De Novo Design of Peptidic Positive Allosteric Modulators Targeting TRPV1 with Analgesic Effects

Transient receptor potential vanilloid 1 (TRPV1) ion channel is a nociceptor critically involved in pain sensation. Direct blockade of TRPV1 exhibits significant analgesic effects but also incurs severe side effects such as hyperthermia, causing failures of TRPV1 inhibitors in clinical trials. In order to selectively target TRPV1 channels that are actively involved in pain‐sensing, peptidic positive allosteric modulators (PAMs) based on the high‐resolution structure of the TRPV1 intracellular ankyrin‐repeat like domain are de novo designed. The hotspot centric approach is optimized for protein design; its usage in Rosetta increases the success rate in protein binder design. It is demonstrated experimentally, with a combination of fluorescence resonance energy transfer (FRET) imaging, surface plasmon resonance, and patch‐clamp recording, that the designed PAMs bind to TRPV1 with nanomolar affinity and allosterically enhance its response to ligand activation as it is designed. It is further demonstrated that the designed PAM exhibits long‐lasting in vivo analgesic effects in rats without changing their body temperature, suggesting that they have potentials for developing into novel analgesics. Transient receptor potential vanilloid 1 (TRPV1) ion channel is critically involved in pain sensation. Positive allosteric modulators (PAMs) of TRPV1 are de novo designed based on the structure of TRPV1. Surface plasmon resonance and patch‐clamp recording demonstrate that the PAMs bind to TRPV1 and allosterically enhance its activities. The PAM exhibits analgesic effects without changing body temperature in rats.