手性分子结晶拆分-晶云药物.pdf

1课程八课程八课程八课程八：手性药物分子的结晶提纯手性药物分子的结晶提纯手性药物分子的结晶提纯手性药物分子的结晶提纯晶云药物第一届晶型专题技术培训主讲人主讲人主讲人主讲人：陈敏华博士，首席执行官Crystal Pharmatech苏州晶云药物科技有限公司Email：电话：[phone]Crystal PharmatechCrystal Pharmatech2提纲 手性药物结晶拆分在原料药生产中的重要性 手性药物结晶拆分的原理及工艺研发的流程和策略：三元相图的应用 实例分析:对于不同种类的对映异构体和非对映异构体进行手性拆分不同策略的成功应用Crystal PharmatechCrystal Pharmatech3Enantiomers:stereoisomers which are mirror images of each other(R vs.S)Enantiomeric excess(ee)=(R-S)/(R+S)(0.92 or 92%)Racemate:an equimolar mixture of a pair of enantiomersDiastereomers or diastereoisomers:stereoisomers which are not mirror images of each other(R,S)vs.(R,R)Diastereomeric excess(de)Stereoisomers:mirror image enantiomers,geometric(cis/trans)isomers,and diastereoisomers手性化合物常用术语Crystal PharmatechCrystal Pharmatech4R-SR-SR-SR-SR-SR-SR R R RR R R RR R R R+S S S SS S S SS S S SR S S RS R R SR S R SRacematePhysical mixture of pure enantiomeric crystalsOne phase crystalline additional compoundSolid solution对映异构体分类ConglomerateRacemic CompoundPseudoracemateCrystal PharmatechCrystal Pharmatech5 市场上50%的药物是手性分子 70%的正在研发的药物是手性分子 全球销量最好的10个药物中，9个是手性分子 对于手性药物来说，生产和制备光学纯的对映异构体非常重要，因为不同的对映异构体可能有非常不同的生物活性手性药物的一些事实Crystal PharmatechCrystal Pharmatech6美国药监局对于新的手性药物开发的指南http:/www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm122883.htmCrystal PharmatechCrystal Pharmatech7不同对映异构体具有相同活性的一些手性药物 Both enantiomers of dobutamine are positive inotropes;Both ibuprofen enantiomers are anti-inflammatory agents;Both enantiomers of warfarin and phenprocoumon are anticoagulants;The enantiomers of bupivicaine both produce local anesthesia;http:/www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm122883.htmCrystal PharmatechCrystal Pharmatech8 Granulocytopenia is related to the d-isomer of levodopa;vomiting is caused by the d-isomer of levamisole;and myasthenia gravis symptoms were no longer observed when the d-isomer was removed from d,lcarnitine.不同对映异构体具有不同生物活性的手性药物Crystal PharmatechCrystal Pharmatech9Hypothetical Interaction between the 2 Enantiomers of A Chiral Drug and Its Binding Sitehttp:/ PharmatechCrystal Pharmatech10 Although it is now technologically feasible to prepare purified enantiomers,development of racemates may continue to be appropriate.However,currently available information suggests that the following should be considered in product development:Appropriate manufacturing and control procedures should be used to assure stereoisomeric composition of a product,with respect to identity,strength,quality and purity.Manufacturers should notify compendia of these specifications and tests.Pharmacokinetic evaluations that do not use a chiral assay will be misleading if the disposition of the enantiomers is different.Therefore,techniques to quantify individual stereoisomers in pharmacokinetic samples should be available early.If the pharmacokinetics of the enantiomers are demonstrated to be the some or to exist as a fixed-ratio in the target population,an achiral assay or an assay that monitors one of the enantiomers may be used,subsequently.http:/www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm122883.htm美国药监局对于新的手性药物开发的指南Crystal PharmatechCrystal Pharmatech11Some Examples of Chiral Psychiatric DrugsCrystal PharmatechCrystal Pharmatech12得到纯手性药物的主要途径+chiral resolving agentsDiastereomeric saltsCrystallization/dissolutionSalts with desirable eePure compound in desirable formCompounds(0%ee)Asymmetric synthesisEnantiomeric mixtures(80-96%ee)Crystallization/dissolutionCompounds with desirable eeCrystal PharmatechCrystal Pharmatech13如何进行手性药物结晶提纯工艺的开发？现状现状现状现状 开发工艺耗时太长，很多随机的实验 缺少对手性分子晶型和所属体系的系统了解 对手性结晶提纯的工艺的可重复性和可放大性没有把握，缺少对工艺风险的预测（不知道结晶提纯根据的是动力学还是热力学原理）利用三元相图指导结晶工艺开发利用三元相图指导结晶工艺开发利用三元相图指导结晶工艺开发利用三元相图指导结晶工艺开发 可以全面理解结晶提纯的原理和过程 了解通过结晶工艺提高手性纯度的可能性 确定最佳的结晶工艺条件，预测能够提纯到的最高手性纯度以及对应的产率Crystal PharmatechCrystal Pharmatech14两篇利用三元相图指导手性药物结晶提纯工艺开发的文献Crystal PharmatechCrystal Pharmatech15S0102030405060708090100Solvent0102030405060708090100R0102030405060708090100三元相图的基础知识 Q represents 50%S,20%solvent and 30%R.T represents 80%S,0%solvent and 20%R.Any point on the dashed line will represent a system with same ratio of S to R.QTCrystal PharmatechCrystal Pharmatech16S0102030405060708090100Solvent0102030405060708090100R0102030405060708090100ABCsolid Ssolid RS,R,SolventAdd solventsolid Ssolid RsolutionSeparate the solution and solid phaseQPST用三元相图示意手性分子结晶/溶解提纯的工艺流程Crystal PharmatechCrystal Pharmatech17DSolventUAASE(a)SDUAAE(b)rSolventEAASrAAAASolvent123D(c)UAASrAAAASolvent123D(c)U对映异构体的三元相图ConglomerateRacemic compoundPseudoracemateCrystal PharmatechCrystal Pharmatech18SSolventREConglomerate系统的三元相图2var+=CianceGibbs Phase Rule:C=3Fix Pressure1var+=CianceVariance in GreyFG212Blue solid line represents the saturated solution curveRegion ERS:Both R and S are saturated.Region EFS:R is unsaturated and S is saturated.Region AFEEF:Both R and S are unsaturated.AFCrystal PharmatechCrystal Pharmatech19SSolventRR-SERegion EDS:Both R-S and S are saturated.Region EFS:R-S completely dissolves in solution and S is saturated.Region EDE:R-S is saturated and either R or S is unsaturated.Region AFEEF:Both R-S and S are unsaturated.FDF2var+=CianceGibbs Phase Rule:C=3Fix Pressure1var+=Ciance21122Blue solid line represents the saturated solution curveVariance in GreyAERacemic Compound系统的三元相图Crystal PharmatechCrystal Pharmatech20Develop TPDConglomerateRacemic compoundee of starting solids ee of eutectic pointee of eutectic point minimum ee required for product ee of starting solidsMinimum ee required for product ee of eutectic point ee of starting solids(Enriched in solid)(Enriched in solid)(Enriched in supernatant)XRPD,DSC,TGDetermine the thermodynamically most stable form of racemate?Determine Type of racemate?Solid SolutionMake racemate12364578(a)(b)(c)对映异构体的结晶提纯工艺开发流程Crystal PharmatechCrystal Pharmatech21Conglomerate SystemVmin=)1(210eeUeuDSolventUAASE()11(12100eeeeUDDDVeupureeupure+Yield(at VVmin)=Yield max=0012eeee+(1)(2)(3)M三元相图的深入理解：ConglomerateCrystal PharmatechCrystal Pharmatech22Racemic compound system(when the starting solid has ee higher than that of eutectic)Vmin=)1(210eeUeu)1)(1()1)(1(100maxeueueeeeeeeeYield+=SDSolventUAAEErp2Yield(at VVmin)=(4)(5)(6)Vminrepresents the volume of solvent(expressed in ml of solvent)to be added to one milligram of solid to reach point M.Dpurerepresents the solubility of desired enantiomer in mg of solid per ml of solvent.M()11(12100eeeeUDDDVeupureeupure+Crystal PharmatechCrystal Pharmatech23Racemic compound system(when the starting solid has ee lower than that of eutectic)eueuUDee0Vmax=VDeu+00.5ee0.5Yield(at VVmax)=Yield(at V=Vmax)=)1()(1ee00eeeeeeeueu+(7)(8)(9)Vmaxrepresents the volume of solvent(expressed in ml of solvent)to be added to 1 mg of solid to reach point M.ee0represents the ee of starting solids whose ee is to be upgraded.eeeurepresents the ee of eutectic point.Deuand Ueurepresent the concentration of desired enantiomer and undesired enantiomer(mg of solids per ml of solvent)at the eutectic point respectively.V represents the volume of solvent(in ml)added to one mg of solid.SDSolventUAAEEp1rMCrystal PharmatechCrystal Pharmatech24案例分析Crystal PharmatechCrystal Pharmatech25Desired enantiomer:(S,S)Undesired enantiomer:(R,R)By asymmetric synthesis,API solids with 92-96%ee can be produced,how to upgrade the ee to above 98%?Compound I案例分析1:A racemiccompound system (with high ee for eutectic point)NHCH3OCH3CH3ONFFFClNCrystal PharmatechCrystal Pharmatech26纯对映异构体 vs.消旋体的物理性质Different DSCDifferent XRPDMost likely,the racemate is racemic compound.To be confirmed by solubility ternary phase diagram.Crystal PharmatechCrystal Pharmatech27S/mgR/mg2:3 IPAC:Heptane(v:v)/0.1 ml7.2 mg/ml S7.2 mg/ml REutectic point:98.80%ee297.6 mg/ml S1.79 mg/ml REracemateAee upgrade of Compound I Form BCrystal PharmatechCrystal Pharmatech28Ternary phase diagram of Form B in 2:3 IPAC:Heptane(v:v)at 25 C98.80%ee94%eeP1P298.0%eeCrystal PharmatechCrystal Pharmatech29Based on the above equations,given a starting mixture with ee of 94%,Vmaxis 0.00318 and yield is 97.6%at V=Vmax.This means by adding 0.00318 ml of 2:3(v:v)IPAC:Heptane solvent to 1 mg of starting enantiomeric mixture with 94%ee at 25.0 C,at equilibrium,the ee will be upgraded to 98.80%with a yield 97.6%.eeUDee)()(0=0.00338 ee0Vmax=V+00.5ee0.56.297Yield VVmax=Optimal process to upgrade ee in 2:3 IPAC:Heptane(v:v)at 25 CFeasibility and strategy:The ee of eutectic point(98.80%)is higher than that of starting solids(94%-96%).Therefore,the ee can be upgraded to a maximum value of 98.80%in the supernatant by dissolution.Determine the optimal solvent/solid ratio and the corresponding yield:Crystal PharmatechCrystal Pharmatech30Chiral amide(penultimate)API92-96%ee92-96%eeDissolutionFilterrecrystallizationAPI 98%eeReactioncrystallizationReactioncrystallization(ee upgrade by dissolution of API)Chiral amide(penultimate)API99%ee99%eeReactioncrystallizationReactioncrystallization(ee upgrade by crystallization of penultimate)ee upgrade in chiral amide crystallization step was achieved when a new anhydrous crystalline form of chiral amide enantiomer was discovered.Prior to the discovery of the anhydrous crystalline form,chiral amide crystallized as a mixture of MTBE solvate and amorphous,with no ee upgrade.New ee upgrade process was implemented in the manufacturing process.中间体新晶型的发现改变了手性分子的结晶提纯工艺Crystal PharmatechCrystal Pharmatech31 D(mg)0 10 20 30 40 50 60 70 80 90100Ethanol(0.1 ml)0102030405060708090100U(mg)0102030405060708090100Composition of total systemComposition of SupernatantED(mg)0 10 20 30 40 50 60 70 80 90100Ethanol(0.1 ml)0102030405060708090100U(mg)0102030405060708090100Composition of total systemComposition of SupernatantEee upgrade of Compound II DIPA Salt案例分析2：A racemic compound system(with low ee for eutectic point)Eutectic point:ee:30.3%C(R):5.36 mg/ml RC(S):2.87 mg/ml SC(R):4.76 mg/mlTemp:25 CNSOOOFClCH3O-Crystal PharmatechCrystal Pharmatech32)1(210eeUeuFeasibility and strategy:ee of eutectic point is 30.3%,lower than that of starting solids(92%to 96%ee).Therefore,ee can be upgraded to 100%in the solid phase by adding Vminor more amount of solvents to the solids.=0.174(1-ee0)Based on above,given a starting enantiomeric mixture with 94%ee,Vmincan be determined to be 0.010 and the yield is 94.4%at V=Vmin.This means that by adding 0.010 ml of EtOH to 1 mg of starting enantiomeric mixture with 94%ee at 25.0 C,at equilibrium,the ee of D will be upgraded to 100%in the solid phase with a yield of 94.4%.Vmin=Optimal process to upgrade ee of DIPA Salt in ethanol at 25 C)1)(1()1)(1(100maxeueueeeeeeeeYield+=Determine the optimal solvent/solid ratio and the corresponding yield:Crystal PharmatechCrystal Pharmatech33案例分析三:A conglomerate Systemee upgrade of compound III freebase D(mg)010 20 30 40 50 60 70 80 90 1001:3(v:v)IPA:Heptane(0.1 ml)0102030405060708090100U(mg)0102030405060708090100Composition of supernatantD(mg)010 20 30 40 50 60 70 80 90 1001:3(v:v)IPA:Heptane(0.1 ml)0102030405060708090100U(mg)0102030405060708090100Composition of supernatant25.0 C45.0 CC(R)=5.6 mgat 25 CEutectic point:C(S)=6.0 mg/mlC(R)=6.0 mg/mlC(R)=14.0 mgEutectic point:C(S)=19.1 mg/mlC(R)=19.1mg/mlNH2NONNNFFFFFFCrystal PharmatechCrystal Pharmatech34ee can be upgraded to 100%in the solid phase by adding Vminor more solvents to the starting solid.Based on the TPD at 45 C,the solubility of pure conglomerate is 14.0 mg of R and 14.0 mg of S per ml of solvent and that of pure enantiomer R is 10.4 mg per ml of solvent.Therefore,similar Vminand yield at VVmincan be calculated based on equations(7)and(8).)1(210eeUeuVmin=0.113(1-ee0)Yield max=0012eeee+Optimal process to upgrade ee of Compound III freebase in 1:3(v:v)IPA:Heptane at 25 C or 45 CFeasibility and strategy:Determine the optimal solvent/solid ratio and the corresponding yield:Crystal PharmatechCrystal Pharmatech35DSolventUE212非对映异构体的三元相图2var+=CianceGibbs Phase Rule:C=3Fix Pressure1var+=CianceVariance in BoldThe ternary phase diagram is not symmetric due to different solubility of two diastereomers.Crystal PharmatechCrystal Pharmatech36Vmin=)1(210deUeu()11(12100dedeUDDDVeupureeupure+Yield(at VVmin)=)1)(1()1)(1(100eueudededede+Yield(at V=Vmin)=Yield is defined as the ratio of desired diastereomer in the solid phase to the desired diastereomer in the total system.de represents the diastereomeric excess.Define the optimal ratio and predict the yield based on the ternary phase diagramCrystal PharmatechCrystal Pharmatech37OHNH2OHNH2(1R,2S)-(+)-cis-(1S,2R)-(-)-cis-MW=149.19Compound IV APICAIFour possible isomers of compound IV CAI saltAPI(R,R,R)-CAI(R,S):D-dAPI(S,S,S)-CAI(R,S):U-dAPI(R,R,R)-CAI(S,R):D-uAPI(S,S,S)-CAI(S,R):U-uAssuming the crude free acid has ee 92%and CAI salt has ee 99%,then upon salt formation,there will be:D-d:95.52%U-d:3.98%D-u:0.48%U-u:0.02%API has three chiral centers.案例分析四:非对映异构体的手性提纯Crystal PharmatechCrystal Pharmatech38Ternary PlotD-d/mg0102030405060708090100Solvent(12.6:87.4(v:v)toluene:heptane plus various impurities)/ml0102030405060708090100U-d/mg0102030405060708090100Total compositionSupernatantMother liquor from processEutecticC(D-d):1.0 mg/mlC(U-d):2.07 mg/mlde:-34.9%Pure D-d:C(D-d)0.92 mg/mlTemperature:23.0 CStarting Solvent:Washed mother liquor(22.8%de,2.02 g/l)Crystal PharmatechCrystal Pharmatech39Ternary PlotD-d(mg)0102030405060708090100Solvent from ML(237573-251)(0.1 ml)0102030405060708090100U-d(mg)0102030405060708090100Supernatant compositionTotal systemEutectic:D-d:19.0 mg/mlU-d:37.7 mg/mlde:-33.04%Starting solvent:Mother liquor(61.5%de,17.7 g/l)Temperature:23.0 CCrystal PharmatechCrystal Pharmatech40 Eutectic de of the diastereomeric system is relatively independent of the solvent composition/impurity concentration and it is very low -34%.Low eutectic de suggests thermodynamically the undesired diastereomer can be rejected in the supernatant and pure desired diastereomer can be obtained in the solids upon crystallization.The optimal solvent/solid ratio and the yield can be pre-defined and calculated.通过结晶提纯的可行性和策略Crystal PharmatechCrystal Pharmatech41总结三元相图是理解手性药物结晶提纯工艺的强有力的工具。根据对映异构体或者非对映异构体系统的三元相图，一个手性药物分子通过结晶或者溶解的方法得到提纯的可行性可以立刻确定，同时三元相图可以协助我们找到最优化的结晶工艺条件。手性药物的手性提纯可以在原料药最后一步的结晶工艺中实现，也可以通过中间体的结晶工艺实现。有时通过对手性药物中间体的结晶，可能可以将一个很难提纯的手性药物通过很简单的方法得以提纯。手性药物的结晶提纯有时会受到结晶动力学的很大影响，不要忽略。Crystal PharmatechCrystal Pharmatech42晶云药物晶云药物晶云药物晶云药物可以帮助您开发出手性药物结晶提纯手性药物结晶提纯手性药物结晶提纯手性药物结晶提纯的工艺，如果您有这方面的任何问题，请随时和我们联系咨询！苏州晶云药物科技有限公司Email：电话：[phone]Crystal PharmatechCrystal Pharmatech43药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息药物晶型研究和药物固态信息研发领域专业研发领域专业研发领域专业研发领域专业研发领域专业研发领域专业研发领域专业研发领域专业CROCROCROCROCROCROCROCRO晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司晶云药物科技有限公司Crystal Pharmatech IncCrystal Pharmatech IncCrystal PharmatechCrystal Pharmatech44公司使命公司使命公司使命公司使命公司使命公司使命公司使命公司使命?为全球各制药公司提供药物晶型研药物晶型研药物晶型研药物晶型研究究究究和药物固态研发药物固态研发药物固态研发药物固态研发领域的最佳技术方案，并成为该领域的领导者。Crystal PharmatechCrystal Pharmatech45公司简介公司简介公司简介公司简介公司简介公司简介公司简