Skip to main content
NCBI home page
NIHPA Author Manuscripts logoLink to NIHPA Author Manuscripts
. Author manuscript; available in PMC: 2026 May 16.
Published in final edited form as: Bioorg Chem. 2026 Feb 3;173:109601. doi: 10.1016/j.bioorg.2026.109601

An unprecedented potent inhibitor of MV4-11 cells: investigations into the mechanism of action beyond FLT3 inhibition

Xiuqi Wang a,b,1, Rosa Anna DeFilippis c, Huimin Geng d, Neil P Shah c, Hong-yu Li b,e,*
PMCID: PMC13178952  NIHMSID: NIHMS2160049  PMID: 41671740

Abstract

Activating mutations in FLT3 occur in 30% of acute myeloid leukemia (AML) cases. The AML patient-derived MV4–11 cell line contains a genetic alteration in FLT3 (“FLT3-ITD”), causing constitutive FLT3 activation. From screening, we identified compound 1 with unprecedently high anti-MV4–11 effects, with IC50 = 0.0021 ± 0.0003 nM. From the dose response curve, the effects of compound 1 are gradual and may have biphasic characteristics. Further studies identified compound 1 as a type-I FLT3 inhibitor with comparable potency to quizartinib and gilteritinib; however, compound 1 is much more potent against MV4–11 cells, indicating that it may have a second molecular mechanism of action independent of FLT3 inhibition. Interestingly, compound 1’s high potency is uniquely toward MV4–11 cells, and distinct from other cell lines either with or without FLT3 mutations. Preliminary efforts to unravel this mechanism were undertaken. The results of apoptosis assay and cell cycle analysis showed that the effects of compound 1 on MV4–11 may be biphasic, with an immediate cell cycle stabilizing effect at low picomolar concentrations, and a stronger effect to arrest cell cycle and induce apoptosis at low nanomolar concentrations. However, kinase selectivity profiling demonstrates that other than FLT3, compound 1 lacks strong binding affinities with other kinases. Therefore, the high inhibitory potency of compound 1 on MV4–11 cells appear unlikely to be due to synergism of co-inhibition of FLT3 and any other kinases. Altogether, compound 1 may serve as a promising lead compound for further optimization and research on a potentially new molecular antiproliferative mechanism.

Keywords: Acute myeloid leukemia, FLT3, mv4–11

1. Introduction

Acute myeloid leukemia (AML) is a hematologic cancer characterized by proliferative, clonal, abnormally differentiated, and occasionally poorly differentiated cells [1]. Activating mutations in FMS-like tyrosine kinase 3 (FLT3) are the most frequent genetic alteration in AML, found in approximately 30% of newly diagnosed cases. FLT3 activating mutations have two subtypes: internal tandem duplication at the juxtamembrane domain (FLT3-ITD) [2], and point mutations in the tyrosine kinase domain (FLT3-TKD) [3,4]. MV4–11 is an AML patient-derived cell line that harbors a FLT3-ITD mutation, and is often used to test the anti-FLT3 potency of kinase inhibitors. Three FLT3 inhibitors have been clinically approved – midostaurin, quizartinib [5,6], and gilteritinib [7]. However, these all have limitations. Therefore, improved FLT3 inhibitors are needed.

Imidazo[1,2-a]pyridine derivatives have previously been identified as potent antileukemic inhibitors [8,9]. In order to discover new active compounds to treat AML, we structurally optimized previously reported imidazo[1,2-a]pyridine derivatives [8,9] and performed structure-activity relationship (SAR) study in our previous research. We screened the imidazo[1,2-a]pyridine derivatives within our chemical library against the MV4–11 cell line. FLT3 inhibitors gilteritinib and quizartinib, and multi-kinase inhibitor vandetanib [10] were used as reference compounds (Fig. 1, Panel A). Through library screening, we identified N-(2,4-dimethoxyphenyl)-6-(7-(1-methyl-1H-pyrazol-4-yl) imidazo[1,2-a]pyridin-3-yl)pyridin-2-amine (compound 1) (Fig. 1, Panel B) with surprisingly low IC50 against MV4–11 cells, at 2.13 ± 0.29 pM.

Fig. 1.

Fig. 1.

Open in a new tab

The structure of compounds of interest. (A) Reference compounds gilteritinib, quizartinib, and vandetanib. (B) Compound 1. (C) FLT3 inhibitor MZH29 previously reported with high anti leukemic cellular effects.

Many small molecule compounds have shown substantial potency toward various cancer cell lines. In previous studies, inhibitors with high potency at mid- to high-picomolar levels against leukemic cell lines were described, and were usually associated with strong FLT3 inhibition [11]. For example, the FLT3 inhibitor MZH29 has high potency toward MV4–11 cells, with IC50 of 0.14 nM [12] (Fig. 1, Panel C). However, a compound with IC50 as low as ~2 pM as described here is unprecedented. More interestingly, while the IC50 of FLT3 inhibition of compound 1 is relatively low (16.26 ± 1.40 nM) it is not in the picomolar range, indicating that there may be another antiproliferative molecular mechanism distinct from FLT3 inhibition that is responsible for its high anti-MV4–11 potency. In order to understand the mechanism that made compound 1 so potent to MV4–11, we performed kinetic inhibition studies, western blot to assess target engagement, apoptosis assays, cell cycle analysis, and kinase selectivity profiling. Here, we report this unique phenomenon and preliminary mechanistic investigations.

2. Results and discussion

2.1. Compound 1’s high potency on MV4-11 cells is verified with dose-response curve

In order to further verify the surprisingly high potency of compound 1 against MV4-11 cells, we tested nine concentrations to establish a dose-response curve. In order to further investigate its antiproliferative mechanism, we compared its cellular dose-response curve to gilteritinib. In contrast to previously described FLT3 inhibitors, we noted that the effect of compound 1 against MV4–11 cells is gradual. For instance, gilteritinib has >90% inhibition on MV4–11 cell growth at 20 nM but <20% inhibition at 0.2 nM. In contrast, compound 1 has >90% inhibition at 20 nM while maintaining ~50% inhibition at 2 pM (Fig. 2). This biphasic dose-response curve indicates that compound 1’s effects may be more complex than simple FLT3 inhibition (Fig. S1).

Fig. 2.

Fig. 2.

Open in a new tab

Dose-response MV4–11 activity curve for compound 1 and gilteritinib.

2.2. Compound 1’s FLT3 inhibition is relatively lower

The growth inhibitory effects of FLT3 inhibitors against MV4–11 cells typically correlate with FLT3 kinase inhibitory effects. In order to assess if the antiproliferative effects of compound 1 on MV4–11 cells correlate with FLT3 inhibition, we used the ADP-Glo method to compare the extent of FLT3 kinase inhibition achieved with compound 1 with that of gilteritinib and quizartinib. The IC50s of these three compounds are comparable (Table 1). FLT3-ITD causes FLT3 autophosphorylation, leading to subsequent phosphorylation of downstream targets such as ERK, STAT5, and AKT. In order to further evaluate the FLT3 inhibitory effects on a cellular basis, we examined the status of signaling downstream of FLT3 in MV4–11 cells treated with compound 1 or quizartinib by western blot analysis. DMSO diluent was used as a treatment control in each experiment, and GAPDH as a protein loading control. Both quizartinib and compound 1 began achieving reduction of FLT3 autophosphorylation and downstream phosphorylation of ERK at ~1 nM (Fig. 3, Panel A and B). The experiment was performed in triplicate, and the FLT3 IC50 was calculated using levels of p-ERK (Table 1). The cell-based FLT3 inhibitory effects of compound 1 and quizartinib appear comparable (Fig. 3, Panel C).

Table 1.

Anti kinase activities (IC50s) of compounds of interest (nM).

Compound FLT3 (ADP-Glo)a FLT3 (Western blot)b
Gilteritinib 10.59 ± 1.42 N.T.c
Quizartinib 11.35 ± 2.97 0.66 ± 0.24
Vandetanib >1000 N.T.c
1 16.26 ± 1.40 0.80 ± 0.23
Open in a new tab
a

IC50 for ADP-Glo, compound concentration required to inhibit the luminescent signal by 50%, referring the ADP that is produced in the enzymatic reaction.

b

IC50 for western blot, compound concentration required to inhibit the p-ERK signal relative to untreated MV4–11 cells by 50%.

c

NT, not tested. Data are expressed as the mean ± SD from the dose–response curves of at least three independent experiments.

Fig. 3.

Fig. 3.

Open in a new tab

Western blot analysis of MV4–11 cells treated with quizartinib (A) and compound 1 (B). DMSO diluent was used as control in each experiment, and GAPDH as a loading control. (C) Dose-response quantification of p-ERK signal versus vehicle (DMSO) treatment.

Although the in vitro FLT3 kinase inhibitory effects are similar, inhibitors with different binding mechanisms may work differently in cells [13,14]. Therefore, it is possible that compound 1’s high potency toward MV4–11 cells is associated with a new FLT3 inhibitory mechanism. In order to verify if compound 1 confers a new binding mechanism with FLT3 kinase, we conducted additional inhibition kinetics studies with compound 1 (Fig. 4). We found that compound 1 has ATP-competitive characteristics, and can therefore be classified as a type-I FLT3 inhibitor, similar to gilteritinib. Both ADP-Glo method and western blot analysis showed comparable FLT3 inhibition of compound 1, gilteritinib and quizartinib, but compound 1 had much more anti-MV4–11 effects, indicating that compound 1 may have a molecular mechanism for antiproliferative effects that is separate from FLT3 inhibition.

Fig. 4.

Fig. 4.

Open in a new tab

Lineweaver–Burk plot of FLT3 inhibition kinetic studies co-incubated with various concentrations of compound 1.

2.3. Compound 1’s high potency is uniquely observed with MV4–11 cells

We tested the antiproliferative effects of the compounds of interest on additional cell lines (Table 2). Compound 1 has good antiproliferative effects on Molm14, another FLT3-ITD expressing cell line, though is not as potent as what was described above in MV4–11 cells. Compound 1’s inhibitory effects on Molm14 more closely correspond with FLT3 inhibition. Compound 1 has comparatively little inhibitory effects on cell lines lacking FLT3 activating mutations (K562 and Ba/F3 cells). Interestingly, compound 1 and its close analogue compound 2 (Fig. S2) work very similarly on all other cells tested except MV4–11 cells (Table S1). The results of cell based anti-proliferative assays indicate that the high potency of compound 1 is at least partly associated with its inhibitory effects to FLT3 kinase. However, among all the cells tested, Compound 1’s high potency at low picomolar level is uniquely observed with MV4–11 cells.

Table 2.

Antiproliferative activities (IC50s) of compounds of interest (nM).

Compound MV4–11a Molm14 K562 Ba/F3
Gilteritinib 1.07 ± 0.08 5.47 ± 0.25 13,100 ± 2532 968 ± 32
Quizartinib 0.177 ± 0.033 N.T.b N.T.b N.T.b
Vandetanib 1195 ± 203 1108 ± 84 1535 ± 150 5663 ± 254
1 0.0021 ± 0.0003 1.42 ± 0.23 1583 ± 313 8467 ± 499
Open in a new tab
a

IC50 on cells, compound concentration required to inhibit tumor cell proliferation by 50%. Data are expressed as the mean ± SD from the dose–response curves of at least three independent experiments.

b

NT, not tested.

2.4. Mechanism investigation for compound 1’s high potency on MV4–11 cells

In order to further unravel the cellular mechanism responsible for this observation, MV4–11 cells were subjected to apoptosis assays after 72 h treatment with compound 1 (Fig. 5). The result shows that in the low concentration treatments (1 pM to 1 nM), early apoptotic cells (lower right quadrant) very gradually accumulate as the concentration rises (1.7% of DMSO treatment and 2.1%, 2.3%, 3.4%, and 4.8% of 1 pM to 1 nM, respectively); in comparison, with the higher concentration treatment (10 nM), both early apoptotic cells (17.3% of 10 nM treatment versus 4.8% of 1 nM treatment) and late apoptotic cells (upper right quadrant, 10.3% of 10 nM treatment versus 1.8% of 1 nM treatment) have a notably higher percentage. Compound 1 treated cells do not reach a substantial apoptotic percentage until nanomolar concentrations, concentrations at which it is inhibiting FLT3 kinase activity, indicating that the potency of compound 1 against MV4–11 cells at low picomolar concentrations are uncoupled from proapoptotic effects.

Fig. 5.

Fig. 5.

Open in a new tab

Apoptosis analysis of compound 1 on MV4–11 cell line.

We also conducted cell cycle analysis on MV4–11 cells treated with various concentrations of compound 1 for 72 h (Fig. 6). We observed a noticeable gap of cell percentage at G1 phase between the treatments of DMSO and 1 pM compound 1 (47.2% versus 52.4%). There is no obvious gap of cell percentage at G1 phase between the treatments of 1 pM, 10 pM, and 100 pM (52.5%, 53.5%, and 54.5%, respectively); however, obvious gaps of G1 percentage were observed between the treatments of 100 pM and 1 nM (54.5% versus 58.1%), and between 1 nM and 10 nM (58.1% versus 68.7%). There is also a substantial decrease of cell percentage at G2 phase comparing the treatments of 1 nM and 10 nM (8.93% to 3.01%). In summary, the effects of compound 1 on MV4–11 cell cycle may have a biphasic characteristic, with an immediate cell cycle G1 phase stabilizing effect at 1 pM, and a stronger effect on cell cycle arrest at low nanomolar concentrations. Although there seems to be only about 10% difference in the percentage of cells in G1 phase treated with 1 pM compound 1 comparing to DMSO treatment, the difference in the final cell number may be much wider, due to the exponential effect during 3–4 cell cycles. Therefore, the results of cell cycle arrest analysis of compound 1 on MV4–11 cells correspond with its dose-response curve on MV4–11 inhibition. In comparison, in our control cell line MOLM14, no biphasic effects were observed. A significant difference in cell cycle was only observed at a concentration of 1 nM and 10 nM (Fig. S3).

Fig. 6.

Fig. 6.

Open in a new tab

Cell cycle analysis of compound 1 on MV4–11 cell line.

Therefore, it is possible that compound 1 has a molecular mechanism distinct from FLT3 inhibition at low picomolar concentrations. Simultaneous inhibition of FLT3 and other kinase targets, such as CDK9 [15,16], BCL-2 [17], PI3K [18], PDGFR [19], KIT [20], etc., has been shown to enhance antitumor effects [21]. In order to investigate if the high anti-MV4–11 potency of compound 1 is associated with coinhibition of other kinases, we performed kinase selectivity profiling. From the results of the site-directed competition binding assay of compound 1 on 96 kinases, 3 interactions (>50% wash-off, Kd < 1 nM) and 8 interactions >50% wash-off, Kd < 10 nM) were mapped for 1 nM and for 10 nM, respectively (Fig. 7). However, aside from FLT3, none of the kinases tested were mapped with >90% wash-off in either concentration. The results of kinase selectivity profiling shows that out of 96 kinases tested, no kinase other than FLT3 is strongly bound by compound 1, which suggests that its high potency is unlikely due to the effect of co-inhibition of FLT3 and another kinase.

Fig. 7.

Fig. 7.

Open in a new tab

Kinase selectivity profiling of compound 1.

MV4–11 was established from blasts cells of a 10-year-old male with biphenotypic B-myelomonocytic leukemia (AML FAB M5) that carries a FLT3-ITD mutation [22]. In order to generate some hypotheses on the molecular mechanism of compound 1’s anti-MV4–11 effects, we compared the gene fusion, gene mutation and CRISPR knockout data of MV4–11 with MOLM14. The fusion, mutation and CRISPR data were downloaded from the Cancer Dependency Map Portal (DepMap) Public 23Q2 release (https://depmap.org/portal/download/all/) for the two cell lines MV4–11 and MOLM14. In MOLM14 cells, only one fusion was detected, MLL-AF9, while in MV4–11 20 fusions were detected, including MLL-AF4 (Table S2). More mutations were detected in MOLM14 cells (541 mutations on 510 genes) than in MV411 cells (242 mutations on 194 genes). Only 21 genes were mutated in both cell lines (Fig. S4, Table S3). For genetic loss-of-function study, DepMap using genome-scale CRISPR-Cas9 genetic perturbation reagents to silence or knockout individual genes identified genes that affect cell survival. We downloaded gene effect estimates and z-scores (normalized per cell line) for all genes for the two cell lines and compared them side by side (Table S4), especially on the kinases from our compound experiments (Table S5). For gene effects, a score of less than − 0.5 represents depletion in most cell lines, while less than − 1 represents strong killing. A positive gene effect score indicates that when knocking out the gene, the cell line grows faster. From the 96 kinases screened, no kinase was observed to have strong killing of MV4–11 while have no killing of MOLM14 when knocked out. For other non-kinase genes, many were observed to have different Z scores with MV4–11 and MOLM14, representing differences in antiproliferative effects when knocked out. Therefore, these genes may also cause different antiproliferative effects in MV4–11 and MOLM14 when pharmacologically inhibited. Many of these genes were reported to be associated with cancer; multiple genes such as CSTF2T [23] and SMYD5 [24] influence cell cycle by affecting histone activity. Compound 1 may therefore be achieving its anti-proliferative activity in MV4–11 cells through inhibition of some of these proteins.

3. Conclusion

Through library screening, we identified N-(2,4-dimethoxyphenyl)- 6-(7-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyridin-3-yl)pyridin-2-amine (compound 1) with strikingly potent anti-MV4–11 activity. Although there have been previous reports of several potent antileukemic inhibitors [11,12], an inhibitor with an IC50 of 2 pM in a cell-based assay is unprecedented to the best of our knowledge. Further studies showed that it is a type-I FLT3 inhibitor. Surprisingly, although it achieves FLT3 inhibition with comparable potency as gilteritinib and quizartinib, it is much more effective at inhibiting the growth of MV4–11 cells, suggesting a possible novel antiproliferative mechanism. Our studies showed that the effects of compound 1 on MV4–11 may be biphasic, with an immediate cell cycle stabilizing effect in G1 phase at low picomolar concentrations, and a stronger effect on cell cycle arrest and induction of apoptosis at low nanomolar concentrations. With respect to kinases, aside from FLT3, compound 1 does not appear to have strong binding affinities with other kinases. Therefore, the high inhibitory potency of compound 1 on MV4–11 cells appears unlikely to be due to synergism of co-inhibition of FLT3 and any other kinases. The molecular mechanism of its high anti-MV4–11 effects remains unclear and the subject of ongoing investigation. Computational studies demonstrate several potential genes that differentially impact the viability of MV4–11 cells. Previous publications also indicate that besides kinases, FLT3 inhibition synergizes with multiple targets to exert antiproliferation effects, such as BCL-2, SHP2, eIF4a, and HDACs [25–31]. Compound 1 therefore represents a promising lead compound for further optimization and research into a potentially novel antiproliferative mechanism. With respect to clinical relevance, compound 1’s high potency appears to be restricted toward MV4–11 cells, although only a limited number of cell lines have been assessed to date. Further research may reveal the basis for the high sensitivity of MV4–11 cells to compound 1, and therefore benefit not only a subgroup of AML patients, but potentially those with other cancers as well.

Supplementary Material

Supplementary Data

Acknowledgements

For completion of this work, H. L. & N. S. were supported by the grants (NIHR01 CA249282). We also acknowledge Dr. Jungang Chen for his guidance in operating flow cytometry in the apoptosis assay. The cell cycle analysis was assisted by the UAMS Flow Cytometry Core Facility. The cell cycle analysis study was supported in part by the Center for Microbial Pathogenesis and Host Inflammatory Responses grant P20GM103625 through the NIH National Institute of General Medical Sciences Centers of Biomedical Research Excellence. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.bioorg.2026.109601.

Footnotes

CRediT authorship contribution statement

Xiuqi Wang: Writing – original draft, Data curation. Rosa Anna DeFilippis: Writing – review & editing, Data curation. Huimin Geng: Data curation. Neil P. Shah: Writing – review & editing. Hong-yu Li: Writing – review & editing, Conceptualization.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Data availability

Data will be made available on request.

References

  • [1].Ley TJ, Miller C, Ding L, Raphael BJ, Mungall AJ, Robertson A, Hoadley K, Triche TJ Jr, Laird PW, Baty JD, Fulton LL, Fulton R, Heath SE, Kalicki- Veizer J, Kandoth C, Klco JM, Koboldt DC, Kanchi KL, Kulkarni S, Lamprecht TL, Larson DE, Lin L, Lu C, McLellan MD, McMichael JF, Payton J, Schmidt H, Spencer DH, Tomasson MH, Wallis JW, Wartman LD, Watson MA, Welch J, Wendl MC, Ally A, Balasundaram M, Birol I, Butterfield Y, Chiu R, Chu A, Chuah E, Chun HJ, Corbett R, Dhalla N, Guin R, He A, Hirst C, Hirst M, Holt RA, Jones S, Karsan A, Lee D, Li HI, Marra MA, Mayo M, Moore RA, Mungall K, Parker J, Pleasance E, Plettner P, Schein J, Stoll D, Swanson L, Tam A, Thiessen N, Varhol R, Wye N, Zhao Y, Gabriel S, Getz G, Sougnez C, Zou L, Leiserson MD, Vandin F, Wu HT, Applebaum F, Baylin SB, Akbani R, Broom BM, Chen K, Motter TC, Nguyen K, Weinstein JN, Zhang N, Ferguson ML, Adams C, Black A, Bowen J, Gastier-Foster J, Grossman T, Lichtenberg T, Wise L, Davidsen T, Demchok JA, Shaw KR, Sheth M, Sofia HJ, Yang L, Downing JR, Eley G, Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia, N. Engl. J. Med 368 (2013) 2059–2074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [2].Smith CC, Wang Q, Chin CS, Salerno S, Damon LE, Levis MJ, Perl AE, Travers KJ, Wang S, Hunt JP, Zarrinkar PP, Schadt EE, Kasarskis A, Kuriyan J, Shah NP, Validation of ITD mutations in FLT3 as a therapeutic target in human acute myeloid leukaemia, Nature 485 (2012) 260–263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [3].Smith CC, Zhang C, Lin KC, Lasater EA, Zhang Y, Massi E, Damon LE, Pendleton M, Bashir A, Sebra R, Perl A, Kasarskis A, Shellooe R, Tsang G, Carias H, Powell B, Burton EA, Matusow B, Zhang J, Spevak W, Ibrahim PN, Le MH, Hsu HH, Habets G, West BL, Bollag G, Shah NP, Characterizing and overriding the structural mechanism of the Quizartinib-resistant FLT3 “gatekeeper” F691L mutation with PLX3397, Cancer Discov. 5 (2015) 668–679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [4].Smith CC, Lin K, Stecula A, Sali A, Shah NP, FLT3 D835 mutations confer differential resistance to type II FLT3 inhibitors, Leukemia 29 (2015) 2390–2392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [5].Zarrinkar PP, Gunawardane RN, Cramer MD, Gardner MF, Brigham D, Belli B, Karaman MW, Pratz KW, Pallares G, Chao Q, Sprankle KG, Patel HK, Levis M, Armstrong RC, James J, Bhagwat SS, AC220 is a uniquely potent and selective inhibitor of FLT3 for the treatment of acute myeloid leukemia (AML), Blood 114 (2009) 2984–2992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [6].Cortes J, Perl AE, Dōhner H, Kantarjian H, Martinelli G, Kovacsovics T, Rousselot P, Steffen B, Dombret H, Estey E, Strickland S, Altman JK, Baldus CD, Burnett A, Krämer A, Russell N, Shah NP, Smith CC, Wang ES, Ifrah N, Gammon G, Trone D, Lazzaretto D, Levis M, Quizartinib, an FLT3 inhibitor, as monotherapy in patients with relapsed or refractory acute myeloid leukaemia: an open-label, multicentre, single-arm, phase 2 trial, Lancet Oncol. 19 (2018) 889–903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [7].Mori M, Kaneko N, Ueno Y, Yamada M, Tanaka R, Saito R, Shimada I, Mori K, Kuromitsu S, Gilteritinib, a FLT3/AXL inhibitor, shows antileukemic activity in mouse models of FLT3 mutated acute myeloid leukemia, Investig. New Drugs 35 (2017) 556–565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [8].Frett B, McConnell N, Smith CC, Wang Y, Shah NP, Li HY, Computer aided drug discovery of highly ligand efficient, low molecular weight imidazopyridine analogs as FLT3 inhibitors, Eur. J. Med. Chem 94 (2015) 123–131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [9].Zhang L, Lakkaniga NR, Bharate JB, McConnell N, Wang X, Kharbanda A, Leung YK, Frett B, Shah NP, Li HY, Discovery of imidazo[1,2-a]pyridine-thiophene derivatives as FLT3 and FLT3 mutants inhibitors for acute myeloid leukemia through structure-based optimization of an NEK2 inhibitor, Eur. J. Med. Chem 225 (2021) 113776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [10].Koch L, Pharmacotherapy: vandetanib-a new therapeutic option in advanced medullary thyroid cancer, Nat. Rev. Endocrinol 8 (2012) 1. [DOI] [PubMed] [Google Scholar]
  • [11].Wang Z, Cai J, Cheng J, Yang W, Zhu Y, Li H, Lu T, Chen Y, Lu S, FLT3 inhibitors in acute myeloid leukemia: challenges and recent developments in overcoming resistance, J. Med. Chem 64 (2021) 2878–2900. [DOI] [PubMed] [Google Scholar]
  • [12].Xu B, Zhao Y, Wang X, Gong P, Ge W, MZH29 is a novel potent inhibitor that overcomes drug resistance FLT3 mutations in acute myeloid leukemia, Leukemia 31 (2017) 913–921. [DOI] [PubMed] [Google Scholar]
  • [13].Alotaibi AS, Yilmaz M, Kanagal-Shamanna R, Loghavi S, Kadia TM, DiNardo CD, Borthakur G, Konopleva M, Pierce SA, Wang SA, Tang G, Guerra V, Samra B, Pemmaraju N, Jabbour E, Short NJ, Issa GC, Ohanian M, Garcia-Manero G, Bhalla KN, Patel KP, Takahashi K, Andreeff M, Cortes JE, Kantarjian HM, Ravandi F, Daver N, Patterns of resistance differ in patients with acute myeloid leukemia treated with type I versus type II FLT3 inhibitors, Blood Cancer Discovery 2 (2021) 125–134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [14].Roskoski R Jr., Classification of small molecule protein kinase inhibitors based upon the structures of their drug-enzyme complexes, Pharmacol. Res 103 (2016) 26–48. [DOI] [PubMed] [Google Scholar]
  • [15].Reznickova E, Krajcovicova S, Perina M, Kovalova M, Soural M, Krystof V, Modulation of FLT3-ITD and CDK9 in acute myeloid leukaemia cells by novel proteolysis targeting chimera (PROTAC), Eur. J. Med. Chem 243 (2022) 114792. [DOI] [PubMed] [Google Scholar]
  • [16].Anshabo AT, Bantie L, Diab S, Lenjisa J, Kebede A, Long Y, Heinemann G, Karanjia J, Noll B, Basnet SKC, Li M, Milne R, Albrecht H, Wang S, An orally bioavailable and highly efficacious inhibitor of CDK9/FLT3 for the treatment of acute myeloid leukemia, Cancers (Basel) 14 (2022) 1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [17].Zhu R, Li L, Nguyen B, Seo J, Wu M, Seale T, Levis M, Duffield A, Hu Y, Small D, FLT3 tyrosine kinase inhibitors synergize with BCL-2 inhibition to eliminate FLT3/ITD acute leukemia cells through BIM activation, Signal Transduct. Target. Ther 6 (2021) 186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [18].Darici S, Zavatti M, Braglia L, Accordi B, Serafin V, Horne GA, Manzoli L, Palumbo C, Huang X, Jorgensen HG, Marmiroli S, Synergistic cytotoxicity of dual PI3K/mTOR and FLT3 inhibition in FLT3-ITD AML cells, Adv. Biol. Regul 82 (2021) 100830. [DOI] [PubMed] [Google Scholar]
  • [19].Smith CC, Lasater EA, Lin KC, Wang Q, McCreery MQ, Stewart WK, Damon LE, Perl AE, Jeschke GR, Sugita M, Carroll M, Kogan SC, Kuriyan J, Shah NP, Crenolanib is a selective type I pan-FLT3 inhibitor, Proc. Natl. Acad. Sci. USA 111 (2014) 5319–5324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [20].Warkentin AA, Lopez MS, Lasater EA, Lin K, He BL, Leung AY, Smith CC, Shah NP, Shokat KM, Overcoming myelosuppression due to synthetic lethal toxicity for FLT3-targeted acute myeloid leukemia therapy, Elife 3 (2014) e03445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [21].Peng FW, Xuan J, Wu TT, Xue JY, Ren ZW, Liu DK, Wang XQ, Chen XH, Zhang JW, Xu YG, Shi L, Design, synthesis and biological evaluation of N-phenylquinazolin-4-amine hybrids as dual inhibitors of VEGFR-2 and HDAC, Eur. J. Med. Chem 109 (2016) 1–12. [DOI] [PubMed] [Google Scholar]
  • [22].Asmaa MJS, Al-Jamal HA, Hussein AR, Yahaya BH, Hassan R, Hussain FA, Shamsuddin S, Johan MF, Transcriptomic profiles of MV4-11 and Kasumi 1 acute myeloid leukemia cell lines modulated by epigenetic modifiers Trichostatin a and 5-Azacytidine, Int. J. Hematol. Oncol. Stem Cell Res 14 (2020) 72–92. [PMC free article] [PubMed] [Google Scholar]
  • [23].Grozdanov PN, Li J, Yu P, Yan W, MacDonald CC, Cstf2t regulates expression of histones and histone-like proteins in male germ cells, Andrology 6 (2018) 605–615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [24].Zhang Y, Fang Y, Tang Y, Han S, Jia J, Wan X, Chen J, Yuan Y, Zhao B, Fang D, SMYD5 catalyzes histone H3 lysine 36 trimethylation at promoters, Nat. Commun 13 (2022) 3190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [25].Yao M, Yan W, Wang Y, Zhao Y, Xu X, Chen Y, Yu C, Li Y, Jiang H, Shen J, Cheng J, Xie C, IHCH9033, a novel class I HDAC inhibitor, synergizes with FLT3 inhibitor and rescues quizartinib resistance in FLT3-ITD AML via enhancing DNA damage response, Exp. Hematol. Oncol 14 (2025) 15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [26].Tan Y, Xin L, Wang Q, Xu R, Tong X, Chen G, Ma L, Yang F, Jiang H, Zhang N, Wu J, Li X, Guo X, Wang C, Zhou H, Zhou F, FLT3-selective PROTAC: enhanced safety and increased synergy with Venetoclax in FLT3-ITD mutated acute myeloid leukemia, Cancer Lett. 592 (2024) 216933. [DOI] [PubMed] [Google Scholar]
  • [27].Popescu B, Stahlhut C, Tarver TC, Wishner S, Lee BJ, Peretz CAC, Luck C, Phojanakong P, Camara Serrano JA, Hongo H, Rivera JM, Xirenayi S, Chukinas JA, Steri V, Tasian SK, Stieglitz E, Smith CC, Allosteric SHP2 inhibition increases apoptotic dependency on BCL2 and synergizes with venetoclax in FLT3- and KIT-mutant AML, Cell Rep. Med 4 (2023) 101290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [28].Janssen M, Schmidt C, Bruch PM, Blank MF, Rohde C, Waclawiczek A, Heid D, Renders S, Göllner S, Vierbaum L, Besenbeck B, Herbst SA, Knoll M, Kolb C, Przybylla A, Weidenauer K, Ludwig AK, Fabre M, Gu M, Schlenk RF, Stölzel F, Bornhäuser M, Röllig C, Platzbecker U, Baldus C, Serve H, Sauer T, Raffel S, Pabst C, Vassiliou G, Vick B, Jeremias I, Trumpp A, Krijgsveld J, Müller-Tidow C, Dietrich S, Venetoclax synergizes with gilteritinib in FLT3 wildtype high-risk acute myeloid leukemia by suppressing MCL-1, Blood 140 (2022) 2594–2610. [DOI] [PubMed] [Google Scholar]
  • [29].Nishida Y, Zhao R, Heese LE, Akiyama H, Patel S, Jaeger AM, Jacamo RO, Kojima K, Ma MCJ, Ruvolo VR, Chachad D, Devine W, Lindquist S, Davis RE, Porco JA Jr., Whitesell L, Andreeff M, Ishizawa J, Inhibition of translation initiation factor eIF4a inactivates heat shock factor 1 (HSF1) and exerts anti-leukemia activity in AML, Leukemia 35 (2021) 2469–2481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [30].Li D, Li T, Shang Z, Zhao L, Xu Q, Tan J, Qin Y, Zhang Y, Cao Y, Wang N, Huang L, Zhu X, Zhou K, Chen L, Li C, Xie T, Yang Y, Wang J, Zhou J, Combined inhibition of notch and FLT3 produces synergistic cytotoxic effects in FLT3/ITD(+) acute myeloid leukemia, Signal Transduct. Target. Ther 5 (2020) 21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [31].Brinton LT, Zhang P, Williams K, Canfield D, Orwick S, Sher S, Wasmuth R, Beaver L, Cempre C, Skinner J, Cannon M, Govande M, Harrington B, Lehman A, Byrd JC, Lapalombella R, Blachly JS, Synergistic effect of BCL2 and FLT3 co-inhibition in acute myeloid leukemia, J. Hematol. Oncol 13 (2020) 139. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Data
NIHMS2160049-supplement-Supplementary_Data.docx (557.5KB, docx)

Data Availability Statement

Data will be made available on request.

RESOURCES