Superior anti-tumor activity of the MDM2 antagonist idasanutlin and the Bcl-2 inhibitor venetoclax in p53 wild-type acute myeloid leukemia models

Abstract In acute myeloid leukemia (AML), TP53 mutations and dysregulation of wild-type p53 is common and supports an MDM2 antagonist as a therapy. RO6839921 is an inactive pegylated prodrug of the oral MDM2 antagonist idasanutlin (active principle [AP]) that allows for IV administration. This phase 1 monotherapy study evaluated the safety, pharmacokinetics, and pharmacodynamics of RO6839921 in patients with AML. Primary objectives identified dose-limiting toxicities (DLTs) and maximum tolerated dose (MTD). Secondary objectives assessed pharmacokinetic, pharmacodynamic, and antileukemic activity. A total of 26 patients received 120–300 mg AP of idasanutlin. The MTD was 200 mg, with DLTs at 250 (2/8 patients) and 300 mg (2/5). Treatment–related adverse events in >20% of patients were diarrhea, nausea, vomiting, decreased appetite, and fatigue. Six deaths (23.1%) occurred, all unrelated to treatment. Pharmacokinetics showed rapid and near-complete conversion of the prodrug to AP and dose-proportional exposure across doses. Variability ranged from 30%–47% (22%–54% for idasanutlin). TP53 was 21 (87.5%) wild-type and 3 mutant (12.5%). The composite response rate (complete remission [CR], CR with incomplete hematologic recovery/morphological leukemia-free state [CRi/MLFS], or CR without platelet recovery [CRp]) was 7.7%. Antileukemic activity (CR, CRi/MLFS, partial response, hematologic improvement/stable disease) was observed in 11 patients (disease control rate, 42%): 10/11 were TP53 wild-type; 1 had no sample. p53 activation was demonstrated by MIC-1 induction and was associated with AP exposure. There was not sufficient differentiation or improvement in the biologic or safety profile compared with oral idasanutlin to support continued development of RO6839921. NCT02098967.

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Phase I/II Study of Combination Therapy With Sorafenib, Idarubicin, and Cytarabine in Younger Patients With Acute Myeloid Leukemia

Journal of Clinical Oncology ◽

10.1200/jco.2009.25.4888 ◽

2010 ◽

Vol 28 (11) ◽

pp. 1856-1862 ◽

Cited By ~ 258

Author(s):

Farhad Ravandi ◽

Jorge E. Cortes ◽

Daniel Jones ◽

Stefan Faderl ◽

Guillermo Garcia-Manero ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Complete Remission ◽

Phase I ◽

Myeloid Leukemia ◽

Wild Type ◽

Platelet Recovery ◽

Probability Of Survival ◽

Target Effect ◽

Acute Myeloid

Purpose To determine the efficacy and toxicity of the combination of sorafenib, cytarabine, and idarubicin in patients with acute myeloid leukemia (AML) younger than age 65 years. Patients and Methods In the phase I part of the study, 10 patients with relapsed AML were treated with escalating doses of sorafenib with chemotherapy to establish the feasibility of the combination. We then treated 51 patients (median age, 53 years; range, 18 to 65 years) who had previously untreated AML with cytarabine at 1.5 g/m2 by continuous intravenous (IV) infusion daily for 4 days (3 days if > 60 years of age), idarubicin at 12 mg/m2 IV daily for 3 days, and sorafenib at 400 mg orally twice daily for 7 days. Results Overall, 38 (75%) patients have achieved a complete remission (CR), including 14 (93%) of 15 patients with mutated FMS-like tyrosine kinase-3 (FLT3; the 15th patient had complete remission with incomplete platelet recovery [CRp]) and 24 (66%) of 36 patients with FLT3 wild-type (WT) disease (three additional FLT3-WT patients had CRp). FLT3-mutated patients were more likely to achieve a CR than FLT3-WT patients (P = .033). With a median follow-up of 54 weeks (range, 8 to 87 weeks), the probability of survival at 1 year is 74%. Among the FLT3-mutated patients, 10 have relapsed and five remain in CR with a median follow-up of 62 weeks (range, 10 to 76 weeks). Plasma inhibitory assay demonstrated an on-target effect on FLT3 kinase activity. Conclusion Sorafenib can be safely combined with chemotherapy, produces a high CR rate in FLT3-mutated patients, and inhibits FLT3 signaling.

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High incidence of alternatively spliced forms of deoxycytidine kinase in patients with resistant acute myeloid leukemia

Blood ◽

10.1182/blood.v96.4.1517.h8001517_1517_1524 ◽

2000 ◽

Vol 96 (4) ◽

pp. 1517-1524 ◽

Cited By ~ 1

Author(s):

Marjan J. T. Veuger ◽

M. Willy Honders ◽

Jim E. Landegent ◽

Roel Willemze ◽

Renée M. Y. Barge

Keyword(s):

T Cells ◽

Acute Myeloid Leukemia ◽

Messenger Rna ◽

Myeloid Leukemia ◽

Deoxycytidine Kinase ◽

Wild Type ◽

Healthy Donors ◽

Alternatively Spliced ◽

Acute Myeloid

Deficiency of functional deoxycytidine kinase (dCK) is a common characteristic for in vitro resistance to cytarabine (AraC). To investigate whether dCK is also a target for induction of AraC resistance in patients with acute myeloid leukemia (AML), we determined dCK messenger RNA (mRNA) expression in (purified) leukemic blasts and phytohemagglutinin-stimulated T cells (PHA T cells) from patients with chemotherapy-sensitive and chemotherapy-resistant AML. In control samples from healthy donors (PHA T cells and bone marrow), only wild-type dCK complementary DNA (cDNA) was amplified. Also, in (purified) leukemic blasts from patients with sensitive AML, only wild-type dCK cDNAs were observed. These cDNAs coded for active dCK proteins in vitro. However, in 7 of 12 (purified) leukemic blast samples from patients with resistant AML, additional polymerase chain reaction fragments with a deletion of exon 5, exons 3 to 4, exons 3 to 6, or exons 2 to 6 were detected in coexpression with wild-type dCK. Deletion of exons 3 to 6 was also identified in 6 of 12 PHA T cells generated from the patients with resistant AML. The deleted dCK mRNAs were formed by alternative splicing and did code for inactive dCK proteins in vitro. These findings suggest that the presence of inactive, alternatively spliced dCK mRNA transcripts in resistant AML blasts may contribute to the process of AraC resistance in patients with AML.

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Detection of the JAK2V617F Mutation in Myeloproliferative Disorders by Melting Curve Analysis Using the LightCycler System

Archives of Pathology & Laboratory Medicine ◽

10.5858/2006-130-997-dotjmi ◽

2006 ◽

Vol 130 (7) ◽

pp. 997-1003

Author(s):

Randall J. Olsen ◽

Zhouwen Tang ◽

Daniel H. Farkas ◽

David W. Bernard ◽

Youli Zu ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

De Novo ◽

Clinical Laboratory ◽

Melting Curve ◽

Myeloproliferative Disorder ◽

Curve Analysis ◽

Melting Curve Analysis ◽

Wild Type ◽

Acute Myeloid

Abstract Context.—A specific mutation, JAK2V617F, was recently recognized as having diagnostic value for myeloproliferative disorders. No practical assay is currently available for routine use in a clinical laboratory. Objective.—We report the development of a real-time polymerase chain reaction melting curve analysis assay that is appropriate for molecular diagnostics testing. Design.—Specific primers and fluorescence resonance energy transfer probes were designed, and patients with a previously diagnosed myeloproliferative disorder, de novo acute myeloid leukemia, or reactive condition were selected. The DNA was extracted from fresh and archived peripheral blood and bone marrow specimens, and real-time polymerase chain reaction melting curve analysis was performed on the LightCycler platform (Roche Applied Science, Indianapolis, Ind). Results.—The JAK2 region was successfully amplified, and wild-type amplicons were reproducibly discriminated from JAK2V617F amplicons. Titration studies using homozygous wild-type and mutant cell lines showed the relative areas under a melting curve were proportional to allele proportion, and the assay reliably detected one mutant in 20 total cells. JAK2V617F was identified in patients previously diagnosed with a myeloproliferative disorder or acute myeloid leukemia transformed from myeloproliferative disorder, whereas a wild-type genotype was identified in patients with reactive conditions or de novo acute myeloid leukemia. Conclusions.—These findings demonstrate the suitability of this assay for identifying JAK2V617F in a clinical laboratory setting. Furthermore, the semiquantitative detection of JAK2V617F in archived specimens provides a new tool for studying the prognostic significance of this mutation.

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Preferential transcription of the mutated allele in NPM1 mutated acute myeloid leukaemia

Scientific Reports ◽

10.1038/s41598-020-73782-x ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

G. D. Bailey ◽

L. Doolan ◽

A. Baskar ◽

L. C. Smith ◽

C. H. Seedhouse

Keyword(s):

Acute Myeloid Leukemia ◽

Acute Myeloid Leukaemia ◽

Myeloid Leukaemia ◽

Myeloid Leukemia ◽

Splice Variants ◽

Wild Type ◽

Significant Bias ◽

Acute Myeloid

Abstract Nucleophosmin is commonly both over-expressed and mutated in acute myeloid leukemia (AML). NPM1 mutations are always heterozygous. In addition, NPM1 has a number of different splice variants with the major variant encoded by exons 1–9 and 11–12 (NPM1.1). Further variants include NPM1.2 which lacks exons 8 and 10 and NPM1.3 which comprises exons 1–10 (and so lacks the region of sequence mutated in AML). In this study we quantified the expression of these three variants in 108 AML patient samples with and without NPM1 mutations and also assessed the level of expression from the wild-type and mutant alleles in variants NPM1.1 and NPM1.2. The results show that NPM1.1 is the most commonly expressed variant, however transcripts from wild-type and mutated alleles do not occur at equal levels, with a significant bias toward the mutated allele. Considering the involvement of mutant nucleophosmin in the progression and maintenance of AML, a bias towards mutated transcripts could have a significant impact on disease maintenance.

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PF225 AZACITIDINE (AZA) ENHANCES ANTILEUKEMIC ACTIVITY OF THE MDM2 INHIBITOR MILADEMETAN IN TP53 WILD-TYPE ACUTE MYELOID LEUKEMIA (AML)

HemaSphere ◽

10.1097/01.hs9.0000559116.40081.dc ◽

2019 ◽

Vol 3 (S1) ◽

pp. 65 ◽

Cited By ~ 2

Author(s):

S. Noguchi ◽

T. Seki ◽

N. Adachi ◽

H. Sumi ◽

R. Nakano ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Wild Type ◽

Antileukemic Activity ◽

Mdm2 Inhibitor ◽

Acute Myeloid

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High incidence of alternatively spliced forms of deoxycytidine kinase in patients with resistant acute myeloid leukemia

Blood ◽

10.1182/blood.v96.4.1517 ◽

2000 ◽

Vol 96 (4) ◽

pp. 1517-1524 ◽

Cited By ~ 57

Author(s):

Marjan J. T. Veuger ◽

M. Willy Honders ◽

Jim E. Landegent ◽

Roel Willemze ◽

Renée M. Y. Barge

Keyword(s):

T Cells ◽

Acute Myeloid Leukemia ◽

Messenger Rna ◽

Myeloid Leukemia ◽

Deoxycytidine Kinase ◽

Wild Type ◽

Healthy Donors ◽

Alternatively Spliced ◽

Acute Myeloid

Abstract Deficiency of functional deoxycytidine kinase (dCK) is a common characteristic for in vitro resistance to cytarabine (AraC). To investigate whether dCK is also a target for induction of AraC resistance in patients with acute myeloid leukemia (AML), we determined dCK messenger RNA (mRNA) expression in (purified) leukemic blasts and phytohemagglutinin-stimulated T cells (PHA T cells) from patients with chemotherapy-sensitive and chemotherapy-resistant AML. In control samples from healthy donors (PHA T cells and bone marrow), only wild-type dCK complementary DNA (cDNA) was amplified. Also, in (purified) leukemic blasts from patients with sensitive AML, only wild-type dCK cDNAs were observed. These cDNAs coded for active dCK proteins in vitro. However, in 7 of 12 (purified) leukemic blast samples from patients with resistant AML, additional polymerase chain reaction fragments with a deletion of exon 5, exons 3 to 4, exons 3 to 6, or exons 2 to 6 were detected in coexpression with wild-type dCK. Deletion of exons 3 to 6 was also identified in 6 of 12 PHA T cells generated from the patients with resistant AML. The deleted dCK mRNAs were formed by alternative splicing and did code for inactive dCK proteins in vitro. These findings suggest that the presence of inactive, alternatively spliced dCK mRNA transcripts in resistant AML blasts may contribute to the process of AraC resistance in patients with AML.

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JAK2 V617F-Mutated Myeloproliferative Neoplasia Developing Five Years after Wild-Type JAK2 Acute Myeloid Leukemia: A Case Report

Acta Haematologica ◽

10.1159/000342100 ◽

2013 ◽

Vol 129 (1) ◽

pp. 23-25 ◽

Cited By ~ 1

Author(s):

Sabine Girsberger ◽

Axel Karow ◽

Pontus Lundberg ◽

Stephan Dirnhofer ◽

Thomas Lehmann ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Case Report ◽

Myeloid Leukemia ◽

Jak2 V617f ◽

Wild Type ◽

Acute Myeloid

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Recent advances in FLT3 inhibitors for acute myeloid leukemia

Future Medicinal Chemistry ◽

10.4155/fmc-2019-0365 ◽

2020 ◽

Vol 12 (10) ◽

pp. 961-981 ◽

Cited By ~ 1

Author(s):

Lexian Tong ◽

Xuemei Li ◽

Yongzhou Hu ◽

Tao Liu

Keyword(s):

Acute Myeloid Leukemia ◽

Tyrosine Kinase ◽

Myeloid Leukemia ◽

Clinical Stage ◽

Wild Type ◽

New Techniques ◽

Flt3 Mutations ◽

Flt3 Inhibitors ◽

Target Activity ◽

Acute Myeloid

Fms-like tyrosine kinase-3 (FLT3) mutations occur in approximately 30% of acute myeloid leukemia (AML) cases, suggesting FLT3 as an attractive target for AML treatment. Early FLT3 inhibitors enhance antileukemia efficacy by inhibiting multiple targets, and thus had stronger off-target activity, increasing their toxicity. Recently, a number of potent and selective FLT3 inhibitors have been developed, many of which are effective against multiple mutations. This review outlines the evolution of AML-targeting FLT3 inhibitors by focusing on their chemotypes, selectivity and activity over FLT3 wild-type and FLT3 mutations as well as new techniques related to FLT3. Compounds that currently enter the late clinical stage or have entered the market are also briefly reported.

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