CN101461786A - PLA/PLGA shell-core microballoons prepared by oil in water-solid in oil method, and preparation method thereof - Google Patents

Abstract

Translated fromChinese

本发明涉及的是一种制药技术领域的用水包油—油包固体制备的PLA/PLGA壳—核微球及其制备方法，该微球包括药物、聚乳酸和/或聚乳酸-羟基乙酸、或/和药用辅料，重量百分比含量为：药物0.01－50％、聚乳酸和/或聚乳酸－羟基乙酸20－99.99％、药用辅料为0－30％。本发明将药物颗粒加到PLA和/或PLGA有机溶液中进行乳化，然后选择亲水性的有机溶剂再次乳化形成未硬化的球，最后在另一大油相硬化，除去有机溶剂并收集微球。本发明避免用常规的W/O和W/O/W的包封率不高，和S/O/O的突释严重，及造成的环境污染的缺点，微球粒径根据需要进行控制，不污染环境，可以运用到各种药物缓释或控释微球的制备及疫苗的佐剂。The present invention relates to PLA/PLGA shell-core microspheres prepared from oil-in-water-solid-in-oil in the technical field of pharmacy and a preparation method thereof. The microspheres include drugs, polylactic acid and/or polylactic acid-glycolic acid, Or/and pharmaceutical auxiliary materials, the weight percentage content is: 0.01-50% of medicine, 20-99.99% of polylactic acid and/or polylactic acid-glycolic acid, and 0-30% of pharmaceutical auxiliary materials. In the present invention, the drug particles are added to the PLA and/or PLGA organic solution for emulsification, and then a hydrophilic organic solvent is selected to emulsify again to form unhardened balls, and finally hardened in another large oil phase, the organic solvent is removed and the microspheres are collected . The present invention avoids the shortcomings of conventional W/O and W/O/W with low encapsulation efficiency, severe sudden release of S/O/O, and environmental pollution, and the particle size of the microspheres is controlled according to needs. It does not pollute the environment, and can be applied to the preparation of various drug slow-release or controlled-release microspheres and vaccine adjuvants.

Description

Wrap PLA/PLGA shell-nuclear microsphere of solid preparation and preparation method thereof with oil-in-water-oil

Technical field

What the present invention relates to is the preparation shell-nuclear microsphere in a kind of pharmaceutical technology field and preparation method thereof, and especially a kind of oil-in-water-oil bag solid (S-O-W) of using prepares the method for PLA (polylactic acid)/PLGA (polylactic acid-hydroxyacetic acid) shell-nuclear microsphere.

Background technology

Pharmaceutical industry is from drug discovery, and to Clinical Application, last link is pharmaceutical preparation.Wherein some medicine needs long term administration to cure; Some needs topicals such as targeting.Reach these purposes, crude drug must be prepared into corresponding dosage forms.For example need long term administration but short medicine of in vivo half-life should be prepared into the PLA dosage form; For some tumor treatment, need some drug targetings in the disease photograph, for example targeting is in tumor vascular embolism microball preparation etc.; Gene recombination technology be used for the treatment of proteic expression and production 20 for many years, up to the present, existing more than 30 protein drug product drops into clinical use, nearly 200 examine with R﹠D process in, emerge and a collection ofly enter (Amgen), gene technology a collection of new large-scale medical companies such as (Genentech) such as peace.With respect to the fast development of protein macromolecule medicine itself, its dosage form technical progress is slow.On the one hand, the protein macromolecule drug oral does not absorb, the interior half-life of body is short, needs drug administration by injection; On the other hand, the protein drug treatment cycle of many He Ermeng, cytokine class is long, and injection for a long time and continually becomes necessary, also influences the main cause of patient's compliance.The research and development of the dosage form of slow release protein drug are owing to cause the easily prominent shortcoming such as release of the microsphere of active loss such as W/O/W method preparation in preparation microgranule process.It is imperative that the protein microsphere that development preparation has an active protection can improve envelop rate again and reduce the prominent method of releasing.People have studied the double-deck microsphere of preparation in order to reduce prominent releasing, but still can not solve this difficult problem up till now.

Find by prior art documents, " Double walled PLA/PLGAmicrospheres:encapsulation of water-soluble and water-insoluble proteinsand their release properties " " Journal of Controlled Release " 89 (2003) 167-177 that Meng Shi etc. delivers], [Meng Shi etc., double-deck POE/PLGA microsphere: seal water solublity and water-insoluble protein and their release feature, the sustained release magazine, 89 (2003) 167-177], people such as Meng Shi have reported in the document and have utilized W/O/W method being encapsulated in PLGA/PLA shell-nuclear microsphere bovine serum albumin (BSA) and Ciclosporin A (CyA).The multi-emulsion method that the document utilizes W/O/W to see most prepares double-deck microsphere, and the oil-water interfaces of multi-emulsion method are the albumen killers who generally acknowledges, causes the gathering of water miscible albumen at this interface, causes envelop rate also not high equally, exists not exclusively to discharge and prominent releasing.Morita T. etc. were " Journal of Controlled Release " (sustained release magazine) (2000, the 69th phase, the 435th page to the 444th page) on deliver " Proteinencapsulation into biodegradable microspheres by a novel S/O/W emulsionmethod using poly (ethylene glycol) as a protein micronization adjuvant " (with Polyethylene Glycol as the micronized excipient of albumen (surfactant), use oil-in-water-solid method of oil bag that the albumen microcapsule is wrapped in the biodegradable microsphere then), the document has been reported and has been utilized new S/O/W emulsion process preparation to carry protein microsphere.Just changed surfactant in the past report more be use PVA, use PEG instead at this piece document.But still it is low to overcome envelop rate, has the prominent shortcoming of releasing and not exclusively discharging.

Summary of the invention

The objective of the invention is to overcome deficiency of the prior art, a kind of PLA/PLGA shell-nuclear microsphere that wraps the solid preparation with oil-in-water-oil and preparation method thereof is provided, make the smooth rounding of microparticle surfaces of its preparation, good evenness, granule regular without adhesion; The envelop rate height, prominent release little, the drug loading height.

The present invention is achieved by the following technical solutions:

PLA/PLGA shell-nuclear microsphere with oil-in-water-oil bag solid preparation involved in the present invention, comprise medicine, polylactic acid and/or polylactic acid-hydroxyacetic acid or/and pharmaceutic adjuvant, their weight percent content is: medicine 0.01-50%, polylactic acid and/or polylactic acid-hydroxyacetic acid 20-99.99%, pharmaceutic adjuvant are 0-30%.This microsphere drug loading is controlled, envelop rate is high, prominent release and not exclusively release.

Described medicine comprises small-molecule drug and macromolecular drug.What described small-molecule drug referred to mainly is chemicals, and macromolecular drug mainly is meant biopharmaceutical macromolecular drug, especially finger protein macromolecular drug, vaccine, antibody, nucleic acid or liposome medicament.

Described protein macromolecule medicine is: erythropoietin (EPO), recombinant human granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimutaing factor (GM-CSF), vaccine, interferon (IFN), growth hormone (GH), insulin (Insulin), epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-β), insulin like growth factor (IGF), vascular endothelial cell growth factor (VEGF), PDGF (PDGF), endothelial cell growth factor (ECGF) (EGF), nerve growth factor (NGF), bone-derived growth factor (BDGF), bone morphogenetic protein(BMP) (BMP), tissue polypeptide antigen (TPA), antibody (antibody), blood coagulation factor VIII (VIII), or plasma thromboplastin component genetic factor.

Described nucleic acid is: antisense nucleotide (anti-RNA), microRNA (RNAi) or gene (DNA).

Described adjuvant refers to mainly is can the injection rank, especially little saccharide (sucrose, trehalose, glucose, maltose, or lactose), polyhydroxy compounds (mannitol, sorbitol, glycerol, 1,2-propylene glycol, erythritol, Polyethylene Glycol, polyvinyl alcohol, poly(ethylene oxide), or polypyrrole alkane ketone), polysaccharide compound (glucosan, sodium alginate, chitosan, starch, cellulose, or cyclodextrin material), amino-acid compound (glycine, lysine, arginine, glutamic acid or histidine), or inorganic salts material (zinc salt, calcium salt, mantoquita, magnesium salt, or molybdenum salt) a kind of or combination in any.

Preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares involved in the present invention is that drug particles is dispersed in the organic solution of PLA and/or PLGA, and the formation suspensions that make it to be uniformly dispersed such as stirring or whirlpool; Then suspension is added to outer oil phase, restir or whirlpool form microsphere, at last it are transferred to big aqueous phase and solidify; Centrifugal then collection microsphere, lyophilizing is preserved.

Said method of the present invention specifically may further comprise the steps:

1. medicine or medicine and pharmaceutic adjuvant are prepared into drug particles;

2. drug particles is joined the organic solution of PLA and PLGA or the organic solution of hydrophilic drug particles and PLGA mixed after, mixed with the organic solution of PLA again; Or it is mixed earlier hydrophobic drug particles and PLA, mixed with the organic solution of PLGA again; Form suspension, i.e. oil phase-1 (O₁) in stir or whirlpool etc. makes it homodisperse and forms uniform suspension;

3. 2. completing steps is formed the aqueous solution emulsifying that microsphere suspension is added drop-wise to sodium chloride solution and surfactant, form emulsion;

4. then step emulsion 3. being transferred to sodium chloride solution solidifies;

5. completing steps sample lyophilizing is 4. obtained PLA/PLGA shell-nuclear microsphere.

Described drug particles refers to hydrophilic drug particles, or medicine is loaded into the granule for preparing in the adjuvant, or the method for hydrophobic medicine by preparation is prepared into the granule that is insoluble to organic solvent;

Described drug particles, the size of its drug particles be at 0.2-10 μ mm, preferred 0.5-5 μ mm.

Described drug particles, it accounts for the 0.2-50% of drug particles gross weight; Preferred 1-20% (w/w).

Described oil phase-(O) be: the organic solution of PLA and/or PLGA;

The organic solution of described PLA and/or PLGA is: the organic solution of PLA and PLGA is added 0.1-20% (w/w) Polyethylene Glycol (PEG) or poloxamer (poloxmer); The dichloromethane of PLA and PLGA, ethyl acetate, acetonitrile, heptane, chloroform or acetone organic solution are good with a kind of in dichloromethane, ethyl acetate or the acetonitrile or their organic solution of combination in any;

The organic solution concentration of described PLA and/or PLGA is: the percent concentration that the weight of PLA accounts for total oil phase is 0.5-80% (w/w), is good with 5-30% (w/w) wherein; Hundred percent concentrations of the total oil phase of weight of PLGA are 20-99.5% (w/w), are good with 70-95% wherein;

Described surfactant is: polyvinyl alcohol (PVA), Polyethylene Glycol (PEG), polyvinylpyrrolidone (PVP), poloxamer (poloxmer), poly-sorbic alcohol, ethyl cellulose (EC) or tween;

The concentration of described surface active agent polyvinyl alcohol (PVA) is 0.5-10% (w/w) or contains saline solution such as 0.5-10% (w/w) sodium chloride, the concentration of Polyethylene Glycol (PEG) is 0.5-20% (w/w) or the saline solution that contains 0.5-10% (w/w) sodium chloride, polyvinylpyrrolidone (PVP) concentration is 0.5-20% (w/w) or contains saline solution such as 0.5-10% (w/w) sodium chloride, poloxamer (poloxmer) concentration is 0.5-20% (w/w) or contains saline solution such as 0.5-10% (w/w) sodium chloride, poly-sorbic alcohol concentration is 0.5-20% (w/w) or contains saline solution such as 0.5-10% (w/w) sodium chloride, or ethyl cellulose (EC) concentration is 0.5-20% (w/w) or contains saline solution such as 0.5-10% (w/w) sodium chloride;

The particle diameter of described microsphere is 1-500 μ m, is good with 10-100 μ m.

The present invention has selected the PLGA of proper ratio and concentration and the composite material of PLA, make water miscible drug particles or oil-soluble medicine be prepared into oily insoluble granule by the method for preparation, avoid not high with the envelop rate of conventional W/O and W/O/W, and the prominent of S/O/O released seriously the shortcoming of the environmental pollution that causes; Adopt this method to prepare microsphere, the size of its particle diameter can be controlled according to different needs, and is free from environmental pollution; Can avoid the function influence to the treatment of medicine, especially those physicochemical properties are unsettled, to the medicine of oil-water interfaces sensitivity, as biopharmaceutical macromolecular drug such as protein macromolecule medicine, DNA, RNA or siRNA medicine.The smooth surface rounding of microgranule, the granule regular without adhesion, particle diameter can be regulated and control from 1 μ m as required to 500 μ m, and its freeze dried powder is that white is fine and smooth, loose, can not subside, adhesion, and redispersibility is good.Can apply to the preparation of various medicament slow releases or control-release microsphere and the adjuvant of vaccine.

The specific embodiment

Below embodiments of the invention are elaborated: present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.

Adopt the inventive method to prepare the microsphere of medicine carrying, when wherein preparing drug particles, to small-molecule drug can be conventional grinding method, the sedimentation method, comminuting method or comminution granulation; For biopharmaceutical macromolecular drug can be the method for the complex of water-aqueous phase emulsion method, low temperature spray drying method, phase separation method, supercritical methanol technology or metal ion and biomacromolecule formation.

Embodiment one: be loaded with the preparation of small-molecule drug granule PLA/PLGA shell-nuclear microsphere

(1) small-molecule drug or small-molecule drug and pharmaceutic adjuvant are prepared into drug particles;

In the present embodiment, the small-molecule drug of employing is that (tumor chemotherapeutic drug is selected from chemotherapy of tumors class medicine: amycin, cyclophosphamide, dactinomycin, bleomycin, daunorubicin, amycin, epirubicin, mitomycin, methotrexate, fluorouracil, carboplatin, carmustine (BCNU), semustine, cisplatin, etoposide, camptothecine and derivant thereof, phenesterin, paclitaxel and derivant thereof, Docetaxel and derivant thereof, vinblastine, vincristine, zitazonium, etoposide, piposulfan, cyclophosphamide or flutamide and derivant thereof; Sustained-release micro-spheres can be loaded with one or more in the said medicine; ) or antibiotics (antibiotics is selected from ciclosporin, levofloxacin, ofloxacin or epinastine hydrochloride; Sustained-release micro-spheres can be loaded with one or more in the said medicine).

In the present embodiment, the pharmaceutic adjuvant of employing is one or more in glucosan, sodium alginate, chitosan, starch, cellulose, cyclodextrin, Polyethylene Glycol, polyvinyl alcohol, poly(ethylene oxide), polypyrrole alkane ketone, sucrose, trehalose, mannitol, sorbitol, lactose, glycine, lysine, zinc salt, calcium salt, mantoquita, magnesium salt, molybdenum salt or the histidine said medicine.

In the present embodiment, concrete proportioning is as follows:

Medicine

Adjuvant

Medicine accounts for the percentage by weight of drug particles

Medicine

Adjuvant

Medicine accounts for the percentage by weight of drug particles

Amycin	Cyclodextrin	100%, 80%, 50 %, 25% or 0%	Carboplatin	Cyclodextrin	100%, 80%, 50 %, 25% or 0%
						Amycin	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%	Carmustine	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%
Amycin	Polyvinyl alcohol	100%, 80%, 50 %, 25% or 0%	Semustine	Polyvinyl alcohol	100%, 80%, 50 %, 25% or 0%
						Amycin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%	Cisplatin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%
Amycin	Polypyrrole alkane ketone	100%, 80%, 50 %, 25% or 0%	Etoposide	Polypyrrole alkane ketone	100%, 80%, 50 %, 25% or 0%
						Cyclophosphamide	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%	Camptothecine	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%
Dactinomycin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%	Phenesterin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%
						Daunorubicin	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%	Paclitaxel	Polyethylene Glycol	100%, 80%, 50 %, 25% or 10%
Amycin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%	Docetaxel	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%
						Epirubicin	Polypyrrole alkane ketone	L00%, 80%, 50 %, 25% or 0%	Vinblastine	Polypyrrole alkane ketone	L00%, 80%, 50 %, 25% or 0%
Methotrexate	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%	Vincristine	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%
						Fluorouracil	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%	Zitazonium	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%
Etoposide	Cyclodextrin	100%, 80%, 50 %, 25% or 0%	Piposulfan	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%
						Flutamide	Polyethylene Glycol	100%, 80%, 50 %, 25% or 0%	Daunorubicin and carboplatin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%
Amycin and paclitaxel	Polyvinyl alcohol	100%, 80%, 50 %, 25% or 0%	Ciclosporin	Polypyrrole alkane ketone	100%, 80%, 50 %, 25% or 0%
						Ciclosporin	Poly(ethylene oxide)	100%, 80%, 50 %, 25% or 0%	Levofloxacin	Poly(ethylene oxide)	100%, 90%, 80 %, 40% or 20%
Levofloxacin	Polypyrrole alkane ketone	100%, 80%, 40 %, 20% or 0%	Ofloxacin	Polyethylene Glycol	100%, 80%, 40 %, 20% or 0%
						Ofloxacin	Glucosan	100%, 80%, 40 %, 10% or 0%	Epinastine hydrochloride	Poly(ethylene oxide)	100％、80％、40 ％、20％、0.1％
Epinastine hydrochloride	Sodium alginate	100%, 80%, 40 %, 20% or 0%	Levofloxacin	Mannitol	100%, 80%, 40 %, 20% or 0%
						Levofloxacin	Chitosan	100%, 80%, 40 %, 20% or 0%	Ofloxacin	Sorbitol	100%, 80%, 40 %, 20% or 0%

Ofloxacin	Starch	100％、80％、40 ％、20％、0.1％	Epinastine hydrochloride	Lactose	100%, 80%, 40 %, 20% or 0%
						Epinastine hydrochloride	Cellulose	100%, 80%, 40 %, 20% or 0%	Levofloxacin	Glycine	100%, 80%, 40 %, 20% or 0%
Ofloxacin	Sucrose	100%, 80%, 40 %, 20% or 0%	Ofloxacin	Lysine	100%, 80%, 40 %, 20% or 0%
						Epinastine hydrochloride	Trehalose	100%, 80%, 40 %, 20% or 0%	Epinastine hydrochloride	Histidine	100%, 80%, 40 %, 20% or 0%

(2) dichloromethane, acetonitrile or the ethyl acetate solution weight ratio according to small-molecule drug granule and PLGA is 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. oil bag solid (S/O) emulsion; Or be the dichloromethane of drug molecule granule and PLA, acetonitrile or ethyl acetate solution weight ratio 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed even suspension in ultrasonic 1-5 minutes, again the organic solution of PLGA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. oil bag solid (S/O) emulsion;

(3) step (2) being got emulsion droplets is added to aqueous surfactant solution and stirring, the whirlpool of 1-10% (w/w) sodium chloride and 1-10% (w/w) or formed emulsion in ultrasonic 0.1-5 minutes;

(4) emulsion of step (3) being added drop-wise to concentration is that the 1000ml sodium chloride solution of 1-10% (w/w) solidified 1-4 hours;

(5) the centrifugal collection microsphere that step (4) is obtained, and wash with water 3-5 times, microsphere obtained after the lyophilizing.

The microsphere that present embodiment obtains, concrete set of dispense ratio sees the following form:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere	Drug particles	The weight ratio of PLGA/PLA	Drug particles account for microsphere percentage by weight
						The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The carboplatin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The carmustine granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The semustine granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80%, 70% or 99.99%

The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80%, 70% or 99.99%	The cisplatin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 50% or 99.99%
						The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 50% or 99.99%	The etoposide granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 50%, 80%, 70% or 99.99%
The cyclophosphamide granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 50%, 80%, 70% or 99.99%	The camptothecine granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The dactinomycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The phenesterin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40%, 70% or 99.99%
The daunorubicin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40%, 70% or 99.99%	The paclitaxel granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The amycin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The Docetaxel granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The epirubicin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The vinblastine granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The methotrexate granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The vincristine granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The fluorouracil granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The zitazonium granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The etoposide granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The piposulfan granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The flutamide granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	Daunorubicin and carboplatin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						Amycin and paclitaxel granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The ciclosporin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80%, 70% or 99.99%
The ciclosporin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The levofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 50% or 99.99%
						The levofloxacin granule	1、1/2、1/3、 1/4、1/5、2、	20%, 60%, 80%, 70% or 99.99%	The ofloxacin granule	1、1/2、1/3、 1/4、1/5、2、	20％、50％、 80％、70％、

	3,4 or 5			3,4 or 5	Or 99.99%
						The ofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The levofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40%, 70% or 99.99%
						The levofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The ofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The ofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 78%, 70% or 99.99%	The levofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
The ofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The ofloxacin granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%
						The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%	The epinastine hydrochloride granule	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The microsphere of this method preparation need can be used to the disease of long-term treatment, especially needs the blood vessel embolism microsphere of the disease such as the tumor of topical therapeutic.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.

Embodiment two: be loaded with the preparation of biopharmaceutical macromolecular drug granule PLA/PLGA shell-nuclear microsphere

(1) biopharmaceutical macromolecular drug or biopharmaceutical macromolecular drug and pharmaceutic adjuvant are prepared into drug particles;

In the present embodiment, the biopharmaceutical macromolecular drug of employing is erythropoietin (EPO), recombinant human granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimutaing factor (GM-CSF), vaccine, interferon (INF), growth hormone (GH), insulin (Insulin), epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF-β), insulin like growth factor (IGF), vascular endothelial cell growth factor (VEGF), PDGF (PDGF), endothelial cell growth factor (ECGF) (ECGF), nerve growth factor (NGF), bone-derived growth factor (BDGF), bone morphogenetic protein(BMP) (BMP), tissue polypeptide antigen (TPA), antibody (antibody), blood coagulation factor VIII (VIII), the IX genetic factor, antisense nucleotide (anti-RNA), microRNA (RNAi), SiRNA, gene (DNA); Sustained-release micro-spheres can be loaded with one or more in the said medicine.

In the present embodiment, the pharmaceutic adjuvant of employing is one or more in glucosan, sodium alginate, chitosan, starch, cellulose, cyclodextrin, Polyethylene Glycol, polyvinyl alcohol, poly(ethylene oxide), polypyrrole alkane ketone, sucrose, trehalose, mannitol, sorbitol, lactose, glycine, lysine or the histidine said medicine;

Composition proportioning concrete in the present embodiment is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight
						EPO	Glucosan	80%, 50%, 25 %, 10%, 5% or 0%	NGF	Cyclodextrin and sucrose (5:1)	100%, 80%, 50 %, 25%, 10%, 5% or 0%
G-CSF	Glucosan and trehalose (9: 1)	80%, 50%, 25 %, 10%, 5% or 0%	BDGF	Polyethylene Glycol and sorbitol (8:1)	100%, 80%, 50 %, 25%, 10%, 5% or 0%
						GM-CSF	Glucosan and trehalose (9: 1)	80%, 50%, 25 %, 10%, 5% or 0%	BMP	Glucosan	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
Vaccine	Glucosan	100%, 80%, 50 %, 25%, 10%, 5% or 0%	TPA	Poly(ethylene oxide) and sucrose (9:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						GH	Glucosan zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 %10%, 5% or 0%	antibody	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
Insulin	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25%, 10%, 5% or 0%	Blood coagulation factor VIII	Glucosan and lysine (9:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						EGF	Poly(ethylene oxide)	100%, 80%, 50 %, 25%, 10%, 5% or 0%	The IX genetic factor	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
FGF	Polyethylene Glycol	100%, 80%, 50 %, 25%, 10%, 5% or 0%	anti-RNA	Polyethylene Glycol	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						TGF-β	Poly(ethylene oxide)	100%, 80%, 50 %, 25%, 10%, 5% or 0%	RNAi	Poly(ethylene oxide)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
IGF	Glucosan	100％、80％、50 ％、25％、10％、	SiRNA	Glucosan	50％、40％、30 ％、25％、15％、

		5% or 0%			10%, 5% or 0%
						VEGF	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%	DNA	Glucosan	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
PDGF	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%	BDGF and BMP (1:1)	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						ECGF	Cyclodextrin and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%	PDGF and BMP (1:1)	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10% or 5%
VEGF and DNA (1:1)	Glucosan and trehalose (8:1)	80%, 70%, 50 %, 25%, 10%, 5% or 0%	TGF-β, PDGF and BMP (1:1:1)	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10% or 5%
						NGF and BM P (1:1)	Glucosan and lactose (8:1)	80%, 60%, 50 %, 25%, 10%, 5% or 0%	SiRNA and DNA (1:1)	Polypyrrole alkane ketone	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
EGF and FG F (1:1)	Glucosan and trehalose (8:1)	60%, 50%, 25 %, 10%, 5% or 0%	BDGF and VEGF (1:1)	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						GM-CSF and G-CSF (1:1)	Glucosan and lactose (8:1)	60%, 50%, 25 %, 10%, 5% or 0%	IGF and GH (1:1)	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
VEGF and DNA (1:1)	Glucosan and sodium alginate (9:1)	60%, 50%, 25 %, 10%, 5% or 0%	EPO	Glucosan and sodium alginate (9:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						NGF and BM P (1:1)	Glucosan and sodium alginate (9:1)	60%, 50%, 40 %, 20%, 10%, 5% or 0%	G-CSF	Glucosan and sodium alginate (9:1)	50%, 40%, 30 %, 25%, 15%, 10% or 5%
EGF and FG F (1:1)	Chitosan	60%, 50%, 40 %, 20%, 10%, 5% or 0%	GM-CSF	Glucosan and sorbitol (9:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						GM-CSF and G-CSF (1:1)	Starch	60%, 50%, 40 %, 20%, 10%, 5% or 0%	Vaccine	Glucosan and lactose (9:1)	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
BDGF	Cellulose and lactose (9:1)	100%, 80%, 40 %, 20%, 10%, 5% or 0%	EPO	Glucosan and glycine	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
						BMP	Glucosan and sucrose (9:1)	100%, 80%, 40 %, 20%, 10%, 5% or 0%	G-CSF	Glucosan and lysine	50%, 40%, 30 %, 25%, 15%, 10%, 5% or 0%
TPA	Trehalose	100％、80％、40	GM-CSF	Glucosan and group	50％、40％、30

%, 20%, 10%, 5% or 0%

Propylhomoserin

%, 25%, 15%, 10%, 5% or 0%

(2) dichloromethane, acetonitrile, ethyl acetate or their the mixed solution weight ratio according to biopharmaceutical macromolecular drug granule and PLGA is 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere	Drug particles	The weight ratio of PLGA/PLA	Medicine accounts for the percentage by weight of microsphere
						EPO	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	NGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
G-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	BDGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						GM-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	BMP	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80 %, 70% or 99.99%
Vaccine	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80 %, 70% or 99.99%	TPA	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 50% or 99.99%
						GH	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 50% or 99.99%	antibody	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 50%, 80 %, 70% or 99.99%
Insulin	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 50%, 80 %, 70% or 99.99%	Blood coagulation factor VIII	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						EGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	The IX genetic factor	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40 %, 70% or 99.99%
FGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40 %, 70% or 99.99%	anti-RNA	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						TGF-β	1、1/2、1/3、	20％、60％、80	RNAi	1、1/2、1/3、	20％、60％、80

	1/4,1/5,2,3,4 or 5	%, 70% or 99.99%		1/4,1/5,2,3,4 or 5	%, 70% or 99.99%
						IGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	SiRNA	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
VEGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	DNA	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						PDGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	BDGF and BMP (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
ECGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	PDGF and BMP (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						VEGF and DNA (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	TGF-β, PDGF and BMP (1:1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
NGF and BM P (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	SiRNA and DNA (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 40%, 80 %, 70% or 99.99%
						EGF and FG F (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	BDGF and VEGF (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 50% or 99.99%
GM-CSF and G-CSF (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	IGF and GH (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 50%, 80 %, 70% or 99.99%
						VEGF and DNA (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	EPO	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
NGF and BM P (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	G-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 40 %, 70% or 99.99%
						EGF and FG F (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	GM-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
GM-CSF and G-CSF (1:1)	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	Vaccine	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
						BDGF	1、1/2、1/3、 1/4、1/5、2、3、	20%, 60%, 78 %, 70% or	EPO	1、1/2、1/3、 1/4、1/5、2、3、	20%, 60%, 80 %, 70% or

	4 or 5	99.99％		4 or 5	99.99％
						BMP	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	G-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%
TPA	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%	GM-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80 %, 70% or 99.99%

The microsphere of this method preparation can be used for the disease of the frequent drug administration by injection of needs, long-term treatment, especially needs the treatment of major diseases such as anemia that the disease of topical therapeutic such as tumor, the various cause of disease cause, dwarf.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously biopharmaceutical macromolecular drug is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment three: be loaded with the preparation of interferon (IFN) drug microparticles PLA/PLGA shell-nuclear microsphere

(1) interferon and pharmaceutic adjuvant are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						IFN	Glucosan	80%, 50%, 25 %, 10% or 5%	IFN	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50 %, 25% or 10%
IFN	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	IFN	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50 %, 25% or 10%
						IFN	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	IFN	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10%	IFN	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	IFN	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	IFN	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%

IFN	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	IFN	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10%	IFN	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10%	IFN	Poly(ethylene oxide)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10%	IFN	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	IFN	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	IFN	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15%10 % or 5%	IFN	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10%	IFN	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50 %, 25% or 10%	IFN	Polypyrrole alkane ketone	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	IFN	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	IFN	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	IFN	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	IFN	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Chitosan and sucrose (9:1) or 6:	60%, 50%, 40 %, 20%, 10% or	IFN	Glucosan and sorbitol (9:1), (7:1), (6:1),	50％、40％、30 ％、25％、15％10

	1	5％		(5:1) or (3:1)	% or 5%
						IFN	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%%	1FN	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10％	IFN	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						IFN	Glucosan and sucrose (9:1) or 6:1	100％、80％、40 ％、20％、10％	IFN	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
IFN	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10％	IFN	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%

(2) dichloromethane, acetonitrile, ethyl acetate or their the mixed solution weight ratio that is loaded with interferon (IFN) drug particles and PLGA is 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed even suspension in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			INF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The IFN microsphere of this method preparation can be used for the frequent drug administration by injection of needs, long-term treatment antiviral such as hepatitis; Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the IFN medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment four: the preparation of erythropoietin (EPO) drug particles PLA/PLGA shell-nuclear microsphere

(1) EPO and pharmaceutic adjuvant are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						EPO	Glucosan	80%, 50%, 25 %, 10% or 5%	EPO	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1)	100%, 80%, 50 %, 25% or 10%

				Or (3:1)
						EPO	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	EPO	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50 %, 25% or 10%
EPO	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	EPO	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10%	EPO	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	EPO	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	EPO	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	EPO	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10%	EPO	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10%	EPO	Poly(ethylene oxide)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10%	EPO	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	EPO	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	EPO	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15%10 % or 5%	EPO	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10%	EPO	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan and lactose	80％、60％、50	EPO	Polypyrrole alkane ketone	50％、40％、30

	(8:1) or 6:1	%, 25% or 10%			%, 25%, 15%10 % or 5%
						EPO	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	EPO	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	EPO	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	EPO	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	EPO	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Chitosan and sucrose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	EPO	Glucosan and sorbitol (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%%	EPO	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10％	EPO	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
EPO	Glucosan and sucrose (9:1) or 6:1	100％、80％、40 ％、20％、10％	EPO	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						EPO	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10％	EPO	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%

(2) dichloromethane of erythropoietin (EP0) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/0) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles

The weight ratio of PLGA/PLA

Drug particles accounts for the percentage by weight of microsphere

EPO

1,1/2,1/3,1/4,1/5,2,3,4 or 5

20%, 60%, 80%, 70% or 99.99%

The EPO microsphere of this method preparation can be used for the renal failure basin blood that basin blood that the frequent drug administration by injection of needs, long-term treatment a variety of causes cause such as oncotherapy cause; Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the EPO medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment five: the preparation of granulocyte colony-stimulating factor G-CSF drug particles PLA/PLGA shell-nuclear microsphere

(1) granulocyte colony-stimulating factor G-CSF and pharmaceutic adjuvant are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						G-CSF	Glucosan	80%, 50%, 25 %, 10% or 5%	G-CSF	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50 %, 25% or 10 %
G-CSF	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	G-CSF	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50 %, 25% or 10 %
						G-CSF	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	G-CSF	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	G-CSF	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Poly(ethylene oxide)	50％、40％、30 ％、25％、15

					%10% or 5%
						G-CSF	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10%	G-CSF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	G-CSF	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	G-CSF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15%10 % or 5%	G-CSF	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10%	G-CSF	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50 %, 25% or 10%	G-CSF	Polypyrrole alkane ketone	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	G-CSF	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	G-CSF	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	G-CSF	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	G-CSF	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Chitosan and sucrose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	G-CSF	Glucosan and sorbitol (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%%	G-CSF	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						G-CSF	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10％	G-CSF	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
G-CSF	Glucosan and sugarcane	100％、80％、40	G-CSF	Glucosan and lysine (9:	50％、40％、30

	Sugar (9:1) or 6:1	％、20％、10％		1), (7:1), (6:1), (5:1) or (3:1)	%, 25%, 15 %10% or 5%
						G-CSF	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10％	G-CSF	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%

(2) dichloromethane of granulocyte colony-stimulating factor (G-CSF) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense is such as following:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			G-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The G-CSF microsphere of this method preparation can be used for the leukopenia that the frequent drug administration by injection of needs, long-term treatment a variety of causes cause and demonstrate,prove; Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the G-CSF medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment six: the preparation of granulocyte-macrophage colony stimutaing factor (GM-CSF) drug particles PLA/PLGA shell-nuclear microsphere

(1) granulocyte-macrophage colony stimutaing factor (GM-CSF) and pharmaceutic adjuvant are prepared into drug particles; Concrete proportioning is:

Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight
						GM-CSF	Glucosan	80%, 50%, 25%, 10% or 5%	GM-CSF	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50 %, 25% or 10%
GM-CSF	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25%, 10% or 5%	GM-CSF	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50 %, 25% or 10%

GM-CSF	Glucosan and trehalose (9:1)	80%, 50%, 25%, 10% or 5%	GM-CSF	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25% or 10%	GM-CSF	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Poly(ethylene oxide)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50%, 25% or 10%	GM-CSF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Glucosan and trehalose (8:1)	50%, 40%, 30%, 25%, 15%10% or 5 %	GM-CSF	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan and trehalose (6:1)	50%, 40%, 30%, 25%, 15%10% or 5 %	GM-CSF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30%, 25%, 15%10% or 5 %	GM-CSF	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50%, 25% or 10%	GM-CSF	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%

GM-CSF	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50%, 25% or 10%	GM-CSF	Polypyrrole alkane ketone	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25%, 10% or 5%	GM-CSF	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25%, 10% or 5%	GM-CSF	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25%, 10% or 5%	GM-CSF	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40%, 20%, 10% or 5%	GM-CSF	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Chitosan and sucrose (9:1) or 6:1	60%, 50%, 40%, 20%, 10% or 5%	GM-CSF	Glucosan and sorbitol (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Starch and lactose (9:1) or 6:1	60%, 50%, 40%, 20%, 10% or 5%%	GM-CSF	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Cellulose and lactose (9:1) or 6:1	100％、80％、 40％、20％、 10％	GM-CSF	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
GM-CSF	Glucosan and sucrose (9:1) or 6:1	100％、80％、 40％、20％、 10％	GM-CSF	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%
						GM-CSF	Starch and trehalose (9:1) or 6:1	100％、80％、 40％、20％、 10％	GM-CSF	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%

(2) dichloromethane of granulocyte-macrophage colony stimutaing factor (GM-CSF) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			GM-CSF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The GM-CSF microsphere of this method preparation goes for cancer chemotherapy and caused leukopenia with the bone marrow depression therapy time, also be applicable to treatment marrow failure patient's low leukocyte counts, the potential infection complication of possibility in the time of also can preventing leukopenia can also make the recovery of infecting the neutrophilic granulocyte minimizing that causes accelerate.Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the GM-CSF medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment seven: the preparation of recombinant human somatropin (GH) drug particles PLA/PLGA shell-nuclear microsphere

(1) recombinant human somatropin (GH) and pharmaceutic adjuvant are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						GH	Glucosan	80%, 50%, 25 %, 10% or 5%	GH	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50%, 25 % or 10%
GH	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	GH	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50%, 25 % or 10%
						GH	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	GH	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10 %	GH	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	GH	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10	GH	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%

		％
						GH	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10 %	GH	Glucosan and histidine (8:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10 %	GH	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10 %	GH	Poly(ethylene oxide)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10 %	GH	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%	GH	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%	GH	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15 %10% or 5%	GH	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10 %	GH	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50 %, 25% or 10 %	GH	Polypyrrole alkane ketone	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	GH	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	GH	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	GH	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10 % or 5%	GH	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Chitosan and sugarcane	60％、50％、40	GH	Glucosan and sorbitol (9:	50％、40％、30％、25

	Sugar (9:1) or 6:1	%, 20%, 10 % or 5%		1), (7:1), (6:1), (5:1) or (3:1)	%, 15%10% or 5%
						GH	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10 % or 5%%	GH	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	GH	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
						GH	Glucosan and sucrose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	GH	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%
GH	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	GH	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25 %, 15%10% or 5%

(2) dichloromethane of recombinant human somatropin (GH) polysaccharide microgranule and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			GH	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The GH microsphere of this method preparation goes for being used for child, adult's growth hormone deficiency, Turner's syndrome, the retardation of growth that child's chronic renal insufficiency causes, hypermetabolism state (negative nitrogen balance) after operation, the wound, burn, pyaemia septica.Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the GH medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment eight: the preparation of vascular endothelial cell growth factor (VEGF) drug particles PLA/PLGA shell-nuclear microsphere

(1) vascular endothelial cell growth factor (VEGF) and polysaccharide are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for the percentage by weight of granule gross weight
						VEGF	Glucosan	80%, 50%, 25%, 10% or 5%	VEGF	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50%, 25% or 10%
VEGF	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25%, 10% or 5%	VEGF	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50%, 25% or 10%
						VEGF	Glucosan and trehalose (9:1)	80%, 50%, 25%, 10% or 5%	VEGF	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50%, 25% or 10%	VEGF	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25% or 10%	VEGF	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50%, 25% or 10%	VEGF	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50%, 25% or 10%	VEGF	Glucosan and histidine (8:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50%, 25% or 10%	VEGF	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50%, 25% or 10%	VEGF	Poly(ethylene oxide)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50%, 25% or 10%	VEGF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Glucosan and trehalose (8:1)	50%, 40%, 30%, 25%, 15%10% or 5 %	VEGF	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan and trehalose (6:1)	50%, 40%, 30%, 25%, 15%10% or 5 %	VEGF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %

VEGF	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30%, 25%, 15%10% or 5 %	VEGF	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50%, 25% or 10%	VEGF	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50%, 25% or 10%	VEGF	Polypyrrole alkane ketone	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25%, 10% or 5%	VEGF	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25%, 10% or 5%	VEGF	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25%, 10% or 5%	VEGF	Glucosan, trehalose and sodium alginate (9:1; 1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40%, 20%, 10% or 5%	VEGF	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Chitosan and sucrose (9:1) or 6: 1	60%, 50%, 40%, 20%, 10% or 5%	VEGF	Glucosan and sorbitol (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Starch and lactose (9:1) or 6:1	60%, 50%, 40%, 20%, 10% or 5%%	VEGF	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Cellulose and lactose (9:1) or 6:1	100％、80％、 40％、20％、 10％	VEGF	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
VEGF	Glucosan and sucrose (9:1) or 6:1	100％、80％、 40％、20％、 10％	VEGF	Glucosan and lysine (9:1), (7L1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						VEGF	Starch and trehalose (9:1) or 6:1	100％、80％、 40％、20％、 10％	VEGF	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %

(2) dichloromethane of vascular endothelial cell growth factor (VEGF) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense are such as following table:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			VEGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The VEGF microsphere of this method preparation goes for the short vascular endothelial cell growth of formation treatment with new vessels.Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the GM-CSF medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment nine: the preparation of bone morphogenetic protein(BMP) (BMP) drug particles PLA/PLGA shell-nuclear microsphere

(1) bone morphogenetic protein(BMP) (BMP) and polysaccharide are prepared into drug particles; Concrete proportioning is:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						BMP	Glucosan	80%, 50%, 25 %, 10% or 5%	BMP	Cyclodextrin and sucrose (9:1), (7:1), (6:1), (5:1) or (3:1)	100%, 80%, 50%, 25% or 10%
BMP	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	BMP	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50%, 25% or 10%
						BMP	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	BMP	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10 %	BMP	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	BMP	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10	BMP	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5

		％			％
						BMP	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10 %	BMP	Glucosan and histidine (8:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10 %	BMP	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10 %	BMP	Poly(ethylene oxide)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10 %	BMP	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%	BMP	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%	BMP	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15 %10% or 5%	BMP	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10 %	BMP	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50 %, 25% or 10 %	BMP	Polypyrrole alkane ketone	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BMP	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BMP	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BMP	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10 % or 5%	BMP	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Chitosan and sugarcane	60％、50％、40	BMP	Glucosan and sorbitol (9:	50％、40％、30％、

	Sugar (9:1) or 6:1	%, 20%, 10 % or 5%		1), (7:1), (6:1), (5:1) or (3:1)	25%, 15%10% or 5 %
						BMP	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10 % or 5%%	BMP	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	BMP	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
						BMP	Glucosan and sucrose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	BMP	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %
BMP	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10 ％	BMP	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30%, 25%, 15%10% or 5 %

(2) dichloromethane of bone morphogenetic protein(BMP) (BMP) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/0) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense is such as following:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			BMP	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The BMP microsphere of this method preparation goes for the treatment behind the miscellaneous fracture.Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the BMP medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Embodiment ten: the preparation of bone-derived growth factor (BDGF) polysaccharide microgranule PLA/PLGA shell-nuclear microsphere

(1) bone-derived growth factor (BDGF) and polysaccharide are prepared into drug particles; Concrete proportioning is as follows:

Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight	Medicine	Pharmaceutic adjuvant	Medicine accounts for particulate percentage by weight
						BDGF	Glucosan	80％、50％、25	BDGF	Cyclodextrin and sucrose (9:1),	100％、80％、50

		%, 10% or 5%		(7:1), (6:1), (5:1) or (3:1)	%, 25% or 10 %
						BDGF	Glucosan, zinc salt and trehalose (9:1:1)	80%, 50%, 25 %, 10% or 5%	BDGF	Polyethylene Glycol, chitosan and sorbitol (8:1:1) or (5:1:1)	100%, 80%, 50 %, 25% or 10 %
BDGF	Glucosan and trehalose (9:1)	80%, 50%, 25 %, 10% or 5%	BDGF	Glucosan, glycine and mannitol (8:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan, magnesium salt and trehalose (9:1:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Poly(ethylene oxide) and sucrose (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Glucosan, zinc salt and mannitol (9:0.5:1)	80%, 50%, 25 % or 10%	BDGF	Polypyrrole alkane ketone and glycine (5:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Polyethylene Glycol and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Glucosan and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Poly(ethylene oxide) and zinc salt (9:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Glucosan and histidine (8:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Polyethylene Glycol and molybdenum salt (9:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Polyethylene Glycol and lysine (9:1), (8:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Poly(ethylene oxide) and calcium salt (9:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Poly(ethylene oxide)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan, zinc salt and lactose (9:0.5:1)	100%, 80%, 50 %, 25% or 10%	BDGF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Glucosan and trehalose (8:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	BDGF	Glucosan, poly(ethylene oxide) and zinc salt (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan and trehalose (6:1)	50%, 40%, 30 %, 25%, 15%10 % or 5%	BDGF	Glucosan, Polyethylene Glycol and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Cyclodextrin and trehalose (8:1) or 6:1	50%, 40%, 30 %, 25%, 15%10 % or 5%	BDGF	Glucosan, magnesium salt and trehalose (8:1) (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan, zinc salt and lactose (8:1:1)	80%, 70%, 50 %, 25% or 10%	BDGF	Glucosan, polypyrrole alkane ketone and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%

BDGF	Glucosan and lactose (8:1) or 6:1	80%, 60%, 50 %, 25% or 10%	BDGF	Polypyrrole alkane ketone	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan and trehalose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BDGF	Glucosan, starch and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Glucosan and lactose (8:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BDGF	Glucosan, chitosan and trehalose (8:1:1), (7:1:1) or (6:1:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 25 %, 10% or 5%	BDGF	Glucosan, trehalose and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Glucosan and sodium alginate (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	BDGF	Glucosan, glycine and sodium alginate (9:1:1), (8:1:1) or 4:1:1	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Chitosan and sucrose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%	BDGF	Glucosan and sorbitol (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Starch and lactose (9:1) or 6:1	60%, 50%, 40 %, 20%, 10% or 5%%	BDGF	Glucosan and lactose (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Cellulose and lactose (9:1) or 6:1	100％、80％、40 ％、20％、10％	BDGF	Glucosan and glycine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
BDGF	Glucosan and sucrose (9:1) or 6:1	100％、80％、40 ％、20％、10％	BDGF	Glucosan and lysine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%
						BDGF	Starch and trehalose (9:1) or 6:1	100％、80％、40 ％、20％、10％	BDGF	Glucosan and histidine (9:1), (7:1), (6:1), (5:1) or (3:1)	50%, 40%, 30 %, 25%, 15 %10% or 5%

(2) dichloromethane of bone-derived growth factor (BDGF) drug particles and PLGA, acetonitrile, ethyl acetate or their mixed solution weight ratio are 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8,1:9, the stirring of 1:10 equal proportion, whirlpool or formed the even suspension that gets in ultrasonic 1-5 minutes, again the organic solution of PLA is added in the above-mentioned suspension, the even suspension that gets of restir, whirlpool or formation in ultrasonic 1-5 minutes, i.e. promptly oily solid (S/O) emulsion of wrapping of oil bag solid;

The enforcement of step (3), (4), (5) is identical with embodiment one.

The microsphere that present embodiment obtains, concrete set of dispense is such as following:

Drug particles	The weight ratio of PLGA/PLA	Drug particles accounts for the percentage by weight of microsphere
			BDGF	1,1/2,1/3,1/4,1/5,2,3,4 or 5	20%, 60%, 80%, 70% or 99.99%

The BMP microsphere of this method preparation goes for the treatment behind the miscellaneous fracture.Can reduce the frequency of injection and alleviate patient's misery.High minimum can the reaching more than 80% of the microsphere envelop rate of this method preparation, dashing forward to release very little and almost do not have not exclusively discharges, and can reach zero level release.Can make simultaneously the BDGF medicine is non-inactivation in whole process of preparation and therapeutic process maintenance high activity.

Claims (10)

1, a kind of PLA/PLGA shell-nuclear microsphere with oil-in-water-oil bag solid preparation, it is characterized in that, comprise medicine, polylactic acid and/or polylactic acid-hydroxyacetic acid or/and pharmaceutic adjuvant, their weight percent content is: medicine 0.01-50%, polylactic acid and/or polylactic acid-hydroxyacetic acid 20-99.99%, pharmaceutic adjuvant are 0-30%.

2, the PLA/PLGA shell-nuclear microsphere with oil-in-water-oil bag solid preparation according to claim 1, it is characterized in that, described medicine comprises small-molecule drug and macromolecular drug, and described small-molecule drug refers to chemicals, and macromolecular drug is meant biopharmaceutical macromolecular drug.

3, the PLA/PLGA shell-nuclear microsphere with oil-in-water-oil bag solid preparation according to claim 2 is characterized in that described macromolecular drug is finger protein macromolecular drug, vaccine, antibody, nucleic acid or liposome medicament especially, wherein:

Described protein macromolecule medicine is: erythropoietin, recombinant human granulocyte colony stimulating factor, granulocyte-macrophage colony stimutaing factor, vaccine, interferon, growth hormone, insulin, epidermal growth factor, fibroblast growth factor, transforming growth factor, insulin like growth factor, vascular endothelial cell growth factor, PDGF, endothelial cell growth factor (ECGF), nerve growth factor, bone-derived growth factor, bone morphogenetic protein(BMP), tissue polypeptide antigen, antibody, blood coagulation factor VIII, or it is a kind of in the plasma thromboplastin component genetic factor;

Described nucleic acid is: antisense nucleotide, microRNA or gene.

4, the PLA/PLGA shell-nuclear microsphere with oil-in-water-oil bag solid preparation according to claim 1, it is characterized in that, it is a kind of or combination in any of little saccharide, polyhydroxy compounds, polysaccharide compound, amino-acid compound or inorganic salts material that described pharmaceutic adjuvant refers to, wherein:

Described little saccharide is a kind of in sucrose, trehalose, glucose, maltose or the lactose;

Described polyhydroxy compounds is a kind of in mannitol, sorbitol, glycerol, 1,2-propylene glycol, erythritol, Polyethylene Glycol, polyvinyl alcohol, poly(ethylene oxide) or the polypyrrole alkane ketone;

Described polysaccharide compound is a kind of in glucosan, sodium alginate, chitosan, starch, cellulose or the cyclodextrin material;

Described amino-acid compound is a kind of in glycine, lysine, arginine, glutamic acid or the histidine;

Described inorganic salts material is a kind of in zinc salt, calcium salt, mantoquita, magnesium salt or the molybdenum salt.

5, a kind of preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares as claimed in claim 1 is characterized in that may further comprise the steps:

1. medicine or medicine and pharmaceutic adjuvant are prepared into drug particles;

2. drug particles is joined the organic solution of PLA and PLGA or the organic solution of hydrophilic drug particles and PLGA mixed after, mixed with the organic solution of PLA again; Or it is mixed earlier hydrophobic drug particles and PLA, mixed with the organic solution of PLGA again; Form suspension, i.e. oil phase O₁Middle stirring or whirlpool make it homodisperse and form uniform suspension;

3. 2. completing steps is formed the aqueous solution emulsifying that microsphere suspension is added drop-wise to sodium chloride solution and surfactant, form emulsion;

4. then step emulsion 3. being transferred to sodium chloride solution solidifies;

5. completing steps sample lyophilizing is 4. obtained PLA/PLGA shell-nuclear microsphere.

6, the preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares according to claim 5, it is characterized in that, described drug particles, the size of its drug particles is at 0.2-10 μ mm, and its weight accounts for the 0.2-50% of drug particles gross weight.

7, the preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares according to claim 5, it is characterized in that the organic solution of described PLA and/or PLGA is: the organic solution of PLA and PLGA is added 0.1-20%w/w Polyethylene Glycol or poloxamer; The dichloromethane of PLA and PLGA, ethyl acetate, acetonitrile, heptane, chloroform or acetone organic solution.

8, the preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares according to claim 5, it is characterized in that the organic solution concentration of described PLA and/or PLGA is: the weight percent concentration that accounts for total oil phase of PLA is 0.5-80%; The weight percent concentration that PLGA accounts for total oil phase is 20-99.5%.

9, the preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares according to claim 5, it is characterized in that, described surfactant is polyvinyl alcohol, Polyethylene Glycol, polyvinylpyrrolidone, poloxamer, poly-sorbic alcohol, ethyl cellulose or tween, and wherein: the concentration of surface active agent polyvinyl alcohol is 0.5-10% or contains 0.5-10% sodium chloride salt solution; The concentration of Polyethylene Glycol is 0.5-20% or contains the saline solution of 0.5-10% sodium chloride; Polyvinylpyrrolidone concentration is 0.5-20% or contains 0.5-10% sodium chloride salt solution; Poloxamer concentration is 0.5-20% or contains 0.5-10% sodium chloride salt solution; Poly-sorbic alcohol concentration is 0.5-20% or contains 0.5-10% sodium chloride salt solution; Ethyl cellulose concentration is 0.5-20% or contains 0.5-10% sodium chloride salt solution that above concentration is weight percent concentration.

10, the preparation method with oil-in-water-PLA/PLGA shell-nuclear microsphere that oil bag solid prepares according to claim 5 is characterized in that the particle diameter of described microsphere is 1-500 μ m.