Abstract 3086: Discovery of a first-in-class TEAD inhibitor which directly inhibits YAP/TAZ-TEAD protein-protein interaction and shows a potent anti-tumor effect in malignant pleural mesothelioma

Abstract

Objective: Hippo signaling pathway is known to regulate organ development. In Hippo signaling pathway, YAP or TAZ works as a transcriptional co-activator and forms a transcriptional complex with TEAD. In several cancers, upstream factors in Hippo pathway are inactivated by genetic alterations. When the upstream factors are inactivated, TEAD is activated and forms a complex with YAP/TAZ resulting in enhancement of cell proliferation, drug resistance and so on. In the activation process, S-palmitoylation of TEAD is necessary for binding to YAP/TAZ. Malignant pleural mesothelioma (MPM) is one of cancer types which have genetic alterations in Hippo pathway genes. Although YAP/TAZ-TEAD inhibitor should be an ideal drug for MPM therapy, there are only a few reports about YAP/TAZ-TEAD inhibitor and the efficacy and selectivity are not sufficient. In this study, we succeeded to synthesize a small molecule TEAD inhibitor, K-975, and evaluated its mechanism of action and anti-tumor effect against MPM. Materials/methods: Inhibitory activity of K-975 on YAP/TAZ-TEAD protein-protein interaction (PPI) was evaluated in surface plasmon resonance (SPR) and co-immunoprecipitation assay. The effect of K-975 on palmitoylation status of TEAD was also evaluated. The three-dimensional structure of YAP-binding domain of TEAD1 in complex with K-975 was determined by X-ray crystallography. Anti-tumor effect of K-975 was evaluated by using MPM cell lines. Furthermore, using a derivative of K-975, 2 week-toxicity studies in rats and monkeys were performed. Results: K-975 inhibited YAP-TEAD and TAZ-TEAD PPI in NCI-H226 cells, a human MPM cell line. Also, K-975 inhibited palmitoylation of TEAD. The crystal structure revealed that K-975 directly bound to cysteine residue in YAP-binding domain of TEAD1. This cysteine residue is highly conserved in TEAD family and known as a site of S-palmitoylation. K-975 inhibited the cell proliferation of NCI-H226 with GI50 of about 20 nmol/L. K-975 also induced a change of gene expressions similar to that induced by YAP knockdown. In vivo experiments, K-975 strongly suppressed the tumor growth in several s.c. xenograft models and showed a significant survival benefit in an orthotopic xenograft model. However, 2 week-toxicity studies of a K-975 derivative with optimized bioavailability showed some pathological findings which suggested the renal toxicity. Conclusion: We synthesized a first-in-class drug which directly binds to TEAD protein and inhibits YAP/TAZ-TEAD PPI. K-975 showed a strong anti-tumor effect in pre-clinical MPM models. Although the renal toxicity might cause some difficulty in clinical use, we believe that a K-975 derivative has a possibility to become an effective drug candidate for MPM therapy. Citation Format: Ayumi Kaneda, Toshihiro Seike, Takeshi Uemori, Kensuke Myojo, Kensuke Aida, Tomohiro Danjo, Takahiro Nakajima, Daisuke Yamaguchi, Tomoko Hamada, Yoshiro Tsuji, Kaori Hamaguchi, Mai Yasunaga, Nobumasa Otsubo, Hideyuki Onodera, Yoichi Nishiya, Michihiko Suzuki, Junichi Saito, Toshihiko Ishii, Ryuichiro Nakai. Discovery of a first-in-class TEAD inhibitor which directly inhibits YAP/TAZ-TEAD protein-protein interaction and shows a potent anti-tumor effect in malignant pleural mesothelioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3086.

The YAP/TAZ Pathway in Osteogenesis and Bone Sarcoma Pathogenesis

“…Using fragment-based and computational modeling approaches, the Ω-loop or the α-helix protein interaction domain on YAP, and a hydrophobic palmitoylation pocket on TEAD were identified as targetable domains [233,234,237]. Peptides binding to this pocket may serve as scaffolds for the development of compounds with the potential to break the YAP/TAZ-TEAD interaction [238], and several such compounds are currently in pharmaceutical development [239][240][241][242] (Figure 4). Cells 2020, 9, x 19 of 33 YAP and TAZ are often activated as an alternative survival pathway in drug-resistant cells and can be involved in mechanisms inducing drug resistance [27].…”

Discovery of Covalent Inhibitors Targeting the Transcriptional Enhanced Associate Domain Central Pocket

“… 31 Another covalent TEAD inhibitor ( 4 , K-975 ) showed efficacy in human malignant pleural mesothelioma xenograft models. 32 More recently, a vinylsulfonamide derivative ( 5 , DC-TEADin02 ) was developed via structure-based virtual screening and was shown to covalently bind to TEAD in cells transfected with Flag-TEAD4. 33 Triazole 6 efficiently inhibits TEAD palmitoylation in intestinal epithelium in vivo, 34 and compound 7 was identified via screening a DNA-ecoded library with indole-focused Ugi-peptidomimetics.…”

High Glucose Activates YAP Signaling to Promote Vascular Inflammation

“…Inhibition of the YAP/TAZ signaling pathway was obtained by using either siRNAs against YAP and TAZ or the small molecule TEAD inhibitor K-975 (Kaneda et al, 2020). For siRNAmediated inhibition of YAP/TAZ, HUVECs were transfected with siRNAs using Lipofectamine TM RNAiMAX reagent (Thermo Fischer) in Opti-MEM and L-Glutamine (Gibco) according to the manufacturer's instruction.…”

“…Cells were incubated with transfection complexes at 37 • C in 5% CO2 for 4 h and media was replaced with complete endothelial growth medium for 20 h. Then, HUVEC were cultured for additional 24 h on either normal (5 mM) or high glucose (25 mM) condition in endothelial cell medium added with Growth Medium Supplement Pack (PromoCell). For the purpose of TEAD inhibition, K-975 was synthesized internally based on Kaneda et al (2020). K-975 at 200 nM was added to HUVECs cultured on either normal (5 mM) or high glucose (25 mM) for 24 h, and under either static conditions or subjected to fluidic conditions.…”