Pro-Ala-Lys-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3、及其合成和作为溶血栓剂的应用转让专利
申请号 : CN201010573521.8
文献号 : CN102477069B
文献日 : 2013-07-24
发明人 : 赵明 , 彭师奇 , 金绍明
申请人 : 首都医科大学
摘要 :
权利要求 :
1.具有溶血栓活性的寡肽,其结构式为通式I所示: Pro-Ala-Lys-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3; 其中,n=6、8、10、12、14或16。
2.一种合成权利要求1所述寡肽的方法,包括以下步骤:
1)Boc-Pro与Ala-OBzl缩合为Boc-Pro-Ala-OBzl;
2)将Boc-Pro-Ala-OBzl氢解为Boc-Pro-Ala;
3)Boc-Pro-Ala与Lys(Boc)-OBzl缩合为Boc-Pro-Ala-Lys(Boc)-OBzl;
4)将Boc-Pro-Ala-Lys(Boc)-OBzl氢解为Boc-Pro-Ala-Lys(Boc);
5)Boc-Asp与饱和脂肪醇缩合为
Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3,其中n=6、8、10、12、14或16;
6)Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3脱去Boc生成 Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3;
7)Boc-Pro-Ala-Lys(Boc)与Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3缩合为Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3;
8)Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)n-CH3]-OCH2(CH2)nCH3脱除Boc,即得; 其中,步骤(1)中在DCC和HOBt存在下,Boc-Pro在无水THF中与Ala-OBzl缩合为Boc-Pro-Ala-OBzl; 步骤2)中在甲醇中将Boc-Pro-Ala-OBzl氢解为Boc-Pro-Ala; 步骤3)中在DCC和HOBt存在下Boc-Pro-Ala在无水THF中与Lys(Boc)-OBzl缩合为Boc-Pro-Ala-Lys(Boc)-OBzl; 步骤4)中在甲醇中将Boc-Pro-Ala-Lys(Boc)-OBzl氢解为 Boc-Pro-Ala-Lys(Boc);
步骤5)中在DCC存在下Boc-Asp在无水THF中与饱和脂肪醇缩合为 Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3; 步骤6)中在氯化氢-乙酸乙酯溶液中
Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3脱去Boc生成 Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3; 步骤7)中在DCC和HOBt存在下Boc-Pro-Ala-Lys(Boc)在无水THF中与 Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3缩合为Boc-Pro-Ala-Lys(Boc)- Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3; 步骤8)中在氯化氢-乙酸乙酯溶液中
Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)n-CH3]-OCH2(CH2)nCH3脱除Boc。
3.权利要求1所述的寡肽在制备溶血栓药物中的用途。
说明书 :
Pro-Ala-Lys-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3、及其合
成和作为溶血栓剂的应用
技术领域
背景技术
发明内容
Boc-Pro-Ala-OBzl氢解为Boc-Pro-Ala;步骤3)中优选在DCC和HOBt存在下Boc-Pro-Ala在无水THF中与Lys(Boc)-OBzl缩合为Boc-Pro-Ala-Lys(Boc)-OBzl;步骤4)中优选在甲醇中将Boc-Pro-Ala-Lys(Boc)-OBzl氢解为Boc-Pro-Ala-Lys(Boc);步骤5)中优选在DCC存在下Boc-Asp在无水THF中与饱和脂肪醇缩合为Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3;步骤6)中优选在氯化氢-乙酸乙酯溶液中Boc-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3脱去Boc生成Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3;步骤7)中优选在DCC和HOBt存在下Boc-Pro-Ala-Lys(Boc)在无水THF中与Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3缩合为Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)nCH3]-OCH2(CH2)nCH3;步骤8)中优选在氯化氢-乙酸乙酯溶液中Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)n-CH3]-OCH2(CH2)nCH3脱除Boc。
附图说明
具体实施方式
反应混合物过滤,滤除二环己基脲(DCU)。滤液减压浓缩,除去THF。残留物用250ml乙酸乙酯溶解。得到的溶液依次用饱和NaHCO3水溶液洗、饱和NaCl水溶液洗、5%KHSO4水溶液洗和饱和NaCl水溶液洗。乙酸乙酯相用无水Na2SO4干燥、过滤、滤液减压浓缩至干,残留物+
经柱层析纯化,得到4.23g(72%)目标化合物,为无色粉末。ESI-MS(m/e):458[M+H].[0034] 实施例2制备Boc-Asp[OCH2(CH2)8CH3]-[OCH2(CH2)8CH3](1b)
+
ESI-MS(m/e):358[M+H].
制得3.59g(91%)目标化合物,为黄色粉末。ESI-MS(m/e):285[M-H].
[α]D = -701(c = 0.44,CH3OH);ESI-MS(m/e):854[M+H] ;IR(KBr):3285,3250,3242,
3208,3071,2955,2922,2859,1740,1717,1692,1636,1526,1479,1454,1408,1366,1321,
1
1273,1252,1221,1165,1136,1128,1096,1070,1011,777.H NMR(300MHz,CDCl3):δ/ppm=
7.44-7.32(m,1H),7.24-7.11(m,2H),7.02(s,1H),4.94(s,1H),4.81-4.77(m,1H),4.39(d,J=4.8Hz,2H),4.22(s,1H),4.08-3.97(m,4H),3.45-3.41(m,2H),3.04(d,J=5.1Hz,2H),
2.98-2.75(m,2H),2.08(m,2H),1.97-1.86(m,3H),1.67-1.58(m,5H),1.42-1.14(m,44H),
13
0.85(t,J= 5.1Hz,J =6.9Hz,6H). C NMR(75MHz,CDCl3):δ/ppm = 172.64,172.16,
171.67,171.28,170.72,170.44,170.28,156.12,155.72,141.21,128.36,127.31,126.88,
126.64,125.48,80.68,78.86,65.76,65.29,65.18,64.94,60.46,53.01,49.46,48.66,
47.20,40.09,36.22,31.71,31.49,29.54,29.37,29.15,29.10,28.45,28.40,28.28,
25.80,25.71,24.54,22.56,17.59,15.19,14.00.
25 +
色粉末。Mp 94℃;[α]D =-541(c=0.58,CH3OH);ESI-MS(m/e):910[M+H] ;IR(KBr):
3289,3277,3237,3063,3053,2953,2928,2857,1740,1684,1636,1526,1456,1402,1393,
1
1366,1275,1248,1217,1165,1126,1099,988,914,864,692.H NMR(300MHz,CDCl3):δ/ppm=7.35(s,1H),7.14-6.99(m,3H),4.93(s,1H),4.82-4.81(m,1H),4.41(d,J=4.5Hz,
2H),4.24(s,1H),4.09-4.02(m,4H),3.44(s,2H),3.06(s,2H),2.95(d,J = 15.9Hz,
1H),2.81(d,J = 9.9Hz,1H),2.10(m,2H),1.88(s,3H),1.59(s,6H),1.44-1.35(m,24H),
13
1.25(s,29H),0.86-0.84(m,6H). C NMR(75MHz,CDCl3):δ/ppm=172.59,172.06,171.59,
171.22,170.78,170.46,156.08,128.42,127.39,126.91,80.78,78.85,77.30,65.91,
65.35,65.22,65.05,60.45,52.99,49.46,48.64,47.26,40.07,36.24,31.84,31.51,
29.50,29.40,29.27,29.24,29.19,28.90,28.48,28.42,28.30,25.83,25.74,24.57,
23.07,22.63,22.27,17.53,14.06.
25 +
无色粉末。Mp 91℃;[α]D =-439(c=0.72,CH3OH);ESI-MS(m/e):966[M+H] ;IR(KBr):
3970,3296,3277,3244,2957,2924,2855,1728,1684,1636,1530,1506,1456,1398,1395,
1
1366,1275,1252,1206,1177,1119.H NMR(300MHz,CDCl3):δ/ppm = 7.25-7.10(m,
2H),7.00-6.86(m,1H),4.94(s,1H),4.82-4.77(m,1H),4.41-4.39(m,2H),4.23(s,1H),
4.09-4.00(m,4H),3.43(s,2H),3.04(d,J = 4.8Hz,2H),2.95-2.90(m,1H),2.81-2.77(m,
1H),2.08-2.06(m,2H),1.86(s,3H),1.67-1.57(m,5H),1.43-1.40(m,18H),1.34(d,J =
13
6.9Hz,4H),1.23(s,35H),0.85(t,J=5.1Hz,J=6.9Hz,6H). C NMR(75MHz,CDCl3):δ/ppm = 172.56,172.10,171.65,171.24,170.81,170.73,170.44,170.12,156.06,155.71,
141.25,128.36,127.31,126.88,80.67,78.77,65.97,65.88,65.29,65.18,64.95,60.44,
52.98,52.87,49.38,48.63,47.21,40.06,36.23,31.85,31.49,29.60,29.57,29.55,
29.48,29.39,29.29,29.23,29.18,28.91,28.47,28.41,28.28,25.82,25.73,24.54,
23.01,22.62,22.28,17.58,14.04.
25 +
为无色粉末。Mp 80℃;[α]D =-626(c=0.50,CH3OH);ESI-MS(m/e):1022[M+H] ;
1
IR(KBr):2955,2926,2855,1728,1684,1638,1404,1395,1366,1304,1169.H NMR(300MHz,CDCl3):δ/ppm = 7.05-6.99(m,2H),4.89-4.79(m,2H),4.47-4.40(m,2H),4.25(s,1H),
4.13-4.05(m,4H),3.45(s,2H),3.08-3.06(m,2H),2.98-2.93(m,1H),2.85-2.75(m,1H),
2.28-2.08(m,2H),1.86(s,3H),1.72-1.60(m,5H),1.45-1.36(m,22H),1.28-1.17(m,44H),
13
0.85(t,J= 5.7Hz,J =6.9Hz,6H). C NMR(75MHz,CDCl3):δ/ppm = 172.57,172.04,
171.54,171.19,170.86,170.78,170.45,170.40,170.03,156.13,156.07,80.77,78.85,
66.04,65.92,65.35,60.49,53.00,52.89,49.41,48.65,48.51,47.23,40.13,36.25,
36.14,32.29,31.89,31.49,29.65,29.63,29.58,29.51,29.41,29.32,29.25,29.21,
28.83,28.50,28.43,28.30,25.85,25.76,24.56,23.09,22.65,22.26,17.54,14.07.[0072] 实 施 例21制 备 Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)14CH3]-[OCH2(CH2)14CH3](3e)
25 +
为无色粉末。Mp 74℃;[α]D =-907(c=0.34,CH3OH);ESI-MS(m/e):1078[M+H] ;
IR(KBr):3304,3289,2955,2920,2851,1738,1688,1639,1537,1468,1456,1398,1393,
1
1366,1339,1292,1279,1229,1171,1119.H NMR(300MHz,CDCl3):δ/ppm=7.09-7.02(m,
3H),4.92-4.84(m,2H),4.42(s,2H),4.26(s,1H),4.11-4.03(m,4H),3.46(s,2H),3.08(s,
2H),2.97(d,J = 16.2Hz,1H),2.83(d,J = 14.4Hz,1H),2.12-1.89(m,5H),1.61(s,6H),
13
1.45-1.37(m,25H),1.26(s,53H),0.87-0.86(m,6H). C NMR(75MHz,CDCl3):δ/ppm =
172.68,172.15,171.57,171.26,170.88,170.80,170.45,170.39,170.27,156.16,155.86,
141.40,128.42,126.68,125.67,117.89,110.75,80.81,78.92,65.95,65.38,65.25,
60.51,53.06,49.50,48.68,48.55,47.28,40.10,36.25,31.90,31.45,29.68,29.64,
29.59,29.52,29.38,29.33,29.27,29.22,28.93,28.50,28.43,28.31,25.85,25.76,
24.59,22.66,17.51,14.08.
25 +
无色粉末。Mp 88℃;[α]D =-526(c=0.60,CH3OH);ESI-MS(m/e):1134[M+H] ;IR(KBr):
3306,3289,3233,2957,2928,2918,2853,1740,1690,1667,1636,1541,1533,1468,1456,
1
1395,1366,1339,1277,1250,1227,1169,1138,1123,1076,772,721.H NMR(300MHz,CDCl3):δ/ppm = 7.06-6.98(m,3H),4.81(m,1H),4.41(m,2H),4.25(s,1H),4.10-4.02(m,
4H),3.45(s,2H),3.06(m,2H),2.93(m,1H),2.83(m,1H),2.16-2.10(m,2H),1.99-1.88(m,
4H),1.60(m,6H),1.45-1.42(m,20H),1.36(d,J = 6.9Hz,4H),1.24(s,60H),0.86(t,J =
13
5.1Hz,J=6.6Hz,6H). C NMR(75MHz,CDCl3):δ/ppm=172.56,172.06,171.58,171.20,
170.84,170.76,170.45,170.06,156.07,155.79,80.73,78.81,65.90,65.34,65.21,
60.47,53.00,49.39,48.68,48.65,48.52,47.25,40.09,38.94,38.71,36.25,34.09,
33.76,31.89,29.67,29.63,29.59,29.52,29.41,29.32,29.26,29.21,28.50,28.43,
28.30,25.85,25.76,22.65,14.07.
D = -725(c = 0.44,CHCl3);ESI-MS(m/e):654[M+H] ;IR(KBr):3456,3426,3360,
3237,3208,3046,2957,2928,2859,2735,1740,1655,1539,1460,1389,1368,1285,1213,
1
1188,1053.H NMR(300MHz,DMSO-d6):δ/ppm = 10.23(s,1H),8.84(d,J = 6.9Hz,1H),
8.48-8.41(m,2H),8.12-8.09(m,3H),4.66-4.59(m,1H),4.40-4.33(m,1H),4.26-4.22(m,
2H),4.02-3.95(m,4H),3.02(s,2H),2.81-2.66(m,4H),2.35-2.23(m,1H),1.91-1.87(m,
13
3H),1.66-1.53(m,8H),1.25(s,25H),0.86(t,J=4.5Hz,J=6.9Hz,6H). C NMR(75MHz,DMSO-d6):δ/ppm = 171.93,171.82,170.90,170.31,168.26,65.27,64.80,59.03,52.53,
49.10,48.91,46.02,38.85,36.28,32.61,31.74,31.67,30.12,29.04,28.48,26.85,
25.78,25.68,23.99,22.52,22.46,22.28,18.30,14.37.C,62.94%;H,9.81%;N,10.49%;
O,16.77%.
D = -554(c = 0.58,CHCl3);ESI-MS(m/e):710[M+H] ;IR(KBr):3372,3227,3026,
1
2955,2926,2907,2855,1744,1647,1533,1395,1173.H NMR(300MHz,DMSO-d6):δ/ppm= 10.22(s,1H),8.84(d,J = 5.7Hz,1H),8.49(s,1H),8.41(d,J = 7.8Hz,1H),8.11(d,J = 6.9Hz,4H),4.66-4.59(m,1H),4.36(t,J = 6.9Hz,J = 13.8Hz,1H),4.26-4.22(m,
2H),4.08-4.01(m,4H),3.21(s,2H),2.73(s,4H),2.32-2.23(m,1H),1.91-1.87(m,4H),
13
1.57-1.53(m,7H),1.26(s,33H),0.85(t,J = 4.5Hz,J = 6.9Hz,6H). CNMR(75MHz,DMSO-d6):δ/ppm = 171.93,171.82,170.88,170.30,168.26,65.27,64.80,59.03,52.53,
49.10,48.90,46.03,38.90,36.30,31.75,30.12,29.41,28.49,26.86,25.78,25.70,
23.99,22.55,22.46,21.52,18.31,14.37.C,64.70%;H,10.16%;N,9.67%;O,15.47%.[0080] 实施例25制备Pro-Ala-Lys-Asp[OCH2(CH2)10CH3]-[OCH2(CH2)10CH3](4c)[0081] 按照实施例7的方法由345mg(0.36mmol)Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2)10CH3]-[OCH2(CH2)10CH3]制得270mg(90%)目标化合物,为无色粉末。Mp 121℃;
25 +
[α]D = -394(c = 0.72,CHCl3);ESI-MS(m/e):766[M+H] ;IR(KBr):3401,3296,3223,
1
3044,2955,2924,2853,2731,1740,1647,1547,1533,1464,1395,1288,1217,1173. H NMR(300MHz,DMSO-d6):δ/ppm = 8.80(d,J = 7.2Hz,1H),8.41(d,J = 7.8Hz,1H),
8.14-8.04(m,3H),4.69-4.59(m,1H),4.38-4.31(m,1H),4.26-4.19(m,2H),4.02-4.00(m,
4H),3.20-3.18(m,2H),2.74-2.72(m,4H),2.30-2.28(m,1H),1.86(s,3H),1.54(s,9H),
13
1.25(s,41H),0.86(t,J = 5.4Hz,J = 6.6Hz,6H). C NMR(75MHz,DMSO-d6):δ/ppm =
171.93,171.80,170.88,170.30,168.27,65.28,64.81,59.07,52.51,49.06,48.89,46.07,
38.88,36.32,31.76,30.09,29.47,29.41,29.17,29.13,28.49,26.90,25.78,25.71,
23.98,22.54,22.46,18.30,14.37.C,66.20%;H,10.47%;N,8.98%;O,14.36%.[0082] 实施例26制备Pro-Ala-Lys-Asp[OCH2(CH2)12CH3]-[OCH2(CH2)12CH3](4d)[0083] 按照实施例7的方法由185mg(0.18mmol)Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH
2)12CH3]-[OCH2(CH2)12CH3]制得152mg(94%)目标化合物,为无色粉末。Mp 109℃;[α]
25 +
D =-580(c = 0.50,CHCl3);ESI-MS(m/e):822[M+H] ;IR(KBr):3944,3433,3298,3229,
1
2922,2855,2749,1742,1647,1545,1462,1387,1344,1292,1221,1169,1067,1003,723.H NMR(300MHz,DMSO-d6):δ/ppm = 10.07(s,1H),8.39(d,J = 7.8Hz,1H),8.12(d,J =
7.8Hz,1H),4.66-4.62(m,1H),4.36-4.31(m,1H),4.26-4.18(m,2H),4.02-3.99(m,4H),
3.24-3.17(m,2H),2.74-2.72(m,4H),2.34-2.29(m,1H),1.91-1.85(m,3H),1.68-1.52(m,
13
9H),1.27-1.23(m,49H),0.86(t,J=5.7Hz,J=6.6Hz,6H). CNMR(75MHz,DMSO-d6):δ/ppm = 172.22,171.96,171.83,170.85,170.40,170.29,168.19,168.12,65.27,64.80,
58.98,52.52,52.41,49.08,48.97,48.85,48.73,45.99,38.74,36.32,31.77,30.06,
29.55,29.49,29.44,29.18,28.50,28.47,26.80,26.23,25.80,25.73,23.95,22.88,
22.55,22.46,18.28,14.36.C,67.20%;H,10.67%;N,8.52%;O,13.62%.[0084] 实施例27制备Pro-Ala-Lys-Asp[OCH2(CH2)14CH3]-[OCH2(CH2)14CH3](4e)[0085] 按照实施例7的方法由265mg(0.25mmol)Boc-Pro-Ala-Lys(Boc)-Asp[OCH2(CH2
25
)14CH3]-[OCH2(CH2)14CH3]制得206mg(88%)目标化合物,为无色粉末。Mp 104℃;[α]D+
= -1116(c = 0.27,CHCl3);ESI-MS(m/e):878[M+H] ;IR(KBr):3300,3219,2957,2920,
1
2851,2758,2654,2575,1740,1643,1547,1468,1395,1371,1341,1292,1225,1169,1070.H NMR(300MHz,DMSO-d6):δ/ppm=10.11(s,1H),8.39(d,J=7.2Hz,1H),8.14(d,J=5.4Hz,
1H),4.62(s,1H),4.35(d,J = 6.6Hz,1H),4.25(s,2H),3.98-3.97(m,4H),3.22-3.19(m,
2H),2.73(s,4H),2.32-2.30(m,1H),1.87(m,3H),1.53(s,8H),1.22(s,55H),0.85(m,
13
6H). C NMR(75MHz,DMSO-d6):δ/ppm=171.84,171.74,171.67,170.78,170.24,168.21,
168.11,65.30,64.80,59.12,52.63,52.48,49.11,48.96,48.86,46.03,38.78,36.38,
36.26,31.86,31.71,29.99,29.66,29.45,29.42,29.37,29.10,28.47,26.77,25.77,
25.70,23.90,22.59,22.48,18.23,18.00,14.42,14.29,14.11.C,68.37%;H,10.90%;N,
7.97%;O,12.75%.
D = -1760(c = 0.01,CHCl3);ESI-MS(m/e):934[M+H] ;IR(KBr):3329,3275,3229,
1
3048,2922,2855,1740,1663,1545,1458,1240,1173.H NMR(300MHz,DMSO-d6):δ/ppm =
8.75(d,J=7.2Hz,1H),8.26(d,J=7.8Hz,1H),8.03-8.11(d,J=7.8Hz,1H),4.64(t,J=5.7Hz,J=6.0Hz,1H),4.38-4.33(m,1H),4.26-4.20(m,2H),4.04-3.95(m,4H),2.74(m,
4H),2.34-2.28(m,1H),1.92-1.86(m,3H),1.56(m,7H),1.37-1.25(m,68H),0.86(t,J =
13
6.0Hz,J = 6.6Hz,6H). C NMR(75MHz,DMSO-d6):δ/ppm = 171.85,171.65,170.78,
170.25,168.24,168.12,65.57,65.31,64.81,59.75,59.15,59.06,52.61,52.47,52.38,
52.28,49.21,49.03,48.86,46.38,46.30,46.19,46.07,38.95,36.46,31.86,31.71,
29.98,29.44,29.09,28.51,28.48,26.80,25.78,25.71,23.90,22.48,22.41,18.26,
15.73,14.29.C,69.41%;H,11.11%;N,7.49%;O,11.99%.