Multicomponent degradable cationic polymers

Abstract

Degradable polymers were synthesized that self-assemble with DNA to form particles that are effective for gene delivery. Small changes to polymer synthesis conditions, particle formulation conditions, and polymer structure provides significant changes to efficacy in a cell-type dependent manner. Polymers presented here are more effective than commercially available materials, such as LIPOFECTAMINE 2000™, FUGENE®, or polyethylenimine (PEI), for gene delivery to cancerous fibroblasts or human primary fibroblasts. The presently disclosed materials may be useful for cancer therapeutics and regenerative medicine.

That which is claimed: 
     
         1 . A compound of formula (I): 
       
         
           
           
               
               
           
         
         wherein:
 n is an integer from 1 to 10,000; 
 R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9  are each independently selected from the group consisting of hydrogen, branched and unbranched alkyl, branched and unbranched alkenyl, branched and unbranched alkynyl, aryl, halogen, hydroxyl, alkoxy, carbamoyl, carboxyl ester, carbonyldioxyl, amide, thiohydroxyl, alkylthioether, amino, alkylamino, dialkylamino, trialkylamino, cyano, ureido, a substituted alkanoyl group, cyclic, cyclic aromatic, heterocyclic, and aromatic heterocyclic groups, each of which may be substituted with at least one substituent selected from the group consisting of branched or unbranched alkyl, branched and unbranched alkenyl, branched and unbranched alkynyl, amino, alkylamino, dialkylamino, trialkylamino, aryl, ureido, heterocyclic, aromatic heterocyclic, cyclic, aromatic cyclic, halogen, hydroxyl, alkoxy, cyano, amide, carbamoyl, carboxylic acid, ester, carbonyl, carbonyldioxyl, alkylthioether, and thiohydroxyl groups; 
 wherein R 1  can be present or absent and when present the compound of formula (I) further comprises a counter ion selected from the group consisting of chloride, fluoride, bromide, iodide, sulfate, nitrate, fumarate, acetate, carbonate, stearate, laurate, and oleate; and 
 at least one of R, R′, and R″ comprise a reducible or degradable linkage, and wherein each R, R′, or R″ can independently be the same or different; 
 under the proviso that when at least one R group comprises an ester linkage of the formula —C(═O)—O— and the compound of formula (I) comprises a poly(beta-amino ester), then the compound of formula (I) must also comprise one or more of the following characteristics: 
 (a) each R group is different; 
 (b) each R″ group is different; 
 (c) each R″ group is not the same as any of R′, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 ; 
 (d) the R″ groups degrade through a different mechanism than the ester-containing R groups, wherein the degradation of the R″ group is selected from the group consisting of a bioreducible mechanism or an enzymatically degradable mechanism; and/or 
 (e) the compound of formula (I) comprises a substructure of a larger cross-linked polymer, wherein the larger cross-linked polymer comprises different properties from compound of formula (I); 
 and pharmaceutically acceptable salts thereof. 
 
       
     
     
         2 . The compound of  claim 1 , under the further proviso that if at least one R group comprises an ester linkage, then the R″ groups impart one or more of the following characteristics to the compound of formula (I): independent control of cell-specific uptake and/or intracellular delivery of a particle; independent control of endosomal buffering and endosomal escape; independent control of DNA release; triggered release of an active agent; modification of a particle surface charge; increased diffusion through a cytoplasm of a cell; increased active transport through a cytoplasm of a cell; increased nuclear import within a cell; increased transcription of an associated DNA within a cell; increased translation of an associated DNA within a cell; increased persistence of an associated therapeutic agent within a cell, wherein the therapeutic agent is selected from the group consisting of DNA, RNA, a peptide or a protein. 
     
     
         3 . The compound of  claim 1 , wherein n is an integer from 1 to 1,000. 
     
     
         4 . The compound of  claim 1 , wherein n is an integer from 1 to 100. 
     
     
         5 . The compound of  claim 1 , wherein n is an integer from 1 to 30. 
     
     
         6 . The compound of  claim 1 , wherein n is an integer from 5 to 20. 
     
     
         7 . The compound of  claim 1 , wherein n is an integer from 10 to 15. 
     
     
         8 . The compound of  claim 1 , wherein n is an integer from 1 to 10. 
     
     
         9 . The compound of  claim 1 , wherein the reducible or degradable linkage comprising R, R′, and R″ is selected from the group consisting of an ester, a disulfide, an amide, an anhydride or a linkage susceptible to enzymatic degradation, subject to the proviso of  claim 1 . 
     
     
         10 . The compound of  claim 1 , wherein R comprises a backbone of a diacrylate selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         11 . The compound of  claim 1 , wherein R′ comprises a side chain derived from compound selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         12 . The compound of  claim 1 , wherein R″ comprises an end group derived from a compound selected from the group consisting of 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         13 . The compound of  claim 1 , wherein at least one R″ group comprises a group that can independently control endosomal buffering and endosomal escape. 
     
     
         14 . The compound of  claim 13 , wherein the at least one R″ group comprises a functional group that is titratable at an endosomal pH ranging from about 5 to about 7 and/or has a pH-sensitive linkage in said pH range. 
     
     
         15 . The compound of  claim 1 , wherein at least one R″ comprises a fully synthesized primary amine-terminated oligomer. 
     
     
         16 . The compound of  claim 1 , wherein at least one R″ comprises a biomolecule selected from the group consisting of poly(ethyleneglycol) (PEG), a targeting ligand, and a labeling molecule. 
     
     
         17 . The compound of  claim 16 , wherein the targeting ligand is selected from the group consisting of a sugar, a small molecule, an antibody, an antibody fragment, and a peptide sequence. 
     
     
         18 . The compound of  claim 16 , wherein the labeling molecule is selected from the group consisting of a small molecule, a quantum dot, a nanoparticle, a fluorescent molecule, a luminescent molecule, a contrast agent. 
     
     
         19 . The compound of  claim 1 , wherein at least one R″ comprises an C 1 -C 30  alkyl chain. 
     
     
         20 . The compound of  claim 19 , wherein the alkyl chain is terminated with a functional group selected from the group consisting of —OH and —NH 2 . 
     
     
         21 . The compound of  claim 16 , wherein the PEG has a molecular weight between about 5 kDa and about 30 kDa. 
     
     
         22 . The compound of  claim 1 , wherein R″ is selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         23 . The compound of  claim 1 , wherein R′ comprises a functional group selected from the group consisting of —OH, —NH 2  and —SH. 
     
     
         24 . The compound of  claim 1 , wherein the compound of formula (I) has a backbone comprising ester linkages and has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         25 . The compound of  claim 1 , wherein the compound of formula (I) has a backbone comprising disulfide linkages and has the following structure: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  each independently are C 1 -C 30  alkyl chains and R″ comprises a non-reducible amino group independent from the structure of R′ or —C—R′. 
       
     
     
         26 . The compound of  claim 25 , wherein the non-reducible R″ group is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         27 . The compound of  claim 26 , wherein R′ comprises a hydroxyl group. 
     
     
         28 . A copolymer of a compound of formula (I), wherein the copolymer has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         29 . The copolymer of  claim 28 , wherein the copolymer comprises one or more monomers selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         30 . A copolymer/branched network polymer comprising a compound of  claim 1  and wherein the copolymer further comprises at least one free amine group. 
     
     
         31 . The compound of  claim 1 , wherein the compound of formula (I) has a structure selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         32 . A pharmaceutical composition comprising a compound of any of  claims 1 - 31 . 
     
     
         33 . The pharmaceutical composition of  claim 32  further comprising a therapeutic agent. 
     
     
         34 . The pharmaceutical composition of  claim 33 , wherein the therapeutic agent is selected from the group consisting of a gene, DNA, RNA, siRNA, miRNA, isRNA, agRNA, smRNA, a nucleic acid, a peptide, a protein, a chemotherapeutic agent, a hydrophobic drug, and a small molecule drug. 
     
     
         35 . The pharmaceutical composition of any of  claims 32 - 34  further comprising a nanoparticle or microparticle comprising the compound of any of  claims 1 - 31 . 
     
     
         36 . A method of treating a disease or condition, the method comprising administering to a subject in need of treatment thereof, a compound of any of  claims 1 - 31  or a pharmaceutical composition of  claims 32 - 35  comprising a therapeutic agent effective for treating the disease or condition. 
     
     
         37 . The method of  claim 36 , where in the disease or condition is selected from the group consisting of cancer, including brain cancer (including Glioblastoma Multiforme), lung cancer, breast cancer, prostate cancer, colorectal cancer, and other cancers; cardiovascular diseases; infectious diseases; ophthalmic diseases, including age-related macular degeneration. 
     
     
         38 . A method of delivering a therapeutic agent to a cell, a specific cell line, a tissue, or an organism, the method comprising associating the therapeutic agent with a compound of any of  claims 1 - 31  or a pharmaceutical composition of  claims 32 - 35  to form one or more particles comprising the agent and compound of  claims 1 - 31 , and administering the one or more particles or contacting the one or more particles with the cell, specific cell line, tissue or organism. 
     
     
         39 . The method of  claim 38 , wherein the therapeutic agent is selected from the group consisting of a gene, DNA, RNA, siRNA, miRNA, isRNA, agRNA, smRNA, a nucleic acid, a peptide, a protein, a chemotherapeutic agent, a hydrophobic drug, and a small molecule drug. 
     
     
         40 . An in vitro kit comprising a compound of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35 . 
     
     
         41 . A biomedical device comprising a compound of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35 . 
     
     
         42 . The biomedical device of  claim 41 , wherein the device comprises a stent or a stent-like device. 
     
     
         43 . An article coated with one or more compounds of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35 . 
     
     
         44 . The article of  claim 43 , wherein the article is coated with a single layer of one or more compounds of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35 . 
     
     
         45 . The article of  claim 43 , wherein the article is coated with more than one layer of one or more compounds of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35 . 
     
     
         46 . The article of  claim 45 , wherein the more than one layer comprises a layer-by-layer polyelectrolyte coat. 
     
     
         47 . The article of any of  claims 43 - 46 , further comprising coating the article with a combination of a compound of any of  claims 1 - 31  or a pharmaceutical composition of any of  claims 32 - 35  and one or more commercially available and/or FDA-approved polyelectrolyte. 
     
     
         48 . The article of any of  claims 43 - 47 , wherein the article is selected from the group consisting of a nanoparticle, a microparticle, a stent, and a stent-like devices. 
     
     
         49 . A method of forming a tissue scaffolding structure, the method comprising implanting into a subject a polymeric matrix comprising a compound of any one of  claims 1 - 31 . 
     
     
         50 . The method of  claim 49 , wherein the polymeric matrix has a porosity and a median pore size sufficient to allow for vascular ingrowth and the introduction of cells into the matrix without damage to the cells or subject. 
     
     
         51 . The method of  claim 49 , further comprising introducing one or more cells into the polymeric matrix after the matrix is implanted in the subject. 
     
     
         52 . The method of  claim 51 , wherein the cells are selected from the group consisting of hepatocytes, pancreatic islet cells, fibroblasts, chondrocytes, osteoblasts, exocrine cells, cells of intestinal origin, bile duct cells, parathyroid cells, thyroid cells, cells of the adrenal-hypothalamic-pituitary axis, heart muscle cells, epithelial cells, kidney tubular cells, kidney basement cells, kidney tubular cells, kidney basement membrane cells, nerve cells, blood vessel cells, cells forming bone and cartilage, smooth and skeletal muscle cells, cells from the retina and other parts of the eye, stem cells, induced pluripotent stem cells, and three-dimensional organoids. 
     
     
         53 . A nanoparticle or microparticle comprising a compound of any of  claims 1 - 31 . 
     
     
         54 . The nanoparticle or microparticle of  claim 53 , wherein the nanoparticle or microparticle has at least one dimension ranging from about 1 nm to about 300 nm. 
     
     
         55 . The nanoparticle or microparticle of  claim 54 , wherein the nanoparticle or microparticle has at least one dimension of about 100 nm. 
     
     
         56 . A method of using and storing the compounds of any of  claims 1 - 31  or the compositions of any of  claims 32 - 35  or the particles of any of  claims 53 - 55 , the method comprising adding a cryoprotectant to the compound, composition, and/or particles to form a mixture and lyophilizing the mixture to form storable powder of the compound, composition and/or particles. 
     
     
         57 . The method of  claim 56 , wherein the cyroprotectant comprises a sugar.