# US 2010/0168213 A1 · Protein-coding RNA to correct mitochondrial dysfunction

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- Number: US 2010/0168213 A1
- Document ID: US20100168213A1
- Status: Published application
- Kind: A1
- Application number: 12/584,059
- Filed: 2009-08-28
- Published: 2010-07-01
- Earliest claimed: 2008-08-28
- Assignees: Council of Scientific and Industrial Research
- Inventors: Samit Adhya
- Web page: https://patentgrep.com/patent/US20100168213A1

## CPC

- C07K 14/4707: {Muscular dystrophy}
- A61K 48/0008: {characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition}
- A61K 48/0025: {wherein the non-active part clearly interacts with the delivered nucleic acid}
- A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered}
- C07K 2319/07: containing a mitochondrial localisation signal
- C12N 2810/10: Vectors comprising a non-peptidic targeting moiety
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides compositions and methods for using correctional RNA, with a proteinaceous carrier, to stimulate respiration of the cells, tissues, organs or the whole organism of normal or diseased subjects. In one embodiment of the invention, the signal-tagged correctional RNA is one or more protein-coding RNAs (pcRNA) that encode one or more of human mitochondrial proteins. The invention specifically provides a broad-spectrum formulation applicable to a wide variety of disorders that are associated with mitochondrial mutations.

## Claims

### Claim 1

A recombinant polynucleotide segment comprising a sequence set forth in SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11, wherein the sequence comprises an operably linked signal tag.

### Claim 2 (depends on 1)

The recombinant polynucleotide segment of claim 1, wherein the sequence is the one set forth in SEQ ID NO: 9.

### Claim 3 (depends on 1)

The recombinant polynucleotide segment of claim 1, wherein the sequence is the one set forth in SEQ ID NO: 10.

### Claim 4 (depends on 1)

The recombinant polynucleotide segment of claim 1, wherein the sequence is the one set forth in SEQ ID NO: 11.

### Claim 5 (depends on 1)

A vector comprising the recombinant polynucleotide segment of claim 1, wherein the vector comprises: a non-nuclear nucleic acid sequence encoding the peptide.

### Claim 6 (depends on 5)

An isolated host cell transformed with the vector of claim 5, wherein the cell is of animal, plant or protest origin.

### Claim 7 (depends on 4)

A process for producing at least one human mitochondrial protein intra-cellularly, the process comprising the steps of culturing the isolated host cell of claim 4 in the presence of a group of proteins comprising SEQ ID NOS: 1, 3, 5, 6, 7 and 8 under conditions sufficient for the production of at least one human mitochondrial protein.

### Claim 8 (depends on 7)

The process of claim 7, wherein the group of proteins is SEQ ID NOS: 1, 3, 5, 6, 7 and 8.

### Claim 9 (depends on 7)

The process claim 7, wherein the group of proteins is SEQ ID NOS: 1-8.

### Claim 10

A recombinant polynucleotide comprising a sequence set forth in SEQ ID NO:12 and a complementary strand thereof selected from the group consisting of SEQ ID NO:13 and SEQ ID NO:14.

### Claim 11

A composition comprising a polyribonucleotide sequence set forth in SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11 or a mixture thereof.

### Claim 12 (depends on 11)

The composition of claim 11, wherein the composition comprises polyribonucleotide sequences set forth in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11.

### Claim 13 (depends on 11)

The composition of claim 11 further comprising a carrier that is a protein or a group of proteins for transporting the polyribonucleotide sequence or the mixture thereof across cellular and mitochondrial membranes.

### Claim 14 (depends on 11)

The composition of claim 11, wherein the group of proteins comprises SEQ ID NOS: 1, 3, 5, 6, 7 and 8.

### Claim 15 (depends on 11)

The composition of claim 11, wherein the group of proteins comprises SEQ ID NOS: 1-8.

### Claim 16 (depends on 11)

The composition of claim 11, further comprising a binding buffer.

### Claim 17

Use of composition comprising an amount of a polyribonucleotide sequence set forth in SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11 or a mixture thereof, a carrier comprising a group of proteins comprising SEQ ID NOS: 1, 3, 5, 6, 7 and 8 and a binding buffer in the preparation of a medicament for the treatment of the disease or physiological condition associated with mitochondrial dysfunction.

### Claim 18

A method of treating a disease or physiological condition associated with mitochondrial dysfunction in a mammal comprising administering a composition comprising an amount of a polyribonucleotide sequence set forth in SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11 or a mixture thereof, a carrier comprising a group of proteins comprising SEQ ID NOS: 1, 3, 5, 6, 7 and 8 and a binding buffer sufficient for the treatment of the disease or physiological condition associated with mitochondrial dysfunction.

### Claim 19 (depends on 18)

The method of claim 18, wherein the carrier is administered separately together with the polyribonucleotide sequence or mixture thereof in a pharmaceutically acceptable formulation.

### Claim 20 (depends on 19)

The method of claim 19, wherein the formulation is selected from the group comprising of an aqueous solution and water- or oil-based ointments, creams or suppositories.

### Claim 21 (depends on 18)

The method of claim 18, wherein the composition is administered intramuscularly, intraocularly, stereotactically, intravenous, intraperitoneally or topically.

### Claim 22 (depends on 18)

The method of claim 18, wherein the mammal is a human.

### Claim 23 (depends on 18)

The method of claim 18, wherein the disease or physiological condition associated with mitochondrial dysfunction is selected from the group comprising of Myoclonic Epilepsy with Ragged Red Fibers (MERRF), Leigh Syndrome, Parkinson syndrome, Neuropathy and Myopathy, Multiple Symmetric Lipomatosis, Cardiomyopathy, Progressive External Opthalmoplegia (PEO) with monoclonus, Maternally Inherited Diabetes with Deafness (MIDD), Mitochondrial Encephalopathy, Lactic Acidosis and Stroke (MELAS), Riboflavin sensitive myopathy, Isolated Cardiomyopathy, Maternally Inherited Diabetes with Deafness (MIDD), Sudden Infant Death syndrome (SIDS), Maternal Deafness, Late Onset Alzheimer's Disease, Metabolic Syndrome, Neuropathy, Ataxia and Retinitis Pigmentosa (NARP), Leber's Hereditary Optic Neuropathy (LHON), Myopathy, Exercise Intolerance, Encephalopathy, Lactic Acidemia, Kearns-Sayre Syndrome (KSS), maternal or sporadic Syndromic Deafness, Pearson's Syndrome, Sideroblastic Anemia, Wolfram Syndrome (Diabetes Insipidus, Diabetes Mellitus, Optic Atrophy, Deafness or DIDMOAD), Myopathy and external opthalmoplegia; Neuropathy, Gastro-Intestinal and Encephalopathy (MNGIE), Inclusion Body Myositis, polymyositis with COX− muscle fibers and tissue injury.

### Claim 24 (depends on 13)

A method of preparing the composition as claimed in claim 13 comprising the steps of:
expressing coding sequences for subunits R6 (SEQ ID NOS: 1, 3, 5, 6, 7 and 8) and R8 (SEQ ID NOS: 1-8) in a prokaryotic strain and isolating the essentially pure mixture of subunits R6 and R8 thereby obtaining the carrier protein complex (component A) wherein the carrier protein is expressed in an E. coli strain and is purified;
preparing a recombinant polyribonucleotide segments each comprising a sequence set forth in SEQ ID NO:9, SEQ ID NO:10 or SEQ ID NO:11 by using a recombinant polynucleotide comprising a sequence set forth in SEQ ID NO:12 and a complementary strand thereof selected from the group consisting of SEQ ID NO:13 and SEQ ID NO:14 as starting material to obtain full-length SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11 (component B); and
mixing components A and B in the presence of a binding buffer and incubating the mixture for a time sufficient to form the composition containing a complex of components A and B.

### Claim 25

A diagnostic kit for screening mitochondrial dysfunction in a patient sample comprising:
a carrier (component A), wherein the carrier is subunit R6 (SEQ ID NOS: 1, 3, 5, 6, 7 and 8) and/or R8 (SEQ ID NOS: 1-8),
pc RNA (protein-coding RNA) (component B), wherein the pc RNA is a recombinant polyribonucleotide segments comprising a sequence set forth in SEQ ID NO:9, SEQ ID NO:10 and/or SEQ ID NO:11, a binding buffer and an instruction manual for carrying out diagnosis.