CN110577574B - 1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 - Google Patents
1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 Download PDFInfo
- Publication number
- CN110577574B CN110577574B CN201810589789.7A CN201810589789A CN110577574B CN 110577574 B CN110577574 B CN 110577574B CN 201810589789 A CN201810589789 A CN 201810589789A CN 110577574 B CN110577574 B CN 110577574B
- Authority
- CN
- China
- Prior art keywords
- lys
- methyl
- obzl
- boc
- carboline
- 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 - Fee Related
Links
- 230000000694 effects Effects 0.000 title abstract description 14
- 230000015572 biosynthetic process Effects 0.000 title description 2
- 238000003786 synthesis reaction Methods 0.000 title description 2
- 238000002360 preparation method Methods 0.000 claims abstract description 29
- 208000032382 Ischaemic stroke Diseases 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 11
- 230000002785 anti-thrombosis Effects 0.000 claims abstract description 9
- 239000003146 anticoagulant agent Substances 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 15
- 238000004519 manufacturing process Methods 0.000 claims 2
- 239000003527 fibrinolytic agent Substances 0.000 claims 1
- 241000700159 Rattus Species 0.000 abstract description 41
- 150000001875 compounds Chemical class 0.000 abstract description 39
- 230000002537 thrombolytic effect Effects 0.000 abstract description 8
- 208000006011 Stroke Diseases 0.000 abstract description 5
- FYQSMXKJYTZYRP-DCAQKATOSA-N Pro-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 FYQSMXKJYTZYRP-DCAQKATOSA-N 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 36
- 239000000243 solution Substances 0.000 description 28
- 208000007536 Thrombosis Diseases 0.000 description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 19
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 210000001367 artery Anatomy 0.000 description 15
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 12
- 230000002829 reductive effect Effects 0.000 description 12
- 239000011780 sodium chloride Substances 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 10
- 239000004698 Polyethylene Substances 0.000 description 10
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 10
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 10
- -1 polyethylene Polymers 0.000 description 10
- 229920000573 polyethylene Polymers 0.000 description 10
- 239000007787 solid Substances 0.000 description 10
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 9
- 239000008280 blood Substances 0.000 description 9
- 210000004369 blood Anatomy 0.000 description 9
- 210000001168 carotid artery common Anatomy 0.000 description 9
- 239000002504 physiological saline solution Substances 0.000 description 9
- 229920006395 saturated elastomer Polymers 0.000 description 9
- 238000003756 stirring Methods 0.000 description 9
- 210000003462 vein Anatomy 0.000 description 9
- 239000007864 aqueous solution Substances 0.000 description 8
- 210000004556 brain Anatomy 0.000 description 8
- 206010008118 cerebral infarction Diseases 0.000 description 8
- 230000017531 blood circulation Effects 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 229920001971 elastomer Polymers 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- FABREHIBEDUZFU-UHFFFAOYSA-N 3-O-benzyl 2-O-tert-butyl 1-methyl-1,3,4,9-tetrahydropyrido[3,4-b]indole-2,3-dicarboxylate Chemical compound C(C1=CC=CC=C1)OC(=O)C1N(C(C=2NC3=CC=CC=C3C=2C1)C)C(=O)OC(C)(C)C FABREHIBEDUZFU-UHFFFAOYSA-N 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 5
- 210000000269 carotid artery external Anatomy 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- SOHLZANWVLCPHK-LBPRGKRZSA-N (2s)-2-[(2-methylpropan-2-yl)oxycarbonylamino]-4-oxo-4-phenylmethoxybutanoic acid Chemical compound CC(C)(C)OC(=O)N[C@H](C(O)=O)CC(=O)OCC1=CC=CC=C1 SOHLZANWVLCPHK-LBPRGKRZSA-N 0.000 description 4
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- NMTZYNFLSYSADY-UHFFFAOYSA-N benzyl 1-methyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylate Chemical compound C(C1=CC=CC=C1)OC(=O)C1NC(C=2NC3=CC=CC=C3C=2C1)C NMTZYNFLSYSADY-UHFFFAOYSA-N 0.000 description 4
- 210000004004 carotid artery internal Anatomy 0.000 description 4
- 208000026106 cerebrovascular disease Diseases 0.000 description 4
- 230000008034 disappearance Effects 0.000 description 4
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 4
- 210000004731 jugular vein Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 4
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 4
- 201000006474 Brain Ischemia Diseases 0.000 description 3
- 206010008120 Cerebral ischaemia Diseases 0.000 description 3
- 208000032843 Hemorrhage Diseases 0.000 description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 3
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 3
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 230000000740 bleeding effect Effects 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 231100000517 death Toxicity 0.000 description 3
- 229920000669 heparin Polymers 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 230000007971 neurological deficit Effects 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- CHKVPAROMQMJNQ-UHFFFAOYSA-M potassium bisulfate Chemical compound [K+].OS([O-])(=O)=O CHKVPAROMQMJNQ-UHFFFAOYSA-M 0.000 description 3
- 229910000343 potassium bisulfate Inorganic materials 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 238000012353 t test Methods 0.000 description 3
- 229960005356 urokinase Drugs 0.000 description 3
- VVQIIIAZJXTLRE-QMMMGPOBSA-N (2s)-2-amino-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound CC(C)(C)OC(=O)NCCCC[C@H](N)C(O)=O VVQIIIAZJXTLRE-QMMMGPOBSA-N 0.000 description 2
- OHVLMTFVQDZYHP-UHFFFAOYSA-N 1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-2-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]ethanone Chemical compound N1N=NC=2CN(CCC=21)C(CN1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)=O OHVLMTFVQDZYHP-UHFFFAOYSA-N 0.000 description 2
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 2
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 2
- 206010061216 Infarction Diseases 0.000 description 2
- VGALFAWDSNRXJK-VIFPVBQESA-N L-aspartic acid beta-benzyl ester Chemical compound OC(=O)[C@@H](N)CC(=O)OCC1=CC=CC=C1 VGALFAWDSNRXJK-VIFPVBQESA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 2
- 210000001715 carotid artery Anatomy 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 2
- 229960001008 heparin sodium Drugs 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- MTNDZQHUAFNZQY-UHFFFAOYSA-N imidazoline Chemical compound C1CN=CN1 MTNDZQHUAFNZQY-UHFFFAOYSA-N 0.000 description 2
- 150000002462 imidazolines Chemical class 0.000 description 2
- 230000007574 infarction Effects 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 230000007658 neurological function Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- FYQSMXKJYTZYRP-UHFFFAOYSA-N 6-amino-2-[2-(pyrrolidine-2-carbonylamino)propanoylamino]hexanoic acid Chemical compound NCCCCC(C(O)=O)NC(=O)C(C)NC(=O)C1CCCN1 FYQSMXKJYTZYRP-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- JXYACYYPACQCDM-UHFFFAOYSA-N Benzyl glycinate Chemical compound NCC(=O)OCC1=CC=CC=C1 JXYACYYPACQCDM-UHFFFAOYSA-N 0.000 description 1
- 101100510326 Caenorhabditis elegans tpa-1 gene Proteins 0.000 description 1
- 208000022540 Consciousness disease Diseases 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010037394 Pulmonary haemorrhage Diseases 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 241000221566 Ustilago Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- YIRBOOICRQFSOK-NSHDSACASA-N benzyl (2s)-2-amino-3-methylbutanoate Chemical compound CC(C)[C@H](N)C(=O)OCC1=CC=CC=C1 YIRBOOICRQFSOK-NSHDSACASA-N 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 210000003194 forelimb Anatomy 0.000 description 1
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000002636 imidazolinyl group Chemical group 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 210000004761 scalp Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002444 silanisation Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Cardiology (AREA)
- Vascular Medicine (AREA)
- Heart & Thoracic Surgery (AREA)
- Urology & Nephrology (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
技术领域
本发明公开了下式的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val,涉及它的制备方法,涉及它的抗血栓活性,涉及它的溶血栓活性以及涉及它治疗中风24小时大鼠的作用,因而本发明涉及它在制备抗血栓药物,溶血栓药物以及治疗缺血性中风药物中的应用。本发明属于生物医药领域。
技术背景
缺血性中风是一类较常见且危害严重的脑血管疾病,特点是发病率高、病死率高、致残率高和复发率高。目前临床治疗缺血性中风面临没有有效药物的现实,尤其中风面4h以上的患者非死即残。发明对中风面4h以上的患者有效的药物是临床的重要需求。发明人曾经公开下式的咪唑啉在中风面24h的大鼠缺血性中风模型上,显示优秀疗效。即连续静脉注射6天下式的咪唑啉,每天1次,剂量为100nmol/kg,具有优秀疗效。式中AA为Ser,Val或Phe。由于结构的原因,下式的咪唑啉有两个必然缺点。即1,3-二氧咪唑啉部分的自由基对还原性环境敏感,不仅制备困难,而且保存困难。
发明人在经过3年实验研究,发现咪唑啉部分用1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基替代可得到稳定结构和容易保存的双重意想不到的技术效果。按照这个发现发明人提出了本发明。
发明内容
本发明的第一个内容是提供下式的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val。
本发明的第二个内容是提供1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val的合成方法,该方法包括:
(1)制备N-Boc-1S-甲基-1,2,3,4四氢-β-咔啉-3S-羧酸;
(2)制备N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc);
(3)制备HCl·Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl;
(4)制备Boc-Pro-Ala-Lys(Cbz);
(5)制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl;
(6)制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val。
本发明的第三个内容是评价1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val的抗血栓活性,溶血栓活性以及治疗缺血性中风的活性。
附图说明
图1 1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val的合成路线。i:CH3CHO,H2O;ii:TEA,DMF,二碳酸二叔丁酯(Boc)2O;iii:Pb/C,H2,四氢呋喃;iv:N,N-二环己基碳二亚(DCC),N-羟基苯并三氮唑(HOBt),N-甲基吗啉(NMM),四氢呋喃;v:2N氢氧化钠水溶液,CH3OH;vi:氯化氢的乙酸乙酯溶液(4M);vii:三氟醋酸(TFA),三氟甲磺酸(TMFA)。
具体实施方式
为了进一步阐述本发明,下面给出一系列实施例。这些实施例完全是例证性的,它们仅用来对本发明进行具体描述,不应当理解为对本发明的限制。
实施例1制备1-甲基-1,2,3,4-四氢-β-咔啉-3-羧酸苄酯(1)
向800mL蒸馏水中逐滴加入1mL浓度为98%的浓硫酸,搅拌均匀后将10.0g(34mmol)L-Trp-OBzl分三次加入其中。搅拌五分钟,使L-Trp-OBzl与硫酸水溶液充分混悬。之后,往浮悬液中滴加入10mL浓度为40%乙醛水溶液。反应化合物搅拌12h,然后往里滴加3mL浓氨水调反应溶液pH值至8。反应化合物静置1h,待产物充分析出。滤出固体,干燥,得9.84g(90%)淡黄色固体,为1R-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯和1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯的混合物。ESI-MS(m/e):321[M+H]+。
实施例2制备N-Boc-1-甲基-1,2,3,4-四氢-β-咔啉-3-羧酸苄酯(2)
将实施例1得到的9.84g(30.8mmol)1-甲基-1,2,3,4-四氢-β-咔啉-3-羧酸苄酯用20mL N,N-二甲基甲酰胺(DMF)溶解。于0℃往溶液中加6.98g(32.0mmol)(Boc)2O。得到的溶液用三乙胺调节pH值12,室温搅拌48h。反应混合物减压浓缩,除去DMF。残留物用100mL乙酸乙酯溶解。得到的乙酸乙酯溶液依次用5%硫酸氢钾水溶液洗(50mL×3)和饱和氯化钠水溶液洗(50mL×3)。分离的乙酸乙酯层用无水硫酸钠干燥12h,过滤,滤液减压浓缩得到的油状物。该油状物用硅胶柱分离(二氯甲烷/甲醇,100/1),得到5.02g(38%)N-Boc-1R-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯和6.00g(47%)N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯。均为无色粉末。
实施例3制备Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸(3)
往1.0g(2.38mmol)N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯与40mL无水四氢呋喃的溶液中加100mg Pd/C,搅拌使成为均匀的悬浮液。减压抽出反应体系内的空气,通入氢气,室温搅拌10h,TLC(二氯甲烷/甲醇,40/1)显示N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯完全消失。过滤除去Pd/C,滤液减压浓缩,得到的无色粉末的ESI-MS(m/e)为329[M-H]-。
实施例4制备N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-酰基–Lys(Boc)-OBzl(4)
将4.950g(15.0mmol)Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3-羧酸溶于50mL无水四氢呋喃,冰浴与搅拌下依次往得到的溶液中加入2.025g(15.0mmol)N-羟基苯骈三氮唑(HOBt)和用无水THF溶解的3.090g(15.0mmol)N,N-二环己基碳二亚胺(DCC)溶液,充分搅拌活化30分钟得溶液A;然后将用20ml无水THF溶解的5.066g(13.6mmol)HCl·Lys(Boc)-OBzl用N-甲基吗啉先调节pH值至8-9。再加入到反应液A中。撤去冰浴室温搅拌8小时,UV(254nm)下TLC(展开剂二氯甲烷/甲醇=40/1)显示原料HCl·Lys(Boc)-OBzl完全消失。滤液减压浓缩,残留物用乙酸乙酯溶解。乙酸乙酯层依次用饱和碳酸氢钠水溶液萃洗(n=3)、饱和氯化钠水溶液萃洗(n=3)、5%硫酸氢钾水溶液萃洗(n=3)、饱和氯化钠水溶液萃洗(n=3)、饱和碳酸氢钠水溶液萃洗(n=3)和饱和氯化钠水溶液萃洗(n=3),合并上层乙酸乙酯相,用无水硫酸钠干燥4小时,过滤,得到的滤液减压浓缩至干,经硅胶柱层析,得到6.960g(79%)标题化合物,为无色粉末。ESI-MS(m/e):649[M+H]+。
实施例5制备N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc)(5)
采用实施例3的方法从1.296g(2.0mmol)N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-羧酸苄酯得到标题化合物,为无色粉末。ESI-MS(m/e):557[M-H]-。
实施例6制备Boc-Arg(NO2)-Gly-OBzl
采用实施例4的方法从5.000g(15.6mmol)Boc-Arg(NO2)和2.860g(14.2mmol)HCl·Gly-OBzl得到5.560g(84%)标题化合物,为无色粉末。ESI-MS(m/e):467[M+H]+。
实施例7制备Boc-Arg(NO2)-Gly
将4.660g(1mmol)Boc-Arg(NO2)-Gly-OBzl溶于50mL甲醇。冰浴与搅拌下往得到的溶液中缓慢滴加2N NaOH水溶液,冰浴搅拌6小时。UV(254nm)下TLC(展开剂二氯甲烷/甲醇=40/1)显示Boc-Arg(NO2)-Gly-OBzl完全消失。冰浴与搅拌下缓慢滴加饱和KHSO4水溶液调pH至7,减压浓缩除去甲醇。用饱和硫酸氢钾水溶液调pH为2,再用乙酸乙酯(40mL×3)反复萃取水溶液3遍,饱和氯化钠水溶液萃洗3次,合并乙酸乙酯层,用无水硫酸钠干燥2小时,过滤,滤液减压浓缩至干,得到2.850g(76%)标题化合物。ESI-MS(m/e):375[M-H]-。
实施例8制备Boc-Asp(OBzl)-Val-OBzl
采用实施例4的方法从3.230g(10mmol)Boc-Asp(OBzl)和3.450g(9.1mmol)Tos.Val-OBzl得到3.910g(84%)标题化合物,为无色固体。ESI-MS(m/e):515[M+H]+。
实施例9制备Asp(OBzl)-Val-OBzl
将10.0g Boc-Asp(OBzl)-Val-OBzl用10mL无水乙酸乙酯溶解。在冰浴与搅拌下往该溶液中加入100mL氯化氢的乙酸乙酯溶液(4M);搅拌2h。UV(254nm)下TLC(二氯甲烷/甲醇,40/1)显示原料Boc-Asp(OBzl)-Val-OBzl完全消失。反应混合物减压浓缩至干,残留物用30mL无水乙酸乙酯稀释,再减压浓缩至干。该操作重复3次。得到的残留物用30mL无水乙醚稀释,再减压浓缩至干。该操作重复3次,得到标题化合物。ESI-MS(m/e):413[M+H]+。
实施例10制备Boc-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl
采用实施例4的方法从3.750g(10.0mmol)Boc-Arg(NO2)-Gly和4.080g(9.1mmol)HCl·Asp(OBzl)-Val-OBzl得到5.190g(74%)标题化合物,为无色固体。ESI-MS(m/e):770[M+H]+。
实施例11制备Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl
采用实施例9的方法从5.0g(6.5mmol)Boc-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl得到4.27g(98%)标题化合物,为无色固体。ESI-MS(m/e):673[M+H]+。
实施例12制备Boc-Pro-Ala-OBzl
采用实施例4的方法从2.150g(10mmol)Boc-Pro和1.960g(9.1mmol)HCl·Ala-OBzl得到3.040g(90%)标题化合物,为无色固体。ESI-MS(m/e):377[M+H]+
实施例13制备Boc-Pro-Ala
采用实施例3的方法从1.880g(5mmol)Boc-Pro-Ala-OBzl得到得到1.350g(95%)标题化合物,为无色固体。ESI-MS(m/e):285[M-H]+。
实施例14制备Boc-Pro-Ala-Lys(Cbz)-OBzl
采用实施例4的方法从10.0g Boc-Pro-Ala-OBzl和14.2g HCl·Lys(Cbz)-OBzl得到14.5
g(65%)标题化合物,为无色油状产物。ESI-MS(m/e):639[M+H]+。
实施例15制备Boc-Pro-Ala-Lys(Cbz)
采用实施例7的方法从1.280g(2.0mmol)Boc-Pro-Ala-Lys(Cbz)-OBzl,得到0.876g(79%)标题化合物,为无色油状物。ESI-MS(m/e):549[M+H]+。
实施例16制备N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc)-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl(6)
采用实施例4的方法从5.580g(10.0mmol)N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc)和6.430g(9.1mmol)HCl·Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl得到8.148g(74%)标题化合物,为无色固体。ESI-MS(m/e):1211[M+H]+。
实施例17制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl(7)
采用实施例9的方法1.210g(1.0mmol)N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc)-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl得到0.96g(95%)标题化合物。ESI-MS(m/e):1011[M+H]+。
实施例18制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys[Boc-Pro-Ala-Lys(Cbz)]-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl(8)
采用实施例4的方法从1.096g(2.0mmol)Boc-Pro-Ala-Lys(Cbz)和1.840g(1.82mmol)1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl得到415mg(15%)标题化合物,为无色固体。ESI-MS(m/e):1541[M+H]+。1H NMR(DMSO-d6,300MHz)δ/ppm=10.77(s,1H),8.53(s,1H),8.17(m,4H),8.04(m,2H),7.86(m,2H),7.71(m,2H),7.34(m,15H),7.30(m,2H),7.19(t,J=5.1Hz,1H),7.01(m,2H),5.08(m,6H),4.79(m,1H),4.31(m,3H),4.16(m,4H),3.74(d,J=4.8Hz,1H),3.51(m,1H),3.16(m,3H),2.96(m,5H),2.72(m,2H),2.05(m,2H),1.74(m,5H),1.55(m,5H),1.38(m,19H),1.20(m,6H),0.84(d,J=6.3Hz,6H)。
实施例19制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Arg-Pro-Ala-Lys)-Arg-Gly-Asp-Val-OH(9)
将100.0mg(0.065mmol)1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys[Boc-Pro-Ala-Lys(Cbz)]-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl在冰盐浴下加入1mL三氟醋酸(TFA)使其完全溶解,再加入0.30mL三氟甲磺酸(TFMSA),1小时后UV(254nm)下TLC(展开剂正丁醇:水:冰醋酸=1:1:1)显示原料点消失,停止反应,向反应中加入无水乙醚搅拌,固体析出,静置,弃去上清夜,再加入无水乙醚,静置,弃去上清夜,重复3次,减压抽干,然后用少量水溶解,用25%氨水调pH至7,用Sephadex G15除盐,C18柱纯化,冻干,得到10mg(15%)标题化合物,为无色粉末。ESI-MS(m/e):1082[M+H]+;mp:210.3-211.1℃;IR(cm-1):3264.58,3058.83,2935.76,2864.95,1644.95,1524.97,1454.29,1391.15,1245.08,1099.16,743.60,649.07;1H NMR(DMSO-d6,300MHz)δ/ppm=10.80(s,1H),10.26(s,1H),8.89(m,2H),8.55(s,1H),8.35(m,2H),8.06(m,2H),7.38(d,J=7.5Hz,1H),7.29(d,J=7.5Hz,1H),7.04(m,5H),4.53(m,2H),4.29(m,3H),4.16(m,2H),3.92(m,5H),3.03(m,6H),2.75(m,3H),2.63(m,2H),2.01(m,3H),1.75(m,5H),1.53(m,7H),1.46(m,4H),1.26(m,9H)0.79(d,J=6.6Hz,6H)。
实验例1评价化合物9的抗血栓活性
将雄性SD大鼠(200±20g),随机分组,每组10只,饲养1天,停止喂食过夜。灌胃给予化合物9的生理盐水溶液(剂量10nmol/kg)或阿司匹林的生理盐水溶液(剂量167μmol/kg)或生理盐水(剂量3mL/kg)30min之后,大鼠用20%乌来糖的生理盐水溶液麻醉,之后手术。分离大鼠的右颈动脉和左颈静脉,将准确称重的丝线置于旁路插管,管的一端插入左静脉,另一端管插入右侧动脉并注射0.2mL肝素钠抗凝。使得血流从右侧动脉流经旁路插管进入左侧静脉,15min之后取出附有血栓的丝线称量,计算血液循环前后丝线的重量,得到的血栓重以均值±SD mg表示并代表抗血栓活性,作t检验。数据列入表1。结果表明口服10nmol/kg化合物9能有效地抑制血栓形成。
表1在10nmol/kg剂量下化合物9的抗血栓活性
a)与生理盐水比P<0.01,n=10
实验例2评价化合物9的溶血栓活性
SD大鼠(雄性,200±20g)按1200mg/kg的剂量腹腔注射乌拉坦生理盐水溶液进行麻醉。麻醉大鼠后将其仰卧位固定,分离其右颈总动脉,近心端处夹住动脉夹,将近心端及远心端分别穿入手术线,远心端的手术线结扎,远心端插管,将动脉夹松开,取出1mL动脉血,置于1mL离心管中。往垂直固定的橡胶管(长5mm,内径2.5mm,外径5.0mm,管底用胶塞密封,para膜封紧)内注入0.1mL大鼠动脉血,随后在管内迅速插入一支不锈钢材质的血栓的固定螺栓(血栓固定螺旋用直径为0.2mm的不锈钢丝绕成,螺旋部分长10mm,内含15个螺圈,螺圈的直径为1.0mm,托柄与螺旋相连,长约7.0mm,呈问号型)。
旁路插管由三部分构成,中间段为长60.0mm,内径3.5mm的聚乙烯胶管;两端均为长100.0mm,内径1.0mm,外径2.0mm的相同的聚乙烯管,该管一端拉成尖管,长约10.0mm(用于插入大鼠颈动脉及静脉),外径为1.0mm,其另一端的外部套一段长为7.0mm,外径为3.5mm的聚乙烯管(用于插入中段的聚乙烯胶管内),3段管的内壁均需要硅烷化(1%的硅油乙醚溶液)。将血栓包裹的血栓固定螺旋置于中段聚乙烯胶管内,胶管的另外两端分别与两根聚乙烯的加粗端相套,保证在循环的过程中不会漏血。用注射器通过尖管端将管中注满肝素生理盐水溶液(50IU/kg),排除气泡,备用。
分离大鼠的左颈外静脉,近心端和远心端分别穿入手术线,结扎远心端的血管,在暴露的左颈外静脉上剪一小口,将上述制备好的旁路管道尖管由小口插入左颈外静脉开口处,同时远离旁路管中段(含精确称量的血栓固定螺旋)内血栓固定螺旋。用注射器通过另一端的尖管注入准确量的肝素钠的生理盐水溶液(50IU/kg),此时注射器不要撤离聚乙烯管,用动脉夹夹住注射器与聚乙烯管之间的软管。在右颈总动脉的近心端用动脉夹止血,结扎远心端,在离动脉夹不远处将右颈总动脉剪一小口,从聚乙烯管的尖部拔出注射器,将聚乙烯管的尖部插入动脉斜口的近心端。旁路管道的两端均用4号手术缝线将动静脉固定。
用头皮针将生理盐水(3mL/kg)或尿激酶的生理盐水溶液(剂量为20000IU/kg)或化合物9的生理盐水溶液(剂量为10nmol/kg)通过旁路管的中段(含精确称量的血栓固定螺旋),扎入远离血栓固定螺旋的近静脉端,松开动脉夹,使血流通过旁路管道从动脉流向静脉。将注射器中的溶液缓慢注入血液,通过血液循环,按静脉-心脏-动脉的顺序作用于螺旋的血栓上。血液循环1h之后,从旁路管道中取出固定血栓的螺旋,精确称量。计算每只大鼠旁路管道中固定血栓的螺旋血液循环前后血栓的重量差,即血栓减重。对数据(均值±SDmg)作t检验。血栓减重代表溶血栓活性。表2的结果表明,10nmol/kg化合物9能有效地溶解血栓。活性与20000IU/kg尿激酶无显著差异。说明技术效果明显。
表2在10nmol/kg剂量下化合物9的溶血栓活性
a)与生理盐水比P<0.01,与尿激酶比P>0.05;n=9.
实验例3评价化合物9对缺血性中风24h大鼠的治疗作用
在雄性SD大鼠(体重300±20g)的颈部正中部竖直开约2cm长切口,沿胸锁乳突肌内侧缘分离出右颈总动脉、颈外动脉及颈内动脉。用无创动脉夹分别夹闭颈内动脉开口处和颈总动脉近心端,结扎颈外动脉的远心端,在颈外动脉剪一小口,松开颈总动脉近心端的动脉夹,取10μL血,之后再用无创动脉夹夹闭颈总动脉的近心端。将取得的10μL血放置在1mLEP管中常温放置30分钟使血液凝固,然后转移至-20℃冰箱中放置1小时,使血液凝块结实。大鼠用10%水合氯醛腹腔注射麻醉,剂量为400mg/kg。取出血液凝块,加入1mL生理盐水,用钢铲把血液凝块捣成大小均一的细小血栓块,制备细小血栓的悬液并转移至1mL注射器内。松开颈总动脉近心端的动脉夹,将1mL血栓混悬液缓慢从大鼠颈外动脉向近心端经过颈内动脉注入大鼠的大脑,然后结扎颈外动脉近心端,打开颈内动脉和颈总动脉处得动脉夹,恢复血流。等待苏醒。大鼠苏醒24小时后按Zealonga方法评定神经功能缺损程度。0分表示无任何神经功能缺失体征、1分表示未损伤侧前肢不能伸展、2分表示向未损伤侧行走、3分表示向未损伤侧转圈成追尾状行走、4分表示意识障碍无自主行走、5分表示死亡。按照得分平均分组。大鼠每天经尾静脉注射1次生理盐水,剂量为3mL/kg。连续注射6天,每天评分。表3的数据说明,生理盐水给药治疗前大鼠的神经生物学评分为5只2分,4只3分,2只4分,生理盐水给药治疗6天后大鼠的神经生物学评分为5只死亡,1只0分,2只1分,3只2分。整体表现呈现出病情恶化。
表3生理盐水连续治疗6天对脑缺血24小时大鼠神经生物学评分的影响
剂量:3mL/kg;n=11
大鼠每天经尾静脉注射1次tPA,剂量为3mg/kg。每天评分。表4的数据说明,6只大鼠有两只于24小时内死亡,对死亡大鼠进行了尸体解剖,发现它们的脏器都有出血现象,尤其是肺出血严重,于是,在给药2次之后终止给药。2次给药没有大鼠好转为无任何神经功能缺失体征,有2只残留轻微的神经功能缺失体征,有1只残留未损伤侧转圈成追尾状行走体征,有一只残留意识障碍无自主行走体征。
表4大鼠缺血6小时后接受3mg/kg tPA治疗的疗效
剂量:3mg/kg;n=6
大鼠每天经尾静脉注射1次化合物9,剂量为10nmol/kg。连续注射6天,每天评分。表5的数据说明,化合物9治疗前大鼠的神经生物学评分为1只1分,4只2分,4只3分,2只4分,给药治疗6天后大鼠的神经生物学评分为1只死亡,5只0分,5只1分。此外,存活的大鼠均未见眼眶和尾巴出血。整体显示疗效明显。
表5化合物9连续治疗6天对脑缺血24小时大鼠神经生物学评分的影响
剂量:100nmol/kg;n=11
实验例4评价化合物9对缺血性中风24h大鼠脑梗死体积的影响
接受实验例3评价神经功能缺损程度后,用乌拉坦麻醉,迅速断头取脑置于-20℃冰箱冻2小时,从前额极开始连续切片,制得6片2mm厚冠状脑切片。将脑切片置于2%TTC溶液中37℃避光孵育30min,观察脑切片的颜色变化。待正常脑组织被TTC染成红色,缺血脑组织呈白色后,用数码相机照相。照片经SPSS统计软件处理,计算脑冠状切片中梗死体积和正常组织的体积,统计各组的梗死体积百分比值。实验数据均采用t检验。表6的数据说明,化合物9治疗的缺血性中风24h大鼠脑梗死体积显著小于生理盐水治疗的缺血性中风24h大鼠脑梗死体积。
表6化合物9连续治疗6天对脑缺血24小时大鼠的脑梗死体积比
a)数据来自6/11只存活的大鼠脑切片;b)数据来自10/11只存活的大鼠脑切片,与生
理盐水比P<0.01;n=11
总结:与曾经公开的化合物相比,本发明的化合物稳定容易制备和保存。一次性口服10nmol/kg化合物9可有效地抑制血栓形成,一次性注射10nmol/kg化合物9可有效地溶解血栓。连续6天每天一次静脉给予100nmol/kg化合物9可有效地恢复缺血性中风24小时大鼠的神经生物学行为,可有效地减小缺血性中风24小时大鼠的脑梗死体积。也就是说,本发明获得了突出的技术效果。
Claims (5)
2.权利要求1的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val的制备方法,该方法包括:
(1)制备N-Boc-1S-甲基-1,2,3,4四氢-β-咔啉-3S-羧酸;
(2)制备N-Boc-1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Boc);
(3)制备HCl·Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl;
(4)制备Boc-Pro-Ala-Lys(Cbz);
(5)制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl;
(6)制备1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基–Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val。
3.权利要求1的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val在制备抗血栓药物中的应用。
4.权利要求1的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val在制备溶血栓药物中的应用。
5.权利要求1的1S-甲基-1,2,3,4-四氢-β-咔啉-3S-甲酰基-Lys(Pro-Ala-Lys)-Arg-Gly-Asp-Val在制备治疗缺血性中风药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810589789.7A CN110577574B (zh) | 2018-06-08 | 2018-06-08 | 1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810589789.7A CN110577574B (zh) | 2018-06-08 | 2018-06-08 | 1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110577574A CN110577574A (zh) | 2019-12-17 |
CN110577574B true CN110577574B (zh) | 2022-08-09 |
Family
ID=68808931
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810589789.7A Expired - Fee Related CN110577574B (zh) | 2018-06-08 | 2018-06-08 | 1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110577574B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106608905A (zh) * | 2015-10-22 | 2017-05-03 | 彭莉 | 四氢异喹啉-3-甲酰-k(grpak)rgdv,其合成,活性及应用 |
CN107686509A (zh) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | 甲基‑β‑咔啉‑3‑甲酰RGD肽,其合成,抗血栓作用和应用 |
-
2018
- 2018-06-08 CN CN201810589789.7A patent/CN110577574B/zh not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106608905A (zh) * | 2015-10-22 | 2017-05-03 | 彭莉 | 四氢异喹啉-3-甲酰-k(grpak)rgdv,其合成,活性及应用 |
CN107686509A (zh) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | 甲基‑β‑咔啉‑3‑甲酰RGD肽,其合成,抗血栓作用和应用 |
Non-Patent Citations (3)
Title |
---|
A class of oral N-[(1S,3S)-1-methyl-1,2,3,4-tetrahydro-b-carboline-3-carbonyl]-N0-(amino-acid-acyl)hydrazine: Discovery, synthesis, in vitro anti-platelet aggregation/in vivo anti-thrombotic evaluation and 3D QSAR analysis;Yao et al.;《European Journal of Medicinal Chemistry》;20110429;全文 * |
DHDMIQK(KAP): a novel nano-delivery system of dihydroxyl-tetrahydro-isoquinoline-3-carboxylic acid and KPAK towards the thrombus;Feng et al.;《Journal of Materials Chemistry B》;20161231;全文 * |
Nanosized Aspirin-Arg-Gly-Asp-Val: Delivery of Aspirin to Thrombus by the Target Carrier Arg-Gly-Asp-Val Tetrapeptide;Jin et al.;《ACS Nano》;20130812;第07卷(第09期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN110577574A (zh) | 2019-12-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108948146B (zh) | 1R-甲基-β-四氢咔啉酰-K(ARPAK)-RGDV,其合成,活性和应用 | |
CN108929372B (zh) | 1R-甲基-β-四氢咔啉酰-K(GRPAK)-RGDV,其合成,活性和应用 | |
CN110577572B (zh) | 1S-甲基-β-四氢咔啉酰-K(PAK)其合成,活性和应用 | |
CN110577574B (zh) | 1S-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 | |
CN108948155B (zh) | 1R-甲基-β-四氢咔啉酰-K(QRPAK)-RGDV,其合成,活性和应用 | |
CN108948145B (zh) | 1R-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 | |
CN110577575B (zh) | 1S-甲基-β-四氢咔啉酰-K(ARPAK)-RGDV,其合成,活性和应用 | |
CN106608905B (zh) | 四氢异喹啉-3-甲酰-k(grpak)rgdv,其合成,活性及应用 | |
CN110615828B (zh) | 1S-甲基-β-四氢咔啉酰-K其合成,活性和应用 | |
CN106432417B (zh) | 五甲氧色胺基羰丙酰-rpak,其制备,活性和应用 | |
CN110577578B (zh) | 1S-甲基-β-四氢咔啉酰-K(RPAK)其合成,活性和应用 | |
CN106317196B (zh) | 咪唑并吡啶甲酰-k(k)-aa1-aa2-aa3-ak,其合成,活性及应用 | |
CN111848727B (zh) | 茶氨酰四氢咪唑并吡啶-6-甲酰非极性氨基酸的制备,活性和应用 | |
CN110577584A (zh) | 1S-甲基-β-四氢咔啉酰-K(RPAK)-RGDV,其合成,活性和应用 | |
CN110577576A (zh) | 1S-甲基-β-四氢咔啉酰-K(GRPAK)-RGDV,其合成,活性和应用 | |
CN106608901B (zh) | 二羟基二甲基四氢异喹啉-3-甲酰-Lys(Lys-Ala),其合成,活性及应用 | |
CN110577577A (zh) | 1S-甲基-β-四氢咔啉酰-K(GRPAK)其合成,活性和应用 | |
CN107474109B (zh) | 4’-氧乙酰-apak-5-羟基-7-氧乙酰-rgdv-异黄酮,其合成,活性和应用 | |
CN111848730B (zh) | 茶氨酰四氢咪唑并吡啶-6-甲酰极性氨基酸的制备,活性和应用 | |
CN111848728B (zh) | 茶氨酰四氢咪唑并吡啶-6-甲酰-l-组氨酸的制备,活性和应用 | |
CN106432414B (zh) | 五甲氧色胺基-kapkap,其制备,活性和应用 | |
CN106432418B (zh) | 五甲氧色胺基羰丙酰-pakpak,其制备,活性和应用 | |
CN111848606B (zh) | 茶氨酰四氢咪唑并吡啶-6-甲酰甘氨酸和丙氨酸的制备,活性和应用 | |
CN112175041B (zh) | 乙基qrpak修饰的双咔啉并哌嗪二酮,其制备,活性和应用 | |
CN111848724B (zh) | 茶氨酰四氢咪唑并吡啶-6-甲酰酸性氨基酸的制备,活性和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20220809 |