新型肠动力激动剂GK-1806的促肠动力作用及其靶点确认
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摘要
多孔菌D-32 是1980 年一位辽宁省山区果农无意间在一棵桉梨树上发现的一种野生菌,服用该菌对胃肠动力障碍、腹胀、消化不良、慢性便秘等消化道疾病有奇效。国际上对多孔菌的研究结果表明其无毒,可食,性温味甘,能调节机体代谢,增进健康,经常食用对人体机能调节起重要的作用。其子实体中的齿孔酸可用于合成甾体药物,是治疗内分泌疾病,雄性器官衰退及某些妇科疾病的主要药物,也是治疗乳腺癌、前列腺癌等恶性肿瘤的辅助药物。但对于消化道疾病的作用及机制、有效成分、理化性质均未见报道。GK-1806 是一种从多孔菌D-32 中提取出来的肠动力激动剂。其具有很强的水溶性,并且至少可稳定存在于1N 的酸或碱中。本实验以兔离体肠管为靶器官,分别加入不同剂量的GK-1806 和对照药物红霉素来观察其与离体肠管的量效关系,并通过加入神经激肽(NK3)特异性拮抗剂H-2788 来确认GK-1806 的作用靶点,从而确定GK-1806 促肠动力的作用以及其作用靶点,并与红霉素的作用相比较,为肠动力药物筛选奠定了实验基础。
方法:将体外制备的兔离体肠管分别与不同剂量的GK-1806 相互作用,来观察其作用效应,通过同样的方法来观察对照药物红霉素的作用效应。将体外制备的兔离体肠管与NK3受体特异性拮抗剂H-2788 相互作用,来观察其作用效应。NK_3 受体拮抗剂H-2788 与不同
剂量的GK-1806 同时作用于体外制备的兔离体肠管,来观察其作用效应。NK3受体拮抗剂H-2788 与不同剂量的对照药物红霉素同时作用于体外制备的兔离体肠管,来观察其作用效应。同时对体外制备的兔离体肠管的收缩形态及其波形进行观察。
结果:(1)不同剂量的GK-1806 作用于兔离体肠管可出现随剂量增加,效应逐渐增强的典型的S 型曲线,其最大收缩力可达到基础收缩力3.5 倍,并且肠肌不会出现痉挛。(2)不同剂量的对照药物红霉素作用于兔离体肠管亦可出现随剂量增加,效应逐渐增强的典型的S型曲线,其最大收缩力可达到基础收缩力3 倍,当达到最大收缩力时肠肌出现痉挛。(3)NK_3受体拮抗剂H-2788 作用于兔离体肠管,对其收缩效应无明显影响。(4)NK_3受体拮抗剂H-2788 与不同剂量的GK-1806 同时作用于体外制备的兔离体肠管时,GK-1806 对兔离体肠管的收缩效应被强烈的抑制了。(5)NK_3受体拮抗剂H-2788 与不同剂量的对照药物红霉素同时作用于体外制备的兔离体肠管时,其收缩效应与红霉素单独作用于兔离体肠管时一致。
结论:(1)GK-1806 是一种强效的胃肠动力促进剂,其作用机制可能为与内在原发传入性神经元(IPANs)细胞膜表面的NK_3 受体作用,通过肠神经回路引起兔离体肠管的收缩效应。(2)NK_3受体拮抗剂H-2788 对兔离体肠管的收缩效应无明显影响,表明NK_3受体在兔离体肠管的基础收缩状态下不起作用。(3)通过体外制备兔离体肠管的方法,检测GK-1806 的作用,寻找其作用的靶点,从而为筛选胃肠动力药物建立了一种快速、高通量的实验模型。
Polypore D-32, which was growing on the eucalyptus, was discovered by a farmer in Liaoning province. And it has amazing effect on gastrointestinal dyskinesis、abdominal distention、dyspepsia、chronic constipation and etc. The international research on the polypore shows that it is nonpoisonous、edible、sweet and has important function on the regulation of metabolism、improve health、disease resistance、regulation of human’s function. The ehuricoic acid in its fruiting body can be used for synthesis of steroid drug, which is the main drug of endocrine disease、impotence and some diseases of department of obstetrics & gynecology, it could be used for the adjunctive drug of mammary cancer、carcinoma of prostate and other malignant tumor. But its ingredient, which effect on the gastrointestinal tract, has never been reported including action mechanism and physicochemistry character. GK-1806 is a kind of intestinal agonist extracted from the polypore D-32. It is diffluent in water and remain stable in pH 0~14. In this experiment, we utilize the ileal rabbit preparations in vitro as the organ of target, and then add different doses of GK-1806 and its control drug erythrocin to find out their dose-effect relationship. We also add NK3 special antagonist H-2788 to find out its target of motility, then ensure the effect of GK-1806 and its target, compare to erythrocin’s effect. Maybe a new kind of model for intestinal motility drug screening could be
established.
Method: Add different doses of GK-1806 to the ileal rabbit preparations in vitro, and then observe its effect. As the same method observe erythrocin’s effect. Add certain dose of NK3 special antagonist H-2788 to the ileal rabbit preparations in vitro, and then observe its effect. Add the certain dose of NK3 special antagonist H-2788 and different doses of GK-1806 to the ileal rabbit preparations in vitro together, and then observe their collective effect. Add the certain dose of NK3 special antagonist H-2788 and different doses of erythrocin to the ileal rabbit preparations in vitro together, and then observe their collective effect. At the same time, we observe contraction and wave in the computer of the ileal rabbit preparations in vitro going along above steps.
Results: (1) Adding different doses of GK-1806 to the ileal rabbit preparations in vitro, we find out that as the dose increasing the effect increases gradually. Its dose-effect curve is the typical S shape. And its maximal effect value is 3.5 times of basical effect value. The ileal rabbit preparations in vitro never have spasm. (2) As to erythrocin, it also has an increasing effect following the dose increasing. Its dose-effect curve is also typical S shape. And its maximal effect value is 3 times of basical effect value. But when the contraction achieves the maximal effect, the ileal rabbit preparations in vitro have spasm. (3) The NK_3 special antagonist H-2788 has little effect on the ileal rabbit preparations in vitro. (4) Adding different doses of GK-1806 to the ileal rabbit preparations in vitro together with the certain dose of the NK_3 special antagonist H-2788, we can find out that the contraction effect was inhibited strongly. (5) Adding different doses of erythrocin to the ileal rabbit preparations in vitro together with the certain dose of the NK3 special antagonist H-2788, we can find out that the contraction effect was the same as using eryhtrocin alone.
Conclusion: (1) The contractile effect of the ileal rabbit preparations can be promoted potently by GK-1806 potently. Its mechanism may be
react with the NK_3 receptor, and then cause the contractile effect of the ileal rabbit preparations through the enteric nervous system. (2) The NK_3 special antagonist H-2788 has little effect on the ileal rabbit preparations in vitro, which shows that the NK_3 receptor has little reaction at the basical contraction. (3) We can build up a new experiment model of new drug screening, which has high efficiency and high dose through this experiment that GK-1806’s function on the ileal rabbit preparat
方法:将体外制备的兔离体肠管分别与不同剂量的GK-1806 相互作用,来观察其作用效应,通过同样的方法来观察对照药物红霉素的作用效应。将体外制备的兔离体肠管与NK3受体特异性拮抗剂H-2788 相互作用,来观察其作用效应。NK_3 受体拮抗剂H-2788 与不同
剂量的GK-1806 同时作用于体外制备的兔离体肠管,来观察其作用效应。NK3受体拮抗剂H-2788 与不同剂量的对照药物红霉素同时作用于体外制备的兔离体肠管,来观察其作用效应。同时对体外制备的兔离体肠管的收缩形态及其波形进行观察。
结果:(1)不同剂量的GK-1806 作用于兔离体肠管可出现随剂量增加,效应逐渐增强的典型的S 型曲线,其最大收缩力可达到基础收缩力3.5 倍,并且肠肌不会出现痉挛。(2)不同剂量的对照药物红霉素作用于兔离体肠管亦可出现随剂量增加,效应逐渐增强的典型的S型曲线,其最大收缩力可达到基础收缩力3 倍,当达到最大收缩力时肠肌出现痉挛。(3)NK_3受体拮抗剂H-2788 作用于兔离体肠管,对其收缩效应无明显影响。(4)NK_3受体拮抗剂H-2788 与不同剂量的GK-1806 同时作用于体外制备的兔离体肠管时,GK-1806 对兔离体肠管的收缩效应被强烈的抑制了。(5)NK_3受体拮抗剂H-2788 与不同剂量的对照药物红霉素同时作用于体外制备的兔离体肠管时,其收缩效应与红霉素单独作用于兔离体肠管时一致。
结论:(1)GK-1806 是一种强效的胃肠动力促进剂,其作用机制可能为与内在原发传入性神经元(IPANs)细胞膜表面的NK_3 受体作用,通过肠神经回路引起兔离体肠管的收缩效应。(2)NK_3受体拮抗剂H-2788 对兔离体肠管的收缩效应无明显影响,表明NK_3受体在兔离体肠管的基础收缩状态下不起作用。(3)通过体外制备兔离体肠管的方法,检测GK-1806 的作用,寻找其作用的靶点,从而为筛选胃肠动力药物建立了一种快速、高通量的实验模型。
Polypore D-32, which was growing on the eucalyptus, was discovered by a farmer in Liaoning province. And it has amazing effect on gastrointestinal dyskinesis、abdominal distention、dyspepsia、chronic constipation and etc. The international research on the polypore shows that it is nonpoisonous、edible、sweet and has important function on the regulation of metabolism、improve health、disease resistance、regulation of human’s function. The ehuricoic acid in its fruiting body can be used for synthesis of steroid drug, which is the main drug of endocrine disease、impotence and some diseases of department of obstetrics & gynecology, it could be used for the adjunctive drug of mammary cancer、carcinoma of prostate and other malignant tumor. But its ingredient, which effect on the gastrointestinal tract, has never been reported including action mechanism and physicochemistry character. GK-1806 is a kind of intestinal agonist extracted from the polypore D-32. It is diffluent in water and remain stable in pH 0~14. In this experiment, we utilize the ileal rabbit preparations in vitro as the organ of target, and then add different doses of GK-1806 and its control drug erythrocin to find out their dose-effect relationship. We also add NK3 special antagonist H-2788 to find out its target of motility, then ensure the effect of GK-1806 and its target, compare to erythrocin’s effect. Maybe a new kind of model for intestinal motility drug screening could be
established.
Method: Add different doses of GK-1806 to the ileal rabbit preparations in vitro, and then observe its effect. As the same method observe erythrocin’s effect. Add certain dose of NK3 special antagonist H-2788 to the ileal rabbit preparations in vitro, and then observe its effect. Add the certain dose of NK3 special antagonist H-2788 and different doses of GK-1806 to the ileal rabbit preparations in vitro together, and then observe their collective effect. Add the certain dose of NK3 special antagonist H-2788 and different doses of erythrocin to the ileal rabbit preparations in vitro together, and then observe their collective effect. At the same time, we observe contraction and wave in the computer of the ileal rabbit preparations in vitro going along above steps.
Results: (1) Adding different doses of GK-1806 to the ileal rabbit preparations in vitro, we find out that as the dose increasing the effect increases gradually. Its dose-effect curve is the typical S shape. And its maximal effect value is 3.5 times of basical effect value. The ileal rabbit preparations in vitro never have spasm. (2) As to erythrocin, it also has an increasing effect following the dose increasing. Its dose-effect curve is also typical S shape. And its maximal effect value is 3 times of basical effect value. But when the contraction achieves the maximal effect, the ileal rabbit preparations in vitro have spasm. (3) The NK_3 special antagonist H-2788 has little effect on the ileal rabbit preparations in vitro. (4) Adding different doses of GK-1806 to the ileal rabbit preparations in vitro together with the certain dose of the NK_3 special antagonist H-2788, we can find out that the contraction effect was inhibited strongly. (5) Adding different doses of erythrocin to the ileal rabbit preparations in vitro together with the certain dose of the NK3 special antagonist H-2788, we can find out that the contraction effect was the same as using eryhtrocin alone.
Conclusion: (1) The contractile effect of the ileal rabbit preparations can be promoted potently by GK-1806 potently. Its mechanism may be
react with the NK_3 receptor, and then cause the contractile effect of the ileal rabbit preparations through the enteric nervous system. (2) The NK_3 special antagonist H-2788 has little effect on the ileal rabbit preparations in vitro, which shows that the NK_3 receptor has little reaction at the basical contraction. (3) We can build up a new experiment model of new drug screening, which has high efficiency and high dose through this experiment that GK-1806’s function on the ileal rabbit preparat
引文
1. 高爱军付燕任军华. 19例腹部术后胃肠动力障碍浅析. 医学理论与实践,2004, 17: 664.
2. 师笃雷,孙晓红. 腹部手术术后胃肠动力障碍13例分析.华北煤炭医学院学报,2003,5:341
3. 王邦茂, 陈鑫. 干细胞与胃肠动力障碍性疾病的关系研究进展. 基础医学与临床,2003,23:4-5
4. 翟宏丽. 功能性消化不良胃肠动力障碍与胃肠激素的关系. 胃肠病学和肝病学杂志,2003,12:318-21
5. 林连捷,郑长青. 常见胃肠动力障碍性疾病的诊断及治疗. 辽宁药物与临床,2003,6:116-8
6. 孔慧君,郭旭丽. 红霉素持续静滴治疗早产儿胃肠动力障碍29例临床观察. 中国社区医师,2003,19:26
7. 孔慧君,郭旭丽. 红霉素持续静滴治疗早产儿胃肠动力障碍29例临床观察. 中国临床医生,2004,32:35
8. 高军林. 胆道手术后胃肠动力障碍12例诊治体会. 青海医药杂志, 2001,31:28-9
9. 许春娣. 加强对小儿胃肠动力障碍性疾病的临床研究与认识. 临床儿科杂志,2003,21:755-6
10. 宋于刚,孙风蓬. 老年人胃肠动力障碍性疾病研究现状. 第一军医大学报,1999,19:89-91
11. 刘建湘,刘新光. 外科手术后的胃肠动力障碍. 中国实用内科杂志,2001,21:591-3
12. 陆明. 小儿胃肠动力障碍. 临床儿科杂志. 1994,12(3):205-6
13. Furness JB,et al. Intrinsic primary afferent neurons and nerve circuits within the intestine. Progress Neurobio 2004,72: 143–164
14. Riccardo Patacchini, Carlo Alberto Maggi. Peripheral tachykinin receptors as targets for new drugs. E J Pharm. 2001,429:13–21
15. Jocelyn N. Pennefathera,et al. Tachykinins and tachykinin receptors: a growing family Life. Sciences. 2004,74 :1445–63
16. Anthes JC,Chapman RW. SCH 206272: a potent, orally active tachykinin NK1, NK2, and NK3 receptor antagonist. E J Pharm. 2002,450 :191–202
17. Holzer P, Holzer-Petsche U. Tachykinins in the gut: Part II. Roles in neural excitation, secretion and inflammation. Pharmacol. Ther. 1997,73:219–63.
18. Patacchini R, et al. Tachykinin autoreceptors in the gut. Trends Pharmacol. Sci. 2000,21:166.
19. 郭建强,柯美云. 胃肠动力药进展及评价. 中国新药杂志,2001,10: 411-5.
20. 刘思纯. 促胃肠动力药研究现状. 新医学,2003,34:759-60.
21. Furness JB, Sanger GJ. Intrinsic nerve circuits of the gastrointestinal tract:identification of drug targets. Curr Opin Pharm. 2002,2:612–622.
22. Carlo Alberto Maggi. Principles of tachykininergic co-transmission in the peripheral and enteric nervous system. Regu Pep. 2000,93:53–64.
23. AKIHIRO ASAKAWA,et al. Motilin Increases Food Intake in Mice. Peptides. 1998,19:987–990.
24. Fumihiko Sato et al. EM574, an erythromycin derivative, is a motilin receptor agonist in the rabbit. E J Pharm. 1997,322:63–71.
25. P. POITRAS, et al. Heterogeneity of Motilin Receptors in the Gastrointestinal Tract of the Rabbit. Peptides. 1996,17(4):701-707.
2. 师笃雷,孙晓红. 腹部手术术后胃肠动力障碍13例分析.华北煤炭医学院学报,2003,5:341
3. 王邦茂, 陈鑫. 干细胞与胃肠动力障碍性疾病的关系研究进展. 基础医学与临床,2003,23:4-5
4. 翟宏丽. 功能性消化不良胃肠动力障碍与胃肠激素的关系. 胃肠病学和肝病学杂志,2003,12:318-21
5. 林连捷,郑长青. 常见胃肠动力障碍性疾病的诊断及治疗. 辽宁药物与临床,2003,6:116-8
6. 孔慧君,郭旭丽. 红霉素持续静滴治疗早产儿胃肠动力障碍29例临床观察. 中国社区医师,2003,19:26
7. 孔慧君,郭旭丽. 红霉素持续静滴治疗早产儿胃肠动力障碍29例临床观察. 中国临床医生,2004,32:35
8. 高军林. 胆道手术后胃肠动力障碍12例诊治体会. 青海医药杂志, 2001,31:28-9
9. 许春娣. 加强对小儿胃肠动力障碍性疾病的临床研究与认识. 临床儿科杂志,2003,21:755-6
10. 宋于刚,孙风蓬. 老年人胃肠动力障碍性疾病研究现状. 第一军医大学报,1999,19:89-91
11. 刘建湘,刘新光. 外科手术后的胃肠动力障碍. 中国实用内科杂志,2001,21:591-3
12. 陆明. 小儿胃肠动力障碍. 临床儿科杂志. 1994,12(3):205-6
13. Furness JB,et al. Intrinsic primary afferent neurons and nerve circuits within the intestine. Progress Neurobio 2004,72: 143–164
14. Riccardo Patacchini, Carlo Alberto Maggi. Peripheral tachykinin receptors as targets for new drugs. E J Pharm. 2001,429:13–21
15. Jocelyn N. Pennefathera,et al. Tachykinins and tachykinin receptors: a growing family Life. Sciences. 2004,74 :1445–63
16. Anthes JC,Chapman RW. SCH 206272: a potent, orally active tachykinin NK1, NK2, and NK3 receptor antagonist. E J Pharm. 2002,450 :191–202
17. Holzer P, Holzer-Petsche U. Tachykinins in the gut: Part II. Roles in neural excitation, secretion and inflammation. Pharmacol. Ther. 1997,73:219–63.
18. Patacchini R, et al. Tachykinin autoreceptors in the gut. Trends Pharmacol. Sci. 2000,21:166.
19. 郭建强,柯美云. 胃肠动力药进展及评价. 中国新药杂志,2001,10: 411-5.
20. 刘思纯. 促胃肠动力药研究现状. 新医学,2003,34:759-60.
21. Furness JB, Sanger GJ. Intrinsic nerve circuits of the gastrointestinal tract:identification of drug targets. Curr Opin Pharm. 2002,2:612–622.
22. Carlo Alberto Maggi. Principles of tachykininergic co-transmission in the peripheral and enteric nervous system. Regu Pep. 2000,93:53–64.
23. AKIHIRO ASAKAWA,et al. Motilin Increases Food Intake in Mice. Peptides. 1998,19:987–990.
24. Fumihiko Sato et al. EM574, an erythromycin derivative, is a motilin receptor agonist in the rabbit. E J Pharm. 1997,322:63–71.
25. P. POITRAS, et al. Heterogeneity of Motilin Receptors in the Gastrointestinal Tract of the Rabbit. Peptides. 1996,17(4):701-707.
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