CN103720663A - Follicle-stimulating hormone sustained-release microspheres and preparation method thereof - Google Patents
Follicle-stimulating hormone sustained-release microspheres and preparation method thereof Download PDFInfo
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Abstract
The invention discloses follicle-stimulating hormone sustained-release microspheres. The follicle-stimulating hormone sustained-release microspheres comprise follicle-stimulating hormone, poly lactic-glycolic acid copolymer with the weight-average molecular weight of 7,000 to 25,000, polyethylene glycol and polyvinyl alcohol; the average particle size of the sustained-release microspheres is 8.1 to 9.1 mu m. The sustained-release microspheres disclosed by the invention have the advantages of good shape, uniform particle size distribution, long sustained-release time and high cumulative release rate; the dosing frequency of patients can be reduced; the compliance of the patients is improved. The invention also provides a preparation method of the sustained-release microspheres. The method takes a double-emulsion/ solvent evaporation method as a basic process and has the advantages of high encapsulation efficiency, high medicament loading rate, high microsphere forming rate, high recovery rate, high repeatability and the like.
Description
Technical field
The present invention relates to medical technical field, particularly a kind of follicle stimulating hormone sustained-release micro-spheres and preparation method thereof.
Background technology
In married crowd, have according to the study 15% Mr. and Mrs nearly to exist sterility and infertility, and wherein nearly half is caused by male factor.The reason that causes male sterility is complicated, and wherein azoospermia has accounted for greatly 10%.Azoospermia can be divided into Obstructive azoospermia (Obstructive azoospermia, OA) and non-obstructivity azoospermia (Non-obstructive azoospermia, NOA).NOA patient's testicular spermatogenic function defect, even can not produce sperm.In part NOA patient testis, still can find sperm, by intracytoplasmic sperm injection (ICSI), obtain fertility chance; In another part testis, aspermatogenic patient does not just give birth to chance, and this part patient is the difficult point for the treatment of.Research finds in a lot of testis, do not have sperm NOA patient testis part to have spermatogenic cell, even if only sustenticular cell syndrome (sertoli cell only syndrome, SCOS) also can find spermatogenic cell in the part of its testis; This result of study has been given us useful enlightenment: if can effectively promote that in NOA patient's testis, remaining germ cell developments at different levels become sperm, be expected to obtain the sperm of doing ICSI treatment, solve a clinical difficult problem.The solution of expecting at present has the In vitro culture of allogeneic sexual cell transplanting and sexual cell, but cannot promote due to a variety of causes such as ethics and technology.
Follicle stimulating hormone (follicle stimulating hormone, FSH) is a kind of glycoprotein hormones of hypophysis, and it is essential for the normal reproductive function that maintains masculinity and femininity.In male, FSH has determined quantity and the spermatogenetic quality and quantity of sustenticular cell in testis.Therefore, FSH is considered to impel spermatogenesis and ripe most important hormone.Follicle Stimulating Hormone Receptors (follicle stimulating hormone receptor, FSHR) is expressed on the serous coat of sustenticular cell, and FSH is by working in conjunction with activated G protein-coupled mechanism after FSHR.Fsh receptor signal is essential to stimulating the propagation of sustenticular cell and maintaining the normal sperm generation of testis.The ripe Analytical Chemical Experiment of external sexual cell confirms to increase the FSH concentration in culture fluid, is conducive to the ripe differentiation of sexual cell.In view of above result of study, we expect: increase the concentration of FSH in NOA patient's testis, likely make its remaining germ cell development become sperm, make NOA patient obtain fertility chance.But FSH belongs to polypeptide drug, the short frequent drug administration that needs of protein and peptide class drug half-life maintains effective treatment concentration, causes treatment cycle long, and frequency injection is many, and patient compliance is poor, has adhered to that course for the treatment of person is few.
Therefore, provide a kind of follicle stimulating hormone sustained-release micro-spheres that can continue for a long time to play a role to have important practical significance.
Summary of the invention
The technical problem to be solved in the present invention is for providing a kind of follicle stimulating hormone sustained-release micro-spheres and preparation method thereof.The present invention is take poly lactic coglycolic acid as carrier material, and the follicle stimulating hormone sustained-release micro-spheres that adopts emulsion-solvent evaporation method to prepare can continuous release in body 17 days, and in body, preparation reaches 70%.
Particularly, the invention provides a kind of follicle stimulating hormone sustained-release micro-spheres, comprise follicle stimulating hormone, weight average molecular weight is 7,000-25,000 poly lactic coglycolic acid, cosolvent and stabilizing agent; The mean diameter of described follicle stimulating hormone sustained-release micro-spheres is 8.1～9.1 μ m.
Microsphere sustained-release drug-supplying system is a kind of pharmaceutical carrier drug-supplying system, it take biodegradable polymer as carrier material by drug encapsulation in microsphere supported, by the duct of microsphere inside and the corrosion of the macromolecular material regulating medicine release in vivo and in vitro of degrading, prolong drug action time, improve medicine stability, change medicine distribution in vivo, medicine is concentrated in target area, improve curative effect, reduce toxic and side effects.Carrier material of the present invention is poly lactic coglycolic acid, by two kinds of monomers---lactic acid and hydroxyacetic acid are polymerized at random, be a kind of degradable functional polymer organic compound, there is good biocompatibility, nontoxic, good encystation and the performance of film forming.
As preferably, in described poly lactic coglycolic acid, the polymerization ratio of lactic acid and hydroxyacetic acid (mol ratio) is 50:50.
As preferably, the weight ratio of described follicle stimulating hormone and poly lactic coglycolic acid is 1:2 × 10
5.
As preferably, the molecular weight of described polyvinyl alcohol is 31000-50000.
As preferably, the weight ratio of described follicle stimulating hormone and Polyethylene Glycol, polyvinyl alcohol is 0.00001:0.02:1.
As preferably, cosolvent is a kind of or both the above mixture in Polyethylene Glycol, lactose, microcrystalline Cellulose or polyvinylpyrrolidone.
As preferably, stabilizing agent is polyvinyl alcohol.
Sustained-release micro-spheres form of the present invention is good, particle size distribution is even, and sustained release performance is good, shows following two aspects: (1) slow-release time is long: external slow release can reach one month, in body, discharges sustainable 17 days; (2) preparation is high: cumulative in vitro release rate is about 80%, and in body, preparation is about 85%; Can reduce patient's administration frequency, improve patient's compliance.In addition, follicle stimulating hormone sustained-release micro-spheres drug loading of the present invention can reach 33.7mIU/mg.
The present invention also provides a kind of preparation method of above-mentioned follicle stimulating hormone sustained-release micro-spheres, and the method comprises the following steps:
(1) just the interior aqueous phase solution of 0.2-0.8 parts by volume mixes with 6-10 parts by volume oil-phase solution, and stirs 30s formation colostrum with 4000-6000rpm rotating speed;
In percent weight in volume, described interior aqueous phase solution is that follicle stimulating hormone content is 2 × 10
-3the aqueous solution that %, polyethyleneglycol content are 4%, described oil-phase solution is that poly lactic coglycolic acid content is 25% dichloromethane solution;
The first Ruzhong forming, the weight ratio of follicle stimulating hormone and poly lactic coglycolic acid is 1:2 × 10
5;
(2) 8.5 parts by volume step (1) gained colostrums and the outer aqueous phase solution of 100 parts by volume are mixed, and stir 3-4min formation emulsion with 4000-6000rpm rotating speed;
Described outer aqueous phase solution is that percent weight in volume is 1% polyvinyl alcohol water solution;
(3) step (2) gained emulsion is removed dichloromethane, and centrifugal, collecting precipitation, obtains follicle stimulating hormone sustained-release micro-spheres.
The present invention also provides a kind of follicle stimulating hormone sustained-release micro-spheres being made by above-mentioned preparation method.
The preparation method of sustained-release micro-spheres of the present invention has envelop rate high (can reach 85.5%), drug loading high (can reach 33.7mIU/mg), balling ratio high (can reach 92%), the response rate high (can reach 59.6%), the advantages such as favorable repeatability.
Accompanying drawing explanation
Fig. 1 is perusal follicle stimulating hormone sustained-release micro-spheres finished product;
Fig. 2 is the form of follicle stimulating hormone sustained-release micro-spheres under light microscopic;
Fig. 3 is the form of follicle stimulating hormone sustained-release micro-spheres under Electronic Speculum;
Fig. 4 is the section of follicle stimulating hormone sustained-release micro-spheres under Electronic Speculum;
Fig. 5 is the particle size distribution rectangular histogram of follicle stimulating hormone sustained-release micro-spheres;
Fig. 6 is the cumulative in vitro release profiles of FSH sustained-release micro-spheres of the present invention;
Fig. 7 is the interior plasma concentration curve that discharges of the body of FSH sustained-release micro-spheres of the present invention.
The specific embodiment
In order to make those skilled in the art understand better technical scheme of the present invention, below in conjunction with specific embodiment, the present invention is described in further detail.
Main agents source:
Follicle stimulating hormone (FSH): F4021-10UG, From Human Pituitary, Mw32KDa, Sigma-Aldrich USA;
Polylactic acid-hydroxide acetic acid (PLGA): 719897, Acid Terminated50:50, Mw7000-17000, Sigma-Aldrich, USA;
Polyvinyl alcohol (PVA): 36318, Mw31000-50000, Sigma-Aldrich, USA;
Polyethylene Glycol (PEG): Chemical Reagent Co., Ltd., Sinopharm Group, China.
The preparation of embodiment 1 follicle stimulating hormone sustained-release micro-spheres of the present invention
Preparation technology:
(1) water (W1) in preparation: get 10 μ g FSH, 20mg PEG, adds distilled water to final volume 0.5ml;
(2) preparation oil phase (O): precision takes 2g PLGA(MW:14000), add 8ml dichloromethane fully to dissolve, be made into 25% PLGA solution;
(3) the outer water of preparation (W2): taking PVA(molecular weight is 42000) 1.0g, add a small amount of water for injection make it abundant swelling after, be dissolved in water for injection and be dissolved to 100ml, concentration is 1%.With filter paper filtering, be positioned over 4 ℃ stand-by;
(4) with sample loading gun, W1 is splashed in O, under condition of ice bath, emulsion dispersion machine 5000rpm stir about 30 seconds, forms colostrum;
(5) step (4) gained colostrum is poured in step (3) gained W2, under condition of ice bath, 4000-6000rpm stirs 3-4min, forms emulsion;
(6) motor stirrer 1000rpm, stirs 2-3 hour under ice bath, volatilization CH
2cl
2; Treat that organic solvent waves to the greatest extent, 2000rpm is centrifugal, collects supernatant and preserves, and gets precipitation.Distilled water wash 3 times, recentrifuge is collected, and vacuum lyophilization obtain the follicle stimulating hormone sustained-release micro-spheres of lyophilizing.The preparation of embodiment 2 follicle stimulating hormone sustained-release micro-spheres of the present invention
Preparation technology:
(1) water (W1) in preparation: get 16 μ g FSH, 32mg PEG, adds distilled water to final volume 0.8ml;
(2) preparation oil phase (O): precision takes 2g PLGA(MW:7000), add 10ml dichloromethane fully to dissolve, be made into 20% PLGA solution;
(3) the outer water of preparation (W2): taking PVA(molecular weight is 50000) 1.0g, add a small amount of water for injection make it abundant swelling after, be dissolved in water for injection and be dissolved to 100ml, concentration is 1%.With filter paper filtering, be positioned over 4 ℃ stand-by;
(4) with sample loading gun, W1 is splashed in O, under condition of ice bath, emulsion dispersion machine 5000rpm stir about 30 seconds, forms colostrum;
(5) step (4) gained colostrum is poured in step (3) gained W2, under condition of ice bath, 4000-6000rpm stirs 3-4min, forms emulsion;
(6) motor stirrer 1000rpm, stirs 2-3 hour under ice bath, volatilization CH
2cl
2; Treat that organic solvent waves to the greatest extent, 2000rpm is centrifugal, collects supernatant and preserves, and gets precipitation.Distilled water wash 3 times, recentrifuge is collected, and vacuum lyophilization obtain the follicle stimulating hormone sustained-release micro-spheres of lyophilizing.
The preparation of embodiment 3 follicle stimulating hormone sustained-release micro-spheres of the present invention
Preparation technology:
(1) water (W1) in preparation: get 4 μ g FSH, 8mg PEG, adds distilled water to final volume 0.2ml;
(2) preparation oil phase (O): precision takes 2g PLGA(MW:17000), add 6ml dichloromethane fully to dissolve, be made into 33.3% PLGA solution;
(3) the outer water of preparation (W2): taking PVA(molecular weight is 31000) 1.0g, add a small amount of water for injection make it abundant swelling after, be dissolved in water for injection and be dissolved to 100ml, concentration is 1%.With filter paper filtering, be positioned over 4 ℃ stand-by;
(4) with sample loading gun, W1 is splashed in O, under condition of ice bath, emulsion dispersion machine 5000rpm stir about 30 seconds, forms colostrum;
(5) step (4) gained colostrum is poured in step (3) gained W2, under condition of ice bath, 4000-6000rpm stirs 3-4min, forms emulsion;
(6) motor stirrer 1000rpm, stirs 2-3 hour under ice bath, volatilization CH
2cl
2; Treat that organic solvent waves to the greatest extent, 2000rpm is centrifugal, collects supernatant and preserves, and gets precipitation.Distilled water wash 3 times, recentrifuge is collected, and vacuum lyophilization obtain the follicle stimulating hormone sustained-release micro-spheres of lyophilizing.
The mass parameter evaluation of embodiment 4 follicle stimulating hormone sustained-release micro-spheres of the present invention
(1) morphological feature
As shown in Figure 1, follicle stimulating hormone sustained-release micro-spheres is white in color Powdered perusal result;
(2) the microsphere form under light microscopic
Get lyophilizing above-mentioned microsphere a little, with disperseing containing the normal saline of 0.02% Tween 80, use Nikon optical microscope, under 20 × 10 times of amplifications, observe microsphere form, result as shown in Figure 2, microsphere features smooth surface, scattered, to each other without adhesion.
(3) the microsphere form under Electronic Speculum
Adopt electronic scanner microscope (SEM) to observe microsphere surface and inner exterior appearance and dispersibility, experimental implementation is as follows: the microsphere sample after lyophilizing is fixed on metal spraying on aluminum matter round platform by double faced adhesive tape, under the accelerating potential of JEOLJSM7401F scanning electron microscope 1.0kV, carry out morphologic observation, amplification 250～10,000; As shown in Figure 3, Fig. 3 shows the Electronic Speculum result of follicle stimulating hormone sustained-release micro-spheres of the present invention: microsphere features smooth surface, and scattered, to each other without adhesion; Under Electronic Speculum, microsphere section as shown in Figure 4.
(4) balling ratio:
100 microspheres of random observation under light microscopic, wherein, circular complete microsphere is 92, show that thus balling ratio is 92%.
(5) microsphere average grain diameter and particle size distribution:
The particle diameter of measuring 500 microspheres under optical microscope with micrometer, is a unit every 3 μ m particle diameters, counts respectively the microsphere number of each unit;
A) computing formula of microsphere average grain diameter is:
N in formula
1, n
2... ... ..n
nfor microsphere number, d
1, d
2... ... d
nfor particle diameter;
After testing, thus obtained microsphere mean diameter is 8.6 ± 0.5 μ m.
B) distribution of microspherulite diameter:
Take the percent of microsphere number as vertical coordinate (%), size is that (μ m), draws the rectangular histogram that microspherulite diameter distributes, as shown in Figure 5 to abscissa; Distribution is normal distribution.
(6) the microsphere response rate
Take the weight S of thus obtained microsphere after lyophilizing, ask the ratio of S and the quality of materials P summation of total dosage D and input.Its computing formula is as follows:
After testing and calculate, the response rate of embodiment 1 prepared sustained-release micro-spheres is 59.6%.
(7) microsphere drug loading:
Microsphere molding is also volatilized after dichloromethane, centrifugal and collect microsphere supernatant, measures drug level, obtains to prepare in microsphere process and loses dose, according to following formula, calculates drug loading:
Wherein LC is the english abbreviation of drug loading (Loading Capacity); △ D represents total dosage and loses the poor of dose, i.e. actual content of dispersion in microsphere; S is the microspheres quality of gained.
In microsphere, actual content of dispersion adopts minusing to measure: in preparation process, after organic solvent volatilization completely, collect centrifuged supernatant and with its drug level of chemiluminescence determination, actual drug content equals the difference of content of dispersion in total dosage and supernatant.
Chemiluminescence determination drug level adopts continuous two step enzymes to exempt from method and measures: sample is added to containing being coated with in the paramagnetic particles of the anti-hFSH complex of goat anti-mouse-mice and the reaction tube of proteinaceous TRIS buffer saline.HFSH standing hFSH in solid phase is combined.Be combined in material in solid phase and will be placed in a magnetic field and be held, and unconjugated material is rinsed and removes.Then the anti-hFSH conjugate of alkali phosphatase goat is added and also and had previously been combined in the hFSH combination on microgranule.Separate once again and rinse out unconjugated material.Then chemical luminous substrate Lumi-Phos530 is added in reaction tube, then by illumination meter, the light producing in reaction is measured, the amount of the light that produces is directly proportional to the concentration of hFSH in sample; The drafting of standard curve: with containing 0IU/L, 1.0IU/L, 10IU/L, 50IU/L, 100IU/L, on the titer of 200IU/L, machine is measured, drawing standard curve; Measure as stated above the amount containing FSH light that testing sample produces, according to standard curve, calculate the actual content of FSH.
By said determination method and drug loading computing formula, can be obtained, the drug loading of the prepared follicle stimulating hormone sustained-release micro-spheres of embodiment 1 is 33.7mIU/mg.
(8) microsphere envelop rate
Microsphere envelop rate be in microsphere actual pastille quality and dispensing quality ratio:
Wherein EE is the english abbreviation of envelop rate (Encapsulation efficiency); DT represents total dosage; △ D represents total dosage and loses the poor of dose.
By the computing formula of assay method and the envelop rate of above-mentioned actual content of dispersion, show that the envelop rate of the prepared follicle stimulating hormone sustained-release micro-spheres of embodiment 1 is: 85.5%.
According to above-mentioned test method, detecting envelop rate that embodiment 2 prepares follicle stimulating hormone sustained-release micro-spheres and be the envelop rate that 85.5%, embodiment 3 prepares follicle stimulating hormone sustained-release micro-spheres is 84.9%.
The extracorporeal releasing test of embodiment 5 follicle stimulating hormone sustained-release micro-spheres of the present invention
Grouping: FSH sustained-release micro-spheres (embodiment 1 is prepared) experimental group, blank microsphere matched group, each 6 groups; The making of blank microsphere: replace FSH with mannitol, consumption oozes and is as the criterion with water in the interior water of made and FSH etc., and all the other preparation processes are with embodiment 1.
Extracorporeal releasing test method: precision takes lyophilizing microsphere 20mg, is placed in 1.5ml centrifuge tube, adds the HEPES buffer release medium (10mmol/L PH7.4) of 1.0ml.Sample is put into 37 ℃ of desk-top water bath chaders of constant temperature, and frequency of oscillation is 50rpm, continuous oscillation to the 28 days.Timing fixed point (1d, 3d, 5d, 7d, 9d, 12d, 14d, 18d, 21d, 28d) is taken out sample cell, after 10000rpm frozen centrifugation 10min, takes out supernatant, then adds fresh release medium 1.0ml.Detect the content of FSH in different time points supernatant, and calculate cumulative release degree.In supernatant, the assay of FSH carries out according to the method for chemiluminescence determination drug level described in embodiment 4.
Statistical analysis: the concentration of supernatant FSH represent by average and standard deviation:
experimental group and matched group sample carry out t check analysis, touchstone: α=0.05, with P<0.05, judge there is statistically significant difference.
It is as shown in table 1 that the cumulative in vitro of FSH sustained-release micro-spheres of the present invention discharges measurement result, and its cumulative in vitro release profiles as shown in Figure 6.
Table 1, the external cumulative release measurement result of FSH sustained-release micro-spheres
Table 1 and Fig. 6 result show: the prominent rate of releasing of FSH sustained-release micro-spheres first day of the present invention is 17.2%, enter afterwards steady release period, release can be maintained to the 28th day, and in 28 days, total dose that discharges accounts for 87% of medicine total amount, and within the end one week, have an outburst to discharge, account for 21.8% of total release.
According to above-mentioned test method, the prominent rate of releasing of follicle stimulating hormone sustained-release micro-spheres first day prepared by embodiment 2 is 16.8%, enter afterwards steady release period, release can be maintained to the 28th day, in 28 days, total dose that discharges accounts for 88% of medicine total amount, and within the end one week, have an outburst to discharge, account for 18.2% of total release.
The prominent rate of releasing of follicle stimulating hormone sustained-release micro-spheres first day prepared by embodiment 3 is 18.1%, enter afterwards steady release period, release can be maintained to the 28th day, and in 28 days, total dose that discharges accounts for 86.5% of medicine total amount, and within the end one week, have an outburst to discharge, account for 22.4% of total release.
The vivo releasing test of embodiment 6 follicle stimulating hormone sustained-release micro-spheres of the present invention
Grouping: FSH sustained-release micro-spheres (embodiment 1 is prepared) experimental group, blank microsphere matched group, each 6 groups; The making of blank microsphere is with embodiment 5.
The preparation of injection solvent: 0.3g sodium carboxymethyl cellulose, 0.1g tween 20,0.9g sodium chloride are added and mixes and obtain injection solvent in 10ml distilled water.
Vivo releasing test method: precise electronic balance takes 30mg microsphere, is suspended in 0.5ml injection solvent and is obtained microsphere injection liquid; Get rat, 0.5ml microsphere injection liquid be expelled to rat back leg intramuscular, in 1d, 3d, 5d, 7d, 9d, 12d, 17d, the same time period of 21d (about 8:00-9:00) angular vein clump get blood 3-5ml, 3000rpm centrifuging and taking supernatant, minus 20 degrees preserve to be measured; With chemoluminescence method, measure the concentration of FSH in blood.
Statistical analysis: in blood, the concentration of FSH represent by average and standard deviation:
experimental group and matched group sample carry out t check analysis, touchstone: α=0.05, with P<0.05, judge there is statistically significant difference.
In the body of FSH sustained-release micro-spheres prepared by the embodiment of the present invention 1, discharge determination of plasma concentration result as shown in table 2, in its body, discharge plasma concentration curve as shown in Figure 7.
In table 2, FSH sustained-release microsphere, discharge determination of plasma concentration result
Table 2 and Fig. 7 result show: FSH sustained-release micro-spheres prepared by embodiment 1 can continuous release 17 days, first day has large prominent releasing, discharge subsequently slow decreasing, at the 5th to the 7th day, approach normal level, for discharging lag phase, since the 8th day concentration rising again, at fortnight, again reach peak value, by the 17 day, again approach normal level; In 17 days endosomes, preparation is about 85%.
The follicle stimulating hormone sustained-release micro-spheres that according to the method described above prepared by the embodiment of the present invention 2 can continuous release 18 days, first day has large prominent releasing, discharge subsequently slow decreasing, at the 4th to the 6th day, approach normal level, for discharging lag phase, since the 7th day concentration rising again, at the 13 day, again reach peak value, by the 15 day, again approach normal level; In 18 days endosomes, preparation is about 87.4%.
The follicle stimulating hormone sustained-release micro-spheres that according to the method described above prepared by the embodiment of the present invention 3 can continuous release 16 days, first day has large prominent releasing, discharge subsequently slow decreasing, at the 6th to the 8th day, approach normal level, for discharging lag phase, since the 9th day concentration rising again, at the 15 day, again reach peak value, by the 16 day, again approach normal level; In 16 days endosomes, preparation is about 84.6%.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (9)
1. a follicle stimulating hormone sustained-release micro-spheres, it is characterized in that, comprise follicle stimulating hormone, weight average molecular weight is 7,000-25, the mean diameter of follicle stimulating hormone sustained-release micro-spheres is 8.1～9.1 μ m described in 000 poly lactic coglycolic acid, cosolvent and stabilizing agent.
2. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, in described poly lactic coglycolic acid, the polymerization ratio of lactic acid and hydroxyacetic acid is 50:50.
3. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, the weight ratio of described follicle stimulating hormone and poly lactic coglycolic acid is 1:2 × 10
5.
4. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, the molecular weight of described polyvinyl alcohol is 31000-50000.
5. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, the weight ratio of follicle stimulating hormone and Polyethylene Glycol, polyvinyl alcohol is 0.00001:0.02:1.
6. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, described cosolvent is a kind of or both the above mixture in Polyethylene Glycol, lactose, microcrystalline Cellulose or polyvinylpyrrolidone.
7. follicle stimulating hormone sustained-release micro-spheres as claimed in claim 1, is characterized in that, described stabilizing agent is polyvinyl alcohol.
8. a preparation method for follicle stimulating hormone sustained-release micro-spheres as described in claim 1-7 any one, is characterized in that, comprises the following steps:
(1) just the interior aqueous phase solution of 0.2-0.8 parts by volume mixes with 6-10 parts by volume oil-phase solution, and stirs 30s formation colostrum with 4000-6000rpm rotating speed;
In percent weight in volume, described interior aqueous phase solution is that follicle stimulating hormone content is 2 × 10
-3the aqueous solution that %, polyethyleneglycol content are 4%, described oil-phase solution is that poly lactic coglycolic acid content is 25% dichloromethane solution;
(2) 8.5 parts by volume step (1) gained colostrums and the outer aqueous phase solution of 100 parts by volume are mixed, and stir 3-4min formation emulsion with 4000-6000rpm rotating speed;
Described outer aqueous phase solution is that percent weight in volume is 1% polyvinyl alcohol water solution;
(3) step (2) gained emulsion is removed dichloromethane, and centrifugal, collecting precipitation, obtains follicle stimulating hormone sustained-release micro-spheres.
9. the follicle stimulating hormone sustained-release micro-spheres that preparation method according to claim 8 makes.
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| CN105726468A (en) * | 2016-02-28 | 2016-07-06 | 中国农业科学院特产研究所 | Superovulation polyvinylpyrrolidone FSH composite slow-release injection for sika deer |
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