Abstract
Chronic myeloid leukaemia (CML) is driven by the activity of the BCR–ABL1 fusion oncoprotein. ABL1 kinase inhibitors have improved the clinical outcomes for patients with CML, with over 80% of patients treated with imatinib surviving for more than 10 years1. Second-generation ABL1 kinase inhibitors induce more potent molecular responses in both previously untreated and imatinib-resistant patients with CML2. Studies in patients with chronic-phase CML have shown that around 50% of patients who achieve and maintain undetectable BCR–ABL1 transcript levels for at least 2 years remain disease-free after the withdrawal of treatment3,4. Here we characterize ABL001 (asciminib), a potent and selective allosteric ABL1 inhibitor that is undergoing clinical development testing in patients with CML and Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukaemia. In contrast to catalytic-site ABL1 kinase inhibitors, ABL001 binds to the myristoyl pocket of ABL1 and induces the formation of an inactive kinase conformation. ABL001 and second-generation catalytic inhibitors have similar cellular potencies but distinct patterns of resistance mutations, with genetic barcoding studies revealing pre-existing clonal populations with no shared resistance between ABL001 and the catalytic inhibitor nilotinib. Consistent with this profile, acquired resistance was observed with single-agent therapy in mice; however, the combination of ABL001 and nilotinib led to complete disease control and eradicated CML xenograft tumours without recurrence after the cessation of treatment.
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Change history
29 March 2017
In the key to Figure 3b, ‘CME911’ was replaced with ‘ABL001’.
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Acknowledgements
The authors wish to thank the entire team who contributed to the discovery and development of ABL001.
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A.A.W., J.D., W.Z., S.Bu., A.Q.H. and M.P. directed or performed cell signalling, enzymology and/or genetic characterization work. J.S., A.L.M. and X.P. directed or performed medicinal chemistry work. W.J., S.W.C.-J. and P.F. directed or performed structural biology, NMR and/or structural modelling and cheminformatics analysis. A.A.W., A.L., S.Bu., F.L., V.I., G.B., S.D. and S.T. directed or analysed in vivo pharmacology, pharmacokinetic, formulation and/or safety studies. H.B. performed mathematical modelling. K.G.V., S.Br., D.M.R. and T.P.H. were responsible for clinical strategy and/or investigations. L.P., M.W., F.H., N.J.K. and W.R.S. contributed to overall project oversight and strategy. A.A.W. and W.R.S. wrote the manuscript.
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Extended data figures and tables
Extended Data Figure 1 Chemical structure of ABL001 and biophysical characterization of its binding.
a, Chemical structure of ABL001. b, NMR chemical shift assay to determine the location of ABL001 binding. c, NMR-based conformational assay using the resonance of Val525 to monitor the ‘bending’ of helix I in the presence and absence of ABL001. d, Isothermal calorimetry study to determine the binding affinity (Ka) of ABL001 to ABL1.
Extended Data Figure 2 Cellular activity of ABL001 relative to catalytic inhibitors of ABL1.
a, BCR–ABL1 48 h proliferation assay in Ba/F3 cells using a Britelite luciferase detection assay in the presence or absence of IL-3 across a dose range of ABL001 and nilotinib. Each assay was performed in quadruplicate; data are mean ± s.d. b, The sensitivity of KCL-22 cells to ABL001, nilotinib and dasatinib was determined in a 72-h growth assay. Each compound was tested in duplicate. c, KCL-22 cells were incubated with a range of compound concentrations for 1 h and immunoblots run to detect total STAT5 and pSTAT5 (Tyr694), total BCR–ABL1 and pBCR–ABL1 (Tyr245), total CRKL and pCRKL (Tyr207), and GAPDH as a loading control. d, Synergy studies were performed using ABL001 in combination with imatinib, nilotinib or dasatinib. KCL-22 cells were incubated with the compound combinations across a dose range for 72 h, and the level of cell growth relative to DMSO-treated cells was determined.
Extended Data Figure 3 Pharmacokinetics, pharmacodynamics and efficacy of ABL001.
a, Pharmacokinetic (PK) parameters of ABL001 in mouse, rat and dog after a single dose of ABL001. AUC, area under the curve; BA, bioavailability; CL, clearance; Cmax, maximum concentration observed; IV, intravenous; t1/2term, terminal half-life; PO, oral dosing; Tmax, time at maximum concentration; Vss, volume of distribution. b, Total plasma concentrations and levels of pSTAT5 (Tyr694) in fine needle aspirate samples taken from KCL-22 xenografts were monitored after a single oral administration of ABL001 at doses ranging from 3 to 30 mg kg−1. pSTAT5 (Tyr694) levels were determined using a pSTAT5 (Tyr694) meso scale discovery (MSD) assay with each sample run in duplicate; data are mean ± s.d. Samples are expressed as a percentage of the levels of pSTAT5 (Tyr694) before dosing (t = 0). c, ABL001 efficacy in KCL-22 xenograft tumours was assessed by monitoring tumour volume at doses ranging from 3 to 30 mg kg−1 on either a twice a day (BID) or once a day (QD) dosing schedules. Data are mean ± s.e.m. d, ABL001 efficacy in three patient-derived ALL systemic xenograft models (ALL-7015, AL-7119 and AL-7155) was assessed by FACS monitoring of the percentage of CD45+ cells per live cell in blood samples taken at varying time points after dosing with either 7.5 mg kg−1 BID (group 2) or 30 mg kg−1 BID (group 3) ABL001 for 3 weeks. A control group (group 1) was treated with PBS vehicle. Data are mean ± s.e.m. (n = 6 per group). e, The tolerability of increasing doses of ABL001 dosed on a BID schedule was determined by monitoring mouse body weight 2–3 times per week. Data are mean ± s.e.m. (n = 5 per group).
Extended Data Figure 4 Activity of ABL001 and nilotinib in KCL-22 cell clones expressing Thr315Ile and Ala337Val BCR–ABL1 variants.
a, The sensitivity of parental KCL-22 cells (WT) and KCL-22-resistant clones expressing BCR–ABL1 Ala337Val and Thr315Ile mutation variants to treatment with ABL001 (left) and nilotinib (right) was tested in 72 h growth assays. Samples were tested in duplicate; data are mean ± s.d. as a percentage of the vehicle-treated cells. b, KCL-22 Thr315Ile mutant cells were implanted as mouse xenografts and the efficacy of ABL001 across a dose range of 3–30 mg kg−1 BID was determined. Nilotinib was tested at 75 mg kg−1 BID as a control. Data are mean ± s.e.m. (n = 7 per group). T/C denotes ratio of tumour volume in control versus treated mice; ‘Reg’ denotes regression.
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Wylie, A., Schoepfer, J., Jahnke, W. et al. The allosteric inhibitor ABL001 enables dual targeting of BCR–ABL1. Nature 543, 733–737 (2017). https://doi.org/10.1038/nature21702
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DOI: https://doi.org/10.1038/nature21702
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