RNA interference in ocular indications

• US 10,184,124 B2
• Filed: 06/02/2015
• Issued: 01/22/2019
• Est. Priority Date: 03/24/2010
• Status: Active Grant

• Current Assignee: Phio Pharmaceuticals Corp.
• Original Assignee: Phio Pharmaceuticals Corp.
• Inventors: Libertine, Lyn, Khvorova, Anastasia, Salomon, William, Kamens, Joanne, Samarsky, Dmitry, Woolf, Tod M., Cardia, James, Pavco, Pamela A.
• Primary Examiner(s): Whiteman, Brian A
• Application Number:
  US14/728,653
• Publication Number:
  US 20160115482A1
• Time in Patent Office: 1,330 Days
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61K 9/0051 Ocular inserts, ocular implants
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 7/10 Antioedematous agents; Diuretics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/313 Phosphorodithioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2320/31 Combination therapy
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

The present invention relates to ocular administration of sd-rxRNA and rxRNAori molecules.

Claims

1. A method for delivering a nucleic acid to an eye of a subject in need thereof, comprising

administering to the eye of the subject an sd-rxRNA®

, in an effective amount to promote RNA interference by the sd-rxRNA®

in the eye

wherein the sd-rxRNA®

comprises a guide strand and a passenger strand, wherein the sd-rxRNA®

includes a double stranded region and a single stranded region, wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′

end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 phosphorothioate modifications, wherein at least 40% of the nucleotides of the isolated double stranded nucleic acid molecule are modified, wherein the sd-rxRNA has one end that is blunt, and wherein the sd-rxRNA is directed against a sequence comprising at least 12 contiguous nucleotides of a sequence selected from SEQ ID NOs;

1-182.

2. The method of claim 1, wherein the administration of the sd-rxRNA®

is intravitreal.

3. The method of claim 1, wherein the method is a method for treating an ocular disorder, optionally wherein the ocular disorder is selected from the group consisting of:

vascular leakage, neovascularization, age-related macular degeneration (AMD), choroidal neovascularization (wet AMD), geographic atrophy (advanced dry AMD), early-to-intermediate dry AMD, post surgical cystoid macular edema (CME), nonproliferative diabetic retinopathy (NPDR), diabetic macular edema (DME), macular edema secondary to retinal vein occlusion (RVO), proliferative diabetic retinopathy (PDR), glaucoma, neovascular glaucoma (NVG), retinopathy of prematurity (ROP), fibroproliferative retinal disease, proliferative vitreoretinopathy (PVR), epiretinal membranes/vitreomacular adhesions, retinal degenerative disease, retinitis pigmentosa, retinal vascular occlusive disorders, retinal vein occlusion, retinal artery occlusion, retinoblastoma, trabeculectomy failure due to scarring, and uveitis.

4. The method of claim 3, wherein the ocular disorder is proliferative vitreoretinopathy (PVR).

5. The method of claim 1, wherein a second sd-rxRNA®

that is directed against a gene encoding for a different protein is also administered to the eye of the subject.

6. The method of claim 5, wherein the second sd-rxRNA®

comprises an sd-rxRNA®

directed against CTGF.

7. The method of claim 1, wherein the sd-rxRNA®

is hydrophobically modified, optionally wherein the sd-rxRNA®

is linked to one or more hydrophobic conjugates and/or the sd-rxRNA®

includes at least one 5-methyl C or U modifications.

8. A method for delivering a nucleic acid to an eye of a subject in need thereof, comprising

administering to the eye of the subject an sd-rxRNA®

, in an effective amount to promote RNA interference by the sd-rxRNA®

in the eye

wherein the sd-rxRNA®

comprises a guide strand and a passenger strand, wherein the sd-rxRNA®

includes a double stranded region and a single stranded region, wherein the double stranded region is from 8-15 nucleotides long, wherein the single stranded region is at the 3′

end of the guide strand and is 4-12 nucleotides long, wherein the single stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 phosphorothioate modifications, wherein at least 40% of the nucleotides of the isolated double stranded nucleic acid molecule are modified,

wherein the sd-rxRNA is directed against a gene encoding CTGF and wherein the sense strand of the sd-rxRNA®

comprises at least 12 contiguous nucleotides of the sequence of SEQ ID NO;

967 (mU.mU. G.mC. A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl), 969 (mU.mU. G.mC. A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl), 949 (G.mC. A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA.TEG-Chl), 1011 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl), 1013 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl), 1023 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.Teg-Chl), or 1025 (G.mU. G. A.mC.mC. A. A.mA. A. G*mU*mA.Teg-Chl) and/or wherein the antisense strand of the sd-rxRNA®

comprises at least 12 contiguous nucleotides of the sequence of SEQ ID NO;

968 (P.mU.fU. A. G. A.mA. A. G. G.fU. G.fC. A. A* A*fC* A*mA*mG* G.), 970 (P.mU.fU. A. G. A.mA. A. G. G.fU. G.fC.mA. A*mA*fC* A*mA*mG* G.), 950 (P.mU.fC.fU. A. G.mA. A.mA. G. G.fU. G.mC* A*mA* A*mC* A* U.), 1012 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC* A*mC*mU*mC*mU* C.), 1014 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.fC. A.mC* A*fC*mU*fC*mU* C.), 1024 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.fC. A.mC* A*fC*mU*fC*mU* C.), or 1026 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC* A*mC*mU*mC*mU* C.).

9. The method of claim 8, wherein the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

967 (mU.mU. G.mC. A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

968 (P.mU.fU. A. G. A.mA. A. G. G.fU. G.fC. A. A* A*fC* A*mA*mG* G.), the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

969 (mU.mU. G.mC. A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

970 (P.mU.fU. A. G. A.mA. A. G. G.fU. G.fC.mA. A*mA*fC* A*mA*mG* G.), the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

949 (G.mC. A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA.TEG-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

950 (P.mU.fC.fU. A. G.mA. A.mA. G. G.fU. G.mC* A*mA* A*mC* A* U.), the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

1011 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

1012 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC* A*mC*mU*mC*mU* C.), the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

1013 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

1014 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.fC. A.mC* A*fC*mU*fC*mU* C.), the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

1023 (G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.Teg-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

1024 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.fC. A.mC* A*fC*mU*fC*mU* C.), or the sense strand of the sd-rxRNA®

comprises SEQ ID NO;

1025 (G.mU. G. A.mC.mC. A. A.mA. A. G*mU*mA.Teg-Chl) and the antisense strand of the sd-rxRNA®

comprises SEQ ID NO;

1026 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC* A*mC*mU*mC*mU* C.).

10. The method of claim 8, wherein the method is a method for treating an ocular disorder, optionally wherein the ocular disorder is selected from the group consisting of:

vascular leakage, neovascularization, age-related macular degeneration (AMD), choroidal neovascularization (wet AMD), geographic atrophy (advanced dry AMD), early-to-intermediate dry AMD, post surgical cystoid macular edema (CME), nonproliferative diabetic retinopathy (NPDR), diabetic macular edema (DME), macular edema secondary to retinal vein occlusion (RVO), proliferative diabetic retinopathy (PDR), glaucoma, neovascular glaucoma (NVG), retinopathy of prematurity (ROP), fibroproliferative retinal disease, proliferative vitreoretinopathy (PVR), epiretinal membranes/vitreomacular adhesions, retinal degenerative disease, retinitis pigmentosa, retinal vascular occlusive disorders, retinal vein occlusion, retinal artery occlusion, retinoblastoma, trabeculectomy failure due to scarring, and uveitis.

11. The method of claim 8, wherein a second sd-rxRNA®

that is directed against a gene encoding for a different protein is also administered to the eye of the subject.

12. The method of claim 11, wherein the second sd-rxRNA®

comprises an sd-rxRNA®

directed against VEGF.

13. The method of claim 12, wherein the sd-rxRNA®

is directed against a sequence selected from the sequences within Table 2, optionally wherein the sd-rxRNA®

is directed against a sequence comprising at least 12 contiguous nucleotides of a sequence selected from the sequences within Table 2.

14. The method of claim 8, wherein the sd-rxRNA®

is hydrophobically modified, optionally wherein the sd-rxRNA®

is linked to one or more hydrophobic conjugates and/or the sd-rxRNA®

includes at least one 5-methyl C or U modifications.