Tight-binding inhibitors-IV. Inhibition of adenosine deaminases by various inhibitors

Ram P. Agarwal, Thomas Spector, Robert E. Parks

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359 Scopus citations


Three ADA (adenosine deaminase) inhibitors, DHMPR (1,6-dihydro-6-hydroxymethyl purine ribonucleoside); EHNA [erythro-9-(2-hydroxy-3 nonyl)adenine] ; and deoxycoformycin [(R)-3-(2-deoxy-β-d-erythro-pento-furanosyl)-3, 6,7,8-tetrahydroimidazo[4,5-d] [1,3-diazepin-8-ol] or Covidarabin, were compared with regard to their inhibitory behavior with ADAs from human erythrocytes and calf intestine. Marked differences in the times required for establishment of steady state between the enzyme and inhibitors were observed, e.g. DHMPR, virtually instantaneous; EHNA, 2-3 min; and deoxycoformycin, many hr. The parameters of the inhibition of human erythrocytic ADA by deoxycoformycin were as follows: the association rate constant (k1) = 2.6 × 106 M-1 sec-1 ; the dissociation rate constant of the enzyme-inhibitor complex (k2) = 6.6 × 10-6 sec-1; Ki (from k2 k1) = 2.5 × 10-12M and Ki (from I50) = 1.5 × 10-11 M. The Ki values for EHNA and DHMPR, as determined by classical methods after attainment of steady state, were 1.6 × 10-9 and 1.3 × 10-6 M, respectively, for human erythrocytic ADA. The kinetic parameters for EHNA and calf intestinal ADA were as follows: Ki = 6.5 × 10-9 M (by the method of I50); k1 = 0.7 × 106 M-1 sec-1' and k2 = 4.6 × 10-3 sec-1. On the basis of Ki values, the inhibitors. DHMPR, EHNA and deoxycoformycin (a transition state analog), were classified as readily reversible, semi-tight-binding and tight-binding inhibitors. The difficulties encountered in the kinetic analyses of different types of inhibitors and the methods for dealing with the problems of these inhibitors are discussed.

Original languageEnglish (US)
Pages (from-to)359-367
Number of pages9
JournalBiochemical Pharmacology
Issue number5
StatePublished - Mar 1 1977

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology


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