Protease-mediated fragmentation of p-amidobenzyl ethers: a new strategy for the activation of anticancer prodrugs

J Org Chem. 2002 Mar 22;67(6):1866-72. doi: 10.1021/jo016187+.

Abstract

A new anticancer prodrug activation strategy based on the 1,6-elimination reaction of p-aminobenzyl ethers is described. Model studies were undertaken with the N-protected peptide benzyloxycarbonyl-valine-citrulline (Z-val-cit), which was attached to the amino groups of p-aminobenzyl ether derivatives of 1-naphthol and N-acetylnorephedrine. The amide bond that formed was designed for hydrolysis by cathepsin B, a protease associated with rapidly growing and metastatic carcinomas. Upon treatment with the enzyme, the Z-val-cit-p-amidobenzyl ether of 1-naphthol (2) underwent peptide bond hydrolysis with the rapid release of 1-naphthol. The aliphatic Z-val-cit-p-amidobenzyl ether of N-acetylnorephedrine (5) also underwent amide bond hydrolysis, but without the ensuing elimination of N-acetylnorephedrine. On the basis of these results, the phenolic anticancer drugs etoposide (6) and combretastatin A-4 (7) were attached to the Z-val-cit-p-amidobenzyl alcohol through ether linkages, forming the peptide-drug derivatives 8 and 9, respectively. Both compounds were stable in aqueous buffers and serum and underwent ether fragmentation upon treatment with cathepsin B, resulting in the release of the parent drugs in chemically unmodified forms. The released drugs were 13-50 times more potent than were the prodrug precursors on a panel of cancer cell lines. In contrast, the corresponding carbonate derivative of combretastatin A-4 (13) was unstable in aqueous environments and was as cytotoxic as combretastatin A-4. This result extends the use of the self-immolative p-aminobenzyl group for the fragmentation of aromatic ethers and provides a new strategy for anticancer prodrug development.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antineoplastic Agents
  • Benzyl Alcohols / chemistry*
  • Catalysis
  • Cathepsin B / metabolism
  • Chromatography, High Pressure Liquid
  • Dipeptides / chemistry*
  • Drug Screening Assays, Antitumor
  • Endopeptidases / metabolism*
  • Humans
  • Hydrolysis
  • Lung Neoplasms
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Structure
  • Naphthols / chemistry
  • Phenyl Ethers / chemistry*
  • Prodrugs / chemistry*
  • Tumor Cells, Cultured / drug effects

Substances

  • Antineoplastic Agents
  • Benzyl Alcohols
  • Dipeptides
  • Naphthols
  • Phenyl Ethers
  • Prodrugs
  • Z-valine-citrulline-p-aminobenzylalcohol
  • 1-naphthol
  • Endopeptidases
  • Cathepsin B