Examining the Chirality, Conformation and Selective Kinase Inhibition of 3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (CP-690,550)

Abstract

Here, we examine the significance that stereochemistry plays within the clinically relevant Janus Kinase 3 (Jak3) inhibitor CP-690,550. A synthesis of all four enantiopure stereoisomers of the drug was carried out and an examination of each compound revealed that only the enantiopure 3R, 4R isomer was capable of blocking Stat5 phosphorylation (Jak3 dependent). Each compound was profiled across a panel of over 350 kinases which revealed a high level of selectivity for the Jak family kinases for these related compounds. Each stereoisomer retained a degree of binding to Jak3 and Jak2 and the 3R, 4S and 3S, 4R stereoisomers were further revealed to have binding affinity for selected members of the STE7 and STE20 subfamily of kinases. Finally, an appraisal of the minimum energy conformation of each stereoisomer and molecular docking at Jak3 was performed in an effort to better understand each compounds selectivity and potency profiles.

Selective functional inhibition of JAK‐3 is sufficient for efficacy in collagen‐induced arthritis in mice

“…Despite excellent selectivity across the kinome in general, CP-690,550 was recently reported to have modest selectivity against JAK-1 (4-fold) and JAK-2 (3-fold) and considerable selectivity against Tyk-2 (357-fold) (16), which is consistent with our cell-free assays (5-fold versus JAK-1, 10-fold versus JAK-2, 20-fold versus Tyk-2). This is not unexpected, given that a crystallographic model of CP-690,550 at the active site of the JAK-3 kinase postulates that only 3 residues proximal to the inhibitor differ across the JAK family, and these differences are conservative mutations (16,37).…”

“…This is not unexpected, given that a crystallographic model of CP-690,550 at the active site of the JAK-3 kinase postulates that only 3 residues proximal to the inhibitor differ across the JAK family, and these differences are conservative mutations (16,37). Nevertheless, based on precedent from JAK-3-deficient SCID patients and the potential limitations of inhibiting JAK-1, we have chosen to optimize for in vivo JAK-3 selectivity and have benchmarked the potential utility of WYE-151650 against that of CP-690,550 for measures of both in vivo efficacy and selectivity.…”

“…The structures of JAK-3 inhibitors (WYE-151650, WYE-152038, and CP-690,550) are shown in Figure 1A, and their synthesis was described previously (16,17). Compounds were dissolved in vehicle containing 0.5% methylcellulose and 0.25% Tween for in vivo studies, unless otherwise indicated.…”

“…Although CP-690,550 inhibits JAK-3, it also has an overlapping activity against JAK-1 and JAK-2 (15,16). This fact raises the question of whether inhibition of JAK-3 alone is sufficient to disrupt cytokine signaling and to result in amelioration of autoimmune disease pathology.…”

Towards Selective Inhibitors of Janus Kinase 3: Identification of a Novel Structural Variation between Janus Kinases 2 and 3

“…Several synthetic JAK3 inhibitors identified by pharmaceutical companies are currently in the process of clinical evaluation. In particular, CP-690,550, a highly potent inhibitor against JAK3 (IC 50 = 1 nM), showed significant extension of life of the organ-transplanted animal 8,9 and demonstrated efficacy in phase III clinical trials for the treatment of rheumatoid arthritis 10,11 and rejection in kidney transplant patients.…”