Alkynyl nicotinamides show antileukemic activity in drug-resistant acute myeloid leukemia
Baskar Ramdas, Neetu Dayal, Ruchi Pandey, Elizabeth Larocque, Rahul Kanumuri, Santhosh Kumar Pasupuleti, Sheng Liu, Chrysi Kanellopoulou, Elizabeth Fei Yin Chu, Rodrigo Mohallem, Saniya Virani, Gaurav Chopra, Uma K. Aryal, Rena Lapidus, Jun Wan, Ashkan Emadi, Laura S. Haneline, Frederick W. Holtsberg, M. Javad Aman, Herman O. Sintim, Reuben Kapur
Baskar Ramdas, Neetu Dayal, Ruchi Pandey, Elizabeth Larocque, Rahul Kanumuri, Santhosh Kumar Pasupuleti, Sheng Liu, Chrysi Kanellopoulou, Elizabeth Fei Yin Chu, Rodrigo Mohallem, Saniya Virani, Gaurav Chopra, Uma K. Aryal, Rena Lapidus, Jun Wan, Ashkan Emadi, Laura S. Haneline, Frederick W. Holtsberg, M. Javad Aman, Herman O. Sintim, Reuben Kapur
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Research Article Hematology

Alkynyl nicotinamides show antileukemic activity in drug-resistant acute myeloid leukemia

Abstract

Activating mutations of FLT3 contribute to deregulated hematopoietic stem and progenitor cell (HSC/Ps) growth and survival in patients with acute myeloid leukemia (AML), leading to poor overall survival. AML patients treated with investigational drugs targeting mutant FLT3, including Quizartinib and Crenolanib, develop resistance to these drugs. Development of resistance is largely due to acquisition of cooccurring mutations and activation of additional survival pathways, as well as emergence of additional FLT3 mutations. Despite the high prevalence of FLT3 mutations and their clinical significance in AML, there are few targeted therapeutic options available. We have identified 2 novel nicotinamide-based FLT3 inhibitors (HSN608 and HSN748) that target FLT3 mutations at subnanomolar concentrations and are potently effective against drug-resistant secondary mutations of FLT3. These compounds show antileukemic activity against FLT3ITD in drug-resistant AML, relapsed/refractory AML, and in AML bearing a combination of epigenetic mutations of TET2 along with FLT3ITD. We demonstrate that HSN748 outperformed the FDA-approved FLT3 inhibitor Gilteritinib in terms of inhibitory activity against FLT3ITD in vivo.

Authors

Baskar Ramdas, Neetu Dayal, Ruchi Pandey, Elizabeth Larocque, Rahul Kanumuri, Santhosh Kumar Pasupuleti, Sheng Liu, Chrysi Kanellopoulou, Elizabeth Fei Yin Chu, Rodrigo Mohallem, Saniya Virani, Gaurav Chopra, Uma K. Aryal, Rena Lapidus, Jun Wan, Ashkan Emadi, Laura S. Haneline, Frederick W. Holtsberg, M. Javad Aman, Herman O. Sintim, Reuben Kapur

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Figure 7

Superior growth-inhibitory effect of HSN748 compared with FDA-approved FLT3 inhibitor Gilteritinib on the development of AML in NSGS mice.

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Superior growth-inhibitory effect of HSN748 compared with FDA-approved F...
Figure shows PDX data from 2 different samples from patients with AML. Data from Panel A–G originated from the first sample from a patient with AML and Panels H-J from different patient sample. (A) Experimental design. Briefly, multimutational (FLT3 ITD(ins46), DNMT3AR882H, NPM1288FS12, and CHEK2) cells from patients with AML were transplanted to sublethally irradiated (200 rads) NSGS mice. A week after transplanting peripheral blood hCD45-positive cells engraftment was assessed, and, based on the engraftment, mice were divided into vehicle, Gilteritinib, and HSN748 groups randomly and followed up on treatment for 4 weeks. After treatment, mice were euthanized and assessed for hCD45 percentage in peripheral blood, spleen,and bone marrow. B shows the robust inhibitory effect of HSN748 on hCD45-positive cells in peripheral blood at indicated time points. Panel C shows the inhibitory effect of HSN748 on hCD45% in spleen. D shows spleen pictures and E shows the representative flow profiles of human and murine CD45-positive cells in bone marrow of vehicle, Gilteritinib, and HSN748-treated mice. X-axis showing hCD45-positive cells and y-axis showing mCD45-positive cells. (F) Quantification data on the frequency of hCD45 cells in the bone marrow in mice treated with various drugs. (G) Kaplan-Meier plot showing the effect of Gilteritinib and HSN748 treatment on the survival of same-patient sample-derived xenografts in a separate experiment (n = 8 in each group). (H) Dose dependent superior proliferation inhibitory effect of HSN748 and HSN608 compared with FDA-approved FLT3 inhibitor AC220 on different samples from patients with AML with a multimutation of DNMT3AR882C, FLT3ITD (E598–Y599ins12),N676K,Y842H, NRASG12D, and KMT2AMLL MLLPTD (exons 2–8) ending up relapsing profile. This patient had previously been on Gilteritinib and other kinase-inhibitor therapies. Panel I shows the same-patient sample-derived xenografts effect of treatment with HSN748 compared with Gilteritinib and vehicle on peripheral blood hCD45 frequency quantification. (J) Representative flow profiles of leukemic stem and progenitors (K) quantification data on the frequency of human leukemic stem and progenitors under drug treatment. Data represent median with interquartile range by ordinary 1-way ANOVA analysis. (n = 3–4 in each group ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05). Panel I shows a 5.5-week time point quantitative data on the effect of HSN748 on hCD45 frequency. A representative flow profile of hCD45 frequency on biweekly peripheral blood assessment for the impact of HSN748 treatment on engraftment and propagation of leukemic cells was presented in Supplemental Figure 11.