CN1196472C - Bacterial Chinese medical preparation and preparing method thereof - Google Patents
Bacterial Chinese medical preparation and preparing method thereof Download PDFInfo
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- CN1196472C CN1196472C CNB021576661A CN02157666A CN1196472C CN 1196472 C CN1196472 C CN 1196472C CN B021576661 A CNB021576661 A CN B021576661A CN 02157666 A CN02157666 A CN 02157666A CN 1196472 C CN1196472 C CN 1196472C
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Abstract
The present invention discloses a Chinese medicinal preparation using fungi for abirritation and a preparation method thereof, which belongs to the technical field of Chinese medicinal preparations. Analgesic drugs are prepared after marasmius androsaceus strains are cultured and fermented by way of aerated fermentation. Indicated by pharmacodynamic tests, the Chinese medicinal preparation can increase the basic pain threshold, and has the advantages of long hold time, small side effect and no dependency.
Description
Technical field:
The invention belongs to the Chinese medicine preparation technical field.Be specifically related to a kind of Chinese medicine preparation and preparation method of mycopowder of the mushroom with analgesic activity.
Background technology:
" mushroom Chinese medicine " is a pith of the natural drug precious resources in the Chinese medicine treasure-house.Become exploration in recent years, development has the new drug key areas.As Ganoderma, cloud medicine, Lentinus Edodes, Cordyceps etc. all is the common use of China extremely wide " mushroom Chinese medicine ", has immunomodulating.The anti-ageing drug effect of waiting for a long time, and have only marasmius androsaceus in the mushroom Chinese medicine is unique neuralgic analgesic of all kinds for the treatment of, the kind of each province, city's health, the approval of pharmaceutical control and administration department has been produced old mushroom Chinese medicine of two more than ten years, also is unique medicinal fungi that definite analgesia effect is arranged.So far there is not this kind abroad.The commodity marasmius androsaceus adopts solid fermentation method in the market, for the crude extract of dried mycelium together with culture matrix, with workshop-based production method, forms the mycelia that grows on the culture medium at agricultural and sideline material such as Testa Tritici, Testa oryzae.But this technology labor intensity is big, and changing factor is many, and four months production cycles do not wait by 1 year, do not have the quality control index of science.Once had report to adopt deep layer (liquid) fermentation method for producing, but because of the variation of strain serious degradation, product, quality can not reach the formulation index, so much the factory went bankrupt, the product atrophy can not be satisfied the market demands of modern society.
In the analgesic except strong analgesics to malignant tumor and surgical operation therapy, the neuralgic analgesic of all kinds more commonly, as trigeminal neuralgia, sciatica, migraine, leprosy neuralgia, supraorbital neuralgia, facial paralysis, lumbar muscle strain and rheumatic arthritis wait the patient bitterly, be the frequently-occurring disease of modern society, account for more than 20% of population, especially disease common especially and that heal than refractory in tunnel work and navigation, textile industry.The analgesic (NSAID) of non-steroid type that commonly used is is as aspirin, ibuprofen, and ibuprofens etc. can alleviate pain, " the thunder syndrome " that but causes gastric ulcer and renal function to reduce.At present, both at home and abroad to treating above-mentioned various types of neuralgia the less medicine of toxic and side effects more rare.
Summary of the invention:
Technical problem to be solved by this invention is to overcome above-mentioned defective, design good analgesic effect, the Chinese medicine preparation that side effect is little.
The invention provides a kind of mushroom Chinese medicine preparation, said preparation is that MA-462-A forms as active ingredient and pharmaceutic adjuvant by the Marasmius androsaceus (L.ex Fr.) Fr. mycopowder, and the content of Marasmius androsaceus (L.ex Fr.) Fr. mycopowder and pharmaceutic adjuvant is 100% of total formulation weight amount.Wherein the content of active ingredient Marasmius androsaceus (L.ex Fr.) Fr. mycopowder is to account at least 70% of total formulation weight amount.
Above-mentioned pharmaceutic adjuvant is conventional adjuvant.
The inventor has carried out the animal efficacy testing to Chinese medicine preparation of the present invention (fungus No. 1) and active ingredient, and its result is as follows:
Preparation each several part of the present invention tests for the mice oral administration to mice acetic acid twisting method analgesic activity;
Summary:
1, mice oral administration anluotone tablet is 4.8g/kg/ time, and is 2 times/day * 3 days, after the administration 0.5 hour, 1 hour and 1.5 hours, to the analgesic activity of mice acetic acid twisting method, more remarkable with 1 hour.
2, mice oral administration 4.8g/kg/ time given at No. 1 position of fungus 1～7, and 2 times/day * 3 days, to the test of mice acetic acid twisting method analgesic activity, wherein position 3 analgesic activities were more remarkable.
3, mice oral administration 4.8g/kg/ time given at No. 1 position of fungus 8～13, and 2 times/day * 3 days, to the test of mice acetic acid twisting method analgesic activity, wherein position 9,12 analgesic activities were more remarkable.
Positive drug anluotone tablet and CNNS's effective rheumatism medicine sheet all have analgesic activity to mice acetic acid twisting method with the same dose administration, its analgesic activity no significant difference.
One, after anluotone tablet was given the mice oral administration, different time was measured Dichlorodiphenyl Acetate writhing method analgesic activity
1, test objective:
Measure the analgesic activity of anluotone tablet with mice acetic acid twisting method to different time points behind the mice oral administration.
2, be subjected to the reagent thing
Title: anluotone tablet (positive control drug) Foshan pharmacy one factory
Sample supplier: herbal biomedical engineering institute
Content: 0.24g/ sheet
Preparation: be mixed with the 0.12%/ml suspension with 0.5%CMC-Na
3, animal
Kunming mouse is provided by Shanghai Institute of Pharmaceutical Industry's Animal House, moving No. 107 of Shanghai
Body weight: 18～22g, male and female half and half
Each treated animal number: 10
Dosage is provided with: 4.8g/kg/ time, and 2 times/day * 3 days
Route of administration: per os is irritated stomach
Administration volume: 0.8ml/20g body weight
Test method:
18～22g Kunming mouse, male and female half and half, be divided into 4 groups at random, every group 10, be respectively after negative control group, the administration 0.5,1 and 1.5 hour 3 groups, measure mouse peritoneal inject turn round in 10 minutes behind 0.7% acetic acid body number (stretch hind leg, abdominal part shrinks indent, body distortion simultaneously), that calculates every group of mice on average turns round the contrast of body number and negative control group, carries out data statistics and handles (t check), the calculating % that eases pain.
Result of the test:
The total data statistical procedures is with meansigma methods (X ± SD) expression, with the significance between more following each group of t method of inspection, the result shows after the administration 0.5,1,1.5 hour, and there were significant differences with the negative control group contrast, with after the administration 1 hour more remarkable.See Table 1.
Conclusion (of pressure testing):
Anluotone tablet is given mice oral administration 4.8g/kg/ time, after 2 times/day * 3 days, measures in 0.5,1 and 1.5 hour, and mouse writhing method is had obvious analgesic effect, and 1 hour analgesic activity is more remarkable after the administration.
Two, the analgesic activity of No. 1 each position 1～13 pair of mice acetic acid twisting method of fungus and CNNS's sheet and CNNS's effective rheumatism medicine sheet are to the analgesic activity of mice acetic acid twisting method
1, test objective
Measure anluotone tablet and CNNS's effective rheumatism medicine sheet analgesic activity to mice acetic acid twisting method.
The analgesic activity of No. 1 position of fungus 1～13 pair of mice acetic acid twisting method.
2, be subjected to the reagent thing
Title 1: anluotone tablet Foshan pharmacy one factory
Content: 0.24g/ sheet
Title 2: effective rheumatism medicine sheet Jilin Province Jian Pharmacy stock Co., Ltd of CNNS
Content: 0.6g/ sheet
Title 3: No. 1 each position 1～13 of fungus, totally 13 samples
Preparation: will be mixed with the suspension of 0.12/ml behind the sample porphyrize with 0.5%CMC-Na.
3, animal
Kunming mouse is provided by Shanghai Institute of Pharmaceutical Industry's Animal House, and No. 107 body weight: 18～22g, male and female half and half are moved in Shanghai
Each treated animal number: 5; 10
Dosage is provided with: 4.8g/kg/ time, and 2 times/day * 3 days
Route of administration: per os is irritated stomach
The administration volume; 0.8ml/20g body weight
Test method:
18～22g Kunming mouse, male and female half and half, be divided into 4 groups at random, every group 10, be respectively negative control group, anluotone tablet group, CNNS's effective rheumatism medicine sheet group and No. 1 each position of fungus 1～13 each group, inject for behind the mice oral administration 1 hour and measure mouse peritoneal to turn round in 10 minutes behind 0.7% acetic acid body number (stretch hind leg, abdominal part shrinks indent, body distortion simultaneously), that calculates every group of mice on average turns round the contrast of body number and negative control group, carry out data statistics and handle (t check), calculate analgesia percentage rate (%).
Result of the test:
Total data learn by statistics processing with mean+SD (expression of X ± SD), with the significance between more following each group of t method of inspection, the result shows:
(1) in No. 1 position 1～7 of fungus, the analgesic activity of position 3 pairs of mice acetic acid twistings method is stronger, with negative control group contrast difference highly significant; In position 8～10,11～13, position 9 and 12 analgesic activities are stronger, see Table 2, table 3.
(2) positive drug anluotone tablet, CNNS's effective rheumatism medicine sheet have analgesic activity to mice acetic acid twisting method, and there were significant differences with the negative control group ratio.Anluotone tablet and CNNS's effective rheumatism medicine sheet see Table 3 to the analgesic activity no significant difference of mice acetic acid twisting method.
(3) anluotone tablet mice oral administration is 4.8g/kg/ time, 2 times/day.
Mice acetic acid twisting method has significant analgesia role, sees Table 4.
Discuss:
(1) No. 1 dosage of fungus can be reduced to 4.8g/kg/ day, 2 times/day * 3 days.
(2) CNNS's effective rheumatism medicine sheet is given mice oral administration 4.8g/kg/ time, and 2 times/day * 3 days,
Observe mice have close one's eyes, quiet and movable few behavior, the visible animal of naked eyes becomes thin.
0.5,1,1.5 hour analgesic activity behind No. 1 positive control drug anluotone tablet of the table 1 fungus mice oral administration to mice acetic acid twisting method
Group number of animals (n) is on average turned round body number (X ± SD) analgesia (%)
Negative control 10 23.1 ± 6.2
0.5h 10
**15.4±3.9 33
CNNS
1.0h 10
***13.2±3.3 42.9
Pain sheet 1.5h 10
*16.4 ± 6.0 29.0
*P＜0.05
**P＜0.01
***P＜0.001
No. 1 position of table 2 fungus 1～7 mice oral administration is to mice acetic acid twisting method analgesic activity
Group number of animals (n) is on average turned round body number (X ± SD) analgesia (%)
Negative control 10 21.5 ± 4.8
Anluotone tablet 10
*14.7 ± 7.2 31.6
1 10
**11.4±6.8 47.0
2 10
*14.3±7.1 33.5
Portion 3 10
* *8.2 ± 4.0 61.9
4 10
**14.0±3.6 34.9
Position 5 10 20.6 ± 8.0 4.18
6 10
**16.5±2.6 23.3
7 10 19.1±5.5 11.2
*P＜0.05
**P＜0.01
***P＜0.001
No. 1 position of table 3 fungus 8～13 mice oral administrations are to mice acetic acid twisting method analgesic activity (marasmius androsaceus liquid fermentation mycelium different parts mice oral administration analgesic activity)
The group number of animals is on average turned round the analgesia of body number
(n) ( X±SD) (％)
Negative control 10 21.8 ± 4.4
Anluotone tablet 10
* *13.6 ± 2.4 37.6
CNNS's effective rheumatism medicine sheet 10
* *11.7 ± 3.9 46.3 become thin, quiet effect
8 10
*17.3 ± 3.3 20.6 black thalline alcohol-insoluble substances
Organize 9 10
* *13.1 ± 2.8 39.9 black thalline alcohol-insoluble substances
10 10
*16.5 ± 4.3 24.3 black mycelium
Divide 11 10
* *14.3 ± 4.0 34.4 diphtheria body dissolve with ethanol things
12 10
* *11.2 ± 4.2 48.6 diphtheria body dissolve with ethanol things
13 10
* *12.0 ± 4.1 45.0 diphtheria filaments
*P＜0.05
**P＜0.01
***P＜0.001
No. 1 positive drug anluotone tablet of table 4 fungus mice oral administration (4.8g/kg/ time * 3 times) is to the analgesic activity of mice acetic acid twisting method
Group number of animals (n) is on average turned round body number (X ± SD) analgesia (%)
Negative control 10 24.4 ± 2.7
Anluotone tablet 10
* *14.0 ± 3.2 42.1
*P＜0.05
**P＜0.01
***P＜0.001
Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium extract is tested mice acetic acid twisting method analgesic activity:
Summary:
Mice oral administration Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium extract 1,2,3,4,5, dosage is 2.6g/kg/ day, 2 times/day * 3 days, to the analgesia of mice acetic acid twisting method
The effect test result shows with 5 extracts of negative group contrast all analgesic activity, and wherein the analgesia percentage rate with extract 5 is the highest.
1. test objective
Measure Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium extract with mice acetic acid twisting method
1, the analgesic activity of 2,3,4,5 pairs of mice acetic acid twisting methods.
2. be subjected to the reagent thing
Title: anluotone tablet (positive control drug)
Sample supplier: Shanghai Chinese medicine three factories
Content: 0.12g/ sheet
Title: Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium
Extract 1:95% ethanol extraction, 95% alcohol extraction
Extract 2:95% ethanol extraction, n-butanol extraction
Extract 3:95% ethanol extraction, chloroform extraction
Extract 4:95% ethanol extraction, the water extraction
Extract 5:95% ethanol extraction
Sample supplier: herbal biomedical engineering institute
Preparation: with a small amount of tween 80 hydrotropy, reuse 0.5%CMC-Na is mixed with the 0.037g/ml suspension
3. animal
Kunming mouse is provided by Shanghai Institute of Pharmaceutical Industry's Animal House, and No. 107 body weight: 18～22g, male and female half and half are moved in Shanghai
Each treated animal number: 10
Dosage is provided with: 2.6g/kg day, 2 times/day * 3 days
Route of administration: per os is irritated stomach
The administration volume; 0.7ml/20g body weight
4. test method:
18～22g Kunming mouse, male and female half and half are divided into 8 groups at random, and 10 every group,
Be respectively negative control group, positive controls, extract 1,2,3,4,5 each group, inject for behind the mice oral administration 1 hour and measure mouse peritoneal to turn round in 10 minutes behind the 0.7% acetic acid 0.21ml body number (stretch hind leg, abdominal part shrinks indent, body distortion simultaneously), that calculates every group of mice on average turns round the contrast of body number and negative control group, carry out data statistics and handle (t check), calculate analgesia %.
5. result of the test:
Total data is learned processing by statistics, and (X ± SD) expression is with the significance between more following each group of t method of inspection with mean+SD.
The result shows: Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium and extract 1,2,3,4,5 each group all have analgesic activity to mice acetic acid twisting method.
6. discuss:
Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing mycelium and extract 1,2,3,4,5 all have analgesic activity, and same there were significant differences than them with negative control group, and its analgesic percentage rate is higher than anluotone tablet.
The analgesic activity of table 5 marasmius androsaceus submerged fermentation mycelium and 1,2,3,4,5 pairs of mice acetic acid twisting methods of extract
Group dosage (g/kg/ day) number of animals (n) is on average turned round body number (X ± SD) analgesia (%)
Negative control 0.7ml/20g 10 20.5 ± 4.8
Anluotone tablet 2.6 10
*13.4 ± 3.5 34.6
Extract 1 2.6 10
*13.1 ± 4.9 36.0
Extract 2 2.6 10
* *11.7 ± 4.1 42.9
Extract 3 2.6 10
* *10.1 ± 3.3 50.7
Extract 4 2.6 10
* *12.2 ± 4.5 40.5
Extract 5 2.6 10
* *9.7 ± 4.0 52.7
Mycelium 2.6 10
* *10.3 ± 4.8 49.8
**P＜0.01，
***P＜0.001
Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation product is tested mice acetic acid twisting method analgesic activity
Summary:
One, mice oral administration Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation thing extract crystallization K
1, dosage is 2.1g/kg/ day, 2 times/day totally 3 days, to the analgesic activity test of mice acetic acid twisting method, the result showed that with the negative control group contrast significant analgesia role, analgesia rate being arranged is 50.7%.
Two, mice oral administration Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation mycelium 1 and 2, dosage is respectively 2.6g/kg/ day, 2 times/day totally 3 days, the analgesic activity test result of mice acetic acid twisting method is shown that with the negative control group contrast significant analgesia role is arranged.
1. test objective:
Measure Marasmius androsaceus (L.ex Fr.) Fr. deep-fermentation mycelium 1 and 2, submerged fermentation extract crystallization K
1Analgesic activity to mice acetic acid twisting method.
2. be subjected to the reagent thing:
Title: anluotone tablet (positive control)
Sample supplier: Shanghai Chinese medicine three factories
Content: 0.12g/ sheet
Title: Marasmius androsaceus (L.ex Fr.) Fr. submerged fermentation mycelium 1
Marasmius androsaceus (L.ex Fr.) Fr. submerged fermentation mycelium 2
Marasmius androsaceus (L.ex Fr.) Fr. submerged fermentation extract crystallization K
1
Sample supplier: herbal biomedical engineering institute
Preparation: the sample porphyrize is mixed with the suspension of 0.037g/ml with 0.5%CMC-Na
3. animal:
Kunming mouse is provided by Shanghai Institute of Pharmaceutical Industry's Animal House, and No. 107 body weight: 18～22g, male and female half and half are moved in Shanghai
Each treated animal number: 10
Dosage is provided with: 2.6g/kg/ day, and 2 times/day totally 3 days,
2.1g/kg/ day, 2 times/day totally 3 days
Route of administration: per os is irritated stomach
Administration volume: 0.6ml/20g body weight
4. test method:
18～22g Kunming mouse, male and female half and half are divided into 5 groups at random, 10 every group, are respectively negative control group; 1 group of positive controls, mycelium, 2 groups, extract crystallization K
1Group.1 hour pneumoretroperitoneum of mice oral administration is injected 0.6% acetic acid, and (stretch hind leg, abdominal part shrinks concavo-convex to measure the body number of turning round in 10 minutes, body distortion simultaneously), that calculates every group of mice on average turns round the contrast of body number and negative control group, carries out data statistics and handles (t check), the calculating % that eases pain.
Analgesia (%)=(negative control group is turned round the several administration groups of body and turned round the body number)/negative control group is turned round body and is counted * 100%
5. result of the test:
Total data is learned processing by statistics with mean+SD (X ± SD) expression, the significance between relatively each is organized with the t method of inspection.The result shows: extract crystallization K
1Analgesic activity is all arranged, extract crystallization K with 1,2 pairs of mice acetic acid twisting methods of mycelium
1Analgesia rate the highest (seeing Table 6).
6. conclusion:
The analgesia rate of Marasmius androsaceus (L.ex Fr.) Fr. mycelium 1,2 is identical with anluotone tablet, crystallization K
1The analgesia rate be 50.7%.
Table 6 marasmius androsaceus submerged fermentation mycelium and extract K
1Analgesic activity to mice acetic acid twisting method
Group dosage (g/kg/ day) number of animals (n) is on average turned round body number (X ± SD) analgesia (%)
Negative control 0.7ml/20g 10 24.5 ± 4.8
Anluotone tablet 2.6 10 15.1 ± 4.6
* *34.6
Mycelium 1 2.6 10 15.0 ± 4.3
* *36.0
Mycelium 2 2.6 10 17.3 ± 5.6
* *42.9
Crystallization K
12.1 10 13.6 ± 3.4
* *50.7
Annotate:
*P＜0.01,
* *P＜0.001
The technical characterictic of the invention is:
1. use biofermentation technique, find out the spawn degeneration reason, illustrating when marasmius androsaceus MA-462 grows on agar culture medium has four kinds of cell growth patteries, the speed of growth of every kind of cell growth pattern, and morphological characteristic is all different, fermentating metabolism product difference when entering liquid fermentation, the extract difference, and the analgesia drug effect is also different, the bacterial strain of four kinds of cell growth patteries, its ethanol extraction output maximum differs more than four times, and analgesia drug effect maximal phase differs from more than two times.For this reason, the bacterial strain of therefrom choosing a kind of specific good growth pattern is as producing strain MA462-A, guarantee strain stability and product quality homogeneity, improved analgesia effect, shortened fermentation period, thereby changed the wild resource rareness of cuticle gill fungi, the domestication cultivates difficulty and solid fermentation mycelia production cycle long (more than half a year), and fermentation impurity is many, yields poorly, there is not quality control index, shortcoming such as alcohol consumption is big.
2. the pharmacology and the biological activity combination test of each composition have systematically been studied, show that the bioactive substance that is had in the mycelium powder of marasmius androsaceus is not single chemical compound, by analytical methods such as GC-MS more than ten kind of organic acid and sterol are arranged really, chemical compounds such as flavonoid all have analgesic activities.The analgesia drug effect of the mycelium powder of liquid fermentation is better than existing market product marasmius androsaceus, and therapeutic dose is equivalent to the 1/20 mycelia amount of present product.Product is that dried mycelium is made marasmius androsaceus together with the crude extract of substrate at present, and the analgesia composition is pure dissolubility, and every tablet preparation is 1.2 gram alcohol extracts, and the mycopowder analgesia drug effect of 1.2 grams is better than 1.2 gram ethanol extract.So adopting mycelium powder is the raw material of clinical application.
3. isolating ergosterol and adenosine from mycelium powder is reference compound, is authenticating compound with the flavone compound, all is the neural spasmolysis of analgesic activities material and moderate according to documents and materials and pharmacodynamics proof.
Another object of the present invention has provided the preparation method of above-mentioned mushroom Chinese medicine preparation, and this method comprises the following steps:
(1) bacterial strain:
Marasmius androsaceus Marasmius androsaceus (L.Fr) Fr available from Edible Fungus Inst., Shanghai Academy of Agriculture, sees Edible Fungus Inst., Shanghai Academy of Agriculture's " edible fungus species catalogue " 1993 Chinese editions.
(2) cultivation of bacterial strain:
At first with bacterial classification inoculation in the culture dish to shaking bottle, cultivate back subcultivation to 50～100L first class seed pot through rotary shaking table, and then expand 500～1000L secondary breeding jar to, change 500L～10 ton three grade fermemtation jar at last over to;
Seed tank, the inventory of breeding jar and fermentation tank is 70%, inoculum concentration is 10%～15%, 23～30 ℃ of jar temperature, 1: 0.3～0.5v/v of ventilation, speed of agitator 50～200r.p.m, fermentation nitrogen source is without peptone and yeast extract, and carbon source is without maltose, but selects soybean cake powder and glucose for use, the incubation time of seed tank and breeding jar is 48～72 hours, and the fermentation time of fermentation tank is 48～60 hours;
(3) Marasmius androsaceus (L.ex Fr.) Fr. mycopowder preparation:
The fermentation tank breeding is after 48～60 hours, mash is through plate-and-frame filtration or centrifuging elimination mash, precipitate (wet mycelium) put to 70～80 ℃ of drying rooms after the ventilation drying, be ground into mycopowder, formulation method incapsulates mycopowder or mycopowder is made thin membrane coated tablet routinely then.
Marasmius androsaceus powder producing method of the present invention can obtain the mycopowder of highly active analgesia drug effect, and following characteristics and practical function are arranged:
1, marasmius androsaceus powder producing method of the present invention is a strain with Chinese marasmius androsaceus MA462-A, adopts the aerobic fementation method, can cultivate a large amount of mycopowder in 2-3 days, fast 90～180 times than immobilized solid fermentation method.
2, the present invention adopts mycopowder to compare with marasmius androsaceus tablet in the market as curative increases by 20 times of output and cancels a large amount of 95% ethanol that (the marasmius androsaceus tablet is that the mycelial 95% ethanol crude extract of solid fermentation 1.2 grams get through the 95% ethanol extraction face of 400～450ml at present, and zooperal analgesia drug effect is lower than the mycelium powder (mycopowder) of liquid fermentation.
3, mycopowder of the present invention contains multiple analgesic activities material, so have the synergism of analgesia drug effect.
4, mycopowder of the present invention has two reference compounds and authenticating compound and physicochemical property thereof, structural formula, stability, the quality control index of content.
The analgesia drug effect of liquid fermentation mycopowder of the present invention and existing market product marasmius androsaceus coated tablet and their production method are compared as follows:
| Production method | The product tablet is formed and weight | The required mycelia amount of ethanol extraction | Amount of alcohol | The analgesia suppression ratio |
| Solid fermentation | Mycopowder | Agricultural and sideline thing | Ethanol extraction | Mycelia amount (gram)/1.2 gram extract | 95% (ml)/1.2 gram extract | Five crowdes of average % |
| Solid fermentation | 1.2 | 1.2 | 18～20 grams | 400～450 | 30～35％ | |
| Liquid fermentation | 1.2 | / | / | / | / | 36～50％ |
Useful result of the present invention:
1. the technical characterictic of above relevant the invention is the special physiological characteristic of the peculiar rare medicinal fungi marasmius androsaceus of China, do not appear in the newspapers so far both at home and abroad, and be characteristic Chinese crude drug new resources.
2. adopt biotechnology, the application and development of the modernization of Chinese medicine engineering of the new quality control of new production process of submerged fermentation technology, the modernized Chinese crude drug that the ancient and precious analgesic of producing for two more than ten years is increased technology content is widened the thinking and the research and development of new Chinese medicine exploitation.
It is long that this medicine is kept analgesic time, and pain relief is to disappearing thereupon, and its analgesia effect is through long-term, a large amount of clinical practices, had vast showing in market, after the application and development by modernization project, enlarging at produce market will be convenient, and old Chinese medicine is called forth one's youthful vigor.
The specific embodiment:
Embodiment 1,
Take liquid nitrogen and be kept at Chinese marasmius androsaceus MA462-A and be implanted on the PDA-B slant medium, cultivated 7～10 days for 24～27 ℃, the PDA-B prescription is as follows:
Rhizoma Solani tuber osi liquor 960ml
Glucose 20g
Husband's skin 1～10g
pH 6～6.5
Peeling Rhizoma Solani tuber osi 200g adds water boil extracting juice after 5 minutes, sterilization: 1.2kg/cm
230 minutes.
Get slant strains, Chinese Marasmius androsaceus (L.ex Fr.) Fr. mycelium immigration is equipped with in the 500ml triangular flask of 100ml fluid medium, 24～26 ℃ of rotary shakers 4 days merge the shake-flask culture thing, " pressure differential method moves and inserts jar interior loading amount 70% (v/v) in 50～150L seed tank, and inoculum concentration 10% (v/v) aerobic culture moved into 500L after two days
3～1000L
3Breeding jar, treat mycelia long dense after again with 1: 10 volume ratio expanding propagation to fermentation tank, 24～30 ℃ of jar temperature, ventilation 1: 0.2～1.0 (v/v), mechanical agitation speed 20～200rpm.
Fermentative medium formula is:
Glucose 40g
Semen Maydis pulp 2g
KH
2PO
41.2g
Soybean cake powder 5g
OXY 3g
MgSO
4 1.0g
VB
1 100μg
Add water 1000ml
The fermentation tank breeding is after 48～60 hours, and mash gets the mycelium filter cake through plate-and-frame filtration, and separately frustillatum advances drying room, ventilation, and 70～80 ℃, oven dry is ground into mycopowder, for the brown powder of Semen Armeniacae Amarum fragrance is arranged.
Embodiment 2,
Get the Chinese marasmius androsaceus MA-462A bacterial strain under the Dormant oils preservation, move and grow, carry out activation culture in the GMY culture medium, the GMY culture medium prescription.
Glucose 10g
Yeast decoction 4g
Beerwort (pol 16) 100ml
Water fills up 1000ml
The PH nature
Agar 20g
Adorn in the 250ml triangular flask of 50ml culture medium in will moving into the mycelium of activatory Chinese marasmius androsaceus MA-462A on the GMY culture medium, cultivated 3 days with rotary shaking table machine, rotating speed 120～180rpm, enlarging inoculation subsequently goes in the seed tank, ventilation 1: 0.2-1 (v/v) cultivates after 2～3 days for 23～30 ℃ can move into fermentation tank, fermentation culture, scale is with embodiment 1.
Fermentative medium formula is as follows:
Glucose is 3%
Soybean cake powder 1%
MgSO
4 0.05％
Glycerol 1%
K
2HPO
4 0.1％
The PH nature
Semen Maydis pulp 0.5%
KH
2PO
4 0.1％
Behind the aerobic fementation 2～3 days,, wet mycelium is milled to by 100 purpose mycopowder by embodiment 1, is brown powder, distribute special fruit fragrance with centrifuging elimination mash.
The method of quality control of above mycopowder (embodiment 1,2).
1, surveys reference compound
Detect two reference compounds with the high-pressure liquid phase method
Gland salidroside content＞0.4%
Ergosterol＞0.5%
2, detect an authenticating compound flavone compound with the thin plate layer analysis method and have speckle.
Embodiment 3, mycopowder preparation process one thin membrane coated tablet
1. raw material (mycopowder), adjuvant → mixing → granulation → drying → granulate → tabletting
2. art for coating:
Get polyvinylpyrrolidonesolution solution and raw material and adjuvant combination drying, the granulate tabletting, get filmogen, be dissolved in the ethanol, sieving with the Pulvis Talci material adds the back mixing, food coloring is dissolved in the boiling water, it is standby to add above-mentioned solution mixing, then, and the pressure powder that sieves, make it dry with the heating of nebulization limit, homogeneous film gets final product.
3. coating disintegrate speed per hour half an hour
4. preparation (thin membrane coated tablet) adjuvant
Mycopowder 500mg
Microcrystalline Cellulose CH102 120mg
Lactose 40mg
CaSO
4 40mg
CMSNA (sodium carboxymethyl) 40mg
Magnesium stearate 4mg
Do binding agent with 15%PVP k30, the preparation of 70% ethanol.
Embodiment 4, mycopowder preparation process-capsule
Fill No. 1 capsule (meeting pharmacopeia) of packing into mycopowder (meeting quality standard) 0.4 gram.
Claims (2)
1, a kind of preparation method of Chinese medicine preparation of mycopowder of the mushroom with analgesic activity is characterized in that this method comprises the following steps:
(1) bacterial strain:
Marasmius androsaceus Marasmius androsaceus available from Edible Fungus Inst., Shanghai Academy of Agriculture, sees Edible Fungus Inst., Shanghai Academy of Agriculture's " edible fungus species catalogue " 1993 Chinese editions;
(2) cultivation of bacterial strain:
At first with bacterial classification inoculation in the culture dish to shaking bottle, cultivate back subcultivation to 50～100L first class seed pot through rotary shaking table, and then expand 500～1000L secondary breeding jar to, change 500L～10 ton three grade fermemtation jar at last over to;
Seed tank, the inventory of breeding jar and fermentation tank is 70%, inoculum concentration is 10%～15%, 23～30 ℃ of jar temperature, 1: 0.3～0.5v/v of ventilation, speed of agitator 50～200r.p.m, fermentation nitrogen source is without peptone and yeast extract, and carbon source is without maltose, but selects soybean cake powder and glucose for use, the incubation time of seed tank and breeding jar is 48～72 hours, and the fermentation time of fermentation tank is 48～60 hours;
(3) Marasmius androsaceus (L.ex Fr.) Fr. mycopowder preparation:
After the fermentation tank breeding 48～60 hours, mash is through plate-and-frame filtration or centrifuging elimination mash, precipitate put to 70～80 ℃ of drying rooms after the ventilation drying, be ground into mycopowder, and formulation method incapsulates mycopowder or mycopowder is made thin membrane coated tablet routinely then.
2, a kind of Chinese medicine preparation of mycopowder of the method preparation by claim 1, it is characterized in that said preparation is made up of as active ingredient and pharmaceutic adjuvant the Marasmius androsaceus (L.ex Fr.) Fr. mycopowder, wherein the content of Marasmius androsaceus (L.ex Fr.) Fr. mycopowder and pharmaceutic adjuvant is 100% of total formulation weight amount, and the content of active ingredient Marasmius androsaceus (L.ex Fr.) Fr. mycopowder is to account at least 70% of total formulation weight amount.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021576661A CN1196472C (en) | 2002-12-23 | 2002-12-23 | Bacterial Chinese medical preparation and preparing method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021576661A CN1196472C (en) | 2002-12-23 | 2002-12-23 | Bacterial Chinese medical preparation and preparing method thereof |
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| CN1424054A CN1424054A (en) | 2003-06-18 |
| CN1196472C true CN1196472C (en) | 2005-04-13 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100353828C (en) * | 2004-06-04 | 2007-12-12 | 毛福森 | Composite fungus product for raising human body immunity and its prepn process |
| CN102579509B (en) * | 2012-01-18 | 2014-05-14 | 山西康欣药业有限公司 | Selenium-enriched marasmius androsaceus toadstool extract preparation and preparation method thereof |
| CN104857027B (en) * | 2015-04-19 | 2019-01-18 | 吉林大学 | CNNS Marasmius T08 is preparing the medical application in analgesic |
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