Systems and methods of delivery of bioactive agents using bacterial toxin-derived transport sequences

Abstract

The field of the present invention relates, in part, to a strategy for novel pharmaceutical applications. More specifically, the present invention relates to a non-toxic mutant form of the Vibrio cholera Cholix gene (ntCholix), a variant of Cholix truncated at amino acid A386 (Cholix386) and the use of other various Cholix-derived polypeptide sequences to enhance intestinal delivery of biologically-active therapeutics. Importantly, the systems and methods described herein provide for the following: the ability to deliver macromolecule doses without injections; the ability to deliver cargo, such as (but not limited to) siRNA or antisense molecules into intracellular compartments where their activity is required; and the delivery of nanoparticles and dendrimer-based carriers across biological membranes, which otherwise would have been impeded due to the barrier properties of most such membranes.

Description

RELATED PATENT APPLICATIONS[0001]This application is a Continuation of U.S. Ser. No. 13 / 822,435, filed Mar. 12, 2013, which claims priority to and benefit of U.S. Provisional Application No. 61 / 403,394, filed on Sep. 15, 2010, each of which is incorporated in its entirety by reference herein.TECHNICAL FIELD[0002]The field of the present invention relates, in part, to a strategy for novel pharmaceutical applications. More specifically, the present invention relates to a non-toxic mutant form of the Vibrio cholera Cholix gene (ntCholix), a variant of Cholix truncated at amino acid A386 (Cholix386) and the use of other various Cholix-derived polypeptide sequences to enhance intestinal delivery of biologically-active therapeutics. Importantly, the systems and methods described herein provide for the following: the ability to deliver macromolecule doses without injections; the ability to deliver cargo, such as (but not limited to) siRNA or antisense molecules into intracellular compartme...

AI Technical Summary

Benefits of technology

The present invention is about a protein called Cholix that can efficiently move across polarized cells in the airways and intestine. This discovery suggests that certain parts of the Cholix protein can be used to transport proteins and nanoparticles across membranes, which is useful for developing new pharmaceutical applications.

Problems solved by technology

Unfortunately, however, the efficient transport across this simple, single layer of cells remains a substantial barrier due to the intracellular trafficking to destructive lysosome compartments after endosomal uptake of polypeptides at the luminal surface; Woodley, J. F., Crit Rev Ther Drug Carrier Syst, 11(2-3):p.

While transient opening and closing of TJ structures can facilitate transport of peptides across intestinal epithelia, this approach has key limitations: e.g., it does not work well for molecules above ˜5 kDa; it has the potential for non-selective entry of materials into the body from the intestinal lumen; and it represents a route that involves only a small fraction of the surface area of the intestinal epithelium.

Searching the genome of V. cholera for PE-like nucleotide sequences fails to result in a match of any kind.

Nevertheless, Cholix and PE appear to have distinct cell receptor interactions, demonstrating clear differences that are sufficient to suggest very different and unanticipated applications for both oral biologics and the intracellular delivery of bioactive agents.

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