Summary of the invention
Technical problem to be solved by this invention is existing Cornu Cervi side (in high spirit preparation) active ingredient extracting method is improved, a kind of active ingredient content height is provided, the preparation solid preparation is convenient, and the quality of the pharmaceutical preparations is stable, the method for preparing Cornu Cervi square preparation (in high spirit preparation) of determined curative effect.
The preparation method of the Chinese medicine preparation of treatment chronic heart failure disclosed by the invention (the Cornu Cervi square preparation has another name called in high spirit preparation) comprises the following steps:
1, raw material weight percentage ratio prescription
Colla cornus cervi 1%-5% or Cornu Cervi tablet 8%-25%, Herba Epimedii 15%-40%, Fructus Ligustri Lucidi 15%-40%, Fructus Psoraleae 10%-30%, Fructus Corni 8%-25% and Pericarpium Citri Reticulatae 8%-25%;
2, preparation method
Get Colla cornus cervi or Cornu Cervi tablet and pulverize, cross 80 mesh sieves, standby; Getting all the other components in the prescription adds the water that 10-30 doubly measures and decocts 2-3 time for 90 ± 10 ℃, filter, collecting decoction, being concentrated into relative density is 1.04-1.05 (room temperature), the 14000-16000rps high speed centrifugation, getting supernatant concentration to relative density is 1.10-1.14 (90 ℃), spray drying or add a small amount of dextrin spray drying and get extract powder; Above-mentioned harts horn carbon is mixed with extract powder, and adding pharmaceutic adjuvant formulation method is routinely made solid orally ingestible;
Or get in the prescription except that Colla cornus cervi each raw material and add the water that 10-30 doubly measures and decoct 2-3 time filtration, collecting decoction for 90 ± 10 ℃, the 14000-16000rps high speed centrifugation, get supernatant concentration, adding preservative agent, correctives or the Colla cornus cervi after molten are made mixture according to a conventional method.
Each oral formulations of the present invention comprises granule and mixture etc., takes icariin content every day greater than 54mg/g, and psoralen is greater than 42mg/g.
The inventor finds that through repetition test the active ingredient that prior art adopts the water extract-alcohol precipitation method to extract in the Cornu Cervi side can make main active ingredient icariin and psoralen lose more.With Cornu Cervi side's (in high spirit preparation) each component (except the Colla cornus cervi) decocting liquid 90 ℃ the time is example with 70% precipitate with ethanol and high speed centrifugation method of the present invention remove impurity respectively, and the active ingredient result is compared as follows table:
Impurity-removing method icariin content reduces psoralen content and reduces the minimizing of isopsoralen content
(%)?????????????(%)???????????????(%)
70% precipitate with ethanol 14.72 20.79 20.84
High speed centrifugation 9.41 14.17 16.91
The above results shows that decoction and alcohol sedimentation technique makes active ingredient, and particularly the icariin content loss is more, and adopts decocting liquid of the present invention through centrifugal impurity removal process, can effectively reduce the loss of active ingredient, extracts active ingredient to greatest extent.
In addition compared with prior art, the extract of the inventive method acquisition is convenient to the preparation of solid preparation.When extract of the present invention was prepared into solid preparation, usually extracting solution must the spray-dried extract powder that becomes, and granulate, thereby supplementary product consumption in the minimizing preparation reduces clinical taking dose again.If the sample liquid behind the water extract-alcohol precipitation is carried out spray drying, very easily sticking wall is difficult to obtain dried powder, and loss is very big, even obtain powder, because of hygroscopicity is very strong, also is difficult to make granule, makes troubles for the preparation of solid preparation.And the present invention through ultracentrifugal medicinal liquid because to remove impurity comparatively thorough, concentrate, after the spray drying, can obtain dry loose powdered, the back yield rate height of granulating can be suitable for large-scale production.
Chinese medicine preparation of the present invention is specially adapted to treat Chinese medical discrimination and belongs to the chronic heart failure of suffering from a deficiency of the kidney.
Carried out toxicity test with Chinese medicine preparation of the present invention: acute toxicity, the maximum tolerated dose of irritating the stomach mice is crude drug 212g crude drug/kg; Long term toxicity, the high, medium and low dosage group of granule compares with blank group, and the overt toxicity reaction is not seen in administration in continuous six months.Prove that preparation of the present invention is safe in utilization.
By Chinese medical discrimination suffer from a deficiency of the kidney standard and the diagnostic criteria of doctor trained in Western medicine Framingham heart failure, carry out clinical research with the mixture and the granule of extract preparation of the present invention to going into to organize patient, every group 40 example, the result shows:
Preparation of the present invention can significantly improve CHF patient's NYHA cardiac functional grading (obvious effective rate 47%, total effective rate 87%) and the scoring of Lee heart failure, and its curative effect and captopril and digoxin matched group are similar substantially.Simultaneously we can also significantly improve CHF patient's symptom integral of suffering from a deficiency of the kidney, thereby the prompting Chinese medicine especially has one's own knack aspect CHF patient's clinical symptoms improving on treatment CHF really.On the index that ultrasoundcardiogram detected, can see, compare with the digoxin matched group, after taking preparation of the present invention, can significantly improve CHF patient's IVST, PWT, EF, CO, FS, LVMI (P<0.05), especially the reduction of IVST, PWT, LVMI, in conjunction with the improvement of patient's cardiothoracic ratio, illustrate that we can strengthen myocardium shrinkage function (improving the biological function of cardiac muscle), can suppress or reverse the development of cardiac remodeling again simultaneously.To secular reparation characteristic index, improvement is in various degree arranged all from the hemodynamic index of short-term.This also meets the direction of present heart failure treatment very much: not only improve symptom, improve the quality of living, the more important thing is the mechanism at cardiac remodeling, prevent or delay the development of cardiac remodeling, thereby reduce the mortality rate and the admission rate of heart failure.
Comprehensively above-mentioned, as to adopt the inventive method to prepare Cornu Cervi square preparation, active ingredient content height, preparation is convenient, determined curative effect, this method is particularly suitable for industrialization production.
The specific embodiment
Embodiment 1 in high spirit particulate preparation
1. get Colla cornus cervi 2.5kg and pulverize, cross 80 mesh sieves, standby.
2. get Herba Epimedii 25kg, Fructus Ligustri Lucidi 20kg, Fructus Psoraleae 25kg, Fructus Corni 15kg, Pericarpium Citri Reticulatae 10kg, the water that at every turn adds 10 times of amounts decocts three times for 90 ℃, first, each 1 hour for the second time, filter the 3rd half an hour, merges three times decocting liquid, being concentrated into relative density is 1.04-1.05 (room temperature), the 16000rps high speed centrifugation, getting supernatant concentration to relative density is 1.10-1.14 (90 ℃), adds an amount of dextrin, spray drying gets extract powder, add the Cornu Cervi rubber powder again, an amount of dextrin and correctives, mixing is with 85% alcohol granulation that contains 2%PVP, dry, get granule 36kg, granulate, packing.Wherein icariin content is 2.22mg/g, and psoralen is 0.76mg/g.
Embodiment 2 in high spirit particulate preparations
1. get Colla cornus cervi 3kg and pulverize, cross 80 mesh sieves, standby.
2. Herba Epimedii 30kg, Fructus Ligustri Lucidi 30kg, Fructus Psoraleae 20kg, Fructus Corni 15kg, Pericarpium Citri Reticulatae 15kg add 100 ℃ of extractions of water, totally three times, first, second time each 1 hour, filters the 3rd half an hour, merge three times decocting liquid, the 14000rps high speed centrifugation, getting supernatant concentration to relative density is 1.10-1.14 (90 ℃), spray drying gets extract powder, add Cornu Cervi rubber powder and an amount of dextrin, mixing, the dried grain that extrudes, get granule 30kg, packing.Wherein icariin content is 3.58mg/g, psoralen 0.82mg/g.
Embodiment 3 in high spirit particulate preparations
1. get Colla cornus cervi 2.2kg and pulverize, cross 100 mesh sieves, standby.
2. get Herba Epimedii 25kg, Fructus Ligustri Lucidi 22kg, Fructus Psoraleae 18kg, Fructus Corni 13kg, Pericarpium Citri Reticulatae 13kg, add water and decoct secondary, each 1 hour for 100 ℃, filter, merge the secondary decocting liquid, be concentrated into an amount of, the 16000rps high speed centrifugation, getting supernatant concentration to relative density is 1.12 (90 ℃), adds an amount of dextrin, mixing, spray drying gets extract powder, adds Cornu Cervi rubber powder and an amount of dextrin, fluidized granulation, get granule 30kg, packing.Wherein icariin content is 2.82mg/g, and psoralen is 0.70mg/g.
Embodiment 4 icariin HPLC assays
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-water (55: 45) is a mobile phase; Detect wavelength 270nm.
It is an amount of that the preparation precision of reference substance solution takes by weighing the icariin reference substance, adds methanol and make the solution that every 1ml contains 0.0204mg.
It is an amount of that the sample that embodiment 1 makes is got in the preparation of need testing solution, is ground into fine powder, and precision takes by weighing 0.5g, put in the tool plug conical flask, the accurate methanol 25ml that adds claims to decide weight, supersound extraction 60min, taking-up room temperature to be chilled to claims to decide weight, supplies the weight that subtracts mistake with methanol, shake up, filter, get filtrate, promptly.
Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
The result: icariin content is 2.22mg/g.
Embodiment 6 psoralen assays
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-water (45: 55) is a mobile phase; Detect wavelength 246nm.
It is an amount of that the preparation precision of reference substance solution takes by weighing the psoralen reference substance, adds methanol and make the solution that every 1ml contains psoralen 0.014mg.
It is an amount of that the sample that embodiment 1 makes is got in the preparation of need testing solution, is ground into fine powder, and precision takes by weighing 0.5g, puts in the tool plug conical flask, the accurate methanol 50ml that adds puts into supersonic generator and extracts 60min, takes out room temperature to be chilled to, and filters, the accurate filtrate 20ml that draws, evaporate to dryness, residue add water 20ml makes dissolving, uses chloroform extraction 5 times, each 20ml, combined chloroform liquid, reclaim under reduced pressure is to doing, residue makes dissolving with methanol, moves in the 25ml measuring bottle gradation washing, add methanol to scale, shake up, promptly.
Accurate respectively reference substance solution and each the 5 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
Psoralen content is 0.76mg/g as a result.
Embodiment 7 stability tests
It is indoor that embodiment 1 is made sample (aluminium plastic membrane packing) placement, and temperature 20-35 ℃, relative humidity 40%-80% deposited three months.Investigate sample character, discriminating, moisture, granularity, melting, assay and microbial limit every month one time, the results are shown in Table.The result shows that this product is through trimestral preliminarily stabilised investigation, and every index is all up to specification, tentatively shows this product steady quality.
Standing time project |
0 month (August 6) |
January (JIUYUE 9 days) |
February (October 7) |
March (November 8) |
Character |
Brown particle, bitter in the mouth |
Brown particle, bitter in the mouth |
Brown particle, bitter in the mouth |
Brown particle, bitter in the mouth |
Differentiate |
(1) Colla cornus cervi feature speckle (2) should be detected and Fructus Psoraleae feature speckle should be detected |
????+ ????+ |
????+ ????+ |
????+ ????+ |
????+ ????+ |
Check |
Moisture granularity melting |
4.31 3.2 is qualified |
4.40 3.1 is qualified |
4.35 3.5 is qualified |
4.51 3.0 is qualified |
Assay |
Excessive sheep flower bud glycosides content psoralen content |
????2.22 ? ????0.76 |
????2.18 ? ????0.81 |
????2.24 ? ????0.74 |
????2.27 ? ????0.78 |
Limit test of microbe |
Antibacterial mycete escherichia coli |
103/g<10 do not detect |
56/g<10 do not detect |
100/g<10 do not detect |
179/g<10 do not detect |
The preparation of embodiment 8 in high spirit mixture
Get Cornu Cervi tablet 0.75kg, Herba Epimedii 1.5kg, Fructus Ligustri Lucidi 1.2kg, Fructus Psoraleae 0.9kg, Fructus Corni 0.75kg, Pericarpium Citri Reticulatae 0.75kg, the water that at every turn adds 10 times of amounts decocts three times for 100 ℃, first, second time each 1 hour, the 3rd half an hour, filter collecting decoction, 14000rps high speed centrifugation, get supernatant concentration to an amount of, add an amount of sucrose, sodium benzoate, ethyl hydroxybenzoate, boil, volume is adjusted to 5300ml, packing, sterilization promptly.Wherein icariin content is 1.25mg/ml, and psoralen is 0.26mg/ml.