NANOPARTICLES FOR DELIVERY OF NUCLEIC ACIDS AND STABLE DOUBLE-STRANDED RNA
Nanoparticles of double-stranded nucleic acid complexed about a complexing agent such as the melamine derivatives of formulae I and II, preferably forming a trimeric nucleic acid complex. In alternative embodiments, polyarginine or a polymer of Gln and Asn further complexed with the double-stranded nucleic acid complex. In a preferred embodiment, the ds nucleic acid is a double stranded RNA having 15 to 30 base pairs suitable for RNA interference. In another aspect of the invention, a ds RNA is produced in which all of the uridines are changed to 5-methyluridine. Preferably, the resultant ds RNAs have 15 to about 30 base pairs and are suitable for RNA interference.
1 . A complex comprising a double-stranded ribonucleic acid (dsRNA) and a compound capable of complexing two or more double-stranded ribonucleic acids (dsRNAs).
2 . The complex of claim 1 , further comprising a polyarginine polypeptide.
3 . The complex of claim 2 , wherein the arginine residues of the polyarginine polypeptide are a mixture of D and L isomers.
4 . The complex of claim 3 , wherein the polyarginine polypeptide contains alternating D and L isomers.
5 . The complex of claim 2 , further comprising a carbohydrate or a polypeptide domain attached to an end of the polyarginine polypeptide.
6 . The complex of claim 5 , wherein the carbohydrate is mannose or galactose.
7 . The complex of claim 5 , wherein the polypeptide domain is the TAT sequence of the human immunodeficiency virus or a portion thereof.
8 . The complex of claim 1 , further comprising a polypeptide comprising glutamine (Gln) and asparagine (Asn) residues.
9 . The complex of claim 8 , wherein the glutamine (Gln) and asparagine (Asn) residues are a mixture of D and L isomers.
10 . The complex of claim 9 , wherein the residues are alternating D and L isomers.
11 . The complex of claim 1 , further comprising a polyarginine polypeptide and a polypeptide comprising glutamine (Gln) and asparagine (Asn) residues.
12 . The complex of claim 1 , wherein the compound capable of complexing two or more dsRNAs is a melamine derivative.
13 . The complex of claim 12 , wherein the melamine derivative has the structure Formula I:
14 . The complex of claim 12 , wherein the melamine derivative has the structure Formula II:
15 . The complex of claim 1 , wherein the dsRNA is a siRNA.
16 . The complex of claim 15 , wherein the siRNA contains less than or equal to 30 nucleotide pairs.
17 . The complex of claim 15 , wherein the siRNA contains 20-25 nucleotide pairs.
18 . The complex of claim 15 , wherein the siRNA is targeted to TNF-alpha, HIV virus, Hepatitis B virus, or VEGF receptor 1.
19 . The complex of claim 15 , wherein the complex releases a siRNA intracellularly to inhibit gene expression in a cell.
20 . The complex of claim 1 , comprising particles having diameters less than 200 nanometers.
21 . The complex of claim 1 , comprising particles having diameters less than 100 nanometers.
22 . A compound made by the method of:
(a) complexing a dsRNA with a compound capable of complexing two or more dsRNAs thereby forming a first complex; and
(b) complexing the first complex with a polyarginine.
23 . The compound of claim 22 , wherein a carbohydrate or a polypeptide domain is attached to an end of the polyarginine.
24 . The compound of claim 23 , wherein the carbohydrate is mannose or galactose.
25 . The compound of claim 23 , wherein the polypeptide domain is the TAT sequence of the human immunodeficiency virus or a portion thereof.
26 . A compound made by the method of:
(a) complexing a dsRNA with a compound capable of complexing two or more dsRNAs thereby forming a first complex; and
(b) complexing the first complex with a polypeptide comprising glutamine (Gln) and asparagine (Asn) residues.
27 . The compound of claim 26 , wherein the glutamine (Gln) and asparagine (Asn) residues are a mixture of D and L isomers.
28 . The compound of claim 27 , wherein the residues are alternating D and L isomers.
29 . The compound of claim 26 , wherein the compound is further complexed with a polyarginine polypeptide.