Abstract P5-04-23: Enhancing the activity of a novel estrogen receptor coregulator binding modulator (ERX-11) against ER-positive therapy resistant breast cancer

Abstract

Background:We had previously reported a novel small molecule, ERX-11, that directly interacts with ER and blocks the interaction between a subset of coregulators with both native and mutant forms of ER. ERX-11 effectively blocks ER oncogenic signaling and has potent anti-proliferative activity against therapy-sensitive and therapy-resistant human breast cancer cells. To enhance the clinical translation of ERX-11, we sought to pursue both lead optimization and evaluate combinations of ERX-11 with other approved agents in breast cancer. Methods: We designed, synthesized and tested 500 derivatives of ERX-11 in multiple models of ER+ breast cancer. We also tested combinations of ERX-11 with multiple agents, including other ER targeting agents, chemotherapies and CDK4/6 inhibitors. We tested the effect of combination therapy using breast cancer cells with acquired resistance (Tamoxifen, Letrozole, Ribociclib resistant) and engineered models that express ER mutations. In vitro activity was tested using Cell titer glo, MTT, and apoptosis assays. Mechanistic studies were conducted using Western blot, reporter gene assays and RNA-seq analysis. Xenograft, patient derived xenograft (PDX), patient derived explant (PDE) and xenograft derived explant (XDE) models were used for preclinical evaluation and toxicity. Result: Evaluation of 500 analogs of ERX-11 identified a number of leads with differential activity against ER+ and ER- breast cancer cells, identified several analogs including ERX-144, 208, 296, 315 with nanomolar potency against ER+ and therapy-resistant ER+ breast cancers. Validation of the mechanism of action of these analogs is ongoing. The combination of ERX-11 and palbociclib significantly blocked ER-mediated and ER-coregulators mediated oncogenic signaling and showed potent anti-proliferative activity against both endocrine therapy-sensitive and resistant breast cancer cells. In addition, ERX-11 inhibited ribociclib-resistant ER+ cell proliferation in a dose dependent manner. Mechanistic studies using IP-Mass spectrometry demonstrated that ERX-11 and palbociclib blocks the interaction between larger subset of coregulators with ER in therapy resistant breast cancer models. ERX-11 and palbociclib both exhibited potent anti-proliferative activity against therapy-sensitive and therapy-resistant ER+ve breast cancer cells, in xenograft models and in PDEs. Importantly, combination therapy of ERX-11 and palbociclib synergistically reduced the growth of tamoxifen and letrozole resistant xenograft tumors compared to either drug alone. Mass spec based DIA analyses and RNA-seq studies revealed that combinational treatment uniquely activated p53, unfolded response mediated apoptotic pathways, altered DNA damage response and suppressed E2F and Myc target genes. Biochemical studies confirmed combination therapy significantly altered E2F1, ER and DNA damage response pathways. Conclusion: We have successfully pursued two avenues to improving ERX-11 for clinical translation. We have developed ERX-11 analogs with higher potency against ER+ breast cancer. We have shown that combinational treatment with ERX-11 and palbociclib may overcome endocrine therapy resistance and CDK4/6 inhibitor (ribociclib) resistance. Citation Format: Viswanadhapalli S, Ma S, Lee T-K, Sareddy GR, Liu X, Ekoue D, Alluri A, Luo Y, Kassees K, Arteaga C, Alluri P, Weintraub SE, Tekmal RR, Ahn J-M, Raj GV, Vadlamudi RK. Enhancing the activity of a novel estrogen receptor coregulator binding modulator (ERX-11) against ER-positive therapy resistant breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P5-04-23.

Computer-Aided Ligand Discovery for Estrogen Receptor Alpha

“…ERX-11 in combination with a palbociclib (CDK4/6 inhibitor) was found to be more potent in tamoxifen and letrozole resistant cells than either treatment alone [287,288]. Lead optimization based on ERX-11 yielded four compounds with nanomolar activity against ERα, which are currently being validated in preclinical phase [287]. Essentially, the two iso-butane groups of ERX-11 mimic the LXXLL motif of the co-activator and fill up the volumes of leucine side chains of the LXXLL motif [284].…”

“…The compound also demonstrates significant activity in Tam-resistant and letrozole-resistant cell lines [284,286]. ERX-11 in combination with a palbociclib (CDK4/6 inhibitor) was found to be more potent in tamoxifen and letrozole resistant cells than either treatment alone [287,288]. Lead optimization based on ERX-11 yielded four compounds with nanomolar activity against ERα, which are currently being validated in preclinical phase [287].…”

“…(CDK4/6 inhibitor) was found to be more potent in tamoxifen and letrozole resistant cells than either treatment alone [287,288]. Lead optimization based on ERX-11 yielded four compounds with nanomolar activity against ERα, which are currently being validated in preclinical phase [287].…”

“…The use of SBDD with a larger chemical database to screen through could result in diverse active chemotypes and lead compounds that have not yet been explored. Although significant progress has been made in AF2-site drug discovery in recent years, until now only the empirically designed ERX-11 has reached later stages of drug development pipeline, presenting a promising candidate for successful inhibition of BCa cells (Table 6) [287,288]. However, we are yet to see a small molecule inhibitor that has reached the clinical trial phase.…”