CN108785661A - A kind of injection and preparation method thereof for the treatment of degenerative osteoarthropathy without side-effects - Google Patents
A kind of injection and preparation method thereof for the treatment of degenerative osteoarthropathy without side-effects Download PDFInfo
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- 238000002347 injection Methods 0.000 title claims abstract description 48
- 239000007924 injection Substances 0.000 title claims abstract description 48
- 230000000694 effects Effects 0.000 title claims abstract description 32
- 206010057178 Osteoarthropathies Diseases 0.000 title claims abstract description 27
- 230000003412 degenerative effect Effects 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 54
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 42
- 229920001661 Chitosan Polymers 0.000 claims abstract description 39
- 229920002125 Sokalan® Polymers 0.000 claims abstract description 38
- 239000004584 polyacrylic acid Substances 0.000 claims abstract description 38
- 229920000728 polyester Polymers 0.000 claims abstract description 33
- 229920000642 polymer Polymers 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 33
- 239000002904 solvent Substances 0.000 claims abstract description 26
- 210000000130 stem cell Anatomy 0.000 claims abstract description 26
- 239000000725 suspension Substances 0.000 claims abstract description 25
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000004472 Lysine Substances 0.000 claims abstract description 23
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims abstract description 23
- 229910052791 calcium Inorganic materials 0.000 claims abstract description 23
- 239000011575 calcium Substances 0.000 claims abstract description 23
- 239000000470 constituent Substances 0.000 claims abstract description 22
- 102000008186 Collagen Human genes 0.000 claims abstract description 20
- 108010035532 Collagen Proteins 0.000 claims abstract description 20
- 229920001436 collagen Polymers 0.000 claims abstract description 20
- 239000003208 petroleum Substances 0.000 claims abstract description 20
- 102000008100 Human Serum Albumin Human genes 0.000 claims abstract description 18
- 108091006905 Human Serum Albumin Proteins 0.000 claims abstract description 18
- 239000008367 deionised water Substances 0.000 claims abstract description 17
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 17
- 229920002401 polyacrylamide Polymers 0.000 claims abstract description 17
- 239000003094 microcapsule Substances 0.000 claims abstract description 16
- SPTHWAJJMLCAQF-UHFFFAOYSA-N 1,2-di(propan-2-yl)benzene;hydrogen peroxide Chemical compound OO.CC(C)C1=CC=CC=C1C(C)C SPTHWAJJMLCAQF-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000004475 Arginine Substances 0.000 claims abstract description 15
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000005406 washing Methods 0.000 claims abstract description 13
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 229940090044 injection Drugs 0.000 claims description 43
- 239000007864 aqueous solution Substances 0.000 claims description 24
- 238000005119 centrifugation Methods 0.000 claims description 24
- 239000006228 supernatant Substances 0.000 claims description 19
- 238000003756 stirring Methods 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 229920001218 Pullulan Polymers 0.000 claims description 12
- 239000004373 Pullulan Substances 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 235000019423 pullulan Nutrition 0.000 claims description 12
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 10
- RWGFKTVRMDUZSP-UHFFFAOYSA-N cumene Chemical compound CC(C)C1=CC=CC=C1 RWGFKTVRMDUZSP-UHFFFAOYSA-N 0.000 claims description 8
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 6
- 239000002202 Polyethylene glycol Substances 0.000 claims description 6
- 229920000954 Polyglycolide Polymers 0.000 claims description 6
- WNLRTRBMVRJNCN-UHFFFAOYSA-L adipate(2-) Chemical compound [O-]C(=O)CCCCC([O-])=O WNLRTRBMVRJNCN-UHFFFAOYSA-L 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000004945 emulsification Methods 0.000 claims description 6
- 235000019441 ethanol Nutrition 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 229920001223 polyethylene glycol Polymers 0.000 claims description 6
- 239000004633 polyglycolic acid Substances 0.000 claims description 6
- 239000013049 sediment Substances 0.000 claims description 6
- 238000013517 stratification Methods 0.000 claims description 6
- 102000004452 Arginase Human genes 0.000 claims description 4
- 108700024123 Arginases Proteins 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- 239000003792 electrolyte Substances 0.000 claims description 4
- 229940093181 glucose injection Drugs 0.000 claims description 4
- 150000004676 glycans Chemical class 0.000 claims 2
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims 1
- 238000002242 deionisation method Methods 0.000 claims 1
- 239000003292 glue Substances 0.000 claims 1
- 230000001225 therapeutic effect Effects 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 230000004069 differentiation Effects 0.000 abstract description 3
- 230000001900 immune effect Effects 0.000 abstract description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 230000002062 proliferating effect Effects 0.000 abstract description 3
- 230000005847 immunogenicity Effects 0.000 abstract description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 8
- OKIRBHVFJGXOIS-UHFFFAOYSA-N 1,2-di(propan-2-yl)benzene Chemical compound CC(C)C1=CC=CC=C1C(C)C OKIRBHVFJGXOIS-UHFFFAOYSA-N 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 239000002775 capsule Substances 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 239000002253 acid Substances 0.000 description 4
- 229910021529 ammonia Inorganic materials 0.000 description 4
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 3
- 208000032170 Congenital Abnormalities Diseases 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 201000008482 osteoarthritis Diseases 0.000 description 2
- ODLMAHJVESYWTB-UHFFFAOYSA-N propylbenzene Chemical compound CCCC1=CC=CC=C1 ODLMAHJVESYWTB-UHFFFAOYSA-N 0.000 description 2
- 208000006820 Arthralgia Diseases 0.000 description 1
- 206010023232 Joint swelling Diseases 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 150000001408 amides Chemical group 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007698 birth defect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000004394 hip joint Anatomy 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 210000001503 joint Anatomy 0.000 description 1
- 210000000629 knee joint Anatomy 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- -1 polypropylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000005065 subchondral bone plate Anatomy 0.000 description 1
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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
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
- A61K31/722—Chitin, chitosan
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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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
- A61K38/38—Albumins
- A61K38/385—Serum albumin
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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
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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Abstract
The invention discloses a kind of injections for the treatment of degenerative osteoarthropathy without side-effects, including following component：Person joint's liquid mescenchymal stem cell 22-28%, human serum albumin 6.8-7.6%, calcium constituent 5.25-6.45%, polyacrylic acid 3.6-4.3%, di-isopropylbenzene hydroperoxide 2.8-4.5%, polyester polymers 2.1-2.9%, polyacrylamide 1.8-2.2%, chitosan 2.33-2.87%, petroleum ether 1.24-2.24%, arginine 1.5-2.2%, lysine 1.1-1.7%, collagen 0.8-1.3%, solvent 4.8-6.3%, deionized water are surplus；The invention also discloses a kind of preparation methods of the injection for the treatment of degenerative osteoarthropathy without side-effects, include the following steps：Suspension is prepared, polyacrylic acid crosslinked object is prepared, prepares chitosan microcapsules, washing, centrifugal filtration and obtained injection；The formula of the present invention is more reasonable, a large amount of person joint's liquid mescenchymal stem cell is being prepared in the present invention, it is prepared into injection, it is big with differentiation potential, proliferative capacity is strong, the low advantage of immunogenicity, can be good at the generation for inhibiting immunological rejection, therapeutic effect is good, without side-effects.
Description
Technical field
The present invention relates to field of biomedicine technology, specially a kind of injection for the treatment of degenerative osteoarthropathy without side-effects
Agent and preparation method thereof.
Background technology
Degenerative osteoarthropathy is also known as osteoarthritis, degenerative arthritis, senescent arthritis, hypertrophiarthritis, it
It is a kind of retrogression pathological changes, is due to increasing the factors such as age, obesity, strain, wound, joint birth defect, joint deformity
Caused articular cartilage degeneration damage, joint margins and subchondral bone reactive hyperplasia.This disease is more common in mid-aged population, good to send out
In the more joint of weight-bearing joints and activity (e.g., cervical vertebra, lumbar vertebrae, knee joint, hip joint etc.).It excessively bears a heavy burden or uses these
Joint, the generation that can promote degeneration to change.Clinical manifestation is the arthralgia, tenderness, stiff, swollen joint slowly developed
Swollen, limitation of activity and joint deformity etc., degenerative osteoarthropathy have seriously affected the physical and mental health of the middle-aged and the old, but traditional
Treatment degenerative osteoarthropathy injection generally existing therapeutic effect it is poor, user's rejection is stronger, and side effect is big
Defect；For this purpose, a kind of injection for the treatment of degenerative osteoarthropathy without side-effects of proposition and preparation method thereof is necessary.
Invention content
The purpose of the present invention is to provide a kind of injection for the treatment of degenerative osteoarthropathy without side-effects and its preparation sides
Method, to solve the problems, such as to propose in background technology.
To achieve the above object, the present invention provides the following technical solutions：A kind for the treatment of degenerative osteoarthropathy without side-effects
Injection, including following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 22-28%, human serum albumin
6.8-7.6%, calcium constituent 5.25-6.45%, polyacrylic acid 3.6-4.3%, di-isopropylbenzene hydroperoxide 2.8-4.5%, polyester
Polymer 2.1-2.9%, polyacrylamide 1.8-2.2%, chitosan 2.33-2.87%, petroleum ether 1.24-2.24%, smart ammonia
Sour 1.5-2.2%, lysine 1.1-1.7%, collagen 0.8-1.3%, solvent 4.8-6.3%, deionized water are surplus.
Preferably, including following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 22%, the white egg of people's blood
White 6.8%, calcium constituent 5.25%, polyacrylic acid 3.6%, di-isopropylbenzene hydroperoxide 2.8%, polyester polymers 2.1%, poly- third
Acrylamide 1.8%, chitosan 2.33%, petroleum ether 1.24%, arginine 1.5%, lysine 1.1%, collagen 0.8%,
Solvent 4.8%, deionized water are surplus.
Preferably, including following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 24%, the white egg of people's blood
White 7.2%, calcium constituent 5.69%, polyacrylic acid 3.8%, di-isopropylbenzene hydroperoxide 3%, polyester polymers 2.4%, polypropylene
It is amide 1.9%, chitosan 2.55%, petroleum ether 1.62%, arginine 1.8%, lysine 1.4%, collagen 0.9%, molten
Matchmaker 5.2%, deionized water are surplus.
Preferably, including following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 26%, the white egg of people's blood
White 7.4%, calcium constituent 6.12%, polyacrylic acid 4.1%, di-isopropylbenzene hydroperoxide 4.3%, polyester polymers 2.65%, poly-
Acrylamide 2.1%, chitosan 2.74%, petroleum ether 1.89%, arginase 12 .1%, lysine 1.6%, collagen
1.1%, solvent 5.8%, deionized water are surplus.
Preferably, including following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 28%, the white egg of people's blood
White 7.6%, calcium constituent 6.45%, polyacrylic acid 4.3%, di-isopropylbenzene hydroperoxide 4.5%, polyester polymers 2.9%, poly- third
Acrylamide 2.2%, chitosan 2.87%, petroleum ether 2.24%, arginase 12 .2%, lysine 1.7%, collagen 1.3%,
Solvent 6.3%, deionized water are surplus.
Preferably, the polyester polymers are made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and three
Ratio be 1:1:2；The solvent is any one of Multiple electrolytes injection or glucose injection.
A kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, is made
Polyester polymers aqueous solution；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 6.5-8.5g/mL；By poly- third
Olefin(e) acid is added in ethyl alcohol and forms oil phase, and pullulan aqueous solution is added in oil phase, then peroxidating is added in clipped machine emulsification
Hydrogen diisopropylbenzene (DIPB), is stirred at room temperature, and crosslinks reaction 70-130 minutes, then stratification is dried to get polyacrylic acid
Cross-linking agent；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 65-75 C water baths to heat and stir 20-
30 minutes, chitosan is then added, it is that 2000-3000 turns/min to improve rotating speed, obtains mixture, oil is added into mixture
Ether, continues to stir, be then filtered by filter, and the chitosan microcapsules that grain size is 15-30 μm are obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 2-5 times using PBS liquid, then uses horizontal centrifugal
Machine centrifuges 30-40 minutes under conditions of 21-25 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object is sequentially added into the suspension after washing and chitosan is micro-
Then solvent is added in capsule, stirred evenly under conditions of 25-45 degrees Celsius, then passes through centrifugation apparatus and centrifuges 10-20 minutes,
And supernatant liquid is detached, obtain mixed liquor A；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into
In high-speed mixing equipment, and deionized water is added, control rotating speed is that 1200-1600 turns/min is mixed 15-30 minutes, stirring
Shi Wendu is 35-45 degrees Celsius, is then put into centrifugation apparatus centrifugal filtration 5-10 minutes, removes supernatant and waits for its nature
It is cooling, obtain mixed liquid B；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, centrifugation is abandoned
Clear liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
Compared with prior art, the beneficial effects of the invention are as follows：The formula of the present invention is more reasonable, prepared by the present invention
A large amount of person joint's liquid mescenchymal stem cell is obtained, injection is prepared into, passes through addition human serum albumin, calcium in injection
Element required for a variety of human synovials of element, also cooperation addition chitosan microcapsules, arginine and lysine so that joint mesenchyma
Stem cell obtains preferable active protection, provides cell required energy, big with differentiation potential, proliferative capacity is strong, exempts from
The low advantage of epidemic focus can be good at the generation for inhibiting immunological rejection, and therapeutic effect is good, without side-effects；And this hair
Bright preparation method convenient material drawing, equipment requirement is low, has good promotion effect.
Specific implementation mode
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Embodiment one：
A kind of injection for the treatment of degenerative osteoarthropathy without side-effects, including following component (by mass percentage)：
Person joint's liquid mescenchymal stem cell 22%, human serum albumin 6.8%, calcium constituent 5.25%, polyacrylic acid 3.6%, hydrogen peroxide
Diisopropylbenzene (DIPB) 2.8%, polyester polymers 2.1%, polyacrylamide 1.8%, chitosan 2.33%, petroleum ether 1.24%, smart ammonia
Acid 1.5%, lysine 1.1%, collagen 0.8%, solvent 4.8%, deionized water are surplus.
Further, polyester polymers are made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and three
Ratio is 1:1:2；Solvent is Multiple electrolytes injection.
A kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, is made
Polyester polymers aqueous solution；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 6.5g/mL；By polyacrylic acid
It is added in ethyl alcohol and forms oil phase, pullulan aqueous solution is added in oil phase, then hydrogen peroxide two is added in clipped machine emulsification
Isopropylbenzene is stirred at room temperature, and crosslinks reaction 70 minutes, then stratification is dried to get polyacrylic acid crosslinked object；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 65 C water baths to heat and stir 20 minutes,
Then chitosan is added, raising rotating speed is 2000 turns/min, obtains mixture, petroleum ether is added into mixture, continue to stir,
Then it is filtered by filter, the chitosan microcapsules that grain size is 15 μm is obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 2 times using PBS liquid, then uses horizontal centrifuge
It is centrifuged 30 minutes under conditions of 21 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object is sequentially added into the suspension after washing and chitosan is micro-
Then solvent is added in capsule, stirred evenly under conditions of 25 degrees Celsius, then passes through centrifugation apparatus and centrifuges 10 minutes, and detaches
Supernatant liquid obtains mixed liquor A；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into
In high-speed mixing equipment, and deionized water is added, control rotating speed is that 1200 turns/min is mixed 15 minutes, and temperature is when stirring
35 degrees Celsius, centrifugal filtration 5 minutes in centrifugation apparatus are then put into, remove supernatant and wait for its natural cooling, are mixed
Liquid B；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, centrifugation is abandoned
Clear liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
Embodiment two：
A kind of injection for the treatment of degenerative osteoarthropathy without side-effects, including following component (by mass percentage)：
Person joint's liquid mescenchymal stem cell 24%, human serum albumin 7.2%, calcium constituent 5.69%, polyacrylic acid 3.8%, hydrogen peroxide
Diisopropylbenzene (DIPB) 3%, polyester polymers 2.4%, polyacrylamide 1.9%, chitosan 2.55%, petroleum ether 1.62%, arginine
1.8%, lysine 1.4%, collagen 0.9%, solvent 5.2%, deionized water are surplus.
Further, polyester polymers are made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and three
Ratio is 1:1:2；Solvent is glucose injection.
A kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, is made
Polyester polymers aqueous solution；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 7.5g/mL；By polyacrylic acid
It is added in ethyl alcohol and forms oil phase, pullulan aqueous solution is added in oil phase, then hydrogen peroxide two is added in clipped machine emulsification
Isopropylbenzene is stirred at room temperature, and crosslinks reaction 90 minutes, then stratification is dried to get polyacrylic acid crosslinked object；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 68 C water baths to heat and stir 24 minutes,
Then chitosan is added, raising rotating speed is 2300 turns/min, obtains mixture, petroleum ether is added into mixture, continue to stir,
Then it is filtered by filter, the chitosan microcapsules that grain size is 23 μm is obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 3 times using PBS liquid, then uses horizontal centrifuge
It is centrifuged 34 minutes under conditions of 22 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object is sequentially added into the suspension after washing and chitosan is micro-
Then solvent is added in capsule, stirred evenly under conditions of 35 degrees Celsius, then passes through centrifugation apparatus and centrifuges 12 minutes, and detaches
Supernatant liquid obtains mixed liquor A；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into
In high-speed mixing equipment, and deionized water is added, control rotating speed is that 1300 turns/min is mixed 20 minutes, and temperature is when stirring
37 degrees Celsius, centrifugal filtration 7 minutes in centrifugation apparatus are then put into, remove supernatant and wait for its natural cooling, are mixed
Liquid B；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, centrifugation is abandoned
Clear liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
Embodiment three：
A kind of injection for the treatment of degenerative osteoarthropathy without side-effects, including following component (by mass percentage)：
Person joint's liquid mescenchymal stem cell 26%, human serum albumin 7.4%, calcium constituent 6.12%, polyacrylic acid 4.1%, hydrogen peroxide
Diisopropylbenzene (DIPB) 4.3%, polyester polymers 2.65%, polyacrylamide 2.1%, chitosan 2.74%, petroleum ether 1.89%, smart ammonia
Acid 2.1%, lysine 1.6%, collagen 1.1%, solvent 5.8%, deionized water are surplus.
Further, polyester polymers are made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and three
Ratio is 1:1:2；Solvent is Multiple electrolytes injection.
A kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, is made
Polyester polymers aqueous solution；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 8g/mL；By polyacrylic acid plus
Enter and form oil phase in ethyl alcohol, pullulan aqueous solution is added in oil phase, then it is different that hydrogen peroxide two is added in clipped machine emulsification
Propyl benzene is stirred at room temperature, and crosslinks reaction 110 minutes, then stratification is dried to get polyacrylic acid crosslinked object；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 72 C water baths to heat and stir 27 minutes,
Then chitosan is added, raising rotating speed is 2700 turns/min, obtains mixture, petroleum ether is added into mixture, continue to stir,
Then it is filtered by filter, the chitosan microcapsules that grain size is 25 μm is obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 4 times using PBS liquid, then uses horizontal centrifuge
It is centrifuged 37 minutes under conditions of 24 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object is sequentially added into the suspension after washing and chitosan is micro-
Then solvent is added in capsule, stirred evenly under conditions of 40 degrees Celsius, then passes through centrifugation apparatus and centrifuges 17 minutes, and detaches
Supernatant liquid obtains mixed liquor A；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into
In high-speed mixing equipment, and deionized water is added, control rotating speed is that 1500 turns/min is mixed 25 minutes, and temperature is when stirring
42 degrees Celsius, centrifugal filtration 8 minutes in centrifugation apparatus are then put into, remove supernatant and wait for its natural cooling, are mixed
Liquid B；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, centrifugation is abandoned
Clear liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
Example IV：
A kind of injection for the treatment of degenerative osteoarthropathy without side-effects, including following component (by mass percentage)：
Person joint's liquid mescenchymal stem cell 28%, human serum albumin 7.6%, calcium constituent 6.45%, polyacrylic acid 4.3%, hydrogen peroxide
Diisopropylbenzene (DIPB) 4.5%, polyester polymers 2.9%, polyacrylamide 2.2%, chitosan 2.87%, petroleum ether 2.24%, smart ammonia
Acid 2.2%, lysine 1.7%, collagen 1.3%, solvent 6.3%, deionized water are surplus.
Further, polyester polymers are made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and three
Ratio is 1:1:2；Solvent is glucose injection.
A kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, is made
Polyester polymers aqueous solution；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 8.5g/mL；By polyacrylic acid
It is added in ethyl alcohol and forms oil phase, pullulan aqueous solution is added in oil phase, then hydrogen peroxide two is added in clipped machine emulsification
Isopropylbenzene is stirred at room temperature, and crosslinks reaction 130 minutes, then stratification is dried to get polyacrylic acid crosslinked object；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 75 C water baths to heat and stir 30 minutes,
Then chitosan is added, raising rotating speed is 3000 turns/min, obtains mixture, petroleum ether is added into mixture, continue to stir,
Then it is filtered by filter, the chitosan microcapsules that grain size is 30 μm is obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 5 times using PBS liquid, then uses horizontal centrifuge
It is centrifuged 40 minutes under conditions of 25 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object is sequentially added into the suspension after washing and chitosan is micro-
Then solvent is added in capsule, stirred evenly under conditions of 45 degrees Celsius, then passes through centrifugation apparatus and centrifuges 20 minutes, and detaches
Supernatant liquid obtains mixed liquor A；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into
In high-speed mixing equipment, and deionized water is added, control rotating speed is that 1600 turns/min is mixed 30 minutes, and temperature is when stirring
45 degrees Celsius, centrifugal filtration 10 minutes in centrifugation apparatus are then put into, remove supernatant and wait for its natural cooling, are mixed
Liquid B；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, centrifugation is abandoned
Clear liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
Injection can be made by above four groups of embodiments, and the formula of the present invention is more reasonable, the present invention exists
A large amount of person joint's liquid mescenchymal stem cell is prepared, is prepared into injection, passes through the addition white egg of people's blood in injection
In vain, element required for a variety of human synovials of calcium constituent, also cooperation addition chitosan microcapsules, arginine and lysine so that joint
Mescenchymal stem cell obtains preferable active protection, provides cell required energy, proliferative capacity big with differentiation potential
By force, the low advantage of immunogenicity can be good at the generation for inhibiting immunological rejection, and therapeutic effect is good, without side-effects；And
The preparation method convenient material drawing of the present invention, equipment requirement is low, has good promotion effect.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Claims (7)
1. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects, which is characterized in that (press quality hundred including following component
Divide than meter)：Person joint's liquid mescenchymal stem cell 22-28%, human serum albumin 6.8-7.6%, calcium constituent 5.25-6.45%, gather
Acrylic acid 3.6-4.3%, di-isopropylbenzene hydroperoxide 2.8-4.5%, polyester polymers 2.1-2.9%, polyacrylamide 1.8-
2.2%, chitosan 2.33-2.87%, petroleum ether 1.24-2.24%, arginine 1.5-2.2%, lysine 1.1-1.7%, glue
Former albumen 0.8-1.3%, solvent 4.8-6.3%, deionized water are surplus.
2. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects according to claim 1, which is characterized in that packet
Include following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 22%, human serum albumin 6.8%, calcium constituent
5.25%, polyacrylic acid 3.6%, di-isopropylbenzene hydroperoxide 2.8%, polyester polymers 2.1%, polyacrylamide 1.8%, shell
Glycan 2.33%, petroleum ether 1.24%, arginine 1.5%, lysine 1.1%, collagen 0.8%, solvent 4.8%, go from
Sub- water is surplus.
3. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects according to claim 1, which is characterized in that packet
Include following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 24%, human serum albumin 7.2%, calcium constituent
5.69%, polyacrylic acid 3.8%, di-isopropylbenzene hydroperoxide 3%, polyester polymers 2.4%, polyacrylamide 1.9%, shell are poly-
Sugar 2.55%, petroleum ether 1.62%, arginine 1.8%, lysine 1.4%, collagen 0.9%, solvent 5.2%, deionization
Water is surplus.
4. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects according to claim 1, which is characterized in that packet
Include following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 26%, human serum albumin 7.4%, calcium constituent
6.12%, polyacrylic acid 4.1%, di-isopropylbenzene hydroperoxide 4.3%, polyester polymers 2.65%, polyacrylamide 2.1%,
Chitosan 2.74%, arginase 12 .1%, lysine 1.6%, collagen 1.1%, solvent 5.8%, is gone petroleum ether 1.89%
Ionized water is surplus.
5. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects according to claim 1, which is characterized in that packet
Include following component (by mass percentage)：Person joint's liquid mescenchymal stem cell 28%, human serum albumin 7.6%, calcium constituent
6.45%, polyacrylic acid 4.3%, di-isopropylbenzene hydroperoxide 4.5%, polyester polymers 2.9%, polyacrylamide 2.2%, shell
Glycan 2.87%, petroleum ether 2.24%, arginase 12 .2%, lysine 1.7%, collagen 1.3%, solvent 6.3%, go from
Sub- water is surplus.
6. a kind of injection for the treatment of degenerative osteoarthropathy without side-effects according to claim 1, it is characterised in that：Institute
It states polyester polymers to be made of polyglycolic acid, pla-pcl and polyethylene glycol adipate, and the ratio of three is 1:1:2；Institute
State any one that solvent is Multiple electrolytes injection or glucose injection.
7. a kind of preparation method of the injection for the treatment of degenerative osteoarthropathy without side-effects as described in claim 1-6,
It is characterized in that, includes the following steps：
Step 1：Prepare suspension：Polyester polymers are dissolved in the water of 1.5 parts by weight, and be mixed evenly, polyester is made
Aqueous solutions of polymers；Mescenchymal stem cell is suspended in aqueous solution, suspension is made；
Step 2：Prepare polyacrylic acid crosslinked object：Configuration concentration is the pullulan aqueous solution of 6.5-8.5g/mL；By polyacrylic acid
It is added in ethyl alcohol and forms oil phase, pullulan aqueous solution is added in oil phase, then hydrogen peroxide two is added in clipped machine emulsification
Isopropylbenzene is stirred at room temperature, and crosslinks reaction 70-130 minutes, then stratification is dried to get polyacrylic acid crosslinked
Object；
Step 3：Prepare chitosan microcapsules：It takes polyacrylamide to be placed in 65-75 C water baths and heats and stir 20-30 points
Then chitosan is added in clock, it is that 2000-3000 turns/min to improve rotating speed, obtains mixture, petroleum ether is added into mixture,
Continue to stir, be then filtered by filter, the chitosan microcapsules that grain size is 15-30 μm are obtained after natural drying；
Step 4：Washing：It takes the suspension obtained in step 1 to be washed 2-5 times using PBS liquid, is then existed using horizontal centrifuge
It is centrifuged 30-40 minutes under conditions of 21-25 degrees Celsius；
Step 5：Prepare mixed liquor A：Polyacrylic acid crosslinked object and chitosan microcapsules are sequentially added into the suspension after washing,
Then solvent is added, is stirred evenly under conditions of 25-45 degrees Celsius, then passes through centrifugation apparatus and centrifuges 10-20 minutes, and point
From supernatant liquid, mixed liquor A is obtained；
Step 6：Prepare mixed liquid B：Human serum albumin, calcium constituent, arginine, lysine and collagen is taken to be put into high speed
In mixing plant, and deionized water is added, control rotating speed be 1200-1600 turn/min is mixed 15-30 minutes, temperature when stirring
Degree is 35-45 degrees Celsius, is then put into centrifugation apparatus centrifugal filtration 5-10 minutes, removes supernatant and waits for that it is naturally cold
But, mixed liquid B is obtained；
Step 7：Injection is made：Mixed liquid B and mixed liquor A are merged and stirred evenly under normal temperature condition, supernatant is abandoned in centrifugation
Liquid, sediment are resuspended with DMEM/F12 culture mediums, and supernatant is abandoned in centrifugation again, is finally frozen spare, you can obtain injection.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110237095A (en) * | 2019-05-28 | 2019-09-17 | 武汉汉密顿生物科技股份有限公司 | For treating stem cell injection liquid and its preparation and the application of cartilage defect of osteoarthritis |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160058867A1 (en) * | 2013-04-09 | 2016-03-03 | The Board Of Trustees Of The Leland Stanford Junior University | Crosslinked chitosan-lactide hydrogels |
| WO2017212057A1 (en) * | 2016-06-10 | 2017-12-14 | Regulaxis | Peptides for the treatment of osteoarthritis |
| CN107898810A (en) * | 2017-12-20 | 2018-04-13 | 北京臻惠康生物科技有限公司 | A kind of mescenchymal stem cell repairs parenteral solution and preparation method thereof |
-
2018
- 2018-07-20 CN CN201810803938.5A patent/CN108785661A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160058867A1 (en) * | 2013-04-09 | 2016-03-03 | The Board Of Trustees Of The Leland Stanford Junior University | Crosslinked chitosan-lactide hydrogels |
| WO2017212057A1 (en) * | 2016-06-10 | 2017-12-14 | Regulaxis | Peptides for the treatment of osteoarthritis |
| CN107898810A (en) * | 2017-12-20 | 2018-04-13 | 北京臻惠康生物科技有限公司 | A kind of mescenchymal stem cell repairs parenteral solution and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110237095A (en) * | 2019-05-28 | 2019-09-17 | 武汉汉密顿生物科技股份有限公司 | For treating stem cell injection liquid and its preparation and the application of cartilage defect of osteoarthritis |
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