手性固定相分離技術(shù)在藥物研發(fā)中的應(yīng)用PPT

1、目 錄錄 手性藥物分離的重要性手性藥物分離的重要性 測定與獲得單一對映體的方法測定與獲得單一對映體的方法 手性分離原理手性分離原理 在藥物研發(fā)中解決的問題在藥物研發(fā)中解決的問題鏡平面鏡平面對映體對映體 A A對映體對映體 B B手性分子 手性藥物分離的重要性手性藥物分離的重要性Chirality & Its ConsequenceBuilding blocks of life -amino acids, sugars, enzyme.(origin of life)Mirror ImageMirror ImageDrugs, agrochemicals, food additives, fra

2、grances, archeology, ecophysiology, etc.Consequence:Chiral molecules in biological materials Most enzymes are chiral reagents because their binding site is chiral Many drug molecules are synthesized as racemates Either both molecules have comparable activity or one is inactive.生命的手性之分生命的手性之分 作為生命的基本

3、結(jié)構(gòu)單元，氨基酸也有手性作為生命的基本結(jié)構(gòu)單元，氨基酸也有手性之分。驚人的發(fā)現(xiàn)之分。驚人的發(fā)現(xiàn)-組成地球生命體的幾乎組成地球生命體的幾乎都是左旋氨基酸，而沒有右旋氨基酸都是左旋氨基酸，而沒有右旋氨基酸某些右旋分子是人體生命的克星某些右旋分子是人體生命的克星!人是由左旋氨基酸組成的生命體，它不能很人是由左旋氨基酸組成的生命體，它不能很好地代謝右旋分子，在一些含有右旋分子的藥好地代謝右旋分子，在一些含有右旋分子的藥物就會成為負擔，甚至造成對生命體的損害。物就會成為負擔，甚至造成對生命體的損害。表 一些具有不同手性的藥物的藥效差別測定與獲得單一對映體的方法測定與獲得單一對映體的方法不對稱合成法不對稱

4、合成法外消旋體拆分法外消旋體拆分法 手性源合成法手性源合成法色譜拆分法色譜拆分法 化學(xué)拆分法化學(xué)拆分法酶或生物法酶或生物法 高效毛細管電泳高效毛細管電泳氣相色譜氣相色譜(GC)薄層色譜法薄層色譜法超臨界流體色譜超臨界流體色譜(SFC)高效液相色譜法高效液相色譜法 測定與獲得單一對映體的方法測定與獲得單一對映體的方法高效液相色譜高效液相色譜Introduction of chromatographyThe most widely used analytical separations techniqueUtilizes a liquid mobile phase to separate comp

5、onents of mixtureUses high pressure to push solvent through the columnThe developer of chromatographyHigh performance liquid chromatographyHigher resolution and speed of analysisHPLC columns can be reused without repacking or regenerationGreater reproducibility due to close control of the parameters

6、 affecting the efficiency of separationEasy automation of instrument operation and data analysisAdaptability to large-scale, preparative proceduresHPLC的儀器配置及流程的儀器配置及流程 色譜泵自動進樣器 檢測器廢液數(shù)據(jù)處理系統(tǒng)色譜圖：色譜圖：HPLC圖形圖形結(jié)果（結(jié)果（Chromatogram）色譜圖即色譜柱流出物通過檢測器時所產(chǎn)生的響應(yīng)信號對時間的曲線圖，其縱坐標為信號強度，橫坐標為保留時間。 色譜峰保留時間（分）基線峰高峰寬響應(yīng)值色譜分析法分

7、類色譜分析法分類通用通用LC固定相固定相1.通用HPLC固定相填料通用通用LC固定相固定相Packing column and monolithic columnsPacked column A porous particle PolymermonolithSilica monolith液相色譜原理液相色譜原理 基本概念及方法開發(fā)基本概念及方法開發(fā)環(huán)糊精及其衍生物環(huán)糊精及其衍生物3m（Kromasil）掃描電鏡圖 5m（Kromasil）掃描電鏡圖 硅硅膠膠外外消消旋旋體體8000 psi 8000 psi 高壓填裝高壓填裝RuPPh3+PPh3+ClNNH2NSRuPPh3+PPh3+ClN

8、SNH2NOOCRuPh3P+Ph3P+ClNH2NNSCOOOOCRuPh3P+Ph3P+ClNSNNH2COON-N =NN*RuPPh3+PPh3+ClNNH2NSRuPPh3+PPh3+ClNSNH2NOOCRuPh3P+Ph3P+ClNH2NNSCOOOOCRuPh3P+Ph3P+ClNSNNH2COO*RSRRRSSS(a) (b)研創(chuàng)SCDP102520型手性制備柱HPLC色譜圖 分子印跡聚合物（MIP）概概述分子識別設(shè) 想首次合成發(fā) 展酶/底物 抗原/抗體 激素/受體Pauling 抗原為模板合成抗體，1940Wulff, MIP, 1972Mosbach, Nature ,1

9、993What are MIPs?Synthetic polymers with well-defined surface cavities created by templateMIP can recognise the template in presence of similar moleculesu Shape selectivityu Binding groups on cavity surfaceApplications in separation science, catalysis, organic chemistry, immunoassays, sensors,分子印跡技術(shù)

10、術(shù)的基本原理 PrearrangementPolymerizationExtractionMolecular imprinting technologyMosbach(The first user of non-covalent methodology) Pauling(The first developer of MIP) Characteristics of MIP:PredeterminationSpecific recognitionPracticability MIP的制備的制備模板分子、功能單體、交聯(lián)劑、聚合反應(yīng)Nature Biotechnology,2002,20,884Com

11、mon monomers and crosslinkers in MIPCH2OHOacrylic acidCH2OHOmethacrylic acidH3CCH2OCH3Omethyl methacrylic acidH3COHOH2Cp-vinylbenzoic acidOH2COOHOHitaconic acidCH3H2C4-ethystyreneH2C4-ethystyreneNH2CNH2C4-vinylpyridineNNH2C2-vinylpyridine1-vinylimidazoleCH2SO3HCH3CH3CH2Oacrylamido-2-methyl-1-propane

12、- sulphonic acidNH2OH2CNH2OH2CH3CacrylamidemethacrylamideH2CNHOOstyrenetrans-3-(3-pyridyl)-acrylic acidCH2HNHNCH2OON,N-1,4-phenylenediacrylamineCH2HNHNCH2OON,N-methylenediacrylamideHNCH2OCH2O3,5-bis(acryloylamido)benzoic acidHNCO2HCH2H3COOCH3OOOethylene glycol dimethacrylateCH2H2CdivinylbenzeneCH2OH

13、NOOCH2N,O-bisacryloyl-phenylalaninolCH3CH2H3CH2C1,3-diisopropenyl benzeneCH3H2COOCH2OOCH2tetramethylene dimethacrylateNHNCH2HNH2COO2,6-bisacryloylamidopyridineNNOCH2OH2C1,4-diacryloyl piperazine聚合方式本體聚合懸浮聚合乳液聚合溶漲聚合原位聚合表面聚合最簡單的制備方法制備球形MIP產(chǎn)率最高適合作為色譜介質(zhì)制備成本高球形MIP均勻性好在硅膠基質(zhì)上Preparation of imprinted polyme

14、r Preparation of imprinted polymer MIP - for what purpose? MIPapplicationsensor Membrane separationSPEChiral separationPharmic analysis Chromatographic separation artificial antibodies Enzyme catalysisRuPPh3+PPh3+ClNNH2NSRuPPh3+PPh3+ClNSNH2NOOCRuPh3P+Ph3P+ClNH2NNSCOOOOCRuPh3P+Ph3P+ClNSNNH2COON-N =NN*RuPPh3+PPh3+ClNNH2NSRuPPh3+PPh3+ClNSNH2NOOCRuPh3P+Ph3P+ClNH