Fusogenic lipid nanoparticles for target cell-specific production of a therapeutic protein

Inventors

Scholz, Matthew Rein • Lewis, John David • Hudson, Gary Charles

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 disclosed invention provides a lipid-based nanoparticle (LNP) formulation for targeted production of a therapeutic protein within target cells. The lipid nanoparticle vector comprises a fusogenic protein, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) at a molar ratio of 22.5–37.5 mole %, and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG-PEG) at a molar ratio of 3–5 mole % for non-specific delivery of an expression construct to mammalian cells that comprise both target cells and non-target cells.

The formulation further includes an expression construct configured for preferential production of the therapeutic protein within the target cells. The expression construct comprises a transcriptional promoter activated in response to one or more factors preferentially produced within the target cells as compared to the non-target cells, and a nucleic acid encoding the therapeutic protein is operably linked to and under regulatory control of the transcriptional promoter, so that the therapeutic protein is produced within the target cells but is not produced in the non-target cells.

The disclosed systems are also directed to treating a condition by administering lipid-based nanoparticle formulations that deliver multiple nucleic acid sequences under different transcriptional promoter controls. In particular, p16 and p53 transcriptional promoters operably control first and second polynucleotides encoding first and second therapeutic proteins, respectively, with the therapeutic proteins selected from apoptosis-/caspase-related proteins, including caspases, inducible caspases, self-activating caspases, and other pro-death enzymes.

Claims Coverage

Two independent claims are identified, covering a fusogenic LNP formulation for preferential target-cell-specific production of a therapeutic protein and a treatment method using LNP formulations that deliver two promoter-controlled nucleic acid sequences. Across these claims, the inventive coverage centers on fusogenic LNP delivery plus transcriptional promoter activation by target-cell-preferential factors, and on promoter-defined apoptosis/caspase therapeutic proteins.

The claim set covers a fusogenic LNP vector with specified DOPE and DMG-PEG composition delivering an expression construct whose transcriptional promoter is activated by factors preferentially present in target cells, producing a therapeutic protein that reduces, prevents, or eliminates growth or survival in target cells while not being produced in non-target cells. It further covers treating a condition using LNP formulations that include two nucleic acid sequences controlled by p16 and p53 transcriptional promoters to drive two therapeutic proteins.

Stated Advantages

Documented Applications