American Society of Hematology

Figure 3

$Figure 3. The PIA assay is a valid surrogate of in vivo target inhibition for KW-2449. The immunoblot shows phospho-FLT3 (top) and total FLT3 (blot stripped and reprobed; bottom). A patient enrolled on the clinical trial of KW-2449 was administered a dose of 200 mg orally. Whole blood was obtained prior to the dose and at 2, 8, and 12 hours afterward, and separated into plasma and cellular fractions. Circulating blasts were isolated and analyzed for phospho-FLT3 as described in “Methods.” To perform the PIA assay, Molm14 cells were incubated for 1 hour in the plasma from the same time points, and likewise analyzed for phospho-FLT3 (as described in “Methods”). The first 4 lanes of the blot are from the circulating blasts, whereas the next 4 lanes show FLT3 from the Molm14 cells exposed to the plasma from which those blasts were isolated.$

The PIA assay is a valid surrogate of in vivo target inhibition for KW-2449. The immunoblot shows phospho-FLT3 (top) and total FLT3 (blot stripped and reprobed; bottom). A patient enrolled on the clinical trial of KW-2449 was administered a dose of 200 mg orally. Whole blood was obtained prior to the dose and at 2, 8, and 12 hours afterward, and separated into plasma and cellular fractions. Circulating blasts were isolated and analyzed for phospho-FLT3 as described in “Methods.” To perform the PIA assay, Molm14 cells were incubated for 1 hour in the plasma from the same time points, and likewise analyzed for phospho-FLT3 (as described in “Methods”). The first 4 lanes of the blot are from the circulating blasts, whereas the next 4 lanes show FLT3 from the Molm14 cells exposed to the plasma from which those blasts were isolated.

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