Compositions and methods for polynucleotide delivery

A method of condensing a polynucleotide, said method comprising:

• contacting the polynucleotide with an amount of a polycationic agent sufficient to reduce the size of said polynucleotide;
• said polycationic agent having the following formula:
• wherein
• n is an integer from 10 to 100 resulting in a repeating trimer wherein two R1 groups of each trimer are neutral and one R1 group of each trimer is cationic;
• R2 and R3 are hydrogen for each monomer:
• Ta and Tc are terminating groups;
• said cationic R1 groups are selected from the group consisting of ammoniumalkyl, aminoalkyl, guanidinoalkyl, amidinoalkyl, aminocyclohexyl, piperidyl, guanidinobenzyl, guanidino-α-methylbenzyl, amidinobenzyl, amidino-α-methylbenzyl, an aliphatic hydrazide, an aromatic hydrazide;
• pyridylmethyl, wherein the pyridyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• and aminobenzyl, wherein the benzyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• wherein “ammonium” is trimethylammonium, triethylammonium, methyldiethylammonium, or ethyldimethylammonium, and “alkyl” is —CH(R) -(CH2)m-, where R is H or Me, and m is 1-6;
• and said neutral R1 groups are selected from the group consisting of a neutral moiety comprising an aromatic ring and having a molecular weight of 250 daltons or less, cyclohexyl methyl, 2-methoxyethyl, and 6-galactosyl;
• wherein said polycationic agent comprises at least 25% positively charged monomers, excluding the terminal groups, and
• wherein said polycationic agent exhibits a net positive electrical charge at a physiological pH, and is capable of mediating entry of polynucleotides into a cell.

A method of inhibiting serum degradation of a polynucleotide, said method comprising contacting the polynucleotide with an amount of a polycationic agent effective to inhibit serum degradation for at least 10 minutes as compared to unprotected polynucleotide,

• said polycationic agent having the following formula:
• wherein
• n is an integer from 10 to 100 resulting in a repeating trimer wherein two R1 groups of each trimer are neutral and one R1 group of each trimer is cationic;
• R2 and R3 are hydrogen for each monomer:
• Ta and Tc are terminating groups;
• said cationic R1 groups are selected from the group consisting of ammoniumalkyl, aminoalkyl, guanidinoalkyl, amidinoalkyl, aminocyclohexyl, piperidyl, guanidinobenzyl, guanidino-α-methylbenzyl, amidinobenzyl, amidino-α-methylbenzyl, an aliphatic hydrazide, an aromatic hydrazide;
• pyridylmethyl, wherein the pyridyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• and aminobenzyl, wherein the benzyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• wherein “ammonium” is trimethylammonium, triethylammonium, methyldiethylammonium, or ethyldimethylammonium, and “alkyl” is —CH(R) -(CH2)m-, where R is H or Me, and m is 1-6;
• and said neutral R1 groups are selected from the group consisting of a neutral moiety comprising an aromatic ring and having a molecular weight of 250 daltons or less, cyclohexyl methyl, 2-methoxyethyl, and 6-galactosyl;
• wherein said polycationic agent comprises at least 25% positively charged monomers, excluding the terminal groups, and
• wherein said polycationic agent exhibits a net positive electrical charge at a physiological pH, and is capable of mediating entry of polynucleotides into a cell.

A method of producing a composition for facilitating entry of a polynucleotide into a cell, said method comprising:

• (i) contacting a polynucleotide with a polycationic agent, in an amount effective to neutralize said polynucleotide, to form a complex;
• and
• (ii) contacting the complex with a lipoprotein;
• said polycationic agent having the following formula:
• wherein
• n is an integer from 10 to 100 resulting in a repeating trimer wherein two R1 groups of each trimer are neutral and one R1 group of each trimer is cationic;
• R2 and R3 are hydrogen for each monomer:
• Ta and Tc are terminating groups;
• said cationic R1 groups are selected from the group consisting of ammoniumalkyl, aminoalkyl, guanidinoalkyl, amidinoalkyl, aminocyclohexyl, piperidyl, guanidinobenzyl, guanidino-α-methylbenzyl, amidinobenzyl, amidino-α-methylbenzyl, an aliphatic hydrazide, an aromatic hydrazide;
• pyridylmethyl, wherein the pyridyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• and aminobenzyl, wherein the benzyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• wherein “ammonium” is trimethylammonium, triethylammonium, methyldiethylammonium, or ethyldimethylammonium, and “alkyl” is —CH(R) -(CH2)m-, where R is H or Me, and m is 1-6;
• and said neutral R1 groups are selected from the group consisting of a neutral moiety comprising an aromatic ring and having a molecular weight of 250 daltons or less, cyclohexyl methyl, 2-methoxyethyl, and 6-galactosyl;
• wherein said polycationic agent comprises at least 25% positively charged monomers, excluding the terminal groups, and
• wherein said polycationic agent exhibits a net positive electrical charge at a physiological pH, and is capable of mediating entry of polynucleotides into a cell.

A method of increasing the frequency of polynucleotide uptake into a cell, said method comprising:

• (i) providing a composition that comprises
• (a) a polynucleotide;
• (b) a polycationic agent in an amount effective to neutralize said polynucleotide;
• and
• (c) an amount of lipoprotein effective to increase said frequency;
• and
• (ii) contacting said composition with said cell;
• said polycationic agent having the following formula:
• wherein
• n is an integer from 10 to 100 resulting in a repeating trimer wherein two R1 groups of each trimer are neutral and one R1 group of each trimer is cationic;
• R2 and R3 are hydrogen for each monomer:
• Ta and Tc are terminating groups;
• said cationic R1 groups are selected from the group consisting of ammoniumalkyl, aminoalkyl, guanidinoalkyl, amidinoalkyl, aminocyclohexyl, piperidyl, guanidinobenzyl, guanidino-α-methylbenzyl, amidinobenzyl, amidino-α-methylbenzyl, an aliphatic hydrazide, an aromatic hydrazide;
• pyridylmethyl, wherein the pyridyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• and aminobenzyl, wherein the benzyl ring may be further substituted with methyl, methoxy, fluoro, chloro, bromo, or nitro;
• wherein “ammonium” is trimethylammonium, triethylammonium, methyldiethylammonium, or ethyldimethylammonium, and “alkyl” is —CH(R) -(CH2)m-, where R is H or Me, and m is 1-6;
• and said neutral R1 groups are selected from the group consisting of a neutral moiety comprising an aromatic ring and having a molecular weight of 250 daltons or less, cyclohexyl methyl, 2-methoxyethyl, and 6-galactosyl;
• wherein said polycationic agent comprises at least 25% positively charged monomers, excluding the terminal groups, and
• wherein said polycationic agent exhibits a net positive electrical charge at a physiological pH, and is capable of mediating entry of polynucleotides into a cell.