CN1259925C - Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use - Google Patents
Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use Download PDFInfo
- Publication number
- CN1259925C CN1259925C CN 200410058086 CN200410058086A CN1259925C CN 1259925 C CN1259925 C CN 1259925C CN 200410058086 CN200410058086 CN 200410058086 CN 200410058086 A CN200410058086 A CN 200410058086A CN 1259925 C CN1259925 C CN 1259925C
- Authority
- CN
- China
- Prior art keywords
- coenzyme
- medicine
- complex
- yeast
- preparation
- 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.)
- Expired - Lifetime
Links
- 239000003814 drug Substances 0.000 title claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 title description 4
- 150000001875 compounds Chemical class 0.000 title description 3
- 239000005515 coenzyme Substances 0.000 claims abstract description 55
- 210000004027 cell Anatomy 0.000 claims abstract description 19
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 19
- 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 claims abstract description 14
- 238000002360 preparation method Methods 0.000 claims abstract description 13
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 claims abstract description 12
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 claims abstract description 12
- 229950006790 adenosine phosphate Drugs 0.000 claims abstract description 12
- 235000019162 flavin adenine dinucleotide Nutrition 0.000 claims abstract description 12
- 239000011714 flavin adenine dinucleotide Substances 0.000 claims abstract description 12
- VWWQXMAJTJZDQX-UYBVJOGSSA-N flavin adenine dinucleotide Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]([C@H](O)[C@@H]1O)O[C@@H]1CO[P@](O)(=O)O[P@@](O)(=O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C2=NC(=O)NC(=O)C2=NC2=C1C=C(C)C(C)=C2 VWWQXMAJTJZDQX-UYBVJOGSSA-N 0.000 claims abstract description 12
- 239000011768 flavin mononucleotide Substances 0.000 claims abstract description 12
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 claims abstract description 12
- 229940013640 flavin mononucleotide Drugs 0.000 claims abstract description 12
- FVTCRASFADXXNN-UHFFFAOYSA-N flavin mononucleotide Natural products OP(=O)(O)OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229940093632 flavin-adenine dinucleotide Drugs 0.000 claims abstract description 12
- 229950006238 nadide Drugs 0.000 claims abstract description 12
- 235000019231 riboflavin-5'-phosphate Nutrition 0.000 claims abstract description 12
- 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 claims abstract description 10
- RGJOEKWQDUBAIZ-IBOSZNHHSA-N CoASH Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCS)O[C@H]1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-IBOSZNHHSA-N 0.000 claims abstract description 10
- MEFKEPWMEQBLKI-AIRLBKTGSA-N S-adenosyl-L-methioninate Chemical compound O[C@@H]1[C@H](O)[C@@H](C[S+](CC[C@H](N)C([O-])=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 MEFKEPWMEQBLKI-AIRLBKTGSA-N 0.000 claims abstract description 10
- RGJOEKWQDUBAIZ-UHFFFAOYSA-N coenzime A Natural products OC1C(OP(O)(O)=O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000005516 coenzyme A Substances 0.000 claims abstract description 10
- 229940093530 coenzyme a Drugs 0.000 claims abstract description 10
- KDTSHFARGAKYJN-UHFFFAOYSA-N dephosphocoenzyme A Natural products OC1C(O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 KDTSHFARGAKYJN-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229960003180 glutathione Drugs 0.000 claims abstract description 10
- 238000011282 treatment Methods 0.000 claims abstract description 5
- 238000002512 chemotherapy Methods 0.000 claims abstract description 4
- UDMBCSSLTHHNCD-UHFFFAOYSA-N Coenzym Q(11) Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(O)=O)C(O)C1O UDMBCSSLTHHNCD-UHFFFAOYSA-N 0.000 claims abstract description 3
- 108010024636 Glutathione Proteins 0.000 claims abstract description 3
- 229960001570 ademetionine Drugs 0.000 claims abstract description 3
- 229960001456 adenosine triphosphate Drugs 0.000 claims abstract description 3
- 210000005229 liver cell Anatomy 0.000 claims abstract 2
- 230000002107 myocardial effect Effects 0.000 claims abstract 2
- 239000007788 liquid Substances 0.000 claims description 14
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 13
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 13
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- 238000001556 precipitation Methods 0.000 claims description 11
- 210000005253 yeast cell Anatomy 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical class CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 7
- 238000012546 transfer Methods 0.000 claims description 7
- 238000000108 ultra-filtration Methods 0.000 claims description 7
- 229910021529 ammonia Inorganic materials 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 6
- 238000001179 sorption measurement Methods 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 206010002660 Anoxia Diseases 0.000 claims description 5
- 241000976983 Anoxia Species 0.000 claims description 5
- 206010021143 Hypoxia Diseases 0.000 claims description 5
- 230000007953 anoxia Effects 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 5
- 230000006378 damage Effects 0.000 claims description 5
- 238000001728 nano-filtration Methods 0.000 claims description 5
- 239000008363 phosphate buffer Substances 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 4
- 238000001223 reverse osmosis Methods 0.000 claims description 4
- 206010003211 Arteriosclerosis coronary artery Diseases 0.000 claims description 3
- 206010008909 Chronic Hepatitis Diseases 0.000 claims description 3
- 244000168141 Geotrichum candidum Species 0.000 claims description 3
- 235000017388 Geotrichum candidum Nutrition 0.000 claims description 3
- 206010021245 Idiopathic thrombocytopenic purpura Diseases 0.000 claims description 3
- 206010030302 Oliguria Diseases 0.000 claims description 3
- 208000031981 Thrombocytopenic Idiopathic Purpura Diseases 0.000 claims description 3
- 230000001154 acute effect Effects 0.000 claims description 3
- 230000006907 apoptotic process Effects 0.000 claims description 3
- 201000003710 autoimmune thrombocytopenic purpura Diseases 0.000 claims description 3
- 230000001684 chronic effect Effects 0.000 claims description 3
- 208000029078 coronary artery disease Diseases 0.000 claims description 3
- 208000026758 coronary atherosclerosis Diseases 0.000 claims description 3
- 238000013467 fragmentation Methods 0.000 claims description 3
- 238000006062 fragmentation reaction Methods 0.000 claims description 3
- 208000006454 hepatitis Diseases 0.000 claims description 3
- 230000003907 kidney function Effects 0.000 claims description 3
- 208000010125 myocardial infarction Diseases 0.000 claims description 3
- 206010043554 thrombocytopenia Diseases 0.000 claims description 3
- 208000002381 Brain Hypoxia Diseases 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 2
- 230000001476 alcoholic effect Effects 0.000 claims description 2
- 238000005349 anion exchange Methods 0.000 claims description 2
- 210000002421 cell wall Anatomy 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 210000004907 gland Anatomy 0.000 claims description 2
- 230000035987 intoxication Effects 0.000 claims description 2
- 231100000566 intoxication Toxicity 0.000 claims description 2
- 238000004255 ion exchange chromatography Methods 0.000 claims description 2
- 238000012423 maintenance Methods 0.000 claims description 2
- 208000031225 myocardial ischemia Diseases 0.000 claims description 2
- 239000000575 pesticide Substances 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims 1
- 102100033925 GS homeobox 1 Human genes 0.000 claims 1
- 101001068303 Homo sapiens GS homeobox 1 Proteins 0.000 claims 1
- 238000009098 adjuvant therapy Methods 0.000 claims 1
- 230000005779 cell damage Effects 0.000 abstract description 3
- 208000037887 cell injury Diseases 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 3
- 210000004185 liver Anatomy 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 2
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 abstract 2
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 abstract 2
- 206010067125 Liver injury Diseases 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 231100000234 hepatic damage Toxicity 0.000 abstract 1
- 230000008818 liver damage Effects 0.000 abstract 1
- 230000004060 metabolic process Effects 0.000 abstract 1
- 238000011084 recovery Methods 0.000 description 18
- 241000700159 Rattus Species 0.000 description 9
- 238000002680 cardiopulmonary resuscitation Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 102000019197 Superoxide Dismutase Human genes 0.000 description 6
- 108010012715 Superoxide dismutase Proteins 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 4
- 238000005342 ion exchange Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 235000015170 shellfish Nutrition 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 231100000753 hepatic injury Toxicity 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 101150064015 FAS gene Proteins 0.000 description 2
- 208000010496 Heart Arrest Diseases 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000008351 acetate buffer Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000006837 decompression Effects 0.000 description 2
- 229940126534 drug product Drugs 0.000 description 2
- 235000003969 glutathione Nutrition 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 210000003701 histiocyte Anatomy 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000000849 parathyroid Effects 0.000 description 2
- 239000000825 pharmaceutical preparation Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- 208000037157 Azotemia Diseases 0.000 description 1
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 description 1
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 1
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- 206010068676 Pneumoretroperitoneum Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 101100537547 Rattus norvegicus Fas gene Proteins 0.000 description 1
- 208000005727 Retropneumoperitoneum Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 102000011923 Thyrotropin Human genes 0.000 description 1
- 108010061174 Thyrotropin Proteins 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000010247 heart contraction Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 230000008558 metabolic pathway by substance Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- AXOIZCJOOAYSMI-UHFFFAOYSA-N succinylcholine Chemical compound C[N+](C)(C)CCOC(=O)CCC(=O)OCC[N+](C)(C)C AXOIZCJOOAYSMI-UHFFFAOYSA-N 0.000 description 1
- 229940032712 succinylcholine Drugs 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 229960000874 thyrotropin Drugs 0.000 description 1
- 230000001748 thyrotropin Effects 0.000 description 1
- 229940034208 thyroxine Drugs 0.000 description 1
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 description 1
- 208000009852 uremia Diseases 0.000 description 1
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a preparation method of a complex coenzyme medicine, a complex coenzyme medicine thereof and clinical applications of the complex coenzyme medicine, which belongs to the technical field of medicines. The coenzyme components of the complex coenzyme medicine of the present invention comprise coenzyme A(CoA), coenzyme I(NAD), glutathione (GSH) and adenosine triphosphate (ATP); the present invention is characterized in that the coenzyme components also comprise flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD) adenosine diphosphate (ADP), adenosine phosphate (AMP) and ademetionine (SAM). By changing the existing producing technology of the complex coenzyme, the present invention increases the coenzyme substances to nine kinds from four kinds in the original complex coenzyme medicine product. The complex coenzyme can protect various cells such as liver cells, myocardial cells, etc., can maintain or restore the normal metabolism of the complex coenzyme and can be used for the clinical treatment for restoration of operations on the heart and the liver, the restoration of liver damage, the restoration of cell damage caused by rays or chemotherapy, etc.
Description
Technical field
The invention belongs to medical technical field, relate to a kind of preparation method and a kind of NEW TYPE OF COMPOSITE coenzyme medicine and clinical new purposes of new Coenzyme Complex medicine especially.
Background technology
Develop a kind of injection Coenzyme Complex lyophilized injectable powder the seventies, it is a kind of extraction from yeast (beer yeast, edible yeast and geotrichum candidum), contains coenzyme A (CoA), nadide (NAD), the natural coenzyme compound formulation of ATP and glutathion.Be used for the treatment of acute, chronic hepatitis, idiopathic thrombocytopenic purpura, thrombocytopenia for a long time, coronary atherosclerosis, chronic arterits, myocardial infarction, the weak oliguria that causes of renal function, uremia etc. be can be used as auxiliary therapeutic agent.
Yeast is a unicellular eukaryote, contains the necessary various coenzyme of biological substance metabolism such as sugar, fat, protein, nucleic acid (synthetic and decomposition) in its cell.But because original production process has been used the heating breaking cellular wall, vacuum decompression concentrates, and dialysis waits personated old man's production. art, causes multiple coenzyme inactivation and degraded to destroy, thereby has influenced the scope of its coenzyme kind and clinical practice.
Summary of the invention
The objective of the invention is for overcoming the weak point of prior art, a kind of preparation method and Coenzyme Complex medicine and clinical purposes of new Coenzyme Complex medicine are proposed, by changing the production technology of existing Coenzyme Complex, make that the coenzyme material in the former Coenzyme Complex drug products is increased to nine kinds from original four kinds; This Coenzyme Complex can protect various kinds of cell to avoid anoxia-induced apoptosis, keeps or recovers its homergy, makes it can be used for heart, operation on liver, and hepatic injury recovers, clinical treatments such as the cell injury recovery that ray or chemotherapy cause.
A kind of Coenzyme Complex medicament preparation that the present invention proposes may further comprise the steps:
1) fresh yeast being added 5-8 times of pH value is that PH3.0～5.0,0.01M～0.03M hac buffer are made the yeast cells suspension;
2) with the cell wall of high pressure homogenizer at 70～150Mpa pressure breaking yeast cells, temperature maintenance is at 0 ℃～20 ℃ when broken;
3) with the yeast cells suspension frozen centrifugation after the fragmentation, the frozen centrifugation condition is 0 ℃～5 ℃, and speed is 20000～35000CPM, 30～60 minutes time, obtains clear liquid;
4) use the activated carbon adsorption filter liquor, concentrated usefulness contains 40% alcoholic solution eluting of 2.0～3.5% ammonia;
5) adopt ion-exchange chromatography: desorbed solution is transferred PH7.5-9.0, and last anion-exchange column is with PH8.0～9.0,0.01～0.03M phosphate buffer eluant solution;
6) eluent is the 5000-10000 ultrafiltration through the molecular weight that dams;
7) ultrafiltrate removes H through nanofiltration or reverse osmosis
2O is concentrated into original volume 1/5-1/10, transfers PH2.5～3.5, adds 5～10 times of volumes ,-10 ℃～-20 ℃ cold acetones, collecting precipitation;
9) the degerming embedding of the water-soluble back of precipitation, lyophilizing.
Adopt the Coenzyme Complex medicine of above-mentioned preparation method, its coenzyme composition contains coenzyme A (CoA), nadide (NAD), glutathione (GSH), adenosine triphosphate (ATP) is characterized in that, also contains flavin mononucleotide (FMN) (FMN), flavin adenine dinucleotide (FAD) (FAD), gland two phosphorus (ADP), adenosine phosphate (AMP), ademetionine (SAM).
Above-mentioned Coenzyme Complex not only is used for acute, chronic hepatitis, idiopathic thrombocytopenic purpura, thrombocytopenia; as coronary atherosclerosis, chronic arterits, myocardial infarction, the weak oliguria that causes of renal function, uremic auxiliary therapeutic agent, also be used for the medicine of operation on heart, myocardial ischemia, cerebral anoxia, pesticide intoxication, atomic disease and the protection medicine of various histiocytic anoxia-induced apoptosis, physics and chemical damage.
Characteristics of the present invention and effect:
Preparation method of the present invention has used the high pressure homogenizer smudge cells to replace original heating breaking cellular wall; Use hyperfiltration technique to replace original dialysis; Use anti-dialysis and nanofiltration to replace original vacuum decompression to concentrate; Make the coenzyme material in the former Coenzyme Complex drug products be increased to nine kinds from original four kinds; and show that through zoopery new Coenzyme Complex can protect various kinds of cell to avoid anoxia, radical damage; keep or recover its homergy; make it can be used for heart, operation on liver; hepatic injury recovers, clinical treatments such as the cell injury recovery that ray, chemotherapy and operation cause.
The specific embodiment
Embodiment 1:
The soft tommy yeast adds 5 times of PH4.00.01M hac buffers and makes the yeast cells suspension; At 100Mpa pressure breaking yeast cells, keep cell suspension below 20 ℃ with high pressure homogenizer; .5 under ℃ condition with the yeast cells suspension frozen centrifugation after the fragmentation, speed is centrifugal 60 minutes of 20000CPM. clear liquid; Activated carbon adsorption, with the 40% cholamine solution stripping that contains 2% ammonia, stripping liquid is transferred PH8.0, and last Ambrite IRC50 ion exchange column is used PH8.5, and the 0.02M phosphoric acid liquid is towards the liquid eluting; Eluent concentrates through nanofiltration and removes H with the ultrafilter ultrafiltration of the molecular weight 8000 that dams, filter liquor
2O is concentrated into original volume 1/10, transfers PH3.5, adds 5 times of volumes ,-10 ℃ of cold acetone precipitations, and collecting precipitation is dissolved in the apirogen water, degerming embedding, lyophilizing.
Measure the concentration of each organic principle, degerming is freeze-up to be done.
The product analysis result:
| CoA | 120U/ml |
| NAD | 0.12mg/ml |
| ATP | 2.1mg/ml |
| ADP | 0.3mg/ml |
| AMP | 0.2mg/ml |
| FMN | 8ug/ml |
| FAD | 6ug/ml |
| GSH | 4.1mg/ml |
| SAM | 10.0ug/ml |
Embodiment 2:
The young beer yeast adds 6 times of PH3.0,0.03M acetate buffer solution, with high pressure homogenizer smudge cells under 70Mpa pressure, keeps cell breakage liquid below 10 ℃, and 30000CPM is centrifugal 45 minutes under 3 ℃ of conditions, gets clear liquid; It with the molecular weight that dams 8000 ultrafilter membrane ultrafiltration; Activated carbon adsorption is resolved with 40% cholamine that contains 3.0% ammonia; Desorbed solution is transferred AmbriteIRC 50 ion exchange column on the PH to 7.5, uses PH9.0,0.01M phosphate buffer eluting; Eluent is 5000 ultrafilter membrane ultrafiltration through the molecular weight that dams; Ultrafiltrate removes H with reverse osmosis
20 is concentrated into original volume 1/5, transfers PH2.5, adds the cold acetone of 10 times of volumes-15 ℃, and 0 ℃ is spent the night, next day collecting precipitation.Precipitation is dissolved in apirogen water, measures the concentration of each composition, degerming embedding, lyophilizing.
The product analysis result:
| CoA | 180U/ml |
| NAD | 0.22mg/ml |
| ATP | 2.3mg/ml |
| ADP | 0.4mg/ml |
| AMP | 0.5mg/ml |
| FMN | 2ug/ml |
| FAD | 4ug/ml |
| GSH | 3.3mg/ml |
| SAM | 18.0ug/ml |
Embodiment 3:
With the fresh food yeast, add 8 times of PH4.5, the 0.01M hac buffer; Use high pressure homogenizer, under 150Mpa pressure, broken yeast cell is kept broken liquid below 5 ℃; Centrifugal 35000cpm under 0 ℃ of condition obtained clear liquid in 30 minutes; Collect filter liquor; The activated carbon adsorption filter liquor is resolved with 40% ethanol that contains 3.5% ammonia; Desorbed solution is transferred PH to 9.0, last AmbriteIRC 50 ion exchange column, with PH8.0,0.03M phosphate buffer eluting, eluent is 9000 ultrafilter membrane ultrafiltration through the molecular weight that dams, ultrafiltrate through reverse osmosis concentration to original volume 1/8, transfer PH3.0, add the cold acetone of 8 times of volumes-20 ℃, next day collecting precipitation.Precipitation is dissolved in apirogen water, measures the concentration of each composition, degerming embedding, lyophilizing.
The product analysis result:
| The coenzyme title | Concentration |
| CoA | 120U/ml |
| NAD + | 0.32mg/ml |
| FMN | 8ug/ml |
| FAD | 6ug/ml |
| ATP | 1.0mg/ml |
| ADP | 0.3mg/ml |
| AMP | 0.5mg/ml |
| GSH | 2.1mg/ml |
| SAM | 5.0ug/ml |
Embodiment 4: fresh geotrichum candidum is added 5 times of PH3.0,0.01M acetate buffer solution; Under high pressure homogenizer 150Mpa pressure, smudge cells, broken liquid are kept below 15 ℃; Centrifugal 35 minutes of 4 ℃ of 30000cpm, clear liquid; Activated carbon adsorption, with 40% alcohol desorption that contains 2.0% ammonia, stripping liquid is transferred PH8.5, last Ambrite IRC 50 ion exchange column, use PH8.5,0.02M the phosphate buffer eluting, eluent is 5000 ultrafilter membrane ultrafiltration through the molecular weight that dams, and filtrate is condensed into 1/8 of original volume through nanofiltration, transfer PH3.5, add 10 times of-15 ℃ of cold acetones, put 0 ℃ and spend the night collecting precipitation.Be dissolved in a certain amount of apirogen water, measure each composition and through degerming embedding, lyophilizing.
The product analysis result:
| The coenzyme title | Concentration |
| CoA | 250U/ml |
| NAD | 0.41mg/ml |
| ATP | 0.5mg/ml |
| ADP | 0.23mg/ml |
| AMP | 0.22mg/ml |
| FAD | 7ug/ml |
| FMN | 10ug/ml |
| GSH | 1.5mg/ml |
| SAM | 6ug/ml |
Use Coenzyme Complex of the present invention, the protective effect to cardiac resuccitation (CPR) back rat thyroid is described as follows:
Use suffocate (succinylcholine) merge 0.5M ice chlorination potassium arrest liquid and cause the rat cardiac arrest, begin cardio-pulmonary resuscitation after 5 minutes.Experiment divides three groups: 1. matched group be sham operated rats (without suffocate, the rat of cardiac arrest and cardio-pulmonary resuscitation process); 2. conventional recovery group is 24 hours groups after the cardio-pulmonary resuscitation; 3. Coenzyme Complex experimental group.Conventional medicine recovery give simultaneously Coenzyme Complex (by one/10Kg) normal saline dilution pneumoretroperitoneum injects, recover and gave Isodose in back 12 hours again. every group of 8 rats.
(1) test rat blood serum thyroxine T
3, T
4The variation of thyrotropin is as shown in Table I:
Table 1 serum T 3, T4, TSH change (X ± S)
| Grouping | T3(ng/ml) | T4(Hg/dl) | TSH(μIU/ml) |
| The conventional recovery group of matched group shellfish section can organize | 0.56±0.06 0.23±0.05 * 0.29±0.4 *# | 5.11±0.39 1.55±0.35 * 2.45±0.58 *# | 1.61±0.40 1.44±0.35 1.52±0.24 |
With matched group ratio and matched group ratio
*P＜0.05, with routine recovery group than #P＜0.05.
The result shows: CPR can cause serum T
3, T
4, TSH concentration obviously descends, but conventional recovery group is starkly lower than the Coenzyme Complex group, two groups of T
3, T
4There was a significant difference.
(2) MDA (malonaldehyde) resultant in the parathyroid tissue, superoxide dismutase (SOD) vigor and Na
+-K
+The variation of-ATPase vigor is as shown in table 2;
MDA content in each group tissue of table 2, SOD vigor, Na+-K+-ATPase vigor (X ± S)
| Grouping | MDA content (nmol/mgprot) | SOD vigor (U/mgprot) | Na+-K+-ATPase vigor (μ mol/mgprot/hour) |
| The conventional recovery group of matched group shellfish section can organize | 0.49±0.09 1.52±0.21 * 1.78±0.20 *# | 11.14±0.98 7.82±1.14 * 9.64±1.04 *# | 1.07±0.17 0.45±0.17 0.58±0.10 *# |
With matched group ratio and matched group ratio
*P＜0.05, with routine recovery group than #P＜0.05.
The result shows: conventional recovery group parathyroid tissue MDA content is apparently higher than matched group, and apparently higher than the Coenzyme Complex group, and SOD and Na
+-K
+-Atpase vigor is lower than matched group, and significantly is lower than the Coenzyme Complex group.
(3) experimental mouse original position terminal transferase labelling (Tunel labelling) changes as shown in table 3:
Table 3 is respectively organized TUNEL. positive cell number average gray value (X ± S)
| Grouping | n | Positive cell number (individual) | Average gray value (%) |
| The conventional recovery group of matched group shellfish section can organize | 7 8 8 | 4.71±1.39 35.88±3.72 * 25.88±4.17 *# | 71.31±6.65 34.86±3.95 * 44.66±4.18 *# |
With contrast ratio
*P＜0.05, with routine recovery group than #P＜0.05.
The result shows: conventional recovery group Tunel positive cell is apparently higher than matched group, and is significantly higher than the Coenzyme Complex group, and its average gray value is starkly lower than matched group and is lower than the Coenzyme Complex group, and the difference between conventional group and Coenzyme Complex group has statistical significance.
(4) research of test rat Fas differential expression is as shown in table 4:
Table 4 is respectively organized FAS positive cell number average gray value (X ± S)
| Grouping | n | Positive cell number (individual). | Positive cell number average gray value (the average gray value (%) of X ± S) |
| The conventional recovery group of matched group shellfish section can organize | 7 8 8 | 7.00±1.31 37.00±4.03 * 27.25±4.21 *# | 65.32±5.72 26.41±3.82 * 36.69±4.62 *# |
With contrast ratio
*P＜0.05, with routine recovery group than #P＜0.05.
The result shows: Coenzyme Complex group Fas gene expression positive cell number and average gray and conventional recovery group relatively have significant difference.
More than four groups of tests show:
1. Coenzyme Complex has improved CPR rat SOD, Na
+-K
+-ATPase vigor improves the damage of histiocyte energy metabolism to antioxidant radical, and the resultant of lipid peroxidation catabolite MDA is lower than conventional recovery group in the exhibit tissue.
2. Coenzyme Complex has been reduced and has been transferred the expression of related gene Fas of dying, and shows as and transfer the obvious minimizing of cell (Tunel positive cell) of dying, thereby has alleviated the histiocyte damage that anoxia, perfusion cause, and rat behind the CPR is had protective effect.
3. Coenzyme Complex has alleviated the degree that recovery back rat blood serum thyroxin changes, and is of value to the prognosis that improves heart beating time-out and CPR.
Claims (4)
1, a kind of Coenzyme Complex medicament preparation may further comprise the steps:
1) fresh yeast being added 5-8 times of pH value is that PH3.0～5.0,0.01M～0.03M hac buffer are made the yeast cells suspension;
2) with the cell wall of high pressure homogenizer at 70～150Mpa pressure breaking yeast cells, temperature maintenance is at 0 ℃～20 ℃ when broken;
3) with the yeast cells suspension frozen centrifugation after the fragmentation, the frozen centrifugation condition is 0 ℃～5 ℃, and speed is 20000～35000CPM, 30～60 minutes time, obtains clear liquid;
4) use the activated carbon adsorption filter liquor, with the 40% alcoholic solution eluting that contains 2.0～3.5% ammonia;
5) adopt ion-exchange chromatography: desorbed solution is transferred PH7.5-9.0, and last anion-exchange column is with PH8.0～9.0,0.01～0.03M phosphate buffer eluant solution;
6) eluent is the 5000-10000 ultrafiltration through the molecular weight that dams;
7) ultrafiltrate removes H through nanofiltration or reverse osmosis
2O is concentrated into original volume 1/5-1/10, transfers PH2.5～3.5, adds 5～10 times of volumes ,-10 ℃～-20 ℃ cold acetones, collecting precipitation;
8) the degerming embedding of the water-soluble back of precipitation, lyophilizing.
2, preparation method as claimed in claim 1 is characterized in that, described fresh yeast bacterium is a kind of of fresh beer yeast, bakery yeast, edible yeast or geotrichum candidum.
3, the Coenzyme Complex medicine that obtains of preparation method according to claim 1, its coenzyme composition contains coenzyme A, nadide, glutathione, adenosine triphosphate, it is characterized in that, also contain flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), gland two phosphorus, adenosine phosphate, ademetionine; The concentration of described coenzyme composition is as follows:
FAD 2～20ug/ml
FMN 2～20ug/ml
ADP 0.1～0.5mg/ml
AMP 0.1～0.5mg/ml
SAM 1～20ug/ml
CoA 100～300U/ml、
NAD 0.1～0.5mg/ml、
ATP 0.1～2.5mg/ml
GSH 1～5mg/ml。
4, Coenzyme Complex medicine as claimed in claim 3 is characterized in that, the ratio of ATP, ADP, AMP is E in the described coenzyme:
5, a kind of Coenzyme Complex medicine as claimed in claim 3 is at preparation auxiliary treatment acute, chronic hepatitis, idiopathic thrombocytopenic purpura, thrombocytopenia; coronary atherosclerosis, chronic arterits, myocardial infarction, the weak oliguria that causes of renal function, uremic adjuvant therapy medicaments; preparation be used for operation on heart, myocardial ischemia, cerebral anoxia, pesticide intoxication, atomic disease medicine, and preparation be used for protecting myocardial cell, liver cell and neurocyte and be not subjected to anoxia-induced apoptosis, and the application in the protection medicine of physics and chemotherapy damage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410058086 CN1259925C (en) | 2004-08-13 | 2004-08-13 | Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410058086 CN1259925C (en) | 2004-08-13 | 2004-08-13 | Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1586502A CN1586502A (en) | 2005-03-02 |
| CN1259925C true CN1259925C (en) | 2006-06-21 |
Family
ID=34603316
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410058086 Expired - Lifetime CN1259925C (en) | 2004-08-13 | 2004-08-13 | Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1259925C (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101023968B (en) * | 2007-02-09 | 2010-05-19 | 上海大学 | Coenzyme composition preparing method |
| CN101984978B (en) * | 2010-09-30 | 2012-06-27 | 海南康永药品有限公司 | Method for preparing compound coenzyme medicine |
| CN102174512A (en) * | 2011-02-23 | 2011-09-07 | 武汉工业学院 | Process method for producing yeast polysaccharide and yeast nucleotide from yeast |
| CN103191145A (en) * | 2013-03-27 | 2013-07-10 | 南京理工大学 | Application of adenosine and derivatives thereof in prevention and treatment of medicament-induced liver injury |
| CN103341155B (en) * | 2013-07-17 | 2015-03-11 | 北京双鹭药业股份有限公司 | Stable compound coenzyme preparation and preparation method as well as application thereof |
| CN104623626A (en) * | 2013-07-17 | 2015-05-20 | 北京双鹭药业股份有限公司 | Stable compound coenzyme preparation as well as preparation method and applications thereof |
| CN104844487A (en) * | 2015-04-05 | 2015-08-19 | 杭州蓝然环境技术有限公司 | Method for decoloring methionine crystallization mother liquor by adopting membrane integration technology |
| CN104892710B (en) | 2015-05-19 | 2017-09-29 | 邦泰生物工程(深圳)有限公司 | A kind of method for purifying reduced form β NADHs |
| CN104876994B (en) * | 2015-05-19 | 2017-11-03 | 邦泰生物工程(深圳)有限公司 | A kind of method for purifying oxidized form β NADHs |
| CN111202745B (en) * | 2020-03-06 | 2021-09-10 | 广东药科大学 | Application of FAD in preparation of medicine for inhibiting or treating cardiovascular system diseases |
| WO2022082487A1 (en) * | 2020-10-21 | 2022-04-28 | 朱上翔 | Anti-aging composite nutrient containing coenzymes and bioactive hydrogen, preparation method therefor and application thereof |
-
2004
- 2004-08-13 CN CN 200410058086 patent/CN1259925C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CN1586502A (en) | 2005-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1259925C (en) | Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use | |
| Huang et al. | Chiroinositol deficiency and insulin resistance. III. Acute glycogenic and hypoglycemic effects of two inositol phosphoglycan insulin mediators in normal and streptozotocin-diabetic rats in vivo | |
| Hwang et al. | Hypoglycemic effect of crude exopolysaccharides produced by a medicinal mushroom Phellinus baumii in streptozotocin-induced diabetic rats | |
| Sprott et al. | Novel polar lipids of halophilic eubacterium Planococcus H8 and archaeon Haloferax volcanii | |
| Green | The Mode of Action of Sulphanilamide: With Special Reference to a Bacterial Growth-Stimulating Factor (“P” Factor) Obtained from Br. Abortus and other Bacteria | |
| US20060270626A1 (en) | Crude exopolysaccharides produced from phellinus baumii mycelium having hypoglycemic activity and preparation method thereof | |
| Quinlan et al. | Group translocation of the ribose moiety of inosine by vesicles of plasma membrane from T (3 cells transformed by Simian virus 40. | |
| Schlenk et al. | Enzymatic activity of yeast cell ghosts produced by protein action on the membranes | |
| Winkler et al. | Acquisition of glucose by Rickettsia prowazekii through the nucleotide intermediate uridine 5'-diphosphoglucose | |
| CN104623626A (en) | Stable compound coenzyme preparation as well as preparation method and applications thereof | |
| US4446064A (en) | Insulin mediator substance | |
| US4190649A (en) | Polyribonucleotides capable of promoting the genesis of leucocytes and blood platelets | |
| Clegg | Interrelationships between water and cell metabolism in Artemia cysts. IX. Evidence for organization of soluble cytoplasmic enzymes | |
| Lee et al. | Effect of culture conditions on the structure of Streptococcus pneumoniae type 19A (57) capsular polysaccharide | |
| Opekarová et al. | Isolation and Properties of an Arginine‐Binding Protein from Saccharomyces cerevisiae | |
| CN111117917B (en) | Primary tetraodotoxin-producing globefish co-habitat and application thereof | |
| Petricciani et al. | Galactose utilization in galactosemia | |
| Mortlock et al. | [8] 2-ketopentoses | |
| Reizer et al. | Transport of α-aminoisobutyric acid by Streptococcus pyogenes and its derived L-form | |
| CN1090602A (en) | Pseudomonos aeruginosa MSHA fimbria strain and foundation thereof | |
| HORI et al. | Inhibition in Vitro of Growth of Rat Ascites Tumor Cells by Bovine Liver Extract Purification and Property of the Actice Principle | |
| Gimsing et al. | [1] Determination of cobalamins in biological material | |
| Osumi et al. | Physiological activities of oligosaccharides derived from marine algae Porphyra yezoensis Porphyran | |
| JP2714663B2 (en) | Intestinal beneficial bacteria growth promoter | |
| Connell et al. | Incorporation of C14 into bacterial peptides |
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 | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20050302 Assignee: BEIJING SL PHARMACEUTICAL Co.,Ltd. Assignor: Aolute Biomedicines Research & Development Co.,Ltd. Beijing|Beijing Suliman Medicine Technology Co.,Ltd.|HAINAN KANGYONG PHARMACEUTICALS Co.,Ltd. Contract record no.: 2013990000759 Denomination of invention: Process for preparing compound coemzyme medicine and its compound coemzyme medicine and clinical use Granted publication date: 20060621 License type: Exclusive License Record date: 20131113 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CX01 | Expiry of patent term |
Granted publication date: 20060621 |
|
| CX01 | Expiry of patent term |