Hybrid carriers for nucleic acid cargo

Abstract

A composition for the delivery of a nucleic acid compound is provided which comprises a cationic peptide or polymer and a lipidoid compound. The nucleic acid compound may be any chemically modified or unmodified DNA or RNA. The amount of the lipidoid in the composition is preferably low, relative to the cationic peptide or polymer.

1 . A composition comprising:
 (a) a cationic peptide or polymer;   (b) a cationic or permanent lipidoid compound; and   (c) a nucleic acid compound;   
       wherein the cationic or permanent cationic lipidoid compound comprises two or three moieties of formula IIa and/or formula IIb:
   —N(R 1 )—CH 2 —CH(R 5 )—R 2   (formula IIa)
 
   —N + (R 3 )(R 4 )—CH 2 —CH(R 5 )—R 2   (formula IIb)
 
 
       wherein independently for each individual moiety of formula IIa or formula IIb
 R 1  is selected from hydrogen or C 1 -C 4 -alkyl, 
 R 2  is selected from linear or branched, saturated or unsaturated C 6 -C 16  hydrocarbyl chain, 
 R 3  and R 4  are selected from C 1 -C 4 -alkyl, and 
 R 5  is hydrogen or hydroxyl. 
 
     
     
         2 - 8 . (canceled) 
     
     
         9 . The composition of  claim 1 , wherein the cationic peptide or polymer is a compound comprising at least one cationic moiety P having at least one —SH group capable of forming a disulfide linkage, or a disulfide-linked multimer thereof, wherein moiety P is selected from
 a polymer moiety having a molecular weight from about 0.5 kDa to about 30 kDa, or 
 a peptide moiety composed of 3 to 100 amino acids, wherein at least 10% of the total number of amino acids of the peptide moiety represent basic amino acids selected from Arg, Lys, His and/or Orn. 
 
     
     
         10 . The composition of  claim 9 , wherein moiety P is
 a peptide moiety composed of 7 to 30 amino acids, and wherein the at least one —SH group is provided by a Cys residue; or   a polymer moiety selected from an optionally modified polyacrylate, chitosan, polyethylenimine, polyamine, polyaminoesters, or polyamidoamine, or any copolymer thereof.   
     
     
         11 . The composition of  claim 10 , wherein the peptide moiety has two terminal ends, and wherein—the Cys residue is located at, or in proximity to, one of the terminal ends; or wherein—the peptide moiety comprises at least two Cys residues, and wherein at least one of the Cys residues is located at, or in proximity to, each of the terminal ends. 
     
     
         12 . The composition of  claim 9 , wherein the cationic peptide or polymer is a compound according to formula
   L 1 -P 1 -[P-] n -P 3 -L 2   (formula IV)
   
       wherein
 P is defined as above; 
 P 3  is optional; 
 P 1  and P 3  are independently selected, each representing a linear or branched hydrophilic polymer chain selected from polyethylene glycol (PEG), poly-N-(2-hydroxypropyl)methacrylamide, poly-2-(methacryloyloxy)ethyl phosphorylcholines, poly(hydroxyalkyl L-asparagine), poly(2-(methacryloyloxy)ethyl phosphorylcholine), hydroxyethylstarch or poly(hydroxyalkyl L-glutamine), wherein the polymer chain exhibits a molecular weight from about 1 kDa to about 100 kDa, and wherein each of P 1  and P 3  is linked with a moiety P through a disulfide linkage; 
 L 1  and L 2  are optional ligands and independently selected from RGD, an RGD peptide, transferrin, folate, a signal peptide or signal sequence, a localization signal or sequence, a nuclear localization signal or sequence (NLS), an antibody, a cell penetrating peptide such as WEAKLAKALAKALAKHLAKALAKALKACEA (SEQ ID NO:764), TAT, a ligand of a receptor, cytokine, hormone, growth factor, small molecule, carbohydrate, mannose, galactose, n-acetylgalactosamine, synthetic ligand, small molecule agonist, inhibitor or antagonist of a receptor, or a RGD peptidomimetic analogue; 
 n is an integer, selected from 1 to about 50, preferably in the range from 2, 3, 4, or 5 to about 10, or from 2, 3, or 4 to about 9, such as 6, or 7; 
 and wherein, if n is greater than 1, each moiety P is linked with another moiety P through a disulfide linkage. 
 
     
     
         13 . The composition of  claim 1 , wherein the weight ratio of the cationic peptide or polymer to the nucleic acid compound is at least about 1, and wherein the ratio of the lipidoid compound to the nucleic acid compound is not higher than about 15 nmol/μg. 
     
     
         14 . The composition of  claim 1 , wherein the weight ratio of the lipidoid compound to the cationic peptide or polymer is not higher than about 1:50, and/or wherein the ratio of the lipidoid compound to the cationic peptide or polymer is not higher than about 2 nmol/μg. 
     
     
         15 . The composition of  claim 1 , having an N/P ratio from about 0.1 to about 20, or from about 0.2 to about 15, or from about 2 to about 15, or from about 2 to about 12, wherein the N/P ratio is defined as the mole ratio of the nitrogen atoms of the basic groups of the cationic peptide or polymer to the phosphate groups of the nucleic acid compound. 
     
     
         16 . (canceled) 
     
     
         17 . The composition of  claim 1 , comprising two or more different species of cationic peptides and/or polymers. 
     
     
         18 . The composition of  claim 1 , wherein the nucleic acid compound is selected from
 chemically modified or unmodified DNA, single stranded or double stranded DNA, coding or non-coding DNA, optionally selected from plasmid, oligodesoxynucleotide, genomic DNA, DNA primers, DNA probes, immunostimulatory DNA, aptamer, or any combination thereof, and/or   chemically modified or unmodified RNA, single-stranded or double-stranded RNA, coding or non-coding RNA, optionally selected from messenger RNA (mRNA), oligoribonucleotide, viral RNA, replicon RNA, transfer RNA (tRNA), ribosomal RNA (rRNA), immunostimulatory RNA (isRNA), microRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small-hairpin RNA (shRNA) or a riboswitch, an RNA aptamer, an RNA decoy, an antisense RNA, a ribozyme, or any combination thereof.   
     
     
         19 . The composition of  claim 1 , further comprising one or more compounds independently selected from targeting agents, cell penetrating agents, and stealth agents. 
     
     
         20 . The composition of  claim 1 , wherein the cationic permanent cationic lipidoid compound is 3-C12, 3-C12-OH, or 3-C12-OH-cat, optionally further comprising a pharmaceutically acceptable anion. 
     
     
         21 . A nanoparticle comprising a composition of  claim 1 . 
     
     
         22 - 31 . (canceled) 
     
     
         32 . A method of prophylaxis, treatment, and/or amelioration of a disease or disorder in a patient in need thereof comprising administering to the patient an effective amount of the composition of  claim 1 . 
     
     
         33 . The method of  claim 32 , wherein the disease is selected from cancer or tumour diseases, infectious diseases, autoimmune diseases, allergies or allergic diseases, monogenetic diseases, i.e. (hereditary) diseases, or genetic diseases in general, diseases which have a genetic inherited background and which are typically caused by a defined gene defect and are inherited according to Mendel's laws, cardiovascular diseases, neuronal diseases, diseases of the respiratory system, diseases of the digestive system, diseases of the skin, musculoskeletal disorders, disorders of the connective tissue, neoplasms, immune deficiencies, endocrine, nutritional and metabolic diseases, eye diseases, ear diseases and diseases associated with a peptide or protein deficiency. 
     
     
         34 - 38 . (canceled) 
     
     
         39 . The method of  claim 32 , wherein the composition is administered via ocular delivery. 
     
     
         40 - 42 . (canceled) 
     
     
         43 . The method of  claim 39 , wherein the ocular delivery is intravitreal, intracameral, subconjunctival, subretinal, subtenon, retrobulbar, topical, and/or posterior juxtascleral administration or the ciliary muscle. 
     
     
         44 . The method of  claim 33 , wherein the disease is a viral, bacterial, or protozoological infectious disease. 
     
     
         45 . The method of  claim 43 , wherein the ocular delivery is into the ciliary muscle. 
     
     
         46 . The method of  claim 43 , wherein the nanoparticle or the composition comprises an mRNA encoding a protein such that the administration of the nanoparticle or the composition results in expression and/or activity of the protein encoded by the mRNA in the eye.