N-芳基取代异硫脲类H～+/K～+-ATP酶抑制剂的合成及活性研究
摘要
H~+／K~+-ATP酶抑制剂是继组胺H_2-受体拮抗剂之后一种新型的抑酸类抗溃疡药物。本文简述了H~+／K~+-ATP酶抑制剂的作用机制和发展概况。在前期工作的基础上，设计、合成了吡啶和喹啉取代的两类异硫脲化合物，研究其对H~+／K~+-ATP酶抑制作用的构效关系。本文共合成了20个未见文献报道的目标化合物，并经~1H-NMR，~(13)C-NMR和MS确证其结构。
采用大鼠游离胃灌注方法考察目标化合物对胃酸分泌的影响，结果表明：其中有12个目标化合物的抑酸活性强于或相当于阳性对照药泮妥拉唑钠，化合物BTU-08的抑酸作用最强，为阳性对照药的1.7倍。喹啉环取代的目标化合物的活性优于吡啶环取代的目标化合物。
根据测定的药理活性数据，应用CoMFA技术研究了该类化合物的QSAR，得到了电性效应及场效应对该类化合物活性的影响规律，在此规律基础上对能够增强活性的化合物结构给予预测，为进一步结构修饰以获得更好活性的H~+／K~+-ATP酶抑制剂提供了指导。
H+/K+-ATPase inhibitor has become novel type of antiulcer drug with inhibition of gastric acid secretion after the appearance of histamine H2-receptor antagonist. In this thesis we briefly summarized the mechanism and the progress of the H+/K+-ATPase inhibitors. On the base of our former research work, we designed and synthesized pyridine and quinoline derivatives as new types isothioureas to study the QSAR of the H+/K+-ATPase inhibitor. We totally synthesized 20 new N-arylisothiourea derivatives and indentified their structures by 1H-NMR, 13C-NMR and MS.
The method of affusing the dissociated stomach of the big rat was used to study the changing of the gastric acid secretion effected by the target compounds. The result shows that 12 of the target compounds have comparable or strong gastric acid inhibitory activities with the positive control sodium pantoprazolc. Especially, compound BTU-08 gave the most powerful activity which was nearly two times higher than the control. And the bioactivity of the quinoline derivatives is better than the pyridine derivatives.
Using the compound bioactivities we studied the QSAR of the N-arylisothioureas by CoMFA computation. And we got the regular how the steric factor and electrostatic factor effect on the bioactivity. And we also predicted the model structure which will have more powerful bioactivity based on the regular. This regular will benefit the work of modifying the molecular to find the better H+/K+-ATPase inhibitor in the future.
采用大鼠游离胃灌注方法考察目标化合物对胃酸分泌的影响，结果表明：其中有12个目标化合物的抑酸活性强于或相当于阳性对照药泮妥拉唑钠，化合物BTU-08的抑酸作用最强，为阳性对照药的1.7倍。喹啉环取代的目标化合物的活性优于吡啶环取代的目标化合物。
根据测定的药理活性数据，应用CoMFA技术研究了该类化合物的QSAR，得到了电性效应及场效应对该类化合物活性的影响规律，在此规律基础上对能够增强活性的化合物结构给予预测，为进一步结构修饰以获得更好活性的H~+／K~+-ATP酶抑制剂提供了指导。
H+/K+-ATPase inhibitor has become novel type of antiulcer drug with inhibition of gastric acid secretion after the appearance of histamine H2-receptor antagonist. In this thesis we briefly summarized the mechanism and the progress of the H+/K+-ATPase inhibitors. On the base of our former research work, we designed and synthesized pyridine and quinoline derivatives as new types isothioureas to study the QSAR of the H+/K+-ATPase inhibitor. We totally synthesized 20 new N-arylisothiourea derivatives and indentified their structures by 1H-NMR, 13C-NMR and MS.
The method of affusing the dissociated stomach of the big rat was used to study the changing of the gastric acid secretion effected by the target compounds. The result shows that 12 of the target compounds have comparable or strong gastric acid inhibitory activities with the positive control sodium pantoprazolc. Especially, compound BTU-08 gave the most powerful activity which was nearly two times higher than the control. And the bioactivity of the quinoline derivatives is better than the pyridine derivatives.
Using the compound bioactivities we studied the QSAR of the N-arylisothioureas by CoMFA computation. And we got the regular how the steric factor and electrostatic factor effect on the bioactivity. And we also predicted the model structure which will have more powerful bioactivity based on the regular. This regular will benefit the work of modifying the molecular to find the better H+/K+-ATPase inhibitor in the future.
引文
[1] 沈建民，抗消化性溃疡药物国内外研究进展．1995：1～49．
[2] W. L. Peterson and C. T. Richardson in M.H.Sleisenger and J.S. Fordantran, eds. Gastrointestinal, Diseas. Saunders Philadelphia, Vol.1, 1983, 708-724.
[3] 李瑜元，酸相关疾病治疗的抑酸水平及药物选择．中国消化杂志，2001，21(11) 645-646．
[4] A. H. Soll and J.I. Isenberg in M. H. Sleisengeer and J.S. Fordtan, eds. Gastro intestinal, Disease. Saunder Philadelphia, Vol. 1, 1983, 625-672.
[5] Burger's Medicinal Chemistry and Drug Discovery. Volume 2, Therapeutic Agents.
[6] M. M. Wolfe and A H. Soll. N Engl J Med, 1988, 319, 1707-1715.
[7] G Sachs, HH Chang, E Rabon, etc. A nonelectrogenic H pump in plasma membranes of dog stomach. J Biol Chem, 1976,251:7690～7689.
[8] J M Wolosin and J G Forte. Stimulation of oxyntic cells triggers H~-/K~--ATP ase membrabe. Am J Physiol, 1984, 246: C537-545.
[9] G. E shull and Sblingrel. Molecular cloning of the rat stomach H~-/K~--ATPase. J Biol them, 1986, 261:16788～16791.
[10] Laurence V, Jean-M R and Erick G. Fourier Transform Infrared Spectroscopy Study of the Secondary Structure of the Reconstitued Neurospora erasa Plasma Membrane H~--ATPase and of Its Membrane-associated Propteolytic Peptides. J Biol Chem, ]995, 270(30), 17685-17696.
[11] M Maeda, J Ishizaki and M Futai. Cdna cloning and sequence determination of pig gastric H~-/K~--ATPase. Biochim Biophys Res Commun, 1988, 156: 203～
209.
[12] M A Reuben, L S Lasater and G Sachs. Characterization of a β subunit of the gastric H~-/K~--ATPase, Proc Natl Acad Sci, USA 87:6767～6771.
[13] K Bamberg, F Mercier, ma Reuben, etc. Cdna cloning and membrane topology of the rabit gastric H~-/K~--ATPase α-subunit. Biochem Biophys Acta, 1992, 1131: 69-77.
[14] 翟建国，闫冬，程卯生．H~-／K~--ATP酶抑制剂研究进展．中国药物药物化学杂志，10(2)，146-151，2000．
[15] P Lindberg, P Nordberg and T alminger etc. The mechanism of action of the gastric acid secretion inhibitor omprazoie. Journal of Medicinal Chemistry, 29(8),1986
[16] N Sung, K Jeong, D Jeon, ect. Influence of N-substitued amino group on the insecticidal activity of 2-(n-octyl)-3-(n-propyl)isothiourea derivatives. Hah' guk Nong hwa Hakoehchi, 1995,38(2):163～167;(CA:123:191175b)
[17] S Devi, A Nayak and AS Mittra. Synthesisi of some substituted 5-thiazoliones for possible use as fungcides. J Indian Chem soc, 1984:335-337.
[18] 常志初，蒋勤和徐炳祥等S-(2-吡啶基-N-氧化物)-N-取代或N，N-双取代异硫脲氢溴酸盐的合成及抗菌活性．药学学报1995，30(4)：263-268．
[19] AL Salzman, AG Denenberg, I Ueta, etc. Induction and activity of oxide syntbase in cultured human intestinal epithelia] monolayer. Am J Phaysiol, 1996,270(4):G565～573
[20] D Jang, C Szaboand A C Murrel. S-Subsitued isothoureas are potent inhibitors of nitric oxide biosynthesis in cartilage. Eur J Pharmacol, 1996, 312(3): 341～347
[21] JF White, MCW Minchin and C Ennis. Antidepressant (arylalkyl)amines as GABA autoreceptor agonists. US 5422355(CA:123:prP169355g).
[22] 翟建国 N-芳基取代异硫脲类H~-／K~--ATP酶抑制剂的合成及活性研究．沈阳药科大学硕士论文，2000年6月
[23] 王德传，张亦华，彭司勋等，二硫戊环异硫脲类化合物的合成及其一氧化氮合酶抑制剂活性研究．中国药科大学学报，32(1)，1～7，2001．
[23] T. Koenig and J.S. Wieczorek. The reaction of trichloroacetyl chloride with 2-picoline N-oxide and pridylcarbinols. The Journl of Organic Chemistry, 33(4), 1531-1532,1968.
[24] 全国原料药工艺汇编．1005-1007，国家医药管理总局，1980年．
[25] W.Mathes and H.Schuely.Agew them Inter Ed Engl,1963 2,144
[26] 陈芬儿，有机药物合成法．北京：中国医药科学技术出版，1999年4月 (第一卷)，82-88
[27] 段行信，实用精细有机合成手册．北京：化学工业出版社，2000，1，366-368
[28] Masakazu Tamura, Yasuteru Urano and Kazuya Kikuchi etc. Superoxide Dismutase Activity of Iron(Ⅱ)TPEN Complex and its Oeriva tires. Chem Pharm Bull, 48(10) 1514-1518 (2000).
[29] Robert J. Ife, C a. Dyke, David J. Keeling ect. 2-[[4-Amino-2-pridine]methyl] sulful]benzimidazole H~-/K~--ATPase inhibitors. the relationship betten Pridine Basicity, Stability, and Acitivity. J Med Chem, 1989, 32,1970-1977
[30] V. Boekelheide and W. J. Linn. Rearrangements of N-Oxide. Anovel Synthesis of Pridyl Carbinol and Andehydes. J Amer Chem Soc, 76, 1286～1290 (1954)
[31] Thomas L. G, Christl. Heterocyclic Chemistry (third edition) 1997.
[32] R. Bodalski and A. R. Katritzky. New rearrangement types in reactions of acetic anhydride with2-alkylpridine 1-oxides. Tetrahedron letters, 1968,
257(3) 257-260.
[33] Shigeru Oae, Teijiro Kitao and Yoshinori Kitaoka. The Mechanism of the reaction of 2-piconline N-oxide with acetic anhydride. J Amer Chem Soc, 84, 3359(1962)
[34] 刘波，高惠强和周洵钧，合成芳基硫脲的新方法．化学通报 1994，(5)：42-43
[35] R. Jason Herr, J. louise Kuhler, Harold Mecker etc. A Convenient Method for the Preparation of Primary and Symmetrical N, N'-Disubstitued Thioreas. Synthesis 2000. No. 11.1569-1574
[36] C.R. Rasmussen, F. J. Villani, L. E. Weaner. B. Ereynolds etc. Improved Procedures for the Preparation of Cycloalkyl-,Arylalkyl-, and Arythioures. Synthesis, 1988, 6,456-459.
[37] 实用有机化学手册．李述文 范如霖编译 上海科技技术出版社，1986年，168-169
[38] Acros organics Catalogue of Fine Chemicals. International Edition, 2000-2001
[39] 计算机辅助设计---原理、方法及应用．陈凯先，蒋华良，嵇汝运，上海：上海科技技术出版社，2000：165～174
[2] W. L. Peterson and C. T. Richardson in M.H.Sleisenger and J.S. Fordantran, eds. Gastrointestinal, Diseas. Saunders Philadelphia, Vol.1, 1983, 708-724.
[3] 李瑜元，酸相关疾病治疗的抑酸水平及药物选择．中国消化杂志，2001，21(11) 645-646．
[4] A. H. Soll and J.I. Isenberg in M. H. Sleisengeer and J.S. Fordtan, eds. Gastro intestinal, Disease. Saunder Philadelphia, Vol. 1, 1983, 625-672.
[5] Burger's Medicinal Chemistry and Drug Discovery. Volume 2, Therapeutic Agents.
[6] M. M. Wolfe and A H. Soll. N Engl J Med, 1988, 319, 1707-1715.
[7] G Sachs, HH Chang, E Rabon, etc. A nonelectrogenic H pump in plasma membranes of dog stomach. J Biol Chem, 1976,251:7690～7689.
[8] J M Wolosin and J G Forte. Stimulation of oxyntic cells triggers H~-/K~--ATP ase membrabe. Am J Physiol, 1984, 246: C537-545.
[9] G. E shull and Sblingrel. Molecular cloning of the rat stomach H~-/K~--ATPase. J Biol them, 1986, 261:16788～16791.
[10] Laurence V, Jean-M R and Erick G. Fourier Transform Infrared Spectroscopy Study of the Secondary Structure of the Reconstitued Neurospora erasa Plasma Membrane H~--ATPase and of Its Membrane-associated Propteolytic Peptides. J Biol Chem, ]995, 270(30), 17685-17696.
[11] M Maeda, J Ishizaki and M Futai. Cdna cloning and sequence determination of pig gastric H~-/K~--ATPase. Biochim Biophys Res Commun, 1988, 156: 203～
209.
[12] M A Reuben, L S Lasater and G Sachs. Characterization of a β subunit of the gastric H~-/K~--ATPase, Proc Natl Acad Sci, USA 87:6767～6771.
[13] K Bamberg, F Mercier, ma Reuben, etc. Cdna cloning and membrane topology of the rabit gastric H~-/K~--ATPase α-subunit. Biochem Biophys Acta, 1992, 1131: 69-77.
[14] 翟建国，闫冬，程卯生．H~-／K~--ATP酶抑制剂研究进展．中国药物药物化学杂志，10(2)，146-151，2000．
[15] P Lindberg, P Nordberg and T alminger etc. The mechanism of action of the gastric acid secretion inhibitor omprazoie. Journal of Medicinal Chemistry, 29(8),1986
[16] N Sung, K Jeong, D Jeon, ect. Influence of N-substitued amino group on the insecticidal activity of 2-(n-octyl)-3-(n-propyl)isothiourea derivatives. Hah' guk Nong hwa Hakoehchi, 1995,38(2):163～167;(CA:123:191175b)
[17] S Devi, A Nayak and AS Mittra. Synthesisi of some substituted 5-thiazoliones for possible use as fungcides. J Indian Chem soc, 1984:335-337.
[18] 常志初，蒋勤和徐炳祥等S-(2-吡啶基-N-氧化物)-N-取代或N，N-双取代异硫脲氢溴酸盐的合成及抗菌活性．药学学报1995，30(4)：263-268．
[19] AL Salzman, AG Denenberg, I Ueta, etc. Induction and activity of oxide syntbase in cultured human intestinal epithelia] monolayer. Am J Phaysiol, 1996,270(4):G565～573
[20] D Jang, C Szaboand A C Murrel. S-Subsitued isothoureas are potent inhibitors of nitric oxide biosynthesis in cartilage. Eur J Pharmacol, 1996, 312(3): 341～347
[21] JF White, MCW Minchin and C Ennis. Antidepressant (arylalkyl)amines as GABA autoreceptor agonists. US 5422355(CA:123:prP169355g).
[22] 翟建国 N-芳基取代异硫脲类H~-／K~--ATP酶抑制剂的合成及活性研究．沈阳药科大学硕士论文，2000年6月
[23] 王德传，张亦华，彭司勋等，二硫戊环异硫脲类化合物的合成及其一氧化氮合酶抑制剂活性研究．中国药科大学学报，32(1)，1～7，2001．
[23] T. Koenig and J.S. Wieczorek. The reaction of trichloroacetyl chloride with 2-picoline N-oxide and pridylcarbinols. The Journl of Organic Chemistry, 33(4), 1531-1532,1968.
[24] 全国原料药工艺汇编．1005-1007，国家医药管理总局，1980年．
[25] W.Mathes and H.Schuely.Agew them Inter Ed Engl,1963 2,144
[26] 陈芬儿，有机药物合成法．北京：中国医药科学技术出版，1999年4月 (第一卷)，82-88
[27] 段行信，实用精细有机合成手册．北京：化学工业出版社，2000，1，366-368
[28] Masakazu Tamura, Yasuteru Urano and Kazuya Kikuchi etc. Superoxide Dismutase Activity of Iron(Ⅱ)TPEN Complex and its Oeriva tires. Chem Pharm Bull, 48(10) 1514-1518 (2000).
[29] Robert J. Ife, C a. Dyke, David J. Keeling ect. 2-[[4-Amino-2-pridine]methyl] sulful]benzimidazole H~-/K~--ATPase inhibitors. the relationship betten Pridine Basicity, Stability, and Acitivity. J Med Chem, 1989, 32,1970-1977
[30] V. Boekelheide and W. J. Linn. Rearrangements of N-Oxide. Anovel Synthesis of Pridyl Carbinol and Andehydes. J Amer Chem Soc, 76, 1286～1290 (1954)
[31] Thomas L. G, Christl. Heterocyclic Chemistry (third edition) 1997.
[32] R. Bodalski and A. R. Katritzky. New rearrangement types in reactions of acetic anhydride with2-alkylpridine 1-oxides. Tetrahedron letters, 1968,
257(3) 257-260.
[33] Shigeru Oae, Teijiro Kitao and Yoshinori Kitaoka. The Mechanism of the reaction of 2-piconline N-oxide with acetic anhydride. J Amer Chem Soc, 84, 3359(1962)
[34] 刘波，高惠强和周洵钧，合成芳基硫脲的新方法．化学通报 1994，(5)：42-43
[35] R. Jason Herr, J. louise Kuhler, Harold Mecker etc. A Convenient Method for the Preparation of Primary and Symmetrical N, N'-Disubstitued Thioreas. Synthesis 2000. No. 11.1569-1574
[36] C.R. Rasmussen, F. J. Villani, L. E. Weaner. B. Ereynolds etc. Improved Procedures for the Preparation of Cycloalkyl-,Arylalkyl-, and Arythioures. Synthesis, 1988, 6,456-459.
[37] 实用有机化学手册．李述文 范如霖编译 上海科技技术出版社，1986年，168-169
[38] Acros organics Catalogue of Fine Chemicals. International Edition, 2000-2001
[39] 计算机辅助设计---原理、方法及应用．陈凯先，蒋华良，嵇汝运，上海：上海科技技术出版社，2000：165～174