CN106075400B - BMP protein preparation and preparation method thereof - Google Patents
BMP protein preparation and preparation method thereof Download PDFInfo
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- CN106075400B CN106075400B CN201610669266.4A CN201610669266A CN106075400B CN 106075400 B CN106075400 B CN 106075400B CN 201610669266 A CN201610669266 A CN 201610669266A CN 106075400 B CN106075400 B CN 106075400B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1875—Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
Abstract
The invention discloses a BMP protein preparation and a preparation method thereof. The formulation includes BMP protein, excipients, phospholipids and polysaccharides. The preparation method is that the BMP protein, the excipient, the phospholipid and the polysaccharide are stirred evenly by a glass stirring rod under the condition of 0 ℃. The BMP protein preparation of the invention can be exposed to normal temperature air for more than 10 days in the process of orthopedic surgery treatment, ensures smooth operation, can be stored for more than 30 days under low temperature condition, can be stored for a plurality of years under freezing condition, and has no change in activity.
Description
Technical Field
The invention relates to a protein stabilizer, in particular to a bone morphogenetic protein BMP preparation and a preparation method thereof.
Background
Bone Morphogenetic Proteins (BMPs) are capable of inducing the differentiation of mesenchymal cells of animals or humans into bone, cartilage, ligaments, tendons and neural tissues. BMPs are of unlimited clinical value. The safety and efficacy of genetically recombinant BMP-2 and BMP-7 has been well established in recent years, and the U.S. FDA also approved recombinant bone morphogenetic protein-2 for spinal fusion in 7 months 2002. The safety and high efficiency of inducing osteogenic activity based on BMP are confirmed by more and more experiments, and bone morphogenetic proteins have entered large-scale clinical experimental stages in some countries.
As an effective protein for orthopedic treatment, the stability of the protein needs to be ensured. In treatment, in order to achieve the effect, the bone protein needs to be exposed in vitro for a period of several days, the bone protein is delivered through a polymer drug and released in vivo to play a role, the BMP protein stored in an organic solvent can keep activity for a certain time, and freeze-dried powder storage also has a certain effect. Patent 200680021260.0 discloses a bone morphogenetic protein, which is lyophilized, with lyoprotectant and oxidation/reduction stabilizer added. The freeze-drying method is beneficial to long-term storage, but is not beneficial to short-term room-temperature storage, and the damage to protein is irreversible in the freeze-thawing process, so that the treatment effect is greatly reduced.
Disclosure of Invention
The present invention is directed to overcoming the above-mentioned drawbacks of the prior art and solving the problem of rapid decrease in activity of BMP proteins caused by short-term storage at room temperature and long-term exposure to the outside of the body during surgery by providing a BMP protein preparation.
A BMP protein preparation comprises BMP protein, excipient, phospholipid and polysaccharide.
The excipient is one or more selected from glycerol, gelatin, maltodextrin, sodium starch glycolate, hydroxypropyl methyl cellulose, sucrose stearate, starch octenyl succinate and mPEG-PLA-lysine.
The polysaccharide is one or more than one of trehalose, undaria pinnatifida polysaccharide, isomaltose hypgather, arabinogalactan and beta-1, 6-glucan.
The phospholipid is selected from one or more of lecithin, cephalin, cardiolipin, sphingomyelin, phosphatidylserine and phosphatidylglycerol.
The excipient accounts for 30-60% of the total weight of the BMP protein preparation.
The phospholipid accounts for 10-20% of the total weight of the BMP protein preparation.
The polysaccharide accounts for 10-20% of the total weight of the BMP protein preparation.
A medical kit for delivering a protein, comprising BMP protein incorporated into a polymeric matrix; excipients dissolved in solvents, phospholipids and polysaccharides.
The solvent is deionized water or ethanol.
A BMP protein preparation is prepared by stirring BMP protein, excipient, phospholipid and polysaccharide at 0 deg.C with a glass stirring rod.
Compared with the prior art, the invention has the following beneficial effects: the BMP protein preparation of the invention can be exposed to normal temperature air for more than 10 days in the process of orthopedic surgery treatment, ensures smooth operation, can be stored for more than 30 days at low temperature (0 ℃) and can be stored for a plurality of years at freezing condition (-80 ℃), and the activity is not changed.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.
Example 1
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol, 15 parts of lecithin and 15 parts of trehalose.
The components are stirred and mixed evenly at low temperature, the activity is lost by 8 percent within 10 days at room temperature, the activity is lost by 5 percent when the components are stored for 30 days at 0 ℃, the components are stored for 3 years at-80 ℃, and the activity is lost by 15 percent after unfreezing.
Example 2
A BMP protein preparation is composed of the following substances in parts by weight: 35 parts of BMP protein, 35 parts of gelatin, 18 parts of cephalin and 12 parts of undaria pinnatifida polysaccharide.
The components are stirred and mixed evenly at low temperature, the activity is lost by 7% within 10 days at room temperature, the activity is lost by 6% after being stored for 30 days at 0 ℃, the activity is lost by 16% after being stored for 3 years at-80 ℃.
Example 3
A BMP protein preparation is composed of the following substances in parts by weight: 25 parts of BMP protein, 42 parts of sodium starch glycolate, 18 parts of cardiolipin and 15 parts of isomaltooligosaccharide.
The components are stirred and mixed evenly at low temperature, the activity is lost by 7 percent within 10 days at room temperature, the activity is lost by 5 percent when the mixture is stored for 30 days at 0 ℃, the activity is lost by 14 percent when the mixture is stored for 3 years at-80 ℃ and unfrozen.
Example 4
A BMP protein preparation is composed of the following substances in parts by weight: 28 parts of BMP protein, 32 parts of hydroxypropyl methylcellulose, 20 parts of sphingomyelin and 20 parts of arabinogalactan.
The components are stirred and mixed evenly at low temperature, the activity is lost by 6 percent within 10 days at room temperature, the activity is lost by 6 percent when the mixture is stored for 30 days at 0 ℃, the activity is lost by 13 percent when the mixture is stored for 3 years at-80 ℃.
Example 5
A BMP protein preparation is composed of the following substances in parts by weight: 28 parts of BMP protein, 32 parts of hydroxypropyl methylcellulose, 20 parts of sphingomyelin and 20 parts of beta-1, 6-glucan.
The components are stirred and mixed evenly at low temperature, the activity is lost by 6 percent within 10 days at room temperature, the activity is lost by 6 percent when the mixture is stored for 30 days at 0 ℃, the activity is lost by 13 percent when the mixture is stored for 3 years at-80 ℃.
Example 6
A BMP protein preparation is composed of the following substances in parts by weight: 40 parts of BMP protein, 30 parts of sucrose stearate, 10 parts of phosphatidylserine and 20 parts of arabinogalactan.
The components are stirred and mixed evenly at low temperature, the activity is lost by 6 percent within 10 days at room temperature, the activity is lost by 8 percent when the components are stored for 30 days at 0 ℃, the activity is lost by 17 percent when the components are stored for 3 years at-80 ℃.
Example 7
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 20 parts of starch octenylsuccinate, 20 parts of mPEG-PLA-lysine, 10 parts of phosphatidylglycerol and 20 parts of arabinogalactan.
The components are stirred and mixed evenly at low temperature, the activity is lost by 6 percent within 10 days at room temperature, the activity is lost by 8 percent when the components are stored for 30 days at 0 ℃, the activity is lost by 17 percent when the components are stored for 3 years at-80 ℃.
Example 8
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol, 15 parts of lecithin, 15 parts of trehalose and 1 part of a beckmannia syzigachne extract.
The method for extracting the beckmannia syzigachne extract comprises the following steps: drying American Slickgrass in the sun, grinding into powder, adding 3-6 times of 70% ethanol solution, reflux extracting for 3 times, mixing filtrates, decolorizing with active carbon, and evaporating to dryness.
The components are stirred and mixed evenly at low temperature, the activity is lost by 1% within 10 days at room temperature, the activity is lost by 1% after being stored for 30 days at 0 ℃, the activity is lost by 3 years at-80 ℃, and the activity is lost by 2% after being unfrozen.
Example 9
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol, 15 parts of lecithin, 15 parts of trehalose and 1 part of a hard grass extract.
The method for extracting the beckmannia syzigachne extract comprises the following steps: sun drying radix Stephaniae Tetrandrae, grinding into powder, reflux extracting with 3-6 times of 70% ethanol solution for 3 times, mixing filtrates, decolorizing with active carbon, and evaporating to dryness.
The components are stirred and mixed evenly at low temperature, the activity is lost by 1% within 10 days at room temperature, the activity is lost by 1% after being stored for 30 days at 0 ℃, the activity is lost by 3 years at-80 ℃, and the activity is lost by 2% after being unfrozen.
Comparative example 1
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 15 parts of lecithin and 15 parts of trehalose.
The components are stirred and mixed evenly at low temperature, the activity is lost by 48 percent within 10 days at room temperature, 55 percent at 0 ℃ for 30 days, and the activity is lost by 85 percent after being stored at-80 ℃ for 3 years and unfrozen.
Example 2
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol and 15 parts of trehalose.
The components are stirred and mixed evenly at low temperature, the activity is lost by 38% within 10 days at room temperature, the activity is lost by 35% within 30 days at 0 ℃, the activity is lost by 55% after being stored for 3 years at-80 ℃.
Example 3
A BMP protein preparation is composed of the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol and 15 parts of lecithin.
The components are stirred and mixed evenly at low temperature, the activity is lost by 48 percent within 10 days at room temperature, 55 percent at 0 ℃ for 30 days, and the activity is lost by 85 percent after being stored at-80 ℃ for 3 years and unfrozen.
The above disclosure is only for the specific embodiment of the present invention, but the present invention is not limited thereto, and any variations that can be made by those skilled in the art should fall within the scope of the present invention.
Claims (2)
1. The BMP protein preparation is characterized by comprising the following substances in parts by weight: 30 parts of BMP protein, 40 parts of glycerol, 15 parts of lecithin, 15 parts of trehalose and 1 part of a beckmannia syzigachne extract;
the method for extracting the beckmannia syzigachne extract comprises the following steps: drying American Slickgrass in the sun, grinding into powder, adding 3-6 times of 70% ethanol solution, reflux extracting for 3 times, mixing filtrates, decolorizing with active carbon, and evaporating to dryness.
2. The method for preparing a BMP protein formulation of claim 1, wherein BMP protein, excipient, phospholipid, polysaccharide and the extract of beckmarka syzigachne are stirred with a glass stirring rod at 0 ℃.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610669266.4A CN106075400B (en) | 2016-08-16 | 2016-08-16 | BMP protein preparation and preparation method thereof |
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| CN201610669266.4A CN106075400B (en) | 2016-08-16 | 2016-08-16 | BMP protein preparation and preparation method thereof |
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| CN106075400B true CN106075400B (en) | 2020-09-22 |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1403153A (en) * | 2002-10-09 | 2003-03-19 | 浙江大学 | Water dispersive nano level bone morphogenetic protein injection prepn and its prepn process |
| CN101198348A (en) * | 2005-06-17 | 2008-06-11 | 伊西康公司 | Bone morphogenetic protein formulations |
| CN101745103A (en) * | 2010-01-19 | 2010-06-23 | 广东卫伦生物制药有限公司 | Normal-temperature preserved albumin preparation |
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- 2016-08-16 CN CN201610669266.4A patent/CN106075400B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1403153A (en) * | 2002-10-09 | 2003-03-19 | 浙江大学 | Water dispersive nano level bone morphogenetic protein injection prepn and its prepn process |
| CN101198348A (en) * | 2005-06-17 | 2008-06-11 | 伊西康公司 | Bone morphogenetic protein formulations |
| CN101745103A (en) * | 2010-01-19 | 2010-06-23 | 广东卫伦生物制药有限公司 | Normal-temperature preserved albumin preparation |
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| CN106075400A (en) | 2016-11-09 |
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