Anionic-core composition for delivery of therapeutic agents, and methods of making and using the same

Abstract

The present invention is directed to compositions comprising a polymer backbone with protective chain and anionic groups, and a cationic therapeutic agent. The present invention is directed to compositions for treating infections, inflammatory diseases, excess growth, and damaged cells and organs.

1 - 75 . (canceled) 
     
     
         76 . A pharmaceutical composition, comprising:
 (a) a pharmaceutically acceptable excipient;   (b) a polymer comprising:
 (i) a polymer backbone selected from polylysine, polyornithine, polyarginine, polyglutamate, polyaspartate, polycysteine, polyserine, polythreonine, polytyrosine, chondroitin sulfate, dermatan sulfate, keratan sulfate, carageenan, pectin, fucoidan, dextran, polymethylacrylic acid, polyacrylic acid, and polyallylamine; 
 (ii) multiple protective chains covalently linked to the polymer backbone, wherein each protective chain is independently selected from a group consisting of a linear poly(ethyleneglycol) and a linear methoxy poly(ethyleneglycol), and each protective chain is singly terminally covalently linked to a monomeric unit on the polymer backbone via one covalent bond; 
 (iii) an anionic group attached to a monomeric unit on the polymer backbone by a covalent bond, wherein the anionic group is selected from the group consisting of phosphate, sulfate, sulfonate, and carboxyl, or 
 an anionic cluster comprising two or more anionic groups attached to a monomeric unit on the polymer backbone by a single covalent bond, wherein the anionic groups are each selected from the group consisting of phosphate, sulfate, sulfonate, and carboxyl; and 
   (c) a load molecule linked directly by electrostatic interaction to the anionic group or the anionic cluster of the polymer without an intermediary metal ion, wherein the load molecule:
 (i) comprises a cell penetrating peptide; 
 (ii) comprises an anionic binding domain; or 
 (iii) has an isoelectric point greater than 7.3, 
   wherein at least 86.7% of the load molecule remains linked to the anionic group or the anionic cluster in the presence of 100 mM aqueous NaCl solution.   
     
     
         77 . The composition of  claim 76 , wherein the load molecule is selected from the group consisting of: (i) a peptide; (ii) a protein; and (iii) a small organic molecule. 
     
     
         78 . The composition of  claim 76 , wherein the load molecule is a peptide or a protein comprising a cell penetrating contiguous sequence of 5-10 amino acids, and wherein the number of basic amino acid (lysine and arginine) minus the number of acidic amino acids (glutamate and aspartate) is 2 or greater. 
     
     
         79 . The composition of  claim 78 , wherein the cell penetrating contiguous sequence in the peptide comprises of a sequence selected from the group consisting of: 1) Lys-Lys-Lys-Lys, 2) Lys-Lys-Lys-Arg, 3) Lys-Lys-Arg-Lys, 4) Lys-Lys-Arg-Arg, 4) Lys Arg-Arg-Lys, 5) Lys-Arg-Arg-Arg, and 6) Arg-Arg-Arg-Arg, wherein any amino acid can be D or L isomer and the orientation of the sequence as presented can be amino to carboxyl or carboxyl to amino. 
     
     
         80 . The composition of  claim 76 , wherein the load molecule comprises an agent selected from the group consisting of SEQ ID NOS: 1-59, lysostaphin, an epithelial growth factor (EGF), a heparin binding EGF, an interferon, a Fibroblast growth factor (FGF), a Vascular endothelial growth factor (VEGF), Hepatocyte growth factor (HGF), a Transforming growth factor (TGF), Nerve growth factor (NGF), and a Platelet derived growth factor (PDGF). 
     
     
         81 . The composition of  claim 76 , wherein the load molecule wherein the load molecule comprises an anti-infective agent selected from the group consisting of lysostaphin, interferon, and SEQ ID NOS: 12-45. 
     
     
         82 . The composition of  claim 76 , wherein the load molecule comprises an agent selected from the group consisting of the last 11 amino acids of SEQ ID NO: 1, the last 8 amino acids of SEQ ID NO: 5, the last 12 amino acids of SEQ ID NO: 7, the last 10 amino acids of SEQ ID NO: 9, the last 14 amino acids of SEQ ID NO: 10, and SEQ ID NOS: 1-11. 
     
     
         83 . The composition of  claim 76 , wherein the load molecule comprises a growth factor or an anti-apoptotic agent selected from the group consisting of SEQ ID NOS: 31-34 and 46-49 inclusive, a Fibroblast growth factor (FGF), a Vascular endothelial growth factor (VEGF), Hepatocyte growth factor (HGF), a Transforming growth factor (TGF), Nerve growth factor (NGF), and a platelet derived growth factor (PDGF). 
     
     
         84 . The composition of  claim 76 , wherein the load molecule comprises a growth inhibitor selected from the group consisting of SEQ ID NOS: 50-59 and an interferon. 
     
     
         85 . The composition of  claim 76 , wherein the backbone is polylysine, and the anionic group is carboxyl. 
     
     
         86 . A method of administering a pharmaceutical composition comprising injecting a subject with a pharmaceutical composition of  claim 76 . 
     
     
         87 . A method of administering a pharmaceutical composition comprising injecting a subject with a pharmaceutical composition of  claim 80 . 
     
     
         88 . A method of treating an infection in a subject in need of such treatment comprising administering to the subject an effective amount of a composition of  claim 81 . 
     
     
         89 . A method of treating an inflammation in a subject in need of such treatment comprising administering to the subject an effective amount of a composition of  claim 82 . 
     
     
         90 . A method of  claim 89 , where the inflammation is associated with arthritis, Crohn's disease, rheumatoid arthritis, chronic inflammatory bowel disease, ulcerative colitis, multiple sclerosis, or diabetes. 
     
     
         91 . A method of treating an organ or cell damage in a subject in need of growth or apoptosis prevention comprising administering to the subject an effective amount of a composition of  claim 83 . 
     
     
         92 . A method of  claim 91 , where the organ or cell damage is caused by radiation injury, is caused by myocardial infarction, is caused by autoimmune disease, or is associated with Type I diabetes. 
     
     
         93 . A method of  claim 91 , where the organ or cell damage is in the intestine, the bone marrow, the liver, or the heart. 
     
     
         94 . A method of treating a cell-overgrowth or cancer in a subject in need of such treatment using growth inhibitor or apoptosis inducer comprising administering to the subject an effective amount of a composition according to  claim 84 . 
     
     
         95 . A method of administering a pharmaceutical composition comprising injecting a subject with a pharmaceutical composition, comprising:
 (a) a pharmaceutically acceptable excipient; and   (b) a polymer consisting of:
 (i) a polymer backbone selected from polylysine, polyornithine, polyarginine, polyglutamate, polyaspartate, polycysteine, polyserine, polythreonine, polytyrosine, chondroitin sulfate, dermatan sulfate, keratan sulfate, carageenan, pectin, fucoidan, dextran, polymethylacrylic acid, polyacrylic acid, and polyallylamine; 
 (ii) multiple protective chains covalently linked to the polymer backbone, wherein each protective chain is independently selected from a group consisting of a linear poly(ethyleneglycol) and a linear methoxy poly(ethyleneglycol), and each protective chain is singly terminally covalently linked to a monomeric unit on the polymer backbone via one covalent bond; 
 (iii) an anionic group attached to a monomeric unit on the polymer backbone by a covalent bond, wherein the anionic group is selected from the group consisting of phosphate, sulfate, sulfonate, and carboxyl, or 
 an anionic cluster comprising two or more anionic groups attached to a monomeric unit on the polymer backbone by a single covalent bond, wherein the anionic groups are each selected from the group consisting of phosphate, sulfate, sulfonate, and carboxyl; and 
   (c) a load molecule linked directly by electrostatic interaction to the anionic group or the anionic cluster of the polymer without an intermediary metal ion, wherein the load molecule:
 (i) comprises a cell penetrating peptide; 
 (ii) comprises an anionic binding domain; or 
 (iii) has an isoelectric point greater than 7.3, 
   wherein at least 86.7% of the load molecule remains linked to the anionic group or the anionic cluster in the presence of 100 mM aqueous NaCl solution.