A selective BCL-XL PROTAC degrader achieves safe and potent antitumor activity
Sajid Khan, Xuan Zhang, Dongwen Lv, Qi Zhang, Yonghan He, Peiyi Zhang, Xingui Liu, Dinesh Thummuri, Yaxia Yuan, Janet S Wiegand, Jing Pei, Weizhou Zhang, Abhisheak Sharma, Christopher R McCurdy, Vinitha M Kuruvilla, Natalia Baran, Adolfo A Ferrando, Yong-Mi Kim, Anna Rogojina, Peter J Houghton, Guangcun Huang, Robert Hromas, Marina Konopleva, Guangrong Zheng, Daohong Zhou, Sajid Khan, Xuan Zhang, Dongwen Lv, Qi Zhang, Yonghan He, Peiyi Zhang, Xingui Liu, Dinesh Thummuri, Yaxia Yuan, Janet S Wiegand, Jing Pei, Weizhou Zhang, Abhisheak Sharma, Christopher R McCurdy, Vinitha M Kuruvilla, Natalia Baran, Adolfo A Ferrando, Yong-Mi Kim, Anna Rogojina, Peter J Houghton, Guangcun Huang, Robert Hromas, Marina Konopleva, Guangrong Zheng, Daohong Zhou
Abstract
B-cell lymphoma extra large (BCL-XL) is a well-validated cancer target. However, the on-target and dose-limiting thrombocytopenia limits the use of BCL-XL inhibitors, such as ABT263, as safe and effective anticancer agents. To reduce the toxicity of ABT263, we converted it into DT2216, a BCL-XL proteolysis-targeting chimera (PROTAC), that targets BCL-XL to the Von Hippel-Lindau (VHL) E3 ligase for degradation. We found that DT2216 was more potent against various BCL-XL-dependent leukemia and cancer cells but considerably less toxic to platelets than ABT263 in vitro because VHL is poorly expressed in platelets. In vivo, DT2216 effectively inhibits the growth of several xenograft tumors as a single agent or in combination with other chemotherapeutic agents, without causing appreciable thrombocytopenia. These findings demonstrate the potential to use PROTAC technology to reduce on-target drug toxicities and rescue the therapeutic potential of previously undruggable targets. Furthermore, DT2216 may be developed as a safe first-in-class anticancer agent targeting BCL-XL.
Conflict of interest statement
Competing interests: S.K., X.Z, Y.H., P.Z., G.Z., and D.Z. are inventors of two pending patent applications for use of BCL-XL PROTACs as senolytic and antitumor agents. R.H., G.Z., and D.Z. are co-founders of and have equity in Dialectic Therapeutics, which develops BCL-XL PROTACs to treat cancer.
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References
- Hanahan D & Weinberg RA Hallmarks of cancer: the next generation. Cell. 144, 646–674 (2011).
- Singh R, Letai A, & Sarosiek K Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nat Rev Mol Cell Biol. 20, 175–193 (2019).
- Igney FH & Krammer PH Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer. 2, 277–288 (2002).
- Ashkenazi A, Fairbrother WJ, Leverson JD, & Souers AJ From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors. Nat Rev Drug Discov. 16, 273–284 (2017).
- Adams JM & Cory S The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene. 26, 1324–1337 (2007).
- Reed JC Bcl-2-family proteins and hematologic malignancies: history and future prospects. Blood. 111, 3322–3330 (2008).
- Thomas S et al. Targeting the bcl-2 family for cancer therapy. Expert Opin Ther Targets. 17, 61–75 (2013).
- Opfermann JT Attacking cancer’s Achilles heel: antagonism of antiapoptotic BCL-2 family members. FEBS J. 283, 2661–2675 (2016).
- Garner TP, Lopez A, Reyna DE, Spitz AZ, & Gavathiotis E Progress in targeting the BCL-2 family of proteins. Curr Opin Chem Biol. 39, 133–142 (2017).
- Delbridge AR, Grabow S, Strasser A, & Vaux DL Thirty years of BCL-2: Translating cell death discoveries into novel cancer therapies. Nat Rev Cancer. 16, 99–109 (2016).
- Delbridge AR & Strasser A The BCL-2 protein family, BH3-mimetics and cancer therapy. Cell Death Differ. 22, 1071–1080 (2015).
- Oltersdorf T et al. An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature. 435, 677–681 (2005).
- Tse C et al. ABT-263: a potent and orally bioavailable Bcl-2 family inhibitor. Cancer Research. 68, 3421–3428 (2008).
- Souers AJ et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat Med. 19, 202–208 (2013).
- Tao ZF et al. Discovery of a potent and selective BCL-XL inhibitor with in vivo activity. ACS Med Chem Lett. 5, 1088–1093 (2014).
- Kotschy A et al. The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models. Nature. 538, 477–482 (2016).
- Deeks ED Venetoclax: First Global Approval. Drugs. 76, 979–987 (2016).
- Roberts AW et al. Targeting BCL2 with venetoclax in relapsed chronic lymphocytic leukemia. N Engl J Med. 374: 311–322 (2016).
- Mason KD et al. Programmed anuclear cell death delimits platelet life span. Cell. 128, 1173–1186 (2007).
- Schoenwaelder SM et al. Bcl-xL–inhibitory BH3 mimetics can induce a transient thrombocytopathy that undermines the hemostatic function of platelets. Blood. 118: 1663–1674 (2011).
- Kaefer A et al. Mechanism-based pharmacokinetic/pharmacodynamic meta-analysis of navitoclax (ABT-263) induced thrombocytopenia. Cancer Chemother Pharmacol. 74, 593–602 (2014).
- Itchaki G & Brown JR The potential of venetoclax (ABT-199) in chronic lymphocytic leukemia. The Adv Hematol. 7, 270–287 (2016).
- Perini GF, Ribeiro GN, Neto JVP, Campos LT, & Hamerschlak N BCL-2 as therapeutic target for hematological malignancies. J Hematol Oncol. 11, 65 (2018).
- Leverson JD et al. Exploiting selective BCL-2 family inhibitors to dissect cell survival dependencies and define improved strategies for cancer therapy. Sci Transl Med. 7, 279ra40 (2015).
- Amundson SA et al. An informatics approach identifying markers of chemosensitivity in human cancer cell lines. Cancer Res. 60, 6101–6110 (2000).
- Vogler M Targeting BCL2-proteins for the treatment of solid tumours. Adv in Medicine. 2014, 943648 (2014).
- Lai AC, & Crews CM Induced protein degradation: an emerging drug discovery paradigm. Nat Rev Drug Discov. 16, 101–114 (2017).
- Runcie AC, Chan KH, Zengerle M, & Ciulli A Chemical genetics approaches for selective intervention in epigenetics. Curr Opin Chem Biol. 33, 186–194 (2016).
- Deshaies RJ Protein degradation: Prime time for PROTACs. Nat Chem Biol. 11, 634–635 (2015).
- Churcher I Protac-induced protein degradation in drug discovery: breaking the rules or just making new ones? J Med Chem. 61, 444–452 (2018).
- Ohoka N, Shibata N, Hattori T, & Naito M Protein knockdown technology: application of ubiquitin ligase to cancer therapy. Curr Cancer Drug Targets. 16, 136–146 (2016).
- Lu J, et al. Hijacking the E3 ubiquitin ligase cereblon to efficiently target BRD4. Chem Biol. 22, 755–763 (2015).
- Bondeson DP et al. Catalytic in vivo protein knockdown by small-molecule PROTACs. Nat Chem Biol 11, 611–617 (2015).
- Lai AC et al. Modular PROTAC design for the degradation of oncogenic BCR-ABL. Angew Chem Int Ed. 55, 807–810 (2016).
- Raina K et al. PROTAC-induced BET protein degradation as a therapy for castration-resistant prostate cancer. Proc Natl Acad Sci USA. 113, 7124–7129 (2016).
- Saenz DT et al. Novel BET protein proteolysis-targeting chimera exerts superior lethal activity than bromodomain inhibitor (BETi) against post-myeloproliferative neoplasm secondary (s) AML cells. Leukemia. 1–11 (2017).
- Winter GE et al. DRUG DEVELOPMENT. Phthalimide conjugation as a strategy for in vivo target protein degradation. Science. 348, 1376–1381 (2015).
- Huang HT et al. A Chemoproteomic Approach to Query the Degradable Kinome Using a Multi-kinase Degrader. Cell Chem Biol. 25, 88–99.e6 (2018).
- Bray PF et al. The complex transcriptional landscape of the anucleate human platelet. BMC Genomics. 14, 1 (2013).
- Kissopoulou A, Jonasson J, Lindahl TL, & Osman A Next generation sequencing analysis of human platelet PolyA+ mRNAs and rRNA-depleted total RNA. PLoS One. 8, e81809 (2013).
- Cerami E et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2, 401–404 (2012).
- Gao J et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal. 6, pl1.
- Vogler M et al. BCL2/BCL-X(L) inhibition induces apoptosis, disrupts cellular calcium homeostasis, and prevents platelet activation. Blood. 117, 7145–7154 (2011).
- Gadd MS et al. Structural basis of PROTAC cooperative recognition for selective protein degradation. Nat Chem Biol. 13, 514–521 (2017).
- Nowak RP et al. Plasticity in binding confers selectivity in ligand-induced protein degradation. Nat Chem Biol. 14, 706–714 (2018).
- Riching KM et al. Quantitative Live-Cell Kinetic Degradation and Mechanistic Profiling of PROTAC Mode of Action. ACS Chem Biol. 13, 2758–2770 (2018).
- Farmer T, O’Neill KL, Naslavsky N, Luo X, & Caplan S Retromer facilitates the localization of Bcl-xL to the mitochondrial outer membrane. Molecular biology of the cell, 30(10), 1138–1146 (2019).
- Smith BE et al. Differential PROTAC substrate specificity dictated by orientation of recruited E3 ligase. Nat Commun. 10, 131 (2019).
- Morowski M et al. Only severe thrombocytopenia results in bleeding and defective thrombus formation in mice. Blood. 121, 4938–4947 (2013).
- Rinder HM et al. Correlation of thrombosis with increased platelet turnover in thrombocytosis. Blood. 91, 1288–1294 (1998).
- Koch R et al. Biomarker-driven strategy for MCL1 inhibition in T-cell lymphomas. Blood. 133, 566–575 (2018).
- Berger S et al. Computationally designed high specificity inhibitors delineate the roles of BCL2 family proteins in cancer. Elife. 5, pii: e20352 (2016).
- Hikita H et al. Mcl-1 and Bcl-xL cooperatively maintain integrity of hepatocytes in developing and adult murine liver. Hepatology. 50, 1217–1226.
- Chen J et al. The Bcl-2/Bcl-X(L)/Bcl-w inhibitor, navitoclax, enhances the activity of chemotherapeutic agents in vitro and in vivo. Mol Cancer Ther. 10, 2340–2349 (2011).
- Ackler S et al. The Bcl-2 inhibitor ABT-263 enhances the response of multiple chemotherapeutic regimens in hematologic tumors in vivo. Cancer Chemother Pharmacol. 66: 869–880 (2010).
- Pompili L, Porru M, Caruso C, Biroccio A, & Leonetti C Patient-derived xenografts: a relevant preclinical model for drug development. J Exp Clin Cancer Res. 35, 189 (2016).
- Zengerle M, Chan KH, & Ciulli A Selective Small Molecule Induced Degradation of the BET Bromodomain Protein BRD4. ACS Chem Biol. 10, 1770–1777 (2015).
- Bondeson DP et al. Lessons in PROTAC Design from Selective Degradation with a Promiscuous Warhead. Cell Chem Biol. 25, 78–87.e5 (2018).
- Lv D-W, Zhang K, & Li R Interferon regulatory factor 8 regulates aspase-1 expression to facilitate Epstein-Barr virus reactivation in response to B cell receptor stimulation and chemical induction. PLoS Pathog. 14, e1006868 ((2018).
- Wiśniewski JR et al. Universal sample preparation method for proteome analysis. Nat Methods. 6, 359–362 (2009).
- Nesvizhskii AI, Keller A, Kolker E & Aebersold R A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem. 75, 4646–4658 (2003).
- Tyanova S et al. The Perseus computational platform for comprehensive analysis of (prote) omics data. Nat Methods. 13, 731–740 (2016).
- Vizcaíno JA et al. The PRoteomics IDEntifications (PRIDE) database and associated tools: status in 2013. Nucleic Acids Res. 41, D1063–D1069 (2013).
- Food US and Administration Drug. “Bioanalytical Method Validation, Guidance for Industry.” US Department of Health and Human Services Food and Drug Administration, Rockville, MD: (2018).
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