从昆虫体内分离抗循环疾病药物的先导化合物
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摘要
昆虫是地球生物圈中种类最多而且个体之间差别最大一群。从进化论的角度来看，它们和脊椎动物，植物，微生物有很大的差别。由于以上原因，研究昆虫发现新型生物活性分子的可能性很大。在本课题中，使用四种微量酶抑制剂活性检查系统和一种动物体内酶抑制剂活性检查系统搜索检查了110种昆虫的五种有机溶剂的550个萃取物的活性。最后，我们选择二种昆虫用于胆固醇酰基转移酶抑制剂的分离纯化课题，选择一种昆虫用于Lp-PLA_2酶抑制剂的分离纯化课题。在此课题中，我们采用活性指导的新的分离方法进行天然物的分离纯化。为了分离易于进行有机合成的小分子化合物，仪用了分子筛和树脂柱层析进行小分子化合物的筛选。
作为胆固醇酰基转移酶抑制剂，从M. phalerate, A. parki, 和P. brevitarsis三种昆虫的有机溶剂萃取物中分离纯化了八个已知化合物(1，2，4，5，6，7，8，和9)及两个新化合物(3，10)，其中化合物3，10，和6抑制小鼠肝脏微粒体胆固醇酰基转移酶，人体胆固醇酰基转移酶一型，人体胆固醇酰基转移酶二型的半数有效浓度分别是281，177，131μmol／l (3)，78，43，32μmol／l (10)，9.6，6.6，8.6μmol／l
(6)，作为Lp-PLA_2酶抑制剂，从P. cavimanus昆虫的有机溶剂萃取物中分离纯化了三个已知化合物(11，12，13)，化合物11，12，和13抑制Lp-PLA_2酶的半数有
效浓度分别是151，53，和45协mol/l。以上所有的化合物都进行了详细的谱图分
析，准确的判定了它们的结构。虽然在文献中对其中某些己知化合物的各种生
物活性有所涉及，但作为胆固醇酞基转移酶抑制剂或Lp一PL戈酶抑制剂还是第一
次.
Insects are the largest and most diverse group of organisms. They are evolutionarily distant relations with vertebrates, plants, and microbial origin. For this reasons, the study of insects promises to reveal novel bioactive compounds. Scores of insects' extracts have been screened in the five kinds' assay system. We select three kinds' insects in the ACAT assay system and one kind of insect in the Lp-PLAi assay system to isolate and purify. The activity guided of isolation methods as the new isolation methods was used.in this project. In order to isolate small molecular compounds (organic synthesis easily), molecular filter and sepadex column were used.
As inhibitors of acyl-CoA: cholesterol acyltransferase (ACAT), compounds (1, 2, 4, 5, 6, 7, 8, and 9) and two new compounds (3, 10) were isolated and purified from the solvent extracts of three insects, M. phalerate, A. parki, and P. brevitarsis, and were elucidated by their spectroscopic data analysis. Compounds 3, 10, and 6 inhibit rat liver microsomal ACAT, hACATl, and hACAT2 with IC50 values of 281, 177, 131 umol/i for 3, of 78, 43, 32 (amol/1 for 10, and of 9.6, 6.6, 8.6 |amol/l for 6, respectively. Three Lp-PLAa inhibitors (11, 12, and 13) were isolated and purified from the solvent extracte of the insect, P. cavimamis, and were elucidated by their spectroscopic data analysis.
Compounds 11, 12, and 13 inhibit Lp-PLA2 with ICso values of 151, 53, and 45 umol/1, respectively. Some of isolated compounds (1, 2, 4, 5, 6, 7, 8, 9, 11, 12, and 13) have been reported to show various biological activities, as potential inhibitors of human ACAT or human Lp-PLAz were first evaluated.
作为胆固醇酰基转移酶抑制剂，从M. phalerate, A. parki, 和P. brevitarsis三种昆虫的有机溶剂萃取物中分离纯化了八个已知化合物(1，2，4，5，6，7，8，和9)及两个新化合物(3，10)，其中化合物3，10，和6抑制小鼠肝脏微粒体胆固醇酰基转移酶，人体胆固醇酰基转移酶一型，人体胆固醇酰基转移酶二型的半数有效浓度分别是281，177，131μmol／l (3)，78，43，32μmol／l (10)，9.6，6.6，8.6μmol／l
(6)，作为Lp-PLA_2酶抑制剂，从P. cavimanus昆虫的有机溶剂萃取物中分离纯化了三个已知化合物(11，12，13)，化合物11，12，和13抑制Lp-PLA_2酶的半数有
效浓度分别是151，53，和45协mol/l。以上所有的化合物都进行了详细的谱图分
析，准确的判定了它们的结构。虽然在文献中对其中某些己知化合物的各种生
物活性有所涉及，但作为胆固醇酞基转移酶抑制剂或Lp一PL戈酶抑制剂还是第一
次.
Insects are the largest and most diverse group of organisms. They are evolutionarily distant relations with vertebrates, plants, and microbial origin. For this reasons, the study of insects promises to reveal novel bioactive compounds. Scores of insects' extracts have been screened in the five kinds' assay system. We select three kinds' insects in the ACAT assay system and one kind of insect in the Lp-PLAi assay system to isolate and purify. The activity guided of isolation methods as the new isolation methods was used.in this project. In order to isolate small molecular compounds (organic synthesis easily), molecular filter and sepadex column were used.
As inhibitors of acyl-CoA: cholesterol acyltransferase (ACAT), compounds (1, 2, 4, 5, 6, 7, 8, and 9) and two new compounds (3, 10) were isolated and purified from the solvent extracts of three insects, M. phalerate, A. parki, and P. brevitarsis, and were elucidated by their spectroscopic data analysis. Compounds 3, 10, and 6 inhibit rat liver microsomal ACAT, hACATl, and hACAT2 with IC50 values of 281, 177, 131 umol/i for 3, of 78, 43, 32 (amol/1 for 10, and of 9.6, 6.6, 8.6 |amol/l for 6, respectively. Three Lp-PLAa inhibitors (11, 12, and 13) were isolated and purified from the solvent extracte of the insect, P. cavimamis, and were elucidated by their spectroscopic data analysis.
Compounds 11, 12, and 13 inhibit Lp-PLA2 with ICso values of 151, 53, and 45 umol/1, respectively. Some of isolated compounds (1, 2, 4, 5, 6, 7, 8, 9, 11, 12, and 13) have been reported to show various biological activities, as potential inhibitors of human ACAT or human Lp-PLAz were first evaluated.
引文
1. Schoonjans, K.; Staels, B.; Auwerx, J. J. lipid Res. 1996, 37, 907.
2. Haugen, R and Norum, K. R. Scand. J. Gastroenterol. 1976, 11, 615.
3. Howles, P. N.; Carter, C. P.; Hui, D. Y. J. Biol. Chem. 1996, 271, 7196.
4. Goldstein, J. L.; Brown M. S. Nature 1990, 343, 425.
5. Sabine, J. R. Boca Raton FL. pp. 1-257.
6. H(?)rthle, K. Z. Physiol. Chem. 1895, 21, 332.
7. Windaus, A. Z. Physiol. Chem. 1910, 67, 174.
8. Sperry, W. M. J. Biol. Chem. 1935, 111, 467.
9. Glomset, J. A. J. Lipid Res. 1968, 9, 155.
10. D. S. Goodman, D. S.; Deykin, D.; Shiratori, T. J. Biol. Chem. 1964, 239, 1335.
11. Zhang, Y.; Yu, C.; Liu, J.; Chang, C. C. Y.; Chang, T. Y. J. Biol. Chem. 2003, 278, 11642.
12. Willner, E. L.; Tow, B.; Buhman, K. PNAS, 2003, 100, 1262.
13. Chao, H.; Zhou, M.; McIntosh, A.; J. Lipid Res. 2003, 44, 72.14. Doolittle, G. M; Chang, T. Y. Biochemistry. 1982, 21, 674.
15. Chang, C. C. Y.; Huh, H. Y.; Cadigan, K. M.; Chang, T. Y. J. Biol. Chem. 1993, 268, 20747.
16. Cadigan, K. M.; Heider, J. G.; Chang, T. Y. J. Biol. Chem. 1955, 263, 274.
17. Meiner, V.; Tam, C.; Gunn, M. D.; Dong, L. M. J. Lipid Res. 1997, 38, 1928.
18. Billheimer, J. T.; Cromley, D. A.; Kempner, E. S. J. Biol. Chem. 1990, 265, 8632.
19. Erickson, S. K.; Lear, S. R.; McCreery, M. J. J. Lipid Res. 1994, 35, 763.
20. Lin, S.; Cheng, D.; Chang, T.-Y. J. Biol. Chem. 1999, 274, 23276.
21. Cao, G.; Goldstein, J. L.; Brown, M. S. J. Biol. Chem. 1996, 271, 14642.
22. Lee, O.; Chang, C. C. Y.; Lee, W.; Chang, T.-Y. J. Lipid Res., 1995, 39, 1722.
23. Miyazaki, A.; Sakashita, N.; Lee, O. Arterioscler Thromb. Vasc. Biol., 1998, 18, 1568.
24. Anderson, R. A.; Joyce, C.; Davis, M. J. Biol. Chem. 1998, 273, 26747.
25. Cases, S.; Novak, S.; Zheng, Y.-W. J. Biol. Chem. 1995, 273, 26755.
26. Oelkers, P.; Behari, A.; Cromley, D. J. Biol. Chem. 1995, 273, 26765.
27. Lee, O.; Chang, C. C. Y.; Lee, W. J. Lipid Res. 1998, 39, 1722.
28. Chang, C. C. Y.; Lee, O.; Lin, S.; Chang, T.-Y. Circulation 1999, 100, 1-612.
29. Ross, R. Nature 1993, 362, 801.
30. Chang, T.Y.; Chang, C. C. Y.; Lin, S.; Yu, C. Curr. Opin. Lipidol. 2001, 12, 289.
31. Kim, Y. K.; Lee, H. W. J. Antibio. 1996, 49, 31.
32. Chang, T. Y.; Chang, C. C. Y.; Cheng, D. Anuu. Rev. Biochem. 1997, 66, 613.
33. (a) Krause, B. R.; Anderson, M.; Bisgaier, C. L.; Bocan, T.; Boustey, R.; Dehart, P.; Newton, R. S. J. Lipid. Res. 1993, 34, 279. (b) Carr, T. P.; Hamiltton, R. L.; Rudel, L. L. J. Lipid. Res. 1995, 36, 2 (c) Lee, H. T.; Sliskovic, D. R; Picard, J. A.; Roth, B. D.; Wierenga, W.; Hicks, J. J. Med. Chem. 1996, 39, 5031.
34. Burntt, J. R.; Wilcox, L. J.; Huff, M. W. Clin. Chimi. Acta. 1999, 286, 231.
35. Bocan, T. M. A.; Mueller, S. B.; Uhlendorf, E D. Arterioscler. Throm. 1991, 11, 1830.
36. Marzetta, C. A.; Savoy, Y. E. J. Lipid. Res. 1994, 35, 1829.
37. Ross, A. C.; Go, K. G.; Heider, J. G. J. Biol. Chem. 1984, 259, 815.
38. Drevon, C. A.; Engelhorn, S. C. and Steinberg, D. J. Lipid. Res. 1980, 21, 1065.
39. Murakami, S.; Araki, H.; Otomo, S. Life. Sci. 1995, 56, 509.
40. Matsuyama, N.; Kosaka, T.; Fukuhara, M. Bioorg. Med. Chem. Lett. 1999, 9, 2309.
41. Higley, C. A.; Wilde, R. G.; Maduskuie, T. E J. Med. Chem. 1994, 37, 3511.
42. Tanaka, A.; Tersawa, T.; Hagihara, H. J. Med. Chem. 1998, 41, 2390.
43. Chiari, A.; Lovisolo, P.; Radice, A. Pharmacol. Res. 1995, 32, 159.
44. Tawada, H.; Harcourt, M.; Kawamura, N. J. Med. Chem. 1994, 37, 2079.
45. Maduskuie, T. P.; Jr., Wilde, R. G.; Billheimer, J. T. J. Med. Chem. 1995, 38, 1067.
46. Li, H-Y.; Delucca, I.; Drummond, S. Heterocycles 1996, 43, 937.
47. Kuroda, K.; Yoshida, M.; J. Antibiot. 1993, 46, 1196.
48. Hasumi, K.; Shinohara, C.; Iwanaga, T. J. Antibiot. 1993, 46, 1303.
49. Schultz, T. H.; Lowe, R.; Woodby, R. A. Fed Proc. 1980, 39, 554.
50. Dixit, V. P.; Jain, P.; Bhandari, K. Indian J. Pharm. Sci. 1991, 53, 88.
51. Kwon, B. M.; Ro, S. H.; Kim, M. K. Planta. Med. 1997, 63, 552
52. Fukuda, T.; Kitada, Y.; Chen, X. M.; Chem. Pharm. Bull. 1996, 44, 2173.
53. Kwon, B. M.; Jung, H. J.; Lim, J. H. Planta. Med. 1999, 65, 74.
54. Ross, R.; N Engl J Med. 1999, 340, 115.
55. Jessup, W.; Kritharides, L. Curr Opin Lipidol. 2000, 11, 473.
56. Packard, C. J.; O'Reilly, D. S. J. et al. N Engl J Med. 2000, 343, 1148.
57. Maephee, C. H.; Moores, K. E; Boyd, H. F. Biochem J. 1999, 338, 479.
58. Caslake, M. J.; Packard, C. J.; Suckling, K. E Atherosclerosis 2000, 150, 413.
59. Nielson, L. B. Atherosclerosis 1999, 143, 229.
60. Tew, D. G.; Boyd, H. F.; Ashman, S. Biochemistry 1998, 37, 10087.
2. Haugen, R and Norum, K. R. Scand. J. Gastroenterol. 1976, 11, 615.
3. Howles, P. N.; Carter, C. P.; Hui, D. Y. J. Biol. Chem. 1996, 271, 7196.
4. Goldstein, J. L.; Brown M. S. Nature 1990, 343, 425.
5. Sabine, J. R. Boca Raton FL. pp. 1-257.
6. H(?)rthle, K. Z. Physiol. Chem. 1895, 21, 332.
7. Windaus, A. Z. Physiol. Chem. 1910, 67, 174.
8. Sperry, W. M. J. Biol. Chem. 1935, 111, 467.
9. Glomset, J. A. J. Lipid Res. 1968, 9, 155.
10. D. S. Goodman, D. S.; Deykin, D.; Shiratori, T. J. Biol. Chem. 1964, 239, 1335.
11. Zhang, Y.; Yu, C.; Liu, J.; Chang, C. C. Y.; Chang, T. Y. J. Biol. Chem. 2003, 278, 11642.
12. Willner, E. L.; Tow, B.; Buhman, K. PNAS, 2003, 100, 1262.
13. Chao, H.; Zhou, M.; McIntosh, A.; J. Lipid Res. 2003, 44, 72.14. Doolittle, G. M; Chang, T. Y. Biochemistry. 1982, 21, 674.
15. Chang, C. C. Y.; Huh, H. Y.; Cadigan, K. M.; Chang, T. Y. J. Biol. Chem. 1993, 268, 20747.
16. Cadigan, K. M.; Heider, J. G.; Chang, T. Y. J. Biol. Chem. 1955, 263, 274.
17. Meiner, V.; Tam, C.; Gunn, M. D.; Dong, L. M. J. Lipid Res. 1997, 38, 1928.
18. Billheimer, J. T.; Cromley, D. A.; Kempner, E. S. J. Biol. Chem. 1990, 265, 8632.
19. Erickson, S. K.; Lear, S. R.; McCreery, M. J. J. Lipid Res. 1994, 35, 763.
20. Lin, S.; Cheng, D.; Chang, T.-Y. J. Biol. Chem. 1999, 274, 23276.
21. Cao, G.; Goldstein, J. L.; Brown, M. S. J. Biol. Chem. 1996, 271, 14642.
22. Lee, O.; Chang, C. C. Y.; Lee, W.; Chang, T.-Y. J. Lipid Res., 1995, 39, 1722.
23. Miyazaki, A.; Sakashita, N.; Lee, O. Arterioscler Thromb. Vasc. Biol., 1998, 18, 1568.
24. Anderson, R. A.; Joyce, C.; Davis, M. J. Biol. Chem. 1998, 273, 26747.
25. Cases, S.; Novak, S.; Zheng, Y.-W. J. Biol. Chem. 1995, 273, 26755.
26. Oelkers, P.; Behari, A.; Cromley, D. J. Biol. Chem. 1995, 273, 26765.
27. Lee, O.; Chang, C. C. Y.; Lee, W. J. Lipid Res. 1998, 39, 1722.
28. Chang, C. C. Y.; Lee, O.; Lin, S.; Chang, T.-Y. Circulation 1999, 100, 1-612.
29. Ross, R. Nature 1993, 362, 801.
30. Chang, T.Y.; Chang, C. C. Y.; Lin, S.; Yu, C. Curr. Opin. Lipidol. 2001, 12, 289.
31. Kim, Y. K.; Lee, H. W. J. Antibio. 1996, 49, 31.
32. Chang, T. Y.; Chang, C. C. Y.; Cheng, D. Anuu. Rev. Biochem. 1997, 66, 613.
33. (a) Krause, B. R.; Anderson, M.; Bisgaier, C. L.; Bocan, T.; Boustey, R.; Dehart, P.; Newton, R. S. J. Lipid. Res. 1993, 34, 279. (b) Carr, T. P.; Hamiltton, R. L.; Rudel, L. L. J. Lipid. Res. 1995, 36, 2 (c) Lee, H. T.; Sliskovic, D. R; Picard, J. A.; Roth, B. D.; Wierenga, W.; Hicks, J. J. Med. Chem. 1996, 39, 5031.
34. Burntt, J. R.; Wilcox, L. J.; Huff, M. W. Clin. Chimi. Acta. 1999, 286, 231.
35. Bocan, T. M. A.; Mueller, S. B.; Uhlendorf, E D. Arterioscler. Throm. 1991, 11, 1830.
36. Marzetta, C. A.; Savoy, Y. E. J. Lipid. Res. 1994, 35, 1829.
37. Ross, A. C.; Go, K. G.; Heider, J. G. J. Biol. Chem. 1984, 259, 815.
38. Drevon, C. A.; Engelhorn, S. C. and Steinberg, D. J. Lipid. Res. 1980, 21, 1065.
39. Murakami, S.; Araki, H.; Otomo, S. Life. Sci. 1995, 56, 509.
40. Matsuyama, N.; Kosaka, T.; Fukuhara, M. Bioorg. Med. Chem. Lett. 1999, 9, 2309.
41. Higley, C. A.; Wilde, R. G.; Maduskuie, T. E J. Med. Chem. 1994, 37, 3511.
42. Tanaka, A.; Tersawa, T.; Hagihara, H. J. Med. Chem. 1998, 41, 2390.
43. Chiari, A.; Lovisolo, P.; Radice, A. Pharmacol. Res. 1995, 32, 159.
44. Tawada, H.; Harcourt, M.; Kawamura, N. J. Med. Chem. 1994, 37, 2079.
45. Maduskuie, T. P.; Jr., Wilde, R. G.; Billheimer, J. T. J. Med. Chem. 1995, 38, 1067.
46. Li, H-Y.; Delucca, I.; Drummond, S. Heterocycles 1996, 43, 937.
47. Kuroda, K.; Yoshida, M.; J. Antibiot. 1993, 46, 1196.
48. Hasumi, K.; Shinohara, C.; Iwanaga, T. J. Antibiot. 1993, 46, 1303.
49. Schultz, T. H.; Lowe, R.; Woodby, R. A. Fed Proc. 1980, 39, 554.
50. Dixit, V. P.; Jain, P.; Bhandari, K. Indian J. Pharm. Sci. 1991, 53, 88.
51. Kwon, B. M.; Ro, S. H.; Kim, M. K. Planta. Med. 1997, 63, 552
52. Fukuda, T.; Kitada, Y.; Chen, X. M.; Chem. Pharm. Bull. 1996, 44, 2173.
53. Kwon, B. M.; Jung, H. J.; Lim, J. H. Planta. Med. 1999, 65, 74.
54. Ross, R.; N Engl J Med. 1999, 340, 115.
55. Jessup, W.; Kritharides, L. Curr Opin Lipidol. 2000, 11, 473.
56. Packard, C. J.; O'Reilly, D. S. J. et al. N Engl J Med. 2000, 343, 1148.
57. Maephee, C. H.; Moores, K. E; Boyd, H. F. Biochem J. 1999, 338, 479.
58. Caslake, M. J.; Packard, C. J.; Suckling, K. E Atherosclerosis 2000, 150, 413.
59. Nielson, L. B. Atherosclerosis 1999, 143, 229.
60. Tew, D. G.; Boyd, H. F.; Ashman, S. Biochemistry 1998, 37, 10087.