CN103191153A - Muscular amino acid and nucleoside extract and pharmaceutical composition thereof - Google Patents
Muscular amino acid and nucleoside extract and pharmaceutical composition thereof Download PDFInfo
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- CN103191153A CN103191153A CN2013101535381A CN201310153538A CN103191153A CN 103191153 A CN103191153 A CN 103191153A CN 2013101535381 A CN2013101535381 A CN 2013101535381A CN 201310153538 A CN201310153538 A CN 201310153538A CN 103191153 A CN103191153 A CN 103191153A
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- homogenate
- sarcosine peptide
- peptide aglycone
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- injection
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- 239000000284 extract Substances 0.000 title claims abstract description 34
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 21
- 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
- 238000002347 injection Methods 0.000 claims abstract description 53
- 239000007924 injection Substances 0.000 claims abstract description 53
- 239000000243 solution Substances 0.000 claims abstract description 49
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 claims abstract description 48
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 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 24
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 20
- 239000000203 mixture Substances 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
- 229920001993 poloxamer 188 Polymers 0.000 claims abstract description 17
- 229940044519 poloxamer 188 Drugs 0.000 claims abstract description 17
- 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 16
- 241000283973 Oryctolagus cuniculus Species 0.000 claims abstract description 13
- 210000003205 muscle Anatomy 0.000 claims abstract description 13
- 210000004165 myocardium Anatomy 0.000 claims abstract description 10
- 239000001103 potassium chloride Substances 0.000 claims abstract description 10
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 10
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 10
- 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
- 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
- 239000000600 sorbitol Substances 0.000 claims abstract description 9
- 210000002808 connective tissue Anatomy 0.000 claims abstract description 8
- 239000012528 membrane Substances 0.000 claims abstract description 8
- 229940119744 dextran 40 Drugs 0.000 claims abstract description 7
- CTKXFMQHOOWWEB-UHFFFAOYSA-N Ethylene oxide/propylene oxide copolymer Chemical compound CCCOC(C)COCCO CTKXFMQHOOWWEB-UHFFFAOYSA-N 0.000 claims abstract 6
- 108010077895 Sarcosine Proteins 0.000 claims description 113
- 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 106
- 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 106
- 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 106
- 238000002360 preparation method Methods 0.000 claims description 44
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 42
- 238000003756 stirring Methods 0.000 claims description 31
- 238000012856 packing Methods 0.000 claims description 20
- 239000003814 drug Substances 0.000 claims description 18
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- 210000001519 tissue Anatomy 0.000 claims description 16
- 238000005374 membrane filtration Methods 0.000 claims description 15
- 238000002156 mixing Methods 0.000 claims description 15
- 239000008215 water for injection Substances 0.000 claims description 15
- 238000012546 transfer Methods 0.000 claims description 14
- 239000000706 filtrate Substances 0.000 claims description 13
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000011010 flushing procedure Methods 0.000 claims description 7
- 238000007710 freezing Methods 0.000 claims description 7
- 230000008014 freezing Effects 0.000 claims description 7
- 238000005360 mashing Methods 0.000 claims description 7
- 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
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 238000003556 assay Methods 0.000 claims description 4
- 238000004821 distillation 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
- 229960002920 sorbitol Drugs 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 2
- 125000003729 nucleotide group Chemical group 0.000 abstract description 2
- 239000008176 lyophilized powder Substances 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
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 12
- 238000012360 testing method Methods 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 7
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- 229940088598 enzyme Drugs 0.000 description 7
- 238000000605 extraction Methods 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
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 235000013372 meat Nutrition 0.000 description 5
- 239000013618 particulate matter Substances 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 3
- 239000001569 carbon dioxide Substances 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 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
- 206010003549 asthenia Diseases 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
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- 210000005003 heart tissue Anatomy 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
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- 208000011580 syndromic disease Diseases 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical class 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
- 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
- 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
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- 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
- 150000003838 adenosines Chemical class 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
- 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
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- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
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- 229920001983 poloxamer Polymers 0.000 description 1
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- 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 is to contain atrial natriuretic peptide, calcitonin-gene-related peptide and adenosine triphosphate nucleotide (nucleoside) classes such as (adenosines), the medicine of 20 several amino acids and metabolic intermediate isoreactivity material thereof by what healthy rabbits muscle and cardiac muscle extracted.The effect of the organism metabolism of promotion is arranged.Be used for amyotrophy, nervous edema, cerebrovascular accident paralysis, nerve asthenia syndrome etc.The ischemic heart, 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.The 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 handles or the interpolation hydrolytic enzyme.
China application CN200910067227.7 has introduced a kind of manufacture method, its process is cleaning, stripping and slicing, homogenate, freeze deeply, heated and boiled, separation supernatant, acid-base precipitation, alkali deposited, hot pressing, freeze deeply, ultrafiltration, detection, adjusting polypeptide and hypoxanthic ratio, ultrafiltration.Aseptic filtration, fill, sterilization makes sarcosine peptide glycoside injection liquid, adds an amount of adjuvant aseptic filtration, fill, lyophilization, makes injection sarcosine peptide aglycone.Whole process is easy to be complete, guarantees that product meets national standard, and has guaranteed inactivation of virus, and F0 is greater than 8 in injection sterilization simultaneously, aseptic assurance level height.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, with injection after injection water or the transfusion dissolving, have good stability, long, safe advantage and the effect of storage time during use.Wherein the extraction temperature of sarcosine peptide aglycone reaches 80 ℃～85 ℃, keeps 5～8 minutes.
China application CN201210203244.0 relates to a kind of sarcosine peptide aglycone extract and compositions thereof that contains 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, feed carbon dioxide, evacuation, inflation are 2 times repeatedly again; The rabbit hearts that carbon dioxide is saturated and muscle slurry join in the liquid CO 2 respectively, stir, be cooled to-80 ℃, keep 6～10h, be warming up to 40～50 ℃, respectively get rabbit hearts and muscle after the freeze thawing at 1: 3 according to weight ratio, add in the water for injection respectively, heart tissue solution is added elastoser, muscular tissue solution is added papain; Heart tissue solution and muscular tissue solution behind the merging enzymolysis, centrifugal, get supernatant, add the activated carbon of 0.3 times of weight, stir 30min under the room temperature, filtering with microporous membrane is namely.Its process need utilizes carbon dioxide and hydrolytic enzyme.
Be in the prior art scheme of representative with above-mentioned technical scheme, very high-temperature and interpolation hydrolytic enzyme have been adopted in order to obtain qualified sarcosine peptide aglycone extract, and higher temperature can destroy to some extent to effective ingredient, it is comparatively complicated to add hydrolytic enzyme technology, cost is higher, needs strictness to control and the hydrolytic enzyme one's duty is also unstable.The dosage form of using of sarcosine peptide aglycone is injection, and owing to contain polypeptide, aminoacid isoreactivity material variations such as oxidation, condensation, polymerization takes place easily in storage, causes particulate matter significantly to increase and less stable easily.
In order to address the above problem, the inventor obtains a kind of high sarcosine peptide aglycone extracting method of higher temperature or hydrolytic enzyme and yield and high sarcosine peptide aglycone pharmaceutical composition of stability of not needing at long-term big quantity research.
Summary of the invention
The present invention at first provides a kind of sarcosine peptide aglycone extract, and described extract is prepared by the method that comprises the steps:
1, gets cardiac muscle and the muscle of healthy rabbit, remove connective tissue, with water for injection flushing after do not have color, put in the meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, transfer pH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 ℃, constant temperature 1 hour.
2, homogenate was thawed 3 times so repeatedly in 42 ± 2 ℃ after under-30 ± 2 ℃ freezing 12 hours.
3, homogenate being placed power is 0.8KW, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20～25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, after 2 hours homogenate being placed power is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25～28 ℃, transfer PH to 4.7～5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, left standstill 1 hour.
4, with homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, with filtrate with 10000 molecular weight ultrafilter membrane ultrafiltration namely.
Sarcosine peptide aglycone extracting method of the present invention has been avoided high temperature or has been adopted the hydrolytic enzyme hydrolysis, adopt the ultrasonic field under freeze thawing and different temperatures, PH, mixing speed, power, the frequency to control the extraction sarcosine peptide aglycone, extracting method yield height of the present invention, unexpectedly in research process found that adding potassium chloride in conjunction with technology of the present invention by 2.5% of homogenate weight extracts sarcosine peptide aglycone potentiation is arranged, through discovering in a large number, each control main points is most important to the extraction effect of sarcosine peptide aglycone in the sarcosine peptide aglycone preparation method of extract of the present invention.
The present invention also provides a kind of stability high sarcosine peptide aglycone medicine composition injection, and described sarcosine peptide aglycone pharmaceutical composition is grouped into by following one-tenth:
Sarcosine peptide aglycone solution 1000ml
Sorbitol: 1.5g
Thioglycerol: 0.2g
Poloxamer 188:1.0g
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, with water for injection flushing after do not have color, put in the meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, transfer PH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 ℃, constant temperature 1 hour.
2. homogenate was thawed 3 times so repeatedly in 42 ± 2 ℃ after under-30 ± 2 ℃ freezing 12 hours.
3. homogenate being placed power is 0.8KW, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20～25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, after 2 hours homogenate being placed power is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25～28 ℃, transfer PH to 4.7～5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, left standstill 1 hour.
4. with homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate is namely got the sarcosine peptide aglycone extracting solution with 10000 molecular weight ultrafilter membrane ultrafiltration, measure polypeptide and hypoxanthine content, adjust solution concentration, add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml;
2, sorbitol, thioglycerol, poloxamer 188 are added sarcosine peptide aglycone solution successively, 28～32 ℃ of stirring and dissolving, 0.22 μ m membrane filtration, packing after the assay was approved, 121 ℃ of moist heat sterilizations 20 minutes namely get the sarcosine peptide aglycone medicine composition injection.
Under a large amount of development tests of the applicant, the use of uniting under use amount of the present invention of sorbitol, thioglycerol, poloxamer 188 has been played beyond thought effect (referring to table 1-2) to stability and the lower particulate matter of maintenance that improves sarcosine peptide aglycone in the sarcosine peptide aglycone medicine composition injection of the present invention.
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 |
| Poloxamer 188 (g) | — | — | — | 1.0 | — | 1.0 | 1.0 | 1.0 |
The injection liquid samples of above several prescription preparations was placed 10 days under 60 ℃ of conditions, and respectively at sampling calibrating in the 10th day, stability of sample was investigated in result and comparison in 0 day, 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 is grouped into by following one-tenth:
Sarcosine peptide aglycone solution 1000ml
Dextran 40: 40g
Thioglycerol: 0.2g
Poloxamer 188:1.0g
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, with water for injection flushing after do not have color, put in the meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, transfer PH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 ℃, constant temperature 1 hour.
2. homogenate was thawed 3 times so repeatedly in 42 ± 2 ℃ after under-30 ± 2 ℃ freezing 12 hours.
3. homogenate being placed power is 0.8KW, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20～25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, after 2 hours homogenate being placed power is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25～28 ℃, transfer PH to 4.7～5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, left standstill 1 hour.
4. with homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate is namely got the sarcosine peptide aglycone extracting solution with 10000 molecular weight ultrafilter membrane ultrafiltration, measure polypeptide and hypoxanthine content, adjust solution concentration, add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml;
2, successively Dextran 40, thioglycerol, poloxamer 188, HYDROXYPROPYL BETA-CYCLODEXTRIN are added in the sarcosine peptide aglycone solution 28～32 ℃ of stirring and dissolving in the prescription ratio.
3, add the active carbon of 0.10g/100ml in 2, stirred 20 minutes, 0.22 μ m membrane filtration degerming, mensuration filtrate pH value, content are determined the fill amount, packing false add plug gets packing liquid.
4, lyophilizing:
1. pre-freeze: packing liquid placed in advance be cooled in-37 ℃～-45 ℃ household freezers, kept 2～3 hours;
2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (1～2 ℃/h) to-30 ℃～-27 ℃, kept 12 hours in this temperature.At the uniform velocity heat up (1～1.5 ℃/h) to-20 ℃～-15 ℃, kept 6 hours in this temperature.
3. dry: as at the uniform velocity to be warming up to 37 ℃, dry 8 hours, to detect qualified back packing warehouse-in.
Under a large amount of development tests of the applicant, the use of uniting under use amount of the present invention of Dextran 40, thioglycerol, poloxamer 188 and HYDROXYPROPYL BETA-CYCLODEXTRIN has been played beyond thought effect (referring to table 3-4) to stability and the lower particulate matter of maintenance that improves sarcosine peptide aglycone in the sarcosine peptide aglycone lyophilized injection of pharmaceutical composition of the present invention.
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 |
| Poloxamer 188 (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 prescription preparations was placed 10 days under 60 ℃ of conditions, and respectively at sampling calibrating in the 10th day, stability of sample was investigated in result and comparison in 0 day, the results are shown in Table 4:
The different prescription of table 4 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition stability test
The specific embodiment
The preparation of embodiment 1 sarcosine peptide aglycone extract
Cardiac muscle and the muscle (weight ratio 1:1) of getting healthy rabbit are total to 1000g, remove connective tissue, with water for injection flushing after do not have color, put in the meat grinder and rub, minced tissues is mixed by weight the ratio of 0.68:1 with water for injection, transfer PH to 5.7 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 24 ℃, constant temperature 1 hour; Homogenate was thawed 3 times so repeatedly in 40 ℃ after under-30 ℃ freezing 12 hours; It is 0.8KW that homogenate is placed power, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, after 2 hours homogenate being placed power is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25 ℃, transfer PH to 5.2 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, left standstill 1 hour.With homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate is namely got (sarcosine peptide aglycone extract polypeptide and Determination of Hypoxanthine: 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
Cardiac muscle and the muscle (weight ratio 1:1) of getting healthy rabbit are total to 1000g, remove connective tissue, with water for injection flushing after do not have color, put in the meat grinder and rub, minced tissues is mixed by weight the ratio of 0.72:1 with water for injection, transfer pH to 6.0 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 27 ℃, constant temperature 1 hour; Homogenate was thawed 3 times so repeatedly in 44 ℃ after under-32 ℃ freezing 12 hours; It is 0.8KW that homogenate is placed power, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, after 2 hours homogenate being placed power is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 28 ℃, transfer PH to 5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, left standstill 1 hour.With homogenate with 4000r/min centrifugal 25 minutes, get supernatant, with 0.22 μ m membrane filtration, filtrate is namely got (sarcosine peptide aglycone extract polypeptide and Determination of Hypoxanthine: polypeptide 11.3g, hypoxanthine 1.67 g) with 10000 molecular weight ultrafilter membrane ultrafiltration.
The preparation (5ml) of embodiment 3 sarcosine peptide aglycone medicine composition injections
Get the sarcosine peptide aglycone extract of embodiment 1 preparation, measure polypeptide and hypoxanthine content in the solution, adjust solution concentration, add the injection water and make peptide concentration be to 1000ml: 1.74mg/ml, hypoxanthine concentration is: 0.25mg/ml; Sorbitol 1.5g, thioglycerol 0.2g, poloxamer 188 1.0g are added sarcosine peptide aglycone solution successively, 32 ℃ of stirring and dissolving, 0.22 μ m membrane filtration, packing after the assay was approved, 121 ℃ of moist heat sterilizations 20 minutes namely get the sarcosine peptide aglycone medicine composition injection.
The preparation (2ml) of embodiment 4 sarcosine peptide aglycone medicine composition injections
Get the sarcosine peptide aglycone extract of embodiment 2 preparation, measure polypeptide and hypoxanthine content in the solution, adjust solution concentration, add the injection water and make peptide concentration be to 1000ml: 1.75mg/ml, hypoxanthine concentration is: 0.26mg/ml; Sorbitol 1.5g, thioglycerol 0.2g, poloxamer 188 1.0g are added sarcosine peptide aglycone solution successively, 30 ℃ of stirring and dissolving, 0.22 μ m membrane filtration, packing after the assay was approved, 121 ℃ of moist heat sterilizations 20 minutes namely get the sarcosine peptide aglycone medicine composition injection.
The preparation of embodiment 5 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition
Get the sarcosine peptide aglycone extract of embodiment 1 preparation, measure polypeptide and hypoxanthine content in the solution, adjust solution concentration, add the injection water and make peptide concentration be to 1000ml: 1.75mg/ml, hypoxanthine concentration is: 0.25mg/ml; Successively Dextran 40 40g, thioglycerol 0.2g, poloxamer 188 1.0g, HYDROXYPROPYL BETA-CYCLODEXTRIN 0.5g are added in the sarcosine peptide aglycone solution 32 ℃ of stirring and dissolving in the prescription ratio; Add the active carbon of 0.10g/100ml, stirred 20 minutes, filtrate pH value, content are measured in 0.22 μ m membrane filtration degerming, determine the fill amount, and packing false add plug gets packing liquid; Lyophilizing: 1. pre-freeze: packing liquid placed in advance be cooled in-37 ℃ of household freezers, kept 2 hours; 2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (1 ℃/h) to-30 ℃, kept 12 hours in this temperature.At the uniform velocity heat up (1 ℃/h) to-20 ℃, kept 6 hours in this temperature.3. dry: as at the uniform velocity to be warming up to 37 ℃, dry 8 hours, to detect qualified back packing warehouse-in.
The preparation of embodiment 6 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition
Get the sarcosine peptide aglycone extract of embodiment 2 preparation, measure polypeptide and hypoxanthine content in the solution, adjust solution concentration, add the injection water and make peptide concentration be to 1000ml: 1.74mg/ml, hypoxanthine concentration is: 0.26mg/ml; Successively 40g Dextran 40,0.2g thioglycerol, 1.0g poloxamer 188,0.5g HYDROXYPROPYL BETA-CYCLODEXTRIN are added in the sarcosine peptide aglycone solution 32 ℃ of stirring and dissolving in the prescription ratio; Add the active carbon of 0.10g/100ml, stirred 20 minutes, filtrate pH value, content are measured in 0.22 μ m membrane filtration degerming, determine the fill amount, and packing false add plug gets packing liquid; Lyophilizing: 1. pre-freeze: packing liquid placed in advance be cooled in-45 ℃ of household freezers, kept 3 hours; 2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, at the uniform velocity heats up (2 ℃/h) to-27 ℃, kept 12 hours in this temperature.At the uniform velocity heat up (1.5 ℃/h) to-15 ℃, kept 6 hours in this temperature.3. dry: as at the uniform velocity to be warming up to 37 ℃, dry 8 hours, to detect qualified back packing warehouse-in.
The invention provides following test example and comparing result:
The contrast of test example 1 sarcosine peptide aglycone extraction effect
1, sample 1(is by the sarcosine peptide aglycone extract of the embodiment of the invention 1 method preparation)
2, sample 2(is by the sarcosine peptide aglycone extract of the embodiment of the invention 2 methods preparation)
3, sample 3(does not add the sarcosine peptide aglycone extract of potassium chloride preparation by the embodiment of the invention 1 method)
4, sample 4(presses the sarcosine peptide aglycone extract of CN1522757A embodiment 1 preparation)
5, sample 5(presses the sarcosine peptide aglycone extract of CN102727428A embodiment 1 preparation)
6, sample 6(presses the sarcosine peptide aglycone extract of CN102697810A embodiment 1 preparation)
For parallel contrast, sample 1～6 uses with a collection of Carnis Leporis Cor Leporis raw material and all gets tame rabbit heart and muscle 1000g preparation altogether by weight 1:1, checks polypeptide and hypoxanthic yield, and result of the test sees 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, sample 1(is by the sarcosine peptide aglycone medicine composition injection of the embodiment of the invention 3 preparations)
2, sample 2(presses the sarcosine peptide glycoside injection liquid of CN1824294A embodiment 1 preparation)
3, sample 3(presses the sarcosine peptide glycoside injection liquid of CN101584713A preparation)
4, sample 4(presses the sarcosine peptide glycoside injection liquid of CN102727428A preparation)
5, sample 5(presses the sarcosine peptide glycoside injection liquid of CN102266351A embodiment 1 preparation)
6, sample 6(presses the sarcosine peptide glycoside injection liquid of CN102847138A embodiment 1 preparation)
Sample 1～6 is carried out accelerated stability investigates (40 ℃ ± 2 ℃, 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, the sarcosine peptide aglycone medicine composition injection good stability of the present invention's preparation, and low the comparing with prior art of particulate matter has remarkable advantages.
Test example 3 sarcosine peptide aglycone lyophilized injection of pharmaceutical composition study on the stability
1, sample 1(is by the sarcosine peptide aglycone lyophilized injection of pharmaceutical composition of the embodiment of the invention 5 preparations)
2, sample 2(presses the sarcosine peptide aglycone freeze-dried powder of CN1522757A embodiment 1 preparation)
3, sample 3(presses the sarcosine peptide aglycone freeze-dried powder of CN1939533A embodiment 3 preparations)
4, sample 4(commercially available the sarcosine peptide aglycone freeze-dried powder)
Sample 1～4 is carried out accelerated stability investigates (40 ℃ ± 2 ℃, 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, the sarcosine peptide aglycone lyophilized injection of pharmaceutical composition good stability of the present invention's preparation, and low the comparing with prior art of particulate matter has remarkable advantages.
Claims (6)
1. sarcosine peptide aglycone extract, by the method preparation that comprises the steps:
(1) gets cardiac muscle and the muscle of healthy rabbit, remove connective tissue, do not rub after having color with the water for injection flushing, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, transfer pH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 ℃, constant temperature got the homogenate I in 1 hour;
(2) the homogenate I is thawed in 42 ± 2 ℃ after freezing 12 hours down at-30 ± 2 ℃, so repeatedly 3 times the homogenate II;
(3) the homogenate II being placed power is 0.8KW, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20～25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, placing power after 2 hours again is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25～28 ℃, transfer pH to 4.7～5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, leave standstill and got the homogenate III in 1 hour;
(4) with homogenate III with 4000r/min centrifugal 25 minutes, get supernatant, get filtrate with 0.22 μ m membrane filtration, filtrate is namely got the sarcosine peptide aglycone extract with 10000 molecular weight ultrafilter membrane ultrafiltration.
2. the sarcosine peptide aglycone preparation method of extract comprises the steps:
(1) gets cardiac muscle and the muscle of healthy rabbit, remove connective tissue, do not rub after having color with the water for injection flushing, minced tissues is mixed by weight the ratio of 0.68～0.72:1 with water for injection, transfer pH to 5.7～6.2 with acetic acid, use high-speed tissue mashing machine's homogenate, homogenized temperature is adjusted to 22～27 ℃, constant temperature got the homogenate I in 1 hour;
(2) the homogenate I is thawed in 42 ± 2 ℃ after freezing 12 hours down at-30 ± 2 ℃, so repeatedly 3 times the homogenate II;
(3) the homogenate II being placed power is 0.8KW, frequency is under the ultrasonic field of 25KHz, adding weight in the homogenate is the potassium chloride of homogenate weight 2.5%, keep temperature to be 20～25 ℃ and open stirring, mixing speed is 250 rev/mins, under this ultrasonic field, stirred 2 hours, placing power after 2 hours again is 1KW, and frequency is under the ultrasonic field of 40KHz, and keeping temperature is 25～28 ℃, transfer pH to 4.7～5.5 with acetic acid, mixing speed is 150 rev/mins, after stirring 1 hour under this ultrasonic field, closes ultrasonic field, stop to stir, leave standstill and got the homogenate III in 1 hour;
(4) with homogenate III with 4000r/min centrifugal 25 minutes, get supernatant, get filtrate with 0.22 μ m membrane filtration, filtrate is namely got the sarcosine peptide aglycone extract with 10000 molecular weight ultrafilter membrane ultrafiltration.
3. sarcosine peptide aglycone medicine composition injection, described sarcosine peptide aglycone pharmaceutical composition is grouped into by following one-tenth: 1000ml sarcosine peptide aglycone solution, 1.5g sorbitol, 0.2g thioglycerol and 1.0g poloxamer 188; Described sarcosine peptide aglycone solution is made by purgation: the sarcosine peptide aglycone extract that claim 1 obtains is adjusted solution concentration, and add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml.
4. the preparation method of the described sarcosine peptide aglycone medicine composition injection of claim 3 comprises the steps:
(1) preparation of sarcosine peptide aglycone solution:
The sarcosine peptide aglycone extract that claim 1 obtains is adjusted solution concentration, add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml gets sarcosine peptide aglycone solution;
(2) sorbitol, thioglycerol, poloxamer 188 are added sarcosine peptide aglycone solution successively, 28～32 ℃ of stirring and dissolving, 0.22 μ m membrane filtration, packing after the assay was approved, 121 ℃ of moist heat sterilizations 20 minutes namely get the sarcosine peptide aglycone medicine composition injection.
5. the sarcosine peptide aglycone lyophilized injection of pharmaceutical composition is made by 1000ml sarcosine peptide aglycone solution, 40g Dextran 40,0.2g thioglycerol, 1.0g poloxamer 188 and the lyophilizing of 0.5g HYDROXYPROPYL BETA-CYCLODEXTRIN; Described sarcosine peptide aglycone solution is made by purgation: the sarcosine peptide aglycone extract that claim 1 obtains is adjusted solution concentration, and add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml.
6. the preparation method of the described sarcosine peptide aglycone lyophilized injection of pharmaceutical composition of claim 5 comprises the steps:
(1) preparation of sarcosine peptide aglycone solution:
The sarcosine peptide aglycone extract that claim 1 obtains is adjusted solution concentration, add the injection water and make peptide concentration be: 1.58～1.92mg/ml, hypoxanthine concentration is: 0.22～0.28mg/ml gets sarcosine peptide aglycone solution;
(2) successively Dextran 40, thioglycerol, poloxamer 188, HYDROXYPROPYL BETA-CYCLODEXTRIN are added in the sarcosine peptide aglycone solution, 28～32 ℃ of stirring and dissolving get solution;
(3) add the active carbon of 0.10g/100ml in the solution that obtains toward step (2), stirred 20 minutes, 0.22 μ m membrane filtration degerming, mensuration filtrate pH value, content are determined the fill amount, packing false add plug gets packing liquid;
(4) lyophilizing:
1. pre-freeze: packing liquid placed in advance be cooled in-37 ℃～-45 ℃ household freezers, kept 2～3 hours;
2. distillation: open vacuum equipment, adjusting vacuum is 13Pa, and the speed of 1～2 ℃/h at the uniform velocity is warming up to-30 ℃～-27 ℃, keeps 12 hours in this temperature; At the uniform velocity the speed of 1～1.5 ℃/h is warming up to-20 ℃～-15 ℃, keeps 6 hours in this temperature;
3. dry: at the uniform velocity be warming up to 37 ℃, dry 8 hours, the packing warehouse-in.
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| 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 |
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| 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 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110478363A (en) * | 2019-08-22 | 2019-11-22 | 北京大学 | The new medicine use of cIMP |
| CN110478363B (en) * | 2019-08-22 | 2022-09-06 | 北京大学 | New medicinal application of cIMP |
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