CN106279362A - Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 - Google Patents
Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 Download PDFInfo
- Publication number
- CN106279362A CN106279362A CN201510351775.8A CN201510351775A CN106279362A CN 106279362 A CN106279362 A CN 106279362A CN 201510351775 A CN201510351775 A CN 201510351775A CN 106279362 A CN106279362 A CN 106279362A
- Authority
- CN
- China
- Prior art keywords
- val
- leu
- cys
- arg
- boc
- 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.)
- Granted
Links
- 230000015572 biosynthetic process Effects 0.000 title description 2
- 238000003786 synthesis reaction Methods 0.000 title description 2
- 230000000202 analgesic effect Effects 0.000 claims abstract description 13
- 230000002785 anti-thrombosis Effects 0.000 claims abstract description 10
- 239000003146 anticoagulant agent Substances 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 3
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 3
- 239000003527 fibrinolytic agent Substances 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 9
- 238000006555 catalytic reaction Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims description 6
- MVEAAGBEUOMFRX-UHFFFAOYSA-N ethyl acetate;hydrochloride Chemical compound Cl.CCOC(C)=O MVEAAGBEUOMFRX-UHFFFAOYSA-N 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 238000010511 deprotection reaction Methods 0.000 claims description 2
- 238000007327 hydrogenolysis reaction Methods 0.000 claims description 2
- 238000010561 standard procedure Methods 0.000 claims description 2
- 208000007536 Thrombosis Diseases 0.000 abstract description 22
- 230000000694 effects Effects 0.000 abstract description 12
- 230000000259 anti-tumor effect Effects 0.000 abstract description 7
- 229960000103 thrombolytic agent Drugs 0.000 abstract description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 28
- 241000699670 Mus sp. Species 0.000 description 19
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- 239000004698 Polyethylene Substances 0.000 description 7
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 7
- -1 polyethylene Polymers 0.000 description 7
- 229920000573 polyethylene Polymers 0.000 description 7
- 210000003462 vein Anatomy 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 6
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 229960001138 acetylsalicylic acid Drugs 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 240000003186 Stachytarpheta cayennensis Species 0.000 description 5
- 235000009233 Stachytarpheta cayennensis Nutrition 0.000 description 5
- 229920000669 heparin Polymers 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 102000015636 Oligopeptides Human genes 0.000 description 4
- 108010038807 Oligopeptides Proteins 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000000376 reactant Substances 0.000 description 4
- 239000012047 saturated solution Substances 0.000 description 4
- 239000007832 Na2SO4 Substances 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 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 3
- 229960001008 heparin sodium Drugs 0.000 description 3
- 208000020442 loss of weight Diseases 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 230000035807 sensation Effects 0.000 description 3
- 229910052938 sodium sulfate Inorganic materials 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 230000002537 thrombolytic effect Effects 0.000 description 3
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 3
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 2
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 2
- 238000001467 acupuncture Methods 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 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 2
- 230000033228 biological regulation Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 239000012230 colorless oil Substances 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 229920001971 elastomer Polymers 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 229960005356 urokinase Drugs 0.000 description 2
- UUTKICFRNVKFRG-WDSKDSINSA-N (4R)-3-[oxo-[(2S)-5-oxo-2-pyrrolidinyl]methyl]-4-thiazolidinecarboxylic acid Chemical compound OC(=O)[C@@H]1CSCN1C(=O)[C@H]1NC(=O)CC1 UUTKICFRNVKFRG-WDSKDSINSA-N 0.000 description 1
- NWXMGUDVXFXRIG-WESIUVDSSA-N (4s,4as,5as,6s,12ar)-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O NWXMGUDVXFXRIG-WESIUVDSSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 229930195573 Amycin Natural products 0.000 description 1
- MDXGYYOJGPFFJL-QMMMGPOBSA-N N(alpha)-t-butoxycarbonyl-L-leucine Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)OC(C)(C)C MDXGYYOJGPFFJL-QMMMGPOBSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000001715 carotid artery Anatomy 0.000 description 1
- 210000001168 carotid artery common Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003194 forelimb Anatomy 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 210000004731 jugular vein Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 229910000343 potassium bisulfate Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000004761 scalp Anatomy 0.000 description 1
- 238000002444 silanisation Methods 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Classifications
-
- 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
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了Arg-Leu-Val-Cys-Val,公开了它的制备方法,公开了它的镇痛活性,公开了它的抗肿瘤活性,公开了它的抗血栓活性及公开了它的溶血栓活性。因而本发明公开了它在制备镇痛药物、抗肿瘤药物、抗血栓药物和溶血栓药物中的应用。
Description
技术领域
本发明涉及Arg-Leu-Val-Cys-Val,涉及它的制备方法,涉及它的镇痛活性,涉及它的抗肿瘤活性,涉及它的抗血栓活性及涉及它的溶血栓活性。因而本发明涉及它在制备镇痛药物、抗肿瘤药物、抗血栓药物和溶血栓药物中的应用。本发明属于生物医药领域。
背景技术
发明抗肿瘤、抗血栓、溶血栓、抗凝血和镇痛作用寡肽是发明人长期关注的领域。虽然发明人发明公开过一系列具有这些活性的寡肽,但是集这些活性为一体的寡肽一直没有得到。经过近10年的序列设计和近5年的实验研究,发明人发现Arg-Leu-Val-Cys-Val是集镇痛作用,抗炎作用,抗肿瘤作用抗血栓作用和溶血栓作用为一体的寡肽。
发明内容
本发明的第一个内容是提供Arg-Leu-Val-Cys-Val。
本发明的第二个内容是提供Arg-Leu-Val-Cys-Val的合成方法,该方法包括:
(1)在DCC和HOBt的催化下按照标准方法制备Boc-Leu-Val-OBzl;
(2)Boc-Leu-Val-OBzl在Pd/C催化下氢解转化成Boc-Leu-Val
(3)在DCC和HOBt的催化下制备Boc-Cys(Bzl)-Val-OBzl;
(4)Boc-Cys(Bzl)-Ser-OBzl在浓度为4N的氯化氢-乙酸乙酯溶液中,0℃脱Boc,转化成Cys(Bzl)-Val-OBzl
(5)在DCC和HOBt的催化下制备Boc-Arg(NO2)-Leu-Val-OBzl;
(6)Boc-Arg(NO2)-Leu-Val-OBzl在浓度为2N的NaOH水溶液中,0℃转化成Boc-Arg(NO2)-Leu-Val;
(7)在DCC和HOBt的催化下制备Boc-Arg(NO2)-Leu-Val-Cys(Bzl)-Val-OBzl;
(8)Boc-Arg(NO2)-Leu-Val-Cys(Bzl)-Val-OBzl在TFA和TFSMA中,0℃脱保护转化成Arg-Leu-Val-Cys-Val。
本发明的第三个内容是评价Arg-Leu-Val-Cys-Val的镇痛作用。
本发明的第四个内容是评价Arg-Leu-Val-Cys-Val的抗肿瘤作用。
本发明的第五个内容是评价Arg-Leu-Val-Cys-Val的抗血栓作用。
本发明的第六个内容是评价Arg-Leu-Val-Cys-Val的溶血栓作用。
附图说明
图 1.Arg-Leu-Val-Cys-Val的合成路线.i)二环己基碳二亚胺(DCC),1-羟基苯并三唑(HOBt),N-甲基吗啉(NMM),四氢呋喃(THF);ii)浓度为4N的氯化氢-乙酸乙酯溶液,0℃;iii)H2,Pd/C;iv)TFA,TFMSA,0℃。
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Boc-Leu-Val-OBzl
冰浴下2.31g(10mmol)Boc-Leu溶解于10ml无水THF,加入1.36g(10mmol)HOBt,加入2.47g(12mmol)DCC与10ml无水THF的溶液,活化30分钟,加入2.07g(10mmol)Val-OBzl,用NMM调节pH=9,反应结束后过滤除去二环己基脲(DCU)。滤液减压浓缩,残留物用乙酸乙酯溶解,再次过滤DCU,滤液用NaHCO3饱和溶液洗3遍,用NaCl饱和溶液洗3遍,5%的KHSO4溶液洗3遍,NaCl饱和溶液洗3遍,5%的NaHCO3溶液萃洗3遍,NaCl饱和溶液萃洗3遍,乙酸乙酯层用无水Na2SO4干燥12小时,滤除Na2SO4,滤液减压浓缩,得到5.2g(100%)标题化合物,为黄色油状物。ESI-MS(m/e):421[M+H]+。
实施例2制备Leu-Val-OBzl
冰浴下将4.20g(10mmol)Boc-Leu-Val-OBzl用10ml干燥过的乙酸乙酯溶解,搅拌10min,加入50mL氯化氢-乙酸乙酯溶液(4N)反应4h,原料点消失。反应混合物减压浓缩至干,残留物加40mL干燥过的乙酸乙酯溶解,得到的溶液减压浓缩至干。残留物按该操作重复3次。残留物加无水乙醚,用塑料铲研磨,减压浓缩除去乙醚。残留物按该操作重复3次。得到3.02g(97%)标题化合物,为无色粉末。ESI+-MS(m/e):322[M+H]+。
实施例3制备Boc-Arg(NO2)-Leu-Val-OBzl
按照实施例1的方法从3.19g(10mmol)Boc-Arg(NO2)和3.21g(10mmol)Leu-Val-OBzl得到3.02g(49%)标题化合物,为无色粉末。ESI+-MS(m/e):623 [M+H]+。
实施例4制备Boc-Arg(NO2)-Leu-Val
冰浴下将6.21g(10mmol)Boc-Arg(NO2)-Leu-Val-OBzl溶解于甲醇,搅拌10min,逐滴加入2N NaOH溶液,调节pH到13-14。保持冰浴反应4个小时,反应完全。冰浴下反应液用饱和KHSO4溶液调节pH到中性,先减压浓缩除甲醇,再用饱和KHSO4溶液调节pH到2。反应液用乙酸乙酯萃取3遍。得到的溶液用饱和NaCl溶液洗至中性,用无水Na2SO4干燥12小时,过滤,滤液减压浓缩,得到4.93g(100%)标题化合物,为无色油状物。ESI-MS(m/e):532[M+H]+。
实施例5制备Boc-Cys(Bzl)-Val-OBzl
按照实施例1的方法从3.11g(10mmol)Boc-Cys(Bzl)和2.07g(10mmol)Val-OBzl得到4.23g(95%)标题化合物,为无色油状物。ESI+-MS(m/e):539[M+K。
实施例6制备Cys(Bzl)-Val-OBzl的制备
按照实施例2的方法从5.0g(10mmol)Boc-Cys(Bzl)-Val-OBzl得到3.9g(98%)标题化合物,为无色粉末。ESI+-MS(m/e):401[M+H]+。
实施例7制备Boc-Arg(NO2)-Leu-Val-Cys(Bzl)-Val-Obzl
按照实施例1的方法从5.37g(10mmol)Boc-Arg(NO2)-Leu-Val和4.0g(10mmol)Cys(Bzl)-Val-OBzl得到3.67g(40%)标题化合物,为无色果冻状物。ESI+-MS(m/e):915[M+H]+;1H-NMR(300MHz,DMSO-d6):δ/ppm=8.32(d,1H),8.15(d,1H),7.83(m,2H),7.32(m,10H),7.22(m,1H),6.95(d,1H),5.12(s,2H),4.67(t,1H),4.40(t,2H),4.22(m,2H),3.90(m,1H),3.75(s,2H),3.11(s,2H),2.72(m,1H),2.53(m,2H),1.98(m,2H),1.70-1.40(m,6H),1.37(s,9H),0.87(s,3H),0.85(s,3H),0.84(s,3H),0.83(s,3H),0.80(s,3H),0.78(s,3H)。
实施例8制备Arg-Leu-Val-Cys-Val
冰浴下将456mg(0.5mmol)Boc-Arg(NO2)-Leu-Val-Cys(Bzl)-Val-OBzl用12mL三氟醋酸溶解,搅拌10min,加入3mL TFMSA(三氟甲磺酸)反应30min后,原料点消失。反应混合物用无水乙醚稀释,用塑料铲磨洗,倾倒乙醚。残留物再用无水乙醚稀释,用塑料铲磨洗,倾倒乙醚。该操作重复3次。得到的棕黄色粉末用三蒸水溶解,用氨水调pH至7,用Sephdex G10柱纯化,收集的馏分冻干,得到173mg(62%)标题化合物。ESI-MS(m/e):590[M+H]+;Mp 174.2~174.9℃.(c=0.08,甲醇).IR:3326,2964,2257,1786,1650,1523,1407,1364,1275,1245,1224,1160,1027,1004,863,824,760,710,637.1H-NMR(300MHz,DMSO-d6):δ/ppm=8.46(m,1H), 8.06(m,4H),7.38(m,5H),4.49(m,1H),4.19(m,2H),3.77(m,3H),3.00(m,4H),2.10(m,2H),1.63(m,2H),1.45(m,3H),1.21(m,3H),0.85(m,18H)。
实施例9评价Arg-Leu-Val-Cys-Val的镇痛活性
雄性ICR小鼠(20±2g)装到小鼠固定器中,鼠尾暴露于固定器外,于鼠尾距离尾尖三分之一处标记,作为光感感受器的感应点,照射鼠尾距离尾尖三分之二的位置。测痛仪预热30min,记时,将小鼠鼠尾遮住光感仪。灯闪烁时开始记时,鼠尾离开光感仪时停止记时,测得的时间即为小鼠产生痛感的时间,连续测定3次,每次测量间隔为5min,取平均值。未服药小鼠产生痛感的时间为基础痛感时间。服药引起的小鼠产生痛感时间变化反映了药物的镇痛活性。小鼠,随机分组,每组14只,测定基础痛感时间后,小鼠或口服0.2mL Arg-Leu-Val-Cys-Val(RLVCV)的生理盐水溶液,剂量为1μmol/kg或口服0.2mL生理盐水或口服阿司匹林的生理盐水溶液,剂量为1200μmol/kg。测量给药后30,60,90,120,150和180min小鼠产生痛感的时间。数据用均值±SD秒表示,列如表 1,用方差分析并进行t检验。数据表明,口服生理盐水30,60,90,120,150和180min之后小鼠产生痛感的时间与口服生理盐水之前的基础痛感时间没有显著差异;口服1200μmol/kg阿司匹林60,90,120,150和180min之后小鼠产生痛感的时间显著长于口服阿司匹林之前的基础痛感时间;口服1μmol/kgArg-Leu-Val-Cys-Val 30,60,90,120,150和180min之后小鼠产生痛感的时间显著长于口服Arg-Leu-Val-Cys-Val之前的基础痛感时间。可见,口服1μmol/kgArg-Leu-Val-Cys-Val60,90,120,150和180min之后的镇痛活性与口服1200μmol/kg阿司匹林相当。而口服1μmol/kgArg-Leu-Val-Cys-Val30min之后的镇痛活性比口服1200μmol/kg阿司匹林还强。
表 1Arg-Leu-Val-Cys-Val(RLVCV)的镇痛活性
n=14;a)与基础痛感时间比p>0.05;b)与基础痛感时间比p<0.05;c)与基础痛感时间比p<0.05.
实施例10评价Arg-Leu-Val-Cys-Val的抗肿瘤活性
无菌条件下取接种于ICR小鼠7-10天的S180肉瘤,加入适量生理盐水配制成瘤细胞悬液,细胞数为2×107/mL,接种于健康雄性ICR小鼠前肢腋皮下,每只小鼠注射0.2mL。肿瘤接种24h后,小鼠每日腹腔注射0.2mLArg-Leu-Val-Cys-Val的生理盐水溶液,连续给药10天,剂量为1μmol/kg;或小鼠每日腹腔注射0.2mL阿霉素的生理盐水溶液,连 续给药10天,剂量为2μmol/kg;或小鼠每日腹腔注射0.2mL生理盐水,连续给药10天。实验进行至第11天,称小鼠体重,乙醚麻醉剖取各组小鼠的肿瘤,称重并以瘤重表示化合物的活性,数据列入表 3。结果表明1μmol/kg Arg-Leu-Val-Cys-Val治疗小鼠的瘤重明显小于生理盐水治疗小鼠的瘤重,Arg-Leu-Val-Cys-Val显示确切的抗肿瘤活性。
表 2Arg-Leu-Val-Cys-Val对S180荷瘤小鼠的瘤重的影响
n=15;瘤重为均值±SD g;a)与生理盐水比p<0.01。
实施例11评价Arg-Leu-Val-Cys-Val的抗血栓活性
1)将聚乙烯管拉成一端为斜口的细管,定长为10.0cm,分别为右经静脉(管径较粗)及左颈动脉(管径较细)插管;中段聚乙烯管定长为8.0cm,血栓线压在颈动脉插管方向,插管前需在管中充满肝素。
2)体重250±雄性大鼠口服1μmol/kgArg-Leu-Val-Cys-Val或167μmol/kg阿司匹林或0.3mL/100g生理盐水30分钟后,腹腔注射20%的乌拉坦进行麻醉。仰卧位将大鼠固定于鼠板上,剪开颈部皮肤,分离右颈总动脉及左颈静脉,血管下压线,结扎远心端,静脉靠远心端处剪一小口,进行静脉端插管,注射肝素,系线固定,再用动脉夹夹住动脉近心端,靠近远心端方向剪一小口,进行动脉端结扎,系线固定后松开动脉夹,建立体外循环旁路。循环15分钟后先剪断静脉端观察血液循环是否正常,若正常从动脉端取出血栓线,在纸上沾干浮血后称重,以栓重表示化合物的活性,数据列入表 4。结果表明,Arg-Leu-Val-Cys-Val治疗大鼠的血栓重明显小于生理盐水治疗大鼠的血栓重,Arg-Leu-Val-Cys-Val显示确切的抗血栓活性。。
表 3Arg-Leu-Val-Cys-Val的抗栓活性
n=12;a)与生理盐水比p<0.05.
实施例12评价Arg-Leu-Val-Cys-Val的溶栓活性
将大鼠静息饲养一天,随机分组,每组12只,禁食不禁水。
1)将体重250±雄性SD大鼠用20%乌拉坦溶液进行麻醉(6mL/kg腹腔)。麻醉后的大鼠固定于鼠板上,分离右侧颈总动脉,于近心端夹动脉夹,近心端和远心端分别传入手术线,在远心端插取血管,松开动脉夹取约1mL动脉血。迅速将动脉血注射入垂直的造栓管(长16mm,内径2.5mm,外径5mm,管底用1mL EP管底座)中,(注意不可有气泡)每个造栓管中注入0.1mL大鼠动脉血液,往管内迅速插入血栓固定螺栓(长20mm,),血液凝固40min,用针灸针取出血栓,称取血栓重量。
2)旁路插管由三部分组成,其中中段为聚乙烯胶管,长60mm,内径3.5mm,两端为相同的聚乙烯管,长100mm,内径1mm,外景2mm,该管的一段拉成尖管,另一端的外部套一段长7mm,外径3.5mm的聚乙烯管。三段管的内壁均为硅烷化。
3)将血栓包裹的血栓固定螺栓置于中段的聚乙烯胶管内,胶管的两端分别与两根聚乙烯的加粗端相套,用注射器通过尖管端注满肝素生理盐水溶液(50IU/kg)。将充满肝素钠的生理盐水溶液一端插入左侧静脉,另一端用注射器加入准确量的肝素钠抗凝后,拔掉肝素钠的注射器,插入动脉端。用头皮针将生理盐水(3mL/kg)或者尿激酶的生理盐水溶液(20000IU/kg)或Arg-Leu-Val-Cys-Val的生理盐水溶液(1μmol/kg)的通过旁路管的中段,刺入远离血栓固定螺栓的近静脉处,打开动脉夹,使血流通过旁路管道从动脉流入静脉的时刻为循环起始时间,缓慢的将注射器中的液体注入到血液中(约6min),使生理盐水,尿激酶,Arg-Leu-Val-Cys-Val通过血液循环,按静脉-心脏-动脉的顺序作用到血栓上。60min后取出附有血栓的螺栓,蘸去浮血,记录重量。以血栓减重代表溶栓活性。数据列入表 4。结果表明,Arg-Leu-Val-Cys-Val治疗大鼠的血栓减重明显大于生理盐水治疗大鼠的血栓减重,Arg-Leu-Val-Cys-Val显示确切的溶血栓活性。
表 4Arg-Leu-Val-Cys-Val的溶栓活性
n=10;a)与生理盐水比p<0.01。
Claims (6)
1.Arg-Leu-Val-Cys-Val。
2.权利要求1的Arg-Leu-Val-Cys-Val的制备方法,该方法包括:
(1)在DCC和HOBt的催化下按照标准方法制备Boc-Leu-Val-OBzl;
(2)Boc-Leu-Val-OBzl在Pd/C催化下氢解转化成Boc-Leu-Val;
(3)在DCC和HOBt的催化下制备Boc-Bzl保护巯基的Cys-Val-OBzl;
(4)Boc-Bzl保护巯基的Cys-Ser-OBzl在浓度为4N的氯化氢-乙酸乙酯溶液中,0℃脱Boc转化成Bzl保护巯基的Cys-Val-OBzl;
(5)在DCC和HOBt的催化下制备Boc-NG-NO2-Arg-Leu-Val-OBzl;
(6)Boc-NG-NO2-Arg-Leu-Val-OBzl在浓度为2N的NaOH水溶液中0℃水解为Boc-NG-NO2-Arg-Leu-Val;
(7)在DCC和HOBt的催化下制备Boc-NG-NO2-Arg-Leu-Val-Bzl保护巯基的Cys-Val-OBzl;
(8)Boc-NG-NO2-Arg-Leu-Val-Bzl保护巯基的Cys-Val-OBzl在TFA和TFSMA中,0℃脱保护转化成Arg-Leu-Val-Cys-Val。
3.权利要求1的Arg-Leu-Val-Cys-Val在制备抗肿瘤药物中的应用。
4.权利要求1的Arg-Leu-Val-Cys-Val在制备镇痛药物中的应用。
5.权利要求1的Arg-Leu-Val-Cys-Val在制备抗血栓药物中的应用。
6.权利要求1的Arg-Leu-Val-Cys-Val在制备溶栓药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510351775.8A CN106279362B (zh) | 2015-06-23 | 2015-06-23 | Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510351775.8A CN106279362B (zh) | 2015-06-23 | 2015-06-23 | Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN106279362A true CN106279362A (zh) | 2017-01-04 |
CN106279362B CN106279362B (zh) | 2019-07-12 |
Family
ID=57650894
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510351775.8A Expired - Fee Related CN106279362B (zh) | 2015-06-23 | 2015-06-23 | Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106279362B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993022340A1 (en) * | 1992-04-30 | 1993-11-11 | Schering Corporation | Modulation of thrombospondin-cd36 interactions |
CN1099040A (zh) * | 1993-04-01 | 1995-02-22 | 默克专利股份有限公司 | 线性粘合抑制剂 |
CN101434649A (zh) * | 2008-12-12 | 2009-05-20 | 余榕捷 | 一种短肽及其制备方法和在诊断或治疗与pacap、vip相关疾病中的应用 |
WO2014120837A2 (en) * | 2013-01-29 | 2014-08-07 | The Regents Of The University Of California | Pretargeted activatable cell penetrating peptide with intracellulary releaseable prodrug |
-
2015
- 2015-06-23 CN CN201510351775.8A patent/CN106279362B/zh not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1993022340A1 (en) * | 1992-04-30 | 1993-11-11 | Schering Corporation | Modulation of thrombospondin-cd36 interactions |
CN1099040A (zh) * | 1993-04-01 | 1995-02-22 | 默克专利股份有限公司 | 线性粘合抑制剂 |
CN101434649A (zh) * | 2008-12-12 | 2009-05-20 | 余榕捷 | 一种短肽及其制备方法和在诊断或治疗与pacap、vip相关疾病中的应用 |
WO2014120837A2 (en) * | 2013-01-29 | 2014-08-07 | The Regents Of The University Of California | Pretargeted activatable cell penetrating peptide with intracellulary releaseable prodrug |
Also Published As
Publication number | Publication date |
---|---|
CN106279362B (zh) | 2019-07-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105218629A (zh) | 异喹啉-3-甲酰-RC-OBzl,其制备,纳米结构,活性和应用 | |
CN105198960B (zh) | 咪唑并吡啶-6-甲酰-Met-AA-OBzl,其合成,活性和应用 | |
CN106317185B (zh) | His-Gly-Tyr-Asp,其合成,活性和应用 | |
CN106279362A (zh) | Arg-Leu-Val-Cys-Val,其合成,药理活性和应用 | |
CN106317191A (zh) | Thr-Val-Gly-Cys-Ser,其合成,活性和应用 | |
CN108929372A (zh) | 1R-甲基-β-四氢咔啉酰-K(GRPAK)-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 | |
CN106317193B (zh) | 含Tyr-Val片段的肽,其合成、活性和应用 | |
CN108948146A (zh) | 1R-甲基-β-四氢咔啉酰-K(ARPAK)-RGDV, 其合成, 活性和应用 | |
CN105315334A (zh) | 异喹啉-3-甲酰-RF-OBzl,其制备,纳米结构,活性和应用 | |
CN105175492A (zh) | 咪唑并吡啶-6-甲酰-Met-Pro-OBzl,其合成,活性和应用 | |
CN107488213B (zh) | 华法林-4-o-乙酰-yigsk,其合成,药理活性和应用 | |
CN106317177A (zh) | Gly-Phe-Pro,其合成,活性和应用 | |
CN106317172A (zh) | Pro-Ser,其合成,活性和应用 | |
CN105315325A (zh) | 咪唑并吡啶-6-甲酰-Met-Arg(NO2)-OBzl,其合成,活性和应用 | |
CN108948145A (zh) | 1R-甲基-β-四氢咔啉酰-K(PAK)-RGDV,其合成,活性和应用 | |
CN108948155A (zh) | 1R-甲基-β-四氢咔啉酰-K(QRPAK)-RGDV, 其合成, 活性和应用 | |
CN107488210B (zh) | 华法林-4-o-乙酰-二肽,其合成,活性和应用 | |
CN107459557A (zh) | 左旋维c-2-氧乙酰-grpak,其合成,活性和应用 | |
CN106317187B (zh) | 五甲氧色胺基羰丙酰-pak肽,其制备,活性和应用 | |
CN106336450B (zh) | RGDV-The-GRPAK十肽,其制备,活性及应用 | |
CN107488214B (zh) | 华法林-4-o-乙酰-lpniskp其合成,活性和应用 | |
CN106349333B (zh) | The-K(KAPRG)RGDV十一肽,其制备,活性及应用 | |
CN106317198B (zh) | GRPAK-The-RGDV,其制备,活性及应用 | |
CN106317189B (zh) | 五甲氧色胺基羰丙酰-rpak肽,其制备,活性和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into 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: 20190712 |