Design and Synthesis of Handles for Solid-Phase Peptide Synthesis And
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Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2003 Design and synthesis of handles for solid-phase peptide synthesis and convergent peptide synthesis Jose Giraldes Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Chemistry Commons Recommended Citation Giraldes, Jose, "Design and synthesis of handles for solid-phase peptide synthesis and convergent peptide synthesis" (2003). LSU Doctoral Dissertations. 1146. https://digitalcommons.lsu.edu/gradschool_dissertations/1146 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. DESIGN AND SYNTHESIS OF HANDLES FOR SOLID-PHASE PEPTIDE SYNTHESIS AND CONVERGENT PEPTIDE SYNTHESIS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Chemistry by José Giraldés B.S., University of Puerto Rico, 1997 May, 2003 To my family ii ACKNOWLEDGMENTS I would like to thank my major professor Dr. Mark McLaughlin for his invaluable guidance and support during my stay at LSU. I am very grateful for the freedom and encouragement he gave me to develop my own ideas. A major amount of thanks must be given to Dr. Frank Zhou for the magic angle spinning NMR, to Martha Juban for help with peptide synthesis and purification, to Dr. Frank Fronczek for crystal structure determinations and to Dr. Tracy McCarley and Ms. Renee Sims for mass specs. I also greatly appreciate the scientific discussions with many former current students and post-docs in our laboratory work: Dr. Umut Oguz, Mr. Caleb Clark, Dr. Tanaji Talele, Dr. Mohanraj Kumar, Dr. Ted Gauthier and Dr. Lars Hammarström.. I would also like to thank my friends for their kindness and support: Mr. Leonardo Baez, Dr. Alfonso Davila, Dr. Rachel Bolzan and Dr. Robert Koza. Finally I would like to thank all of my committee members for supervising my work with continuous encouragement: Dr. William Crowe, Dr. Robert Strongin, Dr. Steven Watkins and Dr. Ding Shih. iii TABLE OF CONTENTS DEDICATION ...................................................................................................ii ACKNOWLEDGEMENTS .............................................................................iii LIST OF TABLES.............................................................................................vi LIST OF FIGURES ...........................................................................................vii LIST OF ABBREVIATIONS ...........................................................................xi ABSTRACT........................................................................................................xv CHAPTER 1. ACID LABILE LINKERS FOR SOLID PHASE SYNTHESIS OF PEPTIDES....................................................1 1.1 Introduction........................................................................................1 1.2 Literature Review...............................................................................7 CHAPTER 2. DESIGN AND SYNTHESIS OF NOVEL FERROCENE LINKER………………………………………………………36 2.1 Introduction........................................................................................36 2.2 Design of the Ferrocene Linker..........................................................38 2.3 Results and Discussion ......................................................................39 2.4 Experimental......................................................................................45 2.4.1 4-Ferrocenyl-4-oxobutanoic Acid.............................................45 2.4.2 2-Propenyl-4-ferrocenyl-4-oxobutanoate..................................46 2.4.3 Ferrocenyl-1-hydrazide butanoic Acid......................................47 2.4.4 2-Methyl 4-ferrocenyl-4-oxohexanoate ....................................47 2.4.5 Ferrocenyl-1 hydrazide hexanoic Acid .....................................48 2.4.6 Ferrocenyl-1,1'-acid Fluoride....................................................48 2.4.7 Methyl ferrocenecarboxylate.....................................................49 2.4.8 Methyl-1'-formyl-1-ferrocene carboxylateferrocene.................50 2.4.9 Hept-6-enylamine......................................................................50 2.4.10 Ferrocenyl-imine-glycine(ethylester), methoxycarbonyl ........51 2.4.11 Ferrocene-methylglycine-ethyl ester, methoxycarbonyl .........52 2.4.12 Ferrocene-1-carboxylic acid-1`carboxaldehyde......................52 2.4.13 Ferrocene carboxaldehyde Resin ............................................53 2.4.14 Ferrocenyl heptenoic amine Resin..........................................57 2.4.15 Agni[(KLAKKLA)2] (2.24)using ferrocenyl heptenoic amine Resin............................................................................57 2.4.16 Ferrocene carboxaldehyde-polystyrene Resin.........................58 2.4.17 Allyl-amino ferrocenyl polystyrene Resin ..............................59 2.4.18 Allyl-amino fmoc-phenylalanine ferrocenyl polystyrene Resin .......................................................................................59 iv 2.4.19 Cleavage of (1-allylcarbamoyl-2-phenyl-ethyl)-carbamic acid 9H-fluoren-9-ylmethyl ester from allyl-amino ferrocenyl polystyrene Resin...................................................60 CHAPTER 3. CONVERGENT PEPTIDE SYNTHESIS ............................61 3.1 Introduction........................................................................................61 3.2 Secondary Structure ...........................................................................64 3.3 Solid-Phase Peptide Synthesis Methods ............................................66 3.4 Convergent Peptide Synthesis............................................................70 3.4.1 Convergent Solid-Phase Peptide Synthesis(CSPPS) of Protected Peptide Segments......................................................71 3.4.2 Chemoselective ligation of Unprotected Peptide Fragments..................................................................................74 3.5 Results and Discussion ......................................................................77 3.6 Experimental......................................................................................81 3.6.1 Peptide Synthesis ......................................................................81 3.6.2 Cyh-7.........................................................................................84 3.6.3 Cyh-7 MIC Assays....................................................................85 3.6.4 16-Hydroxypalmitoleic Acid ....................................................86 3.6.5 Peptide Synthesis Agni[ Glu(O-All)(KLAKKLA)2heptenoic Acid] .....................87 3.7 Ring Closing Methatesis Peptide Ligation............................................87 3.7.1 Attempted Peptide Ligation ......................................................88 3.7.2 Attempted Peptide Ligation ......................................................88 CHAPTER 4 CONCLUSION AND FUTURE STUDIES ..........................89 4.1 Discussion.............................................................................................89 REFERENCES.............................................................................................. …90 VITA....................................................................................................................96 v LIST OF TABLES Table 1.1 Common protecting groups used in SPPS where side chains are cleaved mild to moderate acidic conditions........................................5 Table 3.1 Common coupling reagents used in SPPS .........................................69 Table 3.2 Examples of large synthetic peptides and small proteins synthesized by SPPS...........................................................................70 Table 3.3 Methods for ligating unprotected peptides..........................................75 Table 3.4 Minimum inhibitory concentration of Cyh-7 ......................................86 vi LIST OF FIGURES Figure 1.1 Schematic representation of a peptide synthesis...............................1 Figure 1.2 Purification of compounds bound to the solid support from those in solution by simple filtration............................2 Figure 1.3 General scheme for solid-phase peptide synthesis page...................4 Figure 1.4 Cleavage of peptide from the linker PAL.........................................7 Figure 1.5 Cleavage of peptide from BAL(Backbone amide linker) .................8 Figure 1.6 Cleavage of peptide from trityl resin ................................................9 Figure 1.7 Cleavage of peptide from the Rink linker.........................................10 Figure 1.8 Methoxy substituted benzyl amine linkers ......................................11 Figure 1.9 Peptide cleavage from BDMTA resin .............................................11 Figure 1.10 Cleavage of peptide from BHA resin ..............................................12 Figure 1.11 Cleavage of peptide from MBHA resin...........................................13 Figure 1.12 Peptide cleavage using Alkoxybenzylamine linker