Michael W. Pennington

Peptide synthesis protocols

Procedures to Improve Difficult Couplings, Michael W. Pennington and Michael E. Byrnes. Methods for Removing the Fmoc Group, Gregg B. Fields. Solvents for Solid-Phase Peptide Synthesis, Cynthia G. Fields and Gregg B. Fields. HF Cleavage and Deprotection Procedures for Peptides Synthesized Using a Boc/Bzl Strategy, Michael W. Pennington. Acid Cleavage/Deprotection in Fmoc/tBu-Solid Phase Peptide Synthesis, Fritz Dick. Bromoacetylated Synthetic Peptides: Starting Materials for Cyclic Peptides, Peptomer, and Peptide Conjugates. Frank A. Robey. Formation of Disulfide Bonds in Synthetic Peptides and Proteins, David Andreau, Fernando Albericio, Nuria A. Sole, Mark C. Munson, Marc Ferrer, and George Barany. Site-Specific Chemical Modification Procedures, Michael W. Pennington. Synthesis of Phosphopeptides Containing O-Phosphoserine and O-Phosphothreonine, Anatol Arendt and Paul A. Hargrave. Solid-Phase Synthesis of Phosphorylated Tyr-Peptides by Phosphite Triester Phosphorylation, Michael W. Pennington. Design of Novel Synthetic Peptides Including Cyclic Conformationally and Topographically Constrained Analogs, Victor J. Hruby and G. Gregg Bonner. Solid-Phase Synthesis of Peptides Containing the CH2NH Reduced Bond Surrogate, Michael W. Pennington. Approaches to the Asymmetric Synthesis of Unusual Amino Acids, Victor J. Hruby and Xinhua Qian. Synthesis of Fully Protected Peptide Fragments, Monika Mergler. Peptide Synthesis via Fragment Condensation, Rolf Nyfeler. Index.

Peptide analysis protocols

Gel-Filtration Chromatography Daniel M. Bollag. Ion-Exchange Chromatography Daniel M. Bollag. Reversed-Phase HPLC: Analytical Procedure Udo Nirenberg. Reversed-Phase High-Performance Liquid Chromatography: A Semipreparative Methodology Michael E. Byrnes. Applications of Strong Cation-Exchange (SCX)-HPLC in Synthetic Peptide Analysis Dan L. Crimmins. Principles and Practice of Peptide Analysis with Capillary Zone Electrophoresis Thomas E. Wheat. Fast Atom Bombardment Mass Spectrometric Characterization of Peptides P. R. Das and B. N. Pramanik. Sequence Analysis of Peptide Resins from Boc/Benzyl Solid- Phase Synthesis Jan Pohl. NMR Spectroscopy of Peptides and Proteins Mark G. Hinds and Raymond S. Norton. Techniques for Conjugation of Synthetic Peptides to Carrier Molecules J. Mark Carter. Epitope Prediction Methods J. Mark Carter. Epitope Mapping of a Protein Using the Geysen (PEPSCAN) Procedure J. Mark Carter. Analysis of Proteinase Specificity by Studies of Peptide Substrates:The Use of UV and Fluorescence Spectroscopy to Quantitate Rates of Enzymatic Cleavage Ben M. Dunn Paula E. Scarborough Ruth Davenport and Wieslaw Swietnicki. Synthesis of Recombinant Peptides Gino Van Heeke Jay S. Stout and Fred W. Wagner. De Novo Design of Proteins: Template-Assembled Synthetic Proteins (TASP) Gabriele Tuchscherer Verena Steiner Karl- Heinz Altmann and Manfred Mutter. Chemical Synthesis of the Aspartic Proteinase from Human Immunodeficiency Virus (HIV) Paul D. Hoeprich Jr. Multiple and Combinatorial Peptide Synthesis: Chemical Development and Biological Applications Philip C. Andrews Daniele M. Leonard Wayne L. Cody and Tomi K. Sawyer. Index.

CHAPTER 10:Case Study 2: Transforming a Toxin into a Therapeutic: the Sea Anemone Potassium Channel Blocker ShK Toxin for Treatment of Autoimmune Diseases

Effector memory T lymphocytes, which are involved in autoimmune diseases such as multiple sclerosis, type 1 diabetes mellitus and rheumatoid arthritis, express Kv1.3 potassium channels, which play a major role in their activation. Blockers of lymphocyte Kv1.3 channels preferentially inhibit the activation of these cells and therefore show considerable potential as therapeutics for autoimmune diseases. ShK, a 35-residue polypeptide isolated from the Caribbean sea anemone Stichodactyla helianthus, blocks Kv1.3 channels at picomolar concentrations. Although ShK was effective in treating rats with delayed type hypersensitivity and a model of multiple sclerosis, it lacked selectivity for Kv1.3 channels over closely related Kv1 channels. Extensive mutagenesis studies combined with elucidation of the structure of ShK provided the basis for developing new ShK analogues with improved selectivity and increasing stability, which have proven efficacious in preventing and/or treating animal models of delayed type hypersensitivity, type 1 diabetes, rheumatoid arthritis, and multiple sclerosis without inducing generalized immunosuppression. They are currently undergoing clinical evaluation as potential immunomodulators for the treatment of autoimmune diseases.

Kv1.3 Selective Peptides Based upon N-Terminal Extension and Internal Substitutions of ShK Toxin

With more than 80 different types of autoimmune disorders affecting all organ systems in the human body, finding a drug which would treat patients at the root cause of these diseases has been a quest of our mutual labs for the past two decades. These diseases may range from mild skin disorders such as psoriasis, widely distributed joint damage such as in rheumatoid arthritis, destruction of specific cells such as β cells in type-1 diabetes to more complex disorders affecting central nervous system such as multiple sclerosis. Current treatments involve the use of broad immunosuppressants, which may open the door to opportunistic infections. Through work pioneered with the Chandy lab, we have shown that the channel phenotypes of these autoantigen specific T-cells have preferentially upregulated the Kv1.3 channel to balance Ca influx upon activation. A sea anemone derived peptide named ShK is one of the most potent Kv1.3 blockers described (IC50 = 10 pM), however it lacks specificity and blocks Kv1.1 and Kv1.5 also at pM levels [1,2]. Through many years of engineering, we have progressed one of our peptides, Dalazatide (formerly ShK-186) to the clinic for its selective block of Kv1.3 channels as a means of treating autoimmune diseases. In our current work, we have built upon our findings to continue to improve the selectivity of ShK-derived analogs and have recently reported selectivity profiles of more than 1000x for Kv1.3 versus Kv1.1. We have improved the drugability of ShK [3,4].

Role of disulfide bonds in the structure and activity of ShK toxin

Michael W. Pennington, Mark Lanigan, Vladimir M. Mahnir, Kati Kalman, Cheryl T. McVaugh, David Behm, Denise Donaldson, K. George Chandy, William R. Kem, and Raymond S. Norton2 Bachem Bioscience Inc., King of Prussia, PA 19406, U.S.A.; Biomolecular Research Institute, Parkville 3052 Australia; Department of Pharmacology and Therapeutics, College of Medicine, University of Florida, Gainesville, FL 32610-0267, U.S.A.; and Dept. of Physiology and Biophysics, University of California, Irvine, CA 92697, U.S.A.