CN104288219A - Preparation method and clinical application of fructus psoraleae derivative transdermal absorbent - Google Patents
Preparation method and clinical application of fructus psoraleae derivative transdermal absorbent Download PDFInfo
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- CN104288219A CN104288219A CN201410573329.7A CN201410573329A CN104288219A CN 104288219 A CN104288219 A CN 104288219A CN 201410573329 A CN201410573329 A CN 201410573329A CN 104288219 A CN104288219 A CN 104288219A
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- C07—ORGANIC CHEMISTRY
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- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
- A61K31/37—Coumarins, e.g. psoralen
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/487—Psoralea
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A—HUMAN NECESSITIES
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/38—Cellulose; Derivatives thereof
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
The invention relates to a preparation method and clinical application of a fructus psoraleae derivative transdermal absorbent. The preparation method comprises the following steps: extracting fructus psoraleae powder by using ethanol, peracetylating the extract to prepare a derivative; adding 2-10g of carboxymethylcellulose (or polyvinyl alcohol), 1g of glycerin, 1g of castor oil and 1-2g of azone into the peracetylated derivative of the fructus psoraleae extract; weighing totally 10g of sodium alga acid, adding 100ml of water and 2-10g of carboxymethyl cellulose so as to prepare the transdermal absorbent. The preparation is simple in preparation method, easily soluble in water, wide in raw material source and low in cost and does not have any toxic effect.
Description
Technical field
The invention belongs to clinical medicine medication, relate to a kind of medicine being applicable to fracture, bone does not connect, femur head necrosis, osteoporotic treatment and prevention specifically.
Background technology
Union of fracture is an extremely complicated biological process, and by the impact of factors, how early promotion union of fracture is the emphasis problem of orthopedics department area research always.In recent years, greater advance is obtained to delayed union of fracture, disunion Therapy study.Treatment by Chinese herbs is fractured, with a long history, and experience is unique, is widely used.From traditional Orally taken and externally, develop into the injection extracting active ingredient and make from single or compound recipe, determined curative effect easy with it and show its superiority.But the therapeutic mechanism of Chinese medicine promotion union of fracture is a very complicated process.Along with going deep into of research, the impact of Chinese medicine on it receives the concern of more and more scholar.
Fructus Psoraleae is the dry fruit of legumes psoraleae, containing plurality of active ingredients.In vitro tests shows, the increase of the absorption lacuna that Fructus Psoraleae energy control separation osteoclast is formed on osteocomma and expansion, illustrates that it has inhibitory action to separation osteoclast.Research finds, psoralen has obvious facilitation to proliferation of rat osteoblasts and alkaline phosphatase activities.
The derivant of psoralen comprises 5-MOP (5-MOP), 8-methoxypsoralen (8-MOP) and some substituents, existing by wherein several typical structure row are as follows: Pabulenol; Saxalin; Wang Yincao; Aqua oxidation peucedanin; Angelol; Some hydroxy derivatives of formic acid oxidation peucedanin.
Woods is lifted to select etc. and measures Fructus Psoraleae injection to the impact of proliferation of rat osteoblasts with mtt assay, result display Primary osteoblast cells forms fibrocyte colony gradually, have fibroblast cell type to show, go down to posterity osteoblast gradually embed by the substrate of emiocytosis, formed calcification.Fructus Psoraleae has remarkable facilitation to the osteoblastic propagation of neonate rat, shows to utilize trypsin digestion separation and Culture can go out that purity is high, quantity is more and the osteoblast that incubation time obviously shortens.The effect of Fructus Psoraleae osteoporosis is with the osteoblastic quantity of its increase and promote that osteoblastic multiplication capacity is relevant.[Lin Juze, Chen Shengkai, Luo Jiadong, the impact [J] that Fructus Psoraleae injection is bred rat osteoblast in vitro. the traditional Chinese medical science bonesets, and 2004,16 (6): 6.]
Summary of the invention
The object of the present invention is to provide a kind of preparation method of full acetylated Psoralen derivatives preparation capable of permeating skin.Be to provide a kind of preparation method simple, soluble in water, raw material sources are treated widely and are prevented the medicine of fracture.
Advantage of the present invention is that preparation method is simple, and raw material sources are wide, with low cost, have no side effect.
The preparation method of Psoralen derivatives preparation capable of permeating skin of the present invention, comprises the following steps:
Extracting method: get Malaytea Scurfpea Fruit (crossing 40 mesh sieves) 100g, add 10 times amount 70% ethanol, soak 20hr, heating and refluxing extraction 2h, extract 2 times, each 40min.Merge 2-3 filtrate, decompression and solvent recovery, concentrated, dry, dry thing dissolve with methanol, activated carbon removing impurities matter removes methanol solvate again, concentrated, dry, obtains psoralen extract.
Prepared by Psoralen derivatives: by psoralen extract 30g, acetic anhydride 200ml and sulfamic acid 2g, react 30min under being mixed in room temperature, then continues temperature to rise to 60-90 DEG C of reaction 20min and complete reaction.To the end that reacts completely, cross and filter unreacted acetic anhydride.Add ethyl acetate 200ml in product to rinse, sodium bicarbonate 5% (10mL × 2) rinses.Then dry, filter, concentrating under reduced pressure obtains product.
The preparation of the full acetylated derivant preparation capable of permeating skin of Fructus Psoraleae:
1. scheme 1: the prescription of Fructus Psoraleae full acetylated derivant preparation capable of permeating skin: the full acetylated derivant 10g of Fructus Psoraleae, carboxymethyl cellulose 2-10g, glycerol 1g, Oleum Ricini 1g, azone 1-2g.Take Psoralen derivatives 10g, add water 100ml, carboxymethyl cellulose 2-10g, be stirred to clear and bright glue, under stirring, add azone 1-2g, glycerol 1g, Oleum Ricini 1g successively, mixing, regulate pH value to be about 7-7.5 with sodium bicarbonate, ultrasonic 15min.Coat during use on gauze, cover therapentic part.
2. scheme: 2: the prescription of Fructus Psoraleae full acetylated derivant preparation capable of permeating skin: the full acetylated derivant 10g of Fructus Psoraleae, polyvinyl alcohol 2-10g, glycerol 1g, Oleum Ricini 1g, azone 1-2g.Take Psoralen derivatives 10g, add water 100ml, polyvinyl alcohol 2-10g, be stirred to clear and bright glue.Add azone 1-2g, glycerol 1g, Oleum Ricini 1g under stirring successively, mixing, regulate pH value to be about 7-7.5 with sodium bicarbonate, ultrasonic 15min.Coat during use on gauze, cover therapentic part.
Described pharmaceutical composition, promotes the application in the treatment of human or animal's union of fracture in preparation.
In order to further verify the curative effect of finished product preparation of the present invention, the medicine prepared in above-mentioned concrete preparation method 1-3 has been carried out corresponding animal experiment by us, existing that report the test is as follows.
Reagent: bone morphogenesis protein-2 (BMP-2) antibody is purchased from Beijing Bo Aosen Bioisystech Co., Ltd
Prepared by animal grouping model: 40 no-special pathogen kunming mices (by Medical School of Zhengzhou University Experimental Animal Center [Scxk (Henan) 2010-0002]) provide.The scope of Mouse Weight is 25 ~ 30g, 3 monthly ages, male and female half and half.
40 mices are divided into treatment group, model group two groups at random, often organize 20.Treatment group, model group mice are with 20% urethanes 5mg/kg intraperitoneal injection of anesthesia, and exposed Rats femur, cuts femur with hacksaw, ooze out for degree, sewed up by wound to reach bone marrow.Then treatment group mice limbs external application preparation capable of permeating skin near wound, qd, successive administration 7-14d and continuously 3d During Intramuscular Injection of Penicillin with infection.Mice does not deal with model group.
Collection of specimens and histology process
40 mices are put to death at the 1st, 2 weekends respectively, get femur and do histology's preparation.It is 7.2 that femur 4%PFA adds 0.1M phosphate buffer adjustment pH value, and room temperature fixes 3d.Then adding 0.1M phosphate buffer adjustment pH value with 20%EDTA is 7.2, soaks 8-10d decalcification, then through series of ethanol dehydration, paraffin embedding, is cut into 5 μm of sheets along the capable thickness of femoral shaft long axis direction, and partial row HE dyes.Get 5 femur sheets at random, observe under × 400 Olympus FV1000 amplifying laser scanning confocal microscopes respectively.
Immunohistochemical staining
After sacrifice centered by callus, comprise each 1cm of upper and lower epiphysis and take out specimen, insert decalcification 10-15d in 5% ethylenediaminetetraacetic acid (EDTA) liquid, ethanol dewaters step by step, transparent, waxdip, paraffin embedding, 5 μm of thick serial section, ABC method immunohistochemical staining (ABC test kit is provided by Vector company).Normal group bone group BMP-McAb dyes as negative, and after fracture, callus tissue staining is positive.
Statistical procedures adopts SPSS10.0 software to carry out statistical analysis.Each group of data with
represent, compare employing independent samples t test between two groups, P < 0.05 represents that difference has statistical significance.
Result and conclusion
The immuno positive Product Expression of 2.1BMP-2 albumen in bone matrix and in osteoblast kytoplasm, in brown color.Adopt semiquantitative method to judge the expression of BMP-2 albumen: to select 10 high power fields, the positive reaction cell number < 25% of colour developing is+, 25 ~ 50% are ++, > 75% is +++, without obvious positive reaction cell be-.The statistical analysis of enumeration data adopts X 2 test.Touchstone α=0.05.
The expression of 2.2 postoperative 7d, two groups of BMP-2 albumen compares
Table 1 sodium alginate derivant aqueous solution is on the impact (2W, n=20) of BMP-2 content in fracture Mouse Bone
The blank group of P < 0.05, treatment group vs
Accompanying drawing explanation
Fig. 1 illustrates: the expression (sp of postoperative 7d BMP-2 albumen, 400 ×), Thallus Laminariae (Thallus Eckloniae) concocts rear product treatment group, the right femur middle-end of fracture mice, and the result of endochylema brown color engrain appears in immunohistochemical staining, (-) 0 is routine, (+) 2 is routine, (++) example, (+++) example, positive rate higher than model group, two groups of mice significant differences.
Postoperative 7d, treatment group BMP-2 protein expression is higher than blank group (p < 0.05).Result has statistical significance.
Fig. 2 illustrates: postoperative 14d, treatment group BMP-2 protein expression is higher than blank group (p < 0.01).Result has statistical significance.
Detailed description of the invention
Below the specific embodiment of content of the present invention, for set forth in present specification want the technical scheme of technical solution problem.
Object of the present invention realizes by following technique measures:
Embodiment 1
Extracting method: get Malaytea Scurfpea Fruit (crossing 40 mesh sieves) 100g, add 10 times amount 70% ethanol, soak 20hr, heating and refluxing extraction 2h, extract 2 times, each 40min.Merge 2-3 filtrate, decompression and solvent recovery, concentrated, dry, dry thing dissolve with methanol, activated carbon removing impurities matter removes methanol solvate again, concentrated, dry, obtains psoralen extract.
Fructus Psoraleae fat full acetylated derivant preparation: by Fructus Psoraleae extract 30g, acetic anhydride 200ml and sulfamic acid 2g, reacts 30min under being mixed in room temperature, then temperature is continued rise to 60-90 DEG C of reaction 20min and complete reaction.To the end that reacts completely, cross and filter unreacted acetic anhydride.Add ethyl acetate 200ml in product to rinse, sodium bicarbonate 5% (10mL × 2) rinses.Then dry, filter, concentrating under reduced pressure obtains the full acetylated derivant of product Fructus Psoraleae.
The preparation of the full acetylated derivant preparation capable of permeating skin of Fructus Psoraleae:
The prescription of Fructus Psoraleae full acetylated derivant preparation capable of permeating skin: the full acetylated derivant 10g of Fructus Psoraleae, carboxymethyl cellulose 2-10g, glycerol 1g, Oleum Ricini 1g, azone 1-2g.Take Psoralen derivatives 10g, add water 100ml, carboxymethyl cellulose 2-10g, be stirred to clear and bright glue.Add azone 1-2g, glycerol 1g, Oleum Ricini 1g under stirring successively, mixing, regulate pH value to be about 7-7.5 with sodium bicarbonate, ultrasonic 15min.Coat during use on gauze, cover therapentic part.
Embodiment 2
Extracting method: get Malaytea Scurfpea Fruit (crossing 40 mesh sieves) 100g, add 10 times amount 70% ethanol, soak 20hr, heating and refluxing extraction 2h, extract 2 times, each 40min.Merge 2-3 filtrate, decompression and solvent recovery, concentrated, dry, dry thing dissolve with methanol, activated carbon removing impurities matter removes methanol solvate again, concentrated, dry, obtains psoralen extract.
Fructus Psoraleae full acetylated derivant preparation: by Fructus Psoraleae extract 30g, acetic anhydride 200ml and sulfamic acid 2g, reacts 30min under being mixed in room temperature, then temperature is continued rise to 60-90 DEG C of reaction 20min and complete reaction.To the end that reacts completely, cross and filter unreacted acetic anhydride.Add ethyl acetate 200ml in product to rinse, sodium bicarbonate 5% (10mL × 2) rinses.Then dry, filter, concentrating under reduced pressure obtains the full acetylated derivant of product Fructus Psoraleae.
The prescription of Fructus Psoraleae full acetylated derivant preparation capable of permeating skin: the full acetylated derivant 10g of Fructus Psoraleae, polyvinyl alcohol 2-10g, glycerol 1g, Oleum Ricini 1g, azone 1-2g.Take Psoralen derivatives 10g, add water 100ml, polyvinyl alcohol 2-10g, be stirred to clear and bright glue.Add azone 1-2g, glycerol 1g, Oleum Ricini 1g under stirring successively, mixing, regulate pH value to be about 7-7.5 with sodium bicarbonate, ultrasonic 15min.Coat during use on gauze, cover therapentic part.
Claims (3)
1. the preparation method of psoralen derivative preparation capable of permeating skin, is characterized in that psoralen is through ethanol extraction, decompression and solvent recovery, concentrated, dry; Dry thing dissolve with methanol, activated carbon removing impurities matter removes methanol solvate again, concentrated, dry.By psoralen extract, acetic anhydride and the mixing of catalyst sulfamic acid are carried out full acetylatedly being obtained by reacting the full acetylated derivant of psoralen.
2. the prescription of psoralen full acetylated derivant preparation capable of permeating skin: the full acetylated derivant 10g of Fructus Psoraleae, carboxymethyl cellulose (or polyvinyl alcohol) 2-10g, glycerol 1g, Oleum Ricini 1g, azone 1-2g.
3. the full acetylated Psoralen derivatives preparation capable of permeating skin of right 2 promotes the application in the treatment of human or animal's union of fracture in preparation.
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101057885A (en) * | 2006-04-20 | 2007-10-24 | 周亚伟 | Medicinal composition for treating osteoporosis and its preparations and application |
CN103816110A (en) * | 2013-08-19 | 2014-05-28 | 郭宏昌 | Preparation method of drug transdermal absorbent promoting fracture healing |
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Patent Citations (2)
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CN101057885A (en) * | 2006-04-20 | 2007-10-24 | 周亚伟 | Medicinal composition for treating osteoporosis and its preparations and application |
CN103816110A (en) * | 2013-08-19 | 2014-05-28 | 郭宏昌 | Preparation method of drug transdermal absorbent promoting fracture healing |
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