Systems and methods for nucleic acid expression in vivo

Inventors

Debs, Robert James • Heath, Timothy D. • Handumrongkul, Chakkrapong

Assignees

DNARx

Abstract

The present invention provides compositions, systems, kits, and methods for generating expression of one or more proteins and/or biologically active nucleic acid molecules in a subject (e.g., at therapeutic levels for extended periods required to produce therapeutic effects). In certain embodiments, systems and kits are provided that comprise a first composition comprising a first amount of polycationic structures, and a second composition comprising a therapeutically effective amount of expression vectors (e.g., non-viral expression vectors not associated with liposomes) that are CpG-free or CpG-reduced, where the expression vectors comprise a first nucleic acid sequence encoding: i) a first therapeutic protein or proteins, and/or ii) a first biologically active nucleic acid molecule or molecules.

Core Innovation

The invention provides a method of expressing a first therapeutic protein in a subject by administering a first composition intravenously that comprises polycationic structures and is free, or essentially free, of nucleic acid molecules. The method then administers a second composition intravenously within about 300 minutes of administering the first composition, where the second composition comprises CpG-free or CpG-reduced non-viral expression vectors.

The non-viral expression vectors each comprise the same first nucleic acid sequence encoding the same first therapeutic protein, and the ratio of the first amount of the polycationic structures to the non-viral expression vectors is 5:1 to 25:1. As a result of administering the first and second compositions, the first therapeutic protein is expressed in the subject at a level of at least 50 pg/ml in serum for at least 7 consecutive days.

The invention further includes administering, intravenously, a drug agent in the first and/or second composition, or present in a third composition, wherein the drug agent increases or decreases the expression level of the first therapeutic protein and/or the length of time of expression compared to when the drug agent is not administered. In additional embodiments, dexamethasone, dexamethasone palmitate, and/or neutral lipids are administered in the first and/or second composition, or present in a third composition, in combination with the first and second compositions.

Claims Coverage

Two independent claims are provided, centered on a sequential intravenous regimen using a nucleic-acid-free first composition with polycationic structures followed by a CpG-free or CpG-reduced non-viral expression vector second composition within about 300 minutes, and aiming for at least 50 pg/ml in serum for at least 7 consecutive days. The independent claims further differ in whether they explicitly recite dexamethasone, dexamethasone palmitate, and/or neutral lipids within the claim body.

Overall, the independent claims cover a regimen that couples a nucleic-acid-free polycationic first composition with a CpG-free or CpG-reduced non-viral expression vector second composition given within about 300 minutes, using the same therapeutic-protein encoding nucleic acid sequence and a defined polycationic-structure-to-vector ratio to achieve at least 50 pg/ml serum expression for at least 7 consecutive days. Both claims further cover modulation of expression level and/or expression duration using a drug agent, and one independent claim additionally requires co-administration of dexamethasone, dexamethasone palmitate, and/or neutral lipids.

Stated Advantages

Documented Applications