Minimization of Parathyroid Hormone Using Simultaneous Multiple Peptide Synthesis: Implications for Structure Based Drug Design

  • Ashok Khatri
  • Xiang-Chen Huang
  • Brian D. Petroni
  • Thomas J. Gardella
Part of the American Peptide Symposia book series (APSY, volume 7)

Abstract

Parathyroid hormone (PTH), an 84 residue long polypeptide, acts on PTH-1 receptors in bone and kidney to control blood calcium levels. Synthetic PTH(l-34), one of the first peptide hormones to be chemically synthesized, has full biological activity [1] (EC50 for cAMP generation ≈2 nM). Based on previous peptide truncation studies that identified residues 15–31 as the principal receptor-binding domain and residues 1–14 as the principal signaling domain, PTH(1–14) was synthesized and found to have faint but measurable activity (EC50 ≈200 μM) [2]. The peptide was prepared on a multiple peptide synthesizer and the crude peptide was found to be of high quality, as shown by HPLC and MALDI-MS, and could be used directly in bioactivity assays after a simple desalting step on a disposable C18 cartridge. This finding prompted us to use this approach to synthesize an extensive series of PTH(1–14) analogs with which we could define structure-activity relationships in the peptide and potentially improve potency. Within a short time (≈1.5 years) we generated and characterized over 200 PTH(1–14) analogs and have thus far improved potency at least 1,000 times.

Keywords

HPLC Osteoporosis Polypeptide Glucagon Calcitonin 

References

  1. 1.
    Tregear, G.W., Van Rietschoten, J., Greene, E., et al. Endocrinology 93, 1349–1353 (1973).PubMedCrossRefGoogle Scholar
  2. 2.
    Luck, M., Carter, P., Gardella T. Mol. Endocrinol. 13, 670–680 (1999).PubMedGoogle Scholar
  3. 3.
    Shimizu, M., Potts, J.T., Jr., Gardella, T. J. Biol. Chem. 275, 21836–21843 (2000).PubMedCrossRefGoogle Scholar
  4. 4.
    Shimizu, Carter, P., Khatri, A., et al. Endocrinology 142, 3068–3074 (2001).PubMedGoogle Scholar
  5. 5.
    Carter, P., Gardella, T. Biochim. Biophys. Acta 1538, 290–304 (2001).PubMedCrossRefGoogle Scholar
  6. 6.
    Neer, R., Arnaud, C., Zanchetta, J., et al. N.E.J.M. 344, 1434–1441 (2001).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2001

Authors and Affiliations

  • Ashok Khatri
    • 1
    • 2
  • Xiang-Chen Huang
    • 1
  • Brian D. Petroni
    • 1
  • Thomas J. Gardella
    • 1
    • 2
  1. 1.Massachusetts General HospitalUSA
  2. 2.Harvard Medical SchoolBostonUSA

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