Diadenosine 5',5''-(boranated)polyphosphonate analogues as selective nucleotide pyrophosphatase/phosphodiesterase inhibitors

J Med Chem. 2010 Dec 23;53(24):8485-97. doi: 10.1021/jm100597c. Epub 2010 Nov 22.

Abstract

Nucleotide pyrophosphatase/phosphodiesterases (NPPs) hydrolyze extracellular nucleotides and dinucleotides and thus control purinergic signaling. Enhanced NPP activity is implicated in health disorders such as osteoarthritis and cancer. We designed novel diadenosine polyphosphonate derivatives as potential NPP inhibitors. Analogues 1-4 bear a phosphonate and/or boranophosphate group and/or a 2'-H atom instead of a 2'-OH group. In comparison to ATP, analogues 1-4 were barely hydrolyzed by human NTPDase1, -2, -3, and -8 (<5% hydrolysis) and NPP1 and -3 (≤ 13%) and were not hydrolyzed by ecto-5'-nucleotidase, unlike AMP. These derivatives did not affect NTPDase activity, and analogues 1 and 2 did not inhibit ecto-5'-nucleotidase. All analogues blocked ∼80% of the NPP2-dependent hydrolysis of pnp-TMP, a specific NPP substrate, and inhibited the catabolism of pnp-TMP (K(i) and IC₅₀ both found to be between 10 and 60 μM), Ap₅A, and ATP by NPP1. The activity of NPP3 was inhibited to a lesser extent by the new analogues, with compounds 1 and 4 being the most effective in that respect. The analogues dramatically reduced the level of hydrolysis of pnp-TMP at the cell surface of both osteocarcinoma and colon cancer cells. Importantly, analogues 1-4 exhibited significantly reduced agonistic activity toward human P2Y₁,₁₁) receptors (except for analogue 1) and no activity with human P2Y₂ receptor. Our data provide strong evidence that analogue 2 is the first specific NPP inhibitor to be described.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine Nucleotides / chemical synthesis*
  • Adenine Nucleotides / chemistry
  • Adenine Nucleotides / pharmacology
  • Adenosine Triphosphate / metabolism
  • Boranes / chemical synthesis*
  • Boranes / chemistry
  • Boranes / pharmacology
  • Calcium / metabolism
  • Cell Line
  • Dinucleoside Phosphates / chemical synthesis*
  • Dinucleoside Phosphates / chemistry
  • Dinucleoside Phosphates / metabolism
  • Dinucleoside Phosphates / pharmacology
  • Drug Screening Assays, Antitumor
  • Humans
  • Hydrolysis
  • Nucleotidases / metabolism
  • Phosphoric Diester Hydrolases
  • Purinergic P2Y Receptor Agonists / chemical synthesis
  • Purinergic P2Y Receptor Agonists / chemistry
  • Purinergic P2Y Receptor Agonists / pharmacology
  • Pyrophosphatases / antagonists & inhibitors*
  • Receptors, Purinergic P2Y / physiology
  • Structure-Activity Relationship
  • Substrate Specificity
  • Thymidine Monophosphate / analogs & derivatives
  • Thymidine Monophosphate / metabolism

Substances

  • Adenine Nucleotides
  • Boranes
  • Dinucleoside Phosphates
  • Purinergic P2Y Receptor Agonists
  • Receptors, Purinergic P2Y
  • thymidine 5'-4-nitrophenyl phosphate
  • Thymidine Monophosphate
  • P(1),P(5)-di(adenosine-5'-)pentaphosphate
  • Adenosine Triphosphate
  • Nucleotidases
  • Phosphoric Diester Hydrolases
  • Pyrophosphatases
  • nucleotide pyrophosphatase - phosphodiesterase I
  • Calcium