CN101658484A - Preparation method and application of chitosan copolymer carrier micelle modified by cholesterol formyl chloride - Google Patents
Preparation method and application of chitosan copolymer carrier micelle modified by cholesterol formyl chloride Download PDFInfo
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Abstract
The invention relates to a preparation method and an application of chitosan copolymer carrier micelle modified by cholesterol formyl chloride, which can effectively solve the preparation problem of chitosan copolymer carrier micelle modified by cholesterol formyl chloride. The technical scheme comprises the steps of: 1. purifying chitosan; 2. preparing the chitosan copolymer modified by cholesterol formyl chloride; 3. separating and purifying the chitosan copolymer modified by cholesterol formyl chloride; and 4 preparing the chitosan copolymer carrier micelle modified by cholesterol formyl chloride. The method has the characteristic of preparing a colloid solution with stable thermodynamics by self-assembling in a water medium, has simple preparation process and good effect and is an innovation in the medical field.
Description
One, technical field
The present invention relates to medical domain, particularly a kind of preparation method of chitosan copolymer carrier micelle of modified by cholesterol formyl chloride and application.
Two, background technology
Chitosan is de-acetyl chitin and a kind of biopolymer of obtaining; it is the main derivant of chitin; chitin and chitosan are nitrogenous polysaccharose substances; it also is the unique natural alkaline polysaccharide of free amine group that contains of occurring in nature; have different physiological roles such as antibacterial, anticancer, blood fat reducing, enhance immunity; but and have nontoxic, non-stimulated, non-immunogenicity, no heat source response, characteristics such as haemolysis, no mutagenicity and natural degradation, the favorable tissue compatibility, have great medical application and be worth.Over nearly 20 years, along with the discovery of the multiple derivatives active of chitosan, the whole world is very active to the research of this series products, and its application is also constantly widened.1991, the medical circle in the U.S., Europe, scientific and technological circle, nutraceutical research institution were with its 6th vital principle as needed by human after protein, sugar, fat, vitamin, mineral.1997, the research and development problem of chitosan was listed China State Scientific and Technological Commission State Key Task 95 project in, after be put into China country 863 Program, and obtained lot of research.
Yet the height-oriented and intermolecular intensive hydrogen bond action of chitosan interior molecules has limited its application.There are free amino and activity hydroxy in the chitosan molecule, therefore are easy to chitosan is carried out structure of modification, obtain the enhanced derivant of different structure, difference in functionality and dissolubility.The mode of derivatization mainly contains: acyl groupization, carboxylated, alkylation, esterification, hydroxylating and etherificate etc.But the many modification groups of chitosan make its derivant have many particular performances, so the concern of extremely domestic and international drug research person and pharmacy corporation.Be particularly useful for the embedding and the release of bioactive macromolecule medicines such as polypeptide, protein and nucleoside medicine.Aspect pharmaceutical preparation, as be applied to slow release, controlled release preparation, had suitable the research degree of depth and range.Chitosan nanoparticles being modified, made microgranule have targeting to some organ, tissue or cell, will further widen the using value of chitosan, is present research focus.
Tumor is the major disease that threatens human health always, and its Drug therapy key is to improve the tumor-selective of medicine, reduces its gathering at non-target site, thereby solves problems such as therapeutic effect is poor, toxic and side effects is huge.By the suitable carriers technology, be to solve cancer chemotherapy therapeutic effect difference and reduce one of important means of its toxic and side effects with the direct targeting pathological tissues of medicine (organ), cell.
Along with the development of biological nano technology, tumour medicine is contained in microcarrier, the performance nanometer size effect easily sees through various biological barriers, and can be loaded with more medicine, and this has become the focus of the novel drug-supplying system research of cancer therapy drug.Wherein, biodegradable amphipathic copolymer drug-loading system is extremely gazed at.Amphipathic being meant has hydrophobic group and hydrophilic group two parts simultaneously in the molecular structure.Copolymer micelle is that this amphipathic block or graft copolymer autohemagglutination in aqueous medium forms, and has nucleocapsid structure, the micelle diameter the nanoscale scope (＜200nm).Its hydrophobic chain segment constitutes micellar kernel, and hydrophilic segment forms micellar shell.Hydrophobicity can be the effect that insoluble drug provides bank, the hydrophilic adventitia keeps the stability of micelle in aqueous environments, and can carry out physicochemical property and modify reaching specific purpose, as micellar active targeting etc., form amphipathic copolymer nano micelle medicine carrying system.
This drug-loading system has following characteristic: (1) critical micelle concentration is low, the thermodynamic stability height; (2) but the amphipathic copolymer drug-loading system of shell-core structure self assembly in water or body fluid forming nano-micelle, micellar skin is the shell that hydrophilic section forms, internal layer is the nuclear that hydrophobic section forms, shell-core structure is the peculiar configuration of amphipathic copolymer; (3) the targeting tumor tissues is woven with abundanter blood capillary than normal group, and capillary wall has bigger hole, after cancer therapy drug is made amphipathic copolymer carrier micelle, because the carrier micelle size is little, specific surface area is big, its infiltration to tumor cell strengthens, and the drug level in the tumor cell will be apparently higher than blood drug level, and this has demonstrated the targeting of preparation; (4) after the copolymer micelle dehydration, redissolve, promptly recover former micelle structure in solvent.
In recent years, copolymer micelle has caused that in pharmaceutical preparation, biomedicine and field of polymer technology people pay attention to widely.Copolymer micelle has many advantages as a kind of drug delivery system: (1) copolymer micelle is because its copolymer dissolubility is little, so critical micelle concentration is very low, and its hydrophobic core core is more stable, so copolymer micelle can be difficult for depolymerizing through dilution.Therefore it has more protection and shielding action to medicine, can avoid the decomposition of medicine to a certain extent, keeps stability of drug, reduces the toxicity of medicine; (2) compare with liposome, the drug loading of copolymer micelle is higher; (3) multiformity of copolymer material makes with the copolymer to be the also variation of pharmaceutical preparation of carrier, can satisfy various user demands; (4) because the micelle particle diameter is very little, so can pass reticuloendothelium system, blood brain barrier, also can make the intestines and stomach mucosa etc. to its good absorption, the position that arrival large scale particle can't pass through reaches the purpose of passive target; (5) amphipathic copolymer is actually a kind of molecules surfactant, with the difference of low-molecular-weight surfactant be, the nuclear that low-molecular-weight surfactant forms is liquid, and the nuclear of copolymer micelle is solid-state, solid-state nuclear very helps the slow release of medicine, and the pH value by adjusting material, dissolubility, Zeta potential etc. can be controlled medicine release in vivo.But do not see the open report of method of the chitosan copolymer carrier micelle of relevant preparation modified by cholesterol formyl chloride so far.
Three, summary of the invention
At above-mentioned situation, for overcoming the prior art defective, the present invention's purpose just provides a kind of preparation method and application of chitosan copolymer carrier micelle of modified by cholesterol formyl chloride, can effectively solve the preparation problem of cholesterol beautify chitosan copolymer carrier micelle, the technical scheme of its solution is, 1, the purification chitosan; 2, the chitin copolymer of preparation modified by cholesterol formyl chloride; 3, the separation and purification of the chitin copolymer of modified by cholesterol formyl chloride; 4, the chitosan copolymer carrier micelle of preparation modified by cholesterol formyl chloride.The present invention has the characteristic that forms a kind of thermodynamically stable colloid solution in the aqueous medium self assembly, and preparation is simple, and is effective, is the innovation in the medical domain.
Four, the specific embodiment
Below in conjunction with concrete condition the specific embodiment of the present invention is elaborated.
The chitin copolymer of the modified by cholesterol formyl chloride among the present invention, its structural formula is as follows:
Wherein m, n are the degree of polymerization, m+n≤20, and the molecular weight of chitosan is 3～50kDa, and deacetylation is 70～100%, and the part amino on the chitosan chain is replaced by cholesterol formyl chloride;
The method of modified by cholesterol formyl chloride Preparation of Chitosan of the present invention, realized by following steps:
1, the purification of chitosan: the chitosan crude product of getting molecular weight 3～50kDa, glacial acetic acid solution (the solid-liquid mixing usually that adds 65～85 times the 0.1M (M is a molar concentration) of chitosan weight, solids is counted with weight g, liquids is counted with volume ml, volume weighs, therefore, the glacial acetic acid solution that chitosan adds 0.1M also can be written as " get the chitosan crude product of molecular weight 3～50kDa; add the glacial acetic acid solution of 65～85 times 0.1M of chitosan bulking value, as follows), stirring and dissolving; filter; add sodium hydroxide (NaOH) solution of 0.2M in the filtrate, in and glacial acetic acid, chitosan is precipitated fully; filter; with the deionized water wash filter cake to pH value 6～7, lyophilization obtains the chitosan of purification;
2, the preparation of the chitin copolymer of modified by cholesterol formyl chloride:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain, the chitin copolymer that the synthetic cholesterol formyl chloride is modified, step is: the chitosan and the cholesterol formyl chloride that take by weighing purification, chitosan is dispersed in volume ratio: 1～3: in 3～1 the dimethyl sulfoxide and chloroformic solution, the concentration of chitosan is 5～15mg/ml in the solution, add triethylamine 1ml～2ml and carry out catalysis and magnetic agitation, get mixed solution, standby; Again cholesterol formyl chloride is dissolved in the chloroformic solution, the concentration of cholesterol formyl chloride is 50～100mg/ml, this solution slowly is added drop-wise in the mixed solution under above-mentioned catalysis and the magnetic agitation, react and stop after 36～54 hours, the molar ratio of the reaction of chitosan and cholesterol formyl chloride is 1～4: 4～1, promptly get the chitin copolymer of modified by cholesterol formyl chloride;
3, the separation and purification of the chitin copolymer of modified by cholesterol formyl chloride:
The chitin copolymer of modified by cholesterol formyl chloride is filtered, filtrate pack in deionized water, dialysed 5～7 days in the bag filter after, the bag filter molecular cut off is 8000～14000, obtain solids, dry, separate in silicagel column, eluant is a dichloromethane or with petroleum ether and ethyl acetate, wherein petroleum ether and ethyl acetate are 4～6: 1 mixed by volume, collect the eluent of product, dichloromethane or petroleum ether and ethyl acetate are removed in volatilization, and drying obtains the pressed powder of the chitin copolymer of modified by cholesterol formyl chloride;
4, the preparation of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride:
With the pressed powder of the chitin copolymer of medicine and modified by cholesterol formyl chloride is that 1: 1～10 ratio is dissolved in the dehydrated alcohol by weight, the pressed powder of the chitin copolymer of medicine and modified by cholesterol formyl chloride is dissolved fully, get lysate, the concentration of medicine in dehydrated alcohol is 0.5～1mg/ml, the chitin copolymer of modified by cholesterol formyl chloride concentration in dehydrated alcohol is 0.5mg/ml～10mg/ml, other removes ionized water, magnetic agitation, the lysate of above-mentioned adding dehydrated alcohol slowly is added drop-wise in this deionized water, the volume ratio of dehydrated alcohol and deionized water is 1: 2～5, after dripping, continue to stir 20min～120min again, then at 20 ℃～70 ℃, 0.4kpa～1kpa decompression, use the Rotary Evaporators rotary evaporation, be evaporated to and stir 1/5～1/3 of back liquor capacity, the solution that the obtains bag filter of packing into is dialysis medium dialysis 50～100min with the deionized water, bag filter molecular cut off 8000～14000, promptly obtain the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride, the drug loading of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride is 2%～15%, and said Rotary Evaporators is the Rotary Evaporators of the model RE-52AA of Shanghai Yarong Biochemical Instrument Plant's production.
The chitosan copolymer carrier micelle of modified by cholesterol formyl chloride can the unfavorable medicine of coated water-soluble, below is example with rubescensine A (model drug), and concrete preparation process is as follows:
1, the purification of chitosan:
Take by weighing the chitosan crude product of 5.00g molecular weight 50kDa, be dissolved in the glacial acetic acid solution of 350ml 0.1M, stirring and dissolving, filter, filtrate adds the NaOH solution of 180ml 0.2M, filter, with the deionized water wash filter cake to pH value 6, place the freezer dryer lyophilization, obtain the chitosan of purification;
2, the preparation of the chitin copolymer of modified by cholesterol formyl chloride and purification:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain, the chitin copolymer that the synthetic cholesterol formyl chloride is modified, wherein, the chitosan 1.008g that gets molecular weight behind the purification earlier and be 50kDa puts in the round-bottomed flask, the solution that adds 50ml dimethyl sulphoxide solution and 27ml chloroform, add triethylamine 2ml and magnetic agitation then, get mixed solution, standby; Get the 1.360g cholesterol formyl chloride, be dissolved in the 27ml chloroformic solution, slowly be added dropwise to then in the mixed solution under above-mentioned catalysis and the magnetic agitation, magnetic agitation reaction 48 hours, the mol ratio of chitosan and cholesterol formyl chloride reaction is 2: 1, takes out reactant liquor, filter, the filtrate bag filter of packing into is the dialysis medium with the deionized water, dialyses 5 days, the bag filter molecular cut off is 8000～14000, take out the solids in the bag filter, separate in silicagel column dry back, and eluant is a dichloromethane, collect the product eluent, fling to dichloromethane, place drying machine dry, promptly obtain the pressed powder of chitin copolymer of the modified by cholesterol formyl chloride of purification;
3, the mensuration of the preparation of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride and physicochemical property:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain; the chitin copolymer that the synthetic cholesterol formyl chloride is modified: the pressed powder 50.8mg of chitin copolymer that takes by weighing the modified by cholesterol formyl chloride of purification; rubescensine A 9.7mg; be dissolved in the 15ml dehydrated alcohol; get lysate; standby; get the 60ml deionized water again; magnetic agitation; the lysate of above-mentioned adding dehydrated alcohol slowly is added drop-wise in this deionized water; drip off the back and continue to stir 30min; then at 40 ℃; 0.7kpa decompression; use the Rotary Evaporators rotary evaporation; be evaporated to 20ml solution; the solution that obtains is packed bag filter into the deionized water 60min that dialyses; the molecular weight that bag filter is held back is 8000～14000; promptly obtain the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride; the drug loading 5.72% of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride; measure through particle size and surface potential detection instrument; the particle diameter of this carrier micelle in water is 70～90nm, surface potential is-31～-30mV, micellar envelop rate is 50%～60%.
Product of the present invention has been obtained good effect through test, and is common with the chitosan of modified by cholesterol formyl chloride. and the polymers carrier micelle act as example to the cancer cell growth inhibited, and interrelated data is as follows:
S grows in the RPMI-1640 culture fluid that contains 10% calf serum.Cell culture condition is: 37 ℃, and 5%CO
2, saturated humidity, every 2-3 days go down to posterity with 0.25% trypsinization.Get cell confession experiment exponential phase of growth.With 0.5 * 10
4/ mL cell inoculation is in 96 well culture plates, every hole 200 μ L, and at 37 ℃, 5%CO
2Cultivate under the condition.Dilute Oridonin-loaded micelle and rubescensine A solution respectively to testing desired concn with the RPMI-1640 culture fluid, after 24 hours, add 96 orifice plates in cell inoculation, every hole 200 μ L.Zeroing group and matched group add the culture fluid of respective volume, establish 4 parallel holes for every group.Cultivate after 48 hours, every hole adds 5mg/mL MTT 20 μ L (except the zeroing group), cultivates 4 hours again, and the culture fluid that inclines adds DMSO 150 μ L/ holes, treats that precipitation fully after the dissolving, reads absorbance (A) with microplate reader after wavelength 570nm place returns to zero.Tumor cell group with not dosing is contrast, calculates IC
50Value.
Under the same experimental conditions, the Hela cell inoculation was cultivated in 96 orifice plates after 24 hours, added Oridonin-loaded micelle and rubescensine A solution through the same concentrations of RPMI-1640 culture fluid dilution, every hole 200 μ L.Zeroing group and matched group add the culture fluid of respective volume, establish 4 parallel holes for every group.Cultivate after 4 hours, withdraw from the culture fluid of pastille, add fresh medium, every hole 200 μ L continue to cultivate after 72 hours, every hole adds 5mg/mL MTT 20 μ L (except the zeroing group), cultivated 4 hours, the culture fluid that inclines adds DMSO150 μ L/ hole again, after treating that precipitation is dissolved fully, after the zeroing of wavelength 570nm place, read absorbance (A) with microplate reader.Tumor cell group with not dosing is contrast, calculates IC
50Value.
Measure Oridonin-loaded micelle and solution inhibitory action with mtt assay to the growth of Hela cell.Oridonin-loaded micelle has obvious suppression and lethal effect to Hela cell growth, and blank micelle is to Hela cell grow almost unrestraint and lethal effect.Along with increasing of Oridonin-loaded micelle concentration, its inhibition degree strengthens, and compares with control formulation rubescensine A solution, and significant difference (p＜0.001) is arranged.Oridonin-loaded micelle acts on the IC of Hela cell
50Value is approximately littler 3 times than rubescensine A solution.Medicine directly acts on 48 hours, the IC of Oridonin-loaded micelle and solution
50Value is respectively 1.25 μ g/mL and 3.44 μ g/mL; Drug effect is changed to fresh medium and cultivated 72 hours after 4 hours, the IC of Oridonin-loaded micelle and solution
50Value is respectively 3.32 μ g/mL and 9.02 μ g/mL.The result shows that Oridonin-loaded micelle obviously is better than rubescensine A solution to the inhibitory action of Hela cell growth.
In sum, the present invention prepares simply, and is effective, makes medicine keep stability, reduces the poison of medicine The property, the growth of establishment cancer cell makes medicine slowly-releasing in vivo, to solve the drug therapy cancer The problem that result for the treatment of is poor during disease, toxic and side effect is big, this invention has huge economic and social benefit.
Claims (3)
1, a kind of preparation method of chitosan copolymer carrier micelle of modified by cholesterol formyl chloride is characterized in that, the chitin copolymer of said modified by cholesterol formyl chloride, and its structural formula is as follows:
Wherein m, n are the degree of polymerization, m+n≤20, and the molecular weight of chitosan is 3～50kDa, and deacetylation is 70～100%, and its preparation method is realized by following steps:
(1), the purification of chitosan: the chitosan crude product of getting molecular weight 3～50kDa, the glacial acetic acid solution that adds 65～85 times 0.1M of chitosan weight, stirring and dissolving is filtered, and adds the sodium hydroxide solution of 0.2M in the filtrate, in and glacial acetic acid, chitosan is precipitated fully, filters, with the deionized water wash filter cake to pH value 67, lyophilization obtains the chitosan of purification;
(2), the preparation of the chitin copolymer of modified by cholesterol formyl chloride:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain, the chitin copolymer that the synthetic cholesterol formyl chloride is modified, step is: the chitosan and the cholesterol formyl chloride that take by weighing purification, chitosan is dispersed in volume ratio: 1～3: in 3～1 the dimethyl sulfoxide and chloroformic solution, the concentration of chitosan is 5～15mg/ml in the solution, add triethylamine 1～2ml and carry out catalysis and magnetic agitation, get mixed solution, standby; Again cholesterol formyl chloride is dissolved in the chloroformic solution, the concentration of cholesterol formyl chloride is 50～100mg/ml, this solution slowly is added drop-wise in the mixed solution under above-mentioned catalysis and the magnetic agitation, reacted 36～54 hours, the molar ratio of the reaction of chitosan and cholesterol formyl chloride is 1～4: 4～1, promptly get the chitin copolymer of modified by cholesterol formyl chloride;
(3), the separation and purification of the chitin copolymer of modified by cholesterol formyl chloride:
The chitin copolymer of modified by cholesterol formyl chloride is filtered, filtrate pack in deionized water, dialysed 5～7 days in the bag filter after, the bag filter molecular cut off is 8000～14000, obtain solids, dry, separate in silicagel column, eluant is the mixed liquor of dichloromethane or petroleum ether and ethyl acetate, wherein petroleum ether and ethyl acetate are 4～6: 1 mixing by volume, collect the eluent of product, dichloromethane or petroleum ether and ethyl acetate are removed in volatilization, and drying obtains the pressed powder of the chitin copolymer of modified by cholesterol formyl chloride;
(4), the preparation of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride:
With the pressed powder of the chitin copolymer of medicine and modified by cholesterol formyl chloride is that 1: 1～10 ratio is dissolved in the dehydrated alcohol by weight, the pressed powder of the chitin copolymer of medicine and modified by cholesterol formyl chloride is dissolved fully, get lysate, the concentration of medicine in dehydrated alcohol is 0.5～1mg/ml, the chitin copolymer of modified by cholesterol formyl chloride concentration in dehydrated alcohol is 0.5～10mg/ml, other removes ionized water, magnetic agitation, the lysate of above-mentioned adding dehydrated alcohol slowly is added drop-wise in this deionized water, the volume ratio of dehydrated alcohol and deionized water is 1: 2～5, after dripping, continue to stir 20～120min again, then at 20 ℃～70 ℃, 0.4～1kpa decompression, use the Rotary Evaporators rotary evaporation, be evaporated to and stir 1/5～1/3 of back liquor capacity, the solution that the obtains bag filter of packing into is dialysis medium dialysis 50～100min with the deionized water, bag filter molecular cut off 8000～14000, promptly obtain the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride, the drug loading of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride is 2%～15%.
2, the preparation method of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride according to claim 1 is characterized in that, is realized by following steps:
(1), the purification of chitosan:
Take by weighing the chitosan crude product of 5.00g molecular weight 50kDa, be dissolved in the glacial acetic acid solution of 350ml 0.1M, stirring and dissolving, filter, filtrate adds the NaOH solution of 180ml 0.2M, filter, with the deionized water wash filter cake to pH value 6, place the freezer dryer lyophilization, obtain the chitosan of purification;
(2), the preparation of the chitin copolymer of modified by cholesterol formyl chloride and purification:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain, the chitin copolymer that the synthetic cholesterol formyl chloride is modified, wherein, the chitosan 1.008g that gets molecular weight behind the purification earlier and be 50kDa puts in the round-bottomed flask, the solution that adds 50ml dimethyl sulphoxide solution and 27ml chloroform, add triethylamine 2ml and magnetic agitation then, get mixed solution, standby; Get the 1.360g cholesterol formyl chloride, be dissolved in the 27ml chloroformic solution, slowly be added dropwise to then in the mixed solution under above-mentioned catalysis and the magnetic agitation, magnetic agitation reaction 48 hours, the mol ratio of chitosan and cholesterol formyl chloride reaction is 2: 1, takes out reactant liquor, filter, the filtrate bag filter of packing into is the dialysis medium with the deionized water, dialyses 5 days, the bag filter molecular cut off is 8000～14000, take out the solids in the bag filter, separate in silicagel column dry back, and eluant is a dichloromethane, collect the product eluent, fling to dichloromethane, place drying machine dry, promptly obtain the pressed powder of chitin copolymer of the modified by cholesterol formyl chloride of purification;
(3), the preparation of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride:
Utilize the amide reaction between the amino deacetylated on the acid chloride groups that has strong reactivity in the cholesterol formyl chloride and the chitosan main chain; the chitin copolymer that the synthetic cholesterol formyl chloride is modified: the pressed powder 50.8mg of chitin copolymer that takes by weighing the modified by cholesterol formyl chloride of purification; rubescensine A 9.7mg; be dissolved in the 15ml dehydrated alcohol; get lysate; standby; get the 60ml deionized water again; magnetic agitation; the lysate of above-mentioned adding dehydrated alcohol slowly is added drop-wise in this deionized water; drip off the back and continue to stir 30min; then at 40 ℃; 0.7kpa the Rotary Evaporators rotary evaporation is used in decompression, is evaporated to 20ml solution; the solution that obtains is packed bag filter into the deionized water 60min that dialyses; the molecular weight that bag filter is held back is 8000～14000, promptly obtains the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride, the drug loading 5.72% of the chitosan copolymer carrier micelle of modified by cholesterol formyl chloride.
3, the chitosan copolymer carrier micelle of claim 1 or 2 described modified by cholesterol formyl chloride is to prepare the application of the medicine that suppresses the cancer cell growth.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101850122A (en) * | 2010-05-21 | 2010-10-06 | 中国医学科学院生物医学工程研究所 | Cholesterylchitosan nano carrier as well as medicament-carried nano particle and preparation method thereof |
| CN104844728A (en) * | 2015-02-02 | 2015-08-19 | 浙江农林大学 | Preparation method and application of cholesterol-carboxymethyl chitosan derivative meterials |
| CN107320447A (en) * | 2017-06-23 | 2017-11-07 | 郑州大学 | A kind of preparation method and application of targeting poly sialic acid mixing nano-micelle |
| CN109879977A (en) * | 2019-01-30 | 2019-06-14 | 中山大学 | A kind of amphiphilic polysaccharide derivative and its preparation method and application containing cholesterol and phytolectin group |
| CN111375067A (en) * | 2020-03-18 | 2020-07-07 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
| CN112341551A (en) * | 2020-11-05 | 2021-02-09 | 广东广纳安疗科技有限公司 | Chitosan substance with ultralow endotoxin and preparation method thereof |
| CN113105566A (en) * | 2021-04-08 | 2021-07-13 | 华中科技大学同济医学院附属协和医院 | Amphiphilic hydroxyethyl starch coupled cholesterol polymer and nano drug-loading system |
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- 2009-09-15 CN CN2009100661516A patent/CN101658484B/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101850122A (en) * | 2010-05-21 | 2010-10-06 | 中国医学科学院生物医学工程研究所 | Cholesterylchitosan nano carrier as well as medicament-carried nano particle and preparation method thereof |
| CN104844728A (en) * | 2015-02-02 | 2015-08-19 | 浙江农林大学 | Preparation method and application of cholesterol-carboxymethyl chitosan derivative meterials |
| CN104844728B (en) * | 2015-02-02 | 2017-04-05 | 浙江农林大学 | A kind of preparation method of cholesterol carboxymethyl chitosan derivative material and application |
| CN107320447A (en) * | 2017-06-23 | 2017-11-07 | 郑州大学 | A kind of preparation method and application of targeting poly sialic acid mixing nano-micelle |
| CN107320447B (en) * | 2017-06-23 | 2020-02-07 | 郑州大学 | Preparation method and application of targeting polysialic acid mixed nano micelle |
| CN109879977A (en) * | 2019-01-30 | 2019-06-14 | 中山大学 | A kind of amphiphilic polysaccharide derivative and its preparation method and application containing cholesterol and phytolectin group |
| CN109879977B (en) * | 2019-01-30 | 2022-03-04 | 中山大学 | Amphiphilic polysaccharide derivative containing cholesterol and phytohemagglutinin groups and preparation method and application thereof |
| CN111375067A (en) * | 2020-03-18 | 2020-07-07 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
| CN111375067B (en) * | 2020-03-18 | 2022-02-11 | 四川大学 | Chitosan scaffold and preparation method and application thereof |
| CN112341551A (en) * | 2020-11-05 | 2021-02-09 | 广东广纳安疗科技有限公司 | Chitosan substance with ultralow endotoxin and preparation method thereof |
| CN113105566A (en) * | 2021-04-08 | 2021-07-13 | 华中科技大学同济医学院附属协和医院 | Amphiphilic hydroxyethyl starch coupled cholesterol polymer and nano drug-loading system |
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