Expression constructs, fusogenic lipid-based nanoparticles and methods of use thereof

Inventors

Scholz, Matthew Rein

Assignees

Oisin Biotechnologies Inc

Abstract

Provided nucleic acid-based expression construct for the target cell-specific production of a therapeutic protein, such as a pro-apoptotic protein, within a target cell, including a target cell that is associated with aging, disease, or other condition, in particular a target cell that is a senescent cell or a cancer cell. Also provided are formulations and systems, including fusogenic lipid nanoparticle (LNP) formulations and systems, for the delivery of nucleic acid-based expression constructs as well as methods for making and using such nucleic acid-based expression constructs, formulations, and systems for reducing, preventing, and/or eliminating the growth and/or survival of a cell, such as a senescent cell and/or a cancer cell, which is associated with aging, disease, or other condition as well as methods for the treatment of aging, disease, or other conditions by the in vivo administration of a formulation, such as a fusogenic LPN formulation, comprising an expression construct for the target cell-specific production of a therapeutic protein, such as a pro-apoptotic protein, in a target cell that is associated with aging, disease, or other condition, in particular a target cell that is a senescent cell or a cancer cell.

Core Innovation

The invention provides an expression construct for production of a therapeutic protein within a target cell. The expression construct includes a p53 transcriptional promoter and a heterologous nucleic acid under regulatory control of the p53 transcriptional promoter, and the heterologous nucleic acid encodes a therapeutic protein that reduces growth, survival, or a combination thereof, of the target cell.

In the disclosed implementations, the therapeutic protein is selected among pro-apoptotic or cytotoxic proteins, including CASP3, CASP8, and CASP9, with specific reference to inducible CASP9 and non-inducible CASP9 variants such as iCasp9 and saCasp9. The therapeutic protein is produced in target cells based on promoter control associated with p53 transcriptional regulation.

The disclosed system further combines the expression construct with fusogenic lipid nanoparticles for delivery and target-cell-specific production of the therapeutic protein. The document also describes optional features including antigen or immunostimulatory cargo such as CD40L, marker encoding for imaging or reporting purposes, and safety features such as tamoxifen-inducible Cre. The delivery approach includes cytoplasmic delivery that bypasses endocytosis and is supported by in vitro and in vivo observations of targeting and apoptosis-related outcomes.

Claims Coverage

The independent claim set includes one independent claim focused on an expression construct with a p53 transcriptional promoter controlling a therapeutic-protein-encoding heterologous nucleic acid, producing a therapeutic protein within a target cell to reduce growth and/or survival. Additional inventive features are provided by refinements in dependent claims for specific therapeutic proteins and optional nanoparticle-based delivery and targeting parameters.

Overall, the disclosed claim coverage centers on p53 transcriptional promoter control of a heterologous nucleic acid encoding a therapeutic protein that reduces growth and/or survival in target cells, with further refinements for inducible vs non-inducible CASP9, optional CD40L immunostimulatory cargo, and fusogenic LNP delivery features including a p14 FAST identity constraint and specified sets of cancer-cell targets.

Stated Advantages

Documented Applications