Cell membrane translocation of regulated snare inhibitors, compositions therefor, and methods for treatment of disease

Abstract

Compositions and methods of modulating cellular function and treatment of disease in mammals comprising locally administering a regulated SNARE inhibitor and a translocating agent to the mammal. Regulated SNARE inhibitors include clostridial neurotoxins, tetanus neurotoxin and their free light chain portions and IgA protease. Translocating agents include acids, encapsulating vectors, and transduction domains.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of International Application No. PCT / US04 / 05436, filed Feb. 24, 2004, entitled Cell Membrane Translocation of Regulated Snare Inhibitors, Compositions Therefor, and Methods for Treatment of Disease; and U.S. Provisional Application No. 60 / 449,107 filed Feb. 24, 2003, entitled Compositions of Amphipathic Pharmaceuticals and Methods For Their Use, by I. Sanders, which applications are hereby incorporated herein by reference in their entirety.FIELD OF THE INVENTION [0002] This invention relates to translocation of bioactive molecules through cell membranes. More specifically, the invention relates to cell membrane translocation of regulated SNARE inhibitors, compositions and formulations thereof, and methods for treatment of disease. BACKGROUND OF THE INVENTION [0003] A promising area of pharmaceutical intervention is the use of macromolecules that act within cells. These include gene therapy, natural an...

AI Technical Summary

Benefits of technology

[0037] The bioactive substance for use in the invention can be any substance or molecule that induces a biological response in mammalian cells or a therapeutic effect in a mammal. Preferably, the bioactive substance is a bioactive part of a natural toxin. More preferably, the bioactive substance is a regulated SNARE inhibitor, most preferably, the bioactive substance is the light chain of a clostridial neurotoxin, preferably, the free light chain or its analogue, a protein called IgA protease.

Problems solved by technology

This route is difficult for most complex pharmaceuticals as they are degraded during the prolonged endosomal stage.

However, few amphipathic protein conjugates are presently approved for clinical use.

Second, they can block neuromuscular transmission for extended periods, from days to months depending on the serotype.

Third, in most clinical applications they have been used at doses that are below the level of immunological recognition.

While passing through the membrane the disulfide bond is broken and the light chain is released into the cytoplasm and exerts its toxic effect.

These high doses risk local and systemic side effects.

But the A-B fragment can cross the cell membrane and paralyze the neuromuscular synapse and cause a flaccid paralysis.

Unfortunately, however, as discussed below, introduction of these novel compounds is limited because of the inefficiencies of the endosomic process.

Although, as discussed above, amphipathic protein conjugates—such as clostridial neurotoxins—have potential medical applications, their usefulness is limited by inefficient transport into cells.

This is a disadvantage for a variety of reasons.

This inefficiency increases the incidence of side effects and the induction of immune reactions.

Accordingly, translocation across the endosome membrane is the rate-limiting step in the entry of the active moiety into the cell.

Another disadvantage of amphipathic protein conjugates, is that the endosome contains its own assortment of enzymes that can degrade the conjugate.