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 A 386 (Cholix 386 ) 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.

What is claimed is: 
     
         1 . A method for delivering a therapeutic cargo to a subject, the method comprising orally administering to the subject a pharmaceutical composition comprising a non-toxic Cholix toxin coupled to the therapeutic cargo. 
     
     
         2 . The method of  claim 1 , wherein the pharmaceutical composition further comprises one or more cleavable linker. 
     
     
         3 . The method of  claim 1 , wherein the pharmaceutical composition is formulated in a capsule or tablet. 
     
     
         4 . The method of  claim 1 , wherein the non-toxic Cholix toxin does not include a domain III. 
     
     
         5 . The method of  claim 1 , wherein the non-toxic Cholix toxin is truncated. 
     
     
         6 . The method of  claim 1 , wherein the non-toxic Cholix toxin is truncated at amino acid residue Ala 386 . 
     
     
         7 . The method of  claim 1 , wherein the non-toxic Cholix toxin does not have a functional domain III. 
     
     
         8 . The method of  claim 1 , wherein the non-toxic Cholix toxin has a mutation in domain III. 
     
     
         9 . The method of  claim 1 , wherein the non-toxic Cholix toxin has a mutation at amino acid residue Glu 581 . 
     
     
         10 . The method of  claim 1 , wherein the therapeutic cargo is a medicament, a macromolecule, a small molecule, a nanoparticle, or a dendrimer-based carrier. 
     
     
         11 . The method of  claim 1  wherein the therapeutic cargo is a medicament selected from the list consisting of antineoplastic compound, methylhydrazines, steroid hormones, immunosuppressives, immunostimulants, antimicrobial compounds, antifungals, antivirals, blood clotting proteins, antihalmintics, radiopharmaceutics, gastrointestinal drugs, hematologic compounds, immunoglobulins, anticoagulants, fibrolysin inhibitors, peripheral anti-adrenergic drugs, centrally acting antihypertensive drugs, antihypertensive direct vasodilators, drugs affecting renin-angiotensin system, calcium entry blockers, neuromuscular blocking drugs, centrally acting muscle relaxants, neurotransmitters and neurotransmitter agents, diuretic drugs, and antimigraine drugs. 
     
     
         12 . The method of  claim 1 , wherein the therapeutic cargo is a macromolecule or small molecule selected from the group consisting of siRNA, PNA, miRNA, DNA, plasmid and antisense molecules. 
     
     
         13 . The method of  claim 1 , wherein the therapeutic cargo is delivered at a dose from about 1 μg to about 1 g. 
     
     
         14 . The method of  claim 1 , wherein the therapeutic cargo is delivered at a dose from about 10 μg to about 100 mg. 
     
     
         15 . The method of  claim 1 , wherein the therapeutic cargo is delivered at a dose from about 10 μg to about 1000 μg.