Discovery of TDI-10229: A Potent and Orally Bioavailable Inhibitor of Soluble Adenylyl Cyclase (sAC, ADCY10)

Abstract

Soluble adenylyl cyclase (sAC) has gained attention as a potential therapeutic target given the role of this enzyme in intracellular signaling. We describe successful efforts to design improved sAC inhibitors amenable for in vivo interrogation of sAC inhibition to assess its potential therapeutic applications. This work culminated in the identification of TDI-10229 (12), which displays nanomolar inhibition of sAC in both biochemical and cellular assays and exhibits mouse pharmacokinetic properties sufficient to warrant its use as an in vivo tool compound.

Assessing potency and binding kinetics of soluble adenylyl cyclase (sAC) inhibitors to maximize therapeutic potential

“…Biochemical potency and jump dilution assays used N-terminal tagged GST-sAC t (Litvin et al, 2003); SPR used highly pure C-terminal tagged sAC t -His6 that was prepared via sequential chromatography that included, in this order, affinity (Ni-Sepharose HP from GE Healthcare), ion exchange (Mono Q from GE Healthcare), and size exclusion (Superdex 200 from GE Healthcare) chromatographies. The synthesis of TDI-10229 is described in Fushimi et al, 2021 and the synthesis of other compounds is described in Miller et al, 2022. In vitro adenylyl cyclase activity assay with purified sAC protein (biochemical potency assay)…”

Transforming Academic Drug Discovery

“…As one such instance, a TDI drug discovery program focused on the identification of potent, soluble adenylyl cyclase (sAC, ADCY10) inhibitors suitable for the treatment of hypotony, [22] a rare, blinding condition characterized by low intraocular pressure. Conversion of a weakly active screening lead, LRE1, [23] yielded the orally active, highly selective, and potent sAC inhibitor TDI‐10229 [24] . The availability of TDI‐10229 opened a path for an effective oral contraceptive strategy for episodic contraceptive use; such a pill would provide on demand male (or female) reversible contraception [25] …”

“…Conversion of a weakly active screening lead, LRE1, [23] yielded the orally active, highly selective, and potent sAC inhibitor TDI-10229. [24] The availability of TDI-10229 opened a path for an effective oral contraceptive strategy for episodic contraceptive use; such a pill would provide on demand male (or female) reversible contraception. [25]…”

Strategies to safely target widely expressed soluble adenylyl cyclase for contraception

“…We took advantage of the drug design expertise of a unique public-private partnership ( Meinke, 2022 ) to improve the potency, selectivity, and drug-like characteristics of LRE1. Ultimately, these efforts increased potency for sAC over 10,000-fold with corresponding increased efficacy in cell-based assays, diminished cross-reactivity with other mammalian nucleotidyl cyclases, and no significant cytotoxicity ( Fushimi et al, 2021 ).…”

“…In C1-C2 sAC isoforms, the C1 domain contributes key catalytic residues that would be absent if sAC-C2 isoforms were to homodimerize, and no known C1-like binding partner has yet been identified for sAC-C2. The newest generation of potent and selective inhibitors were generated via structure-based drug design using a crystal structure of human C1-C2 containing sAC ( Fushimi et al, 2021 ). These more potent inhibitors, which fill both the BBS and the active site formed at the interface of C1 and C2, making contacts in both domains, were used to validate sAC as a contraceptive target.…”