盐酸吡格列酮与胃蛋白酶之间的相互作用及对药效、胃部消化能力的影响
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摘要
为了研究药物与胃蛋白酶(PEP)相互作用后对药效及胃部消化能力的影响,在模拟生理条件下,建立了光谱测定。本文通过荧光法、同步荧光法和分子对接模拟技术研究了在298,310,318 K时盐酸吡格列酮(PGH)和PEP之间的相互作用机理。结果表明,PGH与PEP之间通过静电引力及氢键作用以静态猝灭的方式形成了1∶1的稳定复合物;PEP的酪氨酸残基(Tyr)和色氨酸残基(Trp)均参与了反应;PGH-PEP的结合对后继配体存在正协同作用。通过估算得知:当患者服用PGH 15～45 mg时,胃液中的药物结合率为0.0013%～0.0032%,数值很小,即PGH与PEP的结合对PGH药效几乎没有影响;PEP的结合率为69.76%～87.37%,即服用PGH使得游离的PEP减少69.76%～87.37%,表明服用PGH将使患者的消化功能受到影响。分子对接技术表明PGH与PEP的最佳结合位点位于PEP的催化活性中心处,且两者的结合作用改变了PEP催化活性中心处氨基酸残基的微环境。
In order to study the effect of the interaction between drugs and pepsin(PEP) on the pharmacodynamics and digestive ability of stomach, the spectral determination was established under simulated physiological conditions. At 298, 310 and 318 K, the interaction mechanism between pioglitazone hydrochloride(PGH) and PEP was studied by fluorescence, synchronous fluorescence and molecular docking simulation. The results showed that a stable 1∶1 complex was formed between PGH and PEP by electrostatic force and hydrogen bonding in a static quenching manner. Tyrosine residues(Tyr) and tryptophan residues(Trp) in PEP were involved in the reaction. The binding of PGH-PEP had positive cooperativity effect on the subsequent ligands. The results showed that when patients took PGH 15-45 mg, the drug binding rate in gastric juice was 0.0013%-0.0032%, which was very small, that is, the combination of PGH and PEP had little effect on PGH. The binding rate of PEP was 69.76%-87.37%, that is, PGH reduced the free PEP by 69.76%-87.37%, indicating that PGH would affect the digestive function of the patients. Molecular docking technique showed that the best binding site between PGH and PEP was located at the catalytic active site of PEP, and the interaction between them changed the microenvironment of amino acid residues in PEP catalytic active center.
In order to study the effect of the interaction between drugs and pepsin(PEP) on the pharmacodynamics and digestive ability of stomach, the spectral determination was established under simulated physiological conditions. At 298, 310 and 318 K, the interaction mechanism between pioglitazone hydrochloride(PGH) and PEP was studied by fluorescence, synchronous fluorescence and molecular docking simulation. The results showed that a stable 1∶1 complex was formed between PGH and PEP by electrostatic force and hydrogen bonding in a static quenching manner. Tyrosine residues(Tyr) and tryptophan residues(Trp) in PEP were involved in the reaction. The binding of PGH-PEP had positive cooperativity effect on the subsequent ligands. The results showed that when patients took PGH 15-45 mg, the drug binding rate in gastric juice was 0.0013%-0.0032%, which was very small, that is, the combination of PGH and PEP had little effect on PGH. The binding rate of PEP was 69.76%-87.37%, that is, PGH reduced the free PEP by 69.76%-87.37%, indicating that PGH would affect the digestive function of the patients. Molecular docking technique showed that the best binding site between PGH and PEP was located at the catalytic active site of PEP, and the interaction between them changed the microenvironment of amino acid residues in PEP catalytic active center.
引文
[1] ZHANG C C,MATZGER A J.A newly discovered racemic compound of pioglitazone hydrochloride is more stable than the commercial conglomerate [J].Cryst.Growth Des.,2017,17(2):414-417.
[2] 王静文,曹进,王钢力,等.保健食品中非法添加药物检测技术研究进展 [J].药物分析杂志,2014,34(1):1-11.WANG J W,CAO J,WANG G L,et al..Research progress on determination technologies for illegally added drugs in health food [J].Chin.J.Pharm.Anal.,2014,34(1):1-11.(in Chinese)
[3] 李改霞,刘保生,李志云,等.盐酸吡格列酮与BSA反应机理的荧光猝灭法、同步荧光法比较研究 [J].发光学报,2015,36(6):705-710.LI G X,LIU B S,LI Z Y,et al..Comparative studies on the reaction mechanism of pioglitazone hydrochloride with bovine serum albumin by fluorescence spectroscopy and synchronous fluorescence spectroscopy [J].Chin.J.Lumin.,2015,36(6):705-710.(in Chinese)
[4] WANG J,CHAN C,HUANG F W,et al..Interaction mechanism of pepsin with a natural inhibitor gastrodin studied by spectroscopic methods and molecular docking [J].Med.Chem.Res.,2017,26(2):405-413.
[5] YING M,HUANG F W,YE H D,et al..Study on interaction between curcumin and pepsin by spectroscopic and docking methods [J].Int.J.Biol.Macromol.,2015,79:201-208.
[6] MA X L,HE J W,HUANG Y M,et al..Investigation and comparison of the binding between tolvaptan and pepsin and trypsin:multi-spectroscopic approaches and molecular docking [J].J.Mol.Recognit.,2017,30(5):e2598-1-9.
[7] G?KOGLU E,YILMAZ E.Fluorescence interaction and determination of sulfathiazole with trypsin [J].J.Fluoresc.,2014,24(5):1439-1445.
[8] TAYYAB S,IZZUDIN M M,KABIR M Z,et al..Binding of an anticancer drug,axitinib to human serum albumin:fluorescence quenching and molecular docking study [J].J.Photochem.Photobiol.B,2016,162:386-394.
[9] RAZA M,YANG J,YUN W,et al..Insights from spectroscopic and in-silico techniques for the exploitation of biomolecular interactions between human serum albumin and paromomycin [J].Colloids Surf.B Biointerfaces,2017,157:242-253.
[10] MOEINPOUR F,MOHSENI-SHAHRI F S,MALAEKEH-NIKOUEI B,et al..Investigation into the interaction of losartan with human serum albumin and glycated human serum albumin by spectroscopic and molecular dynamics simulation techniques:a comparison study [J].Chem.Biol.Interact.,2016,257:4-13.
[11] FEROZ S R,TEOH Y J,MOHAMAD S B,et al..Interaction of flavokawain B with lysozyme:a photophysical and molecular simulation study [J].J.Lumin.,2015,160:101-109.
[12] AMROABADI M K,TAHERI-KAFRANI A,SAREMI L H,et al..Spectroscopic studies of the interaction between alprazolam and Apo-human serum transferrin as a drug carrier protein [J].Int.J.Biol.Macromol.,2018,108:263-271.
[13] BANI-YASEEN A D.Spectrofluorimetric study on the interaction between antimicrobial drug sulfamethazine and bovine serum albumin [J].J.Lumin.,2011,131(5):1042-1047.
[14] BUDDANAVAR A T,NANDIBEWOOR S T.Multi-spectroscopic characterization of bovine serum albumin upon interaction with atomoxetine [J].J.Pharm.Anal.,2017,7(3):148-155.
[15] EHTESHAMI M,RASOULZADEH F,MAHBOOB S,et al..Characterization of 6-mercaptopurine binding to bovine serum albumin and its displacement from the binding sites by quercetin and rutin [J].J.Lumin.,2013,135:164-169.
[16] XU C B,GU J L,MA X P,et al..Investigation on the interaction of pyrene with bovine serum albumin using spectroscopic methods [J].Spectrochim.Acta A Mol.Biomol.Spectrosc.,2014,125:391-395.
[17] 赵旭,王毅力,郭瑾珑,等.颗粒物微界面吸附模型的分形修正——朗格缪尔(Langmuir)、弗伦德利希(Freundlich)和表面络合模型 [J].环境科学学报,2005,25(1):52-57.ZHAO X,WANG Y L,GUO J L,et al..Modification of the micro-interface adsorption model on particles with fractal theory-Langmuir,Freundlich and surface complexation adsorption model [J].Acta Sci.Circumstant.,2005,25(1):52-57.(in Chinese)
[18] 冯军鹏,陈秀,王瑞玲,等.不同胃液中胃蛋白酶含量的测定 [J].中国医学工程,2012,20(3):3-5.FENG J P,CHEN X,WANG R L,et al..Assay of the pepsin content in the various gastric juice [J].China Med.Eng.,2012,20(3):3-5.(in Chinese)
[19] HUANG S,QIU H N,LIU Y,et al..Systematical investigation of in vitro interaction of InP/ZnS quantum dots with human serum albumin by multispectroscopic approach [J].Colloids Surf.B Biointerfaces,2016,148:165-172.
[20] BOLATTIN M B,NANDIBEWOOR S T,JOSHI S D,et al..Interaction of hydralazine with human serum albumin and effect of β-cyclodextrin on binding:insights from spectroscopic and molecular docking techniques [J].Ind.Eng.Chem.Res.,2016,55(19):5454-5464.
[21] HU Y J,YANG Y O,DAI C M,et al..Site-selective binding of human serum albumin by palmatine:spectroscopic approach [J].Biomacromolecules,2010,11(1):106-112.
[22] MARIAM J,DONGRE P M,KOTHARI D C.Study of interaction of silver nanoparticles with bovine serum albumin using fluorescence spectroscopy [J].J.Fluoresc.,2011,21(6):2193-2199.
[23] TIAN Z Y,ZANG F L,LUO W,et al..Spectroscopic study on the interaction between mononaphthalimide spermidine (MINS) and bovine serum albumin (BSA) [J].J.Photochem.Photobiol.B,2015,142:103-109.
[24] CUI M M,LIU B S,LI T T,et al..Interaction of transferrin with cefuroxime sodium [J].Spectrosc.Lett.,2016,49(9):573-581.
[25] ZHANG H M,CAO J,FEI Z H,et al..Investigation on the interaction behavior between bisphenol a and pepsin by spectral and docking studies [J].J.Mol.Struct.,2012,1021:34-39.
[2] 王静文,曹进,王钢力,等.保健食品中非法添加药物检测技术研究进展 [J].药物分析杂志,2014,34(1):1-11.WANG J W,CAO J,WANG G L,et al..Research progress on determination technologies for illegally added drugs in health food [J].Chin.J.Pharm.Anal.,2014,34(1):1-11.(in Chinese)
[3] 李改霞,刘保生,李志云,等.盐酸吡格列酮与BSA反应机理的荧光猝灭法、同步荧光法比较研究 [J].发光学报,2015,36(6):705-710.LI G X,LIU B S,LI Z Y,et al..Comparative studies on the reaction mechanism of pioglitazone hydrochloride with bovine serum albumin by fluorescence spectroscopy and synchronous fluorescence spectroscopy [J].Chin.J.Lumin.,2015,36(6):705-710.(in Chinese)
[4] WANG J,CHAN C,HUANG F W,et al..Interaction mechanism of pepsin with a natural inhibitor gastrodin studied by spectroscopic methods and molecular docking [J].Med.Chem.Res.,2017,26(2):405-413.
[5] YING M,HUANG F W,YE H D,et al..Study on interaction between curcumin and pepsin by spectroscopic and docking methods [J].Int.J.Biol.Macromol.,2015,79:201-208.
[6] MA X L,HE J W,HUANG Y M,et al..Investigation and comparison of the binding between tolvaptan and pepsin and trypsin:multi-spectroscopic approaches and molecular docking [J].J.Mol.Recognit.,2017,30(5):e2598-1-9.
[7] G?KOGLU E,YILMAZ E.Fluorescence interaction and determination of sulfathiazole with trypsin [J].J.Fluoresc.,2014,24(5):1439-1445.
[8] TAYYAB S,IZZUDIN M M,KABIR M Z,et al..Binding of an anticancer drug,axitinib to human serum albumin:fluorescence quenching and molecular docking study [J].J.Photochem.Photobiol.B,2016,162:386-394.
[9] RAZA M,YANG J,YUN W,et al..Insights from spectroscopic and in-silico techniques for the exploitation of biomolecular interactions between human serum albumin and paromomycin [J].Colloids Surf.B Biointerfaces,2017,157:242-253.
[10] MOEINPOUR F,MOHSENI-SHAHRI F S,MALAEKEH-NIKOUEI B,et al..Investigation into the interaction of losartan with human serum albumin and glycated human serum albumin by spectroscopic and molecular dynamics simulation techniques:a comparison study [J].Chem.Biol.Interact.,2016,257:4-13.
[11] FEROZ S R,TEOH Y J,MOHAMAD S B,et al..Interaction of flavokawain B with lysozyme:a photophysical and molecular simulation study [J].J.Lumin.,2015,160:101-109.
[12] AMROABADI M K,TAHERI-KAFRANI A,SAREMI L H,et al..Spectroscopic studies of the interaction between alprazolam and Apo-human serum transferrin as a drug carrier protein [J].Int.J.Biol.Macromol.,2018,108:263-271.
[13] BANI-YASEEN A D.Spectrofluorimetric study on the interaction between antimicrobial drug sulfamethazine and bovine serum albumin [J].J.Lumin.,2011,131(5):1042-1047.
[14] BUDDANAVAR A T,NANDIBEWOOR S T.Multi-spectroscopic characterization of bovine serum albumin upon interaction with atomoxetine [J].J.Pharm.Anal.,2017,7(3):148-155.
[15] EHTESHAMI M,RASOULZADEH F,MAHBOOB S,et al..Characterization of 6-mercaptopurine binding to bovine serum albumin and its displacement from the binding sites by quercetin and rutin [J].J.Lumin.,2013,135:164-169.
[16] XU C B,GU J L,MA X P,et al..Investigation on the interaction of pyrene with bovine serum albumin using spectroscopic methods [J].Spectrochim.Acta A Mol.Biomol.Spectrosc.,2014,125:391-395.
[17] 赵旭,王毅力,郭瑾珑,等.颗粒物微界面吸附模型的分形修正——朗格缪尔(Langmuir)、弗伦德利希(Freundlich)和表面络合模型 [J].环境科学学报,2005,25(1):52-57.ZHAO X,WANG Y L,GUO J L,et al..Modification of the micro-interface adsorption model on particles with fractal theory-Langmuir,Freundlich and surface complexation adsorption model [J].Acta Sci.Circumstant.,2005,25(1):52-57.(in Chinese)
[18] 冯军鹏,陈秀,王瑞玲,等.不同胃液中胃蛋白酶含量的测定 [J].中国医学工程,2012,20(3):3-5.FENG J P,CHEN X,WANG R L,et al..Assay of the pepsin content in the various gastric juice [J].China Med.Eng.,2012,20(3):3-5.(in Chinese)
[19] HUANG S,QIU H N,LIU Y,et al..Systematical investigation of in vitro interaction of InP/ZnS quantum dots with human serum albumin by multispectroscopic approach [J].Colloids Surf.B Biointerfaces,2016,148:165-172.
[20] BOLATTIN M B,NANDIBEWOOR S T,JOSHI S D,et al..Interaction of hydralazine with human serum albumin and effect of β-cyclodextrin on binding:insights from spectroscopic and molecular docking techniques [J].Ind.Eng.Chem.Res.,2016,55(19):5454-5464.
[21] HU Y J,YANG Y O,DAI C M,et al..Site-selective binding of human serum albumin by palmatine:spectroscopic approach [J].Biomacromolecules,2010,11(1):106-112.
[22] MARIAM J,DONGRE P M,KOTHARI D C.Study of interaction of silver nanoparticles with bovine serum albumin using fluorescence spectroscopy [J].J.Fluoresc.,2011,21(6):2193-2199.
[23] TIAN Z Y,ZANG F L,LUO W,et al..Spectroscopic study on the interaction between mononaphthalimide spermidine (MINS) and bovine serum albumin (BSA) [J].J.Photochem.Photobiol.B,2015,142:103-109.
[24] CUI M M,LIU B S,LI T T,et al..Interaction of transferrin with cefuroxime sodium [J].Spectrosc.Lett.,2016,49(9):573-581.
[25] ZHANG H M,CAO J,FEI Z H,et al..Investigation on the interaction behavior between bisphenol a and pepsin by spectral and docking studies [J].J.Mol.Struct.,2012,1021:34-39.