Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.790289
Title: Reverse native chemical ligation as a new entry to cyclic peptide therapeutics
Author: Shariff, L. S.
ISNI:       0000 0004 8503 978X
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2016
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Abstract:
Cyclic peptides are of particular therapeutic interest, often exhibiting improved biological activity and resistance to in vivo proteolysis over their linear counterparts, but current production methods can be costly, lengthy and complex. The adaptation of a facile way to join peptide fragments developed by the Macmillan group forms the basis of the investigation and application of a novel method of peptide cyclisation featured within this thesis. A one-pot, tandem reaction of reverse native chemical ligation (retro-NCL via N→S acyl shift) at a C-terminal cysteine, in the presence of a thiol additive to 'trap' the generated transient thioester, followed by NCL (S→N) at an N-terminal cysteine on the same peptide chain resulted in the successful cyclisation of various biologically active sequences. Each circular peptide underwent testing for novel therapeutic roles, including a sunflower trypsin inhibitor analogue SFTI(I10H) that was found, upon design of an enzymatic assay, to inhibit human kallikrein 5 (implicated in atopy and epithelial cancers) with IC50 0·14 µM. To facilitate studies on the influence of pKas on reaction rate, synthesis routes to the production of orthogonally protected β-cysteine and γ-cysteine for use in standard solid-phase peptide synthesis were established. In combination with studies using pH variation it was found that the pKa of the amino group participating in the N→S acyl shift does not completely explain variations in reaction rate observed for C-terminal cysteine carboxylates versus carboxamides. Instead it is proposed that the rate of the acyl transfer during retro-NCL is related to stability induced by zwitterion formation for carboxylates but α-amino group pKa values for carboxamides. The importance of the thiol additive identity was also highlighted, with the example captamine being prone to thioester formation in addition to cyclisation under the conditions used.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.790289  DOI: Not available
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