University of New Hampshire University of New Hampshire Scholars' Repository Doctoral Dissertations Student Scholarship Spring 1975 PART I THE SYNTHESIS OF PROLINE-CONTAINING POLYPEPTIDES PART II CHEMICAL SHIFT NONEQUIVALENCE IN 2,4-DINITROPHENYL SULFENAMIDES THOMAS DAVID HARRIS Follow this and additional works at: https://scholars.unh.edu/dissertation Recommended Citation HARRIS, THOMAS DAVID, "PART I THE SYNTHESIS OF PROLINE-CONTAINING POLYPEPTIDES PART II CHEMICAL SHIFT NONEQUIVALENCE IN 2,4-DINITROPHENYL SULFENAMIDES" (1975). Doctoral Dissertations. 1082. https://scholars.unh.edu/dissertation/1082 This Dissertation is brought to you for free and open access by the Student Scholarship at University of New Hampshire Scholars' Repository. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of University of New Hampshire Scholars' Repository. For more information, please contact [email protected]. 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Xerox University Microfilms 300 North Zeeb Road Ann Arbor, Michigan 48106 75-19,874 HARRIS, Thomas David, 1948- PART I. THE SYNTHESIS OF PROLINE-CONTAINING POLYPEPTIDES. PART II. CHEMICAL SHIFT NONEQUIVALENCE IN 2,4 -DINITROPHENYLSULFENAMIDES. University of New Hampshire, Ph.D., 1975 Chemistry, organic Xerox University Microfilms, Ann Arbor, Michigan 48106 THIS DISSERTATION HAS BEEN MICROFILMED EXACTLY AS RECEIVED. PART I. THE SYNTHESIS OF PROLINE-CONTAINING POLYPEPTIDES PART II. CHEMICAL SHIFT NONEQUIVALENCE IN 2,4-DINITROPHENYLSULFENAMIDES by THOMAS D. HARRIS B. S., Saginaw Valley College, 1971 A THESIS Submitted to the University of New Hampshire In Partial Fulfillment of The Requirements for the Degree of Doctor of Philosophy Graduate School Department of Chemistry May, 1975 This thesis has been examined and approved. /I Thesis director, Robert E. Lyle Professor of Chemistry Jame5s5 DD.. MMoorrrriissoonn Professor of Chemistry 3. John Uebel, Professor of Chemistry N. Dennis Chasteen, Associate Professor of Chemistry Gerald L. Klippenstein Associate Professor of Biochemistry Date thesis is dedicated to my wife, Susan, and to my parents. ACKNOWLEDGEMENTS The author would like to express his appreciation to Dr. Robert E. Lyle for providing and directing the research problem and for his guidance and enthusiasm during all stages of the research. He would also like to thank Dr. James D. Morrison and Dr. J. John Uebel for their helpful discussions during the course of the research and for their aid in the development of this thesis manuscript. To the other members of the Faculty and Staff of the Chemistry Department, the author wishes to express his apprec­ iation for the educational opportunity which they have pro­ vided. Special thanks are also given to Mrs. Deanna Cardin for performing the elemental analyses and to Miss Anne Kohl for her assistance in the preparation of this manuscript. Thanks are extended also to Rita Furman for the anti­ biotic assay and to the National Institutes of Health for providing the results of the antifertility screening. The author expresses his appreciation to the University of New Hampshire for providing him with financial support in the way of UNH Summer Fellowships. Finally, the author would like to thank his colleague Dr. Richard Davenport for his consultation and encouragement during the course of this research project. TABLE OF CONTENTS LIST OF TABLES............................................... xiii LIST OF FIGURES............................................... xiv ABSTRACT........................................................ xv PART I. THE SYNTHESIS OF PROLINE-CONTAINING POLYPEPTIDES1 SECTION I. INTRODUCTION....................................... 2 SECTION II. RESUSTS AND DISCUSSION......................... 10 General Considerations of Peptide Synthesis............ 10 Preparation of N-Hexanoylglycyl-L-prolyl-5- aminovaleric Acid (19j............................. 17 An Alternate Approach to N-Hexanoylglycyl-L- prolyl-5-arainovaleric Acid (lj))................... 23 Preparation of N-Pentanoylglycyl-L-prolyl-5- aminovaleric Acid Methyl Ester (3C0.............. 2 8 Preparation of N-Hexanoylglycyl-L-prolylglycyl- glycine (^6).........................................31 Synthesis of a FSH/LH-RF Analog ........................ 34 Preparation of N-Hexanoylglycly-L-prolyl-L-leucyl- L-leucine (56j...................................... 35 Preparation of N-Pentanoylglycyl-L-prolyl-L- luecyl-L-leucine (159).............................. 46 Preparation of N-Pentanoylglycyl-D-prolyl-L- phenylalanyl-L-leucine {19).........................48 Attempted Preparation of Derivatives of 5- Hydroxyproline...................................... 54 Biological Results ....................................... 60 v SECTION III. EXPERIMENTAL.................................... 64 General.................................................... 64 Melting Points...................................... 64 Boiling Points...................................... 64 Elemental Analyses.................................. 64 Infrared Spectra.................................... 64 Nuclear Magnetic Resonance Spectra................. 64 Materials and Methods.............................. 65 Reagents....................................... 65 Solvents........................................66 Products........................................66 Experimental Methods.......................... 66 Thin Layer Chromatography.................... 67 Preparation of Hexanoyl Chloride (10^).................. 6 8 Preparation of N-Hexanoylglycine Ethyl Ester (15) .... 68 Preparation of N-Hexanoylglycine (!L2).................. 68 Method A ............................................. 68 Method B ............................................. 69 Method C ............................................. 69 Preparation of N-Hexanoylglycyl-L-prolyl-5-amino- valeric Acid (19J . Method A ..................... 70 Attempted Preparation of N-Hexanoylglycyl-L-prolyl- 5-aminovaleric Acid (19J........................... 72 Preparation of N-Benzyloxycarbonyl-L-proline (2_2) ... .73 Preparation of Methyl 5-Aminovalerate Hydrochloride (21).................................................. 74 Preparation of L-Prolyl-5-aminovaleric Acid Methyl Ester (25)................................... 74 Preparation of N-Hexanoylglycyl-L-prolyl-5-amino- valeric Acid Methyl Ester (21).................... 76 Preparation of N-Hexanoylglycyl-L-prolyl-5-amino- Acid (19^). Method B ............................... 77 Preparation of Pentanoyl Chloride (28)................ 77 Preparation of N-Pentanoylglycine (29)................ 77 Method A ............................................. 77 Method B ............................................. 78 vi Preparation of N-Pentanoylglycyl-L-prolyl-5-amino- valeric Acid Methyl Ester (30^).................. 78 Method A .............................................78 Method B .......................................79 Preparation of L-Proline Methyl Ester (_31).............80 Preparation of N-Hexanoylglycyl-L-proline Methyl Ester (32^........................................... 80 Preparation of N-Hexanoylglycyl-L-proline (33)....... 81 Method A ............................................. 81 Method B ............................................. 82 Preparation of N-Hexanoylglycyl-L-prolylglycyl- glycine Benzyl Ester (350......................... 82 Method A ............................................. 82 Method B ............................................. 83 Preparation of N-Hexanoylglycyl-L-prolylglycyl- glycine (36)........................................ 83 Preparation of N-Hexanoylglycyl-L-prolylglycyl- glycinamide