CN1695714A - Combination of medication for treating osteoporosis and preparation method - Google Patents
Combination of medication for treating osteoporosis and preparation method Download PDFInfo
- Publication number
- CN1695714A CN1695714A CN 200510020867 CN200510020867A CN1695714A CN 1695714 A CN1695714 A CN 1695714A CN 200510020867 CN200510020867 CN 200510020867 CN 200510020867 A CN200510020867 A CN 200510020867A CN 1695714 A CN1695714 A CN 1695714A
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- Prior art keywords
- fructus psoraleae
- total flavones
- radix puerariae
- pharmaceutical composition
- coumarins
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Abstract
A Chinese medicine for treating osteoporosis is prepared from psoralea fruit and pueraria root. Its preparing process is also disclosed.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition for the treatment of osteoporosis, particularly, it is to be the pharmaceutical composition that feedstock production forms by Fructus Psoraleae, Radix Puerariae.
Background technology
Osteoporosis is one of modal senile disease, and nearly in the world at present 200,000,000 people suffer from osteoporosis, and its sickness rate has leapt to the 7th of the various commonly encountered diseases in the world, and the medical expert has classified it as 21 century influences one of serious disease of human health.Osteoporotic pathogenic process is slow, progressive, more hidden, be difficult to be aware, direct result is a fracture, can bring out because of wound, when serious even in walking, cough and ridge awl or Hip Fracture take place when carrying weight, the vertebra folding can cause persistence neuralgia or paralysis, several cones are subsided and can be caused the splanchnocoel volume to reduce, compressing internal organs and influence its function.
The osteoporotic medicine of treatment mainly is divided into following three classes at present, anti-bone absorption class, and main activity by the inhibition osteoclast reduces CaE, as estrogens, bis phosphoric acid salt, calcitonin; Short sclerotin forms class: as fluoride, anabolic hormone etc.; Promote bone mineralising class: as vitamin D, calcium and boron preparation etc.
It is existing historical for many years that Hormone Replacement Therapy is used to prevent and treat women's osteosporosis after menopause, increases the danger that uterus carcinoma takes place and use estrogen to have for a long time separately.Selective estrogen receptor modulators (Selgtive Estrogen ReceptorModulators, SEMRs), be a class have aromatic ring structure can with the bonded chemical compound of estrogen receptor, wherein raloxifene (Raloxifene) has gone on the market and has tried out in osteoporosis, though do not increase carcinogenic danger, but along with the expansion of clinical research scale and the prolongation of time, the discovery raloxifene is prevented and treated osteoporotic effect and not obvious.The bis phosphoric acid salt can be by reducing the effect of osteoclast, induces osteoclast to transfer to die and reduce bone resorption, alleviates osteodynia, improves osseous tissue sign, and being mainly used in treatment has the enhanced disease of obvious bone resorption.Take this type of medicine for a long time and can suppress the bone mineralising, behind the first generation phosphate general therapeutic 1-2, the bone amount no longer includes progress after being increased to a stationary value, occurs untoward reaction such as hyperphosphatemia and osteomalacia after the small number of patients medication.In second and third,, phosphoric acid salt had untoward reaction such as gastrointestinal upset, low blood calcium and hypophosphatemia and erythra in generation.The calcium preparation kind reaches more than 200 kinds, calcium source difference, and the physicochemical property difference, the absorbance difference of various preparations is big, and uncertain therapeutic efficacy is cut, and l threonic acid and calcium amino acid chelate curative effect are better, but cost an arm and a leg.Vitamin D and derivant thereof are accumulated in vivo easily, and poisoning by vitamin D easily takes place, and show as hypercalcemia, higher urinary calcium even internal organs calculus, and taking this type of medicine should have strict indication, and results of regular determination urine calcium.
Treat the most uncertain therapeutic efficacy of osteoporotic Chinese medicine at present clinically and cut, it is unreliable to act on, and dosage form is backward relatively, mostly is one, secondary dosage form, and day clothes dosage is big, and takes often, brings great inconvenience to the patient.Application number 02154159.0 discloses the medicine of a kind of prevention, treatment osteoporosis, and it is the medicament that is formed for feedstock production by natural plants (Chinese medicine) such as Rhizoma Drynariae, Herba Cistanches, Fructus Psoraleae, Herba Epimedii, Radix Panacis Quinquefoliis.Since the material choice difference, the consumption difference, method of treatment rule of treatment difference, the Fructus Psoraleae of still having no way of at present, Radix Puerariae are the report of the osteoporotic medicine of treatment of feedstock production.
Summary of the invention
Technical scheme of the present invention has provided a kind of pharmaceutical composition for the treatment of osteoporosis, and it is to be that feedstock production forms medicament by Fructus Psoraleae, Radix Puerariae.Another technical scheme of the present invention has provided this preparation of drug combination method and purposes.
The pharmaceutical composition of treatment osteoporosis provided by the invention is the medicament that is prepared from by the following weight proportion raw material:
6～9 parts of Fructus Psoraleaes, 9～15 parts of Radix Puerariaes.
Preferred scheme is the medicament that it is prepared from by the following weight proportion raw material:
10 parts of Fructus Psoraleaes, 10 parts of Radix Puerariaes.
Be by Fructus Psoraleae total coumarins, Fructus Psoraleae total flavones, Radix Puerariae total flavones is an active component, adds the medicament that acceptable accessories or complementary composition are prepared from.
Wherein, described Fructus Psoraleae total coumarins is in psoralen, and percentage by weight is not less than 24%, Fructus Psoraleae total flavones is in Coryfolin, and percentage by weight is not less than 26%; Radix Puerariae total flavones is in puerarin, and percentage by weight is not less than 70%.
The pharmaceutical composition of described treatment osteoporosis is the medicament that is prepared from by the following weight proportion raw material: 1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones.
Separate because of Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones are difficult, therefore, it is extracted simultaneously.
Further, it is to be prepared from medicament by the following weight proportion raw material: 1 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1 part of Radix Puerariae total flavones.
Described medicament is: tablet, capsule, pill, granule, slow releasing preparation.
Described slow releasing preparation is prepared from by following weight proportion raw material and adjuvant:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones, slow releasing preparation adjuvant: 1～10 part.
Described slow releasing preparation adjuvant is: a kind of or two or more mixing arbitrarily in ethyl cellulose EC, hydroxypropyl methylcellulose HPMC, Sulisi surelease, ethylmethylcellulose, hydroxypropyl cellulose, cellulose acetate, glue connection polyvidone, the polyacrylic resin.
Preferred version, slow releasing preparation is prepared from by following weight proportion raw material and adjuvant:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones, 0.5～10 part of EC, 0.2～1.0 part of HPMC.
Further, it is to be prepared from by following weight proportion raw material and adjuvant:
1 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1 part of Radix Puerariae total flavones, 0.5 part of EC, HPMC0.5 part.
The method of the pharmaceutical composition of preparation treatment osteoporosis provided by the invention comprises the steps:
A, get Fructus Psoraleae, add organic solvent extraction, Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture;
B, get Radix Puerariae, with 30～90% alcohol reflux, ethanol extract decompression recycling ethanol, filter, by adsorption resin column, with 30%～90% ethanol elution, collect ethanol elution, reclaim ethanol, drying promptly gets Radix Puerariae total flavones;
C, with the Fructus Psoraleae total coumarins of b, c step preparation, Radix Puerariae total flavones by following weight proportion, add acceptable accessories or complementary composition and be prepared into dosage form pharmaceutically commonly used:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones.
Wherein, the described reflux, extract, concentration of alcohol of b step is 30% ethanol; The concentration of ethanol elution is 70% ethanol.
Fructus Psoraleae is recorded in Lei Gong's Treatise on Preparation and Broiling of Materia Medica the earliest, cures mainly five kinds of strain and seven kinds of impairment, and the bone marrow wound loses.All on the books in " property of medicine opinion ", " Kaibao Bencao ", " Bencao Tujing ", the Compendium of Material Medica.Li Shizhen (1518-1593 A.D.) is said; Fructus Psoraleae is sayed its merit also; Broken solid paper belongs to fire, can receive the sword gods, and the fire of pericardium is communicated with fire from the gater of life, so Yuanyang is firm, bone marrow enriches, but Fructus Psoraleae ball strengthening bone and muscle, beneficial vigour." experience rear " record, Fructus Psoraleae are fried and are the end, and the heat wine clothes are controlled lumbago due to renal deficiency down, and are wonderful." prescription of peaceful benevolent dispensary " record, blue or green pretty young woman's ball is controlled a little less than the deficiency of kidney-QI, lumbago as the folding, under the light vinegar soup of married woman, informal dress strengthening bone and muscle, the arteries and veins of invigorating blood circulation.The Radix Puerariae beginning is stated from " herbal classic ", classifies middle product as, and sweet in the mouth is flat, " main all numbness ".The numbness that Radix Puerariae is controlled is the numbness of " apathy ", and its clinical manifestation is similar to the symptom due to the osteoporosis." Japan hanako materia medica " has the clearly record of Radix Puerariae " removing blood stasis ", and Radix Puerariae can promoting blood circulation to remove obstruction in the collateral." Handbook of Prescriptions for Emergencies " record, kidney pain in the lumbar region pain is given birth to the pharynx juice that Radix Puerariae is chewed, and getting effect is to end." Zhu Liang spring medication experience " record, " with invigorating function of kidney and strengthening bone Zhi Qiben " must use Radix Puerariae, thinks that Radix Puerariae can " treatment rheumatism involving the bone, spasmolytic is promoted blood circulation ".Record GEGEN TANG in " the classical prescription fax " of Hu Xishu, he thinks " lumbar muscle strain, the bright of this nothings exterior syndrome levied, the effect of controlling with we repeatly ".Always once with the GEGEN TANG plus-minus treatment atrophic debility of bones, obtain clinical efficacy preferably recklessly.Modern study shows that Radix Puerariae total flavones can effectively be prevented and treated osteoporosis.Research such as Tan Zhilong Radix Puerariae is to the influence of removal ovary rat biochemical markers of bone metabolism, and the result shows that Radix Puerariae can increase estradiol (E2), suppresses bone resorption, promote bone formation, thus the osteoporotic effect of performance treatment.
By the pharmacodynamics test explanation, medicine material Fructus Psoraleae of the present invention, Radix Puerariae compatibility, can effectively treat osteoporosis, reach the effect of Synergistic, wherein further active component Fructus Psoraleae total coumarins, Fructus Psoraleae total flavones and the Radix Puerariae active component Radix Puerariae total flavones proportioning of Fructus Psoraleae are used, the effect of same performance Synergistic, and because effective site is determined, reduce taking dose, be convenient to preparation and be prepared into the slow releasing drugs of tcm type, make the slow release effect fully be suitable for indication of the present invention, reach long-acting purpose; Fructus Psoraleae and Radix Puerariae are the kind of integration of edible and medicinal herbs in addition, and said preparation not only has therapeutical effect, also have functions of prevention and health care, reach the control combination jointly, the purpose for the treatment of both the principal and the secondary aspects of a disease at the same time.To sum up, medicine material Fructus Psoraleae total coumarins of the present invention, Fructus Psoraleae total flavones, Radix Puerariae total flavones compatibility use, and reach the effect of Synergistic, and drug effect is clear and definite, steady quality, and controllability is strong, provides a kind of new selection for clinical.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill and the customary means of this area.
The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example, all technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
The specific embodiment
The preparation of embodiment 1 medicine capsule of the present invention
Get raw material Fructus Psoraleae 9kg, Radix Puerariae 9kg, add 50% alcohol reflux respectively, reclaim ethanol, be condensed into thick extractum, the adjuvant that adds 15-50% is granulated, and is encapsulated.
The preparation of embodiment 2 medicinal tablets of the present invention
Fructus Psoraleae 6kg, Radix Puerariae 9kg add 90% alcohol reflux respectively, reclaim ethanol, are condensed into thick extractum, and the adjuvant that adds 15-50% is granulated tabletting.
The preparation of embodiment 3 medicinal granules of the present invention
Fructus Psoraleae 9kg, Radix Puerariae 15kg add 30% ethanol percolation respectively, reclaim ethanol, are condensed into thick extractum, and the adjuvant that adds 15-50% is made granule.
The preparation of embodiment 4 medicament pellets of the present invention
Fructus Psoraleae 6kg, Radix Puerariae 15kg add 50% alcohol reflux respectively, reclaim ethanol, spray drying, and the adjuvant that spray powder adds 10-50% is round as a ball in the high-efficiency coating pot, screening 20-40 purpose micropill.
The extracting method of embodiment 5 medicine materials of the present invention
Fructus Psoraleae adds 6 times of amount 70% alcohol heating reflux and extracts 3 times, each 1h, merging filtrate is when being concentrated into concentration and being 0.5g/ml, add 3 times of water gaging precipitations, left standstill 24 hours, and abandoned supernatant, get precipitation, add the n-hexane extraction defat of 3 times of amounts, discard normal hexane, precipitation vacuum decompression drying gets Fructus Psoraleae total flavones and the blended Fructus Psoraleae of Fructus Psoraleae total coumarins and treats osteoporotic effective site.
Radix Puerariae adds 9 times of amount 30% alcohol reflux 3 times, each 1 hour, merging filtrate, being concentrated into concentration is 0.5g/ml, amount with 60 times of resin bed volumes is added on SP207 macroporous resin capital, medicinal liquid PH is 5-6, the rate of adsorption is 2BV/h, with 70% ethanol elution of 4 times of resin bed volumes, collects eluent, being concentrated into relative density is 1.02-1.05, carry out spray drying, charging rate is 50-50ml/min, and inlet temperature is 135-145 ℃, leaving air temp is 75-85 ℃, obtains the osteoporotic effective site Radix Puerariae total flavones of kudzuvine root for treating.
The preparation of embodiment 6 medicine capsules of the present invention
Get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 1kg, Radix Puerariae total flavones 1kg, the adjuvant that adds 10-50% is granulated, 60 ℃ of dryings, and granulate, encapsulated, promptly.
The preparation of embodiment 7 medicinal tablets of the present invention
Get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 5kg, Radix Puerariae total flavones 1kg, the adjuvant that adds 10-50% is granulated tabletting.
The preparation of embodiment 8 medicinal granules of the present invention
Get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 5kg, Radix Puerariae total flavones 3kg, the adjuvant that adds 10-50% is made granule, packing.
The preparation of embodiment 9 medicament pellets of the present invention
Get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 4kg, Radix Puerariae total flavones 3kg, the adjuvant that adds 10-50% is granulated, and the adjuvant that adds 10-50% is round as a ball in the high-efficiency coating pot, screening 20-40 purpose micropill.
Embodiment 10 provides the preparation method of slow releasing preparation of the present invention
Slow-release micro-pill: get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 2kg, Radix Puerariae total flavones 1kg, add a grain powder of 15% and roll into 20-40 purpose micropill, the Sulisi with 15% carries out coating, makes the micropill 7-15% that increases weight, and promptly gets slow-release micro-pill.
Slow releasing tablet: get Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture 1kg, Radix Puerariae total flavones 1kg, granulate, tabletting, the Sulisi with 15% carries out coating, makes tablet weightening finish 5-10%, promptly gets slow releasing tablet.
The selection test of embodiment 11 slow releasing pharmaceutical supplementary product kinds of the present invention
Ethyl cellulose (EC) is insoluble framework material, and chemical property is stable, and is nontoxic, parmacodynamics-less activity, and the insoluble framework material for commonly used also can be used as sustained release coating liquid usefulness, has been widely used in the preparation of slow releasing agent had good sustained release effect; Hydroxypropyl methylcellulose (HPMC) is a hydrophilic gel matrix material, meeting water or Digestive system expands, form the gel barrier and have the character of control medicine stripping, the hydroxypropyl methylcellulose consumption after a little while, share with ethyl cellulose and to carry out coating, the effect of certain porogen is arranged, thereby improve the result of extraction of medicine.Sulisi (surelease) is that a kind of use ethyl cellulose is as controlled-release material, the aqueous dispersion that contains film former, plasticizer and stabilizing agent, be a kind of full water coating system that is simple and easy to usefulness, and the release of medicine is not subjected to the influence of pH value, can be used for the coating of granule, piller and tablet.Sulisi is avoided the screening difficulty and the time of autogamy coating prescription, reduces operation easier, enhances productivity.
Solid dispersion method
Press the prescription proportioning of table 1, the skeleton agent is dissolved in respectively in 80% the ethanol, add intermediate by recipe quantity and make it dissolving, reclaim ethanol, 50 ℃ of dryings 1 hour, porphyrize, encapsulated, measure the release in vitro degree of Radix Puerariae total flavones.Release in vitro degree assay method is for changeing the basket method, rotating speed 100 ± 1r/min, 37 ± 0.5 ℃ of temperature, release medium is the phosphate buffer of 900mlpH=7.4, under these conditions, in different time points sampling 5ml (adding the 5ml dissolution medium after the sampling immediately), add dilute hydrochloric acid and transfer pH to 5-6, water bath method adds the ultrasonic 10min of 5ml chloroform, discard chloroform solution, residue adds the 10ml dissolve with methanol again, filters, and filtrate adds methanol constant volume to 25ml, measure absorbance in the 250nm place, calculate the cumulative release degree of Radix Puerariae total flavones.
Table 1 solid dispersion method prescription table
| The prescription number | Intermediate (%) | ????EC(％) | HPMC K50(％) |
| ????1 ????2 ????3 | ????95 ????95 ????95 | ????5 ????-- ????2.5 | ????-- ????5 ????2.5 |
The cumulative release degree (%) of Radix Puerariae total flavones (n=3) in the slow releasing capsule of table 2 different auxiliary material
As can be seen from Table 2, when supplementary product consumption is 5%, promptly reach about 80%, fail to reach the effect of slow release, consider the adjuvant ratio is increased, in the hope of obtaining slow release effect stripping in the 1st hour.
Table 3 supplementary product consumption examination prescription table
| The prescription number | Intermediate (%) | ????EC(％) | ????HPMC K50(％) |
| ????1 ????2 ????3 ????4 ????5 ????6 | ????80 ????80 ????80 ????50 ????50 ????50 | ????20 ????-- ????10 ????50 ????-- ????25 | ????-- ????20 ????10 ????-- ????50 ????25 |
The cumulative release degree (%) of Radix Puerariae total flavones (n=3) in the slow releasing capsule of table 4 different auxiliary material
From table 3,4 as can be seen, have only when adjuvant increases to 50%, can obtain slow release effect, day clothes dosage is 3.1g, can not satisfy the needs of formulation dosage, and supplementary product consumption is excessive, and increases cost, so consider to select for use other preparation method.
The matrix type slow-release micro-pill
Press the prescription of table 5, fine drug powder is mixed with blocker, cross 100 mesh sieves, get partially mixed powder and place the coating pan molding, the residue mixed powder strengthens molding, makes the skeleton micropill, and is encapsulated, measures the cumulative release degree of Radix Puerariae total flavones by above-mentioned condition.
Table 5 matrix type micropill prescription table
| The prescription number | Pharmaceutical composition (%) | ????EC(％) | ????HPMC(％) |
| ????1 ????2 ????3 ????4 ????5 ????6 | ????50 ????50 ????50 ????30 ????30 ????30 | ????50 ????-- ????25 ????70 ????-- ????35 | ????-- ????50 ????25 ????-- ????70 ????35 |
Table 6 matrix type micropill Radix Puerariae total flavones cumulative release degree measurement result
From table 5,6 as can be seen, the burst size of medicine reduces with the increase of blocker consumption, and when the consumption of adjuvant during up to 2.3 times of medicated powder, also do not reach ideal slow release effect, increase the consumption of adjuvant again, slow release effect preferably be might reach, but taking dose and production cost also increased simultaneously.And film controlling type micropill supplementary product consumption is less relatively, so transformation of the way film controlling type micropill.
The film controlling type slow-release micro-pill
The film controlling type micropill is after medicine is at first made the ball heart, in the extended release coatings of ball core outsourcing different-thickness, to reach the purpose that drug slow is released effect again, but also has the advantages that to improve outward appearance, taste, increase medicine stability.
The preparation of plain ball
This pharmaceutical composition is met water and is easily bondd, and the pill operating difficulties can add appropriate amount of auxiliary materials and be improved, and selects starch cheap and easy to get for use, and with the pharmaceutical composition mixing, with the general pill heart of partially mixed powder, residue bisque layer strengthens round as a ball, and sieve is got 20-40 purpose micropill.
With not commensurability starch and medicated powder mixing, cross 100 mesh sieves, place coating pan, make binding agent with 80% ethanol, get the general ball heart of partially mixed powder, the residue powder strengthens, and is evaluation criterion with the complexity of operation, the roundness of ball, determines the consumption of adjuvant with this.
When supplementary product consumption was 15%, processing ease carried out, and micropill has only a small amount of adhesion than rounding between ball, so the consumption of starch is decided to be 15%.
Matrix sustained release tablet
Pharmaceutical composition is mixed by table 7 with framework material, cross 100 mesh sieves, use 70% alcohol granulation, cross 14 mesh sieves, 50 ℃ of dryings 1 hour, with 16 mesh sieve granulate, adding fluidizer and lubricant mixing are pressed into the slice, thin piece of 0.4g weight, measure release as stated above.
The different proportioning tables of table 7 pharmaceutical composition with adjuvant
| The prescription number | Pharmaceutical composition (%) | ????EC(％) | HPMC K50(％) |
| ????1 ????2 ????3 ????4 ????5 ????6 | ????80 ????80 ????80 ????50 ????50 ????50 | ????20 ????-- ????10 ????50 ????-- ????25 | ????-- ????20 ????10 ????-- ????50 ????25 |
Table 8 matrix sustained release tablet drug release determination result
As can be seen from Table 8, increase along with skeleton agent consumption, rate of release slows down, but when skeleton agent consumption is 20%, fail to reach the purpose of slow release, and when supplementary product consumption is 50%, though can reach certain slow release effect, but supplementary product consumption is very big, and day clothes dosage surpasses 3g, can not satisfy the requirement of preparation.So make the film controlling type slow releasing tablet into, in the hope of reducing the consumption of adjuvant.
The film controlling type slow releasing tablet
The selection of filler
Extract powder was mixed 100 mesh sieves with the starch of different proportion, place coating pan, spray into 50% ethanol and rotate granulation, continuation turns to drying in pot, tabletting, the outward appearance of more prepared various slice, thin pieces, hardness, compressibility, the result indicates, every performance indications no significant difference of prepared tablet is considered from the angle that reduces cost, simplifies technology, can not add filler.
The examination of lubricant kind
Lubricant can increase particulate flowability, reduces tablet weight variation, has the characteristics of anti-adhesive, avoids sticking, makes tablet surface bright and clean attractive in appearance, can also reduce the frictional force between tablet and nib, is easy to slice, reduces the wearing and tearing of punch die.
It is an amount of to get dried granule, add 0.5% magnesium stearate, 0.5% Pulvis Talci and 0.5% micropowder silica gel mixing respectively, measure particulate angle of repose by the fixed funnel method, the result indicates, after adding magnesium stearate, this granule angle of repose is 37.1 °, and fluidizer is better than other two kinds of lubricants, so select magnesium stearate as lubricant.
The examination of lubricant quantity
It is an amount of to get dried granule, adds the magnesium stearate mixing of grain amount 0.25%, 0.5%, 1% respectively, measures angle of repose; When lubricant quantity be particle weight 0.5% the time, existing good mobility.
Various preparation methoies comprehensively compare
From preparation technology's operation easier, the consumption of adjuvant and release conditions comprehensively compare above-mentioned various preparation methoies, select the dosage form that is fit to this prescription.Comparative result sees Table 9.
The various preparation method comprehensive comparison of table 9
| Dosage form | Preparation method | Supplementary product consumption | The operation difficulty or ease | Outward appearance | Release |
| The capsule tablet | Solid dispersion method microskeleton type ball film controlling type matrix type film controlling type | ??＞50％ ??＞70％ ??＜25％ ??＞50％ ??＜10％ | Easily difficult easy than difficulty or ease | Graininess is round, smooth surface any surface finish surface-brightening | Slow quickly, it is slowly fast to meet the requirements, and meets the requirements |
As can be seen from Table 9, this prescription can be made film controlling type slow-release micro-pill and slow releasing tablet, but the preparation tablet is easier than micropill operation, supplementary product consumption still less, dose is little, cost is low, meets the requirement of big production, so be made into film controlling type slow releasing tablet optimum selection.
The method of quality control of embodiment 12 pharmaceutical preparatioies of the present invention
One, the content of Fructus Psoraleae Coumarins and bavachin in the employing determined by ultraviolet spectrophotometry preparation:
The preparation of need testing solution
Get 10 of drug sustained-release tablets of the present invention, porphyrize is got 0.25g, and accurate the title decides, add 20ml chloroform supersound extraction 30min, filter, filtrate volatilizes, and residue adds dissolve with methanol, be settled to 100ml, the accurate 1ml that draws adds methanol and is diluted to 50ml, as the need testing solution of measuring total coumarins content.Get 0.25g, the accurate title, decide, and adds 20ml methanol supersound extraction 30min, filters, and adds methanol constant volume to 100ml, and the accurate 1ml that draws adds methanol and is diluted to 50ml, as the need testing solution of measuring general flavone content.
The selection of absorbing wavelength
The Coumarins composition has common furan mother nucleus structure in the Fructus Psoraleae, and uv absorption is consistent substantially, is reference substance so select psoralen for use, and the content of total coumarins is in psoralen.It is an amount of to get the psoralen reference substance, adds dissolve with methanol, carries out UV scanning at the 200-400nm place, and the mensuration maximum absorption wavelength is 243nm, also is 243nm with the maximum absorption wavelength of need testing solution UV scanning, is 243nm so select the mensuration wavelength of total coumarins.
The flavones ingredient uv absorption has bigger difference in the Fructus Psoraleae, can't be with the absorbing wavelength of certain single flavone as measuring wavelength, test by prerun, after the NaOH colour developing, the equal red shift of the absworption peak of flavone is to 440nm, and need testing solution UV scanning maximum absorption wavelength is also at 440nm, so select the mensuration wavelength of 440nm as total flavones, pharmacodynamics test proof Coryfolin be one of effective ingredient wherein again, so be reference substance mensuration content of total flavone with the Coryfolin.
The drafting of standard curve
Get psoralen reference substance 6.3mg, the accurate title, decide, and adds dissolve with methanol and become 25ml, and the accurate again 1ml that draws adds methanol and is diluted to 25ml, and accurate respectively then absorption diluent 1,2,3,4,5 adds methanol constant volume to 10ml, measures absorbance in the 243nm place.
Get Coryfolin reference substance 2.05mg, the accurate title, decide, and adds dissolve with methanol and become 50ml, respectively accurate then diluent 1,1.5,2,2.5, the 3ml of drawing adds the 1ml10%NaOH methanol solution, with methanol constant volume to 10ml, 50 ℃ of heating in water bath 2h are put coldly, measure absorbance in the 440nm place.
The regression equation of psoralen, Coryfolin is respectively y=0.1304x+0.0138 R as a result
2=0.9998; Y=0.0543x+0.0784 R
2=0.9996, respectively at 1.008-5.04 μ g.ml
-1And 4.10-12.30 μ g.ml
-1Between linear relationship good.
The precision examination
Get psoralen reference substance 2.52mg, the accurate title, decide, and adds dissolve with methanol and become 50ml, and the accurate again 5ml that draws adds methanol and is diluted to 10ml, measures absorbance, METHOD FOR CONTINUOUS DETERMINATION 5 times in the 243nm place.The accurate Coryfolin reference substance solution 1ml that draws adds the 1ml10%NaOH methanol solution, adds methanol constant volume again to 10ml, and 50 ℃ of heating in water bath 2h are put coldly, measure absorbance in 440nm place, METHOD FOR CONTINUOUS DETERMINATION 5 times, calculating RSD.RSD shows that all less than 1% instrument has higher precision as a result.
Examine stability
Psoralen reference substance, Coryfolin reference substance are measured absorbance at 243nm, 440nm place respectively in different time, calculating RSD, the stability of examination reference substance down with precision examination item.Get lot number and be 041201 sample, measure the absorbance of total coumarins and total flavones respectively in different time by the condition of drafting, calculate RSD, the examination stability of sample, psoralen reference substance, Coryfolin reference substance and sample are basicly stable at 48 hours internal absorbances as a result, and RSD is all less than 2%.
The repeatability examination
Get the sample (lot number: 041201) 5 parts of same lot number, measure total coumarins and content of total flavone respectively by method suggested, examine or check the repeatability of this method, the test of each time as a result content difference is little, RSD shows that all less than 2% total coumarins, content of total flavone assay method have repeatability preferably.
Recovery test
Get known content (lot number: 10 of slow releasing tablet 041201), porphyrize is got 0.25g, and accurate the title decides, accurate respectively psoralen reference substance solution, an amount of (three kinds of varying levels of Coryfolin reference substance solution of adding, each two parts), press the need testing solution preparation method and handle, carry out assay, calculate recovery rate, all between 95%-105%, the response rate is higher for the response rate of psoralen, Coryfolin as a result, shows that this method is feasible.
Two, the content of Fructus Psoraleae Coumarins and flavones ingredient in the HPLC method mensuration preparation
The preparation of need testing solution and reference substance solution
Get 10 of slow releasing tablet, porphyrize is got 0.5g, and accurate the title decides, and adds 20ml methanol supersound extraction 30min, filters, and adds methanol constant volume to 100ml, crosses 45 μ m filter membranes, gets subsequent filtrate, as need testing solution.
Precision takes by weighing psoralen 3.8mg, isopsoralen 3.3mg, isobavachin 1.7mg, Coryfolin 3.4mg, corylin 3.9mg, Corylifolinin 6.1mg, Fructus Psoraleae flavanone methyl ether 6.3mg, psoralidine 4.5mg, Bavachalcone 1.1mg respectively, add dissolve with methanol and become 25ml, as mixing reference substance solution.
Chromatographic condition
Chromatographic column: SUPELCO Discovery
C18 (25cm * 4.6mm, 5 μ m); Guard column: SUPELCOSupelguard
TMDiscovery
18 (2cm * 4.0mm, 5 μ m); Mobile phase: methanol: water=50: 50 (0-5min); 50: 50-100: 0 (5min-60min); F:1ml/min; λ: 320nm; T:30 ℃
The drafting of standard curve
Accurate respectively the absorption mixed reference substance solution 4 μ l, 8 μ l, 10 μ l, 16 μ l, 20 μ l inject high performance liquid chromatograph, measure peak area, the drawing standard curve, psoralen as a result, isopsoralen, isobavachin, Coryfolin, corylin, Corylifolinin, Fructus Psoraleae flavanone methyl ether, psoralidine, the regression equation of Bavachalcone is respectively y=26976x-87855, R=0.9999, y=20264x-61347, R=0.9999, y=9934.2x-26333, R=0.9999, y=13141x-38675, R=0.9999, y=7415.3x-23117, R=0.9999, y=27383x-151821, R=0.9999, y=11886x-70524, R=0.9999, y=18643x-106054, R=0.9999, y=27783x-103537, R=0.9999, the range of linearity is respectively 0.608～3.040 μ g, 0.528～2.640 μ g, 0.272～1.320 μ g, 0.544～2.720 μ g, 0.624～3.120 μ g, 0.976～4.880 μ g, 1.008～5.040 μ g, 0.720～3.600 μ g, 0.176 between～0.880 μ g.
The examination of precision
The accurate absorption mixed reference substance 10 μ l, injects high performance liquid chromatograph, measures peak area, and continuous sample introduction 5 times calculates RSD, the precision of examination instrument.The RSD of each reference substance shows that all less than 1% the precision of instrument is higher as a result.
Examine stability
To mix reference substance in different time sample introduction 10 μ l, measure peak area, calculate RSD, the stability of examination reference substance.Each reference substance is basicly stable in 48 hours as a result, and the peak area RSD that records in different time points is less than 2%.
The repeatability examination
Get same lot number sample (lot number: 041201) 5 parts, measure content by the assay method drafted, each time as a result tests that the content difference is little, RSD is less than 3%, this method has repeatability preferably.
The mensuration of the response rate
Get known content (lot number: 10 of slow releasing tablet 041201), porphyrize, get 0.25g, the accurate title, decide, accurate respectively psoralen reference substance solution, an amount of (three kinds of varying levels of Coryfolin reference substance solution of adding, each two parts), press the need testing solution preparation method and handle, carry out assay, calculate recovery rate, the response rate of psoralen, Coryfolin shows that this assay method is rationally feasible between 95%-105% as a result.
Three, the content of Radix Puerariae total flavones in the determined by ultraviolet spectrophotometry preparation
The preparation of need testing solution
Get 10 of slow releasing tablet, porphyrize is got 0.25g, and accurate the title decides, and adds 20ml chloroform supersound extraction 2 times, and each 10min discards chloroform solution, and residue adds dissolve with methanol, filters and is settled to 100ml, gets 0.1ml and is diluted to 100ml, as need testing solution.
The selection of absorbing wavelength
It is an amount of to get the puerarin reference substance, add dissolve with methanol, on ultraviolet-uisible spectrophotometer, scan, measure maximum absorption wavelength, simultaneously, need testing solution carries out UV scanning, and drug sustained-release tablet of the present invention has identical uv absorption with the puerarin reference substance, maximum absorption wavelength is 250nm, so select 250nm for measuring wavelength.
The drafting of standard curve
Precision takes by weighing puerarin reference substance 7.43mg, add dissolve with methanol and become 25ml, the accurate again 2.5ml that draws, add methanol and be diluted to the 25ml diluent, the above-mentioned diluent 1.0,1.5,2.0,2.5 of accurate respectively absorption, 3.0ml add methanol to scale in the 10ml measuring bottle, measure absorbance in the 250nm place, the regression equation of puerarin is as a result: A=0.0705C+0.1038 r=0.9999, concentration is at 2.972 and 8.916 μ g.ml
-1Between linear relationship good.
The precision examination
Precision takes by weighing puerarin reference substance 2.23mg, adds dissolve with methanol and becomes 50ml, and the accurate again 1ml that draws adds methanol constant volume to 25ml, measures absorbance in the 250nm place, and METHOD FOR CONTINUOUS DETERMINATION 5 times is calculated RSD, and RSD shows that less than 1% accuracy of instrument is higher as a result.
Examine stability
With the reference substance under the precision examination item, measure absorbance in the 250nm place at 0h, 4h, 8h, 12h, 24h respectively, calculate RSD.Get lot number and be 041201 sample, measure, measure absorbance, calculate RSD in different time by the condition of drafting.The result shows that the absorbance of puerarin reference substance and drug sustained-release tablet of the present invention is more stable in 24 hours, and RSD is less than 1%.
The repeatability examination
Get 5 parts in the sample (lot number 041201) of same lot number, carry out assay by the assay method of drafting, the content RSD of each time test shows that less than 2% this method has repeatability preferably.
Recovery test
Get drug sustained-release tablet of the present invention (lot number: 041201) 10, porphyrize, get 20mg, the accurate title, decide, accurate an amount of (the three kinds of varying levels of puerarin reference substance solution that add, each two parts), press the need testing solution preparation method and handle, carry out assay, calculate recovery rate, the response rate shows that this assay method has feasibility between 95%-105% as a result.
Four, the HPLC method is measured content of puerarin in the preparation
The preparation of need testing solution
Get 10 of slow releasing tablet, porphyrize is got 0.25g, and accurate the title decides, and adds the methanol ultrasonic dissolution, filters and is settled to 50ml, gets 1ml and is diluted to 100ml, as need testing solution.
Chromatographic condition
Chromatographic column: ODS post; Mobile phase: methanol: water=25: 75; F:1ml.min
-1λ: 250nm; T:30 ℃
The drafting of standard curve
Precision takes by weighing puerarin reference substance 7.43mg, add dissolve with methanol and become 25mL, the accurate 2ml that draws is in the 10ml measuring bottle, add methanol constant volume to scale, accurate respectively diluent 2,4,8,12,20, the 25 μ l injection high performance liquid chromatograph of drawing is measured by the chromatographic condition of drafting, and peak area and sample size are carried out linear regression, calculating regression equation is A=4E+06C-84328 r=0.99999, and puerarin reference substance sample size linear relationship between 0.11888-1.4860 μ g is good.
The precision examination
The accurate reference substance solution 10 μ l that draw under the standard curve drafting item inject high performance liquid chromatograph, and continuous sample introduction 5 times is measured by the chromatographic condition of drafting, and each time test peak area RSD shows that less than 1% accuracy of instrument is higher.
Examine stability
The accurate reference substance solution 10 μ l that draw respectively at 0h, 4h, 8h, 12h, 24h sample introduction, measure peak area, calculate RSD, and the result is basicly stable in the puerarin 24 hours.
The repeatability examination
Get 5 parts in the sample (lot number 041201) of same lot number, carry out assay by the assay method of drafting, each time result of the test RSD is less than 2% as a result, and this method has repeatability preferably.
Recovery test
Get slow releasing tablet (lot number: 041201) 10, porphyrize, get 50mg, the accurate title, decide, accurate an amount of (the three kinds of varying levels of puerarin reference substance solution that add, each two parts), press the need testing solution preparation method and handle, carry out assay, calculate recovery rate, the response rate of puerarin shows that this method is feasible between 98%-104% as a result.
The assay of drug sustained-release tablet of the present invention
Content, Radix Puerariae total flavones and the content of puerarin of Fructus Psoraleae coumarin and flavone in the test agent in measuring 3 batches by method suggested the results are shown in Table 10.
Table 10 drug sustained-release tablet assay of the present invention result
| Assay | Lot number | ??041201 | ??041202 | ????041203 | ||
| Fructus Psoraleae | UV method HPLC method | Total coumarins (mg/ sheet) total flavones (mg/ sheet) Coumarins (mg/ sheet) flavonoid (mg/ sheet) | ??26.22 ??26.92 ??14.18 ??15.99 | ??26.04 ??26.52 ??13.23 ??15.84 | ????25.57 ????26.41 ????13.18 ????15.15 | |
| Radix Puerariae | Total flavones (mg/ sheet) puerarin (mg/ sheet) | ??200.22 ??63.07 | ??205.49 ??68.56 | ????194.54 ????66.54 | ||
According to measurement result, in conjunction with factors such as the quality of medical material and extract and preparation process, decide every of this product and contain the amount of Fructus Psoraleae coumarin and flavone, survey with ultraviolet spectrophotometry, total coumarins must not be lower than 20mg in psoralen, and Fructus Psoraleae total flavones must not be lower than 20mg in Coryfolin; Survey with the HPLC method, every of Coumarins composition must not be lower than 10mg, every of bavachin must not be lower than 12mg; Every of Radix Puerariae total flavones must not be lower than 150mg, every of puerarin must not be lower than 50mg.
Below by pharmacodynamics, pharmacokinetics evidence beneficial effect of the present invention.
The 1 medicine material proportioning synergistic function test of the present invention of test example
Method: retinoic acid causes the osteoporosis rat animal model
Table 11 Fructus Psoraleae, Radix Puerariae are to the influence of osteoporosis rat bone density
| Group | Dosage (g/kg) | Number of animals (only) | Vertebra bone density (g/cm 2) | Femoral bmd (g/cm 2) |
| Fructus Psoraleae Radix Puerariae Fructus Psoraleae+Radix Puerariae model group normal control group | ??9 ??9 ??9+9 ??/ ??/ | ??6 ??6 ??6 ??5 ??6 | 0.1349±0.0112* 0.1373±0.0055* 0.1413±0.0081* 0.1089±0.0080* 0.1552±0.0144* | ??0.1370±0.0079* ??0.1334±0.0049* ??0.1401±0.0065* ??0.1099±0.0216* ??0.1501±0.0158* |
As can be seen from the above table, Fructus Psoraleae group, Radix Puerariae group and Fructus Psoraleae+Radix Puerariae group can both increase the bone density of retinoic acid osteoporosis rat significantly, and two medicines to share effect more obvious.
Cause the osteoporosis rat animal model by retinoic acid, Fructus Psoraleae, Radix Puerariae use separately or drug combination, all can increase rat vertebra bone density, femoral bmd significantly, but when Fructus Psoraleae and Radix Puerariae consumption were each 9g/kg, the effect of its bone density improving was better than independent utilization.
Test example 2 drug sustained-release tablet cumulative in vitro releases of the present invention
The selection of release medium and assay method:
This product active substance is Radix Puerariae total flavones, Fructus Psoraleae Coumarins and flavones ingredient, for acidulous material or lactone structure is arranged, in water and acid medium in dissolubility little, do not reach sink conditions (dropping into the 10%-20% that dose must not be crossed dissolubility), and can dissolve preferably in the solution of meta-alkalescence slightly, so the phosphate buffer of selecting PH=7.4 is as dissolution medium.
In experiment, compared commentaries on classics basket method and slurry method, good with the repeatability of changeing basket method mensuration release than the slurry method, during with the slurry method, at the bottom of tablet easily sticked at glass, mixing effect was bad, and the tablet motion is irregular, cause the measurement result repeatability relatively poor, change the release that the basket method is measured this product so select for use.
The mensuration of sample release
Get test agent in 3 batches of known content, 6 every batch, press two dissolution method first methods of the Pharmacopoeia of the People's Republic of China (changeing the basket method) operation, respectively at 1,2,3,4,6,8,10,12 hour sampling 5ml, add dilute hydrochloric acid and transfer pH5-6, water bath method, residue add 10ml chloroform supersound extraction 2 times, each 10min, collect chloroform solution, residue adds dissolve with methanol and is settled to 25ml, measures absorbance in the 250nm place, calculates the cumulative release degree of Radix Puerariae total flavones.Chloroform solution evaporate to dryness, residue add dissolve with methanol and are settled to 25ml, measure the cumulative release degree of total coumarins in the 243nm place, the results are shown in Table 12,13.
Radix Puerariae total flavones accumulation stripping percentage rate measurement result in table 12 drug sustained-release tablet of the present invention
| Lot number | Sequence number | Cumulative release percentage rate (%) | |||||||
| ?1h | ?2h | ??3h | ??4h | 6h | ??8h | ??10h | ??12h | ||
| ??0 ??4 ??1 ??2 ??0 ??1 | 123456 is average | ?16.32 ?18.24 ?20.31 ?21.08 ?18.99 ?19.37 ?19.05 | ?27.31 ?29.64 ?32.18 ?35.47 ?31.85 ?32.11 ?31.42 | ??40.63 ??41.85 ??45.32 ??47.33 ??43.24 ??38.72 ??42.84 | ??53.15 ??55.34 ??57.28 ??56.19 ??50.44 ??49.62 ??53.67 | 68.14 70.51 72.20 69.84 64.97 65.83 68.58 | ??80.43 ??81.62 ??80.99 ??77.47 ??74.61 ??76.38 ??78.58 | ??87.61 ??89.27 ??86.45 ??83.11 ??84.20 ??81.04 ??85.28 | ??92.49 ??94.61 ??92.10 ??91.88 ??90.74 ??89.23 ??91.84 |
| ??0 ??4 ??1 ??2 ??0 ??2 | 123456 is average | ?18.54 ?18.99 ?19.28 ?20.10 ?18.46 ?17.99 ?18.89 | ?27.39 ?32.41 ?33.13 ?32.87 ?30.54 ?28.66 ?30.83 | ??38.18 ??43.73 ??46.82 ??44.31 ??43.86 ??37.25 ??45.36 | ??53.43 ??56.24 ??59.31 ??55.47 ??56.33 ??50.86 ??56.27 | 68.55 71.29 70.34 67.27 69.44 65.78 68.77 | ??81.53 ??83.46 ??80.91 ??76.30 ??79.24 ??77.01 ??79.74 | ??88.00 ??89.71 ??87.16 ??87.02 ??83.11 ??82.07 ??86.18 | ??95.37 ??93.44 ??95.19 ??92.77 ??89.87 ??87.63 ??92.38 |
| ??0 ??4 ??1 ??2 ??0 ??3 | 123456 is average | ?15.29 ?18.16 ?20.48 ?16.01 ?19.07 ?15.54 ?17.43 | ?32.27 ?33.55 ?36.32 ?33.31 ?35.33 ?31.87 ?33.78 | ??43.54 ??46.83 ??45.27 ??50.54 ??47.36 ??42.75 ??46.05 | ??56.78 ??59.36 ??55.72 ??54.37 ??60.29 ??55.50 ??57.00 | 73.32 76.48 71.11 74.24 77.47 69.49 73.69 | ??85.43 ??83.64 ??81.25 ??86.77 ??85.81 ??80.34 ??83.87 | ??92.90 ??86.26 ??87.13 ??90.49 ??86.68 ??86.22 ??88.28 | ??95.63 ??92.81 ??90.26 ??96.03 ??97.24 ??90.71 ??93.78 |
Fructus Psoraleae total coumarins accumulation stripping percentage rate measurement result in table 13 drug sustained-release tablet of the present invention
| Lot number | Sequence number | Accumulation stripping percentage rate (%) | |||||||
| ??1h | ?2h | ??3h | ??4h | ??6h | ??8h | ??10h | ??12h | ||
| ?0 ?4 ?1 ?2 ?0 ?1 | 123456 is average | ??10.83 ??12.67 ??14.32 ??10.19 ??9.87 ??13.46 ??11.89 | ?22.38 ?24.63 ?28.71 ?26.15 ?24.33 ?27.30 ?25.58 | ??37.41 ??39.20 ??41.37 ??38.88 ??37.74 ??40.26 ??39.14 | ??46.38 ??50.01 ??52.64 ??49.50 ??45.97 ??48.81 ??48.89 | ??59.07 ??63.43 ??63.58 ??62.00 ??59.67 ??60.84 ??61.43 | ??70.31 ??72.47 ??73.01 ??70.18 ??67.50 ??69.56 ??70.51 | ??79.93 ??77.80 ??81.34 ??79.52 ??75.06 ??78.77 ??78.74 | ??88.37 ??90.65 ??89.48 ??87.60 ??88.94 ??85.71 ??88.46 |
| ?0 ?4 ?1 ?2 ?0 ?2 | 123456 is average | ??12.41 ??11.08 ??13.48 ??10.03 ??11.87 ??9.92 ??11.46 | ?26.35 ?24.68 ?29.06 ?25.49 ?27.83 ?23.04 ?26.08 | ??41.75 ??39.02 ??45.37 ??38.44 ??40.10 ??37.63 ??40.38 | ??51.59 ??50.79 ??54.25 ??48.04 ??49.23 ??46.85 ??50.13 | ??62.54 ??64.57 ??63.58 ??61.24 ??63.03 ??59.37 ??63.06 | ??74.49 ??71.82 ??72.64 ??69.33 ??71.26 ??68.97 ??71.42 | ??84.61 ??80.76 ??83.14 ??80.51 ??83.07 ??78.59 ??81.78 | ??92.77 ??89.31 ??89.31 ??86.45 ??88.27 ??85.46 ??88.60 |
| ?0 ?4 ?1 ?2 ?0 ?3 | 123456 is average | ??13.41 ??13.87 ??14.25 ??12.16 ??13.99 ??15.37 ??13.84 | ?22.18 ?25.72 ?24.11 ?20.76 ?22.85 ?23.17 ?23.13 | ??37.68 ??39.55 ??37.40 ??35.51 ??38.41 ??36.38 ??37.49 | ??51.54 ??53.29 ??50.18 ??48.74 ??49.33 ??50.14 ??50.54 | ??61.07 ??65.36 ??62.11 ??60.43 ??62.38 ??60.97 ??62.05 | ??73.63 ??75.42 ??72.04 ??70.84 ??73.68 ??69.61 ??72.54 | ??83.05 ??82.73 ??83.31 ??76.15 ??80.07 ??78.64 ??80.65 | ??89.43 ??90.74 ??89.67 ??86.35 ??92.48 ??90.17 ??89.80 |
Drug sustained-release tablet release rule of the present invention
The mathematical model that is used for drug release at present has zero level model, one-level model, Higuchi model and Ritger-Pappas model, with above-mentioned drug release kinetics model the dispose procedure of drug sustained-release tablet of the present invention is carried out match, inquire into the release mechanism of slow releasing tablet.
The average accumulated release of test agent in three batches is carried out match with the time with different release models, the results are shown in Table 14,15.
Table 14 Radix Puerariae total flavones release model fitting result
| Lot number | Model of fit | Fit equation | Correlation coefficient |
| ??041201 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ?F(t)＝0.0649t+0.2161 ?Ln[1-F(t)]＝-0.2033t+0.0391 ?F(t)＝3.2672t 1/2+0.3317 ?Ln?F(t)＝0.6395Lnt-1.5895 ?Ln{-Ln[1-F(t)]}＝0.994Lnt-1.6238 | ??R 2＝0.9423 ??R 2＝0.9951 ??R 2＝0.9902 ??R 2＝0.9874 ??R 2＝0.9971 |
| ??041202 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ?F(t)＝0.065t+0.2243 ?Ln[1-F(t)]＝-0.2091t+0.037 ?F(t)＝3.233t 1/2+0.3229 ?LnF(t)＝0.6445Lnt-1.5833 ?Ln{-Ln[1-F(t)]}＝1.0082Lnt-1.6176 | ??R 2＝0.9308 ??R 2＝0.9952 ??R 2＝0.9852 ??R 2＝0.9797 ??R 2＝0.9970 |
| ??041203 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ?F(t)＝0.0669t+0.2327 ?Ln[1-F(t)]＝-0.2309t+0.0592 ?F(t)＝3.0938t 1/2+0.3463 ?LnF(t)＝0.6694Lnt-1.5954 ?Ln{-Ln[1-F(t)]}＝1.0667Lnt-1.643 | ??R 2＝0.9115 ??R 2＝0.9969 ??R 2＝0.9763 ??R 2＝0.9683 ??R 2＝0.9992 |
Table 15 Fructus Psoraleae total coumarins release model fitting result
| Lot number | Model of fit | Fit equation | Correlation coefficient |
| ??041201 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ??F(t)＝0.0654t+0.1549 ??Ln[1-F(t)]＝-0.1718t+0.054 ??F(t)＝3.2505t 1/2+0.5309 ??LnF(t)＝0.7765Lnt-1.942 ??Ln{-Ln[1-F(t)]}＝1.0886Lnt-1.989 | ??R 2＝0.9459 ??R 2＝0.9849 ??R 2＝0.9917 ??R 2＝0.9668 ??R 2＝0.9940 |
| ??041202 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ??F(t)＝0.0664t+0.1595 ??Ln[1-F(t)]＝-0.1783t+0.0553 ??F(t)＝3.1835t 1/2+0.5336 ??LnF(t)＝0.7909Lnt-1.9487 ??Ln{-Ln[1-F(t)]}＝1.1149Lnt-1.9992 | ??R 2＝0.9377 ??R 2＝0.9927 ??R 2＝0.9889 ??R 2＝0.9589 ??R 2＝0.9931 |
| ??041203 | Zero level one-level Higuchi Ritger-Pappas Weibull (weibull) | ??F(t)＝0.0675t+0.1496 ??Ln[1-F(t)]＝-0.1843t+0.0881 ??F(t)＝3.1529t 1/2+0.5615 ??LnF(t)＝0.7614Lnt-1.9048 ??Ln{-Ln[1-F(t)]}＝1.0907Lnt-1.9637 | ??R 2＝0.9492 ??R 2＝0.9853 ??R 2＝0.9900 ??R 2＝0.9784 ??R 2＝0.9932 |
Correlation coefficient adopts above each model as can be seen from table 14, table 15, and all there are dependency preferably release and time, and wherein the match of Weibull (weibull) model is best, shows that the external release rule of drug sustained-release tablet of the present invention meets Weibull model.
Test example 3 drug sustained-release tablets of the present invention are in the intravital pharmacokinetics test of Canis familiaris L.
Experiment material and instrument
Drug sustained-release tablet of the present invention, medicine conventional tablet of the present invention (being self-control), methanol (chromatographically pure), water (deionized water), other reagent of acetonitrile (chromatographically pure) are analytical pure.
6 of Beagle dogs, body weight 10 ± 0.1Kg; Male and female half and half are provided by the Chongqing Institute of Chinese Medicine Experimental Animal Center.
Waters 2695 separations module high performance liquid chromatographs, Waters 2996 photodiode arraydetector detectors, Waters empowder chem workstation; TGL-16C table model high speed centrifuge (Anting Scientific Instrument Factory, Shanghai); XW-80A vortex mixer (Qingpu, Shanghai Luxi instrument plant).
The foundation of determination of plasma concentration method
The present invention has adopted the HPLC chromatography, can be under identical conditions separately with composition to be measured, and good separating effect.Chromatographic condition is: chromatographic column: SUPELCO Discovery
The C18 chromatographic column; SUPELCO Supelguard
TMDiscovery
18 guard columns; Mobile phase: gradient elution 0min acetonitrile: water (5: 95), 20min acetonitrile: water (15: 85), 40min acetonitrile: water (10: 60), 60min acetonitrile: water (70: 30) 80min acetonitrile: water (5: 95); Flow velocity: 1ml/min, column temperature: 30 ℃, detect wavelength: 250nm.
Puerarin is the main effective ingredient in the Radix Puerariae flavone, the content height, psoralen, isopsoralen are the effective ingredient in the Fructus Psoraleae, content is also higher relatively, so selecting puerarin, psoralen, isopsoralen is representative, examines or check it in the intravital kinetics of dog.And to its range of linearity, withinday precision, day to day precision, the response rate are examined or check, to prove the feasibility of this assay method.
The preparation of blank blood serum sample and pastille blood serum sample
Accurate blank serum 0.5ml, adding methanol 3.5ml, vortex vibration 1min, centrifugal 10min (10000r/min), the supernatant N of drawing
2Dry up, residue adds the 1ml dissolve with methanol, and centrifugal 10min (10000r/min) gets the supernatant sample introduction, measures chromatogram.
Accurate draw blank serum 0.5ml, it is an amount of to add puerarin reference substance, psoralen reference substance, isopsoralen reference substance, adds methanol 3.5ml again, operates by blank blood serum sample preparation method in accordance with the law, gets the supernatant sample introduction, measures chromatogram.
The mensuration of minimum detectable level
3 times (S/N=3) with machine noise are the boundary, and the lowest detectable limit of puerarin, psoralen, isopsoralen is respectively 0.05ng, 0.1ng, 0.1ng.
The drafting of standard curve
Get 5 parts of blank serum 0.5ml, add puerarin reference substance solution (336 μ g/ml), psoralen reference substance solution (2.84 μ g/ml), isopsoralen reference substance solution (2.6 μ g/ml) each 100 μ l, 80 μ l, 60 μ l, 40 μ l, 20 μ l respectively, add 3.5ml methanol again, eddy current mixing 1min, high speed centrifugation (10000 change) 10min, incline and get supernatant, 50 ℃ of N
2Dry up, add the 1ml dissolve with methanol, high speed centrifugation (10000r/min) 10min gets supernatant, the accurate respectively 10 μ l of absorption inject high performance liquid chromatogram, measure peak area, are vertical coordinate with peak area integrated value Y, sample size X is an abscissa, and the drawing standard curve calculates regression equation.
The standard curve determination result of table 16 reference substance in serum
| The puerarin reference substance | The psoralen reference substance | The isopsoralen reference substance | |||
| Sample size (μ g) | Peak area | Sample size (ng) | Peak area | Sample size (ng) | Peak area |
| 0.336 0.2688 0.2016 0.1344 0.0672 | ????807601 ????644372 ????485272 ????328523 ????156938 | ??2.84 ??2.272 ??1.704 ??1.136 ??0.568 | ????18583 ????14680 ????10778 ????7619 ????4088 | ??2.6 ??2.08 ??1.56 ??1.04 ??0.52 | ????15188 ????12226 ????8963 ????5999 ????3285 |
Can calculate from table 16, the regression equation of puerarin is Y=2405393.75X-611.3 (R
2=9998), good in 0.0672 μ g-0.336 μ g scope internal linear relation; The regression equation of psoralen is Y=6347X+334.3 (R
2=0.9983), good in 0.568-2.84ng scope internal linear relation; The regression equation of isopsoralen is Y=5775.6X+122.3 (R
2=0.9991), good in 0.52-2.6ng scope internal linear relation.
The precision test
The withinday precision of puerarin, psoralen, isopsoralen all is smaller than 5%, and day to day precision show that instrument has higher precision, and sample is basicly stable in two days all less than 10%.
Recovery test
Get blank serum 0.5ml, add not commensurability puerarin, psoralen, isopsoralen reference substance solution respectively, the vortex mix homogeneously adds methanol 3.5ml respectively, vortex vibration 1min, centrifugal 10min (10000r/min), supernatant N
2Dry up, residue adds the 1ml dissolve with methanol, and centrifugal 10min (10000r/min) gets the supernatant sample introduction.
The response rate of each reference substance shows that this method is rationally feasible in claimed range.
In vivo assay Cells and interpretation of result
Get 6 of Beagle dogs, be divided into 2 groups at random, 3 every group, fasting 12h before the test, irritate respectively stomach give drug sustained-release tablet of the present invention and ordinary tablet (1/kg), 10min after administration, 20min, 0.5h, 1h, 2h, 3h, 4h, 5h, 6h, 8h, 10h gets blood from femoral vein, leaves standstill behind the 2h centrifugally, and it is standby to get upper serum.Measure blood drug level by method suggested and condition, adopt the data of 3P87 pharmacokinetics program his-and-hers watches 17～19 to handle, select the chamber model, calculate goodness of fit parameter.
In table 17 medicine conventional tablet of the present invention and the slow releasing tablet puerarin on average through the time blood drug level goodness of fit parameter
| Group | ??No ??Comp. | ????WT | ????R ?Squares | ??Goodness ??of?Fit | ??Max.Error ??C-CL | ????Max. ????Error％ | ??AIC | ?Run?Test |
| Mend sclerotin ordinary tablet group | ??1 ??2 ??3 ??1 ??2 ??3 ??1 ??2 ??3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ?0.9894 ?0.9898 ?0.9979 ?0.9971 ?0.9993 ?0.9994 ?0.9993 ?0.9998 ?0.9997 | ??0.2590 ??0.3012 ??0.1778 ??0.1353 ??0.0768 ??0.0977 ??0.0672 ??0.0455 ??0.0613 | ??0.4830 ??0.4805 ??0.1876 ??0.4716 ??0.2044 ??0.2004 ??0.5517 ??0.2051 ??0.2233 | ????18.23 ????18.10 ????18.74 ????12.54 ????5.44 ????5.33 ????11.02 ????5.45 ????5.94 | ??-0.3157 ??3.3008 ??-9.9098 ??-14.6064 ??-26.7514 ??-23.0864 ??-29.9965 ??-38.2550 ??-33.3255 | ????4 ????3 ????6 ????4 ????5 ????5 ????6 ????5 ????5 |
| Mend sclerotin slow releasing tablet group | ??1 ??2 ??3 ??1 ??2 ??3 ??1 ??2 ??3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ?0.8980 ?0.9140 ?0.8890 ?0.9618 ?0.9572 ?0.9869 ?0.9811 ?0.9900 ?0.9819 | ??0.5578 ??0.5810 ??0.7808 ??0.3415 ??0.4096 ??0.2686 ??0.2399 ??0.2345 ??0.2667 | ??1.1454 ??0.9677 ??0.7514 ??1.4120 ??1.0990 ??0.7544 ??1.8836 ??1.1922 ??1.8508 | ????76.84 ????75.77 ????73.94 ????52.81 ????64.87 ????42.20 ????38.84 ????29.59 ????38.16 | ??21.3861 ??23.1786 ??30.4885 ??8.6302 ??14.0889 ??2.7402 ??-0.5544 ??-0.7852 ??2.9302 | ????3 ????3 ????3 ????3 ????3 ????3 ????3 ????4 ????5 |
In table 18 medicine conventional tablet of the present invention and the slow releasing tablet psoralen on average through the time blood drug level goodness of fit parameter
| Group | ??No ??Comp. | ????WT | ????R ??Squares | ????Goodness ????of?Fit | ??Max.Error ??C-CL | ??Max. ??Error％ | ??AIC | ??Run?Test |
| Mend sclerotin ordinary tablet group | ??1 ??2 ??3 ??1 ??2 ??3 ??1 ??2 ??3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ??0.8726 ??0.8715 ??0.9324 ??0.1035 ??-0.4067 ??0.4414 ??-6.1191 ??-11.9217 ??-12.1452 | ????3.1841 ????0.0378 ????0.0354 ????0.0845 ????0.1252 ????0.1018 ????0.2380 ????0.3464 ????0.4940 | ??0.0662 ??0.0664 ??0.0376 ??0.0784 ??0.0444 ??0.0563 ??0.0942 ??0.1306 ??0.0673 | ??74.18 ??70.96 ??81.94 ??36.15 ??87.22 ??43.71 ??37.37 ??51.81 ??58.63 | ??-46.4288 ??-42.3358 ??-45.3937 ??-24.9664 ??-16.0108 ??-22.1694 ??-2.1719 ??6.3834 ??12.5720 | ????4 ????4 ????4 ????4 ????4 ????4 ????4 ????3 ????4 |
| Mend sclerotin slow releasing tablet group | ??1 ??2 ??3 ??1 ??2 ??3 ??1 ??2 ??3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ??0.8354 ??0.7999 ??0.8265 ??-0.5230 ??-0.4928 ??0.0389 ??-15.0553 ??-13.3191 ??-11.3269 | ????0.0303 ????0.0386 ????0.0441 ????0.0923 ????0.1055 ????0.1037 ????0.2997 ????0.3268 ????0.3321 | ??0.0411 ??0.0526 ??0.0525 ??0.0617 ??0.0606 ??0.0438 ??0.0883 ??0.0774 ??0.0729 | ??78.32 ??56.75 ??80.77 ??40.29 ??46.66 ??57.95 ??43.30 ??37.94 ??35.93 | ??-50.9311 ??-44.5886 ??-42.2988 ??-24.2341 ??-20.4745 ??-21.7584 ??4.0304 ??6.6570 ??6.8592 | ????3 ????3 ????3 ????3 ????3 ????3 ????3 ????3 ????3 |
In the table 19 medicine conventional tablet of the present invention isopsoralen on average through the time blood drug level goodness of fit parameter
| Group | ???No ???Comp. | ????WT | ????R ??Squares | ??Goodness ??of?Fit | ??Max.Error ??C-CL | ??Max. ??Error％ | ?????AIC | ?Run?Test |
| Mend sclerotin ordinary tablet group | ????1 ????2 ????3 ????1 ????2 ????3 ????1 ????2 ????3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ??0.7089 ??0.7345 ??0.7823 ??-1.7966 ??-1.7098 ??-1.2399 ??-27.2595 ??-27.9374 ??-23.1917 | ??0.0306 ??0.0346 ??0.0405 ??0.0950 ??0.1106 ??0.1298 ??0.3019 ??0.3615 ??0.4267 | ??0.0621 ??0.0501 ??0.0365 ??0.0718 ??0.0685 ??0.0629 ??0.0853 ??0.0834 ??0.0749 | ??78.59 ??76.89 ??87.73 ??44.32 ??47.43 ??49.43 ??52.65 ??51.50 ??46.25 | ??-47.2749 ??-44.2863 ??-42.4685 ??-22.3869 ??-18.7339 ??-16.8282 ??3.0565 ??7.3169 ??9.3466 | ????4 ????3 ????4 ????4 ????4 ????4 ????4 ????3 ????4 |
| Mend sclerotin slow releasing tablet group | ????1 ????2 ????3 ????1 ????2 ????3 ????1 ????2 ????3 | ????1 ????1 ????1 ????1/C ????1/C ????1/C ????1/C/C ????1/C/C ????1/C/C | ??0.7502 ??0.7945 ??0.8384 ??-1.5024 ??-1.4359 ??-2.6069 ??-35.9333 ??-31.9222 ??-17.0896 | ??0.0245 ??0.0263 ??0.0301 ??0.0776 ??0.0906 ??0.1424 ??0.2983 ??0.3332 ??0.3189 | ??0.0474 ??0.0370 ??0.0304 ??0.0504 ??0.0493 ??0.0351 ??0.0667 ??0.0592 ??0.0483 | ??59.54 ??58.84 ??100 ??50.93 ??51.46 ??100 ??48.36 ??42.89 ??35.00 | ??-52.1671 ??-50.3139 ??-48.9577 ??-26.8173 ??-23.1138 ??-14.7959 ??2.7930 ??5.5284 ??2.9415 | ????4 ????3 ????4 ????3 ????3 ????4 ????3 ????3 ????3 |
According to the minimum principle of AIC, from table 17～19 as can be seen, puerarin is two-compartment model in ordinary tablet and slow releasing tablet, and weight is 1/C/C, and psoralen, isopsoralen are one-compartment model in ordinary tablet and slow releasing tablet, and weight is 1.Respectively organize pharmacokinetic parameters according to chamber model and weight calculation, the results are shown in Table 20～23.
The pharmacokinetic parameters of puerarin in table 20 medicine conventional tablet of the present invention and the slow releasing tablet
| Parameter | ?????????????Common-release?tablet | ????????????Sustained-release?tablet | ||||||
| ??No.1 | No.2 | ?No.3 | ?mean | ?No.1 | ?No.2 | ????No.3 | ??mean | |
| A(μg/mL) Alpha(l/h) B(μg/mL) beta(l/h) Ka(l/h) T 1/2alpha(h) T 1/2beta(h) T 1/2Ka(h) AUC(μg/mL)*h Tpeak(h) Cmax(μg/mL) CL/F(s)mg/h/μg/mL | ??6.2908 ??5.1381 ??8.0368 ??0.3441 ??5.8856 ??0.1349 ??2.0143 ??0.1178 ??22.1448 ??0.4814 ??6.4978 ??189.66 | 5.9506 0.6576 3.2199 0.1761 6.5445 1.0540 3.9353 0.1059 25.9283 0.4409 6.9205 161.98 | ?6.5942 ?0.2293 ?0.0619 ?0.2104 ?2.0293 ?3.0224 ?3.2949 ?0.3416 ?25.7677 ?1.2167 ?4.4730 ?162.99 | ?6.2785 ?2.0083 ?3.7729 ?0.2435 ?4.8198 ?1.4038 ?3.0815 ?0.1884 ?24.6136 ?0.7130 ?5.9638 ?171.54 | ?2.0857 ?0.1615 ?5.2853 ?0.1297 ?0.3832 ?4.2915 ?5.3448 ?1.8090 ?34.4309 ?4.1854 ?2.6457 ?121.98 | ?2.5356 ?0.4173 ?14.4379 ?0.1742 ?0.3444 ?1.6612 ?3.9795 ?2.0129 ?39.6760 ?4.1188 ?3.3909 ?105.85 | ????9.6448 ????0.2111 ????8.4829 ????0.1597 ????0.3172 ????3.2829 ????4.3412 ????2.1852 ????41.6582 ????4.1373 ????3.5284 ????100.82 | ??4.7554 ??0.2633 ??9.4020 ??0.1545 ??0.3483 ??3.0789 ??4.5552 ??2.0024 ??38.5884 ??4.1472 ??3.1883 ??109.55 |
The pharmacokinetic parameters of psoralen in table 21 medicine conventional tablet of the present invention and the slow releasing tablet
| ??Parameter | ??????????????Common-release?tablet | ??????????Sustained-release?tablet | ||||||
| ??No.1 | ??No.2 | ??No.3 | ??mean | ?No.1 | ?No.2 | ??No.3 | ??mean | |
| ??A(μg/mL) ??Ke(l/h) ??Ka(l/h) ??T 1/2Ke(h) ??T 1/2Ka(h) ??Tpeak(h) ??Cmax(μg/mL) ??AUC(μg/mL)*h ??CL/F(s)mg/h/μg/mL | ??0.4436 ??0.3427 ??0.9284 ??2.0230 ??0.7466 ??1.7016 ??0.1562 ??0.8168 ??5141.99 | ??0.8443 ??0.4022 ??0.7592 ??1.7236 ??0.9130 ??1.7797 ??0.1941 ??0.9873 ??4254.09 | ??0.8079 ??0.3040 ??0.6207 ??2.2803 ??1.1167 ??2.2540 ??0.2078 ??1.3562 ??3096.91 | ??0.6986 ??0.3496 ??0.7694 ??2.0089 ??0.9254 ??1.9118 ??0.1860 ??1.0534 ??4164.33 | ?0.6868 ?0.1885 ?0.3238 ?3.6764 ?2.1404 ?3.9981 ?0.1350 ?1.5220 ?2759.53 | ?0.6376 ?0.1797 ?0.3467 ?3.8580 ?1.9993 ?3.9355 ?0.1515 ?1.7097 ?2456.60 | ??0.7352 ??0.1727 ??0.3326 ??4.0146 ??2.0839 ??4.0991 ??0.1742 ??2.0477 ??2051.08 | ??0.6865 ??0.1803 ??0.3344 ??3.8497 ??2.0745 ??4.0109 ??0.1536 ??1.7598 ??2422.40 |
The pharmacokinetic parameters of isopsoralen in table 22 medicine conventional tablet of the present invention and the slow releasing tablet
| Parameter | ????????????Common-release?tablet | ?????????Sustained-release?tablet | ||||||
| ??No.1 | ?No.2 | ???No.3 | ??mean | ??No.1 | ??No.2 | ?No.3 | ??mean | |
| A(μg/mL) Ke(l/h) Ka(l/h) T 1/2Ke(h) T 1/2Ka(h) Tpeak(h) Cmax(μg/mL) AUC(μg/mL)*h CL/F(s)mg/h/μg/mL | ?0.4842 ?0.3754 ?0.6521 ?1.8463 ?1.0629 ?1.9956 ?0.0971 ?0.5473 ?7674.61 | ?0.6165 ?0.3606 ?0.6126 ?1.9220 ?1.1315 ?2.1029 ?0.1188 ?0.7030 ?5973.97 | ???0.2243 ???0.3126 ???1.3943 ???2.2176 ???0.4971 ???1.3823 ???0.1130 ???0.5568 ???774543.54 | ??0.4417 ??0.3495 ??0.8863 ??1.9953 ??0.8972 ??1.8269 ??0.1096 ??0.6024 ??7064.04 | ??0.3074 ??0.1736 ??0.3608 ??3.9919 ??1.9213 ??3.9077 ??0.0809 ??0.9182 ??4574.28 | ??0.4114 ??0.1791 ??0.3391 ??3.8693 ??2.0442 ??3.9895 ??0.0950 ??1.0833 ??3877.12 | ?0.4443 ?0.1801 ?0.3358 ?3.8482 ?2.0641 ?4.0010 ?0.1002 ?1.1436 ?3672.49 | ??0.3877 ??0.1776 ??0.3452 ??3.9031 ??2.0098 ??3.9661 ??0.0920 ??1.0484 ??4041.29 |
The main pharmacokinetic parameters of table 23 medicine conventional tablet of the present invention and slow releasing tablet relatively
| ?component | ??Parameter | ????Common-release ????tablet | ???Sustained-release ???tablet | ????T?test | ????P |
| ?puerariae ?psoralen ?isopsoralen | ??AUC(0→∞) ??Cmax ??Tmax ??MRT ??AUC ??Cmax ??Tmax ??MRT ??AUC ??Cmax ??Tmax ??MRT | ????24.6136 ????5.9638 ????0.7130 ????3.8623 ????1.0534 ????0.1860 ????1.9118 ????5.6200 ????0.6024 ????0.1096 ????1.8269 ????5.6288 | ????41.6582 ????3.5284 ????4.1373 ????9.2577 ????1.7598 ????0.1536 ????4.0109 ????9.0813 ????1.0484 ????0.0920 ????3.9661 ????9.2411 | ????5.624 ????3.4540 ????13.579 ????3.6110 ????3.1911 ????1.6941 ????11.7234 ????3.9637 ????5.3012 ????2.0280 ????9.4494 ????3.9637 | ????＜0.01 ????＜0.05 ????＜0.01 ????＜0.05 ????＜0.05 ????＞0.05 ????＜0.01 ????＜0.01 ????＜0.01 ????＞0.05 ????＜0.01 ????＜0.01 |
t
0.05＝2.1318，t
0.01＝3.7470
As can be seen from Table 23, the main pharmacokinetic parameters of drug sustained-release tablet of the present invention and ordinary tablet has significant difference, show slow releasing tablet can be in the dog body long period keep higher blood drug level.
Drug sustained-release tablet of the present invention inside and outside dependency
The inside and outside dependency is the function of absorption fraction in release in vitro parameter and the body, and the purpose of inside and outside correlation research is to set up a quality control index of we can say the quality standard of open-birth thing availability and being used as preparation when producing in batches.
Discharging percentage rate (F) with cumulative in vitro is independent variable, with absorption fraction (f) in the t body constantly is dependent variable, carry out linear regression with method of least square, by comparing the marginal value of correlation coefficient and r, the examination cumulative in vitro discharges the dependency of absorption fraction in percentage rate and the body.
Puerarin is a two-compartment model in the drug sustained-release tablet of the present invention, with formula F a=(Ct+K ∫
0Ctdt+X (p)/V
C)/kK ∫
0 ∞Ctdt*100% calculates dependency, the results are shown in Table 24.Psoralen is an one-compartment model, with formula F a=(Ct+K ∫
0Ctdt)/kK ∫
0 ∞Ctdt*100% calculates dependency, the results are shown in Table 25.
The interior absorption fraction f (%) of puerarin body discharges percentage rate F (%) with cumulative in vitro in table 24 drug sustained-release tablet of the present invention
| Time (h) | ????Ct | ????K∫ 0 Ctdt | ????X(p)/Vc | ??Ct+K∫ 0 Ctdt+X(p)/V C | ????f％ | ????F％ |
| ????1 ????2 ????3 ????4 ????6 ????8 ????10 ????12 | ????1.68 ????3.09 ????4.11 ????4.98 ????3.74 ????2.41 ????1.66 ????1.24 | ????0.8570 ????2.9174 ????5.6147 ????8.5729 ????14.4067 ????19.4506 ????23.4644 ????26.5105 | ????5.9162E-5 ????8.8371E-5 ????1.0997E-4 ????1.2149E-4 ????1.2509E-4 ????1.1295E-4 ????9.6142E-5 ????7.8572E-5 | ??2.1019 ??4.5263 ??6.8741 ??9.2004 ??10.8324 ??11.9855 ??13.2115 ??14.2911 | ????12.52 ????26.95 ????40.93 ????54.78 ????64.50 ????71.37 ????78.67 ????85.10 | ????19.05 ????31.42 ????42.84 ????53.67 ????68.58 ????78.58 ????85.28 ????91.84 |
The interior absorption fraction f (%) of psoralen body discharges percentage rate F (%) with cumulative in vitro in table 25 drug sustained-release tablet of the present invention
| Time (h) | ????Ct | ??K∫ 0 Ctdt | ?Ct+K∫ 0 Ctdt | ????f％ | ????F％ |
| ????1 ????2 ????3 ????44 ????6 ????8 ????10 ????12 | ????0.045 ????0.098 ????0.155 ????0.193 ????0.178 ????0.108 ????0.081 ????0.065 | ????0.0377 ????0.1277 ????0.2442 ????0.3708 ????0.6197 ????0.8361 ????1.0116 ????1.1487 | ????0.0618 ????0.1552 ????0.2643 ????0.3590 ????0.4554 ????0.4823 ????0.5339 ????0.8243 | ????8.82 ????22.08 ????37.61 ????51.08 ????64.81 ????68.63 ????75.97 ????82.43 | ????11.89 ????25.58 ????39.14 ????48.89 ????61.43 ????70.51 ????78.74 ????88.46 |
The inside and outside dependency equation that can calculate Radix Puerariae flavone from table 24 is: Y=0.0097X-0.0273, R
2=0.9888; The dependency equation that can calculate coumarin from table 25 is: Y=99.758X+1.7758, R
2=0.9867, the result shows that interior the absorption with release in vitro of the body of Radix Puerariae flavone and Fructus Psoraleae coumarin has dependency preferably, shows that release in vitro degree method is rationally feasible, can be used for replacing in vivo test to carry out the quality control of product.
Claims (13)
1, a kind of pharmaceutical composition for the treatment of osteoporosis is characterized in that: it is the medicament that is prepared from by the following weight proportion raw material:
9～15 parts of Fructus Psoraleae 6-10 part, Radix Puerariaes.
2, the pharmaceutical composition of treatment osteoporosis according to claim 1 is characterized in that: it is the medicament that is prepared from by the following weight proportion raw material:
10 parts of Fructus Psoraleaes, 10 parts of Radix Puerariaes.
3, the pharmaceutical composition of treatment osteoporosis according to claim 1 and 2, it is characterized in that: it is by Fructus Psoraleae total coumarins, Fructus Psoraleae total flavones, Radix Puerariae total flavones is an active component, adds the medicament that acceptable accessories or complementary composition are prepared from.
4, the pharmaceutical composition of treatment osteoporosis according to claim 3, it is characterized in that: described Fructus Psoraleae total coumarins is in psoralen, percentage by weight is not less than 24%, Fructus Psoraleae total flavones is in Coryfolin, and percentage by weight is not less than 26%; Radix Puerariae total flavones is in puerarin, and percentage by weight is not less than 70%.
5, according to the pharmaceutical composition of claim 3 or 4 described treatment osteoporosis, it is characterized in that: it is the medicament that is prepared from by the following weight proportion raw material:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones.
6, the pharmaceutical composition of treatment osteoporosis according to claim 5 is characterized in that: it is to be prepared from medicament by the following weight proportion raw material:
1 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1 part of Radix Puerariae total flavones.
7, according to the pharmaceutical composition of each described treatment osteoporosis of claim 1-6, it is characterized in that: described medicament is: tablet, capsule, pill, granule, slow releasing preparation.
8, the pharmaceutical composition of treatment osteoporosis according to claim 7 is characterized in that: described slow releasing preparation is prepared from by following weight proportion raw material and adjuvant:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones, slow releasing preparation adjuvant: 1～10 part.
9, the pharmaceutical composition of treatment osteoporosis according to claim 8 is characterized in that: described slow releasing preparation adjuvant is: a kind of or two or more mixing arbitrarily among ethyl cellulose EC, hydroxypropyl methylcellulose HPMC or the Sulisi surelease.
10, the pharmaceutical composition of treatment osteoporosis according to claim 9 is characterized in that: it is to be prepared from by following weight proportion raw material and adjuvant:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones, 0.5～10 part of EC, 0.2～1.0 part of HPMC.
11, the pharmaceutical composition of treatment osteoporosis according to claim 10 is characterized in that: it is to be prepared from by following weight proportion raw material and adjuvant:
1 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1 part of Radix Puerariae total flavones, 0.5 part of EC, HPMC0.5 part.
12, a kind of method for preparing the pharmaceutical composition of each described treatment osteoporosis of claim 1-11 comprises the steps:
A, get Fructus Psoraleae, add organic solvent extraction, Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture;
B, get Radix Puerariae, with 50～90% alcohol reflux, ethanol extract decompression recycling ethanol, filter, by adsorption resin column, with 30%～90% ethanol elution, collect ethanol elution, reclaim ethanol, drying promptly gets Radix Puerariae total flavones;
C, with the Fructus Psoraleae total coumarins of a, b step preparation and Fructus Psoraleae total flavones mixture, Radix Puerariae total flavones by following weight proportion, add acceptable accessories or complementary composition and be prepared into dosage form pharmaceutically commonly used:
1～5 part in Fructus Psoraleae total coumarins and Fructus Psoraleae total flavones mixture, 1～5 part of Radix Puerariae total flavones.
13, the preparation of drug combination method of treatment osteoporosis according to claim 12 is characterized in that: the described reflux, extract, concentration of alcohol of b step is 30% ethanol; The concentration of ethanol elution is 70% ethanol.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104147100A (en) * | 2014-08-19 | 2014-11-19 | 安徽九方制药有限公司 | Pueraria flavonid medical application |
| CN104800532A (en) * | 2015-04-13 | 2015-07-29 | 青岛华仁技术孵化器有限公司 | Calcium supplementing substances used in health food |
| CN105412848A (en) * | 2016-01-20 | 2016-03-23 | 青岛云天生物技术有限公司 | Medicine composition for treating osteoporosis and preparation method thereof |
| CN108464997A (en) * | 2018-05-03 | 2018-08-31 | 佛山科学技术学院 | A kind of compound Chinese patent medicine and preparation method thereof prevented, control osteoporosis |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1208072C (en) * | 2002-11-19 | 2005-06-29 | 贵州神奇制药有限公司 | 'Jinwugutong' capsule and its preparation technology |
-
2005
- 2005-05-08 CN CNB2005100208674A patent/CN100358543C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104147100A (en) * | 2014-08-19 | 2014-11-19 | 安徽九方制药有限公司 | Pueraria flavonid medical application |
| CN104800532A (en) * | 2015-04-13 | 2015-07-29 | 青岛华仁技术孵化器有限公司 | Calcium supplementing substances used in health food |
| CN105412848A (en) * | 2016-01-20 | 2016-03-23 | 青岛云天生物技术有限公司 | Medicine composition for treating osteoporosis and preparation method thereof |
| CN108464997A (en) * | 2018-05-03 | 2018-08-31 | 佛山科学技术学院 | A kind of compound Chinese patent medicine and preparation method thereof prevented, control osteoporosis |
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