.% sulphuric acid=1: 8～12, add isopyknic 2% sulphuric acid and petroleum ether, in 85～90 ℃ of hydrolysis 3～5h, incline to, cooling, layering, get upper strata (petroleum ether part), be washed till neutrality with sodium carbonate liquor, petroleum ether is reclaimed in reuse distillation washing, evaporate to dryness, reuse ethanol is washed, and reclaims ethanol, evaporate to dryness, put 50～70 ℃ of vacuum dryings, get final product;
Rhizoma Dioscoreae extract in the prescription of the present invention can also extract the Rhizoma Dioscoreae acid hydrolysis products and obtain by carbon dioxide supercritical extraction method.
3, the soybean isoflavone concrete grammar is: be raw material with the defatted soybean meal, pressing material ratio 1: 10～20 adds 60～80% ethanol, 60～80 ℃ of lixiviate 2～6h, filtering residue extracts 1 time again, merges 2 times filtrate, reclaims ethanol, by the macroporous resin adsorption eluting, by polyamide chromatography post, with 2 times of column volumes, 70% ethanol elution chromatography post, the eluent that receives is evaporated to dried getting final product under 65 ℃ behind the usefulness water wash pillar again.
Soybean isoflavone can also obtain by buying commercially available soybean isoflavone product.
Pharmaceutical composition of the present invention is common oral formulations, as soft capsule, drop pill etc.
Preparation of pharmaceutical compositions method of the present invention is: get Herba Epimedii extract, Rhizoma Dioscoreae extract and soybean isoflavone mix homogeneously by proportioning, add required adjuvant according to the requirement of the different preparations of preparation then, get final product.
Pharmaceutical composition of the present invention need carry out qualitative and detection by quantitative to the effective ingredient in the various raw extracts before preparation, concrete method is as follows:
(1) the qualitative and detection by quantitative of Herba Epimedii total flavones
With the icariin is reference substance, and with sample silica gel H thin layer chromatography, (10: 1: 1: 1) launch, the aluminum chloride qualitative reaction all showed the fluorescent red-orange speckle to ethyl acetate-butanone-formic acid-water, turns out to be Herba Epimedii total flavones.The employing of Herba Epimedii total flavones assay " carry out, and spectrophotography is measured trap at 270nm wavelength place, calculate general flavone content by the determination of total flavonoids method under Chinese pharmacopoeia version Herba Epimedii in 2005 item.
(2) the qualitative and detection by quantitative of diosgenin
Qualitative analysis
Adopt the method for thin layer chromatography.Developing solvent: V (petroleum ether): V (chloroform): V (methanol)=10: 10: 0.5.Expansion mode: ascending development, exhibition is apart from 8cm.Developer: 20g/L phosphomolybdic acid ethanol solution.Squirt profit, 140 ℃ of baking 5min are clear to the speckle colour developing.Diosgenin is effectively separated with other composition, is navy blue, and Rf value is about 0.32.
Quantitative analysis
The thin slice scan condition adopts single wavelength linear scanning, scanning wavelength 650nm on CAMAG Scanner 3; Slit 6.00 * 0.30mm; Scanning speed 20nm/s.
The mensuration of standard curve is carried out point sample with semi-automatic sample introduction instrument to the diosgenin reference substance solution, and the point sample amount is respectively 0.1,0.2,0.4,0.8,1.2,1.6,2.0,2.4,2.8 3.2 μ L put on the same block of efficient lamellae of silica gel G and detect, with diosgenin peak area (Y) is vertical coordinate, and sampling volume (V) is an abscissa, the drawing standard curve, regression equation is: Y=1.979V-229.474, r=0.9983; Good in .0.3～1.8 μ g scope internal linear relation.
Get 1g Rhizoma Dioscoreae extract 10ml dissolve with methanol, adopt the external standard two-point method that sample is measured; Write down the meansigma methods that 3 duplicate samples are measured for 3 times respectively.
(3) qualitative and quantitative analysis of soybean isoflavone
The qualitative analysis thin layer chromatography is used the silica GF254 lamellae, is developing solvent with chloroform-absolute methanol (10: 0.5), with standard substance and sample point sample (absolute methanol dissolving), detects under the 254nm uviol lamp, sees light yellow test point.
Standard curve is drawn: precision takes by weighing daidzein standard substance 2.0mg, places the 25mL volumetric flask, with dissolve with methanol and be settled to groove, shakes up.The accurate respectively standard solution 0.1,0.2,0.3,0.4,0.6 of drawing, 0.8mL places the 10ml volumetric flask, is diluted to groove with methanol, shakes up.With methanol is blank, measures the A value at the wavelength place of 254nm.With concentration is abscissa, and the A value is a vertical coordinate drawing standard curve, gets regression equation Y=0.138X-0.0056, R=0.9994, the range of linearity 0.8～6.4mg/L.
Quantitative analysis: precision takes by weighing the 0.5g soybean isoflavone, puts in the 100mL round-bottomed flask, adds 80% methanol 70ml supersound extraction 40min, sucking filtration while hot, methanol extract liquid then at 45 ℃ of pressure reducing and steaming water, adds the 20mL dissolve with methanol as test sample at 35 ℃ of pressure reducing and steaming methanol again.Accurately measure need testing solution 0.01ml, arrive the 10ml standardize solution for 1000 times with the absolute methanol dilution, measure A value 3 times at wavelength 254nm place with ultraviolet spectrophotometer, average, measurement result is according to standard curve calculation sample content.
The same available said method of the active component of pharmaceutical composition finished product of the present invention is identified.
Chinese medicine thinks that the Woman climacteric aging mainly belongs to the depletion of the congenital foundation kidney essense gas and the void thanks to of the foundation of acquired constitution temper.The present composition is an active component with soybean isoflavone, Herba Epimedii extract and Rhizoma Dioscoreae extract, wherein mainly contain soybean isoflavone in the soybean isoflavone, soybean isoflavone has weak estrogen activity, can combine with estrogen receptor, when human body inner estrogen level is low, soybean isoflavone occupies estrogen receptor, and the weak estrogen effect of performance shows the effect that improves estrogen level.Soybean isoflavone can combine with the estrogen receptor on the osteoclast, reduces its activity, and bone-loss is reduced.Mainly contain Herba Epimedii total flavones in the Herba Epimedii extract, Herba Epimedii total flavones then can be adjusted the hypothalamus monoamine transmitters, promote functions such as adenohypophysis, gonad, thereby regulate the hypothalamic pituitary gonadal axis function in all directions; The basic structural similarity of steroid hormone molecule in diosgenin that is rich in the Rhizoma Dioscoreae extract and the body after being absorbed by the body, can promote to comprise the formation of the biosynthetic precursors of steroid hormone such as gonadal hormone, 17-hydroxy-11-dehydrocorticosterone.The present composition is in soybean isoflavone adjusting self estrogen secretion, utilize the effect of the flat spleen reinforcing kidney of merit, Rhizoma Dioscoreae of Herba Epimedii kidney invigorating and YANG supporting, wind-damp dispelling, bone and muscle strengthening to treat the depletion of this kidney essense gas and the void thanks to of the foundation of acquired constitution temper, fundamentally prevent and treat climacteric syndrome, reach the purpose of delaying female climacteric aging.
Experimental results show that, the metratrophia pathological changes of the effective antagonism removal ovary of pharmaceutical composition of the present invention rat, the estrogen of rising removal ovary rat, delay the removal ovary rat the osteoporosis pathological changes, delay the old and feeble of removal ovary rat uterus and delay the adenohypophyseal aging of removal ovary rat, its effect is all effective than simple use soybean isoflavone.As seen, the present invention can protect the women uterus, prevent osteoporosis, effectively delaying female climacteric aging.
To prove the technique effect that the present invention has by zoopery below.
Pharmacodynamic experiment
(1) materials and methods
1 material
Laboratory animal: 3 month female SD rats, body weight 300g～320g.
Medicine for treatment:
Soft capsule of the present invention, every soft capsule nearly weighs the 500mg/ grain, every contains 60mg soybean isoflavone, 60mg Herba Epimedii extract, 140mg Rhizoma Dioscoreae extract, wherein soybean isoflavone contains soybean isoflavone 40%, Herba Epimedii extract contains Herba Epimedii total flavones 70%, and Rhizoma Dioscoreae extract contains diosgenin 20%.
2 experimental techniques
2.1 the animal grouping: rat is divided into 4 groups by body weight:
A organizes (normal group): the normal female rat, and 10, irritate stomach once with Oleum Glycines every day, continued for 12 weeks;
B organizes (removal ovary model group): removal ovary female rats, 10;
C group (simple soybean isoflavone treatment group): the removal ovary female rats, 10, give the 5mg soybean isoflavone every day and irritate the stomach treatment, continued for 12 weeks, soybean isoflavone is with Oleum Glycines dissolving and dilution before the experiment, and dosage is equivalent to 10 times of clinical adult (60kg) consumption;
D organizes (soft capsule treatment group of the present invention), the removal ovary female rats, and 10, give 50mg soft capsule of the present invention (including 5mg soybean isoflavone, 5mg Herba Epimedii total flavones extract, 10mg diosgenin extract) every day and irritate the stomach treatment, continued for 12 weeks; Soft capsule of the present invention dissolves and dilution with Oleum Glycines before experiment.
2.2 removal ovary modeling method: lumbar injection 1% pentobarbital sodium anesthesia, female rats, get under the most last rib after the anesthesia, midaxillary line and apart from the about 2cm infall in the spinal column outside, wipe out and become mildewed, cut skin and the about 1.5～2cm of dorsal muscles, with fallopian tube under the ovary (comprising fat) silk thread ligation, extract ovary.Postoperative is with iodine tincture wiping wound, intramuscular injection one all penicillins.The continuous 3d of postoperative makes vaginal smear examination, comes off epithelial cell as the index of oophorectomize success not see maturation in the smear, is used for subsequent experimental.
2.3 blood E2, FSH, LH detect: rat moment broken end is got blood, leaves standstill 4h, centrifugalize serum ,-20 ℃ of preservations.
2.4 uterus index: behind the sacrifice of animal, cut off stomach wall, expose the abdominal cavity, whether complete, and complete separating uterus if observing the animal ovary excision, cut off in uterus and vagina intersection.Remove fat and connective tissue around the uterus, after the normal saline washing, filter paper blots the uterus surface moisture.Use 1/10000 scales/electronic balance weighing.Uterus index=uterus weight in wet base/body weight.
2.5 rat uterus pathological examination: take out rat uterus; Behind the above-mentioned tissue physiology salt water rinse, filter paper blots surface moisture, places neutral formalin solution to fix by group.Paraffin embedding, serial section is all chosen middle part, uterus, both sides palace soma when draw materials in the uterus, slice thickness 4 μ m, HE dyeing, 10 * 10 light microscopics are observed the rat endometrium situation of change down.
2.6 rat bone density detects: get the bilateral femur, thoroughly reject soft tissue.With the U.S. QDR2000 of HOLOGIG company type dual intensity X line absorption instrument, measure the bilateral femoral bmd.
2.7 statistical method: each is organized detected value and represents with mean and standard deviation, relatively adopt one factor analysis of variance between each group of homogeneity of variance parameter, for comparing between the group of heterogeneity of variance parameter, adopt the Kruskal-Wallis H check in a plurality of independent sample non parametric testss.
(2) result
1 respectively organizes rat body weight, heavy, the uterus index variation in uterus, sees Table 1.
Table 1 is respectively organized rat body weight, the uterus heavily changes.
Compare #P＜0.05, ##P＜0.01 with normal group; Compare with the removal ovary model group, ▲ P＜0.05, ▲ ▲ P＜0.01.
Female rats after spay, body weight than the same period normal group obviously increase (p＜0.01), and uterus weight, uterus coefficient obviously alleviate (p＜0.01); Soybean isoflavone treatment group, soft capsule treatment group body weight of the present invention have been compared with the removal ovary model group and have been alleviated trend, but not statistically significant; Soybean isoflavone treatment group, soft capsule treatment group of the present invention uterus weight and uterus coefficient are apparently higher than model group (p＜0.05).
This results suggest, soft capsule treatment of the present invention is the metratrophia pathological changes of antagonism removal ovary rat effectively, and simple soybean isoflavone treatment curative effect obviously strengthens.
2 respectively organize rat E2, FSH, LH variation, see Table 2.
Table 2 is respectively organized rat E2, FSH, LH changing value.
Compare #P＜0.05, ##P＜0.01 with normal group; Compare with the removal ovary model group, ▲ P＜0.05, ▲ ▲ P＜0.01.
Compare with normal group, E2 obviously reduces (p＜0.01) in the ovariectomized group serum, and FSH raises, and LH obviously reduces (p＜0.01); Soybean isoflavone treatment group E2 obviously raises (p＜0.05) than ovariectomized group, and FSH reduces (p＜0.05), LH slightly raises; E2 concentration is higher than simple soybean isoflavone treatment group in the soft capsule treatment group serum of the present invention, and FSH is lower than simple soybean isoflavone treatment group, LH is higher than simple soybean isoflavone treatment group.
This results suggest, soft capsule of the present invention is treated the estrogen of the removal ovary rat that can effectively raise, and simple soybean isoflavone treatment curative effect obviously strengthens.
Table 3 is respectively organized rat femur bone density (g/cm
2) changing value.
Compare #P＜0.05, ##P＜0.01 with normal group; Compare with the removal ovary model group, ▲ P＜0.05, ▲ ▲ P＜0.01.
Compare with normal group, ovariectomized group rat femur bone density obviously reduces (p＜0.01); Soybean isoflavone treatment group is than ovariectomized group obviously raise (p＜0.05); Soft capsule treatment group bone density of the present invention is better than simple soybean isoflavone treatment group.
This results suggest, soft capsule treatment of the present invention can effectively delay the osteoporosis pathological changes of removal ovary rat, and simple soybean isoflavone treatment curative effect strengthens.
4 respectively organize the rat uterus pathologic finding
4.1 rat uterus pathological examination (10 * 10)
Normal rat uterus body wall is divided into theca interna, flesh layer and placenta percreta from inside to outside.
The normal rats endometrium is made of the high columnar epithelial cell of monolayer, and intimal epithelium does not have atrophy or hypertrophy, is distributed with the simple tubular gland body in the inner membrance lamina propria, and glandular epithelium is the monolayer column, and nucleus is rounded, is positioned at the cell based bottom; Matter inner cell quantity is many between lamina propria, and a matter is loose, as shown in Figure 1.
Ovariectomized group rat endometrium and flesh layer be significantly attenuation all, the obvious atrophy of theca interna, and intimal epithelium is made of flat or cuboid cell, and part intimal epithelium atrophy is extremely not obvious; Body of gland quantity obviously reduces in the inner membrance lamina propria, and the body of gland chamber obviously reduces, and is bordering on closure, and glandular epithelium is obviously atrophy also, and the Interstitial cell volume reduces, and a matter is tight relatively, as shown in Figure 2.
Soybean isoflavone treatment group endometrium treatment group endometrial epithelium, glandular epithelium, Interstitial cell all have the atrophy pathological changes, but obviously are lighter than ovariectomized group, still as seen have the simple tubular gland body to distribute at lamina propria, as shown in Figure 3.
Soft capsule treatment group endometrial epithelium of the present invention, glandular epithelium, above each group of Interstitial cell atrophy pathological changes obviously alleviate, and as seen have the simple tubular gland body to distribute at lamina propria, and a matter is also more loose, as shown in Figure 4.
The above results prompting soft capsule treatment of the present invention drug effect is obvious, and simple soybean isoflavone treatment curative effect strengthens, and can effectively delay the aging of removal ovary rat uterus.
4.2 (10 * 10) are observed in rat adenohypophysis pathology HE dyeing
Adenohypophysis divides oxyphil cell, basophil, chromophobe cell three kinds, and wherein the oxyphil cell is divided into somatotroph and mammotroph according to the reaction of its granular size and SABC; Basophil is divided into thyrotroph, TSH cell, gonadotroph, corticotroph, ACTH cell; Chromophobe cell is for stocking sexual cell or secretion back cell.
The normal group adenohypophysis is made up of a large amount of glandular cells, and cell is a strand and distributes, and sinusoid capillary is arranged between the cell mass.What cell quantity was maximum is that cell space is less, the painted more shallow chromophobe cell of kytoplasm; Cell space is bigger, the kytoplasm red colouration be the oxyphil cell; Cell space is bigger, and kytoplasm dyes the indigo basophil that is, as shown in Figure 5.
The obvious atrophy of all glandular cell cell spaces of ovariectomized group rat adenohypophysis, blood capillary hole distribute and also reduce, as shown in Figure 6.
Soybean isoflavone treatment group, soft capsule treatment group adenohypophysis of the present invention do not have obvious atrophy phenomenon, point out above-mentioned treatment can effectively delay the adenohypophyseal aging of removal ovary rat, shown in Fig. 7,8.
In sum, the metratrophia pathological changes of the effective antagonism removal ovary of pharmaceutical composition of the present invention rat, the estrogen of rising removal ovary rat, delay the removal ovary rat the osteoporosis pathological changes, delay the old and feeble of removal ovary rat uterus and delay the adenohypophyseal aging of removal ovary rat, its effect is all effective than simple use soybean isoflavone.As seen, the present invention can protect the women uterus, prevent osteoporosis, effectively delaying female climacteric aging.
Description of drawings
Fig. 1 is a normal rats uterus pathological examination microgram (10 * 10);
Fig. 2 is an ovariectomized group rat uterus pathological examination microgram (10 * 10);
Fig. 3 is a soybean isoflavone treatment removal ovary rat uterus pathological examination microgram (10 * 10);
Fig. 4 is a soft capsule treatment removal ovary rat uterus pathological examination microgram (10 * 10) of the present invention;
Fig. 5 is a normal rats adenohypophysis pathology HE colored graph;
Fig. 6 is a removal ovary rat adenohypophysis pathology HE colored graph;
Fig. 7 is a soybean isoflavone treatment removal ovary rat adenohypophysis pathology HE colored graph;
Fig. 8 is a soft capsule treatment removal ovary rat adenohypophysis pathology HE colored graph of the present invention.
The specific embodiment
Example 1
1, the preparation of each component in the active component:
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, add the distilled water of 10 times of amounts, heat little boiling and extract 30min, cooling, coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, sucking filtration.The pretreated D101 macroporous adsorbent resin of learning from else's experience, wet method dress post is washed standby only.With D101 resin column on the Herba Epimedii water extract,, use the ethanol elution of 3 times of volumes 50% then with 5 times of volume distilled water washing resin posts.Will the solution behind resin elution use chloroform extraction successively 3 times (removing the little impurity of depolarization) and ethyl acetate extraction 3 times, the consumption of each extractant are to treat 2 times of extract volume, and the combined ethyl acetate phase must yellow powder after concentrating.With above-mentioned yellow powder 60% ethyl alcohol recrystallization, obtain the faint yellow extract that contains 90% Herba Epimedii total flavones.
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of 2% sulphuric acid=1: 8 adds isopyknic 2% sulphuric acid, in 85 ℃ of hydrolysis 3h, be washed till neutrality with sodium carbonate liquor, petroleum ether is reclaimed in reuse distillation washing, evaporate to dryness moves into extractor, with CO
2Supercritical extraction extracts.With pressure 35.0MPa CO
2Gas enters CO through filter
2Cooling liquid in the condenser pumps into pipeline then; Simultaneously with volume fraction be 95% ethanol for carrying agent, carry agent content 3%, carry hydrolyzate to be extracted and pump into pipeline by another; CO
2With carry agent and enter extractor through threeway, leave standstill at 45 ℃ of constant temperature and begin Dynamic Separation after soaking 6h, contain the CO of extract
2Blood pressure lowering enters separator to gas through needle-valve, at 60 ℃ of following Dynamic Separation 3h of normal pressure, CO
2Flow velocity 2Lmin
-1, CO
2Through discharging by air vent behind the volume metering meter, separate obtaining Rhizoma Dioscoreae extract, wherein contain diosgenin 60%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, press material ratio and add 60～80% ethanol at 1: 10,60 ℃ of lixiviate 2h, filtering residue extracts 1 time again, merge 2 times filtrate, reclaim ethanol, use the AB-8 adsorption and desorption by resin, AB-8 resin column on the solution, volume 40% alcoholic solution with 3 times of resins carries out eluting, collects eluent, and the soybean isoflavone solution behind the recovery ethanol is by polyamide chromatography post, use the water wash pillar, with 2 times of column volumes, 70% ethanol elution chromatography post, the eluent that receives is evaporated to do under 65 ℃ promptly obtains isoflavone extract, wherein contain soybean isoflavone 60%.
2, preparation soft capsule:
(1) get Herba Epimedii extract 110g, Rhizoma Dioscoreae extract 700g, soybean isoflavone 200g with the mixing of 1500g Oleum Glycines, it is even to stir mill.Get the edible Cera Flava (as emulsifying agent) of 200g and add Oleum Glycines heating fusion, above-mentioned two parts are stirred.
(2) preparation pharmagel: the preheating glue pot, add pure water 30kg, glycerol 13.5kg, be heated to 82 ℃, add gelatin 30kg and ethylparaben 0.15kg.Stir and evacuation, continue evacuation behind the liquid level stabilizing, vacuum is greater than 0.07mpa.Sampling observation glue quality, it is standby to put glue after qualified.Take by weighing the iron oxide red glycerol adding, fuse overgroundly, pour in the gelatin and mix, stir.
(3) carry out pelleting with the pelleting method, rubber thickness 80mm, 60 ℃ of glue temperatures, 44 ℃ of sprinkler body temperature.
(4) the dried soft gelatin capsule of will finalizing the design is washed ball with ethanol in soft capsule washing machine.
(5) above-mentioned raw materials can make 6600 altogether, and the soft capsule of making nearly weighs the 500mg/ grain, and every contains the 200mg ingredient, takes 1～2 every day.
Example 2
1, the preparation of each component in the active component:
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, use the alcohol reflux of 10 times of volumes 70% then, the collection backflow filters and also reclaims ethanol, and thin film rotary evaporator concentrates and the dry Herba Epimedii extract that gets, and wherein contains Herba Epimedii total flavones 20%.
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of 2% sulphuric acid=1: 12 adds isopyknic 2% sulphuric acid and petroleum ether, in 90 ℃ of hydrolysis 5h, incline to, cooling, layering, get upper strata (petroleum ether part), be washed till neutrality with sodium carbonate liquor, reuse distillation washing is reclaimed petroleum ether, evaporate to dryness, reuse ethanol is washed, reclaim ethanol, evaporate to dryness is put 70 ℃ of vacuum dryings, get Rhizoma Dioscoreae extract, wherein contain diosgenin 20%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, presses material ratio and adds 60% ethanol, lixiviate 6h at 1: 18,70 ℃ of extraction temperatures, filtering residue extract 1 time again, merge 2 times filtrate, reclaim ethanol, get soybean isoflavone, wherein contain soybean isoflavone 10%.
2, preparation soft capsule:
(1) get Herba Epimedii extract 310g, Rhizoma Dioscoreae extract 500g, soybean isoflavone 500g with the mixing of 2000g Oleum Glycines, it is even to stir mill.Get the edible Cera Flava (as emulsifying agent) of 300g and add Oleum Glycines heating fusion, above-mentioned two parts are stirred.
(2) preparation pharmagel: the preheating glue pot, add pure water 30kg, glycerol 13.5kg, be heated to 82 ℃, add gelatin 30kg and ethylparaben 0.15kg.Stir and evacuation, continue evacuation behind the liquid level stabilizing, vacuum is greater than 0.07mpa.Sampling observation glue quality, it is standby to put glue after qualified.Take by weighing the iron oxide red glycerol adding, fuse overgroundly, pour in the gelatin and mix, stir.
(3) carry out pelleting with the pelleting method, rubber thickness 80mm, 60 ℃ of glue temperatures, 44 ℃ of sprinkler body temperature.
(4) the dried soft gelatin capsule of will finalizing the design is washed ball with ethanol in soft capsule washing machine.
(5) soft capsule made of above-mentioned raw materials nearly weighs the 500mg/ grain, and every contains the 200mg ingredient, takes 2～4 every day.
Example 3
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, add the distilled water of 10 times of amounts, heat little boiling and extract 30min, cooling, coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, sucking filtration.With D101 resin column on the Herba Epimedii water extract (resin heavy 100g), with 5 times of volume distilled water washing resin posts, use the ethanol elution of 3 times of volumes 50% then, 60 ℃ are evaporated to driedly, and Herba Epimedii extract wherein contains Herba Epimedii total flavones 70%
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of hydrolyzed solution=1: 10 adds isopyknic 2% sulphuric acid and petroleum ether, in 88 ℃ of hydrolysis 4h, incline to, cooling, layering, get upper strata (petroleum ether part), be washed till neutrality with sodium carbonate liquor, reuse distillation washing is reclaimed petroleum ether, evaporate to dryness, reuse ethanol is washed, reclaim ethanol, evaporate to dryness is put 60 ℃ of vacuum dryings, get Rhizoma Dioscoreae extract, wherein contain diosgenin 20%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, presses material ratio and adds 60% ethanol, 70 ℃ of lixiviate 4h at 1: 15,, filtering residue extracts 1 time again, merges 2 times filtrate, reclaim ethanol, AB-8 resin column on the solution, last column volume is equivalent to the volume of resin in the post, carries out eluting with volume 40% alcoholic solution of 3 times of resins, collect eluent, 60 ℃ be evaporated to dried, soybean isoflavone, wherein contain soybean isoflavone 40%.
2, preparation soft capsule:
(1) get Herba Epimedii extract 310g, Rhizoma Dioscoreae extract 700g, soybean isoflavone 500g with the mixing of 2500g Oleum Glycines, it is even to stir mill.Get the edible Cera Flava (as emulsifying agent) of 300g and add Oleum Glycines heating fusion, above-mentioned two parts are stirred.
(2) preparation pharmagel: the preheating glue pot, add pure water 30kg, glycerol 13.5kg, be heated to 82 ℃, add gelatin 30kg and ethylparaben 0.15kg.Stir and evacuation, continue evacuation behind the liquid level stabilizing, vacuum is greater than 0.07mpa.Sampling observation glue quality, it is standby to put glue after qualified.Take by weighing the iron oxide red glycerol adding, fuse overgroundly, pour in the gelatin and mix, stir.
(3) carry out pelleting with the pelleting method, rubber thickness 80mm, 60 ℃ of glue temperatures, 44 ℃ of sprinkler body temperature.
(4) the dried soft gelatin capsule of will finalizing the design is washed ball with ethanol in soft capsule washing machine.
(5) soft capsule made of above-mentioned raw materials nearly weighs the 500mg/ grain, and every contains the 200mg ingredient, takes 1～2 every day.
Example 4
1, the preparation of each component in the active component:
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, add the distilled water of 10 times of amounts, heat little boiling and extract 30min, cooling, coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, sucking filtration.Adopt the AB-8 macroporous adsorbent resin, adopting 70% ethanol during eluting is eluant, and the eluant consumption is 6 times of amount resin volumes, eluting, and 60 ℃ are evaporated to driedly, and Herba Epimedii extract wherein contains Herba Epimedii total flavones 80%.
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of hydrolyzed solution=1: 9 adds isopyknic 2% sulphuric acid, in 85 ℃ of hydrolysis 6h, be washed till neutrality with sodium carbonate liquor, petroleum ether is reclaimed in reuse distillation washing, evaporate to dryness moves into extractor, with CO
2Supercritical extraction extracts.With pressure 35.0MPa CO
2Gas enters cooling liquid in the CO2 condenser through filter, pumps into pipeline then; Simultaneously with volume fraction be 95% ethanol for carrying agent, carry agent content 3%, carry hydrolyzate to be extracted and pump into pipeline by another; CO2 and carry agent and enter extractor through threeway, after leaving standstill immersion 6h, 45 ℃ of constant temperature begin Dynamic Separation, the CO2 gas process needle-valve blood pressure lowering that contains extract enters separator, Dynamic Separation 3h under normal pressure 60C, CO2 flow velocity 2Lmin-1, discharge by air vent behind the CO2 process volume metering meter, separate Rhizoma Dioscoreae extract, wherein contain diosgenin 60%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, press material ratio and add 80% ethanol at 1: 18,65 ℃ of lixiviate 5h, filtering residue extracts 1 time again, merges 2 times filtrate, reclaims ethanol, use the AB-8 adsorption and desorption by resin, AB-8 resin column on the solution, last column volume is equivalent to the volume of resin in the post, carries out eluting with volume 40% alcoholic solution of 3 times of resins, collect eluent, soybean isoflavone solution behind the recovery ethanol is used the water wash pillar, with 2 times of column volumes, 70% ethanol elution chromatography post by polyamide chromatography post, the eluent that receives is evaporated to do under 65 ℃ promptly obtains isoflavone extract, wherein contain soybean isoflavone 60%.
2, preparation drop pill:
Get Herba Epimedii extract 100g, Rhizoma Dioscoreae extract 210g, soybean isoflavone 300g.In Polyethylene Glycol and 3: 1 ratio of drug ratios, the 6000g polyethylene glycol 6000 is heated to 90 ℃, after treating whole fusions, add above-mentioned three kinds of Chinese medicine extract and be stirred to fusion, be transferred in the reservoir, airtight and the insulation at 90 ℃, regulate the dropping liquid quantitative valve, from top to bottom, splash in 10 ℃～15 ℃ the liquid Paraffin, with the drop pill drop that is shaped to the greatest extent and wipe liquid Paraffin, dry and storage.Manufactured goods are yellow drop pill, specification: every 50mg, effective ingredient 12.5mg/ grain is taken 3 every day, takes 6 at every turn.
Example 4
1, the preparation of each component in the active component:
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, add the distilled water of 10 times of amounts, heat little boiling and extract 30min, cooling, coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, sucking filtration.Adopt the AB-8 macroporous adsorbent resin, adopting 70% ethanol during eluting is eluant, and the eluant consumption is 6 times of amount resin volumes, eluting, and 60 ℃ are evaporated to driedly, and Herba Epimedii extract wherein contains Herba Epimedii total flavones 80%.
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of hydrolyzed solution=1: 9 adds isopyknic 2% sulphuric acid, in 85 ℃ of hydrolysis 6h, be washed till neutrality with sodium carbonate liquor, petroleum ether is reclaimed in reuse distillation washing, evaporate to dryness moves into extractor, with CO
2Supercritical extraction extracts.With pressure 35.0MPa CO
2Gas enters cooling liquid in the CO2 condenser through filter, pumps into pipeline then; Simultaneously with volume fraction be 95% ethanol for carrying agent, carry agent content 3%, carry hydrolyzate to be extracted and pump into pipeline by another; CO2 and carry agent and enter extractor through threeway, after leaving standstill immersion 6h, 45 ℃ of constant temperature begin Dynamic Separation, the CO2 gas process needle-valve blood pressure lowering that contains extract enters separator, at 60 ℃ of following Dynamic Separation 3h of normal pressure, CO2 flow velocity 2Lmin-1, discharge by air vent behind the CO2 process volume metering meter, separate Rhizoma Dioscoreae extract, wherein contain diosgenin 60%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, press material ratio and add 80% ethanol at 1: 18,65 ℃ of lixiviate 5h, filtering residue extracts 1 time again, merges 2 times filtrate, reclaims ethanol, use the AB-8 adsorption and desorption by resin, AB-8 resin column on the solution, last column volume is equivalent to the volume of resin in the post, carries out eluting with volume 40% alcoholic solution of 3 times of resins, collect eluent, soybean isoflavone solution behind the recovery ethanol is used the water wash pillar, with 2 times of column volumes, 70% ethanol elution chromatography post by polyamide chromatography post, the eluent that receives is evaporated to do under 65 ℃ promptly obtains isoflavone extract, wherein contain soybean isoflavone 60%.
2, preparation drop pill:
Get Herba Epimedii extract 300g, Rhizoma Dioscoreae extract 210g, soybean isoflavone 200g.In Polyethylene Glycol and 3: 1 ratio of drug ratios, the 4500g polyethylene glycol 6000 is heated to 100 ℃, after treating whole fusions, add above-mentioned three kinds of Chinese medicine extract and be stirred to fusion, be transferred in the reservoir, airtight and the insulation at 90 ℃, regulate the dropping liquid quantitative valve, from top to bottom, splash in 10 ℃～15 ℃ the liquid Paraffin, with the drop pill drop that is shaped to the greatest extent and wipe liquid Paraffin, dry and storage.Manufactured goods are yellow drop pill, specification: every 50mg, effective ingredient 12.5mg/ grain is taken 3 every day, takes 6 at every turn.
Example 6
1, the preparation of each component in the active component:
(1) Herba Epimedii extract: take by weighing Herba Epimedii herb dry powder, add the distilled water of 10 times of amounts, heat little boiling and extract 30min, cooling, coarse filtration; Filtering residue repeats to extract 1 time; Merge coarse filtration liquid twice, sucking filtration.Adopt the AB-8 macroporous adsorbent resin, adopting 70% ethanol during eluting is eluant, and the eluant consumption is 6 times of amount resin volumes, eluting, and 60 ℃ are evaporated to driedly, and Herba Epimedii extract wherein contains Herba Epimedii total flavones 80%.
(2) Rhizoma Dioscoreae extract: the Rhizoma Dioscoreae decoction pieces is crushed to about 10 orders, and in crude drug: the ratio of hydrolyzed solution=1: 9 adds isopyknic 2% sulphuric acid, in 85 ℃ of hydrolysis 6h, be washed till neutrality with sodium carbonate liquor, petroleum ether is reclaimed in reuse distillation washing, evaporate to dryness moves into extractor, with CO
2Supercritical extraction extracts.With pressure 35.0MPa CO
2Gas enters cooling liquid in the CO2 condenser through filter, pumps into pipeline then; Simultaneously with volume fraction be 95% ethanol for carrying agent, carry agent content 3%, carry hydrolyzate to be extracted and pump into pipeline by another; CO2 and carry agent and enter extractor through threeway, after leaving standstill immersion 6h, 45 ℃ of constant temperature begin Dynamic Separation, the CO2 gas process needle-valve blood pressure lowering that contains extract enters separator, at 60 ℃ of following Dynamic Separation 3h of normal pressure, CO2 flow velocity 2Lmin-1, discharge by air vent behind the CO2 process volume metering meter, separate Rhizoma Dioscoreae extract, wherein contain diosgenin 60%.
(3) soybean isoflavone: with the defatted soybean meal is raw material, press material ratio and add 80% ethanol at 1: 18,65 ℃ of lixiviate 5h, filtering residue extracts 1 time again, merges 2 times filtrate, reclaims ethanol, use the AB-8 adsorption and desorption by resin, AB-8 resin column on the solution, last column volume is equivalent to the volume of resin in the post, carries out eluting with volume 40% alcoholic solution of 3 times of resins, collect eluent, soybean isoflavone solution behind the recovery ethanol is used the water wash pillar, with 2 times of column volumes, 70% ethanol elution chromatography post by polyamide chromatography post, the eluent that receives is evaporated to do under 65 ℃ promptly obtains isoflavone extract, wherein contain soybean isoflavone 60%.
2, preparation drop pill:
Get Herba Epimedii extract 210g, Rhizoma Dioscoreae extract 700g, soybean isoflavone 300g.In Polyethylene Glycol and 3: 1 ratio of drug ratios, the 4200g polyethylene glycol 6000 is heated to 90 ℃, after treating whole fusions, add above-mentioned three kinds of Chinese medicine extract and be stirred to fusion, be transferred in the reservoir, airtight and the insulation at 90 ℃, regulate the dropping liquid quantitative valve, from top to bottom, splash in 10 ℃～15 ℃ the liquid Paraffin, with the drop pill drop that is shaped to the greatest extent and wipe liquid Paraffin, dry and storage.Manufactured goods are yellow drop pill, specification: every 50mg, effective ingredient 12.5mg/ grain is taken 3 every day, takes 6 at every turn.
Claims (4)
- One kind climacteric antidotal pharmaceutical composition, said composition is made up of active component and medically acceptable adjuvant, and wherein said active component is made up of the component of following parts by weight: 10～31 parts of Herba Epimedii extracts, 21～70 parts of Rhizoma Dioscoreae extracts, 20～50 parts of soybean isoflavones; Wherein Herba Epimedii extract is the pure molten or water-soluble part and refining the obtaining of extracting Herba Epimedii, contains 20～90% Herba Epimedii total flavones; Rhizoma Dioscoreae extract extracts petroleum ether dissolution with Rhizoma Dioscoreae and partly obtains behind acid hydrolysis, contain 20～60% diosgenins; Soybean isoflavone is that the pure molten part of extracting defatted soybean meal obtains, and contains 10～60% soybean isoflavone;Above-mentioned percentage ratio all is weight percentage.
- 2. a kind of climacteric according to claim 1 antidotal pharmaceutical composition, it is characterized in that described active component is obtained by following feedstock production: 20～31 parts of Herba Epimedii extracts, 31～70 parts of Rhizoma Dioscoreae extracts, 30～50 parts of soybean isoflavones; Wherein contain Herba Epimedii total flavones 20～90% in the Herba Epimedii extract, contain diosgenin 20～60% in the Rhizoma Dioscoreae extract, contain soybean isoflavone 10～60% in the soybean isoflavone.
- 3. a kind of climacteric according to claim 1 antidotal pharmaceutical composition, it is characterized in that described active component is obtained by following feedstock production: 31 parts of Herba Epimedii extracts, 70 parts of Rhizoma Dioscoreae extracts, 50 parts of soybean isoflavones; Wherein contain Herba Epimedii total flavones 80% in the Herba Epimedii extract, contain diosgenin 20% in the Rhizoma Dioscoreae extract, contain soybean isoflavone 40% in the soybean isoflavone.
- 4. according to antidotal pharmaceutical composition of claim 1,2 or 3 described a kind of climacteric, said composition is soft capsule or drop pill.
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| CN103566129A (en) * | 2012-11-10 | 2014-02-12 | 武小力 | Chinese and Western combined prescription for improving female climacteric syndrome and preparation method thereof |
| CN105879014A (en) * | 2014-10-23 | 2016-08-24 | 大连世纪新源技术开发有限公司 | Deuterium-depleted water (DDW) oral liquid with functions of invigorating kidney and preventing aging |
| CN109350624A (en) * | 2018-05-25 | 2019-02-19 | 泓博元生命科技（深圳）有限公司 | Female health composition, preparation containing NMN and DHEA and the preparation method and application thereof |
| CN109364171A (en) * | 2018-05-25 | 2019-02-22 | 泓博元生命科技（深圳）有限公司 | Ovary maintaining composition, preparation containing NADH and DHEA and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1072855A (en) * | 1992-12-02 | 1993-06-09 | 杨晶珠 | Prevent and treat climacteric syndrome medicaments preparation method |
| CN1038813C (en) * | 1993-07-02 | 1998-06-24 | 周勇 | Traditional Chinese medicine for curing osteoporosis |
| CN1368145A (en) * | 2001-02-02 | 2002-09-11 | 杨孟君 | Nano medicine 'Gengnianshu' and its preparing process |
| CN1569017A (en) * | 2003-07-24 | 2005-01-26 | 复旦大学附属华山医院 | Total epimeddium flavone tablet and its preparation method |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1072855A (en) * | 1992-12-02 | 1993-06-09 | 杨晶珠 | Prevent and treat climacteric syndrome medicaments preparation method |
| CN1038813C (en) * | 1993-07-02 | 1998-06-24 | 周勇 | Traditional Chinese medicine for curing osteoporosis |
| CN1368145A (en) * | 2001-02-02 | 2002-09-11 | 杨孟君 | Nano medicine 'Gengnianshu' and its preparing process |
| CN1569017A (en) * | 2003-07-24 | 2005-01-26 | 复旦大学附属华山医院 | Total epimeddium flavone tablet and its preparation method |
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