Amino acid sequences capable of facilitating penetration across a biological barrier

Abstract

This invention relates to novel pharmaceutical penetration compositions capable of facilitating penetration of at least one effector across biological barriers. The invention also relates to methods of treating or preventing diseases by administering penetration compositions to affected subjects.

1 . A composition for non-invasive translocation of at least one effector across a biological barrier, said composition comprising: 
 (a) a therapeutically effective amount of said at least one effector;    (b) a counter ion to the at least one effector; and    (c) an  Escherichia coli  related penetrating peptide    wherein said penetrating peptide is hydrophobized.    
     
     
         2 . The composition of  claim 1 , wherein the penetrating peptide comprises an amino acid sequence selected from the group consisting of: SEQ ID NOS: 3, 25, 26, and 27.  
     
     
         3 . The composition of  claim 1  further comprising a pharmaceutically acceptable carrier, diluent, or excipient, or a combination thereof.  
     
     
         4 . The composition of  claim 3 , wherein said composition is in the form of a capsule or tablet.  
     
     
         5 . The composition of  claim 4 , wherein said capsule or tablet is enteric-coated.  
     
     
         6 . The composition of  claim 3 , wherein said composition is in the form selected from the group consisting of an aqueous dispersion, a suspension, and an emulsion.  
     
     
         7 . The composition of  claim 3 , wherein said composition is in the form of a cream.  
     
     
         8 . The composition of  claim 3 , wherein said composition is in the form of an ointment.  
     
     
         9 . The composition of  claim 3 , wherein said composition is in the form of a suppository.  
     
     
         10 . The composition of  claim 1 , wherein said at least one effector is an impermeable molecule.  
     
     
         11 . The composition of  claim 10 , wherein said impermeable molecule is cationic.  
     
     
         12 . The composition of  claim 10 , wherein said impermeable molecule is anionic.  
     
     
         13 . The composition of  claim 12 , wherein said anionic impermeable molecule is a polysaccharide.  
     
     
         14 . The composition of  claim 13 , wherein said polysaccharide is a glycosaminoglycan.  
     
     
         15 . The composition of  claim 14 , wherein said glycosaminoglycan is selected from the group consisting of: heparin; heparan sulfate; chondroitin sulfate; dermatan sulfate; hyaluronic acid and pharmaceutically acceptable salts thereof.  
     
     
         16 . The composition of  claim 1 , wherein said at least one effector is a pharmaceutically active agent selected from the group consisting of: a hormone, a growth factor, a neurotrophic factor, an anticoagulant, a bioactive molecule, a toxin, an antibiotic, an anti-fungal agent, an antipathogenic agent, an antigen, an antibody, an antibody fragment, an immunomodulator, a vitamin, an antineoplastic agent, an enzyme, and a therapeutic agent.  
     
     
         17 . The composition of  claim 16 , wherein said bioactive molecule is selected from the group consisting of: insulin; erythropoietin (EPO); glucagon-like peptide 1 (GLP-1); αMSH; parathyroid hormone (PTH); growth hormone; calcitonin; interleukin-2 (IL-2); α1-antitrypsin; granulocyte/monocyte colony stimulating factor (GM-CSF); granulocyte colony stimulating factor (G-CSF); T20; anti-TNF antibodies; interferon α; interferon β; interferon γ; lutenizing hormone (LH); follicle-stimulating hormone (FSH); enkephalin; dalargin; kyotorphin; basic fibroblast growth factor (bFGF); hirudin; hirulog; lutenizing hormone releasing hormone (LHRH) analog; brain-derived natriuretic peptide (BNP); and neurotrophic factors.  
     
     
         18 . The composition of  claim 16 , wherein said anti-fungal agent is Caspofungin.  
     
     
         19 . The composition of  claim 16 , wherein said vitamin is vitamin B12.  
     
     
         20 . The composition of  claim 16 , wherein said antibiotic is an aminoglycoside antibiotic.  
     
     
         21 . The composition of  claim 20 , wherein said aminoglycoside antibiotic is selected from the group consisting of Gentamycin, Amikacin, Tobramycin, and Neomycin.  
     
     
         22 . The composition of  claim 1 , wherein said effector is a nucleic acid or a nucleic acid mimetic.  
     
     
         23 . The composition of  claim 1 , wherein said counter ion is an amphipathic molecule.  
     
     
         24 . The composition of  claim 23 , wherein said amphipathic molecule is cationic.  
     
     
         25 . The composition of  claim 23 , wherein said amphipathic molecule is anionic.  
     
     
         26 . The composition of  claim 25 , wherein said anionic amphipathic molecule is derived from a strong acid selected from the group consisting of sulfonate and phosphonate, and wherein said anionic amphipathic molecule further comprises a hydrophobic moiety.  
     
     
         27 . The composition of  claim 25 , wherein the anionic amphipathic molecule is selected from the group consisting of: sodium dodecyl sulphate and dioctyl sulfosuccinate.  
     
     
         28 . The composition of  claim 24 , wherein said cationic amphipathic molecule is a quaternary amine comprising a hydrophobic moiety.  
     
     
         29 . The composition of  claim 28 , wherein said quaternary amine has the general structure of:  
       
         
           
           
               
               
           
         
       
       wherein R1, R2, R3 and R4 are alkyl or aryl residues.  
     
     
         30 . The composition of  claim 29 , wherein said quaternary amine is a benzalkonium derivative.  
     
     
         31 . The composition of  claim 1 , wherein said counter ion is an ionic liquid forming cation.  
     
     
         32 . The composition of  claim 31 , wherein said ionic liquid forming cation is selected from the group consisting of: imidazolium derivatives; pyridinium derivatives; phosphonium compounds; and tetralkylammonium compounds.  
     
     
         33 . The composition of  claim 32 , wherein said imidazolium derivative has the general structure of 1-R1-3-R2-imidazolium, wherein R1 and R2 are linear or branched alkyls with 1 to 12 carbons.  
     
     
         34 . The composition of  claim 33 , wherein said imidazolium derivatives further comprise a halogen or an alkyl group substitution.  
     
     
         35 . The composition of  claim 33 , wherein said imidazolium derivative is selected from the group consisting of: 1-ethyl-3-methylimidazolium; 1-butyl-3 methylimidazolium; 1-hexyl-3-methylimidazolium; 1-methyl-3-octylimidazolium; 1-methyl-3-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoroctyl)-imidazolium; 1,3-dimethylimidazolium; and 1,2-dimethyl-3-propylimidazolium.  
     
     
         36 . The composition of  claim 32 , wherein said pyridinium derivative has the general structure of 1-R1-3-R2-pyridinium, wherein R1 is a linear or branched alkyl with 1 to 12 carbons, and R2 is H or a linear or branched alkyl with 1 to 12 carbons.  
     
     
         37 . The composition of  claim 36 , wherein said pyridinium derivatives further comprise a halogen or an alkyl group substitution.  
     
     
         38 . The composition of  claim 36 , wherein said pyridinium derivative is selected from the group consisting of 3-methyl-1-propylpyridinium, 1-butyl-3-methylpyridinium, and 1-butyl-4-methylpyridinium.  
     
     
         39 . The composition of  claim 1 , further comprising a polyanionic molecule.  
     
     
         40 . The composition of  claim 39 , wherein said polyanionic molecule is phytic acid.  
     
     
         41 . The composition of  claim 1 , further comprising a surface active agent.  
     
     
         42 . The composition of  claim 41 , wherein said surface active agent is selected from the group consisting of a poloxamer, Solutol HS15, Cremophore, and bile acids.  
     
     
         43 . The composition of  claim 1 , wherein said composition is dissolved in an at least partially water soluble solvent.  
     
     
         44 . The composition of  claim 43 , wherein said at least partially water soluble solvent is selected from the group consisting of: n-butanol; isoamyl (=isopentyl) alchohol; iso-butanol; iso-propanol; propanol; ethanol; ter-butanol alcohols; polyols; DMF; DMSO; ethers; amides; esters; and mixtures thereof.  
     
     
         45 . The composition of  claim 1 , wherein any one or more of the components of the composition is lyophilized.  
     
     
         46 . The composition of  claim 1 , wherein said composition further comprises a hydrophobic carrier comprising at least one hydrophobic molecules, wherein said molecules are aliphatic, aromatic, or mixtures thereof.  
     
     
         47 . The composition of  claim 46 , wherein said aliphatic hydrophobic molecules are selected from the group consisting of fatty acids, monoglycerides, diglycerides, triglycerides, ethers, and cholesterol esters of fatty acids.  
     
     
         48 . The composition of  claim 47 , wherein said triglyceride is tricaprin.  
     
     
         49 . The composition of  claim 46 , wherein said aromatic hydrophobic molecule is benzyl benzoate.  
     
     
         50 . The composition of  claim 1 , further comprising at least one protective agent.  
     
     
         51 . The composition of  claim 50 , wherein said protective agent is a protease inhibitor selected from the group consisting of: aprotinin; Bowman-Birk inhibitor; soybean trypsin inhibitor; chicken ovomucoid; chicken ovoinhibitor; human pancreatic trypsin inhibitor; camostate mesilate; flavonoid inhibitors; antipain; leupeptin; paminobenzamidine; AEBSF; TLCK; APMSF; DFP; PMSF; poly(acrylate) derivatives; chymostatin; benzyloxycarbonyl-Pro-Phe-CHO; FK-448; sugar biphenylboronic acids complexes; β-phenylpropionate; elastatinal; methoxysuccinyl-Ala-Ala-Pro-Valchloromethylketone (MPCMK); EDTA; chitosan-EDTA conjugates; amino acids; dipeptides; tripeptides; amastatin; bestatin; puromycin; bacitracin; phosphinic acid dipeptide analogues; α-aminoboronic acid derivatives; Na-glycocholate; 1,10-phenantroline; acivicin; L-serine-borate; thiorphan; and phosphoramidon.  
     
     
         52 . The composition of  claim 1 , wherein the penetrating peptide is further modified, via one or more peptidic bonds, to enable protection from gastrointestinal proteolysis.  
     
     
         53 . The composition of  claim 1 , wherein said penetrating peptide further contains lysine residues, interspaced by glycine, alanine or serine residues, added at the C-terminus of the penetrating peptide, and wherein the free amino groups of said lysine residues are acylated.  
     
     
         54 . The composition of  claim 53 , wherein acylation utilizes long-chain fatty acids selected from the group of: stearoyl, palmitoyl, oleyl, ricinoleyl, lauroyl and myristoyl.  
     
     
         55 . The composition of  claim 3 , wherein the composition further comprises two or more of the following: a non-ionic detergent, an ionic detergent, a protease inhibitor, and a reducing agent.  
     
     
         56 . The composition of  claim 55 , wherein the non-ionic detergent is a poloxamer or polyehtyleneglycol-15-hydroxystearate (Solutol HS15).  
     
     
         57 . The composition of  claim 56 , wherein the poloxamer is polyoxyethylene-polyoxypropylene block copolymer.  
     
     
         58 . The composition of  claim 55 , wherein the ionic detergent is a bile salt.  
     
     
         59 . The composition of  claim 58 , wherein the bile salt is Taurodeoxycholate.  
     
     
         60 . The composition of  claim 55 , wherein the protease inhibitor is selected from the group consisting of: aprotinin; Bowman-Birk inhibitor; soybean trypsin inhibitor; chicken ovomucoid; chicken ovoinhibitor; human pancreatic trypsin inhibitor; camostate mesilate; flavonoid inhibitors; antipain; leupeptin; p-aminobenzamidine; AEBSF; TLCK; APMSF; DFP; PMSF; poly(acrylate) derivatives; chymostatin; benzyloxycarbonyl-Pro-Phe-CHO; FK-448; sugar biphenylboronic acids complexes; β-phenylpropionate; elastatinal; methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone (MPCMK); EDTA; chitosan-EDTA conjugates; amino acids; dipeptides; tripeptides; amastatin; bestatin; puromycin; bacitracin; phosphinic acid dipeptide analogues; α-aminoboronic acid derivatives; Na-glycocholate; 1,10-phenantroline; acivicin; L-serine-borate; thiorphan; and phosphoramidon.  
     
     
         61 . The composition of  claim 55 , wherein the reducing agent is N-acetyl-L-cystein (NAC).  
     
     
         62 . A kit for treating or preventing a disease or pathological condition comprising, in one or more containers, a therapeutically or prophylactically effective amount of the composition of  claim 3 .  
     
     
         63 . The composition of  claim 1 , wherein said penetrating peptide further comprises a chemical modification.  
     
     
         64 . The composition of  claim 63 , wherein the chemical modification comprises the attachment of one or more polyethylene glycol residues to the penetrating peptide.  
     
     
         65 . A method of mucosal vaccination, the method comprising administering to a subject in need of vaccination the composition of  claim 3 , wherein the at least one effector comprises an antigen to which vaccination is desired.  
     
     
         66 . The method of  claim 65 , wherein the antigen to which vaccination is desired is selected from the group consisting of PA for use in a vaccine against Anthrax, and HBs for use in a vaccine against Hepatitis B.  
     
     
         67 . A method of treating or preventing a disease or pathological condition, said method comprising administering to a subject in which such treatment or prevention is desired, the composition of  claim 3 , in an amount sufficient to treat or prevent said disease or said pathological condition in said subject.  
     
     
         68 . The method of  claim 67 , wherein said disease or said pathological condition is selected from the group consisting of: endocrine disorders; diabetes; infertility; hormone deficiencies; osteoporosis; neurodegenerative disorders; Alzheimer's disease; dementia; Parkinson's disease; multiple sclerosis; Huntington's disease; cardiovascular disorders; atherosclerosis; hyper- and hypocoagulable states; coronary disease; cerebrovascular events; metabolic disorders; obesity; vitamin deficiencies; renal disorders; renal failure; haematological disorders; anemia of different entities; immunologic and rheumatologic disorders; autoimmune diseases; immune deficiencies; infectious diseases; viral infections; bacterial infections; fungal infections; parasitic infections; neoplastic diseases; multi-factorial disorders; impotence; chronic pain; depression; different fibrosis states; and short stature.