紫苏叶抗高尿酸血症有效部位及其制备方法和应用_3

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Hz, H-5),6. 66 (lH,d,/= 1. 5Hz,H-2'),6. 58 (lH,d,/= 8. 0Hz,H-5'),6. 47 (lH,dd,8. 0, 1. 6Hz,H-6'),6. 16 (1H,d,/= 15. 9Hz,H-8),4. 83 (1H,dd,/= 10. 0,3. 0Hz,H-8'), 2. 99 (1H,dd,/ = 14. 2, 3. 0Hz,H-7' ),2. 74 (1H,dd,/ = 14. 2,10. 0Hz,H-7' 々); 13C-NMR(125MHz,DMS0_4) 5:172.3 (C-9'),166.3 (C-9),148.7 (C-4),146.1 (C-3), 144.9 (C-3,),144.2 (C-7),143.5 (C-4,),130.0 (C-l,),125.4 (C-l),120.5 (C-6), 119. 5 (C-6,),116. 6 (C-2,),115. 9 (C-5),115. 4 (C-5,),115. 1 (C-2),114. 7 (C-8),76. 2 (C-8,),37. 3 (C-7,);ESI-MSa?/z:359 [M-H]。
[0039]化合物 4 ^H-NMR(500MHz,CD30D) 5:7. 56 (lH,d,/= 15. 9Hz,H-7),7. 05 (1H, d,/= 2.0Hz,H-2),6.96 (lH,dd,/= 8.2,2.0Hz,H-6),6.78 (lH,d,/= 8.2Hz,H-5), 6.71 (lH,d,/= 2.0Hz,H-2'),6.70 (lH,d,/= 8.2Hz,H-5'),6.57 (lH,dd,/= 8.0, 2. 0Hz,H-6'),6. 26 (1H,d,/= 15. 8Hz,H-8),5. 20 (1H,dd,/= 10. 4,5. 2Hz,H-8'), 3.70 (3H,s,OCH3),3. 04 (2H,m,H-7');13C-NMR(125MHz,CD30D) 5:172.2 (C-9,),168. 3 (C-9),149. 8 (C-4),148. 0 (C-7),146. 8 (C-3),146. 2 (C-3'),145. 4 (C-4'),128. 8 (C-1'), 127. 6 (C-l),123. 2 (C-6),121. 8 (C-6,),117. 6 (C-2,),116. 5 (C-5),116. 3 (C-5,),115. 3 (C-2),114.2 (C-8),74.7 (C-8'),52.7 (_OCH3),37.9 (C-7');ESI-MS聲々:374[M]+。
[0040] 化合物 5 ^H-NMR(500MHz,CD30D) 5:7. 59 (lH,d,/= 15. 6Hz,H-7),7. 44 (2H, d,/= 8.4Hz,H-2,6),6.80 (lH,d,/= 8.5Hz,H-3,5),6.28 (lH,d,/= 15.8Hz,H-8); 13C-NMR(125MHz,CD30D)H71.2 (C-9),161.1 (C-4),146.5 (C-7),131.0 (C-2,6), 127.3 (C-l),116.8 (C-3,5),116.1 (C-8);ESI-MSa?/z:163 [M-H]。
[0041] 化合物 6 ^H-NMR(500MHz,DMS0) 5:12. 72 (1H,s,5-0H),10. 36 (1H,s,6-0H), 8.58 (lH,s,4'-0H),7.91 (2H,d,/=8.8Hz,H-2',6'),6.98 (lH,s,H-8),6.93 (2H,d,/ =8.8Hz,H-3',5'),6.79 (lH,s,H-3),5. 19 (lH,d,/= 7.4Hz,H-l''),4.01 (lH,d,/ = 9.6Hz,H-5,'),3. 38 (3H,m,H-2,,,3,,,4,');13C-NMR(125MHz,DMS0)A182.3 (C-4), 170. 1 (C-6"),164. 1 (C-7),161. 2 (C-2),151. 0 (C-9),149. 0 (C-4,),146. 8 (C-5),130. 5 (C-6),128.4 (C-2',6,),121.2 (C-l,),116.0 (C-3',5,),105.8 (C-10),102.5 (C-3), 100.1 (C-1"),93.6 (C-8),75.4 (C-5"),75.2 (C-3"),72.8 (C-2"),71.3 (C-4");ESI-MS a?/z:463 [M+H] +〇
[0042] 化合物 7 ^H-NMR(DMS0_4,500MHz)A9. 50 (lH,s,4,-0H),8. 01 (lH,s,8-0H), 7. 31 (2H,d,/= 8. 6Hz,H-2',6'),6. 76 (2H,d,/= 8. 6Hz,H-3',5'),6. 31 (lH,s,H-6), 5. 35 (lH,dd,/= 12. 0,3. 0Hz,H-2),3. 86 (3H,s,7-0CH3),3. 75 (3H,s,5-0CH3),2. 99 (1H, dd,/= 16.4,12.0Hz,H-3々),2.60 (lH,dd,/= 16.4,3.1Hz,H-3ff);13C-NMR(DMSO-4, 125MHz)A188.6 (C-4),157.4 (C-4,),153.8 (C-5),153.6 (C-7),150.7 (C-9),129. 3 (C-1'),128. 0 (C-2,,6'),127. 8 (C-8),115. 0 (C-2,,5'),105. 7 (C-10),90. 4 (C-6),78. 1 (C-2),55.9 (5,7-0CH3),44.9 (C-3);ESI-MS餘々:339 [M+Na] +。
[0043] 化合物 8 ^H-NMR(500MHz,DMS0_4) 5:12. 50 (lH,brs,5-0H),8. 73 (lH,brs, 6-0H),8. 09 (2H,dd,/= 7. 8,1. 1Hz,H-2',6'),7. 58 (3H,m,H-3',4',5'),6. 98 (1H,s, H-3),6.95 (lH,s,H-8),3.92 (3H,s,-0CH3);13C-NMR(125MHz,DMS0_4)A182.3 (C-4), 163. 1 (C-2),154. 6 (C-7),149. 8 (C-9),146. 1 (C-5),131. 9 (C-4,),130. 8 (C-1'),130. 1 (C-6),129.1 (C-3',5,),126.3 (C-2',6,),105.3 (C-10),104.7 (C-3),91. 3 (C-8),56. 3 (_0CH3) ;ESI-MSa?/z:285 [M+H] +。
[0044] 化合物 9 ^H-NMR(500MHz,DMSO-4) 5:12. 95 (1H,s,5-0H),7. 40 (1H,dd,/ = 8. 1,2. 2Hz,H-6'),7. 38 (lH,d,/= 2. 2Hz,H-2'),6. 87 (lH,d,/= 8. 1Hz,H-5'),6. 62 (lH,s,H-3),6.42 (lH,d,/= 1.8Hz,H-8),6.16 (lH,d,/= 1.9Hz,H-6);13C-NMR(125 MHz,DMS0_4))A181.5 (C-4),164.5 (C-7),163.8 (C-2),161.4 (C-5),157.3 (C-9), 150. 0 (C-4,),145. 8 (C-3,),121. 3 (C-6,),118. 9 (C-1'),116. 0 (C-5,),113. 2 (C-2,), 103.5 (C-10),102.7 (C-3),98.9 (C-6),93.8 (C-8);ESI-MS餘285 [M-H]。
[0045] 化合物 10 ^H-NMR(500MHz,DMSO-4) 5:12. 92 (1H,s,5-0H),7. 88 (2H,d,/ = 8. 7Hz,H-2',6'),6. 91 (2H,d,/= 8. 7Hz,H-3',5'),6. 70 (lH,s,H-3),6. 41 (lH,brs, H-8),6. 13 (1H,brs,H-6);13C-NMR(125MHz,DMSO-4)A181.4 (C-4),165.4 (C-7), 163.5 (C-2),161.3 (C-5),161.3 (C-4,),157.4 (C-9),128.3 (C-2',6,),121.0 (C-l,), 116.0 (C-3,,5'),103.2 (C-10),102.6 (C-3),99.2 (C-6),94.0 (C-8);ESI-MS聲々:270 [M]+〇
[0046] 实施例8 紫苏叶抗高尿酸血症有效部位黄嘌呤氧化酶抑制活性测定 HPLC测定方法:Diamonsil0DS色谱柱(250mmX4.6mm,5jwm);流动相:100mmol/L磷酸二氢钠溶液(pH=3. 5);检测波长:290nm;流速:1. 0mL/min。
[0047] 测定体系:不同浓度测试品溶液(含0. 25%DMS0的0. 07mol/L磷酸盐缓冲液)200 加入0. 07mol/L磷酸盐缓冲液140 #L,黄嘌呤氧化酶溶液120 (0. 02unit/mL, 0. 07mol/L磷酸盐缓冲液),25°C保温15min,加入黄嘌呤溶液240 #L(300 #m〇l/L,0. 07mol/L磷酸盐缓冲液),25°C保温15min,加入1mol/L盐酸100 终止反应。空白对照测 定,供试品溶液用含1 %DMS0的0.07mol/L磷酸盐缓冲液代替。HPLC法测定尿酸生成量, 每组浓度平行三次,计算抑制率。结果见表2。
[0048] 表2紫苏叶抗高尿酸血症有效部位对黄嘌呤氧化酶抑制率
实施例9 紫苏叶抗高尿酸血症有效部位对实验性高尿酸血症模型小鼠血清尿酸水平的影响 小鼠随机分为6组,每组20只,分别为正常对照组、模型对照组,高剂量组(实施例3制 备的紫苏叶抗高尿酸血症有效部位1g/Kg),中剂量组(实施例3制备的紫苏叶抗高尿酸血 症有效部位〇. 5g/Kg),低剂量组(实施例3制备的紫苏叶抗高尿酸血症有效部位0. 25
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