# US 11,998,618 B2 · Frataxin expression constructs > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,998,618 B2 - Document ID: US11998618B2 - Status: Granted - Kind: B2 - Application number: 17/125,449 - Filed: 2020-12-17 - Published: 2021-06-17 - Granted: 2024-06-04 - Earliest claimed: 2016-11-09 - Assignees: Precigen, Inc. - Applicants: Precigen, Inc. - Inventors: Stephen Schauer; Darby Thomas; Gregory Robinson; Mark Pykett; Richard Thorn; Kirsten Gruis - Web page: https://patentgrep.com/patent/US11998618B2 ## CPC - A61K 48/0058: {Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct} - A61K 38/1709: {from mammals} - A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered} - A61K 48/0066: {Manipulation of the nucleic acid to modify its expression pattern, e.g. enhance its duration of expression, achieved by the presence of particular introns in the delivered nucleic acid} - A61K 48/0075: {characterised by an aspect of the delivery route, e.g. oral, subcutaneous} - A61P 25/00: Drugs for disorders of the nervous system - C07K 14/47: from mammals - C12N 15/86: Viral vectors - C12N 15/8645: {Adeno-associated virus} - C12N 2750/14143: viral genome or elements thereof as genetic vector - C12N 2830/002: inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention provides polynucleotides, vectors and viruses expressing frataxin and methods of treating Friedreich's Ataxia. ## Claims ### Claim 1 A method of treating at least one symptom of Friedreich's Ataxia in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of a composition comprising: (a) an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to: a 5′UTR selected from 5U2, FTH1, GAPDH, and RPL6-5′Splice; a 3′UTR selected from a SV40 early adenylation sequence, a SV40 late adenylation sequence, a human growth hormone polyadenylation sequence (hGHpA), and a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; and a control element that directs transcription and/or translation; and (b) a pharmaceutically acceptable excipient, wherein the at least one symptom of Friedreich's Ataxia comprises loss of coordination in the arms and/or legs; and wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof. ### Claim 2 (depends on 1) The method of claim 1, wherein said frataxin is expressed in the mitochondria, cerebellum, hippocampus, anterior cortex, dorsal root ganglion, or a combination thereof of the subject. ### Claim 3 (depends on 1) The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 25% of normal levels of frataxin. ### Claim 4 (depends on 1) The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 30% of normal levels of frataxin. ### Claim 5 (depends on 1) The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 40% of normal levels of frataxin. ### Claim 6 (depends on 1) The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 50% of normal levels of frataxin. ### Claim 7 (depends on 1) The method of claim 1, wherein the AAV vector is an AAV5 vector. ### Claim 8 (depends on 1) The method of claim 1, wherein: the 5′UTR is FTH1 and the 3′UTR is a SV40 early polyadenylation sequence or a SV40 late polyadenylation sequence; the 5′UTR is GAPDH and the 3′UTR is a SV40 late polyadenylation sequence or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; the 5′UTR is 5U2 and the 3′UTR is a human growth hormone polyadenylation sequence (hGHpA) or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; or the 5′UTR is RPL6-5′Splice and the 3′UTR is a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7. ### Claim 9 (depends on 1) The method of claim 1, wherein the control element comprises a promoter. ### Claim 10 (depends on 9) The method of claim 9, wherein the promoter is a CMV promoter, a UBC promoter, an EF1α promoter, a PGK1 promoter, or a minimal frataxin promoter. ### Claim 11 (depends on 9) The method of claim 9, wherein the promoter is an inducible promoter. ### Claim 12 (depends on 11) The method of claim 11, wherein the AAV vector further comprises a gene switch that regulates the inducible promoter. ### Claim 13 (depends on 12) The method of claim 12, wherein the gene switch is an ecdysone receptor (EcR) gene switch. ### Claim 14 (depends on 1) The method of claim 1, wherein the pharmaceutically acceptable excipient comprises Na2HPO4, NaH2PO4, NaCl, KCl, CaCl2, MgCl2, or a combination thereof. ### Claim 15 (depends on 1) The method of claim 1, wherein the composition has a pH of about 6.5 to about 7.5. ### Claim 16 (depends on 1) The method of claim 1, wherein the composition comprises about 1×109 to about 2×1013 g/mL of the AAV vector. ### Claim 17 (depends on 1) The method of claim 1, wherein about 1×1010 to about 1×1015 vg of the AAV vector is administered to the subject. ### Claim 18 (depends on 1) The method of claim 1, wherein about 3.7×1010, about 1.11×1011, or about 3.7×1011 vg/g of the AAV vector is administered to the subject on a brain weight basis. ### Claim 19 A method of increasing frataxin levels in a subject having a frataxin deficiency, the method comprising administering to the subject: an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to: a 5′UTR selected from 5U2, FTH1, GAPDH, and RPL6-5′Splice; a 3′UTR selected from a SV40 early adenylation sequence, a SV40 late adenylation sequence, a human growth hormone polyadenylation sequence (hGHpA), and a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; and a control element that directs transcription and/or translation, wherein the frataxin encoded by the nucleic acid molecule is produced by the subject at a level that is at least 25% of frataxin levels produced by a subject not having a frataxin deficiency, wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof. ### Claim 20 A method of treating at least one symptom of Friedreich's Ataxia in a subject in need thereof, the method comprising administering to the subject a pharmaceutical formulation comprising: (a) an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to a 5′UTR, a 3′UTR, wherein: the 5′UTR is FTH1 and the 3′UTR is a SV40 early polyadenylation sequence or a SV40 late polyadenylation sequence; the 5′UTR is GAPDH and the 3′UTR is a SV40 late polyadenylation sequence or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; the 5′UTR is 5U2 and the 3′UTR is a human growth hormone polyadenylation sequence (hGHpA) or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; or the 5′UTR is RPL6-5′Splice and the 3′UTR is a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7, and (b) phosphate buffer solution (PBS) or Dulbecco's phosphate buffer solution (DPBS), wherein the frataxin encoded by the at least one nucleic acid molecule is produced by the subject at a level sufficient to treat at least one symptom of Friedreich's Ataxia in the subject; and wherein the at least one symptom of Friedreich's Ataxia comprises loss of coordination in the arms and/or legs; and wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof. ### Claim 21 (depends on 20) The method of claim 20, wherein about 1×1010 to about 1×1015 vg of the AAV vector is administered to the subject.