抗血栓药物—糖蛋白Ⅱb/Ⅲa拮抗剂的合成与活性
摘要
血栓形成是心血管疾病的病因之一，在形成血栓的各种因素中血小板聚集起重要作用，其中纤维蛋白原结合糖蛋白Ⅱb／Ⅲa(GPⅡb／Ⅲa)受体是血小板凝聚最后的共同的一步，因此阻断纤维蛋白原(fibrinogen)和GPⅡb／Ⅲa的结合可以抑制各种激动剂(如胶原，凝血酶等)介导的血小板聚集，所以GPⅡb／Ⅲa拮抗剂有巨大的市场价值。
本论文共合成药效团为脒苯基和羧基的两类化合物：
(1)线形化合物：药效团之间以线性骨架支撑的化合物；(2)苯二氮(艹卓)化合物：药效团之间以苯二氮(艹卓)为支撑骨架的化合物。
(1)线形化合物
在设计和合成已有GPⅡb／Ⅲa拮抗剂的me-too化合物之前，用比较分子力场分析法(CoMFA)对文献报道的16个化合物建立了三维定量构效关系模型，提示该类化合物的立体场，静电场的分布对活性的影响。
依据所得的3D-QSAR模型和Me-too药物分子设计原理，以处于三期临床研究的Sibrafiban和Lamifiban为模型分子，通过改变氨基酸侧链，设计并合成具有知识产权的新化合物，其中GPI-3，GPI-4，GPI-8，GPI-9，GPI-12，GPI-13，GPI-15，GPI-23，GPI-26等具有和模型化合物相似的活性，GPI-3，GPI-4，GPI-22，GPI-23进行了体内活性评价，结果表明测试化合物在体内可以保持相当的稳定性，在较长的时间内对血小板有较高程度的抑制，优于模型分子，进一步的实验正在进行中。
(2)苯二氮(艹卓)化合物
苯二氮(艹卓)化合物的设计以SB公司开发的Lotrafiban为先导物，合成化合物的药理评价正在进行中。
本论文共合成120个化合物，其中新化合物71个，目标化合物36个。
The thrombus formation is one of the causes of the cardiovascular diseases, in which process platelet aggregation involves the central event mediated by the binding of the plasma protein fibrinogen to the glycoprotein IIb/IIIa receptor (GPIIb/IIIa) on the surface of activated platelet. This binding is final common pathway for platelet aggregation. Therefore GPIIb/IIIa antagonists can completely inhibit platelet aggregation regardless of the various agonist(collagen, thrombin et al) involved. So GPIIb/IIIa receptor antagonists have a huge market value.Two series of compounds with amidinophenyl and carbonyl groups were synthesized in this dissertation:(1)linear compounds: the two pharmacophores being separated by a linear skeleton; (2)benzodiazepine compounds: the pharmacophores being attached on the benzodiazepine scaffold. (1)linear compoundsBefore design and synthesis of me-too compounds of existing GPIIb/IIIa antagonists, a 3D-QSAR analysis was conducted with 16 reported compounds using CoMFA. The result indicated both steric and electrostatic fields affect the inhibitory activity.Based on 3-D QSAR model and me-too drug design principle, the new compounds with proprietary were designed and synthesized through the change of the side chains of amino acids of Sibrafiban and Lamifiban which were investigated in clinical phase III and used as model compounds. Among the synthesized target compounds, GPI-3, GPI-4, GPI-8, GPI-9, GPI-12, GPI-13, GPI-15, GPI-22 and GPI-23 showed the activities as potent as the model compounds. And GPI-3, GPI-4, GPI-12, GPI-13 were further assayed in vivo. The result demonstrated these compounds maintained the metabolic stability and inhibited platelet aggregation in high level with a prolonged duration of activity, superior to the model compounds. The further experiments were being carried on.
(2) benzodiazepine compounds
The benzodiazepine compounds were designed and synthesized by using Lotrafiban as lead compound. The biological assays are in test.
In this dissertation, 120 compounds were synthesized, in which 71 compounds were novel and 36 compounds is the target molecules and identified.
本论文共合成药效团为脒苯基和羧基的两类化合物：
(1)线形化合物：药效团之间以线性骨架支撑的化合物；(2)苯二氮(艹卓)化合物：药效团之间以苯二氮(艹卓)为支撑骨架的化合物。
(1)线形化合物
在设计和合成已有GPⅡb／Ⅲa拮抗剂的me-too化合物之前，用比较分子力场分析法(CoMFA)对文献报道的16个化合物建立了三维定量构效关系模型，提示该类化合物的立体场，静电场的分布对活性的影响。
依据所得的3D-QSAR模型和Me-too药物分子设计原理，以处于三期临床研究的Sibrafiban和Lamifiban为模型分子，通过改变氨基酸侧链，设计并合成具有知识产权的新化合物，其中GPI-3，GPI-4，GPI-8，GPI-9，GPI-12，GPI-13，GPI-15，GPI-23，GPI-26等具有和模型化合物相似的活性，GPI-3，GPI-4，GPI-22，GPI-23进行了体内活性评价，结果表明测试化合物在体内可以保持相当的稳定性，在较长的时间内对血小板有较高程度的抑制，优于模型分子，进一步的实验正在进行中。
(2)苯二氮(艹卓)化合物
苯二氮(艹卓)化合物的设计以SB公司开发的Lotrafiban为先导物，合成化合物的药理评价正在进行中。
本论文共合成120个化合物，其中新化合物71个，目标化合物36个。
The thrombus formation is one of the causes of the cardiovascular diseases, in which process platelet aggregation involves the central event mediated by the binding of the plasma protein fibrinogen to the glycoprotein IIb/IIIa receptor (GPIIb/IIIa) on the surface of activated platelet. This binding is final common pathway for platelet aggregation. Therefore GPIIb/IIIa antagonists can completely inhibit platelet aggregation regardless of the various agonist(collagen, thrombin et al) involved. So GPIIb/IIIa receptor antagonists have a huge market value.Two series of compounds with amidinophenyl and carbonyl groups were synthesized in this dissertation:(1)linear compounds: the two pharmacophores being separated by a linear skeleton; (2)benzodiazepine compounds: the pharmacophores being attached on the benzodiazepine scaffold. (1)linear compoundsBefore design and synthesis of me-too compounds of existing GPIIb/IIIa antagonists, a 3D-QSAR analysis was conducted with 16 reported compounds using CoMFA. The result indicated both steric and electrostatic fields affect the inhibitory activity.Based on 3-D QSAR model and me-too drug design principle, the new compounds with proprietary were designed and synthesized through the change of the side chains of amino acids of Sibrafiban and Lamifiban which were investigated in clinical phase III and used as model compounds. Among the synthesized target compounds, GPI-3, GPI-4, GPI-8, GPI-9, GPI-12, GPI-13, GPI-15, GPI-22 and GPI-23 showed the activities as potent as the model compounds. And GPI-3, GPI-4, GPI-12, GPI-13 were further assayed in vivo. The result demonstrated these compounds maintained the metabolic stability and inhibited platelet aggregation in high level with a prolonged duration of activity, superior to the model compounds. The further experiments were being carried on.
(2) benzodiazepine compounds
The benzodiazepine compounds were designed and synthesized by using Lotrafiban as lead compound. The biological assays are in test.
In this dissertation, 120 compounds were synthesized, in which 71 compounds were novel and 36 compounds is the target molecules and identified.
引文
[1](a) American Heart Association. 1997 Heart and strok statistical update.Dallas(TX): American Heart Assiciation.1997.
(b) Murray CJL, Lopez AD. Mortality by cause for eight regions of the world:global burden of disease study.Lancet 1997; 349:1269-76.
[2] American Heart Association. 1998 Heart and strok statistical update.Dallas(TX): American Heart Assiciation.1998.
[3] Davies MJ, Thomas AC. Plaque fissuring-the cause of acute myocardial infarction, sudden ischemic death, and crescendo angina. Br Heart J 1985; 53: 363-73 .
[4] Becker RC. Seminars in thrombosis, thrombolysis and vascular biology. 4.Fibrinolysis. Cardiology. 1991; 79: 188.
[5](a) Philips DR; Charo IF; Parise LV. Blood. 1988, 71:831
(b) Philips DR,Kieffer N, Annu. Rev. Cell Biol.1990, 6: 329.
[6] Coller BS. Platelets and Thrombolytic Therapy. N. Engl .J. Med. 1990; 322:33-42.
[7] Coller BS. Blockade of platelet GPIIb/IIIa receptors as an antithrombotic strategy. Circulation 1995; 92: 2373-80.
[8] Coller BS, Peerschke EI, Scudder LE,et al. A murine monoclonal antibody that completely blocks the binding of fibrinogen and binds to glycoprotein IIb and /or IIIa. J Clin Invest 1983; 72: 325-38.
[9] Cook NS, Ubben D. Fibrinogen as a major risk factor in cardiovascular disease. Trends Pharmacol Sci. 1991;11: 444-51.
[10] Marguerie GA, Ardaillou N, Cherel G, et al. The binding of fibrinogen to its platelet receptor. J Biol Chem 1982, 257:11872-5.
[11] Pytela R, Pierschbacher MD, geinsberg MH, et al. Platelet membrane glycoprotein IIb/IIIa: Member of a family of Arg-Gly-Asp-specific adhesion molecules. Science 1986, 231: 1559-62.
[12] Cook NS, Zerwes HG, Tapparelli C, et al. Platelet aggregation and fibrinogen binding in human, rhesus monkey, guinea-pig, hamster and rat blood:activation by ADP and a thrombin receptor peptide and inhibition by glycoprotein IIb/IIIa antagonist.Thromb Haemostas. 1993, 70: 531-9.
[13]Pierschbacher MD, Ruoslahti E. Influence of Stereochemistry of the Sequence Arg-Gly-Asp-Xaa on Binding Specificity in Cell Adheion. J. Biol Chem. 1987,262: 17294-17298。
[14] Lefkovits J, Plow EF, Topol EJ. Platelet glycoprotein IIb/IIIa receptor in cardiovascular medine. N Engl J Med, 1995: 332(23): 1553-1559.
[15] Huang TF, Holt JC, Lukasiewicz H, et al. A low molecular weight peptide inhibiting fibrinogen interaction with platelet receptors expressed on glycoprotein IIb/IIIa complex. J Biol Chem. 1987, 262: 16153-63.
[16] Mcdowell RS, Gadek TR. Structural studies of potent constrained RGD peptide. J Amer Chem Soc. 1992, 114(24): 9245-53.
[17] Gold HK, Gimple LW, Leinbach rc et al. Pharmacodynamic Study of F(ab')_2 Fragment of Murine Monoclonal Antibody with Unstable Angina Pectoris. J.Clin. Invest. 1990, 86: 651-659.
[18] Zablocki JA, Miyano M, Garland RB, et al. Potent in vitro and in vivo inhinbitors of platelet aggregation based upon the Arg-Gly-Asp-Phe sequence of fibrinogen. A proposal on the nature of the binding interaction between the Arg-guanidine of RGDX mimetics and the platelet GPIIb/IIIa receptor. J Med Chem, 1993,36:1811.
[19](a) Nicholson N, Taite B, Panzer-Knodle S, et al. An orally active glycoprotein IIb/IIIa antagonist SC 54684. Thromb Haemostas, 1993, 69: 975
(b)Zablocki J, Nicholson N, Taite B, et al. Selection an orally active glycoprotein
IIb/IIIa receptor antagonist for clinical trials. Thromb, Haemostas 1993, 69:1244.
[20] Alig L, Endenhofer A, Hadvary P, et al. Low molecular weight non-peptide fibrinogen receptor antagonists. J Med Chem, 1992, 35: 4393-407.
[21] Weller J, Alig L, Bersini M, et al. Orally active fibrinogen receptor antagonist .2. amidoximes as amidines, J Med Chem, 1996, 39: 3139-3147.
[22] Hartman GD, Egbertson MS, Halczenko W, et al. Non-peptide fibrinogen receptor antagonists, 1. discovery and design of exosite inhinbtors. J Med Chem,1992,35:4640-2.
[23] Egbertson MS, Hartman GD, Gould RJ, et al. Non-peptide GPIIb/IIIa inhibitors. 10. Centrally constrained alpha-sulfonamides are potent inhibitors of platelet aggregation. Bioorg Med Chem lett, 6(21): 2519-2514.
[24] Halczenko W, Cook JJ, Holahan MA. Nonpeptide glycoprotein IIb/IIIa inhibitors. 12. Potent and orally active centrally constrained thieno[2,3-c]pyridones, Bioorg Med Chem Lett, 1996, 6(22): 2771-2776.
[25] Brashear KM, Cook JJ, Bendnar B, et al. Nonpeptide glycoprotein Ilb/IIIa inhibitors. 18. Indole alpha-sulfonamide acids are potent inhibitorsof platelet aggregation, Bioorg Med Chem Lett, 1997, 7(21), 2793-2798.
[26] Peishoff CE, Ali FE, Bean JW et al. Investigation of conformational specificity at GPIIb/IIIa: Evaluation of constrained RGD peptides. J Med Chem,1992,35:3962-9.
[27] Bondinell WE, Callahan JF, Huffman WF, et al. Bicyclic fibrinogen receptor antagonists, WO 9300095, 1993.
[28]Blackburn B, Barker P, Gadek T, et al. Non-peptidyl intergrin inhibitors having specificity for the GPIIb/IIIa receptor. WO9308174, 1993.
[29] Himmelsbach F, Austel V, Pieper H, et al. EP528369-A, EP483667-A,1993.
[30] Porter B, Eldred CD, Kelly HA, et al. Piperidineacetic acid derivativees as inhibitors of fibrinogen-dependent blood platelet aggregation. EP 542363-A, WO 9310091, EP 537980-A, WO9314077, 1993
[31] Alig L, Hadvary P, Hiirzeler M, et al. United States Patent, 5,378,712.
[32] Wright SW, Hageman DL, Wright AS, et al. Convenient preparation of t-Butyl esters and ethers from t-Butanol, Tetrahedron Letter, 389420; 7345-48.
[33] 黄惟德 陈常庆．多肽合成，科学出版社，1995：13-44．
[34] 黄惟德 陈常庆．多肽合成，科学出版社，1995：102-140．
[35] X-衍射分析结果由中国医学科学院药物研究所吕扬，赵斌，郑启泰提供．
[36] Bergmann ED, Berkovic S, Ikan R. A new method for the preparation of Aromatic fluorine compounds. J Am Chem Soc, 1956, 6037-6039
[37] 韩广甸，赵树纬 李述文等．有机制备化学手册，中卷， 31-32．
[38] Klein SI, Molino BF, Czekaj M, et al. Design of a new class of orally active fibrinogenreceptor antagonists. J Med Chem, 1998, 41: 2492-2502.
[39] CN 88100802-A.
[40] Organic syntheses volume 5,373-375.
[41] 黄惟德 陈常庆．多肽合成，ppll．
[42] 黄惟德 陈常庆．多肽合成，pp91．
[43] Dvies JS, Mohammed AKA, Chiral analysis of the reaction stages in the edman method for sequencing peptides. J Chem Soc. Perkin Trans Ⅱ,1984,1723-1727.
[44] Baltzly R, Sheehan LW, Stone A. J Org Chem, 1961, 26(7): 2353.
[45] Chan RL, Bruice TCm. The chemistry of an electron-deficient 5-deazaflavin. 8-cyano-10-methyl-5-deazaisoalloxazine., 1977,99,(20):6721-30.
(b) Murray CJL, Lopez AD. Mortality by cause for eight regions of the world:global burden of disease study.Lancet 1997; 349:1269-76.
[2] American Heart Association. 1998 Heart and strok statistical update.Dallas(TX): American Heart Assiciation.1998.
[3] Davies MJ, Thomas AC. Plaque fissuring-the cause of acute myocardial infarction, sudden ischemic death, and crescendo angina. Br Heart J 1985; 53: 363-73 .
[4] Becker RC. Seminars in thrombosis, thrombolysis and vascular biology. 4.Fibrinolysis. Cardiology. 1991; 79: 188.
[5](a) Philips DR; Charo IF; Parise LV. Blood. 1988, 71:831
(b) Philips DR,Kieffer N, Annu. Rev. Cell Biol.1990, 6: 329.
[6] Coller BS. Platelets and Thrombolytic Therapy. N. Engl .J. Med. 1990; 322:33-42.
[7] Coller BS. Blockade of platelet GPIIb/IIIa receptors as an antithrombotic strategy. Circulation 1995; 92: 2373-80.
[8] Coller BS, Peerschke EI, Scudder LE,et al. A murine monoclonal antibody that completely blocks the binding of fibrinogen and binds to glycoprotein IIb and /or IIIa. J Clin Invest 1983; 72: 325-38.
[9] Cook NS, Ubben D. Fibrinogen as a major risk factor in cardiovascular disease. Trends Pharmacol Sci. 1991;11: 444-51.
[10] Marguerie GA, Ardaillou N, Cherel G, et al. The binding of fibrinogen to its platelet receptor. J Biol Chem 1982, 257:11872-5.
[11] Pytela R, Pierschbacher MD, geinsberg MH, et al. Platelet membrane glycoprotein IIb/IIIa: Member of a family of Arg-Gly-Asp-specific adhesion molecules. Science 1986, 231: 1559-62.
[12] Cook NS, Zerwes HG, Tapparelli C, et al. Platelet aggregation and fibrinogen binding in human, rhesus monkey, guinea-pig, hamster and rat blood:activation by ADP and a thrombin receptor peptide and inhibition by glycoprotein IIb/IIIa antagonist.Thromb Haemostas. 1993, 70: 531-9.
[13]Pierschbacher MD, Ruoslahti E. Influence of Stereochemistry of the Sequence Arg-Gly-Asp-Xaa on Binding Specificity in Cell Adheion. J. Biol Chem. 1987,262: 17294-17298。
[14] Lefkovits J, Plow EF, Topol EJ. Platelet glycoprotein IIb/IIIa receptor in cardiovascular medine. N Engl J Med, 1995: 332(23): 1553-1559.
[15] Huang TF, Holt JC, Lukasiewicz H, et al. A low molecular weight peptide inhibiting fibrinogen interaction with platelet receptors expressed on glycoprotein IIb/IIIa complex. J Biol Chem. 1987, 262: 16153-63.
[16] Mcdowell RS, Gadek TR. Structural studies of potent constrained RGD peptide. J Amer Chem Soc. 1992, 114(24): 9245-53.
[17] Gold HK, Gimple LW, Leinbach rc et al. Pharmacodynamic Study of F(ab')_2 Fragment of Murine Monoclonal Antibody with Unstable Angina Pectoris. J.Clin. Invest. 1990, 86: 651-659.
[18] Zablocki JA, Miyano M, Garland RB, et al. Potent in vitro and in vivo inhinbitors of platelet aggregation based upon the Arg-Gly-Asp-Phe sequence of fibrinogen. A proposal on the nature of the binding interaction between the Arg-guanidine of RGDX mimetics and the platelet GPIIb/IIIa receptor. J Med Chem, 1993,36:1811.
[19](a) Nicholson N, Taite B, Panzer-Knodle S, et al. An orally active glycoprotein IIb/IIIa antagonist SC 54684. Thromb Haemostas, 1993, 69: 975
(b)Zablocki J, Nicholson N, Taite B, et al. Selection an orally active glycoprotein
IIb/IIIa receptor antagonist for clinical trials. Thromb, Haemostas 1993, 69:1244.
[20] Alig L, Endenhofer A, Hadvary P, et al. Low molecular weight non-peptide fibrinogen receptor antagonists. J Med Chem, 1992, 35: 4393-407.
[21] Weller J, Alig L, Bersini M, et al. Orally active fibrinogen receptor antagonist .2. amidoximes as amidines, J Med Chem, 1996, 39: 3139-3147.
[22] Hartman GD, Egbertson MS, Halczenko W, et al. Non-peptide fibrinogen receptor antagonists, 1. discovery and design of exosite inhinbtors. J Med Chem,1992,35:4640-2.
[23] Egbertson MS, Hartman GD, Gould RJ, et al. Non-peptide GPIIb/IIIa inhibitors. 10. Centrally constrained alpha-sulfonamides are potent inhibitors of platelet aggregation. Bioorg Med Chem lett, 6(21): 2519-2514.
[24] Halczenko W, Cook JJ, Holahan MA. Nonpeptide glycoprotein IIb/IIIa inhibitors. 12. Potent and orally active centrally constrained thieno[2,3-c]pyridones, Bioorg Med Chem Lett, 1996, 6(22): 2771-2776.
[25] Brashear KM, Cook JJ, Bendnar B, et al. Nonpeptide glycoprotein Ilb/IIIa inhibitors. 18. Indole alpha-sulfonamide acids are potent inhibitorsof platelet aggregation, Bioorg Med Chem Lett, 1997, 7(21), 2793-2798.
[26] Peishoff CE, Ali FE, Bean JW et al. Investigation of conformational specificity at GPIIb/IIIa: Evaluation of constrained RGD peptides. J Med Chem,1992,35:3962-9.
[27] Bondinell WE, Callahan JF, Huffman WF, et al. Bicyclic fibrinogen receptor antagonists, WO 9300095, 1993.
[28]Blackburn B, Barker P, Gadek T, et al. Non-peptidyl intergrin inhibitors having specificity for the GPIIb/IIIa receptor. WO9308174, 1993.
[29] Himmelsbach F, Austel V, Pieper H, et al. EP528369-A, EP483667-A,1993.
[30] Porter B, Eldred CD, Kelly HA, et al. Piperidineacetic acid derivativees as inhibitors of fibrinogen-dependent blood platelet aggregation. EP 542363-A, WO 9310091, EP 537980-A, WO9314077, 1993
[31] Alig L, Hadvary P, Hiirzeler M, et al. United States Patent, 5,378,712.
[32] Wright SW, Hageman DL, Wright AS, et al. Convenient preparation of t-Butyl esters and ethers from t-Butanol, Tetrahedron Letter, 389420; 7345-48.
[33] 黄惟德 陈常庆．多肽合成，科学出版社，1995：13-44．
[34] 黄惟德 陈常庆．多肽合成，科学出版社，1995：102-140．
[35] X-衍射分析结果由中国医学科学院药物研究所吕扬，赵斌，郑启泰提供．
[36] Bergmann ED, Berkovic S, Ikan R. A new method for the preparation of Aromatic fluorine compounds. J Am Chem Soc, 1956, 6037-6039
[37] 韩广甸，赵树纬 李述文等．有机制备化学手册，中卷， 31-32．
[38] Klein SI, Molino BF, Czekaj M, et al. Design of a new class of orally active fibrinogenreceptor antagonists. J Med Chem, 1998, 41: 2492-2502.
[39] CN 88100802-A.
[40] Organic syntheses volume 5,373-375.
[41] 黄惟德 陈常庆．多肽合成，ppll．
[42] 黄惟德 陈常庆．多肽合成，pp91．
[43] Dvies JS, Mohammed AKA, Chiral analysis of the reaction stages in the edman method for sequencing peptides. J Chem Soc. Perkin Trans Ⅱ,1984,1723-1727.
[44] Baltzly R, Sheehan LW, Stone A. J Org Chem, 1961, 26(7): 2353.
[45] Chan RL, Bruice TCm. The chemistry of an electron-deficient 5-deazaflavin. 8-cyano-10-methyl-5-deazaisoalloxazine., 1977,99,(20):6721-30.