Polynucleotides encoding ornithine transcarbamylase for the treatment of urea cycle disorders

Patent metadata

Field	Value
Publication number	US-2020299652-A1
Application number	US-201816765604-A
Country	US
Kind code	A1
Filing date	Nov 21, 2018
Priority date	Nov 22, 2017
Publication date	Sep 24, 2020
Grant date	—

Abstract

This disclosure relates mRNA therapy for the treatment of ornithine transcarbamylase deficiency (OTCD). mRNAs for use in the invention, when administered in vivo, encode human ornithine transcarbamylase (OTC), isoforms thereof, functional fragments thereof, and fusion proteins comprising OTC. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to effect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of OTC expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic ammonia associated with deficient OTC activity in subjects.

First claim

What is claimed is:

1 . A pharmaceutical composition comprising an mRNA comprising an open reading frame (ORF) encoding an ornithine transcarbamylase (OTC) polypeptide, wherein the composition when administered as a single intravenous dose to a human subject in need thereof is sufficient to:
(i) increase the level of OTC activity in liver tissue to within at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of normal OTC activity level for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (ii) increase the level of OTC activity in liver tissue at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, or at least 50-fold compared to the subject's baseline OTC activity level or a reference OTC activity level in a human subject having ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (iii) reduce RBC, plasma, serum and/or liver levels of ammonia at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% compared to the subject's baseline RBC, plasma, serum and/or liver ammonia level or a reference RBC, plasma, serum and/or liver ammonia level in a human subject having ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (iv) reduce plasma, serum, and/or urine levels of orotic acid at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% compared to the subject's baseline plasma, serum, or urine orotic acid level or a reference plasma, serum, or urine orotic level in a human subject having ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (v) reduce RBC, plasma, serum and/or liver levels of ammonia at least 1.5-fold, at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, or at least 50-fold as compared to the subject's baseline RBC, plasma, serum and/or liver ammonia level or a reference RBC, plasma, serum and/or liver ammonia level in a patient with ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (vi) reduce plasma, serum, and/or urine level of orotic acid at least 1.5-fold, at least 2-fold at least 5-fold, at least 10-fold, at least 20-fold or at least 50-fold as compared to the subject's baseline plasma, serum, and/or urine orotic acid level or a reference plasma, serum, and/or urine orotic acid level in a patient with ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration; (vii) increase body weight of the human subject by at least 2%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% of pre-treatment body weight by at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 5 days, at least 7 days, at least 14 days, at least 24 days, at least 48 days, or at least 60 days post-administration; and/or (viii) maintain body weight of the human subject to within at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of pre-treatment body weight for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 5 days, at least 7 days, at least 14 days, at least 24 days, at least 48 days, or at least 60 days post-administration.

2 . The pharmaceutical composition of claim 1 , further comprising a delivery agent.

3 . The pharmaceutical composition of claim 2 , wherein the delivery agent comprises a lipid nanoparticle comprising:
(i) Compound II, (ii) Cholesterol, and (iii) PEG-DMG or Compound I; (i) Compound VI, (ii) Cholesterol, and (iii) PEG-DMG or Compound I; (i) Compound II, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I; or (i) Compound VI, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I.

4 . The pharmaceutical composition of any one of the preceding claims, wherein the OTC polypeptide comprises the amino acid sequence set forth in SEQ ID NO:1.

5 . The pharmaceutical composition of any one of the preceding claims, wherein the ORF has at least 79%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from the group consisting of SEQ ID NOs:2 and 5-29.

6 . The pharmaceutical composition of any one of the preceding claims, wherein the mRNA comprises a microRNA (miR) binding site.

7 . The pharmaceutical composition of claim 6 , wherein the microRNA is expressed in an immune cell of hematopoietic lineage or a cell that expresses TLR7 and/or TLR8 and secretes pro-inflammatory cytokines and/or chemokines.

8 . The pharmaceutical composition of claim 6 , wherein the microRNA binding site is for a microRNA selected from the group consisting of miR-126, miR-142, miR-144, miR-146, miR-150, miR-155, miR-16, miR-21, miR-223, miR-24, miR-27, miR-26a, or any combination thereof.

9 . The pharmaceutical composition of claim 6 , wherein the microRNA binding site is for a microRNA selected from the group consisting of miR126-3p, miR-142-3p, miR-142-5p, miR-155, or any combination thereof.

10 . The pharmaceutical composition of claim 6 , wherein the microRNA binding site is a miR-142-3p binding site.

11 . The pharmaceutical composition of any one of claims 6 to 10 , wherein the microRNA binding site is located in the 3′ UTR of the mRNA.

12 . The pharmaceutical composition of any one of the preceding claims, wherein the mRNA comprises a 3′ UTR comprising a nucleic acid sequence at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to the 3′ UTR sequence set forth in SEQ ID NO:4.

13 . The pharmaceutical composition of any one of the preceding claims, wherein the mRNA comprises a 5′ UTR comprising a nucleic acid sequence at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to a 5′ UTR sequence set forth in SEQ ID NO:3.

14 . The pharmaceutical composition of any one of the preceding claims, wherein the mRNA comprises a 5′ terminal cap.

15 . The pharmaceutical composition of claim 14 , wherein the 5′ terminal cap comprises a Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′-fluoro-guanosine, 7-deaz

Assignees

• Modernatx Inc

Inventors

• Zhuo Zhijian
• Frassetto Andrea Lea
• Martini Paolo G V
• Presnyak Vladimir
• Finn Patrick

Classifications

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• C12N9/1018 Primary

  Carboxy- and carbamoyl transferases (2.1.3) · CPC title

• C12Y201/03003

  Ornithine carbamoyltransferase (2.1.3.3) · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• A61K9/5123

  Organic compounds, e.g. fats, sugars · CPC title