CN102294076A - Biodegradable traditional Chinese medicine slow release implant for revascularization of ischemic lower limbs and preparation thereof - Google Patents
Biodegradable traditional Chinese medicine slow release implant for revascularization of ischemic lower limbs and preparation thereof Download PDFInfo
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Abstract
The invention relates to a biodegradable traditional Chinese medicine slow release implant for the revascularization of ischemic lower limbs and preparation thereof. The slow release implant comprises a hollow tube, wherein the tube is a single-layer one or a double-layer one and is prepared from biodegradable polymer loaded with traditional Chinese medicine for promoting revascularization; the biodegradable polymer is polycaprolactone, Pullulan polysaccharide, polylactic acid, polyglycolic acid, polylactic acid-polyglycolic acid copolymer, polycaprolactone-polylactic acid copolymer, polydioxane, hydroxypropyl methylcellulose, collagen, alginic acid or chitosan and derivates thereof; the mass ratio of the traditional Chinese medicine to the biodegradable polymer is 0.1-500mg:5-1000mg; and the traditional Chinese medicine is puerarin, salidroside, icariin, paeoniflorin, Danshensu, tanshinone, water extract of yellow sandalwood root or water extract of Mongolian snakegourd fruit. The slow release implant can promote the revascularization of ischemic lower limbs and improve the blood perfusion of the lower limbs. The preparation method provided by the invention is simple and easy to operate.
Description
Technical field
The present invention relates to a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing and preparation.
Background technology
Ischemic disease of lower extremity is one of disease of serious harm human health, and how it is caused that by stricture of artery due to the atherosclerosis or obturation it reaches modal symptom the earliest is intermittent claudication, and the ischemic rest pain can appear in severe patient, toes lack limb.Along with the raising day by day of living standard, the sickness rate of ischemic disease of lower extremity also is trend of rising year by year.In developing country, more than 50 years old among the crowd prevalence of ischemic disease of lower extremity up to 5%, the serious harm human health, country's huge medical care expenses of annual cost is used for this class disease of medical care, is threatening rapid development of national economy.
By cell growth factor, as blood vessel endothelial cell growth factor VEGF, basic fibroblast growth factor bFGF, transforming growth factors,type beta TGF-β etc., can stimulate revascularization, improve the tissue blood perfusion.But the half-life of somatomedin in blood circulation is very short, it only is tens of minutes, and the targeting poor location, being difficult to bring into play short angiogenesis function permanent, that continue, general is widely applied and can be caused adverse side effects such as blood pressure drops and potential short tumor mutation again.In addition, somatomedin costs an arm and a leg, and has limited its widespread usage in ischemic disease of lower extremity.
The development of Chinese medicine has a long history, and just records 200 and 300 various plants respectively in " Book of Songs " before 3,000 years and " elegant ", and wherein having much is medicinal plants.By succession accumulation in several thousand, people had certain understanding and understanding to function, curative effect and the side effect of Chinese crude drug, and still, the pharmacological action of Chinese medicine does not also obtain fully comprehensively excavating.For example: puerarin (Puerarin, C
21H
20O
9) be a kind of isoflavonoid that from legume pueraria lobata or Radix Puerariae rattan root, extracts, its main component is 4,7-dihydroxy-8-β-D glucose isoflavone.Since approval listing in 1992, people are more active always to the research of puerarin, in the pharmacopeia it is classified as a kind of vasodilator, clinical practice is in coronary heart disease, angina pectoris, myocardial infarction, arrhythmia, hypertension, arteria retina vein obstruction, sudden deafness, hyperlipidemia and hyperglycemia etc., and is evident in efficacy.Puerarin is generally the intravenous drip administration clinically, the administering mode of puerarin is intravenously administrable, filling stomach or oral administration etc. in the document zoopery, through sanguimotor effect, have only limited drug dose to arrive target site, and intravenously administrable easily cause untoward reaction such as acute intravascular hemolysis.
The biodegradability polymer support can be kept the support performance of support within a certain period of time, guarantees that tract is unobstructed, is degraded to innocuous substance at last and gets rid of external.After they finish its supporting role, can be organized absorption fully, side effect is less.Its mechanical property and degradation time can be controlled by the degree of polymerization and preparation process simultaneously, and preparation technology is simple, can pass through preparations such as extrusion by melting, solvent evaporation method, method of electrostatic spinning, so they has bigger potentiality in clinical practice.CN200710057459.5 discloses a kind of Biodegradable slow released medicated rack and preparation method that is used for myocardial blood transport reconstruction, the Biodegradable slow released medicated rack that is used for myocardial blood transport reconstruction, promptly with biodegradable polymers make hollow in the pipe, the outer surface of pipe is provided with medicine carrying biodegradable polymers thin layer in hollow, in hollow, run through on pipe and the medicine carrying biodegradable polymers thin layer and be provided with the hole that diameter is 0.2mm-1mm, medicine is that cell growth factor is or/and heparin, the Biodegradable slow released medicated rack that is used for myocardial blood transport reconstruction, can slowly discharge somatomedin or/and heparin, can stop cardiac muscle closed voluntarily and prevent thrombosis and promote hole wall to generate blood vessel endothelium that wherein biological degradation polyalcohol is polycaprolactone or polylactic acid or polyglycolic acid or polylactic acid-polyglycolic acid copolymer or polycaprolactone-copolymer of poly lactic acid or poly-dioxane.CN200910068305.5 also discloses a kind of locular wall myocardial blood transport reconstruction biodegradability medicine slow release stent.CN 200810053747.8 discloses a kind of Biodegradable slow released medicated rack and preparation method that is used for myocardial blood transport reconstruction, and timbering material is magnesium or magnesium alloy.
Do not have still at present that puerarin or other Chinese medicine are used for the treatment of the report, particularly puerarin of ischemic disease of lower extremity or medicine that other Chinese medicine is inserted in the thing as myocardial revascularization biodegradability medicament slow release does not also have this type of report.
Summary of the invention
The object of the present invention is to provide a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release to insert thing and preparation method.Lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release is inserted the generation that thing can promote ischemia lower limb new vessels, improves the lower extremity blood flow perfusion.Preparation method of the present invention is simple, easy operating.
A kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release provided by the invention is inserted thing and is comprised hollow pipe, the internal diameter of pipe is 1mm-6mm, wall thickness is 0.5mm-4mm, length is 0.5mm-200mm, the material of described pipe is the biodegradable polymers that load has the Chinese medicine that promotes angiogenesis, and described pipe is monolayer or bimetallic tube.
The mass ratio of described Chinese medicine and biodegradable polymers is 0.1-500mg: 5-1000mg.
Described polymer is polycaprolactone, pulullan polysaccharide, polylactic acid, polyglycolic acid, polylactic acid-polyglycolic acid copolymer, polycaprolactone-copolymer of poly lactic acid, poly-dioxane, hydroxypropyl emthylcellulose, collagen protein, alginic acid or chitosan and derivant thereof.
Described Chinese medicine is puerarin, rhodioside, icariin, peoniflorin, danshensu, TANSHINONES, yellow wingceltis root water extract or Fructus Trichosanthis fruit water extract.
The step that the preparation method that a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release provided by the invention is inserted thing comprises is as follows:
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, Chinese medicine is dispersed in the described biodegradable polymers solution, transfer in the mould, form thin film in the back fully in solvent evaporates, be cut into bar shaped, be rolled into pipe again, be cut into certain-length, promptly make a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing.Organic solvent is dichloromethane, chloroform, oxolane or acetone.Or
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, Chinese medicine is dispersed in the described biodegradable polymers solution, join in the syringe, make nanofiber by electrospinning device, adopt the tubular metal receptor to reel and be gathered into pipe, be cut into certain-length, promptly make a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing.Organic solvent is dichloromethane, chloroform, oxolane or acetone.Or
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, transfer in the mould, form thin film in the back fully in solvent evaporates, be cut into bar shaped, Chinese medicine is dispersed between two thin film, carrying out solvent by organic solvent fixes, be rolled into pipe again, be cut into certain-length, promptly make a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing.Organic solvent is dichloromethane, chloroform, oxolane or acetone.
Lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release of the present invention is inserted the generation that thing can promote ischemia lower limb new vessels, improves the lower extremity blood flow perfusion.Preparation method of the present invention is simple, easy operating.
Description of drawings
Fig. 1 inserts thing for a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release of the present invention and prepares sketch map.
Fig. 2 inserts the outer drug release behavior of object for a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release of the present invention.
Fig. 3 inserts thing for a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release of the present invention and promotes lower limb vascular regeneration effect (adductor longus m. is organized HE dyeing picture after inserting the thing effect).
Fig. 4 inserts thing for a kind of lower limb ischemia myocardial revascularization of the present invention biodegradability Chinese medicine slow release and promotes lower extremity blood flow perfusion effect (insert laser-Doppler blood flow imaging figure after the thing effect, the picture upside is inserted the group lower limb for inserting thing, and downside is normal lower limb).
The specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment.Reagent of the present invention is commercially available.
Embodiment 1
The preparation method that a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release is inserted thing comprises the steps:
Ratio in 0.5g: 20ml, with polylactic acid-polyglycolic acid copolymer (mol ratio 75/25, Mn10000, η 0.43, Chengdu Organical Chemical Co., Ltd., Chinese Academy of Sciences) be dissolved in the dichloromethane (Tianjin sky over the river chemical industry company limited), make biodegradable polymers solution, transfer in the mould of tetrafluoroethene, form thin film in the back fully, be cut into bar shaped in solvent evaporates, with 10mg puerarin (Puerarin, C
21H
20O
999.5%, Shanghai Mai Ruier chemical technology company limited) is dispersed between two thin film, by dichloromethane carry out solvent fixing after, being rolled into internal diameter is 2mm, wall thickness is the pipe of 2mm, is cut into length 10mm, promptly makes a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and inserts thing.
Equally, puerarin rhodioside (Salidroside, C
14H
20O
7), icariin (Icariin, C
33H
40O
15), peoniflorin (Paeoniflorin, C
23H
28O
11), danshensu (Danshensu Salvianic acid A, C
9H
10O
5), TANSHINONES (C
18H
12O
3), yellow wingceltis (Dalbergia hupeana) root water extract or Fructus Trichosanthis (Trichosanthes cucumerina L.) fruit water extract substitute, and can form new embodiment, obtain degradability Chinese medicine slow release and insert thing.
The polylactic acid-polyglycolic acid copolymer substitutes with polycaprolactone, pulullan polysaccharide, polylactic acid, polyglycolic acid, polycaprolactone-copolymer of poly lactic acid, poly-dioxane, hydroxypropyl emthylcellulose, collagen protein, alginic acid or chitosan and derivant thereof, can form new embodiment, obtain degradability Chinese medicine slow release and insert thing.
Methylene chloride substitutes with chloroform, oxolane or acetone, can form new embodiment, obtains degradability Chinese medicine slow release and inserts thing.
Embodiment 2
Insert the extracorporeal releasing experiment of thing:
Content of puerarin adopts high performance liquid chromatography to measure.Chromatographic column: the C18 post (200mm * 4.6mm, 5 μ m, Kromasil); Mobile phase: methanol-water (25: 75); Flow velocity is 1ml/min, detects wavelength 250nm, 30 ℃ of column temperatures, sample size 20 μ L.Under above-mentioned high-efficient liquid phase chromatogram condition, be not less than 3000 by the puerarin peak theory of computation number of plates.It is an amount of that precision takes by weighing the puerarin reference substance, adds the mobile phase dissolving.And make the solution of 1.57,3.14,7.85,15.7,62.8,94.2,125.6,157 μ g/ml, and under above-mentioned chromatographic condition, measure respectively, write down its peak area.With peak area concentration is carried out regression analysis, get regression equation.
To insert thing and be put in the sample cell that contains 5ml PBS buffer (pH7.4), and place 37 ℃ gas bath constant temperature oscillator, rotating speed is 100r/min.Timing sampling, and additional PBS buffer with volume.Calculate content of puerarin by equation of linear regression, and try to achieve the cumulative release percentage rate.Each sample retest three times.
Embodiment 3
Zoopery
(1) foundation of rat acute lower limb ischemia model
SPF level wistar rat, male, body weight 220-250g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., and credit number is SCXK (capital) 2009-0004.The raising condition: room temperature 23-25 degree, put in the rat Rotating Stainless Steel Cage tool and raise, freely take the photograph water and ingest.12h fasting before the rat art is freely drunk water.10% chloral hydrate solution 0.3ml/100g intraperitoneal injection of anesthesia, it is fixing to lie on the back, the conventional preserved skin sterilization of left lower extremity, the skin of longitudinal incision left lower extremity center, from groin to knee joint.Isolate femoral artery under groin, until knee joint, the ligation femoral artery is in all branches of thigh portion from inguinal ligament for scope, and femoral artery is cut off under inguinal ligament.Art finishes, and sews up the incision.Determine that by the laser-Doppler blood flow imaging lower limb ischemia model prepares successfully.
(2) muscle punches and inserts inserting of thing
Modeling success rat is divided at random that blank is inserted the thing group and puerarin is inserted the thing group, 6/every group.Rat lower limb ischemia adductor longus m. adopts high-speed drilling device (13000r/min) punching, inserts one piece in each duct and inserts thing (embodiment 3), sew up wound immediately.
(3) the laser-Doppler blood flow imaging is analyzed
Respectively at postoperative 1 day, 2 weeks, 4 weeks, 6 weeks, with rat 10% chloral hydrate solution 0.3ml/100g intraperitoneal injection of anesthesia, after 37 ℃ of heated at constant temperature make its pair lower limb temperature constant relatively.Animal is got supine body position, row laser-Doppler blood flow detection.
(4) histologic analysis
Put to death animal in 6 weeks of postoperative, take off limb ischemic region tissue, 40g/L formaldehyde fixed 24h, dehydration, waxdip, embedding, the 5 μ m that cut into slices, conventional row hematoxylin-eosin (HE) dyeing, detailed process is: 1. brazilwood extract dyeing 10min, washing from the beginning; 2. 1% hydrochloride alcohol color separation, color separation limit, limit microscopy is reddish violet to examining, and tap water is washed till nucleus and becomes blue, and distilled water is cleaned alkaline matter; 3. 0.5-1% Yihong aqueous solution is washed 10-20 second, washing from the beginning, distillation washing; 4. 95% ethanol color separation is rinsed and is washed 15 seconds; 5. 95%, 100% ethanol gradient dehydration 10min, dimethylbenzene dehydration 1-2min; 6. resinene mounting, ultramicroscopic observation.
Result of the test
(1) vitro drug release behavior
The puerarin standard substance are done linear regression with peak area to sample size, and regression equation is: y=0.0569x+0.0458 (r=0.9999), puerarin sample size and peak area in 0.0314-3.14 μ g scope has good linear relationship as can be known.Inserting in the thing research of puerarin release in vitro carries out in 37 ℃ of gas bath constant temperature oscillators.Fig. 2 is the release profiles of medicine, and as can be seen from the figure, puerarin is slowly stable to be continued to discharge, and along with the degraded of inserting the beyond the region of objective existence layer material, the puerarin that discharges after 25 days is near 70%.Illustrate that this medicine inserts the purpose that thing can reach medicament slow release, the drug release longer duration is degraded fully until outer drug-loaded layer.
(2) nascent blood vessel density quantitative analysis
Insert thing and inserted for 2,4,6 weeks, check the morphological assessment nascent blood vessel density by histochemistry.At each time point, puerarin is inserted thing and is inserted the vessel density of group and all insert thing apparently higher than blank and insert group.Insert thing in blank and insert group, vessel density reduces gradually along with the prolongation of time.Insert thing for puerarin and insert group, when inserting for 4 weeks, vessel density reaches summit (189.56 ± 8.23/mm
2); When inserting for six weeks, vessel density reduces (102.45 ± 10.46/mm
2), but vessel lumen chap tube wall thickening (Fig. 3).
(3) the laser-Doppler lower extremity blood flow detects
12 rats are successfully caused the left lower extremity ischemia model, perform the operation back first day, and blood flow all is obvious reduction trend.Subsequently, insert thing at puerarin and insert group, blood flow recovers gradually, and is in rising trend, and quantitative analysis results shows the 6th week of postoperative, and blood perfusion reaches 85.46 ± 3.59%, and blood perfusion obviously improves.Insert group and insert thing, the basic no change of ischemia left lower extremity blood perfusion amount, blood perfusion do not improve (Fig. 4) in blank.
Claims (9)
1. a lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release is inserted thing, it is characterized in that it comprises hollow pipe, the internal diameter of pipe is 1mm-6mm, wall thickness is 0.5mm-4mm, length is 0.5mm-200mm, the material of described pipe is the biodegradable polymers that load has the Chinese medicine that promotes angiogenesis, and described pipe is mono-layer tube or bimetallic tube.
2. the thing of inserting according to claim 1, the mass ratio that it is characterized in that described Chinese medicine and biodegradable polymers is 0.1-500mg: 5-1000mg.
3. the thing of inserting according to claim 1, the mass ratio that it is characterized in that described Chinese medicine and biodegradable polymers is 0.1-50mg: 5-100mg.
4. the thing of inserting according to claim 1, the internal diameter that it is characterized in that described pipe is 2mm, wall thickness is the pipe of 2mm, length 10mm.
5. the thing of inserting according to claim 1 is characterized in that described biodegradable polymers is polycaprolactone, pulullan polysaccharide, polylactic acid, polyglycolic acid, polylactic acid-polyglycolic acid copolymer, polycaprolactone-copolymer of poly lactic acid, poly-dioxane, hydroxypropyl emthylcellulose, collagen protein, alginic acid or chitosan and derivant thereof.
6. the thing of inserting according to claim 1 is characterized in that described Chinese medicine is puerarin, rhodioside, icariin, peoniflorin, danshensu, TANSHINONES, yellow wingceltis root water extract or Fructus Trichosanthis fruit water extract.
7. the thing of inserting according to claim 1 is characterized in that described Chinese medicine is puerarin.
8. the described lower limb ischemia myocardial revascularization of claim 1 a biodegradability Chinese medicine slow release is inserted the preparation method of thing, it is characterized in that the step that comprises is as follows:
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, Chinese medicine is dispersed in the described biodegradable polymers solution, transfer in the mould, form thin film in the back fully in solvent evaporates, be cut into bar shaped, be rolled into pipe again, be cut into certain-length, promptly make a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing; Or
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, Chinese medicine is dispersed in the described biodegradable polymers solution, join in the syringe, make nanofiber by electrospinning device, adopt the tubular metal receptor to reel and be gathered into pipe, be cut into certain-length, promptly make a kind of lower limb ischemia myocardial revascularization biodegradability Chinese medicine slow release and insert thing; Or
In proportion biodegradable polymers is dissolved in the organic solvent, make biodegradable polymers solution, transfer in the mould, form thin film in the back fully in solvent evaporates, be cut into bar shaped, Chinese medicine is dispersed between two thin film, carrying out solvent by organic solvent fixes, be rolled into pipe again, be cut into certain-length, get final product.
9. preparation method according to claim 8 is characterized in that described organic solvent is dichloromethane, chloroform, oxolane or acetone.
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| CN102697721A (en) * | 2012-07-04 | 2012-10-03 | 西南民族大学 | Salidroside segmented copolymer lipid nanoparticle preparation |
| CN105288831A (en) * | 2014-06-03 | 2016-02-03 | 辽宁省计划生育科学研究院 | Hollow tubular biodegradable medication system and preparation method thereof |
| CN105687216A (en) * | 2016-01-21 | 2016-06-22 | 重庆大学 | Medicine for treating lower limb ischemic disease |
| CN107158393A (en) * | 2017-06-20 | 2017-09-15 | 苏州乔纳森新材料科技有限公司 | A kind of copolymerized material of pulullan polysaccharide PLA and preparation method thereof |
| CN108096630A (en) * | 2018-01-29 | 2018-06-01 | 暨南大学 | A kind of polylactic acid base tissue scaffold design and preparation method and application for carrying icariin and Deferoxamine |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697721A (en) * | 2012-07-04 | 2012-10-03 | 西南民族大学 | Salidroside segmented copolymer lipid nanoparticle preparation |
| CN102697721B (en) * | 2012-07-04 | 2014-02-19 | 西南民族大学 | Salidroside segmented copolymer lipid nanoparticle preparation |
| CN105288831A (en) * | 2014-06-03 | 2016-02-03 | 辽宁省计划生育科学研究院 | Hollow tubular biodegradable medication system and preparation method thereof |
| CN105288831B (en) * | 2014-06-03 | 2019-11-08 | 辽宁省计划生育科学研究院 | A kind of biodegradation type drug delivery system of hollow tubular and preparation method thereof |
| CN105687216A (en) * | 2016-01-21 | 2016-06-22 | 重庆大学 | Medicine for treating lower limb ischemic disease |
| CN105687216B (en) * | 2016-01-21 | 2018-08-03 | 重庆大学 | Treat the drug of ischemic disease of lower extremity |
| CN107158393A (en) * | 2017-06-20 | 2017-09-15 | 苏州乔纳森新材料科技有限公司 | A kind of copolymerized material of pulullan polysaccharide PLA and preparation method thereof |
| CN108096630A (en) * | 2018-01-29 | 2018-06-01 | 暨南大学 | A kind of polylactic acid base tissue scaffold design and preparation method and application for carrying icariin and Deferoxamine |
| CN108096630B (en) * | 2018-01-29 | 2020-09-04 | 暨南大学 | Icariin and deferoxamine-loaded polylactic acid-based bone tissue scaffold and preparation method and application thereof |
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Application publication date: 20111228 |