CN103191153B - Muscular amino acid and nucleoside extract and pharmaceutical composition thereof - Google Patents
Muscular amino acid and nucleoside extract and pharmaceutical composition thereof Download PDFInfo
- Publication number
- CN103191153B CN103191153B CN201310153538.1A CN201310153538A CN103191153B CN 103191153 B CN103191153 B CN 103191153B CN 201310153538 A CN201310153538 A CN 201310153538A CN 103191153 B CN103191153 B CN 103191153B
- Authority
- CN
- China
- Prior art keywords
- homogenate
- sarcosine peptide
- peptide aglycone
- hours
- ultrasonic field
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000284 extract Substances 0.000 title claims abstract description 31
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 20
- 150000001413 amino acids Chemical class 0.000 title abstract description 14
- 239000002777 nucleoside Substances 0.000 title abstract description 13
- 230000003387 muscular Effects 0.000 title abstract description 12
- RYSMHWILUNYBFW-GRIPGOBMSA-N 3'-amino-3'-deoxy-N(6),N(6)-dimethyladenosine Chemical compound C1=NC=2C(N(C)C)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](N)[C@H]1O RYSMHWILUNYBFW-GRIPGOBMSA-N 0.000 title abstract 10
- 239000000243 solution Substances 0.000 claims abstract description 45
- 238000002347 injection Methods 0.000 claims abstract description 43
- 239000007924 injection Substances 0.000 claims abstract description 43
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 claims abstract description 40
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000000203 mixture Substances 0.000 claims abstract description 23
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 22
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 claims abstract description 20
- PJUIMOJAAPLTRJ-UHFFFAOYSA-N monothioglycerol Chemical compound OCC(O)CS PJUIMOJAAPLTRJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229940035024 thioglycerol Drugs 0.000 claims abstract description 18
- 241000283973 Oryctolagus cuniculus Species 0.000 claims abstract description 14
- 210000003205 muscle Anatomy 0.000 claims abstract description 14
- ODLHGICHYURWBS-LKONHMLTSA-N trappsol cyclo Chemical compound CC(O)COC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)COCC(O)C)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1COCC(C)O ODLHGICHYURWBS-LKONHMLTSA-N 0.000 claims abstract description 14
- 210000004165 myocardium Anatomy 0.000 claims abstract description 11
- 239000001103 potassium chloride Substances 0.000 claims abstract description 11
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 11
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 11
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims abstract description 9
- 210000002808 connective tissue Anatomy 0.000 claims abstract description 9
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 claims abstract description 9
- 239000012528 membrane Substances 0.000 claims abstract description 9
- 239000000600 sorbitol Substances 0.000 claims abstract description 9
- 229940119744 dextran 40 Drugs 0.000 claims abstract description 7
- 108010077895 Sarcosine Proteins 0.000 claims description 108
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 claims description 101
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 claims description 101
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 claims description 101
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 48
- 238000003756 stirring Methods 0.000 claims description 46
- 238000002360 preparation method Methods 0.000 claims description 22
- 239000003814 drug Substances 0.000 claims description 18
- 210000001519 tissue Anatomy 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 17
- 238000002156 mixing Methods 0.000 claims description 17
- 239000008215 water for injection Substances 0.000 claims description 17
- 238000005374 membrane filtration Methods 0.000 claims description 16
- 229920001983 poloxamer Polymers 0.000 claims description 16
- 239000000706 filtrate Substances 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 10
- 238000007710 freezing Methods 0.000 claims description 8
- 230000008014 freezing Effects 0.000 claims description 8
- 238000005360 mashing Methods 0.000 claims description 8
- 238000010792 warming Methods 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 238000003556 assay Methods 0.000 claims description 4
- 238000004821 distillation Methods 0.000 claims description 4
- 230000009191 jumping Effects 0.000 claims description 4
- 238000004806 packaging method and process Methods 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 claims description 3
- 229920002307 Dextran Polymers 0.000 claims description 2
- 229920001184 polypeptide Polymers 0.000 abstract description 18
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 8
- 238000001914 filtration Methods 0.000 abstract description 4
- 150000003833 nucleoside derivatives Chemical class 0.000 abstract description 3
- 239000002773 nucleotide Substances 0.000 abstract description 3
- 125000003729 nucleotide group Chemical group 0.000 abstract description 3
- CTKXFMQHOOWWEB-UHFFFAOYSA-N Ethylene oxide/propylene oxide copolymer Chemical compound CCCOC(C)COCCO CTKXFMQHOOWWEB-UHFFFAOYSA-N 0.000 abstract 2
- 239000008176 lyophilized powder Substances 0.000 abstract 2
- 229920001993 poloxamer 188 Polymers 0.000 abstract 2
- 229940044519 poloxamer 188 Drugs 0.000 abstract 2
- 238000013019 agitation Methods 0.000 abstract 1
- 239000002245 particle Substances 0.000 abstract 1
- 238000009210 therapy by ultrasound Methods 0.000 abstract 1
- 238000000034 method Methods 0.000 description 19
- 238000012360 testing method Methods 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 7
- 229930182470 glycoside Natural products 0.000 description 7
- 150000002338 glycosides Chemical class 0.000 description 7
- 230000003301 hydrolyzing effect Effects 0.000 description 7
- 238000000605 extraction Methods 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 5
- 235000013372 meat Nutrition 0.000 description 5
- 239000013618 particulate matter Substances 0.000 description 5
- 238000011835 investigation Methods 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 206010002027 Amyotrophy Diseases 0.000 description 2
- 206010008190 Cerebrovascular accident Diseases 0.000 description 2
- 208000006011 Stroke Diseases 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 206010003549 asthenia Diseases 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 208000026106 cerebrovascular disease Diseases 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011981 development test Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 210000005003 heart tissue Anatomy 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 1
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 102000002723 Atrial Natriuretic Factor Human genes 0.000 description 1
- 101800001288 Atrial natriuretic factor Proteins 0.000 description 1
- 101800001890 Atrial natriuretic peptide Proteins 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 102000004414 Calcitonin Gene-Related Peptide Human genes 0.000 description 1
- 108090000932 Calcitonin Gene-Related Peptide Proteins 0.000 description 1
- 206010007559 Cardiac failure congestive Diseases 0.000 description 1
- 206010065559 Cerebral arteriosclerosis Diseases 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 206010019468 Hemiplegia Diseases 0.000 description 1
- 208000032456 Hemorrhagic Shock Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 208000007443 Neurasthenia Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 206010040070 Septic Shock Diseases 0.000 description 1
- 206010049771 Shock haemorrhagic Diseases 0.000 description 1
- 206010044248 Toxic shock syndrome Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- NSQLIUXCMFBZME-MPVJKSABSA-N carperitide Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CSSC[C@@H](C(=O)N1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)=O)[C@@H](C)CC)C1=CC=CC=C1 NSQLIUXCMFBZME-MPVJKSABSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000007731 hot pressing Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 201000005851 intracranial arteriosclerosis Diseases 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to a muscular amino acid and nucleoside extract containing polypeptide, amino acid, nucleoside, nucleotide and other medicinal components. The extract is prepared by crushing cardiac muscles and muscles of a rabbit of which the connective tissue is removed to be mixed with water, homogenizing, freeing and performing ultrasonic treatment; adding potassium chloride into the homogenate, and performing ultrasonic agitation; and centrifuging to obtain supernate, filtering with a filter membrane, and performing ultrafiltration. The invention also provides a muscular amino acid and nucleoside medicinal composition injection and lyophilized powder, wherein the muscular amino acid and nucleoside medicinal composition consists of the following components: 1000ml of muscular amino acid and nucleoside solution, 1.5g of sorbitol, 0.2g of thioglycerol, and 1.0g of poloxamer 188; and the concentration of the polypeptide in the muscular amino acid and nucleoside solution is between 1.58 and 1.92mg/ml, and the concentration of hypoxanthine is between 0.22 and 0.28mg/ml. The muscular amino acid and nucleoside pharmaceutical composition lyophilized powder is prepared by lyophilizing 1,000ml of muscular amino acid and nucleoside solution, 40g of dextran 40, 0.2g of thioglycerol, 1.0g of poloxamer 188, and 0.5g of hydroxypropyl betadex. The muscular amino acid and nucleoside extract and the pharmaceutical composition thereof have a good effect of improving the stability of muscular amino acid and nucleoside and maintaining low insoluble particles.
Description
Technical field
The invention belongs to medical technical field, relate to sarcosine peptide aglycone extract and pharmaceutical composition thereof.
Background technology
Sarcosine peptide aglycone be extracted by healthy rabbits muscle and cardiac muscle contain nucleotide (nucleoside) classes such as atrial natriuretic peptide, calcitonin-gene-related peptide and adenosine triphosphate (adenosine), the medicine of 20 several amino acids and metabolic intermediate isoreactivity material thereof.There is the effect of the organism metabolism of promotion.For amyotrophy, nervous edema, cerebrovascular accident paralysis, nerve asthenia syndrome etc.Ischemic cardiac, cerebrovascular, diabetes, microvascular disease.Congestive heart failure.Various types of hypertension and complication thereof.The hemiplegia that cerebrovascular accident causes, cerebral arteriosclerosis, amyotrophy, neurasthenia syndrome.Control angina pectoris, the match of cardiac muscle stalk.Toxic and hemorrhagic shock.
Prior art generally adopts multigelation to increase temperature for the extraction of sarcosine peptide aglycone and processes or interpolation hydrolytic enzyme.
China application CN200910067227.7 has introduced a kind of manufacture method, its process is cleaning, stripping and slicing, homogenate, deeply freeze, heated and boiled, separation supernatant, acid-base precipitation, alkali deposited, hot pressing, deeply freeze, ultrafiltration, detection, adjusting polypeptide and hypoxanthic ratio, ultrafiltration.Aseptic filtration, fill, sterilizing makes sarcosine peptide glycoside injection liquid, adds appropriate adjuvant aseptic filtration, fill, lyophilization, makes injection sarcosine peptide aglycone.Whole process is easy to be complete, ensures that product meets national standard, and has ensured inactivation of virus, and injection sterilizing F0 is greater than 8 simultaneously, and sterility assurance level is high.Its process need heated and boiled.
Chinese patent CN03104642.8 relates to a kind of sarcosine peptide aglycone powdery injection and preparation method thereof, it is characterized in that it is made up of polypeptide, hypoxanthine and excipient, the sarcosine peptide aglycone solution that adopts modern biological extraction technology to make, and use modern low-temperature freeze drying preparation technique to make aseptic freeze-dried injectable powder, injection after dissolving with injection water or transfusion when use, has good stability, long, safe advantage and the effect of storage time.Wherein the extraction temperature of sarcosine peptide aglycone reaches 80 DEG C～85 DEG C, keeps 5～8 minutes.
China application CN201210203244.0 relates to a kind of sarcosine peptide aglycone extract and compositions thereof containing drug regimen compositions such as polypeptide, aminoacid, nucleoside and nucleotide, its preparation comprises the steps: to get rabbit hearts and muscle, remove fat, homogenate, evacuation, pass into carbon dioxide, then evacuation, inflation repeatedly 2 times; Rabbit hearts saturated carbon dioxide and muscle slurry are joined respectively in liquid CO 2, stir, be cooled to-80 DEG C, keep 6～10h, be warming up to 40～50 DEG C, respectively get rabbit hearts and the muscle after freeze thawing at 1: 3 according to weight ratio, add respectively in water for injection, heart tissue solution is added to elastoser, muscular tissue solution is added to papain; Merge heart tissue solution and muscular tissue solution after enzymolysis, centrifugal, get supernatant, add the activated carbon of 0.3 times of weight, under room temperature, stir 30min, filtering with microporous membrane and get final product.Its process need utilizes carbon dioxide and hydrolytic enzyme.
In prior art scheme taking above-mentioned technical scheme as representative, very high-temperature and interpolation hydrolytic enzyme are adopted in order to obtain qualified sarcosine peptide aglycone extract, and higher temperature can destroy to some extent to effective ingredient, add hydrolytic enzyme technique comparatively complicated, cost is higher, and hydrolytic enzyme one's duty also unstable needs strictly control.The use dosage form of sarcosine peptide aglycone is injection, and the variation such as polypeptide, aminoacid isoreactivity material are easily oxidized in storage, condensation, polymerization owing to containing easily causes particulate matter significantly to increase and less stable.
In order to address the above problem, the inventor, at long-term large quantity research, obtains a kind of sarcosine peptide aglycone extracting method and high sarcosine peptide aglycone pharmaceutical composition of stability that higher temperature or hydrolytic enzyme and yield are high that do not need.
Summary of the invention
First the present invention provides a kind of sarcosine peptide aglycone extract, and described extract is prepared by the method comprising the steps:
1, get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without after color with water for injection, put in meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, adjust pH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature 1 hour.
2, homogenate is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, 3 times so repeatedly.
3, homogenate being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, homogenate being placed in to power is 1KW, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune PH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour.
4, by homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, by filtrate with 10000 molecular weight ultrafilter membrane ultrafiltration and get final product.
Sarcosine peptide aglycone extracting method of the present invention has been avoided high temperature or has been adopted hydrolytic enzyme hydrolysis, adopt freeze thawing and different temperatures, PH, mixing speed, power, ultrasonic field control under frequency gets off to extract sarcosine peptide aglycone, extracting method yield of the present invention is high, finish and close technique of the present invention and add potassium chloride by 2.5% of homogenate weight sarcosine peptide aglycone is extracted and has potentiation in the favorite outer discovery of research process, find through large quantity research, in the preparation method of sarcosine peptide aglycone extract of the present invention, each control main points is most important to the extraction effect of sarcosine peptide aglycone.
The present invention also provides a kind of stability high sarcosine peptide aglycone medicine composition injection, and described sarcosine peptide aglycone pharmaceutical composition consists of the following composition:
Sarcosine peptide aglycone solution 1000ml
Sorbitol: 1.5g
Thioglycerol: 0.2g
PLURONICS F87: 1.0g
Described sarcosine peptide aglycone solution: peptide concentration is: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml.
The preparation method of described sarcosine peptide aglycone medicine composition injection comprises the steps:
1, the preparation of sarcosine peptide aglycone solution:
1. get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without after color with water for injection, put in meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, adjust PH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature 1 hour.
2. homogenate is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, 3 times so repeatedly.
3. homogenate being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, homogenate being placed in to power is 1KW, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune PH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour.
4. by homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate obtained to sarcosine peptide aglycone extracting solution with 10000 molecular weight ultrafilter membrane ultrafiltration, measure polypeptide and hypoxanthine content, adjust solution concentration, inject water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml;
2, sorbitol, thioglycerol, PLURONICS F87 are added to sarcosine peptide aglycone solution successively, 28～32 DEG C of stirring and dissolving, 0.22 μ m membrane filtration, subpackage after the assay was approved, 121 DEG C of moist heat sterilizations 20 minutes, obtain sarcosine peptide aglycone medicine composition injection.
Under a large amount of development tests of the applicant, the stability that in sarcosine peptide aglycone medicine composition injection of the present invention, sorbitol, thioglycerol, PLURONICS F87 combining under use amount of the present invention used improving sarcosine peptide aglycone and the lower particulate matter of maintenance have played beyond thought effect (referring to table 1-2).
The different prescriptions of table 1 sarcosine peptide aglycone medicine composition injection
| Prescription | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| Sarcosine peptide aglycone solution (ml) | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 |
| Sorbitol (g) | — | 1.5 | — | — | 1.5 | 1.5 | — | 1.5 |
| Thioglycerol (g) | — | — | 0.2 | — | 0.2 | — | 0.2 | 0.2 |
| PLURONICS F87 (g) | — | — | — | 1.0 | — | 1.0 | 1.0 | 1.0 |
The injection liquid samples of above several formula preparations is placed 10 days under 60 DEG C of conditions, respectively at sampling calibrating in the 10th day, result and comparison in 0 day, the stability of investigation sample, the results are shown in Table 2:
The different prescription of table 2 sarcosine peptide aglycone medicine composition injection stability test
The present invention also provides a kind of stability high sarcosine peptide aglycone lyophilized injection of pharmaceutical composition, and described sarcosine peptide aglycone pharmaceutical composition consists of the following composition:
Sarcosine peptide aglycone solution 1000ml
Dextran 40: 40g
Thioglycerol: 0.2g
PLURONICS F87: 1.0g
HYDROXYPROPYL BETA-CYCLODEXTRIN: 0.5g
The preparation method of described sarcosine peptide aglycone lyophilized injection of pharmaceutical composition comprises the steps:
1, the preparation of sarcosine peptide aglycone solution:
1. get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without after color with water for injection, put in meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, adjust PH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature 1 hour.
2. homogenate is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, 3 times so repeatedly.
3. homogenate being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, homogenate being placed in to power is 1KW, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune PH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour.
4. by homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate obtained to sarcosine peptide aglycone extracting solution with 10000 molecular weight ultrafilter membrane ultrafiltration, measure polypeptide and hypoxanthine content, adjust solution concentration, inject water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml;
2, successively Dextran 40, thioglycerol, PLURONICS F87, HYDROXYPROPYL BETA-CYCLODEXTRIN are added in sarcosine peptide aglycone solution to 28～32 DEG C of stirring and dissolving in prescription ratio.
3, toward the active carbon that adds 0.10g/100ml in 2, stir 20 minutes, 0.22 μ m membrane filtration degerming, measures filtrate pH value, content, determines fill amount, the subpackage score dress liquid of partly jumping a queue.
4, lyophilizing:
1. pre-freeze: subpackage liquid is placed in advance and is cooled in-37 DEG C～-45 DEG C household freezers, keep 2～3 hours;
2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (1～2 DEG C/h) to-30 DEG C～-27 DEG C, keep 12 hours in this temperature.At the uniform velocity heat up (1～1.5 DEG C/h) to-20 DEG C～-15 DEG C, keep 6 hours in this temperature.
3. dry: to be at the uniform velocity warming up to 37 DEG C, dry 8 hours, to detect qualified rear packaging warehouse-in.
Under a large amount of development tests of the applicant, the stability that in sarcosine peptide aglycone lyophilized injection of pharmaceutical composition of the present invention, Dextran 40, thioglycerol, PLURONICS F87 and HYDROXYPROPYL BETA-CYCLODEXTRIN combining under use amount of the present invention used improving sarcosine peptide aglycone and the lower particulate matter of maintenance have played beyond thought effect (referring to table 3-4).
The different prescriptions of table 3 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition
| Prescription | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| Sarcosine peptide aglycone solution (ml) | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 | 1000 |
| Dextran 40 (g) | 40 | 40 | 40 | 40 | 40 | 40 | 40 | 40 |
| Thioglycerol (g) | — | — | 0.2 | — | 0.2 | — | 0.2 | 0.2 |
| PLURONICS F87 (g) | — | — | — | 1.0 | 1.0 | 1.0 | — | 1.0 |
| HYDROXYPROPYL BETA-CYCLODEXTRIN (g) | — | 0.5 | — | — | — | 0.5 | 0.5 | 0.5 |
The freeze-dried powder sample of above several formula preparations is placed 10 days under 60 DEG C of conditions, respectively at sampling calibrating in the 10th day, result and comparison in 0 day, the stability of investigation sample, the results are shown in Table 4:
The different prescription of table 4 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition stability test
detailed description of the invention
The preparation of embodiment 1 sarcosine peptide aglycone extract
Get cardiac muscle and muscle (weight ratio 1:1) 1000g altogether of healthy rabbit, remove connective tissue, rinse to without after color with water for injection, put in meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68:1 with water for injection, adjust PH to 5.7 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 24 DEG C, constant temperature 1 hour; Homogenate is thawed in 40 DEG C after freezing 12 hours at-30 DEG C, 3 times so repeatedly; It is 0.8KW that homogenate is placed in to power, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, homogenate being placed in to power is 1KW, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25 DEG C, with acetic acid tune PH to 5.2, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour.By homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate obtained to (sarcosine peptide aglycone extract polypeptide and hypoxanthic content: polypeptide 11.2g with 10000 molecular weight ultrafilter membrane ultrafiltration, hypoxanthine 1.65 g, detection method, polypeptide: Forint phenol method; Hypoxanthine: high performance liquid chromatography).
The preparation of embodiment 2 sarcosine peptide aglycone extracts
Get cardiac muscle and muscle (weight ratio 1:1) 1000g altogether of healthy rabbit, remove connective tissue, rinse to without after color with water for injection, put in meat grinder and rub, minced tissues is mixed by weight the ratio of 0.72:1 with water for injection, adjust pH to 6.0 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 27 DEG C, constant temperature 1 hour; Homogenate is thawed in 44 DEG C after freezing 12 hours at-32 DEG C, 3 times so repeatedly; It is 0.8KW that homogenate is placed in to power, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, homogenate being placed in to power is 1KW, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 28 DEG C, with acetic acid tune PH to 5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour.By homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate obtained to (sarcosine peptide aglycone extract polypeptide and hypoxanthic content: polypeptide 11.3g, hypoxanthine 1.67 is g) with 10000 molecular weight ultrafilter membrane ultrafiltration.
The preparation (5ml) of embodiment 3 sarcosine peptide aglycone medicine composition injections
Get sarcosine peptide aglycone extract prepared by embodiment 1, measure polypeptide and hypoxanthine content in solution, adjust solution concentration, inject water to 1000ml and make peptide concentration be: 1.74mg/ml, hypoxanthine concentration is: 0.25mg/ml; Sorbitol 1.5g, thioglycerol 0.2g, PLURONICS F87 1.0g are added to sarcosine peptide aglycone solution successively, 32 DEG C of stirring and dissolving, 0.22 μ m membrane filtration, subpackage after the assay was approved, 121 DEG C of moist heat sterilizations 20 minutes, obtain sarcosine peptide aglycone medicine composition injection.
The preparation (2ml) of embodiment 4 sarcosine peptide aglycone medicine composition injections
Get sarcosine peptide aglycone extract prepared by embodiment 2, measure polypeptide and hypoxanthine content in solution, adjust solution concentration, inject water to 1000ml and make peptide concentration be: 1.75mg/ml, hypoxanthine concentration is: 0.26mg/ml; Sorbitol 1.5g, thioglycerol 0.2g, PLURONICS F87 1.0g are added to sarcosine peptide aglycone solution successively, 30 DEG C of stirring and dissolving, 0.22 μ m membrane filtration, subpackage after the assay was approved, 121 DEG C of moist heat sterilizations 20 minutes, obtain sarcosine peptide aglycone medicine composition injection.
The preparation of embodiment 5 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition
Get sarcosine peptide aglycone extract prepared by embodiment 1, measure polypeptide and hypoxanthine content in solution, adjust solution concentration, inject water to 1000ml and make peptide concentration be: 1.75mg/ml, hypoxanthine concentration is: 0.25mg/ml; Successively Dextran 40 40g, thioglycerol 0.2g, PLURONICS F87 1.0g, HYDROXYPROPYL BETA-CYCLODEXTRIN 0.5g are added in sarcosine peptide aglycone solution to 32 DEG C of stirring and dissolving in prescription ratio; Add the active carbon of 0.10g/100ml, stir 20 minutes, 0.22 μ m membrane filtration degerming, measures filtrate pH value, content, determines fill amount, the subpackage score dress liquid of partly jumping a queue; Lyophilizing: 1. pre-freeze: subpackage liquid is placed in advance and is cooled in-37 DEG C of household freezers, keep 2 hours; 2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (1 DEG C/h) to-30 DEG C, keep 12 hours in this temperature.At the uniform velocity heat up (1 DEG C/h) to-20 DEG C, keep 6 hours in this temperature.3. dry: to be at the uniform velocity warming up to 37 DEG C, dry 8 hours, to detect qualified rear packaging warehouse-in.
The preparation of embodiment 6 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition
Get sarcosine peptide aglycone extract prepared by embodiment 2, measure polypeptide and hypoxanthine content in solution, adjust solution concentration, inject water to 1000ml and make peptide concentration be: 1.74mg/ml, hypoxanthine concentration is: 0.26mg/ml; Successively 40g Dextran 40,0.2g thioglycerol, 1.0g PLURONICS F87,0.5g HYDROXYPROPYL BETA-CYCLODEXTRIN are added in sarcosine peptide aglycone solution to 32 DEG C of stirring and dissolving in prescription ratio; Add the active carbon of 0.10g/100ml, stir 20 minutes, 0.22 μ m membrane filtration degerming, measures filtrate pH value, content, determines fill amount, the subpackage score dress liquid of partly jumping a queue; Lyophilizing: 1. pre-freeze: subpackage liquid is placed in advance and is cooled in-45 DEG C of household freezers, keep 3 hours; 2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (2 DEG C/h) to-27 DEG C, keep 12 hours in this temperature.At the uniform velocity heat up (1.5 DEG C/h) to-15 DEG C, keep 6 hours in this temperature.3. dry: to be at the uniform velocity warming up to 37 DEG C, dry 8 hours, to detect qualified rear packaging warehouse-in.
The invention provides following test example and comparing result:
Test example 1 sarcosine peptide aglycone extraction effect contrast
1, the sarcosine peptide aglycone extract that sample 1(is prepared by the embodiment of the present invention 1 method)
2, the sarcosine peptide aglycone extract that sample 2(is prepared by the embodiment of the present invention 2 methods)
3, sample 3(, by the embodiment of the present invention 1 method, does not add sarcosine peptide aglycone extract prepared by potassium chloride)
4, the sarcosine peptide aglycone extract that sample 4(is prepared by CN1522757A embodiment 1)
5, the sarcosine peptide aglycone extract that sample 5(is prepared by CN102727428A embodiment 1)
6, the sarcosine peptide aglycone extract that sample 6(is prepared by CN102697810A embodiment 1)
For parallel contrast, sample 1～6 uses with a collection of Carnis Leporis Cor Leporis raw material and all gets Rabbit Heart and muscle 1000g preparation altogether by weight 1:1, checks polypeptide and hypoxanthic yield, and result of the test is in table 5.
Table 5 sarcosine peptide aglycone extraction effect result of the test
Result of the test shows that the sarcosine peptide aglycone yield of the method for the invention is higher.
Test example 2 sarcosine peptide aglycone medicine composition injection study on the stability
1, the sarcosine peptide aglycone medicine composition injection that sample 1(is prepared by the embodiment of the present invention 3)
2, the sarcosine peptide glycoside injection liquid that sample 2(is prepared by CN1824294A embodiment 1)
3, the sarcosine peptide glycoside injection liquid that sample 3(is prepared by CN101584713A)
4, the sarcosine peptide glycoside injection liquid that sample 4(is prepared by CN102727428A)
5, the sarcosine peptide glycoside injection liquid that sample 5(is prepared by CN102266351A embodiment 1)
6, the sarcosine peptide glycoside injection liquid that sample 6(is prepared by CN102847138A embodiment 1)
Sample 1～6 is carried out to accelerated stability investigation (40 DEG C ± 2 DEG C, RH 75% ± 5%), the results are shown in Table 6:
Table 6 sarcosine peptide glycoside injection liquid accelerated test result
Above-mentioned result of the test shows, sarcosine peptide aglycone medicine composition injection good stability prepared by the present invention, and particulate matter is low to be compared and has obvious advantage with prior art.
Test example 3 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition study on the stability
1, the sarcosine peptide aglycone lyophilized injection of pharmaceutical composition that sample 1(is prepared by the embodiment of the present invention 5)
2, the sarcosine peptide aglycone freeze-dried powder that sample 2(is prepared by CN1522757A embodiment 1)
3, the sarcosine peptide aglycone freeze-dried powder that sample 3(is prepared by CN1939533A embodiment 3)
4, sample 4(commercially available sarcosine peptide aglycone freeze-dried powder)
Sample 1～4 is carried out to accelerated stability investigation (40 DEG C ± 2 DEG C, RH 75% ± 5%), the results are shown in Table 7:
Table 7 sarcosine peptide aglycone freeze-dried powder accelerated test result
Above-mentioned result of the test shows, sarcosine peptide aglycone lyophilized injection of pharmaceutical composition good stability prepared by the present invention, and particulate matter is low to be compared and has obvious advantage with prior art.
Claims (3)
1. the preparation method of sarcosine peptide aglycone extract, comprises the steps:
(1) get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without rubbing after color with water for injection, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, with acetic acid tune pH to 5.7～6.2, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature obtains homogenate I for 1 hour;
(2) homogenate I is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, so repeatedly 3 times homogenate II;
(3) homogenate II being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, being placed in power is 1KW again, frequency is under the ultrasonic field of 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune pH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stir after 1 hour, close ultrasonic field, stop stirring, leave standstill 1 hour and obtain homogenate III,
(4) by homogenate III with 4000r/min centrifugal 25 minutes, get supernatant, obtain filtrate with 0.22 μ m membrane filtration, filtrate obtained to sarcosine peptide aglycone extract with 10000 molecular weight ultrafilter membrane ultrafiltration.
2. the preparation method of sarcosine peptide aglycone medicine composition injection, described sarcosine peptide aglycone medicine composition injection consists of the following composition: 1000ml sarcosine peptide aglycone solution, 1.5g sorbitol, 0.2g thioglycerol and 1.0g PLURONICS F87; It is characterized in that comprising the steps:
(1) preparation of sarcosine peptide aglycone solution:
1. get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without rubbing after color with water for injection, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, with acetic acid tune pH to 5.7～6.2, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature obtains homogenate I for 1 hour;
2. homogenate I is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, so repeatedly 3 times homogenate II;
3. homogenate II being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, being placed in power is 1KW again, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune pH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour and obtain homogenate III;
4. by homogenate III with 4000r/min centrifugal 25 minutes, get supernatant, obtain filtrate with 0.22 μ m membrane filtration, filtrate obtained to sarcosine peptide aglycone extract with 10000 molecular weight ultrafilter membrane ultrafiltration;
5. the sarcosine peptide aglycone extract 4. step being obtained is adjusted solution concentration, injects water and makes peptide concentration be: 1.58～1.92mg/ml, and hypoxanthine concentration is: 0.22～0.28mg/ml obtains sarcosine peptide aglycone solution;
(2) sorbitol, thioglycerol, PLURONICS F87 are added to sarcosine peptide aglycone solution successively, 28～32 DEG C of stirring and dissolving, 0.22 μ m membrane filtration, subpackage after the assay was approved, 121 DEG C of moist heat sterilizations 20 minutes, obtain sarcosine peptide aglycone medicine composition injection.
3. the preparation method of sarcosine peptide aglycone lyophilized injection of pharmaceutical composition, described in
?sarcosine peptide aglycone lyophilized injection of pharmaceutical composition is made by 1000ml sarcosine peptide aglycone solution, 40g Dextran 40,0.2g thioglycerol, 1.0g PLURONICS F87 and the lyophilizing of 0.5g HYDROXYPROPYL BETA-CYCLODEXTRIN; It is characterized in that comprising the steps:
(1) preparation of sarcosine peptide aglycone solution:
1. get cardiac muscle and the muscle of healthy rabbit, remove connective tissue, rinse to without rubbing after color with water for injection, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, with acetic acid tune pH to 5.7～6.2, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 DEG C, constant temperature obtains homogenate I for 1 hour;
2. homogenate I is thawed in 42 ± 2 DEG C after freezing 12 hours at-30 ± 2 DEG C, so repeatedly 3 times homogenate II;
3. homogenate II being placed in to power is 0.8KW, frequency is under the ultrasonic field of 25KHz, be the potassium chloride of homogenate weight 2.5% toward adding weight in homogenate, keep temperature to be 20～25 DEG C and open stirring, mixing speed is 250 revs/min, under this ultrasonic field, stir 2 hours, after 2 hours, being placed in power is 1KW again, and under the ultrasonic field that frequency is 40KHz, keeping temperature is 25～28 DEG C, with acetic acid tune pH to 4.7～5.5, mixing speed is 150 revs/min, under this ultrasonic field, stirs after 1 hour, closes ultrasonic field, stop stirring, leave standstill 1 hour and obtain homogenate III;
4. by homogenate III with 4000r/min centrifugal 25 minutes, get supernatant, obtain filtrate with 0.22 μ m membrane filtration, filtrate obtained to sarcosine peptide aglycone extract with 10000 molecular weight ultrafilter membrane ultrafiltration;
5. the sarcosine peptide aglycone extract 4. step being obtained is adjusted solution concentration, injects water and makes peptide concentration be: 1.58～1.92mg/ml, and hypoxanthine concentration is: 0.22～0.28mg/ml obtains sarcosine peptide aglycone solution;
(2) successively Dextran 40, thioglycerol, PLURONICS F87, HYDROXYPROPYL BETA-CYCLODEXTRIN are added in sarcosine peptide aglycone solution, 28～32 DEG C of stirring and dissolving obtain solution;
(3) in the solution obtaining toward step (2), add the active carbon of 0.10g/100ml, stir 20 minutes, 0.22 μ m membrane filtration degerming, measures filtrate pH value, content, determines fill amount, the subpackage score dress liquid of partly jumping a queue;
(4) lyophilizing:
1. pre-freeze: subpackage liquid is placed in advance and is cooled in-37 DEG C～-45 DEG C household freezers, keep 2～3 hours;
2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, and the speed of 1～2 DEG C/h is at the uniform velocity warming up to-30 DEG C～-27 DEG C, keeps 12 hours in this temperature; At the uniform velocity the speed of 1～1.5 DEG C/h is warming up to-20 DEG C～-15 DEG C, keeps 6 hours in this temperature;
3. dry: be at the uniform velocity warming up to 37 DEG C, dry 8 hours, packaging warehouse-in.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310153538.1A CN103191153B (en) | 2013-04-28 | 2013-04-28 | Muscular amino acid and nucleoside extract and pharmaceutical composition thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310153538.1A CN103191153B (en) | 2013-04-28 | 2013-04-28 | Muscular amino acid and nucleoside extract and pharmaceutical composition thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103191153A CN103191153A (en) | 2013-07-10 |
| CN103191153B true CN103191153B (en) | 2014-06-04 |
Family
ID=48714089
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310153538.1A Active CN103191153B (en) | 2013-04-28 | 2013-04-28 | Muscular amino acid and nucleoside extract and pharmaceutical composition thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103191153B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110478363B (en) * | 2019-08-22 | 2022-09-06 | 北京大学 | New medicinal application of cIMP |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824294A (en) * | 2005-12-21 | 2006-08-30 | 白求恩医科大学制药厂 | Preparation technology of sarcosine peptide glycoside injection liquid |
| CN102727428A (en) * | 2011-04-12 | 2012-10-17 | 河南科伦药业有限公司 | Preparation method of muscular amino acids and nucleosides injection |
-
2013
- 2013-04-28 CN CN201310153538.1A patent/CN103191153B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824294A (en) * | 2005-12-21 | 2006-08-30 | 白求恩医科大学制药厂 | Preparation technology of sarcosine peptide glycoside injection liquid |
| CN102727428A (en) * | 2011-04-12 | 2012-10-17 | 河南科伦药业有限公司 | Preparation method of muscular amino acids and nucleosides injection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103191153A (en) | 2013-07-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2012097575A1 (en) | Method for preparing pharmaceutical composition for enhancing immunity and pharmaceutical composition prepared according to the method | |
| CN101433553B (en) | Method for preparing deproteinized extract of calf blood and freeze-dried powder thereof | |
| US20180193373A1 (en) | Method for Preparing Linseed Polysaccharide Having Antiviral Activity and Immunological Activity, and Use of the Linseed Polysaccharide | |
| CN105037577B (en) | Procoagulant blackberry seed polysaccharide, and extraction separation method and application thereof | |
| CN101020715B (en) | Process of extracting and preparing deer nerve growth factor (DEER NGF) | |
| CN110354141A (en) | A kind of preparation method of deer blood freeze-dried powder | |
| CN104877035A (en) | Preparation method of auricularia polysaccharide with hypoglycemic effect | |
| CN106632718B (en) | The preparation method of bletilla polysaccharide | |
| CN104367987A (en) | Astragalus mongholicus formulation for veterinary use and preparation method thereof | |
| CN112791103B (en) | Deer blood product and preparation method thereof | |
| CN103191153B (en) | Muscular amino acid and nucleoside extract and pharmaceutical composition thereof | |
| CN107510705A (en) | A kind of freeze drying protectant of leech crude extract and lyophilized preservation technique | |
| CN114989258B (en) | Application of plant extract composition in preparing product for treating constipation and reducing weight | |
| CN101054414B (en) | Method of extracting and preparing deer DGF | |
| WO2016082327A1 (en) | Processing method of cordyceps sinensis mycelium | |
| CN115160450A (en) | Rapid preparation method and application of Pholiota nameko polysaccharide | |
| CN101337070B (en) | Freeze-dried attenuated live vaccine for hepatitis A and its preparing process | |
| CN102274241B (en) | Method for extracting sheep embryo extract and medicinal composition comprising same | |
| CN101081865B (en) | Abstraction of pilose antler releasing somatomedin (DEER GHRF) and preparation method thereof | |
| CN106472934B (en) | Mushroom concentrated beverage for improving immunity and preparation method thereof | |
| CN103655490A (en) | Idarubicin hydrochloride pharmaceutical composition and preparation method thereof | |
| CN104688766B (en) | Polysaccharide sulphate for injection powder-injection pharmaceutical composition and preparation method | |
| CN110467685B (en) | Preparation and purification method and application of cedrela sinensis seed polysaccharide | |
| CN111732672B (en) | Okra polygalacturonic acid with uric acid reducing effect and preparation method and application thereof | |
| CN115399447B (en) | Turtle blood crystal and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: HUNAN WUZHOUTONG PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: HUBEI NEO-TREATION PHARMACEUTICAL CO., LTD. Effective date: 20150316 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 430014 WUHAN, HUBEI PROVINCE TO: 411200 XIANGTAN, HUNAN PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20150316 Address after: 411200 Fenghuang Road, easy Town, Xiangtan County, Hunan Patentee after: HUNAN WUZHOUTONG PHARMACEUTICAL CO., LTD. Address before: 430014, Zhenghe Plaza, 707 Zhongshan Avenue, Jiang'an District, Hubei, Wuhan, 1506 Patentee before: Hubei Neo-Treation Pharmaceutical Co., Ltd. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Muscular amino acid and nucleoside extract and pharmaceutical composition thereof Effective date of registration: 20200629 Granted publication date: 20140604 Pledgee: China Construction Bank Corporation Xiangtan County sub branch Pledgor: HUNAN WUZHOUTONG PHARMACEUTICAL Co.,Ltd. Registration number: Y2020980003555 |
|
| CP03 | Change of name, title or address |
Address after: No. 219, biquantan Road, Tianyi Economic Development Zone, Xiangtan City, Hunan Province Patentee after: Hunan wuzhoutong Pharmaceutical Co., Ltd Address before: 411200 Fenghuang Road, easy Town, Xiangtan County, Hunan Patentee before: HUNAN WUZHOUTONG PHARMACEUTICAL Co.,Ltd. |
|
| CP03 | Change of name, title or address |