Polyconjugates for In Vivo Delivery of Polynucleotides
AssigneeArrowhead Madison Inc.
InventorsDavid B. ROZEMA, Darren H. WAKEFIELD, David L. LEWIS, Jason KLEIN, So WONG, Jon A. WOLFF, James E. HAGSTROM
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Abstract
The present invention is directed to compounds, compositions, and methods useful for delivering polynucleotides or other cell-impermeable molecules to mammalian cells. Described are polyconjugates systems that incorporate targeting, anti-opsonization, anti-aggregation, and transfection activities into small biocompatible in vivo delivery vehicles. The use of multiple reversible or labile linkages connecting component parts provides for physiologically responsive activity modulation.
Claims23
An RNA interference polynucleotide delivery polyconjugate for delivering the RNA interference polynucleotide to a mammalian cell in vivo comprising:
- a membrane inactive reversibly modified amphipathic membrane active random or block copolymer comprising the general structure represented by:
- wherein
- is a primary amine-containing monomer,
- is a hydrophobic group-containing monomer,
- are reversibly modified monomers,
- RNAi is the RNA interference polynucleotide,
- L1 is a physiologically labile linkage orthogonal to maleamate,
- M1 is —CO—C(R)═C(L)-COO− or —CO—C(L)═C(R)—COO−, wherein L is charge neutral and comprises a targeting ligand and R1 is an alkyl group,
- M2 is —CO—C(R′)═C(SS)—COO− or —CO—C(SS)═C(R′)—COO−, wherein SS is charge neutral and comprises a steric stabilizer and R′ is an alkyl group,
- x is an integer greater than or equal to zero
- n is an integer greater than zero,
- y and z are independently integers greater than or equal to zero, and the value of y+z is greater than the value of x.
A method for forming a mammalian in vivo RNA interference polynucleotide delivery polyconjugate comprising:
- a) forming a membrane active polyamine;
- b) forming a first neutral masking agent comprising a disubstituted maleic anhydride containing a targeting ligand;
- c) forming a second neutral masking agent comprising a disubstituted maleic anhydride containing a steric stabilizer;
- d) linking the RNA interference polynucleotide to the polyamine via a physiologically labile covalent bond that is orthogonal to a maleamate linkage;
- e) reversibly inhibiting membrane activity of the membrane active polyamine wherein the inhibiting consists of modifying 50% or more of the amines on the polyamine by reacting the polyamine with the first and second neutral masking agents thereby linking a plurality of targeting ligands and a plurality of steric stabilizers to the membrane active polymer via physiologically pH-labile disubstituted maleamate linkages.
Dependencies are inferred from the claim text.