Abstract
We present the in vitro characterization of a novel phosphodiesterase type 4 inhibitor, CDP840 (R-[+]-4-[2-{3-cyclopentyloxy-4-methoxyphenyl}-2-phenylethyl]pyridine), which has shown efficacy in a phase II allergen challenge study in asthmatics without adverse effects. CDP840 potently inhibits PDE-4 isoenzymes (IC50 2–30 nM) without any effect on PDE-1, 2, 3, 5, and 7 (IC50>100 μM). It exhibited no significant selectivity in inhibiting human recombinant isoenzymes PDE-4A, B, C or D and was equally active against the isoenzymes lacking UCR1 (PDE-4B2 and PDE-4D2). In contrast to rolipram, CDP840 acted as a simple competitive inhibitor of all PDE-4 isoenzymes. Studies with rolipram indicated a heterogeneity within all the preparations of PDE-4 isoenzymes, indicative of rolipram inhibiting the catalytic activity of PDE-4 with both a low or high affinity. These observations were confirmed by the use of a PDE-4A variant, PDE-4A330–886, which rolipram inhibited with low affinity (IC50=1022 nM). CDP840 in contrast inhibited this PDE-4A variant with similar potency (IC50=3.9 nM), which was in good agreement with theK d of 4.8 nM obtained from [3H]-CDP840 binding studies. Both CDP840 and rolipram inhibited the high-affinity binding of [3H]-rolipram binding to PDE-4A, B, C and D with similarK d app (7–19 nM and 3–5 nM, respectively). Thus, the activity of CDP840 at the [3H]-rolipram binding site was in agreement with the inhibitor’s activity at the catalytic site. However, rolipram was ∼100-fold more potent than CDP840 at inhibiting the binding of [3H]-rolipram to mouse brain in vivo. These data clearly demonstrate that CDP840 is a potent selective inhibitor of all PDE-4 isoenzymes. In contrast to rolipram, CDP840 was well-tolerated in humans. This difference, however, cannot at present be attributed to either isoenzyme selectivity or lack of activity in vitro at the high-affinity rolipram binding site (Sr).
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Abbreviations
- PDE:
-
phosphodiesterase
- cAMP:
-
adenosine 3′−5′-cyclic monophosphate
- BSA:
-
bovine serum albumin
- Sr:
-
high-affinity rolipram binding site
- Sc:
-
low-affinity rolipram binding site
- AEBSF:
-
4-(-2-aminoethyl) benzenesulfonyl fluoride
- EDTA:
-
ethylenediaminetetraacetic acid
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