CN105535924A - Compound lumbricus small peptide composition for treating cardiovascular diseases - Google Patents
Compound lumbricus small peptide composition for treating cardiovascular diseases Download PDFInfo
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Abstract
The invention discloses a compound lumbricus small peptide effective part composition for treating cardiovascular diseases, and belongs to the field of traditional Chinese medicine. The composition is prepared by mixing lumbricus small peptide extracted from lumbricus, uncaria total alkaloid extracted from uncaria, ligusticum wallichii total alkaloids extracted from ligusticum wallichii and milkvetch root total saponins extracted from milkvetch roots. Pharmacy conventional accessories are added, various dosage forms are formed, and the compound lumbricus small peptide effective part composition can be clinically used for treating hypertension, hyperviscosity, coronary disease, stenocardia and other diseases.
Description
Technical field
The present invention relates to and a kind ofly have Chinese medicinal effective-part composition of Cardiovarscular effect and preparation method thereof, Chinese medicinal effective-part composition specifically extracted from Pheretima, Ramulus Uncariae Cum Uncis, Rhizoma Chuanxiong, the Radix Astragali and preparation method thereof, belongs to the field of Chinese medicines.
Background technology
Cardiovascular disease, being also called blood circulation diseases, is a series of diseases relating to blood circulation, and current cardiovascular disease has become worldwide one " modern popular is sick ", the trend that its sickness rate and case fatality rate raise year by year is day by day obvious, and production and the life of the mankind in serious threat.Often seem comparatively difficulty in the process of Cardiovarscular.In order to cardiovascular disease can be controlled, various anti-cardiovascular disease must be taken to patient.Because Drug therapy can only control cardiovascular disease in a certain degree, Cardiovarscular can only lean on medicine to maintain for a long time.Once drug failure or drug withdrawal, cardiovascular disease may occur again, and the level of preventing and treating therefore improving cardiovascular disease is imperative.
At present, doctor trained in Western medicine mainly adopts the medicines such as Statins, beta-blocker class and angiotensin receptor antagonist to the treatment of cardiovascular disease, although Western medicine achieves very large progress decades in the past, but Western medicine mostly is the medicine of single target spot or a few target spot, the chronic disease being used for treating this complexity for a long time can bring serious side effect.In addition, cardiovascular disease is a kind of complex disease, there is complementation and easily occur body Drug resistance between path, and the easy producer sudden change of long-term prescription also easily forms the Drug resistance of body.
The traditional Chinese medical science is thought, cardiovascular disease belongs to the category of the traditional Chinese medical science " cardiopalmus ", " strongly fragrant disease ", is hindered cause the five internal organs mechanism of qi to get along well to be formed by depression of liver-QI, feelings will, main with liver, spleen, the heart etc. internal organs get involved relevant with disorder of QI and blood.Or due to disgruntled smooth, cause that liver is lacked of proper care, gas loses catharsis, make the formation that depression of liver-QI causes blood tired, and stagnation of liver-QI can cause and accumulates humidogene stagnation of phlegm network, therefore vexed easily dry, palpitation and uneasiness.Or cause deficiency of both the heart and spleen because feelings will is unsuccessful, consume sad gas, insufficiency of vital energy and blood and stagnation of liver-QI and press down spleen etc., cause the timid and vexed kind worry of insomnia and dreamful sleep, cardiopalmus etc.Due to the stagnation of QI, cardiovascular disease occurs that phlegm-damp blood is tired at the initial stage of a disease, be mainly excess syndrome, should based on dampness removing promoting the circulation of blood, activating QI to eliminate phlegm, tranquilizing mind treatment according to the state of an illness.Sthenia is transforming into asthenia for the disease of patients on long-term, as dialectical for strongly fragrantly for a long time to overtax one's nerves, hyperactivity of fire caused by deficiency of YIN and heart spleen all lose, should to calm the nerves treatment based on blood yiqi, righting.And from China's Chinese traditional herbs (Chinese medicine compound or single medicinal material), extract the important component part that the cardiovascular disease medicine that obtains medical treatment has become the research of Present Domestic TCM Modernization, resources of Chinese medicinal herb is extensive, various in style, for the development of new Cardiovarscular medicine provides good basis.
A large amount of protease and peptidase is there is in animal body, past, people thought always, after plant animal protein is degraded into little peptide, then is degraded to free amino acid and could be absorbed by animal, thus people always using aminoacid as the Major Nutrient studied, make, pursue, take.In the sixties in 20th century, scientist just produces convincing evidence proves that little peptide can by absorbed intact and utilization.Due to the discovery that this is great, people just progressively receive the viewpoint that peptide directly can be absorbed by animal.Little peptide and amino acid whose mixture the absorbance of human body and infiltration rate all good than simple aminoacid, moreover, little peptide can also protect easily destroyed aminoacid in intestinal.In addition, simple aminoacid newly forms peptides and proteins in vivo as required, and little peptide and amino acid whose mixture can save the process that a part reconfigures, and therefore, its biological value is higher.
Prepared slices of Chinese crude drugs active constituent content is low, and dose is large, inconvenient, and flavour of a drug are bitter, not easily accepted by patient, but through the extraction of modern science technique, active constituent content is improved, consumption reduces, taking convenience, and good medicine is no longer bitter to the taste, adds the crowd of acceptance.
Summary of the invention
One object of the present invention is to provide a kind of Chinese medicinal effective-part composition with Cardiovarscular effect;
Another object of the present invention is the preparation method and the purposes that provide above-mentioned Chinese medicine composition.
The first object of the present invention: with the little peptide of Pheretima, Rhizoma Chuanxiong total alkaloids, uncaria total alkaloids, Radix Astragali total saponins for raw material, add proper pharmaceutical excipients, make relevant dosage form.
The composition of the crude drug of the compositions of medicine of the present invention and proportioning are: Pheretima little peptide 6 weight portion, uncaria total alkaloids 8 weight portion, Rhizoma Chuanxiong total alkaloids 3 weight portion, Radix Astragali total saponins 6 weight portion.
In above-mentioned total alkaloids effective site, the content of total alkaloids is not less than 50% of extract, and in total saponins effective site, total saponins is not less than 60% of extract, and in little peptide effective site, the content of the little peptide of Pheretima is not less than 50% of extract
This pharmaceutical composition can add corresponding adjuvant, makes any one in tablet, granule, hard capsule.Then can add other effective site with Cardiovarscular medicine, effective ingredient, synthetic or semi-synthetics again, and then add corresponding adjuvant, make any one in tablet, granule, hard capsule.
The preparation method of Chinese medicine composition of the present invention comprises the following steps:
1. the preparation of the little peptide of Pheretima: get Pheretima, suitable pulverizing, add 10 times of water gaging homogenate 15min, heated and boiled 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0, then the pepsin of amount of substrate 5% is added, enzymolysis 3h under 37 DEG C of conditions, then sodium hydroxide solution is adopted to regulate pH to 8, add the trypsin of amount of substrate 5%, enzymolysis 6h under 45 DEG C of conditions, stir constantly and regulate pH 8, continue intensification after enzymolysis and boil 15min, let cool, centrifugal, collect supernatant, regulate pH7.0, add the active carbon of 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Hirudo, wherein the content of the little peptide of Pheretima is 50% ~ 90%,
2. the preparation of uncaria total alkaloids: get Uncaria, add medical material weight 10 times of 75% ethanol 75 DEG C of reflux, extract, 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol is 1:2 to relative density, adds heavy 1% hydrochloric acid 20% alcoholic solution such as medical material weight, with the flow velocity of 1ml/min by 732 type strongly acidic styrene type cation exchange resins, 8BV is washed to neutrality, 80% ethanol 6BV remove impurity, 10BV1% ammonia 80% ethanol elution, eluant decompression and solvent recovery, drying, obtains uncaria total alkaloids; Wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%;
3. the preparation of Rhizoma Chuanxiong total alkaloids: get Ligusticum chuanxiong Hort, adds medical material weight 8 times amount 90% ethanol 70 DEG C of reflux, extract, 2 times, each 2 hours, and merging filtrate merges reclaim under reduced pressure, and dry, obtain Rhizoma Chuanxiong total alkaloids, wherein the content of ligustrazine is 0.23% ~ 0.24%;
4. the preparation of Radix Astragali total saponins: get Milkvetch Root, add 70% ethanol, 80 DEG C of reflux, extract, 1 hour of medical material weight 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1, with the flow velocity of 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 times of column volumes, again with 70% ethanol elution of 4 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
5. get above-mentioned four parts of effective sites according to the weight proportion of each effective site, do not add adjuvant or add pharmaceutics customary adjuvant, make pharmaceutically acceptable dosage form.
The content assaying method of uncaria total alkaloids is: the drafting of standard curve: it is appropriate that precision takes reference substance, adds 2% hydrochloric acid solution and makes solution containing Ramulus Uncariae cum Uncis alkali 100ug/ml, product solution in contrast.Accurate draw reference substance solution 0.2,0.4,0.6,0.8,1.0ml is placed in 10ml measuring bottle respectively, is diluted to scale with 2% hydrochloric acid solution.With corresponding solvent for blank, measure trap at 250nm wavelength place, take trap as vertical coordinate, concentration is abscissa, drawing standard curve.Get uncaria total alkaloids effective site 25mg, be placed in 50ml measuring bottle, add 2% dissolving with hydrochloric acid and be diluted to scale.Accurate absorption 2ml, puts in 100ml measuring bottle, is diluted to scale with 2% hydrochloric acid solution, with corresponding solvent for blank, measures trap, calculate content, to obtain final product by standard curve at 250nm wavelength place.
Rhizoma Chuanxiong total alkaloids content assaying method is: the drafting of standard curve: take ligustrazine hydrochloride reference substance 25mg, accurately weighed, puts in 25ml measuring bottle, dissolves and be diluted to scale, shake up with chloroform.Obtain the reference substance solution of 1mg/ml. accurate draw ligustrazine hydrochloride reference substance solution 0,2,3,4,5,6,7,8ml is placed in separatory funnel, add 7ml bromothymol blue solution, shake well, colour developing layering, collect chloroform layer, add chloroform extraction to chloroform layer is colourless.Combined chloroform layer, is settled to 10ml, shakes up, and filters.Blank solution is obtained with legal system.Absorbance is measured in 415nm place.Take trap as vertical coordinate, concentration is abscissa, drawing standard curve.Get Rhizoma Chuanxiong total alkaloids effective site 20mg, the hydrochloric acid 60ml adding 1mol/L dissolves, filter, after filtrate adjusts pH9 ~ 10 with strong aqua ammonia, use chloroform extraction again 3 times (40,30,20ml), combined chloroform extracting solution, the methanol hydrochloride solution adding 1mol/L adjusts pH1.2, puts evaporate to dryness in 45 DEG C of waters bath with thermostatic control.Residue adds chloroform and dissolves, and is settled to 10ml, shakes up, obtain need testing solution.Precision pipettes need testing solution 4ml in separatory funnel, adds 7ml bromothymol blue solution, shake well, and colour developing layering, collects chloroform layer, adds chloroform extraction to chloroform layer is colourless.Combined chloroform layer, is settled to 10ml, shakes up, and filters, obtains blank solution with legal system, measures absorbance in 415nm, and according to standard curve Equation for Calculating content.
Radix Astragali total saponins content assaying method is: astragaloside reference substance is appropriate, adds dissolve with methanol and is quantitatively diluted to the solution of every 1ml containing 0.5mg, to obtain final product.Precision measure reference substance solution 0.2,0.4,0.6,0.8,1.0ml is placed in 10ml respectively and puts tool plug test tube, water-bath volatilizes solvent, and precision adds 5% vanillin glacial acetic acid solution 1ml and perchloric acid 4ml, mixing, puts in 60 DEG C of water-baths and heats 15min, use water cooling immediately, precision adds glacial acetic acid 15ml, shaking up, is blank with corresponding reagent, according to ultraviolet visible spectrophotometry, absorbance is measured at 530nm wavelength place, take trap as vertical coordinate, concentration is abscissa, drawing standard curve.Get Radix Astragali total saponins effective site 5mg residue dissolve with methanol and be transferred in 10ml measuring bottle, adding methanol dilution to scale, shake up, precision measures subsequent filtrate 1ml, puts in 10ml measuring bottle, adds methanol dilution to scale, shakes up, to obtain final product.Precision measures need testing solution 1ml, puts in tool plug test tube, water-bath volatilizes solvent, precision adds 5% vanillin glacial acetic acid solution 1ml and perchloric acid 4ml, and mixing, puts in 60 DEG C of water-baths and heat 15min, use water cooling immediately, precision adds glacial acetic acid 15ml, shakes up, and take corresponding reagent as blank, according to ultraviolet visible spectrophotometry, measure absorbance at 530nm wavelength place, substitute in standard curve and calculate, to obtain final product.
After the contrast preferably of inventor, the optimum weight proportioning determining above-mentioned effective site is: Pheretima little peptide 6 weight portion, uncaria total alkaloids 8 weight portion, Rhizoma Chuanxiong total alkaloids 3 weight portion, Radix Astragali total saponins 6 weight portion.Proportionally get the little peptide of above-mentioned Pheretima, uncaria total alkaloids, Rhizoma Chuanxiong total alkaloids, Radix Astragali total saponins, do not add adjuvant or add pharmaceutics customary adjuvant, make pharmaceutically acceptable dosage form.
The dosage form that pharmaceutical composition of the present invention can be made into is tablet, granule, hard capsule.
Said pharmaceutics customary adjuvant includes but not limited to binding agent above, as cellulose derivative, alginate, gelatin and polyvinylpyrrolidone; Diluent, starch, Icing Sugar, dextrin, lactose, microcrystalline Cellulose, mannitol; Lubricant, as Pulvis Talci, magnesium stearate, micropowder silica gel and Polyethylene Glycol; Disintegrating agent, as carboxymethylstach sodium, low-substituted hydroxypropyl methylcellulose, cross-linked carboxymethyl cellulose; Absorption enhancer, as quaternary ammonium compound; Surfactant, as liquor-saturated in hexadecane, sodium lauryl sulphate; Excipient, as lactose, starch, sucrose, dextrin, calcium carbonate, aluminum sulfate, magnesium oxide, magnesium stearate, sodium bicarbonate, glycerol, propylene glycol, Polyethylene Glycol, Sorbitol, lanoline, vaseline, microcrystalline Cellulose, Cera Flava, wood cured, liquid paraffin, resin, advanced wax, pressure sensitive adhesive, semi-synthetic fatty acid ester etc.
Active component composition of the present invention is used for the treatment of cardiovascular disease.
Beneficial effect
Chinese medicine composition of the present invention, is form in conjunction with modern medicine study achievement prescription by traditional Chinese medicine theory simultaneously, possesses Chinese medicine characteristic, meet the Therapeutic Principle of modern medicine simultaneously.
Modern life rhythm is nervous, and the increasing pressure of family, cause is large, and the emotion of people is also more and more unstable; Simultaneously, excessive consumption of alcohol, the motion taken in too many food fat, lack necessity, in addition the pollution of living environment, these factors directly cause human metabolism's speed to slow down, velocity of blood flow can slow down, and blood viscosity raises rapidly, causes cerebral ischemia, if prevent not in time, nurse one's health, the cardiovascular and cerebrovascular diseases such as coronary heart disease, hypertension, cerebral thrombosis, fatty liver will be caused.Long-term hypertension can make cerebral artery vessel wall thickening or hardening, and tube chamber attenuates.When blood pressure rises sharply, cerebrovascular easily breaks generation cerebral hemorrhage; Or the brain small artery hardened forms a kind of microaneurysm of chestnut grain size, when fluctuations in blood, arteriole stream breaks and causes cerebral hemorrhage; Or hypertension accelerates arteriosclerosis process, arterial endothelial cell liquid sustains damage, and platelet is easily assembled in injury, easily forms brain blood pressure bolt again, causes cardiovascular and cerebrovascular disease.
The present invention take Traditional Chinese medical theory as guiding theory, simultaneously in conjunction with correlational study achievement both domestic and external, and Be very effective in Cardiovarscular.Take Pheretima as monarch drug in side, Pheretima is salty, cold, returns liver, spleen, urinary bladder channel.Function heat clearing away is calmed the frightened, and dredging collateral, relievings asthma, diuresis, and for cerebral thrombosis, cerebral infarction, coronary heart disease, apoplexy, joint is benumbed, numb limbs and tense tendons, hemiplegia, the diseases such as vascular hypertension.Therefore be monarch; The Radix Astragali is ministerial drug, and the Radix Astragali is sweet, tepor.Return lung, spleen channel.Have tonifying Qi and lifting yang, strengthening superficial resistance to stop perspiration, inducing diuresis to remove edema, promote the production of body fluid and nourish blood, effect of the stagnant blood stasis dispelling of row, modern medicine study shows, modern study shows, no matter merely the effective ingredient astragalus polysaccharides, Radix Astragali saponin etc. of the Radix Astragali or the Radix Astragali all have the Cardiovascular of affirmative.It, by suppressing the activity that Na+-K+-ATPase is active, increase myocardium beta receptor number, suppress phosphodiesterase (PDE), plays dual regulation effect to Isolated Toad Heart.In addition, the Radix Astragali can also affect blood pressure, reduces blood viscosity and improve abnormal hemorheology index.Ramulus Uncariae Cum Uncis is adjuvant drug, and Ramulus Uncariae Cum Uncis is sweet, cool.Return liver, pericardium channel.Have dispelling wind and relieving convulsion, effect of heat clearing away suppressing the hyperactive liver, dizzy for having a headache, dysphoric cold, infantile convulsion are twitched, pre-eclampsia, the diseases such as hypertension.Be one of first-selected medical material for the treatment of cardiovascular and cerebrovascular disease, have long applicating history clinically.Rhizoma Chuanxiong, for making medicine, is usually used in blood-activating and qi-promoting, wind-expelling pain-stopping, and the pungent temperature of Rhizoma Chuanxiong is fragrant dry, walks and does not keep, can go loose, and the up mountain peak that reaches is pushed up; Enter blood system again, descendingly reach the sea of blood.Blood circulation promoting and blood stasis dispelling effect is extensive, the suitable various disease of stagnation of blood stasis.In we, each medicine complements each other, indispensable, by traditional Chinese medical science monarch, minister, helps, makes theoretical reasonable formula, play the merit of dampness removing promoting the circulation of blood, activating QI to eliminate phlegm, blood pressure lowering altogether.
Product composition of the present invention is clear and definite, stable curative effect, dose is little, safety is high, not only meet the principles of formulating prescriptions of Chinese Traditional Medicine Cardiovarscular, and meet the demand for development of the modernization of Chinese medicine, promotion medicinal industry to be changed and significant to promotion China internationalization of tcm from copying to autonomous innovation.Through consulting pertinent literature and patent, do not retrieve with the little peptide of Pheretima, Radix Astragali total saponins, uncaria total alkaloids, Rhizoma Chuanxiong total alkaloids as raw material, production and processing also makes the treatment of Suitable pharmaceutical preparations for cardiovascular disease or the report of auxiliary treatment.
For verifying pharmacological action of the present invention further, carry out animal pharmacodynamic experiment with Tablets.
Content of the test: to the hypotensive effect of salt density value and salt hypertension of resistance rat model.
1 reagent, sample and animal
1.1 reagent
Edible salt
1.2 test sample
Sample 1:(Tablets)
Get Pheretima 10kg, suitable pulverizing, add 10 times of water gaging homogenate 15min, heated and boiled 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0, then the pepsin of amount of substrate 5% is added, enzymolysis 3h under 37 DEG C of conditions, then sodium hydroxide solution is adopted to regulate pH to 8, add the trypsin of amount of substrate 5%, enzymolysis 6h under 45 DEG C of conditions, stir constantly and regulate pH 8, continue intensification after enzymolysis and boil 15min, let cool, centrifugal, collect supernatant, regulate pH7.0, add the active carbon of 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Hirudo, wherein the content of the little peptide of Pheretima is 50% ~ 90%, get Uncaria 10kg, add medical material weight 10 times of 75% ethanol 75 DEG C of reflux, extract, 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol is 1:2 to relative density, adds heavy 1% hydrochloric acid 20% alcoholic solution such as medical material weight, with the flow velocity of 1ml/min by the 732 type strongly acidic styrene type cation exchange resins identical with medical material weight, 8BV is washed to neutrality, 80% ethanol 6BV remove impurity, 10BV1% ammonia 80% ethanol elution, eluant decompression and solvent recovery, drying, obtains uncaria total alkaloids, wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%, get Ligusticum chuanxiong Hort 10kg, add medical material weight 8 times amount 90% ethanol 70 DEG C of reflux, extract, 2 times, each 2 hours, merging filtrate merges reclaim under reduced pressure, and dry, obtain Rhizoma Chuanxiong total alkaloids, wherein the content of ligustrazine is 0.23% ~ 0.24%, get Milkvetch Root 10kg, add 70% ethanol, 75 DEG C of reflux, extract, 1 hour of medical material weight 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1, with the flow velocity of 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 times of column volumes, again with 70% ethanol elution of 4 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
Get the little peptide 60g of Pheretima, uncaria total alkaloids 80g, Rhizoma Chuanxiong total alkaloids 30g, Radix Astragali total saponins 60g, add hypromellose 14g, microcrystalline Cellulose 5g, micropowder silica gel 1g adds lactose again and make tablet to 300g, every sheet 0.6g.
Sample 2:(capsule of the present invention)
Adopt effective site raw material prepared by sample 1.
Get the little peptide 60g of Pheretima, uncaria total alkaloids 80g, Rhizoma Chuanxiong total alkaloids 30g, Radix Astragali total saponins 60g, add starch to 500g, load No. 0 capsule, the tolerant 0.5g of every intragranular, makes capsule.
Sample 3:
Adopt effective site raw material prepared by sample 1.
Get the little peptide 30g of Pheretima, uncaria total alkaloids 90g, Rhizoma Chuanxiong total alkaloids 10g, Radix Astragali total saponins 100g, add starch to 500g, load No. 0 capsule, the tolerant 0.5g of every intragranular, makes capsule.
Sample 4:
Adopt effective site raw material prepared by sample 1.
Get uncaria total alkaloids 80g, Rhizoma Chuanxiong total alkaloids 30g, Radix Astragali total saponins 60g, add starch to 500g, load No. 0 capsule, the tolerant 0.5g of every intragranular, makes capsule.
Before administration, get above-mentioned corresponding preparations, porphyrize, adopt pure water to be mixed with required drug level.
1.3 laboratory animal
Healthy male mice 40, is provided by Shandong Traditional Chinese Medicine University's experimental animal center.
2 experimental techniques
Healthy male rat 40 is divided into 6 groups at random: blank group, model group, 1,2,3,4 groups, sample.With reference to modeling method, with high salt diet (8% sodium chloride) and Lithyronine process rat, a part of rat blood pressure raises, and the blood pressure of another part animal is without significant change.Rat high and minimum for blood pressure is carried out inbreeding respectively, San Daihou, isolate salt density value rat.Setting normal group; Equal-volume normal saline group; The groups such as the present invention's (1. 2. 3. 4.) group.Dosage is 5mg.kg
-1.Continue medication to the 8th day, normal group and model group give equal-volume distilled water and normal saline respectively simultaneously.Mice is placed in metabolic cage normally raise, measures arteria caudalis blood pressure every day 2 times.Blood pressure level is according to documentation standards scoring (0 point: the normal 85 ~ 110mmHg of blood pressure; 1 point: mild hypertension, blood pressure 110mmHg ~ 120mmHg; 2 points: moderate hypertension, pressure value 120mmHg ~ 130mmHg; 3 points: severe hypertension, more than blood pressure 130mmHg.
Of the present invention group (1. 2.) has good hypotensive effect and matched group (p<0.05) to compare with model group (p<0.05) for the Hypertension Rats caused by salt density value and all has the significance difference opposite sex.The results are shown in Table 1.
The result of the test of Hypertension Rats caused by table 1 pair salt density value
Note: compare P < 0.001 with normal group; P < 0.05, P < 0.01 is compared with model group.
Illustrate that of the present invention group (1. 2.) has therapeutical effect to Hypertension Rats caused by salt density value.
Detailed description of the invention
Illustrate technical scheme of the present invention further below by way of specific embodiment, but the claimed content of the present patent application not only that.Consumption in actual production is not limited only to the drug dose in example, can use the measurement unit such as ton, kilogram, gram, but the consumption proportion of each material medicine is still according to the technical scheme that we are bright.
Embodiment 1:
Prescription: the little peptide 60g of Pheretima, uncaria total alkaloids 80g, Rhizoma Chuanxiong total alkaloids 30g, Radix Astragali total saponins 60g,
Method for making: the preparation of the little peptide of Pheretima: get Pheretima, suitable pulverizing, add 10 times of water gaging homogenate 15min, heated and boiled 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0, then the pepsin of amount of substrate 5% is added, enzymolysis 3h under 37 DEG C of conditions, then sodium hydroxide solution is adopted to regulate pH to 8, add the trypsin of amount of substrate 5%, enzymolysis 6h under 45 DEG C of conditions, stir constantly and regulate pH 8, continue intensification after enzymolysis and boil 15min, let cool, centrifugal, collect supernatant, regulate pH7.0, add the active carbon of 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Hirudo, wherein the content of the little peptide of Pheretima is 50% ~ 90%,
The preparation of uncaria total alkaloids: get Uncaria, add medical material weight 10 times of 75% ethanol 75 DEG C of reflux, extract, 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol is 1:2 to relative density, adds heavy 1% hydrochloric acid 20% alcoholic solution such as medical material weight, with the flow velocity of 1ml/min by 732 type strongly acidic styrene type cation exchange resins, 8BV is washed to neutrality, 80% ethanol 6BV remove impurity, 10BV1% ammonia 80% ethanol elution, eluant decompression and solvent recovery, drying, obtains uncaria total alkaloids; Wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%;
The preparation of Rhizoma Chuanxiong total alkaloids: get Ligusticum chuanxiong Hort, adds medical material weight 8 times amount 90% ethanol 70 DEG C of reflux, extract, 2 times, each 2 hours, and merging filtrate merges reclaim under reduced pressure, and dry, obtain Rhizoma Chuanxiong total alkaloids, wherein the content of ligustrazine is 0.23% ~ 0.24%;
The preparation of Radix Astragali total saponins: get Milkvetch Root, add 70% ethanol, 80 DEG C of reflux, extract, 1 hour of medical material weight 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1, with the flow velocity of 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 times of column volumes, again with 70% ethanol elution of 4 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
Get above-mentioned each effective site, pulverize, mixing, adds starch to 500g, and mixing is granulated, incapsulated, make 1000, to obtain final product.
Embodiment 2:
Prescription: the little peptide 120g of Pheretima, uncaria total alkaloids 160g, Rhizoma Chuanxiong total alkaloids 60g, Radix Astragali total saponins 120g
Method for making: the preparation of the little peptide of Pheretima: get Pheretima, suitable pulverizing, add 10 times of water gaging homogenate 15min, heated and boiled 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0, then the pepsin of amount of substrate 5% is added, enzymolysis 3h under 37 DEG C of conditions, then sodium hydroxide solution is adopted to regulate pH to 8, add the trypsin of amount of substrate 5%, enzymolysis 6h under 45 DEG C of conditions, stir constantly and regulate pH 8, continue intensification after enzymolysis and boil 15min, let cool, centrifugal, collect supernatant, regulate pH7.0, add the active carbon of 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Hirudo, wherein the content of the little peptide of Pheretima is 50% ~ 90%,
The preparation of uncaria total alkaloids: get Uncaria, add medical material weight 10 times of 75% ethanol 75 DEG C of reflux, extract, 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol is 1:2 to relative density, adds heavy 1% hydrochloric acid 20% alcoholic solution such as medical material weight, with the flow velocity of 1ml/min by 732 type strongly acidic styrene type cation exchange resins, 8BV is washed to neutrality, 80% ethanol 6BV remove impurity, 10BV1% ammonia 80% ethanol elution, eluant decompression and solvent recovery, drying, obtains uncaria total alkaloids; Wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%;
The preparation of Rhizoma Chuanxiong total alkaloids: get Ligusticum chuanxiong Hort, adds medical material weight 8 times amount 90% ethanol 70 DEG C of reflux, extract, 2 times, each 2 hours, and merging filtrate merges reclaim under reduced pressure, and dry, obtain Rhizoma Chuanxiong total alkaloids, wherein the content of ligustrazine is 0.23% ~ 0.24%;
The preparation of Radix Astragali total saponins: get Milkvetch Root, add 70% ethanol, 80 DEG C of reflux, extract, 1 hour of medical material weight 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1, with the flow velocity of 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 times of column volumes, again with 70% ethanol elution of 4 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
Get above-mentioned each effective site, pulverize, mixing, add hypromellose 28g, microcrystalline Cellulose 10g, micropowder silica gel 2g adds lactose again to 300g, mixing, is pressed into tablet, makes 1000, to obtain final product.
Embodiment 3:
Prescription: the little peptide 600g of Pheretima, uncaria total alkaloids 800g, Rhizoma Chuanxiong total alkaloids 300g, Radix Astragali total saponins 600g
Method for making: the preparation of the little peptide of Pheretima: get Pheretima, suitable pulverizing, add 10 times of water gaging homogenate 15min, heated and boiled 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0, then the pepsin of amount of substrate 5% is added, enzymolysis 3h under 37 DEG C of conditions, then sodium hydroxide solution is adopted to regulate pH to 8, add the trypsin of amount of substrate 5%, enzymolysis 6h under 45 DEG C of conditions, stir constantly and regulate pH 8, continue intensification after enzymolysis and boil 15min, let cool, centrifugal, collect supernatant, regulate pH7.0, add the active carbon of 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Hirudo, wherein the content of the little peptide of Pheretima is 50% ~ 90%,
The preparation of uncaria total alkaloids: get Uncaria, add medical material weight 10 times of 75% ethanol 75 DEG C of reflux, extract, 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol is 1:2 to relative density, adds heavy 1% hydrochloric acid 20% alcoholic solution such as medical material weight, with the flow velocity of 1ml/min by 732 type strongly acidic styrene type cation exchange resins, 8BV is washed to neutrality, 80% ethanol 6BV remove impurity, 10BV1% ammonia 80% ethanol elution, eluant decompression and solvent recovery, drying, obtains uncaria total alkaloids; Wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%;
The preparation of Rhizoma Chuanxiong total alkaloids: get Ligusticum chuanxiong Hort, adds medical material weight 8 times amount 90% ethanol 70 DEG C of reflux, extract, 2 times, each 2 hours, and merging filtrate merges reclaim under reduced pressure, and dry, obtain Rhizoma Chuanxiong total alkaloids, wherein the content of ligustrazine is 0.23% ~ 0.24%;
The preparation of Radix Astragali total saponins: get Milkvetch Root, add 70% ethanol, 80 DEG C of reflux, extract, 1 hour of medical material weight 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1, with the flow velocity of 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 times of column volumes, again with 70% ethanol elution of 4 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
Get above-mentioned each effective site, add 10% starch, 90% alcohol granulation, to obtain final product.
Embodiment 4:
Prescription: the little peptide 3kg of Pheretima, uncaria total alkaloids 4kg, Rhizoma Chuanxiong total alkaloids 1.5kg, Radix Astragali total saponins 3kg
Method for making: get the little peptide of Pheretima, uncaria total alkaloids, Rhizoma Chuanxiong total alkaloids, Radix Astragali total saponins, mixing, add 10% starch, 90% alcohol granulation, to obtain final product.
Embodiment 5:
The routine Treating Patients of Angina Pectoris of the made Granules in Treating of Example 4 72.Therapeutic scheme: oral, one time 4,3 times on the one 4 weeks is a course for the treatment of.Clinical symptoms, sign, nitroglycerin reduction, ECG curative effect, blood lipid level etc. before and after treatment are observed after 2 courses for the treatment of.Result: cure 30 examples, take a turn for the better 23 examples, effective 13 examples, and invalid 6 examples, total effective rate is 91.7%.
Claims (4)
1. one kind is used for the treatment of the little peptide active component composition of Pheretima of cardiovascular disease, it is characterized in that by the little peptide of the Pheretima of extracting from Pheretima, from Ramulus Uncariae Cum Uncis, extract uncaria total alkaloids, extract from Rhizoma Chuanxiong in Rhizoma Chuanxiong total alkaloids and the Radix Astragali and extract that Radix Astragali total saponins forms, wherein, the weight proportion of each effective site is:
Pheretima little peptide 3 ~ 9 weight portion
Uncaria total alkaloids 6 ~ 8 weight portion
Rhizoma Chuanxiong total alkaloids 2 ~ 4 weight portion
Radix Astragali total saponins 6 ~ 8 weight portion.
2. the little peptide active component composition of Pheretima as claimed in claim 1, is characterized in that the weight proportion of each effective site is: Pheretima little peptide 6 weight portion, uncaria total alkaloids 8 weight portion, Rhizoma Chuanxiong total alkaloids 3 weight portion, Radix Astragali total saponins 6 weight portion.
3. the little peptide active component composition of Pheretima as claimed in claim 1 or 2, is characterized in that its preparation method comprises following steps:
(1) preparation of the little peptide of Pheretima: get Pheretima, suitable pulverizing, add 5 ~ 20 times of water gaging homogenate 5 ~ 15min, heated and boiled 5 ~ 30min, let cool to room temperature, hydrochloric acid is adopted to adjust pH to 2.0 ± 0.1, then the pepsin of amount of substrate 0.5% ~ 10% is added, enzymolysis 1 ~ 6h under 30 DEG C ~ 45 DEG C conditions, then sodium hydroxide solution is adopted to regulate pH to 7 ~ 10, add the trypsin of amount of substrate 0.5% ~ 10%, enzymolysis 1 ~ 6h under 45 DEG C ~ 55 DEG C conditions, stir constantly and regulate pH in 7 ~ 10 scopes, continue intensification after enzymolysis and boil 5 ~ 15min, let cool, centrifugal, collect supernatant, regulate pH6.5 ~ 7.0, add the active carbon of 1% ~ 3%, stir, filter, filtrate is that the hollow-fibre membrane of 3kD carries out ultrafiltration with retaining relative molecular mass, collect permeate, nanofiltration is carried out again to retain the ceramic membrane that relative molecular mass is 150D ~ 300D, collect non-permeate, concentrating under reduced pressure, dry, obtain the little peptide of Pheretima, wherein the content of the little peptide of Pheretima is 50% ~ 90%,
(2) preparation of uncaria total alkaloids: get Uncaria, add medical material weight 10 ~ 12 times of ethanol 75 ~ 80 DEG C of reflux, extract, 2 ~ 3 hours, filter, obtain filtrate, filtering residue repetitive operation twice, merge three parts of filtrates, decompression recycling ethanol to relative density is 1:1.8 ~ 1:2, add heavy 1% ethanol solution hydrochlorides such as medical material weight, with the flow velocity of 0.8 ~ 1ml/min by 732 type strongly acidic styrene type cation exchange resins, 6 ~ 8BV is washed to neutrality, ethanol 5 ~ 6BV remove impurity, 10 ~ 12BV1% ammonia ethanol elution, eluant decompression and solvent recovery, dry, obtain uncaria total alkaloids, wherein the content of Ramulus Uncariae cum Uncis alkali is 5% ~ 9%,
(3) preparation of Rhizoma Chuanxiong total alkaloids: get Ligusticum chuanxiong Hort, adds medical material weight 8 ~ 10 times amount ethanol 70 ~ 75 DEG C of reflux, extract, 2 times, each 1.5 ~ 2 hours, merging filtrate merges reclaim under reduced pressure, drying, obtains Rhizoma Chuanxiong total alkaloids, and wherein the content of ligustrazine is 0.23% ~ 0.24%;
(4) preparation of Radix Astragali total saponins: get Milkvetch Root, add ethanol 75 ~ 80 DEG C of reflux, extract, 1 ~ 1.5 hour of medical material weight 8 ~ 10 times amount, filter, obtain filtrate, filtering residue repetitive operation once, merge two parts of filtrates, decompression recycling ethanol to relative density is 1:1.8 ~ 1:2, add equivalent water-saturated n-butanol and extract three times, merge n-butanol fraction, reclaiming n-butyl alcohol to relative density is 1:1 ~ 1:1.2, with the flow velocity of 0.8 ~ 1ml/min by D101 macroporous resin, first with the distilled water eluting of 4 ~ 5 times of column volumes, again with the ethanol elution of 4 ~ 5 times of column volumes, collect eluent, reclaim under reduced pressure, dry Radix Astragali total saponins, wherein the content of astragaloside is 15% ~ 20%,
(5) get above-mentioned four parts of effective sites according to the weight proportion of each effective site, do not add adjuvant or add pharmaceutics customary adjuvant, make pharmaceutically acceptable dosage form.
4. the little peptide active component composition of the Pheretima described in claim 1 or 2 is used for the treatment of hypertension, high sticky blood, coronary heart diseases and angina pectoris.
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| CN1879669A (en) * | 2006-05-11 | 2006-12-20 | 上海交通大学 | Composition of ephedra total alkaloid and earthworm acid part and preparation method thereof |
| CN1969897A (en) * | 2005-11-23 | 2007-05-30 | 北京奇源益德药物研究所 | Pharmaceutical composition for treating cardiovascular and cerebrovascular diseases, preparation process and quality control method thereof |
| CN102266407A (en) * | 2011-08-05 | 2011-12-07 | 李运伦 | Traditional Chinese medicinal composition for treating hypertension and preparation method thereof |
| CN102920815A (en) * | 2012-11-26 | 2013-02-13 | 苏州法莫生物技术有限公司 | Application of active parts of gambir plant total alkaloids |
| CN104645077A (en) * | 2013-11-22 | 2015-05-27 | 万光瑞 | Traditional Chinese medicine preparation for preventing and treating ischemic cerebrovascular disease and preparation method of traditional Chinese medicine preparation |
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- 2016-01-13 CN CN201610020169.2A patent/CN105535924A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1969897A (en) * | 2005-11-23 | 2007-05-30 | 北京奇源益德药物研究所 | Pharmaceutical composition for treating cardiovascular and cerebrovascular diseases, preparation process and quality control method thereof |
| CN1879669A (en) * | 2006-05-11 | 2006-12-20 | 上海交通大学 | Composition of ephedra total alkaloid and earthworm acid part and preparation method thereof |
| CN102266407A (en) * | 2011-08-05 | 2011-12-07 | 李运伦 | Traditional Chinese medicinal composition for treating hypertension and preparation method thereof |
| CN102920815A (en) * | 2012-11-26 | 2013-02-13 | 苏州法莫生物技术有限公司 | Application of active parts of gambir plant total alkaloids |
| CN104645077A (en) * | 2013-11-22 | 2015-05-27 | 万光瑞 | Traditional Chinese medicine preparation for preventing and treating ischemic cerebrovascular disease and preparation method of traditional Chinese medicine preparation |
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