Testing the Promiscuity of Commercial Kinase Inhibitors Against the AGC Kinase Group Using a Split-luciferase Screen

Abstract

Using a newly developed competitive binding assay dependent upon the reassembly of a split reporter protein, we have tested the promiscuity of a panel of reported kinase inhibitors against the AGC group. Many non-AGC targeted kinase inhibitors target multiple members of the AGC group. In general, structurally similar inhibitors consistently exhibited activity toward the same target as well as toward closely related kinases. The inhibition data was analyzed to test the predictive value of either using identity scores derived from residues within 6 Å of the active site or identity scores derived from the entire kinase domain. The results suggest that the active site identity in certain cases may be a stronger predictor of inhibitor promiscuity. The overall results provide general guidelines for establishing inhibitor selectivity, as well as for the future design of inhibitors that either target or avoid AGC kinases.

Irreversible Inhibitors of c-Src Kinase That Target a Nonconserved Cysteine

“…Profiling of 131 kinases was performed at 15 nM (5x IC 50 in this assay) dasatinib or compound 9 using a 2 h incubation (Luceome Biotechnologies). 24,25 Selectivity differences between compounds can be compared by their S -score. 26 S -score represents the fraction of kinases inhibited to a particular level ( S 35 is the number of kinases with <35% of control activity divided by the total number of non-mutant kinases in the panel, here 124).…”

TGFβ Inhibition Stimulates Collagen Maturation to Enhance Bone Repair and Fracture Resistance in a Murine Myeloma Model

“…dose, route) or the pharmacokinetics and pharmacodynamics of each drug. For instance, there may be additional benefits to bone with SD‐208 due to off‐target effects, since SD‐208 also has a >25% inhibition of other kinases including AKT1‐3, protein kinase C (ε, η, and θ isoforms), epidermal growth factor receptor, protein kinase D, and mitogen‐activated protein kinase‐activated protein 2 …”

“…For instance, there may be additional benefits to bone with SD-208 due to off-target effects, since SD-208 also has a >25% inhibition of other kinases including AKT1-3, protein kinase C (ε, η, and θ isoforms), epidermal growth factor receptor, protein kinase D, and mitogen-activated protein kinaseactivated protein 2. (69,70) Bone anabolics show great promise in healing myeloma bone disease. Prevention of lesion development has been observed with anti-TGFβ therapies (1D11 (21,23) and now SD-208), an antisclerostin antibody, (58,71) an anti-DKK1 antibody, (72) and a soluble decoy receptor for activin A (73) in vivo.…”

Exquisitely Specific Bisubstrate Inhibitors of c-Src Kinase

“…We utilized a commercially available kinase panel of 213 kinases that measures competitive binding (KinaseSeeker TM , Luceome Biotechnologies). 18,19 In this panel, we found only two kinases (c-Src and its close homolog c-Yes) were significantly bound by bisubstrate inhibitor 3 at 115 nM, a concentration >400-fold higher than compound 3 ’s K d for c-Src (Figure 2). Notably, no other kinase that was found to be bound by ATP-competitive fragment 2 was potently bound by 3 , including other Src family kinases.…”