RNAi CONJUGATES, PARTICLES AND FORMULATIONS THEREOF
Particles, including nanoparticles and microparticles, and pharmaceutical formulations thereof, comprising conjugates of an RNAi agent attached to a targeting moiety via a linker have been designed which can provide improved temporospatial delivery of the RNAi agent and/or improved biodistribution. Methods of making the conjugates, the particles, and the formulations thereof are provided. Methods of administering the formulations to a subject in need thereof are provided, for example, to treat or prevent cancer or infectious diseases.
1 . A conjugate comprising an RNA interference (RNAi) agent coupled to a targeting moiety by a linker.
2 . The conjugate of claim 1 , wherein the conjugate comprises a formula selected from the group X—Y—Z, X—Y—Z—Y—X, X—Y—Zn, and (X—Y—Z—Y) n —Z;
wherein X is the targeting moiety,
Y is the linker,
Z is the RNAi agent, and
n is an integer between 2 and 1,000.
3 . The conjugate of claim 1 , wherein the conjugate comprises the formula X—Y—Z;
wherein X is the targeting moiety,
Y is the linker, and
Z is the RNAi agent.
4 . The conjugate of claims 1 , wherein the conjugate comprises Formula 1a:
wherein X is the targeting moiety; Z is the RNAi agent; X′ is either absent or independently selected from carbonyl, amide, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, one or more natural or unnatural amino acids, thio or succinimido; R 1 and R 2 are either absent or comprised of alkyl, substituted alkyl, aryl, substituted aryl, polyethylene glycol (2-30 units); Y′ is absent, substituted or unsubstituted 1,2-diaminoethane, polyethylene glycol (2-30 units) or an amide; Z′ is either absent or independently selected from carbonyl, amide, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, thio or succinimido.
5 . The conjugate of claim 1 , wherein the conjugate comprises Formula Ib:
wherein X is the targeting moiety, Z is the RNAi agent, m=0-20, and A is a spacer unit, either absent or independently selected from the following substituents:
wherein z=0-40, R is H or an optionally substituted alkyl group, and R′ is any side chain found in either natural or unnatural amino acids.
6 . The conjugate of claim 1 , wherein the conjugate comprises Formula Ic:
wherein X is the targeting moiety, Z is the RNAi agent, C is a branched unit containing three to six functionalities selected from amines, carboxylic acids, thiols, or succinimides, including amino acids such as lysine, 2,3-diaminopropanoic acid, 2,4-diaminobutyric acid, glutamic acid, aspartic acid, and cysteine, m=0-40, n=0-40, x=1-5, y=1-5, and A is a spacer unit, either absent or independently selected from the following substituents:
wherein z=0-40, R is H or an optionally substituted alkyl group, and R′ is any side chain found in either natural or unnatural amino acids.
7 . The conjugate of claim 1 , wherein the linker is not a cleavable linker.
8 . The conjugate of claim 1 , wherein the linker is a cleavable linker.
9 . The conjugate of claim 8 , wherein the linker is cleavable in cytoplasm, endosome or lysosome.
10 . The conjugate of claim 1 , wherein the linker comprises an ester bond, disulfide, amide, acylhydrazone, ether, carbamate, carbonate, or urea.
11 . The conjugate of claim 1 , wherein the linker comprises a cell-penetrating peptide.
12 . The conjugate of claim 1 , wherein the RNAi agent is selected from small-interfering RNA (siRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small-activating RNA (saRNA) or combinations thereof.
13 . The conjugate of claim 1 , wherein the RNAi agent is a double-stranded RNA (dsRNA).
14 . The conjugate of claim 1 , wherein the RNAi agent is a single-stranded RNA (ssRNA).
15 . The conjugate of claim 1 , wherein the targeting moiety is selected from peptides, antibody mimetics, aptamers, antibodies, glycoproteins, small molecules, carbohydrates, or lipids.
16 . The conjugate of claims 15 , wherein the targeting moiety binds to a cell-surface receptor.
17 . The conjugate of claim 15 , wherein the targeting moiety binds to cancer cells.
18 . The conjugate of claim 15 , wherein the targeting moiety binds to viral cells.
19 .- 21 . (canceled)
22 . A polymeric particle comprising the conjugate of claim 1 and at least one polymeric matrix.
23 . The particle of claim 22 , wherein the polymeric matrix comprises one or more polymers selected from the group consisting of hydrophobic polymers, hydrophilic polymers, and copolymers thereof.
24 .- 25 . (canceled)
26 . The particle of claim 22 , wherein the polymeric matrix comprises one or more polymers selected from the group consisting of poly(lactic acid), poly(glycolic acid), poly(lactic-co-glycolic acid), poly(ethylene oxide), poly(ethylene glycol), poly(propylene glycol), and copolymers thereof.
27 . The particle of claim 22 , wherein the particle has a diameter between 10 nm and 5000 nm.
28 .- 33 . (canceled)
34 . The particle of claim 22 , wherein the conjugate is present in an amount between 0.05% and 50% (w/w) based upon the weight of the particle.
35 . The conjugate of claim 1 , wherein the conjugate has a molecular weight of less than 50,000 Da.
36 . The conjugate of claim 35 , wherein the conjugate has a molecular weight of between about 1000 Da and about 5000 Da.
37 . A pharmaceutical formulation comprising the conjugate of claim 1 and at least one pharmaceutically acceptable excipient.
38 . A method of treating a subject in need thereof comprising administering a therapeutically effective amount of the formulation of claim 37 .
39 . The method of claim 38 , wherein the subject has cancer.
40 . The method of claim 38 , wherein the subject has viral infection.
41 .- 45 . (canceled)