δΗ 1.04,1. 06(d,each 3H, J = 6. 5Hz,HIken5i6), L H(s, 3H, H-19), L 24, I. 26, L 33(d, each 3H, J = 6. 0Hz, H-6 of sugar moieties), I. 40 (s, 3H, H-18), 2. 11 (s, 3H, Hlken 7), 2. 16 (s, 3H, H-21), 3. 38, 3 .45 (s, each 3H, 3-〇CH3 of sugar moieties), 4. 49 (dd, 1H, J = 2. 0, 9. 6 Hz, anomeric H), 4. 81 (dd, 1H, J = 2.2,9.3 Hz, anomeric H), 4. 92 (dd, 1H, J = 2.2,9.6 Hz, anomeric H),5. 35 (brs, 1H, H-6),5. 51 (br s, 1H, Hlken 2) ;13C NMR(O)Cl3, 125 MHz) : δ c 38. 9 (t, C-l), 29.0(t,C-2),78.0(d,C-3),38.9(t,C-4),140.8(s,C-5),117.7(d,C-6),34.3(t,C-7),74. 4(s,C-8),43. 8(d,C-9),37. 3(s,C-10),24. 4(t,C-ll),71. 7(d,C-12),58. 0(s,C-13),88. I (s,C-14),33.2(t,C-15),32.0(t,C-16),91.6(s,C-17),9.5(q,C-18),18.7(q,C-19),209. 0 (s, C-20), 27. 3 (q, C-21), 166. 0 (s, Clken ^ , 113. I (d, Clken 2), 166. I (s, Clken 3), 38. 3 (s, Clken 4), 20. 9 (q, Clkem 5), 21. 0 (q, Clkem 6), 16. 7 (q, Clkem 7), 95. 9 (d, Cdigit λ), 37. 2 (t, Cdigit 2), 75. 5 (d ,C1Jigit 3 ),82.6(d, Cdlglt 4), 68. 2 (d, Cdigit 5) J 18· 3 (q, ?digit 6),98. 5 (d, Ccym D,35. 7 (t, Ccym 2),7 7. 0 (d,Ccym 3),82. 5 (d,Ccym 4),68. 8 (d,Ccym 5),18. I (q,Ccym 6),58. 5 (q,Ccym 0Me),101. 6 (d,Cole 1 ),35. 4 (t,Cole 2),80. 7 (d,Cole 3),75. 5 (d,Cole 4),71. 6 (d,Cole 5),18. I (q,Cole 6),56. 4 (q,Cole 0Me) 0
[0203] 化合物青陽參苷己對戊四唑(Pentylenetetrazole)誘導(dǎo)斑馬魚癲癇發(fā)作模型的 保護(hù)作用:
[0204] 按照實(shí)施例1中的方法對化合物的抗癲癇治療效果進(jìn)行定性評價(jià)。由圖5A可知, 在青陽參苷己各給藥劑量下與癲癇模型組相比,斑馬魚快速運(yùn)動(dòng)軌跡明顯減少。圖5B為根 據(jù)斑馬魚快速運(yùn)動(dòng)距離平均值mean ± SE繪制的柱形圖,由圖5B可知,化合物青陽參苷己在 30、100 μ M濃度下,有癲癇治療趨勢,但無統(tǒng)計(jì)學(xué)差異;在300 μ M濃度下有抗癲癇作用,癲 癇治療效率為55% (Ρ〈0. 01),量效關(guān)系明確。
[0205] 實(shí)施例6
[0206] 化合物羅素他命3-0-β -D-夾竹桃糖吡喃糖基-(1 - 4)-β -D-加拿大麻糖吡喃 基- (1 - 4)-β -D-加拿大麻糖[rostratamine 3-0-β -D-oleandropyranosyl-(1 - 4)-β -D-cymaropyranosyl-(l - 4)-β -D-cymaropyranoside](式 II)(以下簡稱"化合物 II") 體內(nèi)抗癲癇作用的評價(jià):
[0207] 化合物II的制備方法:干燥的青陽參塊根5kg,粉碎用95%乙醇滲濾提取三次, 每次2~3天,合并提取液,減壓濃縮得到青陽參的醇浸膏,將提取物以水混懸,乙酸乙酯萃 取,得乙酸乙酯萃取物,C-18反相柱色譜以甲醇的水溶液(10%~100% )梯度洗脫,得到 組分Frl-Fr5, Fr4進(jìn)一步用制備HPLC反復(fù)純化得到純化合物II 100mg。
[0209] rostratamine3-〇_ β -D-oleandropyranosyl- (1 - 4) - β -D-cymaropyranosyl- (1 -4)-β -D-cymaropyranoside
[0210] 化合物II的理化數(shù)據(jù)如下:白色針狀結(jié)晶,易溶于甲醇,乙醇,丙酮,氯仿等有機(jī) 溶劑,分子式為〇 ;3!19602。,1!1匪1?(0)(:13,400 1抱):5"1.46(8,3!1,!1-18),1.20,1.22,1.31((1 ,each 3H, J = 6. 3 Hz, H-6 of sugar moiety), I. 12 (s, 3H, 19-CH3), 2. 19 (s, 3H, H-21), 3. 38 ,3. 44, 3. 44 (s, each 3H, 3-〇CH3 of sugar moieties), 4. 48 (dd, 1H, J = I. 9, 9. 8, anomeric H),4. 74(dd, J = I. 9, 9. 8, 1H, anomeric H), 4. 84 (dd, 1H, J = 1.8,9.6 Hz, anomeric H), 5. 37 (brs, 1H, H-6), 6. 30 (d, 1H, J = 16. 0 Hz, Hcin 2), 7. 38 (m, 3H, Hcin 6j7j8) , 7. 51 (m, 2H, H Cin 5i9), 7. 62(d, 1H, J=16. 0 Hz, Hcin 3) ;13C NMR(Pyridine-d5, 125 MHz): 5C 39. 9(t, C-l), 29. 8 (t, C-2), 77. 6 (d, C-3), 38. 8 (t, C-4), 139. 3 (s, C-5), 119. 2 (d, C-6), 34. 6 (t, C-7), 74. 2 (s, C-8),44. 5 (d, C-9),37. I (s, C-10),25. 2 (t, C-ll),73. 9 (d, C-12),58. 3 (s, C-13),89. 5 (s,C-14),34. 7 (t,C-15),33. 6 (t,C-16),92. I (s,C-17),10. 7 (q,C-18),18. I (q,C-19),209 ? 9 (s,C-20),27. 7 (q,C-21),165. 9 (s,Ccin J,119. I (d,Ccin 2),144. 7 (d,Ccin 3),135. 0 (s,Ccin 4), 128. 6 (d, Ccin 5i 9), 129. 3 (d, Ccin 6_8), 130. 6 (d, Ccin 7), 96. 3 (d, Ccyml λ), 37. 9 (t, Ccyml 2), 77. 8 (d, Ccyml 3), 83. 4 (d, Ccyml 4), 68. 7 (d, Ccyml 5), 18. 3 (q, Ccyml 6), 58. 6 (q, C), 100. 5 (d, Ccym2 i), 3 7. 2 (t, Ccym2 2), 78. 0 (d, Ccym2 3), 83. 2 (d, Ccym2 4), 68. 9 (d, Ccym2 5), 18. 5 (q, Ccym2 6), 58. 8 (q, Ccym2 0Me), 102. 2 (d, Cole ^ , 37. 2 (d, Cole 2), 81. 3 (d, Cole 3), 76. 2 (d, Cole 4), 72. 8 (d, Cole 5), 18. 6 (d, C ole 6),57. I (d, C〇ie 〇Me),
[0211] 化合物II對戊四唑(Pentylenetetrazole)誘導(dǎo)斑馬魚癲癇發(fā)作模型的保護(hù)作 用:
[0212] 按照實(shí)施例1中的方法對化合物的抗癲癇治療效果進(jìn)行定性評價(jià)。由圖6A可知, 在化合物II各給藥劑量下與癲癇模型組相比,斑馬魚快速運(yùn)動(dòng)軌跡明顯減少。圖6B為根 據(jù)斑馬魚快速運(yùn)動(dòng)距離平均值mean±SE繪制的柱形圖,由圖6B可知,化合物II在30 μ M 濃度下,有癲癇治療趨勢,但無統(tǒng)計(jì)學(xué)差異;在100、200 μ M濃度下均有抗癲癇作用,治療效 率分別為 65%、52% (Ρ〈0·01、Ρ〈0·01)。
[0213] 實(shí)施例7
[0214] 化合物wilfoside C3N(式VII)體內(nèi)抗癲癇作用的評價(jià):
[0215] Wilfoside C3N的制備方法:干燥的耳葉牛皮消塊根10kg,粉碎用95%乙醇滲濾 提取三次,每次2~3天,合并提取液,減壓濃縮得到耳葉牛皮消的醇浸膏,將提取物以水混 懸,乙酸乙酯萃取,得乙酸乙酯萃取物,將乙酸乙酯浸膏溶于70%乙醇溶液中,過濾或離心 去除不溶物后,進(jìn)行大孔樹脂柱色譜分離,收集75%乙醇洗脫流分,濃縮后得活性部位,活 性部位進(jìn)一步用制備HPLC反復(fù)純化得到純Wilfoside C3N。
[0217] Wilfoside C3N的理化數(shù)據(jù)如下:白色無定形粉末,易溶于甲醇,乙醇,丙酮,氯 仿等有機(jī)溶劑,分子式為 C49H78O16ZH NMR(Pyridine-d5, 400 MHz) : δ η 0· 92, 0· 94 (d,each 3H, J = 6. 5Hz,HIken5i6), L 32(s, 3H, H-19), L 36, I. 34, L 51(d, each 3H, J = 6. 0Hz, H-6 of sugar moieties), I. 97 (s, 3H, H-18), 2. 26 (s, 3H, Hlkem 7), 2. 50 (s, 3H, H-21), 3. 42, 3. 44, 3. 51 (s, each 3H, 3_0CH3 of sugar moieties), 5. 24 (dd, 1H, J = I. 9, 9. 4, anomeric H), 5. 15 (d, J = 3. 3, 1H, anomeric H), 5. 10 (dd, 1H, J = I. 8, 9. 8 Hz, anomeric H), 5. 86 (br s, 1H, Hlken 2) ;13C NMR(Pyridine-d5, 125 MHz) : δ c 39. 0 (t, C-l), 29. 9 (t, C-2), 77. 5 (d, C- 3),39. 3 (t, C-4),139. 3 (s, C-5),119. 2 (d, C-6),34. 8 (t, C-7),74. 3 (s, C-8),44. 6 (d, C-9) ,37. 4(s,C-10),25. I (t,C-ll),72. 6(d,C-12),58. 0(s,C-13),89. 5(s,C-14),33. 9(t,C-I 5),33. 0 (t,C-16),92. 4 (s,C-17),10. 8 (q,C-18),18. 2 (q,C-19),209. 5 (s,C-20),27. 6 (q, C_21), 166. 0 (s, Clkem j), 114. I (d, Clkem 2), 165. 5 (s, Clkem 3), 38. 2 (s, Clkem 4), 20. 9 (q, Clkem 5), 20. 9 (q, Clkem 6), 16. 5 (q, Clkem 7), 96. I (d, Ccyml ,) , 35. 3 (t, Ccyml 2), 77. 5 (d, Ccyml 3), 82. 5 (d, Ccy ml 4), 69. 2 (d, Ccyml 5), 18. 8 (q, Ccyml 6), 57. 2 (q, C), 101. 0 (d, Cdigin λ), 32. 5 (t, Cdigin 2), 74. 5 (d ,Cdigin 3),73. 8 (d, Cdigin 4),67. 5 (d, Cdigin 5),17. 9 (q, Cdigin 6),55. 3 (q, Cdigin 〇Me),99. 5 (d, Ccym2 I ),35. 3 (d, Ccym2 2)) 79. 9 (d, Ccym2 3), 74. I (d, Ccym2 4), 71. 0 (d, Ccym2 5), 18. 7 (d, Ccym2 6), 58. 0 (d, Ccym2 OMe) ·
[0218] Wilfoside C3N對戊四唑(Pentylenetetrazole)誘導(dǎo)斑馬魚癲癇發(fā)作模型的保 護(hù)作用:
[0219] 按照實(shí)施例1中的方法對化合物的抗癲癇治療效果進(jìn)行定性評價(jià)。由圖7A可知, 在wilfoside C3N各給藥劑量下與癲癇模型組相比,斑馬魚快速運(yùn)動(dòng)軌跡明顯減少。圖7B 為根據(jù)斑馬魚快速運(yùn)動(dòng)距離平均值mean土SE繪制的柱形圖,由圖7B可知,wilfoside C3N 在100 μ M濃度下,有癲癇治療趨勢,但無統(tǒng)計(jì)學(xué)差異;在200 μ M濃度下有抗癲癇作用,治療 效率為 55. 2% (Ρ〈0· 001)。
[0220] 實(shí)施例8
[0221] 化合物cynauricoside A(式VIII)體內(nèi)抗癲癇作用的評價(jià):
[0222] Cynauricoside A的制備方法:干燥的耳葉牛皮消塊根IOkg,粉碎用95%乙醇滲 濾提取三次,每次2~3天,合并提取液,減壓濃縮得到青陽參的醇浸膏,將提取物以水混 懸,乙酸乙酯萃取,得乙酸乙酯萃取物,將乙酸乙酯浸膏溶于70%乙醇溶液中,過濾或離心 去除不溶物后,進(jìn)行大孔樹脂柱色譜分離,收集75%乙醇洗脫流分,濃縮后得活性部位,活 性部位進(jìn)一步用制備HPLC反復(fù)純化得到純cynauricoside A。