Bioactive conjugates for oligonucleotide delivery
Provided herein are self-delivering oligonucleotides that are characterized by efficient RISC entry, minimum immune response and off-target effects, efficient cellular uptake without formulation, and efficient and specific tissue distribution.
1. A method for selectively delivering a compound to the placenta of a patient, comprising administering said compound to the patient, wherein the compound has a formula (I):
wherein:
O is a double-stranded nucleic acid comprising a first oligonucleotide and a second oligonucleotide, wherein:
(1) the first oligonucleotide comprises at least 16 contiguous nucleotides, a 5′ end, a 3′ end and has complementarity to a target;
(2) the second oligonucleotide comprises at least 15 contiguous nucleotides, a 5′ end, a 3′ end, and has homology with a target; and
(3) a portion of the first oligonucleotide is complementary to a portion of the second oligonucleotide;
L is a trivalent linker;
X c is a hydrophobic moiety; and
Z c is a phosphodiester or phosphodiester derivative, or is absent,
wherein X c comprises docosanoic acid (DCA) and Z c is
wherein X is O, S or BH 3 .
2. A method for treating a disease or disorder of the placenta in a patient in need of such treatment, comprising administering to the patient a small interfering RNA (siRNA) of formula (I):
wherein:
O is a double-stranded nucleic acid comprising a first oligonucleotide and a second oligonucleotide, wherein:
(1) the first oligonucleotide comprises at least 16 contiguous nucleotides, a 5′ end, a 3′ end and has complementarity to a target;
(2) the second oligonucleotide comprises at least 15 contiguous nucleotides, a 5′ end, a 3′ end, and has homology with a target; and
(3) a portion of the first oligonucleotide is complementary to a portion of the second oligonucleotide;
L is a trivalent linker;
X c is a hydrophobic moiety; and
Z c is
wherein X is O, S or BH 3 ,
wherein X c comprises docosanoic acid (DCA), and
wherein the disease or disorder is selected from the group consisting of: a disorder associated with the expression of soluble Flt1 (sFlt1) protein, PE (preeclampsia), postpartum PE, eclampsia, and HELLP syndrome.
3. The method of claim 1 , wherein L comprises an ethylene glycol chain, an alkyl chain, a peptide, RNA, DNA, a phosphodiester, a phosphorothioate, a phosphoramidate, an amide, a carbamate, or a combination thereof; and wherein L is attached to O via the second oligonucleotide.
4. The method of claim 1 , wherein O comprises one or more chemically-modified nucleotides.
5. The method of claim 1 , wherein the nucleotides at positions 2 and 14 from the 5′ end of the second oligonucleotide are 2′-methoxy-ribonucleotides.
6. The method of claim 1 , wherein:
the nucleotides at positions 1 and 2 from the 3′ end of the second oligonucleotide are connected to adjacent nucleotides via phosphorothioate linkages; and/or
the nucleotides at positions 1 and 2 from the 5′ end of the second oligonucleotide are connected to adjacent nucleotides via phosphorothioate linkages.
7. The method of claim 1 , wherein the first oligonucleotide comprises a moiety X at the 5′ end, wherein X is selected from the group consisting of
8. The method of claim 7 , wherein:
X is X3; and
L is
9. The method of claim 1 , wherein the first oligonucleotide has 3-7 more nucleotides than the second oligonucleotide.
10. The method of claim 1 , wherein:
the first oligonucleotide has perfect complementarity to the target;
the second oligonucleotide has complete homology with the target; and/or
the target is mammalian or viral mRNA, wherein the target is optionally an intronic region of said mRNA.
11. The method of claim 2 , wherein the siRNA targets a soluble fms-like tyrosine kinase-1 (sFlt-1) mRNA.