(P03-038-24) To Establish a Clinically Suitable Solvent for an Anti-Diabetes Small Molecule BT-141

Objectives: The E4orf1 protein, derived from human adenovirus 36 (Ad36), improves cellular glucose uptake in vitro and in vivo. We previously reported that BT-141, a boron-based small molecule analog of E4orf1 dissolved in Dimethyl sulfoxide (DMSO) improves glycemic control in vitro and in vivo by up-regulating insulin signaling pathway, leading to increased cellular glucose uptake. However, use of DMSO as a clinically safe solvent is not FDA approved. For its potential therapeutic application in humans, we conducted this study to identify FDA approved solvents for BT-141. Moreover, we evaluated the efficacy of BT-141 in presence of these solvents, in activating insulin signaling pathway as determined by AKT-phosphorylation (pAKT).

Methods: 3T3-L1 murine preadipocytes were treated for 2h or 4h with BT-141 (80 µM or 0 µM) dissolved in DMSO, polyethylene glycol (PEG200), or Captisol (a modified cyclodextrin). Whole protein lysates were extracted, concentration determined using the BCA assay and western blot analysis performed to evaluate AKT phosphorylation.

Results: Regardless of the solvent type, BT-141-treated cells showed increased abundance pAKT, indicating activation of the distal insulin signaling pathway, following 2h treatment, which was not observed when cells were treated for 4h. Between the solvents tested, BT-141 dissolved in PEG200 or DMSO showed significantly greater pAKT abundance compared with Captisol.

Conclusions: These findings identify Captisol and PEG200, as two solvents that are FDA approved for human use to further evaluate the anti-diabetes potential of BT-141 in preclinical and clinical trials.

Funding Sources: This research was supported by Insparin Therapeutics.