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The novel protein homeostatic modulator BTX306 is active in myeloma and overcomes bortezomib and lenalidomide resistance

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Abstract

Small molecules targeting the cereblon-containing E3 ubiquitin ligase including thalidomide, lenalidomide, and pomalidomide modulate turnover of downstream client proteins and demonstrate pre-clinical and clinical anti-myeloma activity. Different drugs that engage with cereblon hold the potential of unique phenotypic effects, and we therefore studied the novel protein homeostatic modulator (PHM™) BTX306 with a unique thiophene-fused scaffold bearing a substituted phenylurea and glutarimide. This agent much more potently reduced human-derived myeloma cell line viability, with median inhibitory concentrations in the single nanomolar range versus micromolar values for lenalidomide or pomalidomide, and more potently activated caspases 3/8/9. While lenalidomide and pomalidomide induced greater degradation of Ikaros and Aiolos in myeloma cells, BTX306 more potently reduced levels of GSPT1, eRF1, CK1α, MCL-1, and c-MYC. Suppression of cereblon or overexpression of Aiolos or Ikaros induced relative resistance to BTX306, and this agent did not impact viability of murine hematopoietic cells in an in vivo model, demonstrating its specificity for human cereblon. Interestingly, BTX306 did show some reduced activity in lenalidomide-resistant cell line models but nonetheless retained its nanomolar potency in vitro, overcame bortezomib resistance, and was equipotent against otherwise isogenic cell line models with either wild-type or knockout TP53. Finally, BTX306 demonstrated strong activity against primary CD138-positive plasma cells, showed enhanced anti-proliferative activity in combination with bortezomib and dexamethasone, and was effective in an in vivo systemic model of multiple myeloma. Taken together, the data support further translational studies of BTX306 and its derivatives to the clinic for patients with relapsed and/or refractory myeloma.

Key messages

  • BTX306 has a unique thiophene-fused scaffold bearing phenylurea and glutarimide.

  • BTX306 is more potent against myeloma cells than lenalidomide or pomalidomide.

  • BTX306 overcomes myeloma cell resistance to lenalidomide or bortezomib in vitro.

  • BTX306 is active against primary myeloma cells, and shows efficacy in vivo.

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Abbreviations

CRBN:

Cereblon

GSPT1:

G to S phase transition 1

IKZF1:

Ikaros

IKZF3:

Aiolos

LEN:

Lenalidomide

MM:

Multiple myeloma

PHM:

Protein homeostatic modulator

POM:

Pomalidomide

RPPA:

Reverse phrase protein array

THAL:

Thalidomide

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Acknowledgments

The authors would like to thank the MD Anderson Flow Cytometry, and the RPPA and Imaging Core Facilities, both supported by the Cancer Center Support Grant (P30 CA16672), and thank Dr. Janet E. Mertz and Tawin Lempridee from the University of Wisconsin School of Medicine and Public Health for having provided Ikaros and Aiolos expression plasmids.

Funding

This work was supported by funding from BioTheryX, Inc. Additional support came from the National Cancer Institute (R01s CA184464 and CA194264), the Leukemia & Lymphoma Society Specialized Center of Research (SCOR-12206-17), and the Dr. Mirian and Sheldon G. Adelson Research Foundation.

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Authors and Affiliations

  1. Department of Lymphoma and Myeloma, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., Unit 429, Houston, TX, 77030, USA

    Jianxuan Zou, Richard J. Jones, Hua Wang, Isere Kuiatse, Fazal Shirazi, Elisabet E. Manasanch, Hans C. Lee & Robert Z. Orlowski

  2. BioTheryX Inc., San Diego, CA, USA

    Robert Sullivan, Leah Fung, Normand Richard, Paul Erdman, Eduardo Torres, Imelda Lam, Brooke McElwee, Aparajita H. Chourasia, Kyle W. H. Chan, Frank Mercurio & David I. Stirling

  3. School of Mathematics, Science & Engineering, Southwestern College, Chula Vista, CA, USA

    David Hecht

  4. Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Robert Z. Orlowski

Authors
  1. Jianxuan Zou
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  2. Richard J. Jones
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  3. Hua Wang
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Contributions

JZ planned and conducted most of the experiments, analyzed the results, and drafted the manuscript. JZ and FS performed animal experiments. RJJ, HW, and IK provided cell lines. HW provided technical support in lentiviral transduction of myeloma cells. RS, LF, NR, PE, ET, DH, IL, BM, and AHC planned and conducted compound development and revised the manuscript. EEM and HCL revised the manuscript. RJJ and KWHC participated in the design and analysis of experiments and reviewed the data and the manuscript. FM, DIS, and RZO conceived of the line of investigation, designed the experiments, analyzed the results, and edited the manuscript.

Corresponding author

Correspondence to Robert Z. Orlowski.

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Conflicts of interest

RS, LF, NR, PE, ET, IL, BM, AHC, KWHC, FM, and DIS are employees of BioTheryX, Inc., while the other authors have no financial conflicts of interest to declare with respect to BioTheryX, Inc.

EEM has received research support from Sanofi, Quest Diagnostics, Novartis, JW Pharma, and Merck, and consultant fees from Takeda, Celgene, Sanofi, Janssen, GSK, and Adaptive Biotechnologies. HCL has provided consultancy services to Amgen, Inc., Celgene, a wholly owned subsidiary of Bristol-Myers Squibb, GlaxoSmithKline, Janssen Pharmaceutical, Sanofi-Aventis, and Takeda Pharmaceutical, and has received research funding from Amgen, Inc., Celgene, a wholly owned subsidiary of Bristol-Myers Squibb, Daiichi Sankyo, GlaxoSmithKline, Janssen Pharmaceutical, and Takeda Pharmaceuticals. RZO declares laboratory research funding from BioTheryX, and clinical research funding from CARsgen Therapeutics, Celgene, Exelixis, Janssen Biotech, Sanofi-Aventis, Takeda Pharmaceuticals North America, Inc. Also, RZO has served on advisory boards for Amgen, Inc., Bristol-Myers Squibb, Celgene, EcoR1 Capital LLC, Forma Therapeutics, Genzyme, GSK Biologicals, Ionis Pharmaceuticals, Inc., Janssen Biotech, Juno Therapeutics, Kite Pharma, Legend Biotech USA, Molecular Partners, Regeneron Pharmaceuticals, Inc., Sanofi-Aventis, Servier, and Takeda Pharmaceuticals North America, Inc., and is a Founder of Asylia Therapeutics, Inc., with associated patents and an equity interest, though this technology does not bear on the current manuscript.

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Zou, J., Jones, R.J., Wang, H. et al. The novel protein homeostatic modulator BTX306 is active in myeloma and overcomes bortezomib and lenalidomide resistance. J Mol Med 98, 1161–1173 (2020). https://doi.org/10.1007/s00109-020-01943-6

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