CN101647811A - Bionic enzymatic product of animal medicament and application thereof - Google Patents
Bionic enzymatic product of animal medicament and application thereof Download PDFInfo
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Abstract
The invention discloses a bionic enzymatic product of animal medicament and application thereof, which belong to the field of Chinese medicament. The bionic enzymatic product is characterized in thata bionic enzymatic method is adopted, and comprises the following steps: taking animal medicament; grinding the animal medicament into fine powders; adding water into the fine powders; evenly stirringthe mixture; under appropriate condition, performing heat preservation by using pepsin; performing heat preservation by using pancreatin or trypsase; and preparing obtained zymolyte into preparationsaccording to different preparation requirements. The product of the invention has good effects in the aspects of resisting hepatitis virus, reducing blood press and resisting cancer.
Description
Technical field
The present invention relates to a kind of bionic enzymatic hydrolysate of animal drugs and in the application of hepatitis virus resisting, blood pressure lowering and anticancer aspect.Belong to the field of Chinese medicines.
Technical background
Animal tcm has very long history in the application of China.Before more than 3000 years, China has just begun the utilization to Apis.And the breed of Margarita, male bat also sees China the earliest, the history in existing more than 2000 year.China's herbal books Shennong's Herbal the earliest records 365 kinds of medicines, and wherein animal drugs just has 65 kinds; According to incompletely statistics, in " Book of Songs " book, recorded about 160 kinds of all kinds of animals altogether, both edibles of many animals have been arranged, but also hyoscine; In addition, the Classic of Mountains and Rivers of the Spring and Autumn and the Warring States Periods has also recorded 67 kinds of medicinal animals; The Shennong's Herbal in Qin Han period records animal drugs 65 flavors; " Newly Revised Canon of Materia Medica " of the Tang Dynasty records animal drugs 128 flavors; Ming Dynasty's Compendium of Material Medica is recorded animal drugs 461 flavors, and it is divided into worm, squama, Jie, fowl, beast, Ren Gebu; Qing Dynasty's ZHAO Xue-Min supplementary Amplifications of the Compendium of Materia Medica records animal drugs 128 flavors; " the Chinese medicine voluminous dictionary " in modern age records nearly 740 flavors of animal drugs; According to modern book on Chinese herbal medicine record, kind surplus the animal drugs of land and ocean closely reaches 1600.About 300 pleasant impressions of Chinese crude drug that tcm clinical practice is commonly used, wherein animal drugs accounts for 10%.With 2000 editions pharmacopeia is example, records Chinese medicine 534 flavors altogether, and animal drugs 47 flavors account for 8.8%; Compound recipe or folk prescription 458 sides contain animal drugs prescription totally 153 sides, account for 33.4%, this shows that in the bed medicinal herbs most in use, though the animal drugs proportion is less, applicating frequency is high, show that animal drugs occupies important position in clinical practice; And the clinical practice determined curative effect of animal drugs, as effective prescription of clinical practice have that cow-bezoar bolus for resurrection, restorative bolus, plum blossom tongue ball, purple snow, WUJI BAIFENG WAN, Anisodus carniolicoides C.Y.Wu et C.Chen, wanying troche, Zhibao Dan, Bezoar pill for lowering blood pressure, Bezoar Sedative Pills, cow-bezoar anti-toxic bolus, longevity powder, office side's most valuable treasure loose, Fel Serpentis et Bulbus Fritillariae Cirrhosae Liquidus, QINGKAILING etc.Yet a lot of commonly used animal drugs are directly taken after adopting crude drug to pulverize on the mode of being used as medicine usually or water is taken after carrying, and using method is tradition relatively, and the medical material utilization rate is lower, and effective ingredient does not obtain to make full use of.Through the modern times comparatively biological activity relevant with clinical efficacy and the material base evaluation study of system, the curative effect of water-solubility protein in the animal drugs and peptide class, free aminoacid ingredient more and more receives publicity.After the human body oral administration animal drugs, enzymolysis, digestion through pepsin and pancreatin, be absorbed into blood performance curative effect with the small-molecular peptides constituents, micromolecular oligopeptides material and aminoacid are easy to by little intestinal absorption, and macromolecular protide also is difficult to be absorbed in small intestinal.Traditional former powder is directly taken, high molecular weight protein in the former medicated powder, have only on a small quantity and decompose through gastrointestinal in vivo, become micromolecular oligopeptide and be absorbed, because the interference of the fineness of pulverizing medicinal materials, the variation of gastrointestinal tract pH and oral other foods causes the incomplete of hydrolysis, what effective ingredient was absorbed and used lacks, and causes the waste curative effect of a large amount of medical materials to reduce greatly.The extraction process that water is carried makes has the albumen of most of insoluble entry to be removed by filtration, causes a large amount of medical material wastes, and curative effect reduces equally greatly.
Use the albumen in the enzymatic isolation method hydrolysis of animal medicine, the biologically active peptide that acquisition has certain physiologically active more and more is subjected to people's attention.Because the position of different enzyme effects is different, the product that enzymatic isolation method obtains is also different, come enzymolysis with single pepsin or pancreatin, can only make the protein part enzymolysis in the medical material, albumen in the medical material can not get maximum utilization, like this with regard to a kind of bionical enzyme solution of exigence---in animal or human's body biological evolution process, be proved, the bionic enzymatic method of the digestion process of determined curative effect, simulation human body, with the easier absorption of the effective ingredient that obtains behind the animal drugs enzymolysis, safer utilization makes full use of crude drug simultaneously more.
Summary of the invention
First purpose of the present invention is to provide a kind of more effective, the bionic enzymatic hydrolysate for animal medicaments of safety; Second purpose of the present invention provides this product in the application that is used for preparing hepatitis virus resisting, blood pressure lowering and anticancer medicine.
The inventor provides a kind of enzymatic hydrolysate of animal drugs, and this product adopts the method preparation of bionic enzymatic:
Get animal drugs, be ground into fine powder, add water, regulate pH to 1.0～3.0, add pepsin in 35～45 ℃ of insulation enzymolysis 0.5～4.0 hour, re-adjustment pH to 7.5～8.5 add pancreatin or trypsin in 40～50 ℃ of insulation enzymolysis 2～8 hours, promptly.
Further be optimized for:
Get animal drugs, be ground into fine powder, add water homogenate, regulate pH to 1.5～2.5,0.5%～5% the pepsin that adds the animal dose was in 35～45 ℃ of insulation enzymolysis 1～3 hour, re-adjustment pH to 7.5～8.5 add 0.5%～5% pancreatin of animal dose or trypsin in 40～50 ℃ of insulation enzymolysis 3～6 hours, promptly.
More excellent preparation method is:
Get animal drugs, be ground into fine powder, add water homogenate, regulate pH to 2.0,1%～2% the pepsin that adds the animal dose is in 40 ℃ of insulation enzymolysis 1～3 hour, re-adjustment pH to 8.0, add 1%～2% pancreatin of animal dose or trypsin in 50 ℃ of insulation enzymolysis 3～6 hours, promptly.
The inventor after the animal drugs fine powder adds water homogenate, can be heated to 80～100 ℃ in advance through experiment sieving, be incubated 15～30 minutes, and it is temperature required to be cooled to enzymolysis again, presses above operation enzymolysis, and its effect is better.Carry out enzymolysis by technical solution of the present invention, when pepsic enzyme activity is not less than 1200U/g, tryptic enzyme activity is not less than 2500U/mg, and the casein conversion power of pancreatin was not less than 25.0 o'clock, and effect is comparatively abundant; Enzyme activity is high more, and enzymolysis speed is fast more, effect is good more.
Prove that through modern study the peptide constituents in the animal drugs also is an effective site.After the human body oral administration animal drugs,, absorb and the performance curative effect with the small-molecular peptides constituents through pepsin and pancreatin or tryptic enzymolysis, digestion.The inventor is according to the process of the oral animal drugs of human body, creationary method at external employing simulation of human body enzymolysis, successively raw material is carried out pepsin, trypsin or pancreatin enzymolysis, compare with the former powder of the direct oral medical material of human body, the active substance faciation that gained is assimilated by small intestinal is same, but external enzymolysis is selected more excellent experimental condition, enzymolysis more abundant, the drug effect of oral gained is stronger again, enzymolysis becomes oligopeptide or micromolecular active site group easier being absorbed when administrations such as injection or mucosa, skin, immunogenicity is lower, and is more effective.Through experiment sieving, separately with pepsin, trypsin, pancreatin, neutral protease, alkaline protease, papain enzymolysis animal drugs, effect all is better than directly oral, but all be lower than bionic enzymatic method of the present invention, the inventive method has strengthened the curative effect of animal drugs such as animal drugs to a great extent, and owing to be used as medicine with the form of micromolecule oligopeptide, oral administration be absorb more abundant, the performance curative effect is rapid, heightens the effect of a treatment, and is worthy of popularization.
Those skilled in the art can cooperate enzymatic hydrolysate of the present invention with suitable adjuvant easily, are prepared into various conventional formulations, as:
A, with the direct spray drying of enzymolysis solution, add an amount of conventional adjuvant such as starch, lactose, microcrystalline Cellulose, carboxymethyl starch sodium etc. and make capsule or tablet.
B, enzymolysis solution filtered be condensed into thick paste, add an amount of Icing Sugar and dextrin and make granule.
C, with enzymolysis solution be heated to 85 ℃ kill enzyme after, filter, filtrate is divided and is got the following solution of molecular cut off 5kD with the ultrafilter membrane ultrafiltration, can add isoosmotic adjusting agent, pH regulator agent, antiseptic etc. and make little pin or transfusion; Also can add lyophilizing such as mannitol, lactose and make lyophilized injectable powder.
D, enzymolysis solution is filtered, filtrate is with the ultrafilter membrane ultrafiltration, the solution that molecular cut off 10kD is following, or filtrate directly adds conventional pharmaceutic adjuvants such as carbomer, chitosan, makes gel or spray.
Animal drugs enzymatic hydrolysate provided by the invention can use separately, also can unite use, that is: in active constituents of medicine, can have only this product with the other drug composition, can also be the mixture of itself and other drug, reach the purpose of partner treatment, auxiliary treatment.
Animal drugs enzymatic hydrolysate of the present invention can extensive use in hepatitis virus resisting, blood pressure lowering, anticancer medicine; Also can be used for the treatment of aspects such as blood circulation promoting and blood stasis dispelling, dispelling wind for relieving itching, dispelling wind and removing obstruction in the collateral pain relieving, heat-clearing and toxic substances removing, cooling blood for hemostasis, antiinflammatory, infection, antiviral, arresting convulsion, analgesia, can be used for the epidemic febrile disease hyperpyrexia, unconsciousness and delirium is sent out the speckle dermexanthesis, hematemesis and epistaxis, infantile convulsion, demented, analgesia, calmness, antiviral etc., as the heating that flu causes, the epistaxis that heat in blood causes, purpura, epilepsy, acute infantile convulsion.
Beneficial effect
For further verifying the therapeutical effect of product of the present invention, the inventor has carried out the animal pharmacodynamic experiment, and " the bionic enzymatic group " in the experiment is the corresponding animal drugs enzymatic hydrolysate that makes by technical solution of the present invention:
One, the drug effect comparative test of antivirus action
1, medicine and reagent
For the preparation of test agent enzymolysis group not: water intaking Cornu Bovis seu Bubali medical material fine powder (80 order) 50g, adds 10 times of amount normal saline, homogenate 30 minutes stirs evenly, and gets 1/5 amount, microporous filter membrane (0.45 μ m) filtration, promptly;
Pepsin enzymolysis group: get 1/5 homogenate, add 1% pepsin of Cornu Bubali medical material amount, regulate pH to 2.0 simultaneously, temperature is 40 ℃ ± 2 ℃, and enzymolysis 4h is stirred in insulation simultaneously, and 85 ℃ of insulation 20min are put coldly, and microporous filter membrane (0.45 μ m) filters, promptly;
The trypsin digestion group: get 1/5 homogenate, add 1% trypsin of Cornu Bubali medical material amount, regulate pH to 8.0 simultaneously, temperature is 50 ℃ ± 2 ℃, and enzymolysis 4h is stirred in insulation simultaneously, and 85 ℃ of insulation 20min are put coldly, and microporous filter membrane (0.45 μ m) filters, promptly;
Pancreatin enzymolysis group: get 1/5 homogenate, add 1% pancreatin of Cornu Bubali medical material amount, regulate pH to 8.0 simultaneously, temperature is 50 ℃ ± 2 ℃, and enzymolysis 4h is stirred in insulation simultaneously, and 85 ℃ of insulation 20min are put coldly, and microporous filter membrane (0.45 μ m) filters, promptly;
Bionic enzymatic group: get 1/5 homogenate, add 1% pepsin of Cornu Bubali medical material amount, regulate pH to 2.0 simultaneously, temperature is 40 ℃ ± 2 ℃, enzymolysis 4h is stirred in insulation simultaneously, transfers pH to 8.0 then, adds 1% trypsin of Cornu Bubali medical material amount, temperature is 50 ℃ ± 2 ℃, enzymolysis 4h is stirred in insulation simultaneously, and 85 ℃ of insulation 20min are put cold, microporous filter membrane (0.45 μ m) filters, promptly;
Other gets Formica fusca medical material fine powder (80 order) 50g, supplies test agent by above-mentioned Cornu Bubali for the preparation of test agent preparation method.
Hyclone; DMEM culture medium (Gibco company product); The HBeAg detection kit; The human liver cancer cell cell strain HepG of HBV-DNA clone transfection
2-2.2.15.
2, test method
The human liver cancer cell HepG of HBV-DNA clone transfection
2-2.2.15 cell culture is in the DMEM culture fluid that contains 10% hyclone and two anti-(100U/mL penicillin and 100 μ g/mL streptomycins), in 37 ℃, 5%CO
2Cultivate under the condition.The take the logarithm cancerous cell of trophophase is diluted to 5 * 10 with the DMEM culture fluid that contains 10% hyclone
7/ mL single cell suspension is inoculated in 96 well culture plates, every hole 100 μ L, and abandoning supernatant behind the adhere-wall culture 24h adds the different volumes medicine, and sample dissolves the mother solution that is mixed with 1mg/mL with 3% dimethyl sulfoxine, and adding DMEM culture fluid to final volume is 200 μ L.Parallel 6 holes of each concentration, matched group adds isopyknic DMEM culture fluid.Draw culture plate supernatant 5 μ L, press HBeAg detectable cassette method and detect the antigenic absorbance of e (A) value, be calculated as follows cell proliferation inhibition rate.
Cell inhibitory rate=(the A value of the A value/control cells of 1-dosing cell) * 100%
3, the concrete result of the test of result of the test sees the following form.
Table 1 Cornu Bubali supplies test agent to HepG
2-2.2.15 cell inhibiting rate is table as a result
| Group | Suppression ratio (%) |
| Enzymolysis group not | ??62.44* |
| Pepsin enzymolysis group | ??66.13* |
| The trypsin digestion group | ??72.40* |
| Pancreatin enzymolysis group | ??73.22* |
| The bionic enzymatic group | ??79.84** |
Annotate: compare * P＜0.05, * * P＜0.01 with matched group.
Table 2 Formica fusca supplies test agent to HepG
2-2.2.15 cell inhibiting rate is table as a result
| Group | Suppression ratio (%) |
| Enzymolysis group not | ??63.32* |
| Pepsin enzymolysis group | ??68.01* |
| The trypsin digestion group | ??75.33* |
| Pancreatin enzymolysis group | ??74.47* |
| The bionic enzymatic group | ??80.13** |
Annotate: compare * P＜0.05, * * P＜0.01 with matched group.
Above-mentioned result of the test shows, more than the bionic enzymatic hydrolysate of two kinds of animal drugs all have stronger anti-HBV effect, with do not add matched group that medicine only adds equal-volume DMEM culture fluid relatively, drug level at 10 μ g/mL demonstrates significant difference, effect than other single enzyme enzymatic hydrolysate is strong, points out bionic enzymatic hydrolysate of the present invention to have antivirus action preferably.
Two, to the hypotensive effect of Hypertensive Rats
1 material
1.1 animal SD rat is provided by Shandong University's Experimental Animal Center.
1.2 medicine
The preparation of enzymolysis mother solution: get Pheretima medical material fine powder (100 order) 100g, add 10 times of amount normal saline, homogenate 20 minutes stirs evenly, promptly;
Enzymolysis solution not: get 1/4 amount enzymolysis mother solution, microporous filter membrane (0.45 μ m) filters, promptly;
Bionic enzymatic liquid: get 1/4 amount enzymolysis mother solution, add 1% pepsin (enzyme activity is 1200U/g), regulate pH to 2.0 simultaneously, 40 ℃ of insulated and stirred enzymolysis 4h transfer pH to 8.0 then, adding 1% trypsin enzyme activity is 2500U/mg), 50 ℃ of insulated and stirred enzymolysis 4h, 85 ℃ of insulation 20min are put cold, microporous filter membrane (0.45 μ m) filters, promptly;
Other gets Bombyx Batryticatus medical material fine powder (80 order) 100g, supplies the preparation method of test agent to prepare test sample by above-mentioned Pheretima.
Blank: normal saline.
Positive controls: the sharp 10g of card taking Top is suspended to 200ml with water.
1.3 the reagent pepsin is available from traditional Chinese medicines group chemical reagent company limited, lot number: F20070914; Trypsin is available from traditional Chinese medicines group chemical reagent company limited, lot number: F20071228; Injection normal saline: specification: 250ml, lot number: 02110504, Zhiying Pharmaceutical Factory, Shenyang produces.
2, method and result
2.1 modeling method is got 70 of healthy male rats, body weight 180～220g, be divided into 7 groups at random, every group 10: the bionic enzymatic sample sets, enzymolysis sample group not, the normal control group, the positive drug group, the blank group, except that the normal control group, all the other each groups adopt the electric shock vola as single stressor rat to be carried out stress stimulation, stimulus intensity is 30～48.5V, frequency 0.5Hz, rat is put in self-control cellular electric shock Mus case, and connection stressor, except that the normal control group, all the other experimental group rat every morning 8:00～10:00, afternoon, 2:00～4:00 respectively accepted stress stimulation 1 time, totally 4 weeks, impose for the last time behind the stress stimulation in 2～5h with tail cover method and measure the arteria caudalis systolic pressure (after being about to rat and placing soft cotton pad to go up, the afterbody heating, to press the arteries and veins condom to the rat tails proximal part, transducer places 1/3 place, Mus tail middle and upper part, opens recording system, waits after the regular pulse appearance, utilize inflatable ball to make the cover internal pressure be elevated to the complete obiteration of beating, slowly venting then examines and reads pulse wave pairing force value from do not having to beginning to occur the time, and this is rat systolic pressure (mmHg), repeat 3 times, average).
2.2 respectively organize Hypertensive Rats and normal control group by table 2 gastric infusion every day 1 time after experimental technique becomes modeling, dosage is respectively 10ml/kg, blank group and normal control group are given normal saline, behind the successive administration 3 days, with 1% pentobarbital intraperitoneal anesthesia (5mg/kg), the carotid artery intubate is got blood, measures systolic pressure.
2.3 the experimental result experimental result sees Table following table.
The influence of table 3 pair Hypertensive Rats systolic pressure (X ± S)
| Group | Number of animals (only) | Dosage (ml/kg) | Systolic pressure before the administration | The administration after-contraction is pressed |
| Pheretima bionic enzymatic sample sets | ??10 | ??10 | ??105.36±4.63 | ??85.68±5.45 ** |
| Pheretima is the enzymolysis sample group not | ??10 | ??10 | ??104.52±2.98 | ??98.29±4.37 |
| Bombyx Batryticatus bionic enzymatic sample sets | ??10 | ??10 | ??102.11±3.61 | ??87.52±4.42 ** |
| Bombyx Batryticatus is the enzymolysis sample group not | ??10 | ??10 | ??106.46±2.34 | ??99.07±3.87 |
| The positive drug group | ??10 | ??10 | ??105.17±6.46 | ??83.86±6.05 ** |
| The blank group | ??10 | ??10 | ??107.47±5.28 | ??104.53±5.65 |
| The normal control group | ??10 | ??10 | ??78.85±4.96 | ??80.32±5.28 |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01.
Above result of the test shows that blank group and normal control group compare, and systolic pressure increases significant difference, shows the modeling success.Bionic enzymatic sample sets, positive drug group group can obviously reduce the systolic pressure of Hypertensive Rats, relatively have utmost point significant difference with the normal control group, not enzymolysis sample group and blank group relatively have notable difference but effect obviously not as the bionic enzymatic group effective.
Three, to the influence of tumor-bearing mice
1 material
1.1 animal S
180The tumor ascites mice of going down to posterity is available from the institute of oncology, Jilin Province; Kunming mouse, the SPF level, Anhui Province's Experimental Animal Center provides.
1.2 medicine is the enzymolysis group not: get Scolopendra medical material fine powder (80 order) 40g, add 10 times of amount normal saline, homogenate 30 minutes stirs evenly, and gets 1/2 amount, and microporous filter membrane (0.45 μ m) filters, promptly;
Bionic enzymatic group: get 1/2 homogenate, add 1% pepsin, regulate pH to 2.0 simultaneously, temperature is 40 ℃ ± 2 ℃, enzymolysis 4h is stirred in insulation simultaneously, transfers pH to 8.0 then, adds 1% trypsin, temperature is 50 ℃ ± 2 ℃, enzymolysis 5h is stirred in insulation simultaneously, and 85 ℃ of insulation 20min are put cold, microporous filter membrane (0.45 μ m) filters, promptly;
Other gets Scorpio medical material fine powder (80 order) 40g, prepares for test agent by the preparation method of above-mentioned Scolopendra for test agent.
Blank: normal saline.
Cyclophosphamide group: get cyclophosphamide 2g, be suspended to 100ml with water.
1.3 the reagent pepsin is available from traditional Chinese medicines group chemical reagent company limited, lot number: F20070914; Trypsin is available from traditional Chinese medicines group chemical reagent company limited, lot number: F20071228; Injection normal saline: specification: 250ml, lot number: 02110504, Zhiying Pharmaceutical Factory, Shenyang produces.
2 methods and result
2.1 test method is got Kunming mouse, is divided into 6 groups at random, 10 every group, is respectively blank group, cyclophosphamide group, bionic enzymatic sample sets, enzymolysis sample group not.Chose transplantation tumor 7 days, tumor growth is good, the tangible ascites of the abdominal tympanites mice of going down to posterity, the skin of abdomen sterilization, thrust the abdominal cavity with disposable sterilized hemostix through stomach wall, extraction ascites is put into aseptic beaker and is diluted to 1: 3 cancerous cell suspension with normal saline, in right oxter inoculation 0.2ml of every test of each group Kunming mouse.Each is organized mice and begins to irritate every day stomach next day in inoculated tumour and give corresponding test sample 0.25ml/10g, and the blank group is irritated stomach and given the equal-volume normal saline, successive administration 30 days.Administration finishes next day, puts to death after each group test mice is weighed, and separates subcutaneous lump and weighs, and respectively organizes the tumor growth situation.
2.2 the concrete experimental result of result of the test sees the following form.
Table 4 pair mice (inoculation S
180) tumor growth influence (X ± S, n=10)
Annotate: compare with matched group,
*P＜0.05,
*P＜0.01.
Result of the test shows: the bionic enzymatic sample sets has obvious antineoplastic, to S
180The tumour inhibiting rate of mice transplanted tumor is higher; And the enzymolysis sample group is not compared with matched group and is still had certain significant difference (P＜0.05), and the bionic enzymatic sample that visible the present invention makes has obvious antineoplastic.
The specific embodiment
Enumerate embodiment below, further specify the present invention, each embodiment only is used to illustrate the present invention, does not limit the present invention:
Embodiment 1
Water intaking trematodiasis 500g is ground into fine powder, adds 10 times of water gaging homogenate, regulate pH to 2.0,1% the pepsin that adds the Hirudo amount is regulated pH to 8.0 in 40 ℃ of insulation enzymolysis 2 hours, and 1% the pancreatin that adds the Hirudo amount was in 50 ℃ of insulation enzymolysis 4 hours, spray drying, add an amount of starch, granulate drying, granulate is loaded capsule.
Embodiment 2
Get Formica fusca 500g, be ground into fine powder, add 10 times of water gaging homogenate, regulate pH to 2.0,2% the pepsin that adds the Formica fusca amount is regulated pH to 8.5 in 40 ℃ of insulation enzymolysis 2 hours, and 1.5% the pancreatin that adds the Formica fusca amount was in 50 ℃ of insulation enzymolysis 5 hours, spray drying, add an amount of starch, granulate drying, granulate is loaded capsule.
Embodiment 3
Get Eupolyphaga Seu Steleophaga 500g, be ground into fine powder, add 8 times of water gaging homogenate, regulate pH to 2.6,2% the pepsin that adds the Eupolyphaga Seu Steleophaga amount is regulated pH to 7.5 in 37 ℃ of insulation enzymolysis 3 hours, and 2% the trypsin that adds the Eupolyphaga Seu Steleophaga amount was in 40 ℃ of insulation enzymolysis 6 hours, spray drying, add an amount of microcrystalline Cellulose, granulate drying, granulate, compacting are in flakes.
Embodiment 4
Water intaking Cornu Bovis seu Bubali 500g, be ground into fine powder, add 15 times of water gaging homogenate, regulate pH to 3.0,5% the pepsin that adds the Cornu Bubali amount was in 40 ℃ of insulation enzymolysis 2 hours, regulate pH to 8.5,5% the trypsin that adds the Cornu Bubali amount is heated to 85 ℃ of insulations 15 minutes in 45 ℃ of insulation enzymolysis 5 hours, filter, filtrate is divided and is got the following solution of molecular cut off 5kD with the ultrafilter membrane ultrafiltration, makes injection.
Embodiment 5
Get Pheretima 500g, be ground into fine powder, add 15 times of water gaging homogenate, regulate pH to 1.5,0.5% the pepsin that adds the Pheretima amount is regulated pH to 8.0 in 40 ℃ of insulation enzymolysis 1 hour, 1.5% the trypsin that adds the Pheretima amount was in 50 ℃ of insulation enzymolysis 3 hours, be heated to 95 ℃ of insulations 30 minutes, filter, filtrate is with the ultrafilter membrane ultrafiltration, divide and get the following solution of molecular cut off 5kD, add mannitol, lyophilized injectable powder is made in lyophilization.
Embodiment 6
Get Bombyx Batryticatus 500g, be ground into fine powder, add 15 times of water gaging homogenate, regulate pH to 2.5,4% the pepsin that adds the Bombyx Batryticatus amount is regulated pH to 8.5 in 40 ℃ of insulation enzymolysis 2 hours, and 5% the trypsin that adds the Bombyx Batryticatus amount was in 45 ℃ of insulation enzymolysis 6 hours, spray drying, add an amount of starch, granulate drying, granulate is loaded capsule.
Embodiment 7
Get Periostracum Cicadae 500g, be ground into fine powder, add 10 times of water gaging homogenate, regulate pH to 2.0,2% the pepsin that adds the Periostracum Cicadae amount was in 40 ℃ of insulation enzymolysis 2.5 hours, regulate pH to 8.0,2% the pancreatin that adds the Periostracum Cicadae amount is heated to 85 ℃ of insulations 15 minutes in 50 ℃ of insulation enzymolysis 5 hours, filter, filtrate is divided and is got the following solution of molecular cut off 5kD with the ultrafilter membrane ultrafiltration, makes spray.
Embodiment 8
Get Scorpio 500g, be ground into fine powder, add 12 times of water gaging homogenate, regulate pH to 2.5,2.5% the pepsin that adds the Scorpio amount is regulated pH to 8.0 in 39 ℃ of insulation enzymolysis 2.5 hours, and 3% the trypsin that adds the Scorpio amount was in 40 ℃ of insulation enzymolysis 6 hours, spray drying, add an amount of microcrystalline Cellulose, granulate drying, granulate, compacting are in flakes.
Embodiment 9
Get Scolopendra 500g, be ground into fine powder, add 15 times of water gaging homogenate, regulate pH to 2.0,2.5% the pepsin that adds the Scolopendra amount is regulated pH to 8.5 in 40 ℃ of insulation enzymolysis 3 hours, and 2.5% the pancreatin that adds the Scolopendra amount was in 50 ℃ of insulation enzymolysis 6 hours, spray drying, add an amount of Icing Sugar and dextrin, granulate drying, granulate is made granule.
Embodiment 10
Get Pheretima 500g, be ground into fine powder, add 15 times of water gaging homogenate, regulate pH to 2.0,2.5% the pepsin that adds the Pheretima amount is regulated pH to 8.5 in 40 ℃ of insulation enzymolysis 3 hours, and 2.5% the pancreatin that adds the Pheretima amount was in 50 ℃ of insulation enzymolysis 6 hours, spray drying, add an amount of Icing Sugar and dextrin, granulate drying, granulate is made granule.
Embodiment 11
Get Scorpio 250g, Bombyx Batryticatus 100g, add 15 times of water gaging homogenate, regulate pH to 2.0,1% the pepsin that adds total dose is regulated pH to 8.0 in 40 ℃ of insulation enzymolysis 2 hours, and 1% the pancreatin that adds total dose was in 50 ℃ of insulation enzymolysis 4 hours, be heated to 85 ℃ of insulations 15 minutes, filter, filtrate is with the ultrafilter membrane ultrafiltration, divide and get the following solution of molecular cut off 10kD, add carbomer, swollen 12 hours, transfer PH to increase viscosity, add the 150g Rhizoma Gastrodiae and extract concentrated solution, mixing is made gel.
Embodiment 12
Get the observation of curative effect of embodiment 6 made capsule for treating hypertension.
This organizes 60 examples, wherein male 32 examples, women 28 examples; Year mean age (56.5 soil 7.9);
The course of disease: the course of disease (9.20 scholar 5.90) year; 1 grade of 18 example of hypertension, 2 grade of 34 example, 3 grade of 8 example.
Diagnostic criteria: hypertension is meant under the situation of not taking antihypertensive drug, systolic pressure 〉=18.62kPa (140mmHg) and (or) diastolic pressure 〉=11.79kPa (90mmHg); The diagnosis of EH should record blood pressure at least 2 times and reach hypertensive diagnostic criteria under non-quiescent condition on the same day, and after getting rid of secondary hypertension, can make a definite diagnosis.
Therapeutic Method: oral administration, every day, dose was equivalent to 15g crude drug amount; Serve on 4 all observe the curative effect.
Efficacy of antihypertensive treatment evaluation criteria: 1. produce effects: diastolic pressure descends and is not less than 1.33kPa (10mmHg), and reaches normal range; Descended though diastolic pressure is not reduced to normally and to be not less than 2.66kPa (20mmHg); 2. effective: diastolic pressure descends less than 1.33kPa (10mmHg), but has reached normal range; Decline 1.33～2.53kPa before diastolic pressure is treated (10～19mmHg), but do not reach normal range; Systolic pressure preceding decline of treatment is not less than 4.0kPa (30mmHg); 3. invalid: as not reach effective standard.
Therapeutic outcome: this is organized in 60 examples, clinical produce effects 24 examples (40%), and effective 33 examples (55%), invalid 3 examples (5%), total effective rate is 95%.
Claims (10)
1, a kind of bionic enzymatic hydrolysate of animal drugs is characterized in that this product prepares by the following method:
Get animal drugs, be ground into fine powder, add water, regulate pH to 1.0～3.0, add pepsin in 35～45 ℃ of insulation enzymolysis 0.5～4.0 hour, re-adjustment pH to 7.5～8.5 add pancreatin or trypsin in 40～50 ℃ of insulation enzymolysis 2.0～8.0 hours, promptly.
2, the bionic enzymatic hydrolysate of animal drugs according to claim 1 is characterized in that this product prepares by the following method:
Get animal drugs, be ground into fine powder, add water homogenate, regulate pH to 1.5～2.5,0.5%～5.0% the pepsin that adds the animal dose was in 35～45 ℃ of insulation enzymolysis 1.0～3.0 hours, re-adjustment pH to 7.5～8.5 add 0.5%～5.0% pancreatin of animal dose or trypsin in 40～50 ℃ of insulation enzymolysis 3.0～6.0 hours, promptly.
3, the enzymatic hydrolysate of animal drugs according to claim 2 is characterized in that this product prepares by the following method:
Get animal drugs, be ground into fine powder, add water homogenate, regulate pH to 2.0,1.0%～2.0% the pepsin that adds the animal dose was in 40 ℃ of insulation enzymolysis 1.0～3.0 hours, re-adjustment pH to 8.0 adds 1.0%～2.0% pancreatin of animal dose or trypsin in 50 ℃ of insulation enzymolysis 3.0～6.0 hours, promptly.
4,, it is characterized in that animal drugs is ground into fine powder to be added and be heated to 80～100 ℃ and be incubated 15～30 minutes after the water homogenate earlier, is cooled to the temperature required enzymolysis that carries out again according to the enzymatic hydrolysate of arbitrary described animal drugs in the claim 1 to 3.
5, according to the enzymatic hydrolysate of arbitrary described animal drugs in the claim 1 to 3, it is characterized in that used pepsic enzyme activity is not less than 1200U/g, tryptic enzyme activity is not less than 2500U/g, and the casein conversion power of pancreatin is not less than 25.0.
6, the animal drugs in the enzymatic hydrolysate of arbitrary described animal drugs can be Cornu Bubali, Hirudo, Tabanus, Formica fusca, Eupolyphaga Seu Steleophaga, Scolopendra, Scorpio, Bombyx Batryticatus, Periostracum Cicadae, Pheretima in the claim 1 to 5.
7,, it is characterized in that adding conventional pharmaceutic adjuvant and make injection, oral formulations or external preparation according to the enzymatic hydrolysate of arbitrary described animal drugs in the claim 1 to 5.
8, the enzymatic hydrolysate of arbitrary described animal drugs is used for the application of the medicine of hepatitis virus resisting in the claim 1 to 5 in preparation.
9, the enzymatic hydrolysate of arbitrary described animal drugs is used for the application of the medicine of blood pressure lowering in the claim 1 to 5 in preparation.
10, the enzymatic hydrolysate of arbitrary described animal drugs is used for the treatment of application in the medicine of cancer in preparation in the claim 1 to 5.
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