IL-1 BETA TARGETING SPHERICAL NUCLEIC ACIDS
The invention in some aspects relates to antisense inhibitors of IL1B and related methods of use. The antisense inhibitors may be formulated as a nanoparticle or other type of three dimensional presentation format.
1 . A single-stranded modified oligonucleotide consisting of 10-30 linked nucleosides and having: a gap segment consisting of two to eight linked deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein at least some nucleosides of each wing segment comprises a modified nucleotide; wherein at least 5 of the internucleoside linkages within the gap segment and the linkages connecting the gap segment to the 3′ wing segments are phosphorothioate linkages (*); and the internucleoside linkages connecting the rest of the nucleosides of both the 5′ and 3′ wing segments are phosphodiester linkages; and wherein the nucleobase sequence of the oligonucleotide consists of 10-30 contiguous nucleobases complementary to an equal length portion of SEQ ID NO: 1, or a pharmaceutically acceptable salt thereof.
2 . The oligonucleotide of claim 1 , wherein the nucleobase sequence of the oligonucleotide is mGmUmAmGmUmGG*T*G*G*T*/iMe-dC/*mGmGmAmGmAmU (SEQ ID NO: 43).
3 . The oligonucleotide of claim 1 , wherein the nucleobase sequence of the oligonucleotide is mAmGmAmUmCmCT*C*T*T*A*G*mCmAmCmUmAmC (SEQ ID NO: 3).
4 . The oligonucleotide of claim 1 , wherein the nucleobase sequence of the oligonucleotide is mGmAmAmGmGmAG*C*A*C*T*T*mCmAmUmCmUmG (SEQ ID NO: 6).
5 . The oligonucleotide of claim 1 , wherein the nucleobase sequence of the oligonucleotide is mCmCmAmAmGmGC*C*A*C*A*G*mGmUmAmUmUmU (SEQ ID NO: 7).
6 . The oligonucleotide of claim 1 , wherein the nucleobase sequence of the oligonucleotide is mUmCmCmAmGmCT*T*G*T*T*A*mUmUmGmAmUmU (SEQ ID NO: 10).
7 . The oligonucleotide of any one of claims 1 - 6 , wherein the compound is 18 nucleotides in length.
8 . The oligonucleotide of any one of claims 1 - 7 , further comprising a molecular species at one of the ends.
9 . The oligonucleotide of claim 8 , wherein the molecular species is selected from the group consisting of a spacer, a lipid, a sterol, cholesterol, stearyl, C16 alkyl chain, bile acids, cholic acid, taurocholic acid, deoxycholate, oleyl litocholic acid, oleoyl cholenic acid, glycolipids, phospholipids, sphingolipids, isoprenoids, such as steroids, vitamins, such as vitamin E, saturated fatty acids, unsaturated fatty acids, fatty acid esters, such as triglycerides, pyrenes, porphyrines, Texaphyrine, adamantane, acridines, biotin, coumarin, fluorescein, rhodamine, Texas-Red, digoxygenin, dimethoxytrityl, t-butyldimethylsilyl, t-butyldiphenylsilyl, cyanine dyes (e.g. Cy3 or Cy5), Hoechst 33258 dye, psoralen, and ibuprofen.
10 . The oligonucleotide of claim 8 , wherein the molecular species is connected directly to the compound through a linkage selected from the group consisting of phosphodiester, phosphorothioate, methylphosphonate, and amide linkages.
11 . The oligonucleotide of claim 10 , wherein the molecular species is connected indirectly to the compound through a linker.
12 . The oligonucleotide of claim 11 , wherein the linker is a non-nucleotidic linker selected from the group consisting of abasic residues (dSpacer), oligoethyleneglycol, such as triethyleneglycol (spacer 9) or hexaethyleneglycol (spacer 18), and alkane-diol, such as butanediol.
13 . The oligonucleotide of claim 12 , wherein the 3′ end of the oligonucleotide is connected to 2 consecutive linkers that are hexaethyleneglycol (spacer 18), the first hexaethyleneglycol connected to the 3′ end of the oligonucleotide, the second hexaethyleneglycol connected to the first hexaethyleneglycol and the second hexaethyleneglycol is connected to a cholesterol.
14 . The oligonucleotide of claim 1 , wherein the gap segment consists of six linked deoxynucleosides.
15 . The oligonucleotide of claim 1 , wherein all of the nucleosides of each wing segment comprise a modified nucleotide.
16 . The oligonucleotide of claim 15 , wherein each nucleoside of each wing segment comprises a 2′O-methyl ribonucleoside (m).
17 . An oligonucleotide comprising mGmUmAmGmUmGG*T*G*G*T*/iMe-dC/*mGmGmAmGmAmU/isp18//isp18//3CholTEG/(SEQ ID NO: 43), wherein m is a 2′O methyl, and wherein * is a phosphorothioate modification.
18 . An oligonucleotide comprising mAmGmAmUmCmCT*C*T*T*A*G*mCmAmCmUmAmC/isp18//isp18//3CholTEG/(SEQ ID NO: 3), wherein m is a 2′O methyl, and wherein * is a phosphorothioate modification.
19 . An oligonucleotide comprising mGmAmAmGmGmAG*C*A*C*T*T*mCmAmUmCmUmG/isp18//isp18//3CholTEG/(SEQ ID NO: 6), wherein m is a 2′O methyl, and wherein * is a phosphorothioate modification.
20 . An oligonucleotide comprising mCmCmAmAmGmGC*C*A*C*A*G*mGmUmAmUmUmU/isp18//isp18//3CholTEG/(SEQ ID NO: 7), wherein m is a 2′O methyl, and wherein * is a phosphorothioate modification.
21 . An oligonucleotide comprising mUmCmCmAmGmCT*T*G*T*T*A*mUmUmGmAmUmU/isp18//isp18//3CholTEG/(SEQ ID NO: 10), wherein m is a 2′O methyl, and wherein * is a phosphorothioate modification.
22 . A stable self-assembling nanostructure, comprising a liposomal core comprised of a lipid bilayer and having an oligonucleotide shell comprised of an antisense oligonucleotide 18 to 21 linked nucleosides in length targeted to IL-1 beta (IL1B) and linked on the oligonucleotide 3′ end to an isp18//isp18//3CholTEG, wherein the 3CholTEG interacts with lipids in the lipid bilayer such that the oligonucleotide is radially oriented and the 5′ end of the oligonucleotide faces externally from the nanostructure.
23 . The nanostructure of claim 22 , wherein the antisense oligonucleotide is 18 nucleotides in length.
24 . The nanostructure of claim 22 or 23 , wherein IL1B has a sequence of SEQ ID NO: 1.
25 . The nanostructure of any one of claims 22 - 24 , wherein the antisense oligonucleotide has 2′O methyl modifications.
26 . The nanostructure of claim 25 , wherein the antisense oligonucleotide is selected from the group consisting of mGmUmAmGmUmGG*T*G*G*T*/iMe-dC/*mGmGmAmGmAmU (SEQ ID NO: 43), mAmGmAmUmCmCT*C*T*T*A*G*mCmAmCmUmAmC (SEQ ID NO: 3), mGmAmAmGmGmAG*C*A*C*T*T*mCmAmUmCmUmG (SEQ ID NO: 6), mCmCmAmAmGmGC*C*A*C*A*G*mGmUmAmUmUmU (SEQ ID NO: 7), and mUmCmCmAmGmCT*T*G*T*T*A*mUmUmGmAmUmU (SEQ ID NO: 10) wherein—refers to a phosphodiester bond, * refers to a phosphorothioate bond, and m refers to a 0 methyl.
27 . A method for treating a disorder, comprising:
administering to a subject having a disorder a stable self-assembling nanostructure, comprising a liposomal core comprised of a lipid bilayer and having an oligonucleotide shell comprised of an antisense oligonucleotide 18 to 21 linked nucleosides in length targeted to IL-1 beta (IL1B) and linked on the oligonucleotide 3′ end to an isp18//isp18//3CholTEG, wherein the 3CholTEG interacts with lipids in the lipid bilayer such that the oligonucleotide is radially oriented and the 5′ end of the oligonucleotide faces externally from the nanostructure. in an effective amount to treat the disorder.
28 . The method of claim 27 , wherein the disorder is an inflammatory disorder.
29 . The method of claim 27 , wherein the disorder is selected from atopic dermatitis, Epidermolysis bullosa, uveitis, Gout, Polymyalgia rheumatica, Osteoarthritis, Systemic-onset juvenile idiopathic arthritis, Schnitzler syndrome, Familial Mediterranean fever, Cryopyrin-associated periodic syndrome (CAPS), Hyper-IgD syndrome (HIDS), TNF receptor-associated periodic syndrome (TRAPS), Type 2 diabetes, Proliferative diabetic retinopathy, Wet age-related macular degeneration, Chronic obstructive pulmonary disease, Type 1 diabetes, Urticarial vasculitis, Pyoderma gangrenosum, Dry eye syndrome, Acne vulgaris, as well as improve arterial function in atherosclerosis in type 2 diabetes and reduce cardiovascular risk in type 2 diabetes.
30 . A method for reducing expression levels of IL1B in vivo, comprising:
administering to a subject a pharmaceutical composition comprising a stable self-assembling nanostructure, comprising a liposomal core comprised of a lipid bilayer and having an oligonucleotide shell comprised of an antisense oligonucleotide 18 to 21 linked nucleosides in length targeted to IL-1 beta (IL1B) and linked on the oligonucleotide 3′ end to an isp18//isp18//3CholTEG, wherein the 3CholTEG interacts with lipids in the lipid bilayer such that the oligonucleotide is radially oriented and the 5′ end of the oligonucleotide faces externally from the nanostructure in an effective amount to reduce IL1B levels in vivo.
31 . The method of claim 30 , wherein the subject is a mammal.
32 . The method of claim 30 , wherein the subject is human.
33 . A method for reducing expression levels of IL1B in vitro, comprising:
contacting a cell with a stable self-assembling nanostructure, comprising a liposomal core comprised of a lipid bilayer and having an oligonucleotide shell comprised of an antisense oligonucleotide 18 to 21 linked nucleosides in length targeted to IL-1 beta (IL1B) and linked on the oligonucleotide 3′ end to an isp18//isp18//3CholTEG, wherein the 3CholTEG interacts with lipids in the lipid bilayer such that the oligonucleotide is radially oriented and the 5′ end of the oligonucleotide faces externally from the nanostructure in an effective amount to reduce IL1B levels in vitro.
34 . The method of claim 33 , wherein the cell is a keratinocyte.