# Bis-prodrug cryopreserved lipid nanoparticles with enzymatically triggered release

## TL;DR

Researchers developed a stable lipid nanoparticle formulation that can preserve and release a water-soluble drug in response to enzymes after freeze-drying.

## Contribution

A novel bis-prodrug LNP formulation with cryopreservability and enzyme-triggered drug release is introduced.

## Key findings

- 50/50 Brij/lipoid surfactant ratios provided optimal redispersibility after freeze-drying.
- The 3:1 prodrug-to-tricaprin ratio achieved stable particle sizes below 150 nm.
- Esterase-mediated drug release was sustained for up to 9 weeks.

## Abstract

Lipid nanoparticle (LNP) formulations have emerged as a versatile platform for the delivery of therapeutics. However, achieving long-term stability and effective delivery of water-soluble small molecule drugs remains a challenge. In this study, we demonstrate a cryopreservable LNP formulation incorporating a hydrophobically modified bis-prodrug of lamivudine. By systematically varying the surfactant composition by combining a PEGylated surfactant (Brij S20) with an unPEGylated, zwitterionic lipid (Lipoid S100), we identify formulations that maintain colloidal stability following freeze–drying and redispersion in the presence of 10% w/v sucrose. Particle size measurements before and after lyophilisation indicate that surfactant ratio significantly impacts redispersibility, with 50/50 Brij/lipoid compositions offering the best performance. A core composition comprising the prodrug and tricaprin at either 1 : 1 or 3 : 1 ratio was evaluated, with the 3 : 1 formulation achieving redispersed particle sizes below 150 nm and low polydispersity. Enzymatic studies using porcine liver esterase confirm slow, sustained conversion of the bis-prodrug to active lamivudine over up to 9 weeks. This work highlights the opportunity of a prodrug-based strategy to formulate water-soluble APIs into stable, freeze-dried LNPs, enabling controlled, enzyme-responsive release. These findings offer insight into how surfactant composition influences freeze–drying compatibility and provide a platform for the development of LNP systems for small molecule delivery.

Bis-prodrug-loaded LNPs show robust cryostability and prolonged esterase-mediated drug release after lyophilisation and rehydration.