CN106039296A - Nattokinase composition and preparation method thereof - Google Patents
Nattokinase composition and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a nattokinase composition and a preparation method thereof. The composition is prepared from nattokinase, ligusticum wallichii and herba epimedii, and the fibrinolytic activity of the nattokinase in each gram of the composition is 200-400IU. The preparation method comprises the following steps: (1) preparing a nattokinase fermentation broth by utilizing bacillus subtilis which generates nattokinase, centrifuging, concentrating supernatant to obtain a nattokinase concentrate; (2) extracting herba epimedii with water, and concentrating the extracting liquid to prepare herba epimedii extract; (3) extracting ligusticum wallichii with water, and concentrating the extracting liquid to prepare ligusticum wallichii extract; and (4) mixing the ligusticum wallichii extract, the astragalus membranaceus extract and the nattokinase concentrate. The nattokinase composition is prepared by using nattokinase, ligusticum wallichii and herba epimedii as raw materials to achieve the synergistic effect, and has good effects of preventing thrombosis and thrombolysis.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition, be specifically related to natto kinase composition and preparation method thereof.
Background technology
Thrombotic disease as the high morbidity of middle-aged and elderly people, is one of maximum disease of death threats, current more than 60 years old
Old man reached more than 100,000,000, account for the 8.7% of population ratio, within 20 years from now on, China will enter aging society, thrombotic disease
Preventing and treating must cause enough attention.The whole world existing blood embolization patient about 15,000,000, the potential market of required Thrombolytic Drugs
It is worth about 2,000,000,000 dollars.The thrombolytic class medicine streptokinase that commonly uses on domestic market, urokinase, Plasminogen
Agent etc. all exist clearly disadvantageous: anti reperfusion, produce thromboembolism and hemorrhagic side effect again, and expensive.
Nattokinase (Nattokinase, NK) is a kind of Proteinkinase, is by receiving in soybean isoflavone by natto strain
A kind of serine protease that bean bacillus subtilis produces, has thrombus, reduces blood viscosity, improve blood circulation, soften and
Increase the effects such as blood vessel elasticity.Nattokinase is to be taught Xu Jian foreign firm doctor from 200 numerous food by Japan physiology in 1987
In first find and a kind of molecular weight of naming is far smaller than the protein of UK, SK, tPA, isoelectric point, IP is 8.7, and molecular weight is
27724, there is the strongest fibrinolytic capacity, and there is the activity of activatable elastic enzyme and uricase, phase after this
Pass research shows, nattokinase can effectively remove the thrombosis in blood of human body, is the protease that current thrombolytic vigor is the highest, and
And hypertension, aging, apoplexy, muscle spasm and cardiovascular and cerebrovascular disease are also had good curative effect, it is used as thrombolytic drug.And
And, nattokinase derives from the food natto of japanese traditional, and safety is good, simultaneously can be by intestinal absorption, effect lasts in vivo
Time is long, the most important be it can also recombinant type fibrinolysin activator in human activin, be allowed to gentle, improve blood constantly
The fibrinolytic of liquid, therefore, in recent years, medicine with nattokinase as main component, functional food, the grinding of food additive
Send out work progress the rapidest.For the application of nattokinase product, nutritionist's suggestion preventive dose daily is 1500
～2000IU, but this consumption also will bring higher cost so that the financial burden of thrombosis patients is heavier.
Summary of the invention
The technical problem to be solved in the present invention is to provide natto kinase composition and preparation method thereof.
The technical scheme that the present invention provides is natto kinase composition, and said composition includes nattokinase, Rhizoma Chuanxiong, Herba Epimedii
Composition；In every g compositions, the fibrinolytic of nattokinase is 200～400IU.
Above-mentioned nattokinase can be by the fermentation liquid of the bacillus subtilis producing nattokinase, it is also possible to is nattokinase lyophilizing
Powder.
Icariin is the main component of Herba Epimedii, and the content of icariin is 0.03～0.04mg/ in the composition
g。
Ligustrazine is the main component of Rhizoma Chuanxiong (formal name used at school: Ligusticum chuanxiong hort), in the composition
The content of ligustrazine is 0.02～0.03mg/g.
Present invention also offers the preparation method of above-mentioned natto kinase composition, comprise the following steps:
1) bacillus subtilis producing nattokinase is utilized to prepare nattokinase fermentation liquid, centrifugal, take supernatant concentration, received
Bean kinases concentrated solution；
2) by Herba Epimedii extracting in water, extracting solution is condensed into Herba Epimedii extractum；
3) by Rhizoma Chuanxiong extracting in water, extracting solution is condensed into Rhizoma Chuanxiong extractum；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed with nattokinase concentrated solution, to obtain final product.
The nattokinase of the present invention is from the tunning of the bacillus subtilis that can produce nattokinase, and bacillus subtilis is as one
Plant probiotic bacteria, market can directly be bought, such as Bacillus Subtilis AK (PN2003-210136, FERM P-
182391JP), Bacillus Subtilis MR141 (PN 08-154616, FERM P-14692JP), Bacillus
Subtilis IAM1026 (PN 5486467, FERM BP-6713US) etc., the decapacitation during the fermentation of these strains produces and receives
Outside bean kinases, metabolites at different levels all contain the composition useful to human body, therefore can serve as the strain of the present invention.The present invention
Preferably bacillus natto, it can produce the nattokinase of efficient fibrinolytic by solid or fluid matrix fermentation.
According to selected strain, its suitable condition of culture is selected to carry out fermentation culture, wherein it is possible to select solid fermentation
Method, makes bacteria suspension strain, is inoculated in solid medium, cultivates under appropriate conditions, will send out after fermentation ends
What ferment produced nattokinase dissolves to extract and obtains fermentation liquid, is centrifuged by fermentation liquid, supernatant concentration, obtains concentrated solution.
Owing to solid fermentation method is conventional method, the present invention does not elaborates herein.
Can also use solution fermentation, its concrete technology is as follows:
1) select nattokinase superior strain, on flat board, directly carry out separation screening, separate flat board preferred LB-agar
Sugar-fibrin plate: dissolve with 0.01M kaliumphosphate buffer (PB pH7.4) during the preparation of LB culture medium, when making flat board, will
Cryodesiccant Human Fibrinogen and thrombin are dissolved in wherein, the same to agarose-fibrin plate of its manufacture method.
2) according to the optimal culture condition of selected different strain, the nitrogen source in used fermentation medium, carbon source
Can select with inorganic salt, nitrogen source can select soy peptone, tryptone, yeast powder, caseinic acid hydrolysate with
And one or more in dry powdered soybean etc., the mass concentration of use is 0.5～about 4%；Carbon source can be glucose, wood
One or more in sugar, soluble starch, maltose, lactose and sucrose, its mass concentration is preferably left in 1.0～10.0%
Right；Inorganic salt can select KH2PO4About 0.2%, K2HPO4About 0.4%, MgSO4About 0.05%, CaCl2 about 0.02% etc..
According to currently preferred method, use bacillus natto to carry out fermentation preparation seed liquor, ferment to OD=0.3～
1.0, carry out liquid submerged fermentation, fermentation temperature 25～40 DEG C, fermentation period 12～35 with the inoculum concentration of 1.0～about 2.0%
Hour, pH is 6～8.
As preferably, the preparation method of natto kinase composition is:
1) utilize the bacillus subtilis producing nattokinase to prepare nattokinase fermentation liquid, be centrifuged with 3000～6000rpm, take
Clear liquid concentrates, and obtains nattokinase concentrated solution；
2) by Herba Epimedii extracting in water, extracting solution is condensed into the Herba Epimedii leaching that relative density at 60 DEG C is 1.10～1.20
Cream；
3) by Rhizoma Chuanxiong extracting in water, it is the Rhizoma Chuanxiong extractum of 1.20～1.45 that extracting solution is condensed into relative density at 60 DEG C；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed with nattokinase concentrated solution, to obtain final product.
Nattokinase fermentation liquid to be centrifuged, and rotating speed is excellent with 3000～6000rpm, can remove the big portion in fermentation liquid
Divide thalline and impurity.Owing to nattokinase is heat-sensitive substance, concentrate and general select room temperature to be concentrated by ultrafiltration, retaining point of ultrafilter membrane
Son amount is 50000～200000, and ultrafiltration pressure is 0.1～0.3MPa, rejection >=90%, and cycles of concentration regards natto in fermentation liquid
Depending on kinase whose fibrinolytic.
The preparation method of natto kinase composition it may also is that
1) utilize the bacillus subtilis producing nattokinase to prepare nattokinase fermentation liquid, be centrifuged with 3000～6000rpm, take
Clear liquid concentrates, and vacuum lyophilization obtains nattokinase lyophilized powder；
2) by Herba Epimedii extracting in water, it is the extractum of 1～1.20 that extracting solution is condensed into relative density at 60 DEG C, is dried,
Obtain Herba Epimedii dry powder；
3) by Rhizoma Chuanxiong extracting in water, it is the extractum of 1.20～1.45 that extracting solution is condensed into relative density at 60 DEG C, is dried,
Obtain Rhizoma Chuanxiong dry powder；
4) Herba Epimedii dry powder, Rhizoma Chuanxiong dry powder are mixed with nattokinase lyophilized powder, to obtain final product.
The compositions of the present invention can include that pharmaceutically acceptable adjuvant, preparation process routinely are made pharmaceutics and can be connect
Any regular dosage form being subject to, including capsule, tablet, granule, gel, slow releasing agent etc..Pharmaceutically acceptable adjuvant includes
Filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, preservative, substrate etc..Filler includes: form sediment
Powder, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.；Disintegrating agent includes: starch, pregelatinated form sediment
Powder, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linked carboxymethyl fiber
Element sodium etc.；Lubricant includes: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.；Suspending agent includes: polyethylene
Ketopyrrolidine, microcrystalline Cellulose, sucrose, agar, hydroxypropyl methyl cellulose etc.；Binding agent includes, starch slurry, polyvinyl pyrrole
Alkanone, hydroxypropyl methyl cellulose etc.；Sweeting agent includes: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.；Rectify
Taste agent includes: sweeting agent and various essence；Preservative includes: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters,
Benzalkonium bromide, acetic acid chloroethene are fixed, eucalyptus oil etc.；Substrate includes: PEG6000, PEG4000, insect wax etc..For enabling above-mentioned dosage form
Realize pharmacy of Chinese materia medica, need to add when preparing these dosage forms other adjuvant pharmaceutically acceptable (Fan Biting " pharmacy of Chinese materia medica ",
The adjuvant that in Shanghai Science Press December the 1st edition in 1997, each dosage form is recorded).
The day dose of natto kinase composition of the present invention is 1g, the wherein fibrinolytic of nattokinase in 1g compositions
Activity is only 200～400IU, well below conventional amount used, but fully achieves even more than fibrinolytic activity of nattokinase from natto
The thrombolytic effect of 2000IU, therefore can be greatly saved cost, alleviate the economic pressures of thrombotic disease patient.
All having certain thrombolytic effect according to prior art, Herba Epimedii and Rhizoma Chuanxiong, its active component is respectively excessive sheep
Icariin and ligustrazine, will reach preferable thrombolytic effect, and the consumption of ligustrazine is every day 0.1～0.5mg, the consumption of icariin
For every day 0.48～2mg.And take the compositions of the present invention, the day dose of ligustrazine is only 0.02～0.03mg, Herba Epimedii
The day dose of glycosides is only 0.03～0.04mg, well below conventional amount used.
The natto kinase composition of the present invention is made up of nattokinase, Rhizoma Chuanxiong and three kinds of components of Herba Epimedii, these three component
Play the effect of Synergistic, it is possible to extend the artery thrombosis time, reduce phlebothrombosis weight, the outer thromboembolism of acceleration bodies
Effect.
Detailed description of the invention
The present invention is further elaborated for specific examples below, but not as a limitation of the invention.
Nattokinase fermentation liquid described in following example prepares the most as follows:
1) select bacillus natto superior strain, on flat board, directly carry out separation screening, separate flat board preferred LB-agar
Sugar-fibrin plate: dissolve with 0.01M kaliumphosphate buffer (PB pH7.4) during the preparation of LB culture medium, when making flat board, will
Cryodesiccant Human Fibrinogen and thrombin are dissolved in wherein, the same to agarose-fibrin plate of its manufacture method.
2) in fermentation medium, nitrogen source selects soy peptone, and the mass concentration of use is 0.5～about 4%；Carbon source is
Glucose, its mass concentration is preferably 1.0～about 10.0%；Inorganic salt can select KH2PO4About 0.2%, K2HPO4About
0.4%, MgSO4About 0.05%, CaCl2 about 0.02%.Use bacillus natto to carry out fermentation preparation seed liquor, ferment to OD=
0.3～1.0, carry out liquid submerged fermentation, fermentation temperature 25～40 DEG C, fermentation period 12 with the inoculum concentration of 1.0～about 2.0%
～35 hours, pH is 6～8, obtains nattokinase fermentation liquid.
Embodiment 1
Said composition is made up of nattokinase, Rhizoma Chuanxiong and Herba Epimedii, and wherein, the consumption of nattokinase is with nattokinase fibrinolytic
Activity meter, in every g compositions, the fibrinolytic of nattokinase is 200IU；The consumption of Herba Epimedii in terms of icariin, said composition
The content of middle icariin is 0.03mg/g；The consumption of Rhizoma Chuanxiong is in terms of ligustrazine, and in said composition, the content of ligustrazine is
0.02mg/g。
The preparation method of said composition is:
1) nattokinase fermentation liquid is centrifuged with 3000rpm, by supernatant room temperature be concentrated by ultrafiltration, ultrafilter membrane retain molecule
Amount is 50000, and ultrafiltration pressure is 0.1MPa, obtains nattokinase concentrated solution；
2) by Herba Epimedii extracting in water, being filtered by extracting solution, filtrate is condensed into the excessive sheep that relative density at 60 DEG C is 1.10
Leaves of pulse plants extractum；
3) by Rhizoma Chuanxiong extracting in water, being filtered by extracting solution, filtrate is condensed into the Rhizoma Chuanxiong leaching that relative density at 60 DEG C is 1.20
Cream；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed in proportion with nattokinase concentrated solution, to guarantee every g compositions
The fibrinolytic of middle nattokinase is 200IU, and the content of icariin is 0.03mg, and the content of ligustrazine is 0.02mg.
Embodiment 2
Said composition is made up of nattokinase, Rhizoma Chuanxiong and Herba Epimedii, and wherein, the consumption of nattokinase is with nattokinase fibrinolytic
Activity meter, in every g compositions, the fibrinolytic of nattokinase is 400IU；The consumption of Herba Epimedii in terms of icariin, said composition
The content of middle icariin is 0.04mg/g；The consumption of Rhizoma Chuanxiong is in terms of ligustrazine, and in said composition, the content of ligustrazine is
0.03mg/g。
The preparation method of said composition is:
1) nattokinase fermentation liquid is centrifuged with 6000rpm, by supernatant room temperature be concentrated by ultrafiltration, ultrafilter membrane retain molecule
Amount is 200000, and ultrafiltration pressure is 0.3MPa, obtains nattokinase concentrated solution；
2) by Herba Epimedii extracting in water, it is the Herba Epimedii extractum of 1.20 that extracting solution is condensed into relative density at 60 DEG C；
3) by Rhizoma Chuanxiong extracting in water, it is the Rhizoma Chuanxiong extractum of 1.45 that extracting solution is condensed into relative density at 60 DEG C；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed with nattokinase concentrated solution, to guarantee natto in every g compositions
Kinase whose fibrinolytic is 400IU, the content of icariin is 0.04mg, the content of ligustrazine is 0.03mg.
Embodiment 3
Said composition is made up of nattokinase, Rhizoma Chuanxiong and Herba Epimedii, and wherein, the consumption of nattokinase is with nattokinase fibrinolytic
Activity meter, in every g compositions, the fibrinolytic of nattokinase is 300IU；The consumption of Herba Epimedii in terms of icariin, said composition
The content of middle icariin is 0.035mg/g；The consumption of Rhizoma Chuanxiong is in terms of ligustrazine, and in said composition, the content of ligustrazine is
0.025mg/g。
The preparation method of said composition is:
1) nattokinase fermentation liquid is centrifuged with 5000rpm, by supernatant room temperature be concentrated by ultrafiltration, ultrafilter membrane retain molecule
Amount is 100000, and ultrafiltration pressure is 0.2MPa, obtains nattokinase concentrated solution；
2) by Herba Epimedii extracting in water, it is the Herba Epimedii extractum of 1.15 that extracting solution is condensed into relative density at 60 DEG C；
3) by Rhizoma Chuanxiong extracting in water, it is the Rhizoma Chuanxiong extractum of 1.35 that extracting solution is condensed into relative density at 60 DEG C；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed in proportion with nattokinase concentrated solution, to guarantee every g compositions
The fibrinolytic of middle nattokinase is 300IU, the content of icariin is 0.035mg, the content of ligustrazine is 0.025mg.
Experimental example 1 is on the impact experiment of artery thrombosis in rat body
Take SD rat 70, body weight 180～220g, male and female half and half, be randomly divided into 7 groups, often group 10, male and female are each
5, respectively experimental group 1,2,3, matched group 1,2,3, and blank group.Experimental group 1,2,3 be administered respectively embodiment 1,
2, the compositions of 3, dosage is 1g/kg；Matched group 1 is administered nattokinase lyophilized powder (2000IU/g), matched group 2 is administered excessive
Sheep leaves of pulse plants extract (content of icariin is 0.04mg/g), matched group 3 are administered Rhizoma Chuanxiong extract, and (content of ligustrazine is
0.03mg/g), matched group dosage is 1g/kg；Blank group gives the normal saline of 1g/kg.Use gastric infusion,
Once a day, continuous seven days, last was administered latter 30 minutes, is drawn with 20% crow by rat but anaesthetizes, and peeled off rat carotid artery,
Distal end carries out arterial cannulation channel polygraph and adds measuring blood pressure, proximal part with 45% liquor ferri trichloridi soak
Filter paper bar parcel tremulous pulse wide for 0.5cm, the time when parcel to blood pressure reduces to 0 that records is the artery thrombosis time, record
Data, each group organize between t inspection statistics analysis.The results are shown in Table 2
Table 1 medicament composition capsule of the present invention is on the impact (X ± SD) of artery thrombosis in rat body
| Group | Number of cases (n) | Dosage (g/kg) | The artery thrombosis time (min) |
| Blank group | 10 | 1 | 10.55±1.54 |
| Experimental group 1 | 10 | 1 | 15.4±2.25ΔΔ |
| Experimental group 2 | 10 | 1 | 15.7±2.74Δ |
| Experimental group 3 | 10 | 1 | 15.25±1.30ΔΔ |
| Matched group 1 | 10 | 1 | 12.7±2.74Δ |
| Matched group 2 | 10 | 1 | 10.86±2.15 |
| Matched group 3 | 10 | 1 | 10.93±1.30Δ |
Note: compare with blank group, Δ p ＜ 0.05 Δ Δ p ＜ 0.01.
Experimental group 1～3 compares the artery thrombosis time that all can extend with matched group and blank group.Statistical result divides
Analysis shows, the natto kinase composition of the present invention has the effect extending the artery thrombosis time.
Experimental example 2 is on the impact experiment of venous thrombosis in rat body
Take SD rat 70, body weight 180～220g, male and female half and half, be randomly divided into 7 groups, often group 10, male and female are each
5, respectively experimental group 1,2,3, matched group 1,2,3, and blank group.Experimental group 1,2,3 be administered respectively embodiment 1,
2, the compositions of 3, dosage is 1g/kg；Matched group 1 is administered nattokinase lyophilized powder (2000IU/g), matched group 2 is administered excessive
Sheep leaves of pulse plants extract (content of icariin is 0.04mg/g), matched group 3 are administered Rhizoma Chuanxiong extract, and (content of ligustrazine is
0.03mg/g), matched group dosage is 1g/kg；Blank group gives the normal saline of 1g/kg.Use gastric infusion,
Once a day, continuous seven days, last was administered latter 30 minutes, is drawn with 20% crow by rat but anaesthetizes, and peeled off the total vein of rat cavity of resorption,
Ligature the total vein of cavity of resorption at left renal vein lower end line, open cavity of resorption total vein removal of thromboses after 3h and weigh, record data, each group
T inspection statistics analysis between organizing.The results are shown in Table 2
Table 2 medicament composition capsule of the present invention is on the impact (X ± SD) of venous thrombosis in rat body
| Group | Number of cases (n) | Dosage (g/kg) | Thrombosis weighs (g) |
| Blank group | 10 | 1 | 11.05±3.2 |
| Experimental group 1 | 10 | 1 | 5.08±2.71ΔΔ |
| Experimental group 2 | 10 | 1 | 5.11±1.89ΔΔ |
| Experimental group 3 | 10 | 1 | 4.94±2.91Δ |
| Matched group 1 | 10 | 1 | 7.11±1.89Δ |
| Matched group 2 | 10 | 1 | 10.59±2.18Δ |
| Matched group 3 | 10 | 1 | 10.43±3.07 |
Note: compare Δ p ＜ 0.05 Δ Δ p ＜ 0.01 with blank group.
As seen from the above table, the thrombus weight of the natto kinase composition of the present invention compares with blank group and has dropped
Low, and significantly lower than matched group 1～3.Analysis of statistical results shows, pharmaceutical composition of the present invention has reduction phlebothrombosis weight
Effect.
Experimental example 3 impact thromboclastic on rabbit inside and outside experiment
Taking healthy rabbits one, carotid artery is taken a blood sample, and static 2 hours, carefully divides with scalpel and takes 0.5g clot 70, weigh
After be respectively put in 7 test tubes, 10 clots put into by every test tube.Test tube is divided into 7 groups, respectively experimental group 1,2,3, comparison
Group 1,2,3, and blank group.Experimental group 1,2,3 is administered the compositions 0.14g+4.86g distillation of embodiment 1,2,3 respectively
Water；Matched group 1 adds nattokinase lyophilized powder (2000IU/g) 0.14g and 4.86g distilled water, matched group 2 adds Herba Epimedii
Extract (content of icariin is 0.04mg/g) 0.14g and 4.86g distilled water, matched group 3 add Rhizoma Chuanxiong extract (river
The content of rhizome of chuanxiong piperazine is 0.03mg/g) 0.14g and 4.86g distilled water, it is 0.14g that matched group is added to pharmaceutical quantities respectively；Blank
Matched group gives 5ml distilled water.Put into water-bath 3h in 37 DEG C of water baths, take out and weigh after sucking moisture, calculate by following equation
Dissolution rate: dissolution rate=(weight after weight-dissolving before dissolving)/dissolve front weight × 100%, the results are shown in Table 3.
The impact (X ± SD) that external thrombus is dissolved by table 3 medicament composition capsule of the present invention
| Group | Number of cases (n) | Dosage (g) | Dissolution rate (%) |
| Blank group | 10 | 5 | 11.8±2.89 |
| Experimental group 1 | 10 | 5 | 56.81±5.34Δ |
| Experimental group 2 | 10 | 5 | 57.43±2.45ΔΔ |
| Experimental group 3 | 10 | 5 | 58.6±4.81ΔΔ |
| Matched group 1 | 10 | 5 | 39.66±4.11Δ |
| Matched group 2 | 10 | 5 | 13.79±1.87Δ |
| Matched group 3 | 10 | 5 | 13.15±2.39 |
Note: compare Δ p ＜ 0.05 Δ Δ p ＜ 0.01 with blank group,
The natto kinase composition of the present invention compares with blank group all can increase external thrombus dissolution rate, and thrombolytic effect
Fruit is substantially better than matched group.Analysis of statistical results shows, the natto kinase composition of the present invention has the outer thromboembolism of acceleration bodies
Effect.
Claims (7)
1. natto kinase composition, it is characterised in that: said composition includes nattokinase, Rhizoma Chuanxiong and Herba Epimedii composition；Every g combination
In thing, the fibrinolytic of nattokinase is 200～400IU.
Natto kinase composition the most according to claim 1, it is characterised in that: in said composition, the content of icariin is
0.03～0.04mg/g.
Natto kinase composition the most according to claim 1, it is characterised in that: in said composition, the content of ligustrazine is
0.02～0.03mg/g.
4. the preparation method of the natto kinase composition according to any one of claims 1 to 3, it is characterised in that: include following
Step:
1) bacillus subtilis producing nattokinase is utilized to prepare nattokinase fermentation liquid, centrifugal, take supernatant concentration, obtain natto and swash
Enzyme concentrated solution；
2) by Herba Epimedii extracting in water, extracting solution is condensed into Herba Epimedii extractum；
3) by Rhizoma Chuanxiong extracting in water, extracting solution is condensed into Rhizoma Chuanxiong extractum；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed with nattokinase concentrated solution, to obtain final product.
The preparation method of natto kinase composition the most according to claim 4, it is characterised in that: comprise the following steps:
1) utilize the bacillus subtilis producing nattokinase to prepare nattokinase fermentation liquid, be centrifuged with 3000～6000rpm, take supernatant
Concentrate, obtain nattokinase concentrated solution；
2) by Herba Epimedii extracting in water, it is the Herba Epimedii extractum of 1.10～1.20 that extracting solution is condensed into relative density at 60 DEG C；
3) by Rhizoma Chuanxiong extracting in water, it is the Rhizoma Chuanxiong extractum of 1.20～1.45 that extracting solution is condensed into relative density at 60 DEG C；
4) Herba Epimedii extractum, Rhizoma Chuanxiong extractum are mixed with nattokinase concentrated solution, to obtain final product.
The preparation method of natto kinase composition the most according to claim 5, it is characterised in that: comprise the following steps:
1) utilize the bacillus subtilis producing nattokinase to prepare nattokinase fermentation liquid, be centrifuged with 3000～6000rpm, take supernatant
Concentrate, vacuum lyophilization, obtain nattokinase lyophilized powder；
2) by Herba Epimedii extracting in water, it is the extractum of 1.10～1.20 that extracting solution is condensed into relative density at 60 DEG C, is dried,
To Herba Epimedii dry powder；
3) by Rhizoma Chuanxiong extracting in water, it is the extractum of 1.20～1.45 that extracting solution is condensed into relative density at 60 DEG C, is dried, obtains
Rhizoma Chuanxiong dry powder；
4) Herba Epimedii dry powder, Rhizoma Chuanxiong dry powder are mixed with nattokinase lyophilized powder, to obtain final product.
The preparation method of natto kinase composition the most according to claim 4, it is characterised in that: described bacillus subtilis is for receiving
Bean bacillus.
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| CN107692200A (en) * | 2017-09-08 | 2018-02-16 | 江苏大学 | A kind of Nattokinase epiphysin composition for improving sleep and preparation method thereof |
| CN109260461A (en) * | 2017-07-17 | 2019-01-25 | 张乐泷 | A kind of Chinese medicine composition for treating cardiovascular disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109260461A (en) * | 2017-07-17 | 2019-01-25 | 张乐泷 | A kind of Chinese medicine composition for treating cardiovascular disease |
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