CN101502558A - Method for preparing astragalus total saponin injection from cyclodextrin composition - Google Patents
Method for preparing astragalus total saponin injection from cyclodextrin composition Download PDFInfo
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Abstract
The invention relates to a preparation method of a membranous milkvetch root preparation, in particular to the method for preparing an injection of total saponins of membranous milkvetch root by using a cyclodextrin composition. The problem of unstable quality control in the existing manufacturing process of membranous milkvetch root injection is solved. The method comprises the steps of taking the total saponins of membranous milkvetch root to dissolve in ethanol, taking the cyclodextrin composition to dissolve in 7000-9000ml water for injection, adding the ethanol solution of the total saponins of membranous milkvetch root into the cyclodextrin solution, heating and stirring at 60 DEG C till no alcohol smell; adding 12-hydroxystearic acid polyethylene glycol ester, stirring till clarification of the solution; adding sodium chloride, stirring, dissolving, processing with activated carbon, filtering, removing the carbon; fine filtering, filling, sealing and sterilizing to obtain the sodium chloride injection of the total saponins of membranous milkvetch root.
Description
Technical field
The present invention relates to a kind of preparation method of formulation of astragalus root, be specially a kind of method of preparing astragalus total saponin injection from cyclodextrin composition.
Background technology
The Radix Astragali is the conventional Chinese medicine material, have to support unit, strengthening body resistance and dispelling evil, nourish heart promote blood circulation, the effect of invigorating spleen to remove dampness, the modern clinical hepatitis that is widely used in the motive is deficient, blood vessels become silted up resistance viral myocarditis, cardiac insufficiency and stagnation of dampness due to deficiency of the spleen etc.Radix Astragali injection is a kind of common dosage form, and Radix Astragali injection is at the clinical treatment that is used for obstruction of qi in the chest and cardialgia disease in a large number, determined curative effect.After deliberation, the main effective ingredient of its cardiovascular system activity is the Radix Astragali saponin class, pharmacological evaluation proof it and Radix Astragali injection therapeutic equivalence.That the Radix Astragali injection of present clinical use remains is not high at content, lack the reliable quality standard, wayward, the clinical use of drug effect can cause reactions such as patient allergy, the not high deficiency of safety in utilization.
The preparation method kind of existing Radix Astragali injection is many, and for example the patent No. is 031419275 to disclose a kind of astragaloside injection preparation and preparation method thereof.In this patent application, provide that a kind of content is clear and definite, quality controllable, the preparation method of the Radix Astragali total saponins injection of stable performance.Raw material has used Radix Astragali total saponins, and cosolvent (especially be combined as the best with propylene glycol, ethanol, and consumption is 0.5：1-1：0。5), the content of Radix Astragali total saponins is 5-50 milligram/bottle, and in this patent specification, the medicine that is logged into this application manufacturing is the buchu Radix Astragali injection.And this injection has carried out experimentation to the protective effect of dog acute myocardial infarction.Be that feedstock production becomes injection directly, can improve the content of Radix Astragali total saponins, increase quality controllability, make more effective, the safety of formulation of astragalus root with Radix Astragali total saponins through extracting purification.But Radix Astragali total glycosides is insoluble in water, and need seek suitable method when it is prepared into injection increases its dissolubility.The patent No. is the preparation method that the patent of 200510095308.x has been announced a kind of astragaloside cyclodextrin inclusion, and main technical schemes is that astragaloside is made cyclodextrin clathrate, is equipped with adjuvant again according to the needs of dosage form and is prepared into all kinds of different dosage forms.But Radix Astragali total saponins complicated component, remove astragaloside and also contain other saponins, there is qualitative difference between various saponin again, experiment shows, only prepare cyclodextrin clathrate earlier with said method and prepare in the process of Radix Astragali total saponins injection again, cyclodextrin is through super-dry and be dissolved in after depositing a period of time again and will occur in the process of water that enclose is incomplete, significant loss big, be difficult for making settled solution, solution instability, be prone to problem such as precipitation.
Summary of the invention
The present invention provides a kind of method of preparing astragalus total saponin injection from cyclodextrin composition in order to solve the quality control problem of unstable that exists in the existing Radix Astragali injection manufacture process.
The present invention realizes by following technical scheme, and a kind of method of preparing astragalus total saponin injection from cyclodextrin composition, step comprise,
(1) get the 2.4-3.0g Radix Astragali total saponins and be dissolved in the 500-1000ml ethanol, get cyclodextrin composite and be dissolved in the 7000-9000 milliliter water for injection, the Radix Astragali total saponins alcoholic solution is added in the cyclodextrin solution, 60 ℃ of heated and stirred are to there not being the alcohol flavor;
(2) add polyethyleneglycol-12-hydroxy stearin 50-200g, be stirred to the solution clarification;
(3) add sodium chloride 90g, stirring and dissolving, charcoal treatment, filtering decarbonization;
(4) replenish water for injection to 10000 milliliter, fine straining, embedding, sterilization.
Promptly get the astragalus total-saponin sodium chloride injector that Radix Astragali total saponins content is the 0.24-0.3 mg/ml.With 0.27 mg/ml is the best.
In the above-mentioned preparation method, the used cyclodextrin composite of enclose is that beta-schardinger dextrin-and HP-are according to the blended compositions of mass ratio 1:1-2.The used mass ratio of Radix Astragali total saponins and cyclodextrin composite is 1:4-10 during enclose.The working concentration of polyethyleneglycol-12-hydroxy stearin is 0.5%-2% or 5mg/ml-20mg/ml.
With different rate of charges, adopt paddling process to carry out enclose, beta-schardinger dextrin-and HP-are according to mass ratio 1:1,1:2,1:3,1:4,1:5,1:10 makes up and carries out enclose with Radix Astragali total saponins 1:1, the sampling and measuring Radix Astragali total saponins, Astragaloside content, the result with beta-schardinger dextrin-and HP-according to mass ratio 1:1, the compositions rate of charge of 1:2 is more suitable, 1:3,1:4,1:5, the compositions of 1:10 feeds intake and 1:1, the compositions rate of charge of 1:2, content differs and is not clearly, the compositions that 1:1-2 is described feeds intake and can reach best enclose effect, changes of contents is not obvious along with the increase of ingredient proportion, the declaratives cyclodextrin is excessive for feeding intake, have neither part nor lot in enclose, invalid composition, so we have determined that beta-schardinger dextrin-and HP-are the best of breed rate of charge according to mass ratio 1:1-2.Radix Astragali total saponins, Astragaloside content and inclusion rate are as shown in the table in the different compositions rate of charge clathrates.0.2735mg/ml, be 0.0921mg/ml.
Radix Astragali total saponins and Astragaloside content and inclusion rate in the different compositions rate of charge clathrate of table 1
The cyclodextrin composite ratio determine the back we again with beta-schardinger dextrin-and HP-compositions according to mass ratio 1:2, press 1:1 with Radix Astragali total saponins, 2:1,4:1,8:1,10:1,20:1 carries out enclose, survey Radix Astragali total saponins and Astragaloside content in the clathrate, the result is more suitable with the rate of charge that Radix Astragali total saponins and cyclodextrin composite are pressed 1:4-10,1:1-2 feeds intake, it is lower to record content, illustrate that enclose is incomplete, 1:20 feeds intake and 1:4-10 compares, content differs and is not clearly, illustrating feeds intake between the 1:4-10 can reach best enclose effect, changes of contents is not obvious along with the increase of ingredient proportion, and the declaratives cyclodextrin is excessive for feeding intake, and has neither part nor lot in enclose, invalid composition, so we have determined that the optimum charging ratio of Radix Astragali total saponins and cyclodextrin composite is 1:4-10.Radix Astragali total saponins and the contained quantity of astragaloside and inclusion rate are as shown in the table in the different rate of charge clathrates.
Radix Astragali total saponins and contained quantity of astragaloside and inclusion rate in the different rate of charge clathrates of table 2
We make astragalus total-saponin sodium chloride injector with the Radix Astragali total saponins clathrate that said method makes according to the preparation method of injection, carry out reserved sample observing, after storing 6 months, there is 1 bottle slight muddiness to occur in 100 bottles, illustrate that the astragalus total-saponin sodium chloride injector that adopts cyclodextrin clathrate to make still is undesirable, we have added the polyethyleneglycol-12-hydroxy stearin solubilizing agent again in above-mentioned technology for this reason, and its consumption and stability are tested.
We select beta-schardinger dextrin-and the HP-compositions according to mass ratio 1:2 for use, Radix Astragali total saponins and this cyclodextrin composite are that 1:4 carries out enclose by mass ratio, and the concentration of pressing 0.1%-5% adds polyethyleneglycol-12-hydroxy stearin, be prepared into astragalus total-saponin sodium chloride injector, find in the preparation process that the polyethyleneglycol-12-hydroxy stearin of adding 0.5%-2% concentration has obviously improved the content of astragaloside, and under identical storage requirement, place and observed in 6 months, the injection of all adding polyethyleneglycol-12-hydroxy stearin does not all become turbid as a result, is transparent supernatant liquid.The content that adds variable concentrations polyethyleneglycol-12-hydroxy stearin Radix Astragali total saponins and astragaloside is as shown in the table.
Radix Astragali total saponins and Astragaloside content in the table 3 variable concentrations polyethyleneglycol-12-hydroxy stearin
For the further stability of the prepared injection of checking the present invention, we select same batch of injection to test under 60 ℃, 18-26 ℃, 0-4 ℃, 4576lx illumination condition.Result such as following table:
Table 5 injection stability test result
| Condition | Time (my god) | Clarity | Differentiate | The content of Radix Astragali total saponins (mg/ml) | The content of astragaloside (mg/ml) |
| 60℃ | 0 30 90 180 | Clarification clarification clarification clarification | Positive | 0.2742 0.2739 0.2737 0.2737 | 0.0931 0.0921 0.0918 0.0917 |
| 18-26℃ | 0 30 90 180 | Clarification clarification clarification clarification | Positive | 0.2744 0.2740 0.2738 0.2737 | 0.0933 0.0927 0.0919 0.0915 |
| 0-4℃ | 0 30 90 180 | Clarification clarification clarification clarification | Positive | 0.2738 0.2739 0.2735 0.2731 | 0.0929 0.0925 0.0916 0.0918 |
| 4576lx | 0 30 90 180 | Clarification clarification clarification clarification | Positive | 0.2743 0.2739 0.2731 0.2727 | 0.0934 0.0927 0.0916 0.0909 |
Can find that through a large amount of tests Radix Astragali total saponins is insoluble in water, the dissolubility of effective ingredient and the stability of solution significantly improved after Radix Astragali total saponins was made clathrate and added the solubilizer polyethylene glycol 12-hydroxy stearic acid ester with the compositions of two kinds of cyclodextrin.
With produce the Radix Astragali total saponins cyclodextrin inclusion earlier described in the background technology, the technology of utilizing clathrate to prepare injection is again compared, the astragalus total-saponin sodium chloride injector that directly makes with the inventive method, good stability, the medicament contg height, good effect, toxic and side effects is low.The clear liquid of the astragalus total-saponin sodium chloride injector that the present invention is made was placed 6 months under 4-8 ℃, room temperature and 60 ℃ of temperature conditions, and any variation does not take place the solution character, and Radix Astragali total glycosides content is surveyed in sampling, does not change, and medicinal liquid is stable, reliable.
The influence of the rat heart muscle ischemia that we also cause pituitrin astragalus total-saponin sodium chloride injector of the present invention is studied.
Be subjected to the reagent thing: astragalus total-saponin sodium chloride injector, the present invention makes; Radix Astragali injection, Shanghai Hefeng Pharmaceutical Co., Ltd. produces, and outward appearance is brown, and every ml contains crude drug 2g; The pituitrin injection: Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd produces, and faces with preceding that to be made into 2U/ml with normal saline standby.More than two kinds of medicines all stablize 30 minutes posterior veins and instil in the every index that is write down.
Animal: healthy male SD rat, the The 2nd Army Medical College Experimental Animal Center provides, 50, body weight is a 350-450 gram, be divided at random 5 groups as follows:
Grouping number of animals (only) dosed administration approach
The 10 5mi/Kg intravenous drips of normal saline group
The 10 2g/Kg intravenous drips of Radix Astragali injection group
The 10 1.5mg/Kg intravenous drips of Radix Astragali total saponins injection low dose group
Dosage group 10 5mg/Kg intravenous drips in the Radix Astragali total saponins injection
The 10 15mg/Kg intravenous drips of Radix Astragali total saponins injection high dose group
Experimental technique:
Rat is with 3% pentobarbital sodium 45mg/Kg intraperitoneal injection of anesthesia.Separate left femoral vein and keep somewhere polyethylene tube in order to administration.It is subcutaneous to insert extremity with needle electrode, and record arm's length standard limbs II leads electrocardiogram (ECG-6511 type electrocardiograph, Shanghai Photoelectricity Medical electron Instrument Co., Ltd.).If any electrocardiographic abnormality then this animal give it up.Stablize 30min, intravenous drip is subjected to the reagent thing.In femoral vein 10 seconds, inject pituitrin 5U/Kg behind the 30min, after the injection immediately each time point II of continuous record lead electrocardiogram (chart drive speed 50mm/s, voltage 10mm * 1).Draw 15sec, 30sec behind the medicine, 60sec, 2min, 5min, 10min, 20min and 30min time point then and carry out statistical disposition, observe the ST section and raise degree change.Statistical analysis: each time period ST section is raised degree and is deducted basic value (△ value) in the experimental result, represents with x ± S, does the time T check relatively, and P＜0.05 is judged as the significant difference with significance.
Experimental result:
(1) general electrocardiogram changes behind the intravenous injection pituitrin
The result shows: general electrocardiogram mainly shows as the ST section and raises rapidly behind the rat intravenous injection pituitrin, and T ripple step-down is flat or be inverted, and prompting has myocardial ischemia.Arrhythmia shows the chamber of taking place frequently early, and also visible height is to atrioventricular block fully.
(2) influence that damaging ST section is raised by the reagent thing
The result shows: bringing out the ST section that causes of damage with injection of pituitrin, to raise degree be judge index, basic, normal, high three dosage groups of Radix Astragali total saponins injection and positive control drug Radix Astragali injection all can significantly reduce anesthesia SD rat intravenous injection pituitrin and cause that the ECG ST section that myocardial damage causes raises, wherein obvious with the middle and high dosage group of Radix Astragali total saponins effect, show dose-effect relationship preferably.
Conclusion:
Astragalus total-saponin sodium chloride injector 1.5mg/Kg, 5mg/Kg and 15mg/Kg intravenous injection all can reduce the ST section that SD rat myocardium from injury that pituitrin brings out causes and raise degree; Though small dose group △ ST section and normal saline group comparison absolute value have reduction, but the most of the time, Duan Wei reached statistically-significant difference, and that middle and high dosage group and normal saline group comparison △ ST section are raised degree difference is remarkable, illustrate that Radix Astragali total saponins has better effect to preventing myocardium blood damage, and certain dose-effect relationship is arranged.
The specific embodiment
Embodiment 1
1. take by weighing Radix Astragali total saponins 2.4g (wherein containing astragaloside 0.8g), be dissolved in about 500ml ethanol; (beta-schardinger dextrin-and HP-are according to mass ratio 1: 1-2) be dissolved in 7000 milliliters of waters for injection, the Radix Astragali total saponins alcoholic solution is added in the cyclodextrin solution, 60 ℃ of heated and stirred are to there not being the alcohol flavor to get the 24g cyclodextrin composite;
2. add polyethyleneglycol-12-hydroxy stearin 50g, be stirred to the solution clarification;
3. add sodium chloride 90g, stirring and dissolving adds 0.1% charcoal treatment, filtering decarbonization;
4. supply water for injection to 10000ml, fine straining, embedding, the sterilization of pressurizeing of 100ml/ bottle, 30min, check, packing, promptly.
The Radix Astragali total saponins injection that present embodiment makes, sampling and measuring, Radix Astragali total saponins content are 0.2397g/ml, Astragaloside content is 0.0787mg/ml.Be placed on 4-8 ℃, room temperature and 60 ℃ of temperature conditions were placed 12 months down, take out the back and observe, medicinal liquid clear does not still have any precipitate as a result, and significant change does not take place the medicinal liquid character.Significant change does not take place in the content that sampling records Radix Astragali total glycosides and astragaloside.
Embodiment 2
1. take by weighing Radix Astragali total saponins 2.7g (wherein containing astragaloside 0.9g), be dissolved in about 650ml ethanol; Get 12g cyclodextrin composite (beta-schardinger dextrin-and HP-are according to mass ratio 1:1-2) and be dissolved in 8000 milliliters of waters for injection, the Radix Astragali total saponins alcoholic solution is added in the cyclodextrin solution, 60 ℃ of heated and stirred are to there not being the alcohol flavor;
2. add polyethyleneglycol-12-hydroxy stearin 100g, be stirred to the solution clarification;
3. add sodium chloride 90g, stirring and dissolving adds 0.1% charcoal treatment, filtering decarbonization;
4. supply water for injection to 10000ml, fine straining, embedding, the sterilization of pressurizeing of 100ml/ bottle, 30min, check, packing, promptly.
The Radix Astragali total saponins injection that present embodiment makes, sampling and measuring, Radix Astragali total saponins content are 0.2735mg/ml, Astragaloside content is 0.0921mg/ml.Be placed on 4-8 ℃, room temperature and 60 ℃ of temperature conditions were placed 12 months down, take out the back and observe, medicinal liquid clear does not still have any precipitate as a result, and significant change does not take place the medicinal liquid character.Significant change does not take place in the content that sampling records Radix Astragali total glycosides and astragaloside.
Embodiment 3
1. take by weighing Radix Astragali total saponins 3g (wherein containing astragaloside 1g), be dissolved in about 750ml ethanol; Get 24g cyclodextrin composite (beta-schardinger dextrin-and HP-are according to mass ratio 1:1-2) and be dissolved in 9000 milliliters of waters for injection, the Radix Astragali total saponins alcoholic solution is added in the cyclodextrin solution, 60 ℃ of heated and stirred are to there not being the alcohol flavor;
2. add polyethyleneglycol-12-hydroxy stearin 200g, be stirred to the solution clarification;
3. add sodium chloride 90g, stirring and dissolving adds 0.1% charcoal treatment, filtering decarbonization;
4. supply water for injection to 10000ml, fine straining, embedding, the sterilization of pressurizeing of 100ml/ bottle, 30min, check, packing, promptly.
The Radix Astragali total saponins injection that present embodiment makes, sampling and measuring, Radix Astragali total saponins content are 0.3112g/ml, Astragaloside content is 0.1022mg/ml.Be placed on 4-8 ℃, room temperature and 60 ℃ of temperature conditions were placed 12 months down, take out the back and observe, medicinal liquid clear does not still have any precipitate as a result, and significant change does not take place the medicinal liquid character.Significant change does not take place in the content that sampling records Radix Astragali total glycosides and astragaloside.
Claims (4)
1, a kind of method of preparing astragalus total saponin injection from cyclodextrin composition is characterized in that: step comprises,
(1) get the 2.4-3.0g Radix Astragali total saponins and be dissolved in the 500-1000ml ethanol, get cyclodextrin composite and be dissolved in the 7000-9000 milliliter water for injection, the Radix Astragali total saponins alcoholic solution is added in the cyclodextrin solution, 60 ℃ of heated and stirred are to there not being the alcohol flavor;
(2) add polyethyleneglycol-12-hydroxy stearin 50-200g, be stirred to the solution clarification;
(3) add sodium chloride 90g, stirring and dissolving, charcoal treatment, filtering decarbonization;
(4) replenish water for injection to 10000 milliliter, fine straining, embedding, sterilization,
Promptly get the astragalus total-saponin sodium chloride injector that Radix Astragali total saponins content is the 0.24-0.3 mg/ml.
2, the method for preparing astragalus total saponin injection from cyclodextrin composition according to claim 1 is characterized in that: cyclodextrin composite is that beta-schardinger dextrin-and HP-are according to the blended compositions of mass ratio 1:1-2.
3, the method for preparing astragalus total saponin injection from cyclodextrin composition according to claim 1 is characterized in that: the used mass ratio of Radix Astragali total saponins and cyclodextrin composite is 1:4-10 during enclose.
4, the method for preparing astragalus total saponin injection from cyclodextrin composition according to claim 1 is characterized in that: Radix Astragali total saponins content is 0.27 mg/ml.
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| CN104116777A (en) * | 2013-04-26 | 2014-10-29 | 成都力思特制药股份有限公司 | Polyoxyethylene-660-12-hydroxy stearate containing astragalus membranaceus injection drug preparation and preparation method thereof |
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| CN100367969C (en) * | 2003-07-29 | 2008-02-13 | 上海博泰医药科技有限公司 | Injection formulation of astragalus root saponin and its preparation mehod |
| CN1785202B (en) * | 2004-12-06 | 2010-12-01 | 天津天士力制药股份有限公司 | Medicinal composition for treating cardiovascular disease and its preparation method |
| CN100393319C (en) * | 2005-11-08 | 2008-06-11 | 南京工业大学 | Astragaloside cyclodextrin clathrate, its prepn. and preparing mehtod |
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| CN104116777A (en) * | 2013-04-26 | 2014-10-29 | 成都力思特制药股份有限公司 | Polyoxyethylene-660-12-hydroxy stearate containing astragalus membranaceus injection drug preparation and preparation method thereof |
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