WO2024066780A1 - Nouveau virus adéno-associé de type fusion et son utilisation - Google Patents

Nouveau virus adéno-associé de type fusion et son utilisation Download PDF

Info

Publication number
WO2024066780A1
WO2024066780A1 PCT/CN2023/113302 CN2023113302W WO2024066780A1 WO 2024066780 A1 WO2024066780 A1 WO 2024066780A1 CN 2023113302 W CN2023113302 W CN 2023113302W WO 2024066780 A1 WO2024066780 A1 WO 2024066780A1
Authority
WO
WIPO (PCT)
Prior art keywords
adeno
fusion
associated virus
aav
disease
Prior art date
Application number
PCT/CN2023/113302
Other languages
English (en)
Chinese (zh)
Inventor
钟桂生
储岑凤
Original Assignee
上海玮美基因科技有限责任公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 上海玮美基因科技有限责任公司 filed Critical 上海玮美基因科技有限责任公司
Publication of WO2024066780A1 publication Critical patent/WO2024066780A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C07K14/01—DNA viruses
    • C07K14/015—Parvoviridae, e.g. feline panleukopenia virus, human parvovirus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C12N15/864—Parvoviral vectors, e.g. parvovirus, densovirus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof

Definitions

  • the present invention relates to the field of biomedicine, and in particular to a fusion adeno-associated virus and its application, and in particular to the application of heterologous gene delivery in the lungs or eyes.
  • Biological methods mainly refer to virus-mediated gene transfer, which uses viruses as vectors to recombinantly transfer heterologous genes with viruses through recombinant technology, infect receptor cells to express the target gene to treat diseases.
  • viral vector-mediated gene therapy has been widely used in clinical trials to treat cardiovascular, muscle, metabolic, nervous system, blood, sensory, and infectious diseases and cancer.
  • Adeno-associated virus belonging to the family of Parvoviridae, is a single-stranded DNA defective virus with the simplest structure found so far, and its replication process usually requires the participation of helper viruses.
  • Adeno-associated virus as a viral vector for the introduction of heterologous genes, has many significant advantages.
  • AAV is a non-pathogenic microorganism with high biosafety; second, AAV can infect not only dividing cells but also non-dividing cells, and its applicable host range is very wide; in addition, the genome of AAV is only about 5kb, which is convenient for recombination and operation; in addition, the physical properties of AAV are stable and easy to store, and high temperature of 56°C and pH fluctuations in the range of 3-9 do not affect its infection activity.
  • the above characteristics have led to the widespread use of AAV as a gene transfer vector, making it a hot spot in gene therapy research.
  • the lung is an important target organ for gene therapy, where the lung is prone to the most common genetic diseases, such as cystic fibrosis (CF) and ⁇ 1-antitrypsin deficiency ( ⁇ 1AT).
  • CF cystic fibrosis
  • ⁇ 1AT ⁇ 1-antitrypsin deficiency
  • lung cancer is also the most common cause of death in humans.
  • about 400 clinical programs for human gene therapy have been approved, of which about 10% are for lung diseases.
  • the data obtained from these human trials have successfully demonstrated that it is possible to transfer genes to human lungs, and the strategy of overexpressing heterologous genes to treat or control lung diseases has potential prospects. So far, the delivery of heterologous proteins or nucleic acids to the lungs through viral vectors is limited by the low infection efficiency of the current vectors, and effective infection cannot be formed under low-dose conditions.
  • the purpose of the present invention is to provide a fusion adeno-associated virus, which is used to solve the problems of low lung infection efficiency under low-dose infection conditions of AAV virus, high safety risks under high-dose infection, and difficulty in virus delivery in tissues such as the retina, for gene therapy.
  • the present invention infects the proximal and distal lungs of mice with the modified fusion-type new adeno-associated virus vector in vivo, which can mediate the specific expression of the target gene in lung cells under low-dose virus infection.
  • the new adeno-associated virus has broad application value and market prospects in the structural and functional analysis of lung cells, the establishment of disease models, and gene therapy.
  • the virus was injected into the mouse eyeball through the vitreous body and found that the virus vector has better infection characteristics than the current existing vectors, and also has broad application value and market prospects in the structural and functional analysis of ophthalmic cells, the establishment of disease models, and gene therapy.
  • the present invention provides a fusion adeno-associated virus AAV-WM03 capsid protein, which comprises a fusion peptide segment of serotypes AAV1 and AAV6 or a variant thereof.
  • the present invention also provides a nucleic acid, which encodes the capsid protein of the fusion adeno-associated virus AAV-WM03.
  • the present invention also provides a construct, which contains the nucleic acid.
  • the present invention also provides a host cell, wherein the host cell comprises the construct or the heterologous nucleic acid is integrated into the genome, or the host cell comprises the fusion adeno-associated virus AAV-WM03.
  • the present invention also provides a fusion-type adeno-associated virus AAV-WM03, the capsid structure of the fusion-type adeno-associated virus comprising any one of the fusion-type adeno-associated virus AAV-WM03 capsid proteins.
  • the present invention also provides a fusion adeno-associated virus vector system, comprising a packaging plasmid and an auxiliary plasmid.
  • the packaging plasmid comprises the nucleic acid, the nucleic acid fragment and the construct.
  • the present invention also provides a fusion-type adeno-associated virus, which is obtained by packaging the fusion-type adeno-associated virus vector system through viruses.
  • the present invention also provides a pharmaceutical composition, which comprises the fusion adeno-associated virus and pharmaceutically acceptable excipients.
  • the present invention also provides the use of the fusion adeno-associated virus AAV-WM03 capsid protein, nucleic acid, construct, fusion adeno-associated virus, host cell, fusion adeno-associated virus vector system, pharmaceutical composition or conjugate in the preparation of drugs for treating diseases; preferably, use in the preparation of drugs for gene therapy diseases; more preferably, use in the preparation of gene therapy drugs for lung diseases or eye diseases.
  • the fusion-type novel adeno-associated virus of the present invention has the following advantages and beneficial effects: (1) It provides a novel adeno-associated virus AAV-WM03 that can efficiently infect lung cells under low-dose conditions of AAV virus infection; (2) It provides a novel adeno-associated virus vector AAV-WM03 that can mediate the specific expression of the target gene in the RPE layer of the retina of adult mice; (3) The novel adeno-associated virus vector AAV-WM03 mediates the expression of the target gene more flexibly, with higher safety, more convenient application and lower cost than traditional transgenic methods; (4) The novel specific adeno-associated virus vector AAV-WM03 provides an application basis for the development of gene therapy for the precise treatment of lung and eye-related diseases.
  • FIG. 1a Schematic diagram of AAV-WM03 sequence.
  • Figure 1b Sequence alignment of AAV-WM03 and its parental vector.
  • Figure 1c Computer-simulated structure diagram of AAV-WM03.
  • FIG. 3a Immunofluorescence images of lungs infected with high doses of AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • Figure 3b Statistical graph and average infection status of proximal lung infection with high-dose AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • Figure 3c Statistical graph and average infection status of distal lung infection with high-dose AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • FIG. 4a Immunofluorescence images of lungs infected with low-dose AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • Figure 4b Statistical graph and average infection status of proximal lung infection with low-dose AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • Figure 4c Statistical graph and average infection status of distal lung infection with low-dose AAV-WM03, AAV6, AAV6.2ff, and AAV9.
  • the present invention provides a fusion adeno-associated virus AAV-WM03 capsid protein, which comprises a peptide segment (fusion peptide segment or chimeric peptide segment) formed by fusion of peptide segments of serotypes AAV1 and AAV6 or a variant thereof.
  • the fusion peptide segment or its variant comprises one or more segments of the peptide segments of AAV1 and 6.
  • the fusion peptide segment consists of two peptide segments, namely the first peptide segment and the second peptide segment.
  • the first peptide segment comprises a peptide segment from AAV1
  • the second peptide segment comprises a peptide segment from AAV6.
  • the first peptide segment and the second peptide segment are connected in sequence.
  • the first peptide segment comprises the amino acid fragment shown in SEQ ID No.1
  • the second peptide segment comprises the amino acid fragment shown in SEQ ID No.2.
  • the peptide segment of AAV1 contains one or more mutated amino acid sequence sites.
  • the peptide segment of AAV1 contains one mutation site.
  • the position and mutation type of the mutation site are S268R.
  • the peptide segments are directly fused to each other.
  • nucleocapsid of adeno-associated virus is generally nearly circular.
  • This nearly circular capsid is actually a closed icosahedral symmetrical hollow capsid composed of multiple protein capsomeres, in which the genomic nucleic acid is encapsidated.
  • An icosahedral symmetrical structure contains three types of rotational symmetry: 3-fold, 2-fold, and 5-fold symmetry (3-, 2-, 5-fold symmetry).
  • this symmetrical three-dimensional structure has a 3-fold symmetry axis passing through the center points of the two opposite faces of the virus particle, and the capsomeres rotate 120° around the 3-fold symmetry axis three times to reset, forming a triangular face; there is a 2-fold symmetry axis (edge), and the capsomeres rotate 180° around the 2-fold symmetry axis and reset twice to form two intersecting triangular faces; there is also a 5-fold symmetry axis passing through two opposite vertices, and the capsomeres rotate 72° around the 5-fold symmetry axis five times to reset, forming a pentamer. Therefore, the icosahedral symmetrical capsid is composed of 20 equilateral triangular faces, of which Every 2 triangles intersect to form an edge, with a total of 30 edges; every 5 triangles are connected to form 12 vertices.
  • the peptide segments of the serotypes AAV1 and AAV6 are assembled into the three-fold symmetry axis protrusions of the fusion adeno-associated virus AAV-WM03 capsid protein; the peptide segments of the serotypes AAV1 and AAV6 are assembled into the five-fold symmetry axis channels of the fusion adeno-associated virus AAV-WM03 capsid protein, and the computer simulated structure schematic diagram is shown in Figure 1c.
  • the fusion adeno-associated virus AAV-WM03 capsid protein comprises:
  • the polypeptide fragment in b) specifically refers to: a polypeptide fragment obtained by replacing, deleting or adding one or more (specifically 1-50, 1-30, 1-20, 1-10, 1-5 or 1-3) amino acids of the amino acid sequence as shown in SEQ ID No.4, or by adding one or more (specifically 1-50, 1-30, 1-20, 1-10, 1-5 or 1-3) amino acids at the N-terminus and/or C-terminus, and having the function of the polypeptide fragment as shown in the amino acid sequence as SEQ ID No.4, and the amino acid sequence in b) may have a sequence identity of 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or more than 99% with SEQ ID No.4.
  • the nucleotide sequence corresponding to the amino acid sequence of SEQ ID No. 4 includes one or more.
  • the nucleotide sequence is the nucleotide sequence shown in SEQ ID No. 3.
  • the present invention also provides a construct, which contains the above nucleotide sequence.
  • the construct is obtained by inserting the above nucleotide sequence into a suitable expression vector, and those skilled in the art can select a suitable expression vector.
  • the present invention also provides a host cell, wherein the host cell comprises the above-mentioned construct or the above-mentioned nucleotide sequence is integrated into the genome, or the host cell comprises the fusion-type adeno-associated virus AAV-WM03 as described in any one of the above items.
  • the host cell is selected from any one of mammalian cells (such as CHO, COS and N2A), plant cells, human cells (human cervical cancer cells such as HELA and human embryonic kidney cells such as HEK293T), bacterial cells (such as Escherichia coli, Streptomyces, Salmonella typhimurium), fungal cells (such as yeast), and insect cells (such as Sf9).
  • the host cell is an animal cell, and more preferably a human cell.
  • the host cell is a passage cell or a primary cell, i.e., a cell directly isolated from an organism (such as a human).
  • the host cell is an adherent cell or a suspended cell, i.e., a cell growing in the form of a suspension.
  • the present invention also provides a fusion-type adeno-associated virus AAV-WM03, wherein the fusion-type adeno-associated virus contains the fusion-type adeno-associated virus AAV-WM03 capsid protein as described in any one of the above items.
  • the fusion adeno-associated virus AAV-WM03 also includes a heterologous nucleotide sequence encoding a target product.
  • the heterologous nucleotide sequence encoding the target product can be wrapped and carried by various capsid proteins.
  • the heterologous nucleotide sequence encoding the target product is a construct, and the construct is a nucleic acid containing the target product.
  • the construct is constructed by inserting the nucleic acid encoding the target product into a suitable expression vector. Those skilled in the art can select a suitable expression vector.
  • the expression vector is selected from any one of pAAV-CAG, pAAV-TRE, pAAV-EF1a, pAAV-GFAP, pAAV-Lgr5, pAAV-Sox2, pAAV-Syn or pAAV-CMV expression vectors.
  • the target product is encoded as a nucleic acid or a protein
  • the nucleic acid is selected from a small guide RNA (sgRNA) and an interfering RNA (RNAi).
  • the sgRNA or RNAi targets a target selected from the epidermal growth factor receptor EGFR or the programmed death receptor PD-1.
  • the gene encoding the protein is selected from the D subtype surfactant protein SFTPD, the B subtype surfactant protein SFTPB, the A1 subtype surfactant protein SFTPA1, interferon ⁇ IFN- ⁇ , the cystic fibrosis transmembrane regulator CFTR or the aspartic protease Napsin A.
  • the present invention also provides an engineered cell transformed with the fusion-type adeno-associated virus as described above.
  • the engineered cell comprises the fusion-type adeno-associated virus as described above.
  • the cell is a eukaryotic cell and/or a prokaryotic cell.
  • any one of mammalian cells such as CHO, COS and N2A), plant cells, human cells (human cervical cancer cells such as HELA and human embryonic kidney cells such as HEK293FT), bacterial cells (such as Escherichia coli, Streptomyces, Salmonella typhimurium), fungal cells (such as yeast), and insect cells (such as Sf9) is selected.
  • the host cell is an animal cell, and more preferably a human cell.
  • the host cell is a passage cell or a primary cell, i.e., a cell directly isolated from an organism (such as a human).
  • the host cell is an adherent cell or a suspended cell, i.e., a cell grown in the form of a suspension.
  • the present invention also provides a fusion adeno-associated virus vector system, which comprises a packaging plasmid, an expression plasmid and a helper plasmid.
  • the packaging plasmid comprises the nucleic acid or nucleic acid fragment as described above.
  • the expression plasmid comprises a heterologous nucleotide sequence encoding a target product.
  • the packaging plasmid also contains a rep gene fragment of an adeno-associated virus, wherein the rep gene fragment contains an intron, and the intron contains a transcription termination sequence.
  • the target product is a nucleic acid or a protein
  • the nucleic acid is selected from small guide RNA (sgRNA) and interfering RNA (RNAi).
  • the sgRNA or RNAi targets a target selected from epidermal growth factor receptor (EGFR) and programmed death receptor 1 (PD-1).
  • the protein encoding gene is selected from surfactant protein subtype D (SFTPD), surfactant protein subtype B (SFTPB), surfactant protein subtype A1 (SFTPA1), interferon ⁇ IFN- ⁇ , cystic fibrosis transmembrane regulator (CFTR) and aspartic protease Napsin A.
  • the packaging plasmid, expression plasmid, and helper virus plasmid are transferred into host cells, and the nucleic acid sequences therein are all integrated into the host cells to produce the fusion adeno-associated virus.
  • the nucleic acid sequences encoding the various genes are present as separate expression cassettes, which prevent any risk of recombination to form a virus capable of replication; the nucleic acid sequences encoding the rep and cap genes are present in the same expression cassette.
  • the present invention also provides a fusion adeno-associated virus, which is obtained by virus packaging of the fusion adeno-associated virus vector system as described above.
  • the present invention also provides a pharmaceutical composition, comprising the fusion-type adeno-associated virus as described above and a pharmaceutically acceptable excipient.
  • the pharmaceutical composition or conjugate is administered by a systemic route or a local route, selected from inner ear administration, ophthalmic administration, intravenous administration, intramuscular administration, subcutaneous administration, oral administration, local contact, intraperitoneal administration and intralesional administration.
  • the pharmaceutical composition or conjugate dosage form is one or more of an injection, a tablet, a capsule, an aerosol, an eye drop or a nasal drop.
  • the adjuvant includes various excipients and diluents, which are not necessary active ingredients and have no excessive toxicity after application.
  • the adjuvant includes sterile water or saline, stabilizers, excipients, antioxidants (ascorbic acid, etc.), buffers (phosphoric acid, citric acid, other organic acids, etc.), preservatives, surfactants (PEG, Tween, etc.), chelating agents (EDTA, etc.) or adhesives.
  • the adjuvant also includes other low molecular weight polypeptides, serum albumin, glycine, glutamine, asparagine, arginine, polysaccharides, monosaccharides, mannitol or sorbitol.
  • the adjuvant is selected from saline, glucose isotonic solution, D-sorbitol isotonic solution, D-mannose isotonic solution, D-mannoses or sugar alcohol isotonic solution when used for the aqueous solution of injection.
  • the aqueous solution of injection includes a solubilizing agent.
  • the solubilizing agent is selected from alcohol (ethanol), polyol (propylene glycol or PEG) and/or nonionic surfactant (Tween 80 or HCO-50).
  • the fusion adeno-associated virus AAV-WM03 is a single active ingredient, and can also be combined with one or more other active components useful for treating lung diseases or eye diseases to form a combined preparation.
  • the active components are various other drugs for treating lung diseases or eye diseases.
  • the content of the active ingredient in the pharmaceutical composition is a safe and effective amount, which should be adjustable by those skilled in the art.
  • the dosage of the fusion adeno-associated virus AAV-WM03 and the active ingredient of the pharmaceutical composition depends on the patient's weight, the type of application, the condition and severity of the disease.
  • the dosage of the bifunctional compound as an active ingredient is 1-1000 mg/kg/day, 1-3 mg/kg/day, 3-5 mg/kg/day, 5-10 mg/kg/day, 10-20 mg/kg/day, 20-30 mg/kg/day, 30-40 mg/kg/day, 40-60 mg/kg/day, 60-80 mg/kg/day, 80-100 mg/kg/day, 100-200 mg/kg/day, 200-500 mg/kg/day, or greater than 500 mg/kg/day.
  • the present invention also provides a conjugate, which includes the fusion adeno-associated virus AAV-WM03 as described above or a connected biologically active polypeptide.
  • the pulmonary disease is selected from one or more of pneumonia, pulmonary fibrosis, and pulmonary tuberculosis.
  • the ophthalmic disease is also selected from age-related macular degeneration (AMD), choroidal neovascularization (CNV), choroidal neovascularization (CNV), Angiogenic membrane (CNVM), cystoid macular edema (CME), epiretinal membrane (ERM) and macular hole, angioid streaks, retinal detachment, diabetic retinopathy, diabetic macular edema (DME), atrophic lesions of retinal pigment epithelial cells (RPE), hypertrophic lesions of retinal pigment epithelial cells (RPE), retinal vein occlusion, chorioretinal vein occlusion, macular edema, pterygium conjunctiva, subretinal edema and intraretinal edema, retinitis pigmentosa, Stargardt's disease, glaucoma, inflammatory diseases, cataract, refractory abnormal phenomenon, keratoconus
  • the inflammation is selected from skin inflammation, vascular inflammation, allergy, autoimmune disease, fibrosis, scleroderma or transplant rejection; the autoimmune disease is selected from one or more of rheumatoid arthritis, systemic sclerosis, systemic lupus erythematosus, Sjögren's syndrome or polymyositis.
  • the cancer is selected from lymphoma, hematological tumor or solid tumor; specifically, the cancer is selected from adrenal cortical carcinoma, bladder urothelial carcinoma, breast cancer, cervical squamous cell carcinoma, endocervical adenocarcinoma, bile duct cancer, colon adenocarcinoma, lymphoid tumors, diffuse large B-cell lymphoma, esophageal cancer, glioblastoma multiforme, head and neck squamous cell carcinoma, renal chromophobe cell carcinoma, renal clear cell carcinoma, renal papillary cell carcinoma, acute myeloid leukemia, brain low-grade glioma, hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, mesothelial cell
  • the invention relates to a method for treating a leukemia or a diabetic renal cell carcinoma, wherein the leukemia or the diabetic renal cell carcinoma is selected from the group consisting of: a)
  • the metabolic disease is selected from diabetes, selected from type I and type II diabetes and diseases and conditions related to diabetes; the metabolic disease is selected from one or more of atherosclerosis, cardiovascular disease, nephropathy, neuropathy, retinopathy, ⁇ -cell dysfunction, dyslipidemia, hyperglycemia, insulin resistance or chronic obstructive pulmonary disease.
  • the gene therapy refers to the treatment of lung diseases.
  • the fusion adeno-associated virus or pharmaceutical composition can deliver the target product to lung cells under low-dose conditions of AAV virus infection to achieve the treatment of lung diseases.
  • the fusion adeno-associated virus AAV-WM03, host cell, vector system, pharmaceutical composition or conjugate of the present invention is used to deliver the target product to lung cells under low-dose conditions of AAV virus infection to achieve the treatment of lung diseases.
  • the delivery of the target product is for non-diagnostic treatment purposes, including delivery of the target product to lung cells in vitro.
  • the lung disease may be caused by pulmonary fibrosis caused by environmental factors. Therefore, the present invention also provides the use of the fusion adeno-associated virus in a drug for treating lung diseases caused by environmental factors in an individual.
  • the lung disease is a lung epithelial cell-related disease.
  • the lung epithelial Cell diseases can be prevented or treated by overexpressing Napsin A to inhibit cell phenotypic transformation and proliferation ability.
  • the lung disease is a disease related to gene defects, environmental damage or aging, for example, a disease related to gene mutation, etc., another example, a disease related to air pollution or drugs, and another example, a disease related to aging.
  • lung diseases are diseases related to cell damage, etc., specifically lung epithelial cell damage and/or fibroblast damage, more specifically lung epithelial cell damage caused by gene mutation, fibroblast damage caused by gene mutation, etc., cell damage caused by noise, cell damage caused by drugs, or cell damage caused by aging.
  • the fusion adeno-associated virus is used as a vector for delivering the target product.
  • the present invention also provides a method for treating a lung disease, comprising administering an effective amount of the fusion-type adeno-associated virus of the present invention, the host cell of the present invention, the vector system or the pharmaceutical composition or conjugate to a subject in need thereof.
  • a physician can determine the actual dosage that is most suitable for a single patient and it will vary according to the age, weight and response of a specific individual.
  • the fusion adeno-associated virus or host cell or vector system or pharmaceutical composition of the present invention can be administered to a patient.
  • a person skilled in the art can determine the appropriate administration method and dosage.
  • the delivery of one or more therapeutic genes by the fusion adeno-associated virus of the present invention can be used alone or in combination with other treatment methods or therapeutic components.
  • the fusion adeno-associated virus of the present invention is used to infect cells, thereby delivering genes and/or linked (e.g., but not limited to, covalently linked) biologically active polypeptides to cells. Therefore, the present invention provides a method for delivering heterologous genes to cells, the method infecting cells by allowing one or more fusion adeno-associated viruses or conjugates of the present invention to the cells, wherein the fusion adeno-associated virus or conjugate comprises one or more heterologous genes.
  • the present invention also provides a method for producing a stable fusion adeno-associated virus vector production cell line, comprising:
  • the AAV vector production cell is a mammalian cell.
  • the mammalian cell is selected from HEK293 cells, CHO cells, Jurkat cells, K562 cells, PerC6 cells, HeLa cells or derivatives thereof.
  • the mammalian host cell is a HEK293 cell, or is derived from a HEK293 cell.
  • the HEK293 cell is a HEK293T cell.
  • genomic sequences of various serotypes of AAV as well as the sequences of native ITRs, Rep proteins, and capsid proteins are known in the art. Such sequences can be found in the literature or in public databases such as GenBank. The disclosures thereof are incorporated herein by reference. The text is for AAV nucleic acid and amino acid sequences.
  • the active compound is co-administered with the other therapeutic agents.
  • Co-administered means that they are administered simultaneously in the same preparation or in two different preparations via the same or different routes, or sequentially via the same or different routes.
  • Sequential administration means that there is a time difference of seconds, minutes, hours or days between the administration of two or more different compounds.
  • the fusion adeno-associated viruses and methods of the invention can be used to prevent lung disease and can be administered as a preventive treatment prior to lung damage or after a period of time after exposure to an environment susceptible to lung damage.
  • vector refers to a macromolecule or combination of macromolecules that contains or is associated with a polypeptide and can be used to mediate delivery of the polypeptide to a cell.
  • Illustrative vectors include, for example, plasmids, viral vectors, liposomes or other gene delivery vectors.
  • AAV is an abbreviation for adeno-associated virus and may be used to refer to the virus itself or its derivatives.
  • recombinant AAV vector refers to an AAV vector containing a heterologous polynucleotide sequence, usually a sequence of interest for genetic transformation of cells.
  • heterologous polynucleotide is flanked by at least one, usually two, AAV inverted terminal repeats (ITRs).
  • AAV virus or "AAV viral particle” or “AAV vector particle” refers to a viral particle of an AAV vector comprising at least one AAV capsid protein and an encapsidated polynucleotide.
  • packing refers to the series of intracellular processes that lead to the assembly and encapsulation of AAV particles.
  • AAV rep and cap genes refer to polynucleotide sequences encoding the replication and packaging proteins of the adeno-associated virus.
  • AAV rep and cap refer to the AAV "packaging genes”.
  • helper virus of AAV refers to a virus that enables AAV to be replicated and packaged by mammalian cells.
  • AAV helper viruses are known in the art, including adenovirus, herpes virus, and pox virus (eg, vaccinia).
  • infectious virus or viral particle is one that contains a polynucleotide component capable of delivering the virus to cells tropistic for that virus species. The term does not necessarily imply that the virus has any ability to replicate.
  • producer cell refers to a cell line that has the AAV packaging genes (rep and cap genes), required helper viral genes, and the DNA genome of the recombinant AAV vector (e.g., the transgene of interest flanked by two AAV inverted terminal repeats (ITRs)) stably integrated into the host cell genome.
  • AAV packaging genes rep and cap genes
  • required helper viral genes e.g., the transgene of interest flanked by two AAV inverted terminal repeats (ITRs)
  • subject generally includes humans, non-human primates, such as mammals, dogs, cats, horses, sheep, pigs, cows, etc., who can benefit from treatment using the formulation, kit or combination formulation.
  • terapéuticaally effective amount generally refers to an amount that can achieve the effect of treating the diseases listed above after an appropriate administration period.
  • treatment and prevention are to be understood in their broadest sense.
  • the term “therapeutic” does not necessarily imply that the mammal is treated until full recovery.
  • prophylactic does not necessarily mean that the subject will not eventually contract a disease condition.
  • treatment and prevention include alleviating the symptoms of a particular condition or preventing or reducing the risk of a particular condition developing.
  • prevention may be understood to mean reducing the severity of an attack of a particular condition. Treatment may also reduce the severity of an existing condition or the frequency of acute attacks.
  • the subject or individual for therapeutic or preventive treatment is preferably a mammal, such as, but not limited to, a human, a primate, livestock (such as sheep, cattle, horses, donkeys, pigs), a pet (such as dogs, cats), a laboratory test animal (such as mice, rabbits, rats, guinea pigs, hamsters), or a captured wild animal (such as foxes, deer).
  • livestock such as sheep, cattle, horses, donkeys, pigs
  • a pet such as dogs, cats
  • a laboratory test animal such as mice, rabbits, rats, guinea pigs, hamsters
  • a captured wild animal such as foxes, deer.
  • the subject is preferably a primate.
  • the subject is most preferably a human.
  • Example 1 Acquisition of a novel adeno-associated virus AAV-WM03
  • the serotype of AAV is determined by the sequence of the Cap protein in the Rep-Cap plasmid required for AAV packaging. Therefore, AAVs produced by packaging with different Cap sequences have different infection tropisms.
  • the existing natural serotype was modified by fusing the amino acid sequence SEQ ID No.1 derived from AAV1 and the amino acid sequence SEQ ID No.2 derived from AAV6, wherein the sequence of AAV1 contains the mutation site S268R ( Figure 1a, b), together forming a new AAV serotype, named AAV-WM03, whose nucleotide sequence is SEQ ID No.3 and amino acid sequence is SEQ ID No.4.
  • Both the inner and outer surfaces of the capsid show the presence of capsid fragments from the two parents AAV1 and AAV6, and indicate that the three-fold symmetry axis protrusions and the five-fold symmetry axis channels of AAV-WM03 are also composed of AAV1 and 6 ( Figure 1c).
  • the corresponding nucleic acid sequence sources or combinations include multiple ones, among which a DNA sequence of the AAV-WM03 capsid protein can be translated, as shown in Figure 2, which is derived from AAV1, AAV3 and AAV13, and contains three base mutations of AAV3, corresponding to the S268R amino acid mutation ( Figure 2), that is, the nucleic acid sequence of the fusion adeno-associated virus AAV-WM03, which is fused from serotypes AAV1 and AAV6, can also be composed of serotypes AAV1, AAV3 and AAV13.
  • the sequence of the coding gene of AAV-WM03 obtained in the above sequencing results was synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. to obtain the Rep-Cap plasmid of AAV-WM03, namely pAAV-WM03.
  • the obtained Rep-Cap plasmid pAAV-WM03 of AAV-WM03, the genomic plasmid pAAV-CMV-EGFP expressing a green fluorescent protein EGFP, the nucleotide sequence SEQ ID No.5, and the pHelper plasmid (the full sequence of the plasmid is the same as that of SEQ ID NO.12 of the AAV-ie patent document CN110437317A) were co-transfected into HEK-293T cells at a plasmid dosage of 80, 60, and 110 ⁇ g per plate of cells, and the AAV virus was purified by iodide dialkyl gradient ultracentrifugation. The virus titer was measured at 2E+13GC/mL as the appropriate concentration, and it was placed at -80°C for use.
  • Example 2 In vivo verification of the novel adeno-associated virus AAV-WM03-mNeonGreen in the lungs
  • AAV6 and AAV6.2ff are two AAV vectors known to be effective against lung infections.
  • the experiment set up four test viruses, AAV6, AAV9, AAV6.2ff, and AAV-WM03, which were packaged with green fluorescent protein mNeonGreen, and set up high-dose groups (5E+10GC/mL) and low-dose groups (2.5E+10GC/mL), for a total of 8 groups. All test groups used 4 mice aged 6-8 weeks for intratracheal injection into the lungs. Anesthetize the anesthetized mice and fix them on the animal workbench in a dorsal position, straighten their front paws to both sides of the body and fix them with tape.
  • the infection efficiency of AAV-WM03 in the proximal part of the lungs under high dose conditions is 59.67 ⁇ 1.202
  • the infection efficiency of AAV6 in the proximal part of the lungs used as a comparison with AAV-WM03 is 39.00 ⁇ 1.528
  • AAV6.2ff is 77.67 ⁇ 1.453
  • a AV9 was 83.00 ⁇ 2.887
  • the infection efficiency of AAV-WM03 in the distal lung was 54.33 ⁇ 0.667
  • the infection efficiency of AAV6 in the distal lung, which was used as a comparison with AAV-WM03 was 27.67 ⁇ 1.453
  • AAV6.2ff was 64.33 ⁇ 1.202
  • AAV9 was 72.67 ⁇ 5.783
  • AAV-WM03 had an advantage over natural serotype AAV6 and its mutant AAV6.2ff in infecting lung tissues in the proximal and distal parts of the lungs ( Figure 4a).
  • the infection efficiency of AAV-WM03 in the proximal part of the lungs was 61.00 ⁇ 1.528, while the infection efficiency of AAV6, which was used as a comparison with AAV-WM03, in the proximal part of the lungs was 59.67 ⁇ 0.882, AAV6.2ff was 41.00 ⁇ 1.155, and AAV9 was 6.
  • Example 3 In vivo verification of the novel adeno-associated virus AAV-WM03 in the visual system
  • the glass needle stays for 30s and then slowly withdraws.
  • the mouse After applying erythromycin ointment on the wound, the mouse is placed in a mouse cage in a 41°C water bath to keep warm. After the mouse wakes up, it is moved to the mouse room for breeding. After 11 days, the eye cup is taken. The mouse is killed by spinal dislocation. After removing the eyeball, the cornea is punctured with a 1ml syringe to release the aqueous humor. The eyeball is placed in a small culture dish filled with PBS solution and dissected under a microscope.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biomedical Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Molecular Biology (AREA)
  • Virology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Epidemiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Hematology (AREA)
  • Neurology (AREA)
  • Pulmonology (AREA)
  • Obesity (AREA)
  • Diabetes (AREA)
  • Ophthalmology & Optometry (AREA)
  • Neurosurgery (AREA)
  • Cell Biology (AREA)

Abstract

L'invention concerne un virus adéno-associé de type fusion et un effet d'administration de gène de celui-ci dans le poumon et les yeux. Le virus adéno-associé de type fusion comprend un fragment de peptide de fusion formé par fusion de fragments peptidiques de sérotype VAA1 et VAA6 ou d'un variant de celui-ci, infectant efficacement le poumon à une faible dose. Le virus adéno-associé peut également infecter efficacement une couche d'épithélium pigmentaire rétinien (RPE) pour l'administration de gènes aux yeux. Le virus adéno-associé de type fusion est utilisé en tant que support sûr, et a une large perspective d'application dans le traitement d'une maladie pulmonaire ou oculaire.
PCT/CN2023/113302 2022-09-30 2023-08-16 Nouveau virus adéno-associé de type fusion et son utilisation WO2024066780A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202211212092.0 2022-09-30
CN202211212092.0A CN117801116A (zh) 2022-09-30 2022-09-30 一种融合型新型腺相关病毒及其应用

Publications (1)

Publication Number Publication Date
WO2024066780A1 true WO2024066780A1 (fr) 2024-04-04

Family

ID=90420706

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2023/113302 WO2024066780A1 (fr) 2022-09-30 2023-08-16 Nouveau virus adéno-associé de type fusion et son utilisation

Country Status (2)

Country Link
CN (1) CN117801116A (fr)
WO (1) WO2024066780A1 (fr)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112218880A (zh) * 2018-03-29 2021-01-12 阿斯克肋匹奥生物制药公司 逃避中和的肝趋向性重组aav6载体
US20210017542A1 (en) * 2018-03-29 2021-01-21 University Of Florida Research Foundation, Incorporated Aav6 variants
CN112533644A (zh) * 2018-04-03 2021-03-19 斯特里迪比奥公司 靶向眼组织的病毒载体
US20210238260A1 (en) * 2020-01-29 2021-08-05 Catalyst Biosciences, Inc. Gene therapy for hemophilia b with a chimeric aav capsid vector encoding modified factor ix polypeptides
CN113544278A (zh) * 2019-01-28 2021-10-22 杜克大学 用于逃避体液免疫的组合物和方法
WO2022036220A1 (fr) * 2020-08-14 2022-02-17 The Trustees Of The University Of Pennsylvania Nouvelles capsides d'aav et compositions les contenant
CN116925192A (zh) * 2022-04-12 2023-10-24 合肥星眸生物科技有限公司 一种融合型腺相关病毒及其应用

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112218880A (zh) * 2018-03-29 2021-01-12 阿斯克肋匹奥生物制药公司 逃避中和的肝趋向性重组aav6载体
US20210017542A1 (en) * 2018-03-29 2021-01-21 University Of Florida Research Foundation, Incorporated Aav6 variants
CN112533644A (zh) * 2018-04-03 2021-03-19 斯特里迪比奥公司 靶向眼组织的病毒载体
CN113544278A (zh) * 2019-01-28 2021-10-22 杜克大学 用于逃避体液免疫的组合物和方法
US20210238260A1 (en) * 2020-01-29 2021-08-05 Catalyst Biosciences, Inc. Gene therapy for hemophilia b with a chimeric aav capsid vector encoding modified factor ix polypeptides
WO2022036220A1 (fr) * 2020-08-14 2022-02-17 The Trustees Of The University Of Pennsylvania Nouvelles capsides d'aav et compositions les contenant
CN116925192A (zh) * 2022-04-12 2023-10-24 合肥星眸生物科技有限公司 一种融合型腺相关病毒及其应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE Protein 26 July 2016 (2016-07-26), H ANONYMOUS: "capsid protein VP1, partial (endogenous virus) [Adeno-associated virus]", XP093154717, retrieved from NCBI Database accession no. ACB55303.1 *
KE JUNZI: "Advances in AAV mediated gene therapy for hearing loss", CHINESE JOURNAL OF NATURE, 上海科技大学IHUMAN研究所,上海201210, vol. 43, no. 1, 5 February 2021 (2021-02-05), pages 39 - 44, XP093154720, ISSN: 0253-9608, DOI: 10.3969/j.issn.0253-9608.2021.01.006 *
YAN, Z. ET AL.: "Distinct Transduction Difference Between Adeno-Associated Virus Type 1 and Type 6 Vectors in Human Polarized Airway Epithelia", GENE THERAPY, vol. 20, 14 June 2012 (2012-06-14), XP037772047, DOI: 10.1038/gt.2012.46 *

Also Published As

Publication number Publication date
CN117801116A (zh) 2024-04-02

Similar Documents

Publication Publication Date Title
WO2021179861A1 (fr) Vecteur de virus adéno-associé et son utilisation
JP2021097675A (ja) 中枢神経系を標的化したａａｖベクター
EP3471780B1 (fr) Traitement de la dmla à l'aide d'un variant d'aav2 et d'aflibercept
JP6449175B2 (ja) 眼の遺伝子関連疾患の治療のための方法及び組成物
JP2022084809A (ja) 滲出型加齢性黄斑変性の治療のための組成物
CN114848850B (zh) Rs1基因在制备xlrs治疗剂中的应用及治疗剂
JP2021530239A (ja) 遺伝子治療発現ベクターの眼内送達
WO2020215968A1 (fr) Vecteur aav pour infecter efficacement des cellules de soutien et des cellules ciliées
WO2020088548A1 (fr) Vecteurs de thérapie génique pour le traitement de la rétinite pigmentaire
WO2023198050A1 (fr) Virus adéno-associé de type fusion et son utilisation
EP3897679A1 (fr) Virus adéno-associés et leurs utilisations pour la thérapie de l'oreille interne
WO2020010491A1 (fr) Acide nucléique servant à coder pour une protéine du sous-motif 4 de la nadh déshydrogénase humaine et application associée
WO2024066780A1 (fr) Nouveau virus adéno-associé de type fusion et son utilisation
US20220348957A1 (en) Adeno-associated viruses and their uses for inner ear therapy
WO2020223933A1 (fr) Mutant d'aav qui infecte efficacement des cellules
WO2023207792A1 (fr) Nouvelle souche modifiée par capside de vaa et son utilisation
WO2024002076A1 (fr) Médicament aav pour traiter des maladies du fond de l'œil associées à l'angiogenèse
WO2023143366A1 (fr) Variant de virus adéno-associé et son application dans le traitement de maladies
WO2021195876A1 (fr) Vecteur d'expression du facteur 2 lié au facteur nucléaire humain e2 et application du vecteur d'expression
WO2023078099A1 (fr) Agent thérapeutique génétique pour le traitement des lésions nerveuses
WO2023236964A1 (fr) Composition pharmaceutique de virus adéno-associé et utilisation associée
EP4333903A1 (fr) Compositions et méthodes associées au traitement de maladies oculaires chez les équidés
WO2023014892A1 (fr) Administration intraoculaire durable médiée par des cellules oculaires d'agents thérapeutiques cellulaires pour le traitement de maladies ou de troubles oculaires
CN117866970A (zh) 用于无脉络膜症的基因工程构建体
NZ787256A (en) Compositions For Treatment of Wet Age-Related Macular Degeneration

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23870026

Country of ref document: EP

Kind code of ref document: A1