CN107325153A - A kind of antihypertensive active peptide Citn Hyp Pro and application and pharmaceutical composition - Google Patents
A kind of antihypertensive active peptide Citn Hyp Pro and application and pharmaceutical composition Download PDFInfo
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- CN107325153A CN107325153A CN201710620147.4A CN201710620147A CN107325153A CN 107325153 A CN107325153 A CN 107325153A CN 201710620147 A CN201710620147 A CN 201710620147A CN 107325153 A CN107325153 A CN 107325153A
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- citn
- active peptide
- pro
- hyp
- antihypertensive
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 38
- 230000003276 anti-hypertensive effect Effects 0.000 title claims abstract description 20
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 8
- 241000219112 Cucumis Species 0.000 claims abstract description 12
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 claims abstract description 12
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims abstract description 8
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 239000003814 drug Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 4
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 235000013305 food Nutrition 0.000 claims description 2
- 235000013373 food additive Nutrition 0.000 claims description 2
- 239000002778 food additive Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 241000700159 Rattus Species 0.000 abstract description 17
- 230000036772 blood pressure Effects 0.000 abstract description 16
- 230000000694 effects Effects 0.000 abstract description 15
- 206010020772 Hypertension Diseases 0.000 abstract description 10
- 238000003304 gavage Methods 0.000 abstract description 9
- 125000000266 alpha-aminoacyl group Chemical group 0.000 abstract description 5
- 230000001631 hypertensive effect Effects 0.000 abstract description 5
- 206010010356 Congenital anomaly Diseases 0.000 abstract description 4
- GIOMWAZJQDYWEU-UHFFFAOYSA-N Hydroxyprolyl-Proline Chemical compound C1C(O)CNC1C(=O)N1C(C(O)=O)CCC1 GIOMWAZJQDYWEU-UHFFFAOYSA-N 0.000 abstract description 4
- 230000037396 body weight Effects 0.000 abstract description 4
- 230000001629 suppression Effects 0.000 abstract description 4
- 230000000857 drug effect Effects 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 description 13
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- JIGUQPWFLRLWPJ-UHFFFAOYSA-N Ethyl acrylate Chemical compound CCOC(=O)C=C JIGUQPWFLRLWPJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 6
- PNJWIWWMYCMZRO-UHFFFAOYSA-N pent‐4‐en‐2‐one Natural products CC(=O)CC=C PNJWIWWMYCMZRO-UHFFFAOYSA-N 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 6
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 5
- 229940024606 amino acid Drugs 0.000 description 5
- 235000001014 amino acid Nutrition 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- 230000002194 synthesizing effect Effects 0.000 description 5
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 4
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 108010025306 histidylleucine Proteins 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- MMFKFJORZBJVNF-UWVGGRQHSA-N His-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](N)CC1=CN=CN1 MMFKFJORZBJVNF-UWVGGRQHSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 239000007832 Na2SO4 Substances 0.000 description 3
- DOFAQXCYFQKSHT-SRVKXCTJSA-N Val-Pro-Pro Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DOFAQXCYFQKSHT-SRVKXCTJSA-N 0.000 description 3
- 229960000830 captopril Drugs 0.000 description 3
- FAKRSMQSSFJEIM-RQJHMYQMSA-N captopril Chemical compound SC[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O FAKRSMQSSFJEIM-RQJHMYQMSA-N 0.000 description 3
- 238000005352 clarification Methods 0.000 description 3
- 239000004519 grease Substances 0.000 description 3
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 3
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 description 3
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 238000004617 QSAR study Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- -1 hydroxyl tert-butyl group Chemical group 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 229920000139 polyethylene terephthalate Polymers 0.000 description 2
- 239000005020 polyethylene terephthalate Substances 0.000 description 2
- 239000011265 semifinished product Substances 0.000 description 2
- 229940126586 small molecule drug Drugs 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000035488 systolic blood pressure Effects 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- AAXWBCKQYLBQKY-IRXDYDNUSA-N (2s)-2-[[(2s)-2-[(2-benzamidoacetyl)amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-4-methylpentanoic acid Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)C=1C=CC=CC=1)C1=CN=CN1 AAXWBCKQYLBQKY-IRXDYDNUSA-N 0.000 description 1
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- FZRCKLPSHGTOAU-UHFFFAOYSA-N 6-amino-1,4-dimethylcyclohexa-2,4-diene-1-carbaldehyde Chemical compound CC1=CC(N)C(C)(C=O)C=C1 FZRCKLPSHGTOAU-UHFFFAOYSA-N 0.000 description 1
- 208000028185 Angioedema Diseases 0.000 description 1
- XPCFTKFZXHTYIP-PMACEKPBSA-N Benazepril Chemical compound C([C@@H](C(=O)OCC)N[C@@H]1C(N(CC(O)=O)C2=CC=CC=C2CC1)=O)CC1=CC=CC=C1 XPCFTKFZXHTYIP-PMACEKPBSA-N 0.000 description 1
- 0 C*C(CCCI)[I+](C1=CCC(*)C[C@@]1C([*@@](CCCC1)[C@]1C(*)=C)=C*O)=O Chemical compound C*C(CCCI)[I+](C1=CCC(*)C[C@@]1C([*@@](CCCC1)[C@]1C(*)=C)=C*O)=O 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 108010061435 Enalapril Proteins 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 description 1
- 108010007859 Lisinopril Proteins 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 108091000041 Phosphoenolpyruvate Carboxylase Proteins 0.000 description 1
- RWCOTTLHDJWHRS-YUMQZZPRSA-N Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 RWCOTTLHDJWHRS-YUMQZZPRSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960004530 benazepril Drugs 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229910002114 biscuit porcelain Inorganic materials 0.000 description 1
- 230000004531 blood pressure lowering effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229960002173 citrulline Drugs 0.000 description 1
- 235000013477 citrulline Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 235000015177 dried meat Nutrition 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 229960000873 enalapril Drugs 0.000 description 1
- GBXSMTUPTTWBMN-XIRDDKMYSA-N enalapril Chemical compound C([C@@H](C(=O)OCC)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(O)=O)CC1=CC=CC=C1 GBXSMTUPTTWBMN-XIRDDKMYSA-N 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 108010016268 hippuryl-histidyl-leucine Proteins 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 229960002394 lisinopril Drugs 0.000 description 1
- RLAWWYSOJDYHDC-BZSNNMDCSA-N lisinopril Chemical compound C([C@H](N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(O)=O)C(O)=O)CC1=CC=CC=C1 RLAWWYSOJDYHDC-BZSNNMDCSA-N 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- HQEIPVHJHZTMDP-JEDNCBNOSA-N methyl (2s)-pyrrolidine-2-carboxylate;hydrochloride Chemical compound Cl.COC(=O)[C@@H]1CCCN1 HQEIPVHJHZTMDP-JEDNCBNOSA-N 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- NONJJLVGHLVQQM-JHXYUMNGSA-N phenethicillin Chemical compound N([C@@H]1C(N2[C@H](C(C)(C)S[C@@H]21)C(O)=O)=O)C(=O)C(C)OC1=CC=CC=C1 NONJJLVGHLVQQM-JHXYUMNGSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- 238000012216 screening Methods 0.000 description 1
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- 238000007873 sieving Methods 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000002618 waking effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0819—Tripeptides with the first amino acid being acidic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The present invention relates to a kind of antihypertensive active peptide Citn Hyp Pro and application and pharmaceutical composition.The amino acid sequence of the active peptide is：Melon aminoacyl hydroxyprolyl- proline, with structure shown in Formulas I.The IC of Angiotensin-Converting (ACE) activity suppression of active peptide of the present invention50For 69.54 μm of ol/L, minimum point 193mmHg is dropped to from 224mmHg using blood pressure after congenital Hypertensive Rats of the dosage gavage of 1.0mg/kg body weight 3.5 hours, the constant drug effect time is 2 hours.
Description
Technical field
The invention belongs to biomedicine field, more particularly, to a kind of antihypertensive active peptide Citn-Hyp-Pro and
Using, and a kind of antihypertensive pharmaceutical composition.
Background technology
《Chinese cardiovascular disease report 2015》Point out that the ill rate of cardiovascular disease of China is in and continue ascent stage.Hypertension
Be angiocardiopathy Major Risk Factors into common recognition, it be the promotion that persistently rises of China's angiocardiopathy illness rate because
Element.Ministry of Public Health's statistics in 2014 shows that the hypertension sufferer rate of China is 25.5%, and some provinces reach 27.9%, the whole nation
Hyperpietic's number is estimated up to 2.7 hundred million.
At present, many drugs for hypertension are considered as the medicine of a clinical line, wherein Angiotensin-Converting
Inhibitor (ACEI) is the best medicine of clinical effectiveness, and how general such as enalapril, captopril, Benazepril, lisinopril, group be
Profit etc..But they still have certain side effect, such as dry cough, fash, angioedema and injury of kidney.
ACEI class medicines are a class peptides and modified amino acid class material, and natural albumen is abundant peptide resource.Cause
This, scientists are endeavoured to prepare ACEI active peptides by the way of enzyme hydrolysis and fermentation always, to make hyperpietic
Benefit.As two have anti-hypertension bioactivity tripeptides " VPP " (Val-Pro-Pro, VPP) and " IPP " (Ile-Pro-Pro,
IPP) it is accredited out.But, high, the repeated low, bioavailability of this preparation method cost is low, in the urgent need to development is a kind of
New replacement production technology.These researchs are all based on the peptide information contained in protein, although with the increase of peptide molecular weight,
Its structure will vary, and activity will also have a potentiality further, but be due to that the molecular sieving effect of intestinal mucosa is rich in it
Peptase, constrains and utilizes the active development of resources ACEI peptides small-molecule substances of native protein.Therefore, chemical modification peptide how is evaded
The side effect of class or amino acids ACEI small-molecule drugs and avoid internal peptase hydrolysis and absorbed intact, as exploitation safety
The ACEI small-molecule drug study hotspots having no side effect.
The content of the invention
It is an object of the invention to provide a kind of antihypertensive active peptide Citn-Hyp-Pro and application and pharmaceutical composition.
The first aspect of the present invention is to provide a kind of antihypertensive active peptide Citn-Hyp-Pro, the amino of the active peptide
Acid sequence is：Melon aminoacyl-hydroxyprolyl--proline, with structure shown in Formulas I.Referred to as Citn-Hyp-Pro (Citrulline
(Citn)-Hydroxyproline(Hyp)-Proline(Pro))。
The present invention is from quantitative structure activity relationship (the QSAR)s, using food-borne rare amino acid, with reference to antihypertensive active
The structure effect feature of PEPC end Pro tolerance peptide enzyme hydrolysis, designs a series of small-molecule substances, passes through CLC Drug Discovery
The chemical characteristic of Workbench analyses design small-molecule substance and ACE interaction, is primarily determined that with antihypertensive peptide
Molecule, is prepared by liquid phase synthesizing method, is further verified by vitro and in vivo bioactivity, and screening determines to have relatively strong anti-height
The synthetic peptide of blood pressure activity.The antihypertensive active peptide can be made by the conventional method of this area, such as liquid phase synthesizing method.
The second aspect of the present invention provide described antihypertensive active peptide Citn-Hyp-Pro prepare it is antihypertensive
Application in medicine and food.
The third aspect of the present invention provides a kind of antihypertensive pharmaceutical composition, including active ingredient and auxiliary material, described
Active ingredient include described antihypertensive active peptide Citn-Hyp-Pro.
According to the present invention, the auxiliary material can be conventional various pharmaceutic adjuvants or food additives.
The present invention has following advantage compared with prior art：
The present invention proposes a kind of different active peptide.Prior art passes through external proteolysis or micro- from native protein
Biofermentation separation obtains active peptide, can further be hydrolyzed by the peptase being rich in intestinal mucosa, reduces it actual in vivo
Effect.The active peptide of the present invention is made up of rare amino acid and conventional amino acid, in native protein and in the absence of such sequence
Row are constituted.Therefore, active peptide of the invention can escape the further hydrolysis of vivo protein enzyme, it is ensured that its integrality absorbed
The stability acted in vivo.
The IC of Angiotensin-Converting (ACE) activity suppression of active peptide of the present invention50For 69.54 μm of ol/L, use
The congenital Hypertensive Rats of dosage gavage of 1.0mg/kg body weight after 3.5 hours blood pressure drop to minimum point from 224mmHg
193mmHg, the constant drug effect time is 2 hours.
Other features and advantages of the present invention will be described in detail in subsequent embodiment part.
Brief description of the drawings
By the way that exemplary embodiment of the invention is described in more detail with reference to accompanying drawing, it is of the invention above-mentioned and its
Its purpose, feature and advantage will be apparent.
Fig. 1 is Citn-Hyp-Pro to congenital hypertensive rat blood pressure function analysis laboratory test results figure.
Embodiment
With reference to specific embodiment, the present invention will be further described in detail, but implements the invention is not restricted to following
Example.In following examples, when being not particularly illustrated, " % " refers both to mass percent.
Embodiment 1
The synthesis of Citn-Hyp-Pro tripeptides.Antihypertensive active peptide of the present invention is synthesized by artificial chemistry, concrete operations
It is as follows：
Polypeptide of the present invention is synthesized using Liquid phase peptides synthesis method, by a certain amount of N- tertbutyloxycarbonyls citrulling methyl esters
(Boc-Cit) inserted with hydroxysuccinimide (Hosu) in round-bottomed flask, add tetrahydrofuran (THF) and be dissolved to clarification, after
It is continuous to add dicyclohexylcarbodiimide (DCC), it is gently mixed, reaction is stayed overnight.
Further, above-mentioned reaction solution is filtered by vacuum, ethyl acrylate (EA) and H is added in filtrate2O is carried out
Chromatography.Supernatant liquid is drawn, using NaHCO3/H2O is washed 2 times, then with citric acid/H2O is washed 2 times, the washing of saturation NaCl solution
2 times, Na2SO4Dry, the grease that Rotary Evaporators are evaporated acquisition is N- tertbutyloxycarbonyl melon aminoacyl succimides (Boc-
Citn-osu)。
Further, tert-butyl group hydroxyl dried meat is added in N- tertbutyloxycarbonyl melon aminoacyl succimides (Boc-Cit-osu)
Propylhomoserin (Hyp (tbu)), is dissolved in tetrahydrofuran aqueous solution (THF:H2O=2：1) to clarifying, reaction is stirred at room temperature in alkalescence condition
2 hours.Add the absolute ether of 6-8 times of volume while stirring in reaction solution, chromatographed, 4000rpm is centrifuged 3 minutes.Abandon
Ether repeated washing is used after supernatant 5 times, precipitation obtains white solid matter and be dried in vacuo obtaining N- tertbutyloxycarbonyl melon ammonia
The acyl hydroxyprolyl- tert-butyl ester (Boc-Citn Hyp (tbu)).
Further, by a certain amount of N- tertbutyloxycarbonyls melon aminoacyl hydroxyprolyl- tert-butyl ester (Boc-Citn Hyp
(tbu)) inserted with proline methyl ester hydrochloride (Pro-Ome.HCl) in round-bottomed flask, add dimethylformamide (DMF) dissolving
To clarifying, dicyclohexylcarbodiimide (DCC) is continuously added, is gently mixed, reaction is stayed overnight.
Further, confirm whether reaction is complete by thin-layer chromatography (TCL).Above-mentioned reaction solution is filtered by vacuum,
Ethyl acrylate (EA) and H are added in filtrate2O is chromatographed.Supernatant liquid is drawn, using NaHCO3/H2O is washed 2 times, then
With citric acid/H2O is washed 2 times, and saturation NaCl solution is washed 2 times, Na2SO4Dry, Rotary Evaporators are evaporated the grease of acquisition
For N- tertbutyloxycarbonyl melon aminoacyl tert-butyl group hydroxyprolyl- proline methyl esters (Boc-Citn-Hyp (tbu)-Pro-Ome).
Further, in N- tertbutyloxycarbonyl melon aminoacyl hydroxyl tert-butyl group prolyl proline methyl esters (Boc-Citn-Hyp
(tbu)-Pro-Ome) in add methanol (MeOH) and tetrahydrofuran (THF) (1:1) solution is stirred reaction, after solution clarification
Add lithium hydroxide (LiOH)/H2O regulation pH ≈ 13, are kept for 2 hours, confirm whether reaction is complete by thin-layer chromatography (TCL).
Reaction solution is inserted into separatory funnel, polyethylene terephthalate (PET) cyclic washing is added 2 times, interception lower floor liquid.
Ethyl acrylate is added in lower floor's liquid, is adjusted with 2N HCl after solution to acidity, separatory funnel layering is placed in, passes through thin layer
Chromatograph (TCL) to confirm, intercept supernatant liquid.By supernatant liquid citric acid/H2O is washed 2 times, and saturation NaCl solution is washed 2 times,
Na2SO4Dry, the grease that Rotary Evaporators are evaporated acquisition is N- tertbutyloxycarbonyl melon aminoacyl tert-butyl group hydroxyprolyl- proline
(Boc-Citn-Hyp(tbu)-Pro-OH)。
In N- tertbutyloxycarbonyl melon aminoacyl tert-butyl group hydroxyprolyl- proline (Boc-Citn-Hyp (tbu)-Pro-OH)
4N hydrogen chloride gas/ethyl acrylate is added, clarification is dissolved to, reaction 2 hours is stirred at room temperature.Add 6- while stirring in reaction solution
The absolute ether of 8 times of volumes, is chromatographed, 4000rpm is centrifuged 3 minutes.Abandon after supernatant and use ether repeated washing 5 times, separate out
To white solid matter be dried in vacuo obtaining melon aminoacyl hydroxyprolyl- proline tripeptides (Citn-Hyp-Pro) semifinished product.
Further, semifinished product passes through semi-preparative reverse-phase high performance liquid chromatography (reversed-phase column：30 × 250 millimeters of Yi Lite
C18Post；Mobile phase (acetonitrile of A liquid 100% (ACN), B liquid 100%H2O), linear gradient 14%~80%；The ml/min of flow velocity 3)
Eluting peak is separated and collected, it is standby after freezing.
Embodiment 2
Extracorporeal blood vessel Converting Enzyme (ACE) active suppression test.
Horse urea acyl histidyl- leucine (hippuryl-L-histidyl-L-leucine, HHL) is under the catalysis of ACE enzymes
Fast decoupled produces hippuric acid (Hippuric Acid, HA) and dipeptides histidyl-leucine (HL), adds ACE enzyme levels
After agent, the activity of ACE enzymes is suppressed, and HA and HL growing amount are reduced, and ACE is extracted by rabbit lung in the present embodiment, enzyme activity
For 3.19mU/mL, developed the color by DAB, HA growing amounts are determined using spectrophotometer method, analyze ACE enzyme activity.Ethyl acetate is extracted
Hippuric acid in reactant, then reacts in acetic anhydride with the pyridine solution (DAB developers) containing paradime thylaminobenzaldehyde
Bisque compound is generated, directly in its OD value of 459nm colorimetric estimations, ACE enzyme inhibitors are evaluated to ACE enzymes by following equation
Inhibiting rate.Concentration (the IC of synthesis tripeptides needed for being suppressed with 50% ACE enzymatic activitys50) define the ACE suppressions for synthesizing tripeptides
System activity.
ACE inhibitory activity (%)=[(ODcontrol-ODsample)/(ODcontrol-ODblank)] × 100%
Specific reaction system and condition are shown in Table 1
Table 1
The present embodiment method measures the active kyrine of the present invention to ACE inhibitory activity IC50For 69.54 μm of ol/L.
Embodiment 3
The internal drop test of congenital Hypertensive Rats (SHR).
Using the intelligent non-invasive blood pressure instrument of SoftronBP-98A types rat, the systolic pressure for covering tail method measure rat is utilized
(SBP)。
Under SHR rats (spontaneous hypertensive rat) waking state, mouse is placed in mouse bag first, constant temperature is kept,
The method that set tail determines tail vein blood pressure is carried out using the intelligent non-invasive blood pressure instrument of SoftronBP-98A types rat, rat is determined
Systolic pressure (SBP).Start to determine the blood pressure of rat every other day in experiment the last week, experimental record is started after rat stabilization is adapted to.
The rat blood pressure before gavage is first determined, then 1.0mg/kg body weight doses carry out sample (active kyrine of embodiment 1, Citn-
Hyp-Pro) gavage, the isometric physiological saline of blank control group gavage (Saline), drug control group gavage 10mg/kg body weight
Drug for hypertension captopril (captopril), is carried out continuously monitoring of blood pressure 5 hours, after sample gavage every 30 minutes it is continuous
Record rat blood pressure.Each test point determines the blood pressure of 3 rats, about 1 minute interval time determined every time, takes 3 surveys
The average value of definite value as the test point rat blood pressure, as a result as shown in Figure 1.
Fig. 1 is the blood pressure situation after synthesizing activity peptide group rat oral gavage is administered, and its data measured is soft with SPSS systems
Part carries out system processing, using the t methods of inspection.From experimental result, compared with drug control group, the pressure reduction effect of synthetic peptide
It is more sluggish 90 minutes or so than medicine group, while Effect time is relatively short, about 2 hours.Synthesizing activity peptide gavage 3.5 hours
Afterwards, SHR blood pressures drop significantly to minimum point, after gavage 5 hours, blood pressure rises to initial pressure value.Illustrate active peptide of the present invention
(Citn-Hyp-Pro) there is preferable blood pressure lowering effect.
It is described above various embodiments of the present invention, described above is exemplary, and non-exclusive, and
It is not limited to disclosed each embodiment.In the case of without departing from the scope and spirit of illustrated each embodiment, for this skill
Many modifications and changes will be apparent from for the those of ordinary skill in art field.
Claims (4)
1. a kind of antihypertensive active peptide Citn-Hyp-Pro, it is characterised in that the amino acid sequence of the active peptide is：Melon ammonia
Acyl-hydroxyprolyl--proline, with structure shown in Formulas I：
2. the antihypertensive active peptide Citn-Hyp-Pro described in claim 1 is in antihypertensive medicine and food is prepared
Using.
3. a kind of antihypertensive pharmaceutical composition, including active ingredient and auxiliary material, it is characterised in that described active ingredient bag
Include antihypertensive active peptide Citn-Hyp-Pro as claimed in claim 1.
4. pharmaceutical composition according to claim 3, wherein, the auxiliary material is pharmaceutic adjuvant or food additives.
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| CN108484720A (en) * | 2018-05-14 | 2018-09-04 | 南京大学 | A kind of antihypertensive active peptide Orn-Pro-Hyp and application and pharmaceutical composition |
| CN112694429A (en) * | 2020-12-29 | 2021-04-23 | 江苏医药职业学院 | Polypeptide and application thereof in preparing ACE inhibitor or blood pressure lowering product |
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