CN101721375A - Insulin slow release micron sphere composition and preparation method thereof - Google Patents
Insulin slow release micron sphere composition and preparation method thereof Download PDFInfo
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- CN101721375A CN101721375A CN200910312647A CN200910312647A CN101721375A CN 101721375 A CN101721375 A CN 101721375A CN 200910312647 A CN200910312647 A CN 200910312647A CN 200910312647 A CN200910312647 A CN 200910312647A CN 101721375 A CN101721375 A CN 101721375A
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Abstract
The invention provides an insulin slow release micron sphere composition and a preparation method thereof, belonging to the field of biotechnology pharmacy. The composition has the following components by weight percent: 50-99.8% of polylactic acid-glycollic acid, polylactic acid or polycaprolactone, and 0.2-50% of insulin nano sphere composition. The preparation method comprises two steps: 1) preparation of insulin nano sphere composition; and 2) preparation of insulin slow release micron sphere composition. The invention overcomes the defects that spherical micro nanospheres can not be prepared by the prior art, the stability of the micro nanospheres in high polymer slow and controlled release material can not be realized, the entrapment rate is low, S/O/O is in serious burst, and environment pollution is caused. The micron sphere prepared by the method in the invention has controllable particle size, no environment pollution, smooth and rounding surface and structured particles without adhesion, the particle size can be regulated and controlled from 1mu m to 200 mu m as required, and the freeze-drying powder is white, fine and smooth, loose, is without collapse and adhesion, and has good redispersibility.
Description
Technical field
What the present invention relates to is a kind of composition and method of making the same of biological pharmacy technical field, especially a kind of insulin slow release micron sphere composition and preparation method thereof.
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 slow release or controlled release form; For some tumor treatment, need some drug targetings in the disease photograph, for example targeting is in tumor vascular thromboembolism micron ball 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 micron ball of active loss such as W/O/W method preparation in preparation microgranule process.It is imperative that the albumen micron ball 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 micron of preparation ball in order to reduce prominent releasing, but still can not solve this difficult problem up till now.
Find by prior art documents, [Claudia Klingler, Bernd W.M ü ller, HartwigSteckel, Insulin-micro-and nanoparticles for pulmonary delivery, InternationalJournal of Pharmaceutics, 377 (2009) 173-179], [Claudia Klingler, BerndW.M ü ller, Hartwig Steckel, the micro-nano grain of rice of insulin is used for pulmonary administration, the Inpharm magazine, 377 (2009) 173-179] people such as Claudia Klingler reported that in the document granule that utilizes spray-dired method to prepare 2 μ m is used for pulmonary administration, but at drug safety, conveniently still have very large problem, say as the medical science professor Mayer Davidson of Los Angeles charles De Lu university (Charles R.DrewUniversity) that had once participated in the Exubera early studies in man: " volume ratio of Exubera (inhalant of insulin) is bigger, uses inconvenience.In addition, use the patient of Exubera also to need to use protamine zine insulin to keep the level of blood glucose, and need regularly carry out lung function." Exubera of Pfizer removes after the city, Novo Nordisk has also stopped the exploitation of imbedibility insulin.The multi-emulsion method that the document utilizes W/O/W to see most prepares double-deck micron ball, and the oil-water interfaces of multi-emulsion method are the albumen killers who generally acknowledges, cause the gathering of water miscible albumen at this interface, cause envelop rate also not high equally, exist not exclusively to discharge and prominent releasing.[Yadong Hah, Huayu Tian, Pan He, Xuesi Chen, Xiabin Jing, Insulin nanoparticle preparation and encapsulation intopoly (lactic-co-glycolic acid) microspheres by using an anhydrous system, International Journal of Pharmaceutics 378 (2009) 159-166], [people such as Yadong Hah has reported the nanoparticle of preparation insulin and has utilized the S/O/O emulsion process further to be wrapped in the PLGA micron ball in the document, the Inpharm magazine, 378 (2009) 159-166].They utilize document [Zhang, X.F., Hu, J.L., Chen, X.S., Jing, X.B, 2005.Preparation and characteri-zation of biodegradableinsulin-loaded microspheres.Chem.J.Chin.U 26,554? 57.] the method isoelectric point precipitation prepare nanoparticle, and then the microcapsule bag forms PLGA micron ball, but the granule of the nanometer insulin of preparation still is not spherical, but cube shaped and other irregular shape, this also is unfavorable for further sealing; Preparation micron ball has utilized a large amount of organic solvent N that is difficult to remove, and these solvents such as dinethylformamide (DMF), Semen Maydis oil, ether and ethanol cause shortcomings such as great quality control trouble to the quality of environment and micron ball.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of insulin slow release micron sphere composition and preparation method thereof is provided.The invention solves existing technology and can't prepare the difficult problem of spheric nanometer insulin for a long time, utilize oil-in-water-oil bag solid (S/O/W) preparation micron ball method that the further microcapsule of above-mentioned nanosphere is wrapped in the macromolecular material with slow release.Make the micron ball smooth surface rounding 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:
The insulin slow release micron sphere composition that the present invention relates to, its component and percentage by weight are: polylactic acid-glycolic guanidine-acetic acid (PLGA), polylactic acid (PLA) or polycaprolactone (PCL) are that 50%-99.8%, insulin nanosphere compositions are 0.2%-50%.
Described insulin nanosphere compositions, its component and percentage by weight are: insulin is that 50%-99.99%, glucosan are that 0%-50%, zinc ion chemical compound are 0.01%-50%.
The particle diameter of described insulin slow release micron sphere composition is 1-500 μ m.
Described zinc ion chemical compound is zinc acetate, zinc chloride, zinc sulfate, hydrogen sulfate zinc, zinc nitrate or zinc gluconate.
The preparation method of the aforesaid insulin slow release micron sphere composition that the invention still further relates to may further comprise the steps:
1. insulin nanosphere preparation of compositions
A) percentage by weight that will account for insulin nanosphere compositions in advance is that the chemical compound of 0.01%-50% zinc ion is mixed with certain concentration solution or above-mentioned zinc ion chemical compound and contains the percentage by weight that accounts for insulin nanosphere compositions is mixed with certain density glucosan and zinc ion chemical compound for the 0%-50% glucosan solution;
B) drip that to contain the percentage by weight that accounts for insulin nanosphere compositions be that the insulin of 50%-99.99% is dissolved in the PEG solution of acidity (hydrochloric acid solution, pH＜2) and then in above-mentioned steps solution a);
C) and then with above-mentioned steps b) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
D) with above-mentioned steps c) powder after the lyophilizing utilizes organic solvent dissolution Polyethylene Glycol (PEG), centrifugally again remove the organic solution that supernatant is PEG, triplicate, the dried collection of volatilization is removed poly-ethanol (PEG) with organic solvent then and is promptly got insulin nanosphere compositions then;
2. insulin slow release micron sphere composition preparation
A) percentage by weight that 1. step must be accounted for the islets of langerhans slow release micron sphere composition is that 2%-50% insulin nanosphere compositions joins after the percentage by weight that accounts for insulin slow release micron sphere composition is 50%-98% polylactic acid-glycolic guanidine-acetic acid (PLGA), polylactic acid (PLA) or polycaprolactone (PCL) organic solution, forms suspension.Be to stir in the oil phase (O) or whirlpool etc. makes it homodisperse and forms uniform suspension;
B) completing steps a) being formed the suspension of micron ball, to be added to sodium chloride solution and the weight percent concentration that weight percent concentration is 0%-10% be 1%-10% surfactant emulsifying 1-5min, and then it being transferred to weight percent concentration is that the sodium chloride solution of 1%-10% solidified 1-4 hour; The micron ball that to collect then washes three to five times with water and removes surfactant and sodium chloride;
C) with completing steps b) the sample lyophilizing remove moisture and get insulin slow release micron sphere composition;
The weight percent concentration of described insulin is: 0.001%20%
The chemical compound of described zinc ion is: the weight percent concentration of zinc acetate, zinc chloride, zinc sulfate, hydrogen sulfate zinc, zinc nitrate or zinc gluconate is: 0.001%-20%;
The weight percent concentration of described glucosan is: 0.001%-20%, the molecular weight of glucosan are 10000-5000000 dalton;
The molecular weight of described PEG is: 2, and 000-300,000 dalton, the weight percent concentration in aqueous solution is 1%-40%;
Described insulin nanosphere compositions, its particle grain size size are that 0.02 μ m-1 μ mm is good at 0.01 μ m-2 μ m with particle diameter.
Described oil phase (O) is: the organic solution of polylactic-co-glycolic acid (PLGA), polylactic acid (PLA), polycaprolactone (PCL) or their any mixture;
Organic solvent in the described organic solution is: dichloromethane, ethyl acetate, acetonitrile, heptane, chloroform or acetone organic solution are good with the organic solution of dichloromethane, ethyl acetate, acetonitrile or their combination in any;
The organic solution concentration of described polylactic-co-glycolic acid (PLGA), polylactic acid (PLA), polycaprolactone (PCL) or their any mixture is: the weight percent concentration of polylactic-co-glycolic acid (PLGA), polylactic acid (PLA), polycaprolactone (PCL) or their any mixture is 5%40%;
Described polylactic-co-glycolic acid (PLGA), polylactic acid (PLA), their molecular weight of polycaprolactone (PCL) are 5000-500000 dalton;
Described surfactant is: polyvinyl alcohol (PVA), Polyethylene Glycol (PEG), polyvinylpyrrolidone (PVP) or poloxamer (poloxmer);
Described polyvinyl alcohol (PVA), Polyethylene Glycol (PEG), polyvinylpyrrolidone (PVP) or their molecular weight of poloxamer (poloxmer) are respectively: 10000-1000000 dalton, 4000-300000 dalton, 20000-400000 dalton or 10000-400000 dalton;
The weight percent concentration of described surface active agent polyvinyl alcohol (PVA) is 0.5%-10% or the solution that contains 0.5%-10% sodium chloride, the weight percent concentration of Polyethylene Glycol (PEG) is 0.5%-20% or the saline solution that contains 0.5%-10% sodium chloride, polyvinylpyrrolidone (PVP) weight percent concentration is 0.5%-20% or to contain weight percent concentration be saline solution such as 0.5%-10% sodium chloride, poloxamer (poloxmer) weight percent concentration is 0.5%-20% or contains saline solution such as 0.5%-10% sodium chloride.
The present invention overcomes the nanosphere that existing technology does not prepare spheric insulin, can be prepared into fairly regular micro-nano ball compositions, can overcome the unstability of insulin in slow controlled-release material in addition; Be prepared into the micro-nano ball compositions, avoid not high, and the serious prominent of S/O/O released the shortcoming of the environmental pollution that causes with the envelop rate of conventional W/O and W/O/W; Adopt this method to prepare micron sphere composition, 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 insulin, especially those physicochemical properties are unsettled.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.
Description of drawings
Fig. 1 insulin nanosphere compositions sem photograph.
Fig. 2 insulin slow release PLGA micron sphere composition sem photograph.
Fig. 3 insulin slow release PLGA micron sphere composition release in vitro curve chart.
The specific embodiment
Below embodiments of the invention are elaborated: following examples have provided detailed embodiment and process being to implement under the prerequisite with the technical solution of the present invention, but protection scope of the present invention is not limited to following embodiment.
Embodiment one:
1. insulin nanosphere preparation of compositions
A) in advance zinc acetate is dissolved in water, be mixed with weight percent concentration and be respectively solution such as 0.001%, 0.001%, 10% or 20%;
B) insulin being dissolved in hydrochloric acid solution (pH＜2) weight percent concentration is respectively: 1%, 10%, 20% or 40% PEG (molecular weight is 8000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 3 or 1: 4 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution PEG, centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.99%, 75% or 80% then; Zinc acetate is 50%, 0.01%, 25% or 20% accordingly; Glucosan is zero entirely); (wherein according to the insulin nanosphere of preparation in 1: 10 as shown in Figure 1, particulate smooth surface, particle size distribution are even, about 0.01-0.03 μ m; Other 1: 1,1: 3 or the particulate surface of insulin nanosphere of preparation in 1: 4 also more even to 1: 10 similar smoother, particle size distribution, particle diameter is about 0.3-1 μ m, 0.5-1.5 μ m and 0.4-2 μ m respectively, all demonstrations on the figure);
2. insulin slow release micron sphere composition preparation
A) will 1. must account for insulin nanosphere compositions and polylactic acid (PLA) weight percent concentration is that 5%, 10%, 20% or 40% dichloromethane, acetonitrile or ethyl acetate solution weight ratio are that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 75% or 98%), whirlpool or ultrasonic 1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to Polyethylene Glycol (PVA) aqueous solution of 1%-10% sodium chloride and 0%-10% and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that the 1000ml sodium chloride solution of 1%-10% solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and washing with water 3-5 time, (the micron ball of ratio preparation in 1: 20 as shown in Figure 2, smooth surface, the particle size distribution of micron ball are even, and particle diameter is about 40-100 μ m to obtain micron sphere composition after the lyophilizing; The particle diameter of the micron ball of other corresponding proportion 3: 20,1: 15 or preparation in 2: 245 is about 60-150 μ m, 100-200 μ m and 200-500 μ m respectively, does not all show on the figure).
The micron ball of 1: 20 ratio preparation at the release in vitro curve as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure.
Embodiment two:
1. insulin nanosphere preparation of compositions
A) in advance zinc sulfate being dissolved in weight concentration respectively is that 0%, 0.001%, 5%, 10% or 20% molecular weight is in 10000 the dextran solution, is mixed with weight percent concentration and is respectively solution such as 0.001%, 0.001%, 5%, 10% or 20%;
B) insulin being dissolved in acidity (hydrochloric acid solution, pH＜2) weight percent concentration is respectively: 1%, 5%, 10%, 20% or 40% PEG (molecular weight is 6000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 5%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 8,1: 3 or 3: 14 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution PEG, centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.98%, 80%, 60% or 70% then; Glucosan is respectively: 0%, 0.01%, 10%, 20% or 15%; Zinc sulfate is respectively: 50%, 0.01%, 10%, 20% or 15%); (result is as shown in Figure 1 similar, particulate smooth surface, particle size distribution are even, are about 0.01-0.04 μ m, 0.02-0.06 μ m, 0.3-1 μ m, 0.5-1.5 μ m and 0.4-2 μ m respectively according to the particle diameter of the insulin nanosphere of 1: 1,1: 10,1: 8,1: 3 or preparation in 3: 14.)
2. insulin slow release micron sphere composition preparation
A) dichloromethane, acetonitrile or the ethyl acetate solution weight ratio that will 1. must account for insulin nanosphere compositions and polylactic acid-glycolic guanidine-acetic acid (PLGA) weight percent concentration 5%, 10%, 15%, 20% or 40% is that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 85%, 75% or 98%), whirlpool or ultrasonic 0.1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 9,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to Polyethylene Glycol (PVA) solution of 1%-10% sodium chloride and 1%-10% and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) do you the emulsion of step (b) is added to the 100ml that concentration is 1%-10%? the 000ml sodium chloride solution solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and wash with water 3-5 time, (micron of aforementioned proportion preparation is ball seemingly as shown in Figure 2 to obtain micron sphere composition after the lyophilizing, smooth surface, the particle size distribution of micron ball are even, the about respectively 1-10 μ of particle diameter m, 10-50 μ m, 80-120 μ m, 100-200 μ m and the 200-500 μ m of the micron ball of corresponding proportion 1: 20,3: 20,1: 9,1: 15 or preparation in 2: 245 all do not show on the figure).
Like as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure to the micron ball of corresponding proportion preparation at the release in vitro class of a curve.
Embodiment three:
1. insulin nanosphere preparation of compositions
A) in advance zinc nitrate being dissolved in weight concentration respectively is that 0%, 0.001%, 5%, 10% or 20% molecular weight is in 100000 the dextran solution, is mixed with weight percent concentration and is respectively solution such as 0.001%, 0.001%, 5%, 10% or 20%;
B) insulin being dissolved in acidity (hydrochloric acid solution, pH＜2) weight percent concentration is respectively: 1%, 5%, 10%, 20% or 40% PEG (molecular weight is 4000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 5%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 8,1: 3 or 3: 14 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution PEG, centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.98%, 80%, 60% or 70% then; Glucosan is respectively: 0%, 0.01%, 10%, 20% or 15%; Zinc nitrate is respectively: 50%, 0.01%, 10%, 20% or 15%); (result is as shown in Figure 1 similar, particulate smooth surface, particle size distribution are even, are about 0.015-0.04 μ m, 0.03-0.06 μ m, 0.35-1 μ m, 0.52-1.5 μ m and 0.45-2 μ m respectively according to the particle diameter of the insulin nanosphere of 1: 1,1: 10,1: 8,1: 3 or preparation in 3: 14.)
2. insulin slow release micron sphere composition preparation
A) will 1. must account for insulin nanosphere compositions and polylactic acid-glycolic guanidine-acetic acid (PLGA) weight percent concentration is that 5%, 10%, 15%, 20% or 40% dichloromethane, acetonitrile or ethyl acetate solution weight ratio are that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 85%, 75% or 98%), whirlpool or ultrasonic 1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 9,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to Polyethylene Glycol (PEG) solution of 1%-10% sodium chloride and 1%-10% and formed emulsion in stirring, whirlpool or ultrasonic 0.1-5 minute;
(c) emulsion of step (b) being added to concentration is that the 1000ml sodium chloride solution of 1%-10% solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and wash with water 3-5 time, micron sphere composition obtained after the lyophilizing.(micron of aforementioned proportion preparation is ball seemingly as shown in Figure 2, smooth surface, the particle size distribution of micron ball are even, the about respectively 1-10 μ of particle diameter m, 10-50 μ m, 80-120 μ m, 100-200 μ m and the 200-500 μ m of the micron ball of corresponding proportion 1: 20,3: 20,1: 9,1: 15 or preparation in 2: 245 all do not show on the figure).
Like as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure to the micron ball of corresponding proportion preparation at the release in vitro class of a curve.
Embodiment four:
1. insulin nanosphere preparation of compositions
A) in advance hydrogen sulfate zinc being dissolved in weight concentration respectively is that 0%, 0.001%, 5%, 10% or 20% molecular weight is in the dextran solution of 60000-70000, is mixed with weight percent concentration and is respectively solution such as 0.001%, 0.001%, 5%, 10% or 20%;
B) insulin being dissolved in acidity (hydrochloric acid solution, pH＜2) weight percent concentration is respectively: 1%, 5%, 10%, 20% or 40% PEG (molecular weight is 2000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 5%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 8,1: 3 or 3: 14 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution PEG, centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.98%, 80%, 60% or 70% then; Glucosan is respectively: 0%, 0.01%, 10%, 20% or 15%; Zinc nitrate is respectively: 50%, 0.01%, 10%, 20% or 15%); (result is as shown in Figure 1 similar, particulate smooth surface, particle size distribution are even, are about 0.010-0.035 μ m, 0.025-0.06 μ m, 0.34-1 μ m, 0.50-1.5 μ m and 0.45-2 μ m respectively according to the particle diameter of the insulin nanosphere of 1: 1,1: 10,1: 8,1: 3 or preparation in 3: 14.)
2. insulin slow release micron sphere composition preparation
A) will 1. must account for insulin nanosphere compositions and poly-own lactone (PCL) weight percent concentration is that 5%, 10%, 15%, 20% or 40% dichloromethane, acetonitrile or ethyl acetate solution weight ratio are that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 85%, 75% or 98%), whirlpool or ultrasonic 1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 9,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to polyvinylpyrrolidone (PVP) solution of 1%-10% sodium chloride and 1%-10% and formed emulsion in stirring, whirlpool or ultrasonic 1-5 minute;
(c) do you the emulsion of step (b) is added to the 100ml that concentration is 1%-10%? the 000ml sodium chloride solution solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and wash with water 3-5 time, (micron of aforementioned proportion preparation is ball seemingly as shown in Figure 2 to obtain micron sphere composition after the lyophilizing, smooth surface, the particle size distribution of micron ball are even, the about respectively 40-100 μ of particle diameter m, 60-150 μ m, 80-120 μ m, 100-200 μ m and the 200-500 μ m of the micron ball of corresponding proportion 1: 20,3: 20,1: 9,1: 15 or preparation in 2: 245 all do not show on the figure).
Like as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure to the micron ball of corresponding proportion preparation at the release in vitro class of a curve.
Embodiment five:
1. insulin nanosphere preparation of compositions
A) in advance zinc gluconate being dissolved in weight concentration respectively is that 0%, 0.001%, 5%, 10% or 20% molecular weight is in 500000 the dextran solution, is mixed with weight percent concentration and is respectively solution such as 0.001%, 0.001%, 5%, 10% or 20%;
B) insulin being dissolved in acidity (hydrochloric acid solution, pH＜2) weight percent concentration is respectively: 1%, 5%, 10%, 20% or 40% PEG (molecular weight is 300000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 5%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 8,1: 3 or 3: 14 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution PEG, centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.98%, 60% or 80% then; Glucosan is respectively: 0%, 0.002%, 10% or 20%); (as shown in Figure 1, particulate smooth surface, particle size distribution are even, about 0.01-0.03 μ m); (wherein particulate smooth surface, particle size distribution were even, about 0.01-0.03 μ m as shown in Figure 1 in 1: 10; Other 1: 1,1: 3 or 1: 4 particulate surface also more even to 1: 10 similar smoother, particle size distribution, particle diameter then be respectively approximately 0.3-1 μ m, 0.5-1.5 μ m and 0.4-2 μ m)
2. insulin slow release micron sphere composition preparation
A) will 1. must account for insulin nanosphere compositions and polylactic acid (PLA) weight percent concentration is that 5%, 10%, 15%, 20% or 40% dichloromethane, acetonitrile or ethyl acetate solution weight ratio are that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 85%, 75% or 98%), whirlpool or ultrasonic 0.1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 9,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to Polyethylene Glycol (PEG) solution of 1%-10% sodium chloride and 1%-10% and formed emulsion in stirring, whirlpool or ultrasonic 1-5 minute;
(c) do you the emulsion of step (b) is added to the 100ml that concentration is 1%-10%? the 000ml sodium chloride solution solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and wash with water 3-5 time, (micron of aforementioned proportion preparation is ball seemingly as shown in Figure 2 to obtain micron sphere composition after the lyophilizing, smooth surface, the particle size distribution of micron ball are even, the about respectively 1-10 μ of particle diameter m, 10-50 μ m, 50-120 μ m, 100-200 μ m and the 200-500 μ m of the micron ball of corresponding proportion 1: 20,3: 20,1: 9,1: 15 or preparation in 2: 245 all do not show on the figure).
Like as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure to the micron ball of corresponding proportion preparation at the release in vitro class of a curve.
Embodiment six:
1. insulin nanosphere preparation of compositions
A) in advance zinc acetate being dissolved in weight concentration respectively is that 0%, 0.001%, 5%, 10% or 20% molecular weight is in 5000000 the dextran solution, is mixed with weight percent concentration and is respectively solution such as 0.001%, 0.001%, 5%, 10% or 20%;
B) insulin being dissolved in acidity (hydrochloric acid solution, pH＜2) weight percent concentration is respectively: 1%, 5%, 10%, 20% or 40% PEG (molecular weight is 300000 dalton) is mixed with weight percent concentration and is respectively: 0.001%, 1%, 5%, 10% or 20% etc.;
C) and then above-mentioned steps a) and b) solution be respectively 1: 1,1: 10,1: 8,1: 3 or 3: 14 etc. according to weight ratio the solution of step a) dripped b) solution in and constantly stir;
D) and then with above-mentioned steps c) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
E) with above-mentioned steps d) powder after the lyophilizing utilizes organic solvent dissolution Polyethylene Glycol (PEG), centrifugally again remove the organic solution that supernatant is PEG, triplicate, volatilization do to be collected then and to be removed PEG with organic solvent and promptly get insulin nanosphere compositions and (promptly be prepared into the percentage by weight that insulin accounts for insulin nanosphere compositions and be respectively 50%, 99.98%, 60% or 80% then; Glucosan is respectively: 0%, 0.002%, 10% or 20%); (as shown in Figure 1, particulate smooth surface, particle size distribution are even, about 0.01-0.03 μ m); (wherein particulate smooth surface, particle size distribution were even, about 0.01-0.03 μ m as shown in Figure 1 in 1: 10; Other 1: 1,1: 3 or 1: 4 particulate surface also more even to 1: 10 similar smoother, particle size distribution, particle diameter then be respectively approximately 0.3-1 μ m, 0.5-1.5 μ m and 0.4-2 μ m)
2. insulin slow release micron sphere composition preparation
A) will 1. must account for insulin nanosphere compositions and polylactic acid-glycolic guanidine-acetic acid (PLGA) weight percent concentration is that 5%, 10%, 15%, 20% or 40% dichloromethane, acetonitrile or ethyl acetate solution weight ratio are that (percentage by weight of the shared insulin slow release micron sphere composition of corresponding insulin nanosphere compositions is: formed even suspension in ratio stirring 50%, 60%, 85%, 75% or 98%), whirlpool or ultrasonic 1-5 minute, i.e. oil bag solid (S/O) emulsion for 1: 20,3: 20,1: 9,1: 15 or 2: 245;
B) step (a) being got emulsion droplets is added to polyvinyl alcohol (PVA) solution of 1%-10% sodium chloride and 1%-10% and formed emulsion in stirring, whirlpool or ultrasonic 1-5 minute;
(c) do you the emulsion of step (b) is added to the 100ml that concentration is 1%-10%? the 000ml sodium chloride solution solidified 1-4 hour;
(d) the centrifugal collection micron ball that step (c) is obtained, and wash with water 3-5 time, (micron of aforementioned proportion preparation is ball seemingly as shown in Figure 2 to obtain micron sphere composition after the lyophilizing, smooth surface, the particle size distribution of micron ball are even, the about respectively 1-10 μ of particle diameter m, 10-50 μ m, 50-120 μ m, 100-200 μ m and the 200-500 μ m of the micron ball of corresponding proportion 1: 20,3: 20,1: 9,1: 15 or preparation in 2: 245 all do not show on the figure).
Like as shown in Figure 3, the result shows and not prominently releases and not exclusively discharge that other also has similar result but shows on the figure to the micron ball of corresponding proportion preparation at the release in vitro class of a curve.
Claims (10)
1. insulin slow release micron sphere composition, it is characterized in that, described compositions particle diameter is: 1-500 μ m, its component and percentage by weight are: polylactic acid-glycolic guanidine-acetic acid, polylactic acid or polycaprolactone are that 50%-98%, insulin nanosphere compositions are 2%-50%.
2. the described preparation insulin slow release micron sphere composition of claim 1, it is characterized in that, described insulin nanosphere compositions, its component and percentage by weight are: insulin is that 50%-99.99%, glucosan are that 0%-50%, zinc ion chemical compound are 0.01%-50%.
3. the described preparation insulin slow release micron sphere composition of claim 2 is characterized in that, described zinc ion chemical compound is zinc acetate, zinc chloride, zinc sulfate, hydrogen sulfate zinc, zinc nitrate or zinc gluconate.
4. the preparation method of an insulin slow release micron sphere composition as claimed in claim 1 is characterized in that, comprises the steps:
1. insulin nanosphere preparation of compositions
A) percentage by weight that will account for insulin nanosphere compositions in advance is that the chemical compound of 0.01%-50% zinc ion is mixed with solution or zinc ion chemical compound and contains the percentage by weight that accounts for insulin nanosphere compositions is mixed with glucosan and zinc ion chemical compound for the 0%-50% glucosan solution;
B) drip that to contain the percentage by weight that accounts for insulin nanosphere compositions be that the insulin of 50%-99.99% is dissolved in the tart PEG solution of pH＜2 hydrochloric acid solutions and then in above-mentioned steps solution a);
C) and then with above-mentioned steps b) mix uniformly refrigerating chamber pre-freeze 8-32 hour, lyophilizing then,
D) with above-mentioned steps c) powder after the lyophilizing utilizes the organic solvent dissolution Polyethylene Glycol, centrifugally again remove the organic solution that supernatant is a Polyethylene Glycol, triplicate, the dried collection of volatilization is removed Polyethylene Glycol with organic solvent then and is promptly got insulin nanosphere compositions then;
2. insulin slow release micron sphere composition preparation
A) percentage by weight that 1. step must be accounted for the islets of langerhans slow release micron sphere composition is that 2%-50% insulin nanosphere compositions joins after the percentage by weight that accounts for insulin slow release micron sphere composition is 50%-98% polylactic acid-glycolic guanidine-acetic acid, polylactic acid or polycaprolactone organic solution, forms suspension;
B) suspension that completing steps a) is formed the micron ball is added to weight percent concentration for being 1%-10% surfactant emulsifying 1-5min less than 10% sodium chloride solution and weight percent concentration, and then it being transferred to weight percent concentration is that the sodium chloride solution of 1%-10% solidified 1-4 hour; The micron ball that to collect then washes three to five times with water and removes surfactant and sodium chloride;
C) with completing steps b) the sample lyophilizing remove moisture and get insulin slow release micron sphere composition.
5. the preparation method of insulin slow release micron sphere composition according to claim 4, it is characterized in that the chemical compound of described zinc ion is: the weight percent concentration of zinc acetate, zinc chloride, zinc sulfate, hydrogen sulfate zinc, zinc nitrate, zinc gluconate or their any mixture is: 0.001%-20%.
6. the preparation method of insulin slow release micron sphere composition according to claim 4 is characterized in that, the weight percent concentration of described glucosan is: 0.001%-20%, the molecular weight of glucosan are 10000-500000.
7. the preparation method of insulin slow release micron sphere composition according to claim 4 is characterized in that, the molecular weight of described Polyethylene Glycol is: 2, and 000-300,000 dalton, the weight percent concentration in aqueous solution is 1%-40%.
8. the preparation method of insulin slow release micron sphere composition according to claim 4 is characterized in that, the organic solvent in the described organic solution is: dichloromethane, ethyl acetate, acetonitrile, heptane, chloroform or acetone.
9. the preparation method of insulin slow release micron sphere composition according to claim 4, it is characterized in that the organic solution concentration of described polylactic acid-glycolic guanidine-acetic acid, polylactic acid, polycaprolactone or their any mixture is: the weight percent concentration of polylactic acid-glycolic guanidine-acetic acid, polylactic acid, polycaprolactone or their any mixture is 5%-40%.
10. the preparation method of insulin slow release micron sphere composition according to claim 4 is characterized in that, described surfactant is: polyvinyl alcohol, Polyethylene Glycol, polyvinylpyrrolidone or poloxamer.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102614498A (en) * | 2011-01-28 | 2012-08-01 | 四川科伦药物研究有限公司 | Insulin nanoparticle and preparation method thereof |
| CN102885783A (en) * | 2012-09-26 | 2013-01-23 | 复旦大学附属金山医院 | Nanometer medicament microspheres |
| CN108778246A (en) * | 2016-01-07 | 2018-11-09 | 美药星制药股份有限公司 | The high-purity inhalable particles and its high efficiency preparation method of insulin and insulin analog |
| CN110840828A (en) * | 2018-08-20 | 2020-02-28 | 浙江长典医药有限公司 | Preparation method of zinc gluconate oral liquid |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105963258B (en) | 2016-04-26 | 2021-01-22 | 广州帝奇医药技术有限公司 | Preparation method of sustained-release particles |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102614498A (en) * | 2011-01-28 | 2012-08-01 | 四川科伦药物研究有限公司 | Insulin nanoparticle and preparation method thereof |
| CN102614498B (en) * | 2011-01-28 | 2014-12-17 | 四川科伦药物研究有限公司 | Insulin nanoparticle and preparation method thereof |
| CN102885783A (en) * | 2012-09-26 | 2013-01-23 | 复旦大学附属金山医院 | Nanometer medicament microspheres |
| CN102885783B (en) * | 2012-09-26 | 2014-04-16 | 复旦大学附属金山医院 | Nanometer medicament microspheres |
| CN108778246A (en) * | 2016-01-07 | 2018-11-09 | 美药星制药股份有限公司 | The high-purity inhalable particles and its high efficiency preparation method of insulin and insulin analog |
| CN108778246B (en) * | 2016-01-07 | 2023-04-18 | 美药星制药股份有限公司 | High-purity inhalable particles of insulin and insulin analogs and efficient preparation method thereof |
| CN110840828A (en) * | 2018-08-20 | 2020-02-28 | 浙江长典医药有限公司 | Preparation method of zinc gluconate oral liquid |
| CN110840828B (en) * | 2018-08-20 | 2021-12-21 | 浙江长典药物技术开发有限公司 | Preparation method of zinc gluconate oral liquid |
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