Polypeptides comprising inteins for engineering outer vesicles

CN122374447APending Publication Date: 2026-07-10EVOX THERAPEUTICS LTD +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
EVOX THERAPEUTICS LTD
Filing Date
2024-09-27
Publication Date
2026-07-10

AI Technical Summary

Technical Problem

In the prior art, premature cleavage of the in-cell peptide between the EV polypeptide or EV localization site and the biotherapy agent leads to reduced cargo loading efficiency and may cause cellular stress and reduced EV yield, making it difficult to purify pure EV products.

Method used

We developed peptides containing ΔI-CM introns and novel exotropic peptide sequences, designed for intron-mediated cleavage with reduced expression in the cytoplasm, and controlled cargo release through pH and temperature-sensitive cleavage to improve EV loading efficiency and bioactivity.

Benefits of technology

It enables more efficient cargo loading and unloading in EVs, reduces the risk of premature cutting, improves the biocompatibility and yield of EVs, and provides a flexible EV loading platform.

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Abstract

The present invention relates to polypeptides comprising inteins (self-cleaving proteins) that are particularly suitable for the production and application of engineered extracellular vesicles (EVs). In particular, the present disclosure relates to fusion proteins comprising newly developed intein variants and EV polypeptides or EV loading moieties that enable more efficient loading of cargo into EVs. Furthermore, the present disclosure relates to polypeptides comprising novel intein variants and newly developed extein sequences that further improve EV loading and / or the biological activity of the produced cargo-loaded EVs. The present invention also relates to polynucleotide constructs encoding these polypeptides and fusion proteins, as well as EVs comprising the polypeptides and fusion proteins in their native and cleaved form.
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