新型过氧化物酶体增殖因子活化受体激动剂的设计、合成和构效关系的研究
摘要
2型糖尿病是一种代谢性疾病，对人体的危害仅次于癌症，其患者伴有胰岛素耐受，高血糖，高血脂等症状。临床治疗药物主要分三大类，即磺脲类、双胍类、α-糖苷酶抑制剂。噻唑烷二酮类(TZD_S)胰岛素增敏剂及其作用靶点过氧化物酶体增殖因子活化受体(PPAR)的发现开辟了一条治疗2型糖尿病的新道路。目前已知PPAR分三种亚型PPARα，PPARγ和PPARδ。现已有三个PPARγ激动剂(曲格列酮，罗格列酮和吡格列酮)作为2型糖尿病治疗药物先后上市。但是临床应用显示它们在治疗糖尿病、降低血糖的同时，会导致肥胖、心血管疾病等副作用。研究得知降脂药物如氯贝特的作用机理是由于激动PPARα，而产生降低血脂和减少肥胖的作用。因此，新的研究策略是将PPARγ激动剂的胰岛素增敏活性、糖调节活性和PPARα的脂调节活性结合起来，这样能同时激动PPARγ和PPARα的分子应是产生较少副作用的新一代2型糖尿病的治疗药。
为了寻找更加安全有效的2型糖尿病的治疗药，本课题进行了以下几个方面的工作：
1．对已知的PPAR激动剂进行了构效关系(SAR)研究，并将基于受体结构的药物分子设计与组合化学策略相结合，构建了过氧化物酶体增殖因子活化受体(PPAR)激动剂的虚拟化合物库。经分子对接和虚拟筛选，得到理论上结合较强的化合物，选取合适的构建模块，将其组合，并对所组合的化合物进行合成。共设计合成了六类化合物，噁唑酮类化合物(第Ⅰ类)，哌嗪-2，5-二酮类化合物(第Ⅱ类)，N-取代氨基酸类化合物(第Ⅲ类)，α-烷氧丙酸类化合物(第Ⅳ类)，丙二酸酯类化合物(第Ⅴ类)，甲磺酰胺类化合物(第Ⅴ类)，期望从中找出具降糖活性的先导化合物。
X=H，N
R1=H，-C_2H_5
R2=-CH_2OH，-CH_2COOH
R3=-CH_3，-C_2H_5，-iC_3H_7，-C_5H_9，-C_6H_(11)，-C_6H_(5…)
2．本论文共合成化合物145个，新化合物90个，目标化合物85个，全部经~1HNMR和MS的确证，其中24个化合物经高分辨质谱验证。并选取目标化合物FJ-07-17进行X-单晶衍射分析，得到晶体结构的数据。
3．利用已建立的药理模型对所合成的化合物进行生物活性评价，并对构效关系做了定性分析。鉴定PPAR激动剂生物活性的方法主要分为两部分，即分别测定分子的结合活性和功能活性。对所合成的53个化合物测定了化合物与PPARγ的结合能力；对所合成的82个化合物在10~(-5)M测定了转染PPARγ基因的细胞水平上的转录激活实验；然后从中选取活性较好的24个化合物，在六个浓度测定了转染PPARα，PPARγ基因的细胞水平上的转录激活实验。实验结果显示：α-烷氧丙酸类化合物FJ-08-46的活性较好，可以此化合物为先导物进行结构改造，期望获得高效、低毒的2型糖尿病的治疗药。目前化合物FJ-08-46体内药效学评价正在进行中。
4．PPARγ激动剂有抑制肿瘤生长和预防治疗癌症的作用，对所合成目标化合物进行了体外MTT法和集落形成法抑制肿瘤细胞生长实验和体内肿瘤模型裸鼠的动物实验。实验结果显示化合物FJ-05-05对结肠癌HT-29模型裸鼠肿瘤有一定抑制作用，可根据此模型进行筛选，期望找到高效低毒的抗癌药物。
Type 2 diabetes is a metabolic disease characterized by insulin resistance, hyperglycaemia, and hyperlipidemia, leading to impaired secretion of insulin in the later stsges. The disease is often associated with obesity, dyslipidemia, and hypertension , causing increased cardiovascular risks. Discovery of the insulin-sensitizing drugs thiazolidinedione(TZD) and the drug-target (Peroxisome proliferator-activated receptors, PPARs) offers a new route for the treatment of type 2 diabetes. PPARs are members of a huge nuclear hormone receptor superfamily, a group of nuclear proteins that mediate the specific effects of small lipophhilic compounds, such as steroids, retinoids, and fatty acids, on DNA transcription. Three subtypes of PPARs have been identified and cloned from mouse and human: PPARα, PPARγ, and PPARδ. To date three PPARγagonists Troglitazone, Rosiglitazone and Pioglitazone have been launched, which, however, provide minor protection against eventual cardiovascular risks that develop with type 2 diabetes. A new strategy of designing novel anti-diabetics is to combine the insulin sensitizing and glucose regulating activity of a PPARγagonists with lipid/cholesterol modulating properties of PPARαagonists (fibrate) into a single molecule. Dual-acting PPARαand PPARγagonists, designed to combine the beneficial effects seen with insulin sensitizers and fibrate, could be new drugs to 2 type diabetes.In order to find new and more effective drugs for the type 2 diatebes, the following work had been carried out in the thesis:1. Scrutinizing the structures of PPAR agonists and the binding feature of agonists to the receptors, a novel virtual library of PPAR agonists was constructed by structure-based drug design and combinatorial library method. Virtual screening was conducted by DOCKoperation, which automatically simulated the interaction between the ligands and receptors. Those strong-binding virtual compounds were selected according to the scoring functions. Six series of compounds composed of diverse building blocks were designed and synthesized in order to search for lead compounds with dual activity.
X=H, N
R_1=H, -C_2H_5
R2=-CH_2OH, -CH_2COOH
R3=-CH_3, -C_2H_5, -iC_3H_7, -C_5H_9, -C_6H_(11), -C_6H_(5…)
2. One hundred and forty-five chemical substances including ninety new compounds were prepared, of which eighty-five novel compounds are target molecules. All the targets moleculars were identified by ~1HNMR (300MHz) and MS (EI, FAB, or ESI), twenty-four of which were identified by high revolution mass spectroscopy. The compound FJ-07-17 was also identified by X-ray crystallography.
3. Most of the target compounds were evaluated in vitro. Some of the compounds evaluated show strong binding affinities to PPARγ. Some of the compounds were found possessing approximate activity to Pioglitazone in vitro PPARαand PPARγtransactivation. The Structure-activity relationships of synthsized compounds and transactivation data offered detailed molecular information for design and synthesis of new compounds. The evaluation of compound FJ-08-46 in vivo is in progresss.
4. Some compounds were in vitro tested for the activity on cell differentiation and proliferation. The compound FJ-05-05 was found to exhibit moderate inhibition of HT-29 cell line.
为了寻找更加安全有效的2型糖尿病的治疗药，本课题进行了以下几个方面的工作：
1．对已知的PPAR激动剂进行了构效关系(SAR)研究，并将基于受体结构的药物分子设计与组合化学策略相结合，构建了过氧化物酶体增殖因子活化受体(PPAR)激动剂的虚拟化合物库。经分子对接和虚拟筛选，得到理论上结合较强的化合物，选取合适的构建模块，将其组合，并对所组合的化合物进行合成。共设计合成了六类化合物，噁唑酮类化合物(第Ⅰ类)，哌嗪-2，5-二酮类化合物(第Ⅱ类)，N-取代氨基酸类化合物(第Ⅲ类)，α-烷氧丙酸类化合物(第Ⅳ类)，丙二酸酯类化合物(第Ⅴ类)，甲磺酰胺类化合物(第Ⅴ类)，期望从中找出具降糖活性的先导化合物。
X=H，N
R1=H，-C_2H_5
R2=-CH_2OH，-CH_2COOH
R3=-CH_3，-C_2H_5，-iC_3H_7，-C_5H_9，-C_6H_(11)，-C_6H_(5…)
2．本论文共合成化合物145个，新化合物90个，目标化合物85个，全部经~1HNMR和MS的确证，其中24个化合物经高分辨质谱验证。并选取目标化合物FJ-07-17进行X-单晶衍射分析，得到晶体结构的数据。
3．利用已建立的药理模型对所合成的化合物进行生物活性评价，并对构效关系做了定性分析。鉴定PPAR激动剂生物活性的方法主要分为两部分，即分别测定分子的结合活性和功能活性。对所合成的53个化合物测定了化合物与PPARγ的结合能力；对所合成的82个化合物在10~(-5)M测定了转染PPARγ基因的细胞水平上的转录激活实验；然后从中选取活性较好的24个化合物，在六个浓度测定了转染PPARα，PPARγ基因的细胞水平上的转录激活实验。实验结果显示：α-烷氧丙酸类化合物FJ-08-46的活性较好，可以此化合物为先导物进行结构改造，期望获得高效、低毒的2型糖尿病的治疗药。目前化合物FJ-08-46体内药效学评价正在进行中。
4．PPARγ激动剂有抑制肿瘤生长和预防治疗癌症的作用，对所合成目标化合物进行了体外MTT法和集落形成法抑制肿瘤细胞生长实验和体内肿瘤模型裸鼠的动物实验。实验结果显示化合物FJ-05-05对结肠癌HT-29模型裸鼠肿瘤有一定抑制作用，可根据此模型进行筛选，期望找到高效低毒的抗癌药物。
Type 2 diabetes is a metabolic disease characterized by insulin resistance, hyperglycaemia, and hyperlipidemia, leading to impaired secretion of insulin in the later stsges. The disease is often associated with obesity, dyslipidemia, and hypertension , causing increased cardiovascular risks. Discovery of the insulin-sensitizing drugs thiazolidinedione(TZD) and the drug-target (Peroxisome proliferator-activated receptors, PPARs) offers a new route for the treatment of type 2 diabetes. PPARs are members of a huge nuclear hormone receptor superfamily, a group of nuclear proteins that mediate the specific effects of small lipophhilic compounds, such as steroids, retinoids, and fatty acids, on DNA transcription. Three subtypes of PPARs have been identified and cloned from mouse and human: PPARα, PPARγ, and PPARδ. To date three PPARγagonists Troglitazone, Rosiglitazone and Pioglitazone have been launched, which, however, provide minor protection against eventual cardiovascular risks that develop with type 2 diabetes. A new strategy of designing novel anti-diabetics is to combine the insulin sensitizing and glucose regulating activity of a PPARγagonists with lipid/cholesterol modulating properties of PPARαagonists (fibrate) into a single molecule. Dual-acting PPARαand PPARγagonists, designed to combine the beneficial effects seen with insulin sensitizers and fibrate, could be new drugs to 2 type diabetes.In order to find new and more effective drugs for the type 2 diatebes, the following work had been carried out in the thesis:1. Scrutinizing the structures of PPAR agonists and the binding feature of agonists to the receptors, a novel virtual library of PPAR agonists was constructed by structure-based drug design and combinatorial library method. Virtual screening was conducted by DOCKoperation, which automatically simulated the interaction between the ligands and receptors. Those strong-binding virtual compounds were selected according to the scoring functions. Six series of compounds composed of diverse building blocks were designed and synthesized in order to search for lead compounds with dual activity.
X=H, N
R_1=H, -C_2H_5
R2=-CH_2OH, -CH_2COOH
R3=-CH_3, -C_2H_5, -iC_3H_7, -C_5H_9, -C_6H_(11), -C_6H_(5…)
2. One hundred and forty-five chemical substances including ninety new compounds were prepared, of which eighty-five novel compounds are target molecules. All the targets moleculars were identified by ~1HNMR (300MHz) and MS (EI, FAB, or ESI), twenty-four of which were identified by high revolution mass spectroscopy. The compound FJ-07-17 was also identified by X-ray crystallography.
3. Most of the target compounds were evaluated in vitro. Some of the compounds evaluated show strong binding affinities to PPARγ. Some of the compounds were found possessing approximate activity to Pioglitazone in vitro PPARαand PPARγtransactivation. The Structure-activity relationships of synthsized compounds and transactivation data offered detailed molecular information for design and synthesis of new compounds. The evaluation of compound FJ-08-46 in vivo is in progresss.
4. Some compounds were in vitro tested for the activity on cell differentiation and proliferation. The compound FJ-05-05 was found to exhibit moderate inhibition of HT-29 cell line.
引文
1. Cobb, J. E., Blanchard, S. G. Boswell, E John M. N. and Allan S. W., ANNUAL REPORTS IN MEDICINAL CHEMISTRY, 2000, Vol. 35, 211-218.
2. Collins, J. L., Blanchard, S. G., Evan, B. G., J. Med. Chem., 1998, 41, 5037.
3. Issemann, I., Green, S., Nature, 1990, 347, 645-650.
4. David, J., Mangelsdorf, Carl Thummel, Miguel Beato, Cell, 1995, Vol.83,835-839.
5. Joseph Vamecq, Norbert Latruffe, Lancet, 1999.
6. Iwata M., Haruta T., Usui I., et al., Diabetes, 2001, 50(5), 1083-1092.
7. Young P. W., Cawthorne M.A., Coyle P.J., et al., Diabetes 1995, 44(9), 1087-1092.
8. Hofmann C., Lorenz K., Braithwaite S. S., et al., Endocrinology, 1994, 134(1), 264-270.
9. Kallen C. B., Lazar M. A., Proc. Natl. Acad. Sci. USA, 1996, 93(12), 5793-5796.
10. Shao D et al, J Biol Chem, 1997, 272, 21473.
11. Smith M. T., Chemical Research in Toxicology, 2003, 16(6), 679-687.
12. Ekins S., Schuetz E., Trends Pharmacol. Sci., 2002, 23(2), 49-50.
13. Kubota T, Koshizuka K, Williamson EA, Cancer Res. 1998 Aug 1;58(15):3344-52.
14. Willson, T. M., Brown, P. J., Sternbach, D. D., J. Med. Chem., 2000, 43,527-550.
15. Barry G. S. and William J. H., Current Medicinal Chemistry, 2003, 10(4), 267-280.
16. Brown, P. J., Winegar, D. A., Plunket, K. D. et al., J. Med. Chem., 1999, 42, 3785-3788.
17. Kliewer S. A., Sundseth S. S., Jones S. A. et al., Proc. Natl. Acad. Sci. USA, 1997, 94(9), 4318-4323.
18. Forman B. M., Chen J., Evans R. M., Proc. Natl. Acad. Sci. USA, 1997, 94(9), 4312-4317.
19. Davies S. S., Pontsler A. V., Marathe G. K., J. Biol. Chem., 2001, 276(19), 16015-16023.
20. Cantello, B.C.C., Lawthorne, M.A., Cottam, G.P., J. Med. Chem., 1994, 37, 3977-3985.
21. McDonnell D. E, Vegeto, E., Gleeson, M. A.. Biotechnology, 1993, 11(11), 1256-1261.
22. Yasuhisa Kurogi, Drug Design and Discovery, 1999, Vol. 16, 109-118.
23. Robert, T. N., Stefan W., Nature, 1998, Vol.395, 137-143.
24. Dow, R. L., Beche, B. M., Chou, T. Y., J. Med. Chem., 1991, 34, 1538-1544.
25. Goldstein, S. W., McDermott, R. E., Gibbs, E. M.et al., J. Med. Chem.. 1993, 36. 2238-2240.
26. Shinkai, H., Onogi, S., Tanaka, M., et al., J. Med. Chem., 1998, 41, 1927-1933.
27. Parks, D. J., Tomkinson, N. C. O., Villeneuve, M. S., Bioorg. Med. Chem. Lett., 1998, 8, 3657-3658.
28. Buckle, D. R., Cantello, B. C. C., Cawthorne, M. A. et al, Bioorg. Med. Chem. Lett., 1996, 6, 2121-2126.
29. Henke, B. R., Blanchard, S. G., Brackeen, M. F., J. Med. Chem., 1998, 41, 5020.
30. Collins, J.L., Blanchard, S.G., Evan, B.G., J. Med. Chem., 1998, 41, 5037.
31. Cobb, J.E., Blanchard, S.G. Boswell, E.G., J. Med. Chem., 1998, 41, 5055.
32. Gampe R. T., Montana V. G., Lambert M. H., Mol. Cell., 2000, 5,545-555.
33. Elbrecht A., Chen Y., Adams A. et al., J. Biol. Chem., 1999, 274(12), 7913-7922.
34. Oberfield J. L., Collins J. L., Holmes C. P. et al., Proc. Natl. Acad. Sci. USA, 1999, 96(11), 6102-6106.
35. Pickavane L., Widdowson P. S., King P. et al., Br. J. Pharmacol., 1998, 125(4), 767-770.
36. Huang, J. T., Welch, J. S., Ricote, M. et al., Nature, 1999, 400, 378-382.
37. Oliver, W. R., Shenk, J. L., Snaith, M. R. et al., Proc. Natl. Acad. Sci. USA, 2001, 98, 5306-5311.
38. Berger, J., Leibowitz, M. D., Doebberr, T. W. et al., J. Biol. Chem., 1999, 274, 6718-6725.
39．蔡哲峰，郭宗儒，药学学报．2004，39(2)，158-160．
40. Lohray B. B., Bhushan V.. Rao B. P. et al., J. Med. Chem., 1994, 41(10), 1619-1630.
41. Brooks, D. A., Etgen, G. J., Rito, C. J. et al., J. Med. Chem., 2001,44, 2061-2064.
42. Lohray, B. B., Lohray, V. B., Bajji, A. C. et al., J. Med. Chem., 2001, 44, 2675.
43. Oakes, N. Abstract 446-P.62nd Sci. Sess. Am. Diabetes Assoc., June 14-18, 2002.
44. Drug Data Rep 2002, 24(5), 436.
45. Sorbera, L.A., Castaner, J., del Fresno, M. et al., Drugs Fut., 2002, 27(2), 132. Drug Data Rep 2001, 23(8), 788.
46. Drug Data Rep, 2003, 25(3), 247.
47．郭宗儒，药物化学总论(第二版)，2003
48. Cronet, R, Petersen, J. F. W., Folmer, R., et al., Structure, 2001, 9, 699-706.
49. Xu, H. E., Lambert, M. H., Montana, V. G. et al., Proc. Natl. Acad. Sci. USA, 2001, 98, 13919.
50. The program sybyl 6.8 is available from Tripos Assoc., 1699 S. Hanley Rd., St. Louis, MO 63144. 1998.
51. Ewing, T. J. A., Makino, S., Skillman, A. G., Kuntz, I. D. Journal of Computer-Aided Molecular Design, 2001, 15, 411.
52. Saito, N., Tashiro, K., Maru, Y. J Chem Soc, Perkin Trans, 1 1997, (1), 53-69.
53. Kende A. S., Bentley T. J., Mader R. A., J. Amer. Chem. Soc., 1974, 96, 4332.
54. Gurjar M. K., Bagavant G. Indian J. Chem., 1976, Vol. 14B, 548-549.
55. Giacomelli, G., Porcheddu, A., Salaris, M., Org.Lett., 2003, 5(15), 2715-2718.
56. Willy L. Tetrahedran Letters. 1970, 54, 4723-4724.
57. Srinivasascchari R., Sharada J. S. Tetrahedron, 1986, 42(20), 5739-5746.
58．王敏敏论文．
59．俞凌翀，有机化学中的人名反应，1984，305
60. Xingjun Fang, Jinmao You, Tianquan Jiao, Org. Prep. Proced. Int. 2000, 32(3), 284-287.
61. Loughlin W.A., McCleary M. A., HealyAust. P. C., Aust. J. Chem., 1984, 37, 1791-1794.
62. Robert O. H., David R., Nicholas N. et al., J. Org. Chem., 41(20), 1976, 3328-3329.
63．成峰，沈建华，罗小民等，中国药物化学杂志，2003，13(2)，110-118．
64. Stephane R., Frederic P., Joseph V., Molecular Cell, Vol. 8, 737-747, October, 2001,
2. Collins, J. L., Blanchard, S. G., Evan, B. G., J. Med. Chem., 1998, 41, 5037.
3. Issemann, I., Green, S., Nature, 1990, 347, 645-650.
4. David, J., Mangelsdorf, Carl Thummel, Miguel Beato, Cell, 1995, Vol.83,835-839.
5. Joseph Vamecq, Norbert Latruffe, Lancet, 1999.
6. Iwata M., Haruta T., Usui I., et al., Diabetes, 2001, 50(5), 1083-1092.
7. Young P. W., Cawthorne M.A., Coyle P.J., et al., Diabetes 1995, 44(9), 1087-1092.
8. Hofmann C., Lorenz K., Braithwaite S. S., et al., Endocrinology, 1994, 134(1), 264-270.
9. Kallen C. B., Lazar M. A., Proc. Natl. Acad. Sci. USA, 1996, 93(12), 5793-5796.
10. Shao D et al, J Biol Chem, 1997, 272, 21473.
11. Smith M. T., Chemical Research in Toxicology, 2003, 16(6), 679-687.
12. Ekins S., Schuetz E., Trends Pharmacol. Sci., 2002, 23(2), 49-50.
13. Kubota T, Koshizuka K, Williamson EA, Cancer Res. 1998 Aug 1;58(15):3344-52.
14. Willson, T. M., Brown, P. J., Sternbach, D. D., J. Med. Chem., 2000, 43,527-550.
15. Barry G. S. and William J. H., Current Medicinal Chemistry, 2003, 10(4), 267-280.
16. Brown, P. J., Winegar, D. A., Plunket, K. D. et al., J. Med. Chem., 1999, 42, 3785-3788.
17. Kliewer S. A., Sundseth S. S., Jones S. A. et al., Proc. Natl. Acad. Sci. USA, 1997, 94(9), 4318-4323.
18. Forman B. M., Chen J., Evans R. M., Proc. Natl. Acad. Sci. USA, 1997, 94(9), 4312-4317.
19. Davies S. S., Pontsler A. V., Marathe G. K., J. Biol. Chem., 2001, 276(19), 16015-16023.
20. Cantello, B.C.C., Lawthorne, M.A., Cottam, G.P., J. Med. Chem., 1994, 37, 3977-3985.
21. McDonnell D. E, Vegeto, E., Gleeson, M. A.. Biotechnology, 1993, 11(11), 1256-1261.
22. Yasuhisa Kurogi, Drug Design and Discovery, 1999, Vol. 16, 109-118.
23. Robert, T. N., Stefan W., Nature, 1998, Vol.395, 137-143.
24. Dow, R. L., Beche, B. M., Chou, T. Y., J. Med. Chem., 1991, 34, 1538-1544.
25. Goldstein, S. W., McDermott, R. E., Gibbs, E. M.et al., J. Med. Chem.. 1993, 36. 2238-2240.
26. Shinkai, H., Onogi, S., Tanaka, M., et al., J. Med. Chem., 1998, 41, 1927-1933.
27. Parks, D. J., Tomkinson, N. C. O., Villeneuve, M. S., Bioorg. Med. Chem. Lett., 1998, 8, 3657-3658.
28. Buckle, D. R., Cantello, B. C. C., Cawthorne, M. A. et al, Bioorg. Med. Chem. Lett., 1996, 6, 2121-2126.
29. Henke, B. R., Blanchard, S. G., Brackeen, M. F., J. Med. Chem., 1998, 41, 5020.
30. Collins, J.L., Blanchard, S.G., Evan, B.G., J. Med. Chem., 1998, 41, 5037.
31. Cobb, J.E., Blanchard, S.G. Boswell, E.G., J. Med. Chem., 1998, 41, 5055.
32. Gampe R. T., Montana V. G., Lambert M. H., Mol. Cell., 2000, 5,545-555.
33. Elbrecht A., Chen Y., Adams A. et al., J. Biol. Chem., 1999, 274(12), 7913-7922.
34. Oberfield J. L., Collins J. L., Holmes C. P. et al., Proc. Natl. Acad. Sci. USA, 1999, 96(11), 6102-6106.
35. Pickavane L., Widdowson P. S., King P. et al., Br. J. Pharmacol., 1998, 125(4), 767-770.
36. Huang, J. T., Welch, J. S., Ricote, M. et al., Nature, 1999, 400, 378-382.
37. Oliver, W. R., Shenk, J. L., Snaith, M. R. et al., Proc. Natl. Acad. Sci. USA, 2001, 98, 5306-5311.
38. Berger, J., Leibowitz, M. D., Doebberr, T. W. et al., J. Biol. Chem., 1999, 274, 6718-6725.
39．蔡哲峰，郭宗儒，药学学报．2004，39(2)，158-160．
40. Lohray B. B., Bhushan V.. Rao B. P. et al., J. Med. Chem., 1994, 41(10), 1619-1630.
41. Brooks, D. A., Etgen, G. J., Rito, C. J. et al., J. Med. Chem., 2001,44, 2061-2064.
42. Lohray, B. B., Lohray, V. B., Bajji, A. C. et al., J. Med. Chem., 2001, 44, 2675.
43. Oakes, N. Abstract 446-P.62nd Sci. Sess. Am. Diabetes Assoc., June 14-18, 2002.
44. Drug Data Rep 2002, 24(5), 436.
45. Sorbera, L.A., Castaner, J., del Fresno, M. et al., Drugs Fut., 2002, 27(2), 132. Drug Data Rep 2001, 23(8), 788.
46. Drug Data Rep, 2003, 25(3), 247.
47．郭宗儒，药物化学总论(第二版)，2003
48. Cronet, R, Petersen, J. F. W., Folmer, R., et al., Structure, 2001, 9, 699-706.
49. Xu, H. E., Lambert, M. H., Montana, V. G. et al., Proc. Natl. Acad. Sci. USA, 2001, 98, 13919.
50. The program sybyl 6.8 is available from Tripos Assoc., 1699 S. Hanley Rd., St. Louis, MO 63144. 1998.
51. Ewing, T. J. A., Makino, S., Skillman, A. G., Kuntz, I. D. Journal of Computer-Aided Molecular Design, 2001, 15, 411.
52. Saito, N., Tashiro, K., Maru, Y. J Chem Soc, Perkin Trans, 1 1997, (1), 53-69.
53. Kende A. S., Bentley T. J., Mader R. A., J. Amer. Chem. Soc., 1974, 96, 4332.
54. Gurjar M. K., Bagavant G. Indian J. Chem., 1976, Vol. 14B, 548-549.
55. Giacomelli, G., Porcheddu, A., Salaris, M., Org.Lett., 2003, 5(15), 2715-2718.
56. Willy L. Tetrahedran Letters. 1970, 54, 4723-4724.
57. Srinivasascchari R., Sharada J. S. Tetrahedron, 1986, 42(20), 5739-5746.
58．王敏敏论文．
59．俞凌翀，有机化学中的人名反应，1984，305
60. Xingjun Fang, Jinmao You, Tianquan Jiao, Org. Prep. Proced. Int. 2000, 32(3), 284-287.
61. Loughlin W.A., McCleary M. A., HealyAust. P. C., Aust. J. Chem., 1984, 37, 1791-1794.
62. Robert O. H., David R., Nicholas N. et al., J. Org. Chem., 41(20), 1976, 3328-3329.
63．成峰，沈建华，罗小民等，中国药物化学杂志，2003，13(2)，110-118．
64. Stephane R., Frederic P., Joseph V., Molecular Cell, Vol. 8, 737-747, October, 2001,