Table 6. Comparison of Profiles of Initial Hit 10 and Compounds 25 and 27.

specific measure	10	25	27
In Vitro
log kinact/Kia	4.43	4.60	4.92
WB CD69 IC50 [μM]b	0.333	0.079	0.036
TMD8 IC50 [nM]c	12.5	0.817	0.630
RLM/HLM [mL/min/kg]d	820/74	125/38	180/36
GSH t1/2 [h]e	9.4	4.3	3.2
fu (hPPB/rPPB/rbrainPB)f	0.13/0.29/0.16	0.10/0.08/0.04	0.05/0.12/0.13
MDR1-MDCK (A-B/ER)g	59/5.4	50/8.7	40/12.8
BCRP (A-B/ER)h	1.9	1.1	1.0
solubility pH 1.5, 7.5 [μg/mL]i	48	46	40
kinome selectivityj @1 μMk (S(10)-score)l	7/403 (0.017)	10/403 (0.025)	5/403 (0.012)
MW [g/mol]/Elog D/CNS MPOm	366.2/2.4/5.86	366.2/2.5/5.96	378.2/2.4/5.82
In Vivo	Rodentn
Kp,uu	0.37	0.11	0.44
Vdss [L/kg]	19.2	4.3	7.5
IV T1/2 [h]	10.3	3.59	0.74
CL [mL/min/kg]	133	87	121
F [%]	3	25	33
In Vivo	Nonrodento
Vdss [L/kg]	0.73	1.19	1.07
IV T1/2 [h]	0.21	0.29	0.37
CL [mL/min/kg]	34.2	53.5	37.0
F [%]v	<0.05	21	16

^ak_inact/K_i values were determined using the nonphosphorylated BTK protein in a continuous-read kinetic enzyme assay, with the mean value for replicates shown in parentheses.

^bMean IC₅₀ values for the inhibition of CD69 expression on CD19+ B cells in human whole blood upon stimulation with anti-IgD.

^cMean IC₅₀ for the inhibition of TMD8 B cell proliferation.

^dMetabolic stability in rat and human liver microsome (Intrinsic Clearance (Cl_int)).

^eIn vitro reactivity assessment using a GSH conjugation assay.

^f% Fraction unbound was measured at a concentration of 2 μM in human plasma via rapid equilibrium dialysis (RED).

^gEfflux ratio = P_app(B – A)/P_app(A – B) MDCK-MDR1.

^hEfflux ratio = P_app(B – A)/P_app(A – B) BCRP.

ⁱKinetic solubility was determined using an Analiza shake-flask solubility assay.

^jDiscoverX KINOMEScan Kinase Screen.

^kTotal number of kinases screened (403) excludes BTK.

^mMoKa was used to calculate clog P, clog D, and pK_a.

ⁿRodent PK studies were completed in male Sprague-Dawley (SD) rats, measured in triplicate.

^oNonrodent PK studies were completed in cynomolgus monkeys, measured in triplicate.