Klotho mrna
AssigneeAdvantage Therapeutics, Inc.
InventorsThomas Hoffmann, Agustin Fernandez Santana, III, Walter Schmidt
Life of this patent
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Abstract
The present invention relates to a Klotho messenger-RNA (mRNA), wherein the mRNA has a 5′ CAP region, a 5′ un-translated region (5′-UTR), a coding region encoding a Klotho polypeptide, a 3′ untranslated region (3′-UTR) and a poly-adenosine Tail (poly-A tail), wherein the Klotho polypeptide comprises the KL1 domain of human Klotho, preferably wherein the coding region encoding the Klotho polypeptide comprises an RNA sequence having at least 80% sequence identity to SEQ ID NO: 1, and wherein the coding region encoding the Klotho polypeptide preferably has a GC content of at least 54%.
Claims24
Klotho messenger-RNA (mRNA), wherein the mRNA has a 5′ CAP region, a 5′ untranslated region (5′-UTR), a coding region encoding a Klotho polypeptide, a 3′ untranslated region (3′-UTR) and a polyadenosine tail (poly-A tail),
- wherein the Klotho polypeptide comprises the KL1 domain of human Klotho,
- wherein the coding region encoding the Klotho polypeptide comprises an RNA sequence having at least 80% sequence identity to SEQ ID NO:
- 1 or 79, and
- wherein the coding region encoding the Klotho polypeptide has a GC content of at least 54%.
The Klotho mRNA according to any one of the preceding claims, wherein the coding region encoding the Klotho polypeptide has at least 80% sequence identity to SEQ ID NO:
- 18, 96, 41, 112, 68, or 124.
The Klotho mRNA according to any one of the preceding claims, wherein the coding region encoding the Klotho polypeptide has at least 93% sequence identity to at least one of SEQ ID NO:
- 1, 79, 13, 91, 18, 96, 41, 112, 68, or 124, and wherein the coding region encoding the Klotho polypeptide has a GC content between 54% and 66%.
The Klotho mRNA according to any one of the preceding claims, wherein the coding region encoding the Klotho polypeptide has at least 93% sequence identity to SEQ ID NO:
- 18 or 96, wherein the coding region encoding the Klotho polypeptide has a GC content between 54% and 66%, and wherein the Klotho polypeptide is SEQ ID NO:
- 57.
The Klotho mRNA according to any one of the preceding claims, wherein the coding region encoding the Klotho polypeptide comprises a sequence selected from SEQ ID NO:
- 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 64, 65, 66, 67, 68, 69, 70, 71, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 120, 121, 122, 123, 124, 125, 126, 127.
The Klotho mRNA according to any one of the preceding claims, wherein the coding region encoding the Klotho polypeptide is selected from SEQ ID NO:
- 18, 19, 20, 21, 96, 97, 98, 99, 41, 42, 43, 44, 112, 113, 114, 115, 68, 69, 70, 71, 124, 125, 126, 127.
The Klotho mRNA according to any one of the preceding claims, wherein the Klotho polypeptide has at least 90% sequence identity to SEQ ID NO:
- 49, 55, 57, or 59, preferably wherein the Klotho polypeptide is selected from SEQ ID NO:
- 49, 55, 56, 57, 58, 59, 60, or 76, preferably 49, 57 or 59.
The Klotho mRNA according to any one of the preceding claims, wherein the Klotho mRNA is SEQ ID NO:
- 29, 30, 31, 32, 100, 101, 102, 103, 45, 46, 47, 48, 116, 117, 118, 119, 72, 73, 74, 75, 128, 129, 130, 131;
- preferably SEQ ID NO:
- 29, 100, 45, 116, 72, 128.
The Klotho mRNA according to any one of the preceding claims, wherein in the Klotho mRNA, at least 5%, preferably at least 10%, preferably at least 30%, especially at least 50% of all
- cytidine residues are replaced by 5-methyl-cytidine residues, and/or
- cytidine residues are replaced by 2-amino-2-deoxy-cytidine residues, and/or
- cytidine residues are replaced by 2-fluoro-2-deoxy-cytidine residues, and/or
- cytidine residues are replaced by 2-thio-cytidine residues, and/or
- cytidine residues are replaced by 5-iodo-cytidine residues, and/or
- uridine residues are replaced by pseudo-uridine residues, and/or
- uridine residues are replaced by 1-methyl-pseudo-uridine residues, and/or
- uridine residues are replaced by 2-thio-uridine residues, and/or
- uridine residues are replaced by 5-methyl-uridine residues, and/or
- adenosine residues are replaced by N6-methyl-adenosine residues.
Pharmaceutical formulation comprising the Klotho mRNA according to any one of the preceding claims and a pharmaceutically acceptable carrier.
The pharmaceutical formulation according to the preceding claim, wherein the formulation comprises a further mRNA, wherein the further mRNA has a further 5′ CAP region, a further 5′-UTR, a further coding region encoding a further polypeptide, a further 3′-UTR and a further poly-A tail;
- preferably wherein the further polypeptide is Bactericidal/Permeability Increasing Protein Family B, member 4 (BPIFB4) protein or an isoform or variant thereof, preferably Longevity-Associated Variant of BPIFB4 (LAV-BPIFB4)
The pharmaceutical formulation according to any one of the preceding claims, wherein the further polypeptide is a protein selected from Apolipoprotein E, Tumor protein p53, Sirtuin 1 protein, FOXO1 transcription factor, Cholinergic receptor nicotinic alpha 3 subunit, SH2B adaptor protein 3, Cyclin dependent kinase inhibitor 2A, Elongation of very-long-chain fatty acids-like 2, Werner protein, Paraoxonase 1, Superoxide dismutase 2, Lamin A protein, Cholesteryl ester transfer protein, Apolipoprotein C3, Microsomal triglyceride transfer protein, Phosphatidylinositol 3-kinase (PI3K), Insulin-like growth factor 1 (IGF-1) receptor, Protein-L-isoaspartyl methyltransferase, Growth hormone, CAMP-response element binding protein, Mitogen-activated protein kinase, epidermal growth factor receptor, Nuclear factor kappa B, Phospholipase C beta, Methionine sulfoxide reductase A, Mediator of cell motility 1, Nei like DNA Glycosylase 1, Peroxisome proliferator-activated receptor gamma 2, Eukaryotic translation initiation factor 3 subunit K, ATM serine/threonine kinase, B-cell lymphoma 2, Cell division cycle 42, Diacylglycerol O-acyltransferase 1, Early growth response 1, Fibroblast growth factor 23, Fibroblast growth factor 21, Fructosamine 3 kinase related protein, Phosphoglycolate phosphatase, Insulin receptor substrate 1, Polycomb complex protein BMI-1, Neuregulin 1, Signal transducer and activator of transcription, E2F Transcription Factor 1, Vascular endothelial growth factor A, Xenobiotic metabolizing enzymes, Myc proto-oncogene protein, C—X—C chemokine receptor type 4, Silent information regulator 2, Extracellular signal-regulated kinase, and SLC31.
Kit for administering the Klotho mRNA according to any one of the preceding claims to an individual, preferably a human, comprising
- the Klotho mRNA according to any one of the preceding claims, and
- a device for administering the Klotho mRNA.
A DNA molecule comprising a DNA sequence encoding the coding region of the Klotho mRNA as defined in any one of the preceding claims.
A cell comprising the DNA molecule as defined in the preceding claim, preferably wherein the cell is a mammalian cell, preferably a Human Embryonic Kidney (HEK) 293 cell.
A therapeutic delivery device comprising cells as defined in the preceding claim, wherein the cells are encapsulated in a biocompatible scaffold.
An extracellular vesicle (EV) comprising the Klotho mRNA as defined in any one of the preceding claims, preferably wherein the EV is derived from the cell as defined in any one of the preceding claims.
A population of EVs as defined in the preceding claim, wherein the average number of Klotho mRNA molecules per EV throughout the population of EVs is above one per EV.
A method for producing the EV or the population of EVs according to any one of the preceding claims comprising:
- Culturing the cell as defined in any one of the preceding claims under conditions suitable for the formation of EVs;
- and
- Isolating the EVs.
A pharmaceutical formulation comprising the EV or the population of EVs as defined in any one of the preceding claims and a pharmaceutically acceptable carrier.
The Klotho mRNA, the pharmaceutical formulation, the therapeutic delivery device, the EV, or the population of EVs according to any one of the preceding claims for use as a medicament.
The Klotho mRNA, the pharmaceutical formulation, the therapeutic delivery device, the EV, or the population of EVs according to any one of the preceding claims for use in the prevention or treatment of an aging-related disorder and/or a symptom of aging.
Use of the Klotho mRNA, the EV, or the population of EVs according to any one of the preceding claims for increasing the lifespan in a mammal, preferably a human.
Dependencies are inferred from the claim text.