# US 2020/0040360 A1 · Genetic construct > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0040360 A1 - Document ID: US20200040360A1 - Status: Published application - Kind: A1 - Application number: 16/436,676 - Filed: 2019-06-10 - Published: 2020-02-06 - Earliest claimed: 2015-10-26 - Assignees: Quethera Limited - Applicants: Quethera Limited - Inventors: Peter WIDDOWSON; Keith MARTIN - Web page: https://patentgrep.com/patent/US20200040360A1 ## CPC - 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 38/185: {Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3} - A61K 48/0058: {Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct} - A61P 9/14: Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers - A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like - A61P 25/00: Drugs for disorders of the nervous system - A61P 25/02: for peripheral neuropathies - A61P 27/00: Drugs for disorders of the senses - A61P 27/02: Ophthalmic agents - A61P 27/06: Antiglaucoma agents or miotics - A61P 27/16: Otologicals - C07K 14/48: Nerve growth factor [NGF] - C12N 9/12: transferring phosphorus containing groups, e.g. kinases (2.7) - C12N 15/1027: {by DNA shuffling, e.g. RSR, STEP, RPR} - C12N 15/62: DNA sequences coding for fusion proteins - C12N 15/85: for animal cells - C12N 15/86: Viral vectors - C12N 15/8645: {Adeno-associated virus} - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy - C12N 2750/14132: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent - C12N 2750/14143: viral genome or elements thereof as genetic vector - C12N 2830/008: cell type or tissue specific enhancer/promoter combination - C12N 2840/203: having an IRES - 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 genetic constructs and recombinant vectors comprising such constructs. The constructs and vectors can be used in gene therapy methods for treating a range of disorders, including glaucoma and deafness, or for promoting nerve regeneration and/or survival. ## Claims ### Claim 1 A genetic construct comprising a promoter operably linked to a first coding sequence, which encodes the tyrosine kinase receptor B (TrkB), and a second coding sequence, which encodes an agonist of the TrkB receptor. ### Claim 2 (canceled) ### Claim 3 (canceled) ### Claim 4 (depends on 1) A genetic construct according to claim 1, wherein the promoter is the human synapsin I (SYN I) promoter or the CAG promoter. ### Claim 5 (canceled) ### Claim 6 (depends on 1) A genetic construct according to claim 1, wherein the genetic construct comprises a spacer sequence disposed between the first and second coding sequences, which spacer sequence encodes a peptide spacer that is configured to be digested to thereby produce the TrkB receptor and agonist as separate molecules. ### Claim 7 (depends on 6) A genetic construct according to claim 6, wherein the spacer sequence comprises and encodes a viral peptide spacer sequence, more preferably a viral 2A peptide spacer sequence. ### Claim 8 (depends on 6) A genetic construct according to claim 6, wherein the peptide spacer sequence comprises an amino acid sequence substantially as set out in SEQ ID NO. 4, or a fragment or variant thereof; or (ii) an amino acid sequence substantially as set out in SEQ ID NO. 6, or a fragment or variant thereof; or (iii) an amino acid sequence substantially as set out in SEQ ID NO. 8, or a fragment or variant thereof. ### Claim 9 (depends on 6) A genetic construct according to claim 6, wherein the spacer sequence comprises: (i) a nucleotide sequence substantially as set out in SEQ ID NO.5, or a fragment or variant thereof; or (ii) a nucleotide sequence substantially as set out in SEQ ID NO. 7, or a fragment or variant thereof. ### Claim 10 (canceled) ### Claim 11 (canceled) ### Claim 12 (canceled) ### Claim 13 (depends on 1) A genetic construct according to claim 1, wherein the first coding sequence comprises a nucleotide sequence encoding the human canonical isoform of TrkB comprising an amino acid sequence substantially as set out in SEQ ID NO. 9, or a fragment or variant thereof, and/or wherein the first coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 10, or a fragment or variant thereof. ### Claim 14 (canceled) ### Claim 15 (depends on 1) A genetic construct according to claim 1, wherein the first coding sequence comprises a nucleotide sequence which encodes isoform 4 of TrkB, and (i) wherein isoform 4 of TrkB comprises an amino acid sequence substantially as set out in SEQ ID NO. 11, or a fragment or variant thereof, and/or (ii) wherein the first coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 12, or a fragment or variant thereof. ### Claim 16 (canceled) ### Claim 17 (canceled) ### Claim 18 (depends on 1) A genetic construct according to claim 1, wherein the first coding sequence comprises a nucleotide sequence encoding a mutant form of TrkB receptor, wherein one or more tyrosine residue at position 516, 701, 705, 706 and/or 816 of SEQ ID No: 9 is modified or mutated and/or wherein at least two, three or four tyrosine residues at position 516, 701, 705, 706 and/or 816 of SEQ ID No: 9 are modified, and/or wherein all five tyrosine residues at position 516, 701, 705, 706 and/or 816 of SEQ ID No: 9 are modified, and/or wherein the or each tyrosine residue is modified to a glutamic acid. ### Claim 19 (canceled) ### Claim 20 (canceled) ### Claim 21 (canceled) ### Claim 22 (depends on 18) A genetic construct according to claim 18, wherein the modified form of the TrkB receptor comprises an amino acid sequence substantially as set out in SEQ ID NO. 13, or a fragment or variant thereof, and/or wherein the first coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 14, or a fragment or variant thereof. ### Claim 23 (canceled) ### Claim 24 (depends on 1) A genetic construct according to claim 1, wherein the second coding sequence encodes an agonist of the TrkB receptor, which is a member of the neurotrophin family of trophic factors, and wherein the agonist is selected from a group of agonists consisting of: Brain-derived neurotrophic factor (BDNF); nerve growth factor (NGF); neurotrophin-3 (NT-3); neurotrophin-4 (NT-4); and neurotrophin-5 (NT-5); or fragments thereof. ### Claim 25 (depends on 24) A genetic construct according to claim 24, wherein the second coding sequence encodes neurotrophin-4 (NT-4), which comprises an amino acid sequence substantially as set out in SEQ ID NO: 49 or 55, or a fragment or variant thereof, and/or the second coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID No: 50 or 56, or a fragment or variant thereof. ### Claim 26 (depends on 1) A genetic construct according to claim 1, wherein the agonist of the TrkB receptor is prepro-brain derived neurotrophic factor (pre-pro-BDNF), pro-BDNF or mature BDNF (mBDNF), wherein the proBDNF is canonical proBDNF, optionally wherein canonical proBDNF comprises an amino acid sequence substantially as set out in SEQ ID NO. 15, or a fragment or variant thereof, or wherein the second coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 16, or a fragment or variant thereof, and/or wherein the proBDNF is isoform 2 of proBDNF, optionally wherein isoform 2 of proBDNF comprises an amino acid sequence referred to herein as SEQ ID NO. 17, or a fragment or variant thereof. ### Claim 27 (canceled) ### Claim 28 (canceled) ### Claim 29 (depends on 26) A genetic construct according to claim 26, wherein the second coding sequence comprises a nucleotide sequence which encodes mature BDNF, and/or wherein mature BDNF comprises an amino acid sequence substantially as set out in SEQ ID NO. 18, or a fragment or variant thereof, and/or wherein the second coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 19, or a fragment or variant thereof. ### Claim 30 (canceled) ### Claim 31 (canceled) ### Claim 32 (depends on 1) A genetic construct according to claim 1, wherein the second coding sequence comprises a nucleotide sequence encoding a signal peptide for the agonist of the TrkB receptor, preferably a signal peptide for BDNF, optionally wherein the nucleotide sequence encodes the canonical signal peptide for BDNF. ### Claim 33 (canceled) ### Claim 34 (depends on 32) A genetic construct according to claim 32, wherein the second coding sequence comprises a nucleotide sequence which encodes a signal peptide comprising an amino acid sequence substantially as set out in SEQ ID NO. 20, or a fragment or variant thereof, and/or wherein the second coding sequence comprises a nucleotide sequence substantially as set out in SEQ ID NO. 21, or a fragment or variant thereof. ### Claim 35 (canceled) ### Claim 36 (depends on 1) A genetic construct according to claim 1, wherein the second coding sequence comprises a nucleotide sequence encoding a signal sequence peptide substantially as set out in any one of SEQ ID NO. 23, 25, 27 or 29, or wherein the signal peptide comprises an amino acid sequence substantially as set out in any one of SEQ ID NO. 22, 24, 26 or 28, and/or wherein the second coding sequence comprises a nucleotide sequence encoding a signal sequence peptide substantially as set out in any one of SEQ ID NO. 31, 33, 35, 37, 39, 41, 43, 45, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 or 103; or wherein the signal peptide comprises an amino acid sequence substantially as set out in any one of SEQ ID NO. 30, 32, 34, 36, 38, 40, 42, 44, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 or 102. ### Claim 37 (canceled) ### Claim 38 (depends on 1) A genetic construct according to claim 1, wherein the construct comprises a nucleotide sequence substantially as set out in SEQ ID No: 107 or 108, or a fragment or variant thereof. ### Claim 39 (depends on 1) A recombinant vector comprising the genetic construct according to claim 1, wherein the vector is a recombinant AAV (rAAV) vector, optionally wherein the rAAV is AAV-1, AAV-2, AAV-3A, AAV-3B, AAV-4, AAV-5, AAV-6, AAV-7, AAV-8, AAV-9, AAV-10 or AAV-11. ### Claim 40 (canceled) ### Claim 41 (canceled) ### Claim 42 (canceled) ### Claim 43 (canceled) ### Claim 44 (canceled) ### Claim 45 (canceled) ### Claim 46 (canceled) ### Claim 47 (canceled) ### Claim 48 (depends on 1) A pharmaceutical composition comprising the genetic construct according to claim 1, and a pharmaceutically acceptable vehicle. ### Claim 49 (canceled) ### Claim 50 (depends on 1) A method of treating, preventing or ameliorating an optic nerve disorder or a cochlear disorder in a subject, or for promoting nerve regeneration and/or survival in a subject, the method comprising administering, to a subject in need of such treatment, a therapeutically effective amount of the genetic construct according to claim 1. ### Claim 51 (depends on 50) The method according to claim 50, wherein the optic nerve disorder that is treated is any pathophysiological condition which results in loss of RGCs, such as trauma to the head or face or vascular insults, for example partial or complete loss in blood supply to the ocular structures or regions of the brain which receive input from the optic nerve, or is used to support replacement of RGCs through introduction of untransformed or transformed stem cell into the eye or regions associated with vision in patients, optionally wherein the optic nerve disorder that is treated is glaucoma, or wherein the cochlear disorder which is treated is hearing loss or deafness.