CN105777866A - 一种抗高血压活性肽及制备 - Google Patents
一种抗高血压活性肽及制备 Download PDFInfo
- Publication number
- CN105777866A CN105777866A CN201610225861.9A CN201610225861A CN105777866A CN 105777866 A CN105777866 A CN 105777866A CN 201610225861 A CN201610225861 A CN 201610225861A CN 105777866 A CN105777866 A CN 105777866A
- Authority
- CN
- China
- Prior art keywords
- piperidines
- minutes
- dimethylformamide
- proline
- pro
- 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.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 32
- 206010020772 Hypertension Diseases 0.000 title claims abstract description 19
- 230000000975 bioactive effect Effects 0.000 title claims abstract description 10
- 238000000034 method Methods 0.000 title claims description 6
- 239000003814 drug Substances 0.000 claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 150000001413 amino acids Chemical group 0.000 claims abstract description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical group CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 51
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 20
- 150000003053 piperidines Chemical class 0.000 claims description 19
- 230000008878 coupling Effects 0.000 claims description 18
- 238000010168 coupling process Methods 0.000 claims description 18
- 238000005859 coupling reaction Methods 0.000 claims description 18
- 230000003276 anti-hypertensive effect Effects 0.000 claims description 16
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 10
- 239000012317 TBTU Substances 0.000 claims description 10
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 10
- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 claims description 10
- 238000010511 deprotection reaction Methods 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- -1 fluorenylmethyloxycarbonyl proline Chemical compound 0.000 claims description 9
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 4
- 229960000310 isoleucine Drugs 0.000 claims description 4
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 4
- 235000013305 food Nutrition 0.000 claims description 3
- 238000004007 reversed phase HPLC Methods 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- 239000004474 valine Substances 0.000 claims description 3
- 239000012059 conventional drug carrier Substances 0.000 claims description 2
- 125000005519 fluorenylmethyloxycarbonyl group Chemical group 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000011347 resin Substances 0.000 claims description 2
- 229920005989 resin Polymers 0.000 claims description 2
- ATHHXGZTWNVVOU-UHFFFAOYSA-N N-methylformamide Chemical compound CNC=O ATHHXGZTWNVVOU-UHFFFAOYSA-N 0.000 claims 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 claims 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 claims 1
- 239000004615 ingredient Substances 0.000 claims 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 claims 1
- 235000013372 meat Nutrition 0.000 claims 1
- 229940126701 oral medication Drugs 0.000 claims 1
- 230000002265 prevention Effects 0.000 claims 1
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 abstract description 16
- 102000004190 Enzymes Human genes 0.000 abstract description 16
- 108090000790 Enzymes Proteins 0.000 abstract description 16
- 230000036772 blood pressure Effects 0.000 abstract description 15
- 230000000694 effects Effects 0.000 abstract description 13
- 238000001727 in vivo Methods 0.000 abstract description 6
- 230000015572 biosynthetic process Effects 0.000 abstract description 5
- 230000004071 biological effect Effects 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 3
- 238000007796 conventional method Methods 0.000 abstract description 2
- 238000012795 verification Methods 0.000 abstract description 2
- YZPRRPLRFWMTMZ-NAEHWQOXSA-N (2S)-2-amino-3-methylbutanoic acid (2S,3S)-2-amino-3-methylpentanoic acid (2S)-pyrrolidine-2-carboxylic acid Chemical group CC(C)[C@H](N)C(O)=O.OC(=O)[C@@H]1CCCN1.OC(=O)[C@@H]1CCCN1.OC(=O)[C@@H]1CCCN1.OC(=O)[C@@H]1CCCN1.CC[C@H](C)[C@H](N)C(O)=O YZPRRPLRFWMTMZ-NAEHWQOXSA-N 0.000 abstract 1
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 abstract 1
- 241000700159 Rattus Species 0.000 description 13
- 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 7
- 230000007423 decrease Effects 0.000 description 6
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 108010025306 histidylleucine Proteins 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 3
- 239000005541 ACE inhibitor Substances 0.000 description 3
- 102400000345 Angiotensin-2 Human genes 0.000 description 3
- 101800000733 Angiotensin-2 Proteins 0.000 description 3
- 101710129690 Angiotensin-converting enzyme inhibitor Proteins 0.000 description 3
- 101710086378 Bradykinin-potentiating and C-type natriuretic peptides Proteins 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
- CZGUSIXMZVURDU-JZXHSEFVSA-N Ile(5)-angiotensin II Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=[NH2+])NC(=O)[C@@H]([NH3+])CC([O-])=O)C(C)C)C1=CC=C(O)C=C1 CZGUSIXMZVURDU-JZXHSEFVSA-N 0.000 description 3
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 3
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 3
- 229950006323 angiotensin ii Drugs 0.000 description 3
- 229940044094 angiotensin-converting-enzyme inhibitor Drugs 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 230000035488 systolic blood pressure Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 101800004538 Bradykinin Proteins 0.000 description 2
- 102400000967 Bradykinin Human genes 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 102000007079 Peptide Fragments Human genes 0.000 description 2
- 108010033276 Peptide Fragments Proteins 0.000 description 2
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 210000002565 arteriole Anatomy 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 229910021538 borax Inorganic materials 0.000 description 2
- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 210000005086 glomerual capillary Anatomy 0.000 description 2
- 230000001434 glomerular Effects 0.000 description 2
- 230000001631 hypertensive effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 230000008327 renal blood flow Effects 0.000 description 2
- 235000010339 sodium tetraborate Nutrition 0.000 description 2
- BSVBQGMMJUBVOD-UHFFFAOYSA-N trisodium borate Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-] BSVBQGMMJUBVOD-UHFFFAOYSA-N 0.000 description 2
- 230000002792 vascular Effects 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
- 208000028185 Angioedema Diseases 0.000 description 1
- 102000008873 Angiotensin II receptor Human genes 0.000 description 1
- 108050000824 Angiotensin II receptor Proteins 0.000 description 1
- 102400000344 Angiotensin-1 Human genes 0.000 description 1
- 101800000734 Angiotensin-1 Proteins 0.000 description 1
- 229940127291 Calcium channel antagonist Drugs 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 102000018389 Exopeptidases Human genes 0.000 description 1
- 108010091443 Exopeptidases Proteins 0.000 description 1
- 208000022461 Glomerular disease Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 208000019025 Hypokalemia Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 206010047139 Vasoconstriction Diseases 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- ORWYRWWVDCYOMK-HBZPZAIKSA-N angiotensin I Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 ORWYRWWVDCYOMK-HBZPZAIKSA-N 0.000 description 1
- 239000002333 angiotensin II receptor antagonist Substances 0.000 description 1
- 229940126317 angiotensin II receptor antagonist Drugs 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000000480 calcium channel blocker Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000023852 carbohydrate metabolic process Effects 0.000 description 1
- 235000021256 carbohydrate metabolism Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000011128 cardiac conduction Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 239000002934 diuretic Substances 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 210000003725 endotheliocyte Anatomy 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 229940125532 enzyme inhibitor Drugs 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 231100000852 glomerular disease Toxicity 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 230000000004 hemodynamic effect Effects 0.000 description 1
- 108010016268 hippuryl-histidyl-leucine Proteins 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000011587 new zealand white rabbit Methods 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 208000024896 potassium deficiency disease Diseases 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000029865 regulation of blood pressure Effects 0.000 description 1
- 230000036454 renin-angiotensin system Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000011699 spontaneously hypertensive rat Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 108010015385 valyl-prolyl-proline Proteins 0.000 description 1
- 230000025033 vasoconstriction Effects 0.000 description 1
- 230000024883 vasodilation Effects 0.000 description 1
- 235000020795 whole food diet Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
一种抗高血压活性肽,其氨基酸序列为:异亮氨酸‑脯氨酸‑脯氨酸‑缬氨酸‑脯氨酸‑脯氨酸。本发明通过顺式链接两种抗高血压功能肽段:IPP和VPP形成具有抗高血压能力的合成六肽,通过体内和体外生物活性验证,该抗高血压活性肽能够通过抑制ACE酶的活性,降低AngII的生成水平,从而达到降低血压的效果。本发明的活性六肽IPPVPP能够采用本领域常规方法制备,为将来药物开发中低成本和高产率的制备奠定了基础。
Description
技术领域
本发明涉及一种抗高血压活性肽,具体涉及通过将抗高血压功能肽段异亮氨酸-脯氨酸-脯氨酸(IPP)和缬氨酸-脯氨酸-脯氨酸(VPP)顺式链接并通过体外和体内生物活性实验验证的一种的抗高血压的合成六肽。
背景技术
我国高血压患病率的态势越来越严峻,当前常见的治疗高血压的药物有以下六种:利尿剂(可能引起低血钾症,影响糖代谢,使糖耐量下降)、β一阻滞剂(可带来中枢神经系统、消化系统以及血管系统的不良反应)、α一阻滞剂、钙拮抗剂(会导致水肿、头痛、潮红、多尿、低血压以及心脏传导受阻)、血管紧张素转换酶抑制剂(ACEI)和血管紧张素II受体阻滞剂(可引起干咳、血钾高、甚至血管性水肿)。由于现有大部分药物都有较强的副作用,所以科学家们一直致力于开发高效低毒的药品或食品,譬若说,含有抑制血管紧张素转换酶(angiotensinconvertingenzyme,ACE,EC3.4.15.1)活性的小肽的功能食品或药品,使高血压患者得益,本发明就是基于此背景诞生的。ACE酶是一种外肽酶,是调节血压的重要因子,主要存在于血管的内皮细胞,在肺毛中含量最为丰富。根据文献:赵海珍,陆兆新,刘战民等.天然食品来源的血管紧张素转换酶抑制肽的研究进展[J].中国生化药物杂志,2004,25(5):315-317及文献:陈强.血管紧张素转换酶抑制剂和血管紧张素II受体拮抗剂联合应用治疗肾小球疾病[J].肾脏病与透析肾移植杂志,2001,10(3):274-277的报道,ACE酶的主要作用有两个,一个是使得血管紧张素I(AngI)转化为血管紧张素II(AngII),另外一个是使缓激肽失活,以这两个作用影响肾素一血管紧张素系统(RAS)来完成血压的调节。血管紧张素II可以与血管组织中的特异性受体ATl相结合,通过血流动力学作用,使得血管收缩,收缩肾小球出球小动脉,使肾血流量下降,肾小球毛细血管内压升高,缓激肽可以通过血流动力学作用,使血管舒张,舒张肾小球出球小动脉,使肾血流量上升,肾小球毛细血管内压降低。
发明内容
本发明的目的是,提供一种高效低毒的通过抑制ACE酶活性来实现抗高血压作用的活性肽。本发明的目的还在于提供活性肽IPPVPP在制备预防或治疗高血压的药物或食物中的应用。本发明目的还提供一种抗高血压的组合物,其中含有活性肽IPPVPP作为有效成分,并含有常规药用载体和/或赋形剂。
为达到上述目的,本发明采用的技术方案是:一种抗高血压活性肽,其氨基酸序列为:异亮氨酸(Ile)-脯氨酸(Pro)-脯氨酸(Pro)-缬氨酸(Val)-脯氨酸(Pro)-脯氨酸(Pro)(简称IPPVPP)。本发明的活性肽IPPVPP选择了抗高血压功能肽段:IPP和VPP,通过顺式链接形成具有抗高血压能力的合成六肽。并采用了固相制备方法。
本发明有益效果:本发明六肽IPPVPP是基于已经得到广泛应用的两种三肽IPP和VPP顺式链接成的,拥有较好的抗高血压能力,由体外和体内生物活性进行验证。本发明获得的具有抗高血压能力的合成六肽,能够用于抑制血管紧张素转化酶(ACE酶)的活性,降低血管紧张素II(AngII)的生成水平,达到降低血压的效果。该抗高血压活性肽具有高效低毒的特点,对于开发治疗高血压药物和功能食品有着较好的应用前景。由于能通过人工化学合成技术进行大规模制备,为将来药物开发中低成本和高产率的制备奠定了基础。
附图说明
图1为大鼠喂养异亮氨酸(Ile)-脯氨酸(Pro)-脯氨酸(Pro)(简称IPP)(1.5mg/kg)后收缩压(SystolicBloodPressure,SBP)与时间的关系图;
图2为大鼠喂养IPPVPP(1.5mg/kg)后收缩压(SystolicBloodPressure,SBP)与时间的关系图;
图3为空白组大鼠收缩压(SystolicBloodPressure,SBP)与时间的关系图。
具体实施方式
本发明的活性六肽IPPVPP能够采用本领域常规方法制备,如采用固相多肽合成方法合成,具体包括以下具体步骤:
首先在脯氨酸二氯树脂上接入芴甲氧羰基脯氨酸,同时加入TBTU和DIEA进行偶联,偶联时间为30分钟,并用体积比为20%的哌啶/二甲基甲酰胺洗涤3次去除末端芴甲氧羰基基团,然后用六氢吡啶进行脱保护,脱保护结束后再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次。抽干后加入芴甲氧羰基异亮氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干。抽干后加入芴甲氧羰基脯氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干。抽干后加入芴甲氧羰基脯氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干。抽干后加入芴甲氧羰基缬氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干,最后通过半制备型反相高效液相色谱(反相柱:VYDAC-C18柱(4.6X250mm,5ym);线性梯度:溶液A(溶解于分析纯级乙腈的0.1%(v/v)三氟乙酸),梯度:5%-30%(ν/ν)25分钟、100%(v/v)25.1分钟、停止30分钟;溶液Β(溶解于纯水的0.1%(v/v)三氟乙酸),梯度:95%-70%(v/v)25分钟、0%25.1分钟、停止30分钟;流速:1.0毫升/分钟;分离收集洗脱峰,冻干后备用。
具体实施例在下面进一步阐述。六肽分子IPPVPP抗高血压活性测试:实验材料:
新西兰大白兔:上海交通大学药学院动物实验中心提供;20周龄雄性高血压模式大鼠(SHR):上海实验动物中心提供;IPP、IPPVPP,吉尔生化(上海)有限公司制备;微量移液器(100~1000μL,20~200μL,10~100μL,0.5~10μL),EppendorfLtd;过滤器(Φ50mm),上海医药工业研究所;微孔滤膜(Φ50mm,孔径为0.22μm),上海摩速科学器材有限公司;Centrifuge5415D小型高速离心机,EppendorfLtd;AKTApurifier10层析系统,Superdex30prepgrade层析柱(GEHealthcare,USA);ZorbaxbB_Cl8反相色谱柱(Agilent);摩尔超纯水机,上海摩勒科学仪器有限公司;GL-22M高速冷冻离心机,上海卢湘仪离心机仪器厂JY2002型电子天平,上海良平仪器仪表有限公司;手提式压力蒸汽灭菌器,上海医用核子仪器厂;HWS26型电热恒温水浴锅,上海一恒科技有限公司;LabDancer试管振荡器,IKAWorksGuangzhou;ElectroluxBCD-252T型冰箱,伊莱克斯(中国)电器有限公司;DW-HW138型超低温冰箱,中科美菱低温科技有限责任公司;分析天平,Meitele1-tolido,German;大鼠血压计:SoftronBP_98A,北京软隆生物技术有限公司。
合成六肽IPPVPP的体外ACE酶抑制活性结果分析:
马脲酰组氨酰亮氨酸(hippuryl-L-histidyl-L_leucine,HHL)在ACE酶的催化下快速分解产生马尿酸(HippuricAcid,HA)和二肽histidyl-leucine(HL),加入ACE酶抑制剂后,ACE酶的活性受到抑制,HA和HL的生成量减少,通过RP-HPLC方法(色谱柱=ZorbaxSB-C18柱(80A,5μm,4.6mmX250mm;流动相:20%(v/v)分析纯乙腈+0.1%(v/v)三氟乙酸;流速:1.0毫升/分钟))测定228nm下HA的生成量,按下列公式评价ACE酶抑制剂对ACE酶的抑制率。
ACE酶抑制率(%)=IOOX(A对照一A样品)/(A对照一A空白)
A对照:100mM硼酸钠缓冲液(BBS,pH8.3)代替六肽时生成HA的峰面积;A样品:六肽样品时生成HA的峰面积;A空白,盐酸先于HHL加入时IOOmM硼酸钠缓冲液(BBS,pH8.3)代替六肽时生成HA的峰面积。反应体系和条件见表1:
表1
本实施例中的ACE酶通过兔肺提取,经过酶活力测定,证明其酶活力为0.732mU/mL(—个酶单位(U)定义为在本实验条件下,每min催化三肽底物(HHL)生成IμmoI产物HA时所需的酶量),满足实验要求。于是对合成的IPPVPP以及已经得到广泛应用的两种三肽异亮氨酸(Ile)-脯氨酸(Pro)-脯氨酸(Pro)(简称IPP)、缬氨酸(Val)-脯氨酸(Pro)-脯氨酸(Pro)(简称VPP)进行试验,结果见表2,从结果可以看出,实验测得的抗高血压小肽在小肽浓度为10μM时的平均抑制率的相对大小为IPP>VPP>IPPVPP。上述结果表明本发明多肽IPPVPP对ACE酶有抑制作用,进而也说明本发明多肽IPPVPP有降低血压的功效。
表2
六肽IPPVPP的体内生物活性结果分析:
肽在体内与ACE酶反应前,还要经过消化和吸收的作用,而且在体内的理化环境十分得复杂,在体外进行的模拟结果也许不够精确。因此我们通过对自发性高血压大鼠(SpontaneouslyHypertensiveRatSHR)饲喂生物活性肽后血压的变化来衡量六肽的抗高血压效果。根据体外的活性数据的特点,选择了阳性对照IPP和IPPVPP进行体内的生物活性验证实验,表3中为大鼠给药类别:
表3
图1、2为大鼠(体重均为400g)灌胃后的血压变化情况,由试验结果可以看出,SHR高血压大鼠在喂食了二种抗高血压活性肽后,血压均发生了下降。大鼠喂养IPP(1.5mg/kg体重)后,大鼠的血压有明显的下降波谷,血压初始为195左右的正常高血压水平,在喂食IPP后1小时左右后开始下降,给药后3小时左右,下降到最低160~165,给药6小时后又重新恢复到了195~200左右的正常高血压水平,如图1所示。大鼠喂养IPPVPP(1.5mg/kg体重)后,血压值也出现了明显的波谷,药物的有效时间大约为3.5小时,血压在0.5h后开始下降,从200左右的高血压水平下降到160左右,最低点出现在给药3小时后,给药3.5小时后又恢复到初始的200水平,如图2所示。空白组大鼠饲喂了4mL去离子水后,虽然得到的数据不是十分的理想,数据的标准差略大,但是可以明显判断出,血压没有明显的降低,如图3所示。
Claims (4)
1.一种抗高血压活性肽,其特征在于:所述活性肽的氨基酸序列为:异亮氨酸-脯氨酸-脯氨酸-缬氨酸-脯氨酸-脯氨酸。
2.如权利要求1所述的抗高血压活性肽在制备预防或治疗高血压的药物或食物中的应用。
3.一种抗高血压的组合物,其特征是以异亮氨酸-脯氨酸-脯氨酸-缬氨酸-脯氨酸-脯氨酸活性肽作为有效成分,并含有常规药用载体和/或赋形剂制备成口服药物。
4.如权利要求1所述的抗高血压活性肽的制备方法,其特征在于:采用固相多肽合成方法合成,包括以下具体步骤:
首先在脯氨酸二氯树脂上接入芴甲氧羰基脯氨酸,同时加入TBTU和DIEA进行偶联,偶联时间为30分钟,并用体积比为20%的哌啶/二甲基甲酰胺洗涤3次去除末端芴甲氧羰基基团,然后用六氢吡啶进行脱保护,脱保护结束后再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次;抽干后加入芴甲氧羰基异亮氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干;抽干后加入芴甲氧羰基脯氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干;抽干后加入芴甲氧羰基脯氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干;抽干后加入芴甲氧羰基缬氨酸,TBTU和DIEA进行偶联,偶联时间为30分钟,并用20%的哌啶/二甲基甲酰胺洗涤3次,用六氢吡啶进行脱保护,再用体积比为20%的哌啶/二甲基甲酰胺洗涤6次抽干,最后通过半制备型反相高效液相色谱;线性梯度:溶液A溶解于分析纯级乙腈的0.1%(v/v)三氟乙酸,梯度:5%-30%(ν/ν)25分钟、100%(v/v)25.1分钟、停止30分钟;溶液Β溶解于纯水的0.1%(v/v)三氟乙酸,梯度:95%-70%(v/v)25分钟、0%25.1分钟、停止30分钟;流速:1.0毫升/分钟;分离收集洗脱峰,冻干得成品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610225861.9A CN105777866A (zh) | 2016-04-12 | 2016-04-12 | 一种抗高血压活性肽及制备 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610225861.9A CN105777866A (zh) | 2016-04-12 | 2016-04-12 | 一种抗高血压活性肽及制备 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN105777866A true CN105777866A (zh) | 2016-07-20 |
Family
ID=56397251
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610225861.9A Pending CN105777866A (zh) | 2016-04-12 | 2016-04-12 | 一种抗高血压活性肽及制备 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN105777866A (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107325153A (zh) * | 2017-07-26 | 2017-11-07 | 盐城卫生职业技术学院 | 一种抗高血压活性肽Citn‑Hyp‑Pro及应用和药物组合物 |
CN107337712A (zh) * | 2017-07-26 | 2017-11-10 | 盐城卫生职业技术学院 | 一种抗高血压活性肽Orn‑Hyp‑Pro及应用和药物组合物 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102786580A (zh) * | 2012-07-17 | 2012-11-21 | 上海交通大学 | 抗高血压活性肽ivp |
CN103204909A (zh) * | 2013-04-17 | 2013-07-17 | 苏州凯祥生物科技有限公司 | 一种抗高血压活性肽vppipp |
CN105001139A (zh) * | 2015-07-08 | 2015-10-28 | 南京葆赫生物技术有限公司 | 一种抗高血压活性肽、其制备方法及应用 |
-
2016
- 2016-04-12 CN CN201610225861.9A patent/CN105777866A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102786580A (zh) * | 2012-07-17 | 2012-11-21 | 上海交通大学 | 抗高血压活性肽ivp |
CN103204909A (zh) * | 2013-04-17 | 2013-07-17 | 苏州凯祥生物科技有限公司 | 一种抗高血压活性肽vppipp |
CN105001139A (zh) * | 2015-07-08 | 2015-10-28 | 南京葆赫生物技术有限公司 | 一种抗高血压活性肽、其制备方法及应用 |
Non-Patent Citations (1)
Title |
---|
郭慧青等: "两种血管紧张素转化酶抑制肽作用于靶标的分子机理", 《食品科学》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107325153A (zh) * | 2017-07-26 | 2017-11-07 | 盐城卫生职业技术学院 | 一种抗高血压活性肽Citn‑Hyp‑Pro及应用和药物组合物 |
CN107337712A (zh) * | 2017-07-26 | 2017-11-10 | 盐城卫生职业技术学院 | 一种抗高血压活性肽Orn‑Hyp‑Pro及应用和药物组合物 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Nurminen et al. | α-Lactorphin lowers blood pressure measured by radiotelemetry in normotensive and spontaneously hypertensive rats | |
Suat et al. | Design of β-turn based therapeutic agents | |
EP3145943B1 (en) | Peptides, compositions comprising them and uses in particular cosmetic uses | |
US9266921B2 (en) | PGC-1α-modulating peptides | |
Saito et al. | Structure and activity of angiotensin I converting enzyme inhibitory peptides from sake and sake lees | |
Ung et al. | Tripeptide motifs in biology: targets for peptidomimetic design | |
TW496874B (en) | 5-Membered ring heterocycles as inhibitors of leucocyte adhesion and as VLA-4 antagonists | |
CN101153055B (zh) | 具有血管紧张素转移酶抑制活性的新型肽及其制备方法 | |
CN105017122B (zh) | 一种抗高血压活性肽、其制备方法及应用 | |
CN105001139B (zh) | 一种抗高血压活性肽、其制备方法及应用 | |
CN102786579B (zh) | 抗高血压活性肽vip | |
US20230190616A1 (en) | Compounds useful for the treatment and/or care of the skin, hair, nails and/or mucous membranes | |
CN101210047A (zh) | 一种活性单肽及其应用 | |
CN105777866A (zh) | 一种抗高血压活性肽及制备 | |
CN102786580A (zh) | 抗高血压活性肽ivp | |
Yu et al. | Anxiolytic effects of ACE inhibitory peptides on the behavior of rats in an elevated plus-maze | |
CN103204909B (zh) | 一种抗高血压活性肽vppipp | |
CN102399261A (zh) | 具有血管紧张素转化酶c-端选择性抑制活性的三肽及其应用和组合物 | |
CN107337712A (zh) | 一种抗高血压活性肽Orn‑Hyp‑Pro及应用和药物组合物 | |
CN101744845A (zh) | 绿藻萃取组合物及其制备方法 | |
CN113072621A (zh) | 一种牦牛骨降血压肽及其制备方法与应用 | |
CN1087283C (zh) | 作用改善的新lh-rh拮抗剂 | |
CN105153282A (zh) | 一种十肽及其应用 | |
CN112694429B (zh) | 一类多肽及其在制备ace抑制剂或降血压产品上的应用 | |
CN109265538A (zh) | 鲽鱼皮来源的活性二肽 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160720 |