# US 2014/0155475 A1 · Modified polynucleotides for treating lipase a, lysosomal acid, cholesterol esterase protein deficiency

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2014/0155475 A1
- Document ID: US20140155475A1
- Status: Published application
- Kind: A1
- Application number: 14/170,914
- Filed: 2014-02-03
- Published: 2014-06-05
- Earliest claimed: 2012-04-02
- Assignees: Moderna Therapeutics, Inc.
- Applicants: Moderna Therapeutics, Inc.
- Inventors: Stephane Bancel; Tirtha Chakraborty; Antonin de Fougerolles; Susan Whoriskey
- Web page: https://patentgrep.com/patent/US20140155475A1

## 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 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911})
- A61K 9/145: {with organic compounds}
- A61K 9/5123: {Organic compounds, e.g. fats, sugars}
- A61K 9/5146: {obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides}
- A61K 9/5153: {Polyesters, e.g. poly(lactide-co-glycolide)}
- A61K 31/7115: Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
- A61K 38/17: from animals; from humans {(enzyme inhibitors A61K38/005)}
- A61K 38/177: {Receptors; Cell surface antigens; Cell surface determinants}
- A61K 38/1816: {Erythropoietin [EPO]}
- A61K 38/1891: {Angiogenesic factors; Angiogenin}
- A61K 38/191: {Tumor necrosis factors [TNF], e.g. lymphotoxin [LT], i.e. TNF-beta}
- A61K 38/193: {Colony stimulating factors [CSF]}
- A61K 38/212: {IFN-alpha}
- A61K 38/45: Transferases (2)
- A61K 38/4846: {Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)}
- A61K 47/543: {Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine}
- A61K 48/0033: {the non-active part being non-polymeric}
- A61K 48/0041: {the non-active part being polymeric}
- A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered}
- A61K 48/0075: {characterised by an aspect of the delivery route, e.g. oral, subcutaneous}
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00)
- A61P 3/06: Antihyperlipidemics
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 5/00: Drugs for disorders of the endocrine system
- A61P 7/00: Drugs for disorders of the blood or the extracellular fluid
- A61P 7/02: Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A61P 7/06: Antianaemics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 13/00: Drugs for disorders of the urinary system
- A61P 13/12: of the kidneys
- A61P 15/00: Drugs for genital or sexual disorders (for disorders of sex hormones A61P5/24); Contraceptives
- A61P 17/00: Drugs for dermatological disorders
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 17/04: Antipruritics
- A61P 17/06: Antipsoriatics
- A61P 17/08: Antiseborrheics
- A61P 17/14: for baldness or alopecia
- A61P 19/00: Drugs for skeletal disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 19/10: for osteoporosis
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- A61P 21/04: for myasthenia gravis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 25/02: for peripheral neuropathies
- A61P 25/14: for treating abnormal movements, e.g. chorea, dyskinesia
- A61P 25/18: Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
- A61P 25/24: Antidepressants
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 25/30: for treating abuse or dependence
- A61P 27/02: Ophthalmic agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- A61P 31/10: Antimycotics
- A61P 31/12: Antivirals
- A61P 31/14: for RNA viruses
- A61P 31/18: for HIV
- A61P 35/00: Antineoplastic agents
- A61P 35/02: specific for leukemia
- A61P 35/04: specific for metastasis
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61P 37/08: Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14)
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07H 21/02: with ribosyl as saccharide radical
- C07K 14/005: from viruses
- C07K 14/435: from animals; from humans
- C07K 14/43595: {from coelenteratae, e.g. medusae}
- C07K 14/47: from mammals
- C07K 14/4705: {stimulating, promoting or activating activity}
- C07K 14/4713: {Autoimmune diseases, e.g. Insulin-dependent diabetes mellitus, multiple sclerosis, rheumathoid arthritis, systemic lupus erythematosus; Autoantigens}
- C07K 14/4723: {Cationic antimicrobial peptides, e.g. defensins}
- C07K 14/4746: {p53}
- C07K 14/475: Growth factors; Growth regulators
- C07K 14/485: Epidermal growth factor [EGF], i.e. urogastrone
- C07K 14/495: Transforming growth factor [TGF]
- C07K 14/505: Erythropoietin [EPO]
- C07K 14/515: Angiogenesic factors; Angiogenin
- C07K 14/525: Tumour necrosis factor [TNF]
- C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF
- C07K 14/5418: {IL-7}
- C07K 14/56: IFN-alpha
- C07K 14/61: Growth hormone [GH], i.e. somatotropin
- C07K 14/62: Insulins
- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- C07K 14/745: Blood coagulation or fibrinolysis factors
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C07K 16/2863: {against receptors for growth factors, growth regulators}
- C07K 16/2887: {against CD20}
- C07K 16/40: against enzymes
- C12N 9/0069: {acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)}
- C12N 9/0091: {oxidizing metal ions (1.16)}
- C12N 9/1051: {Hexosyltransferases (2.4.1)}
- C12N 9/1241: {Nucleotidyltransferases (2.7.7)}
- C12N 9/16: acting on ester bonds (3.1)
- C12N 9/2402: {hydrolysing O- and S- glycosyl compounds (3.2.1)}
- C12N 9/2445: {Beta-glucosidase (3.2.1.21)}
- C12N 9/6435: {Plasmin (3.4.21.7), i.e. fibrinolysin}
- C12N 9/6437: {Coagulation factor VIIa (3.4.21.21)}
- C12N 9/644: {Coagulation factor IXa (3.4.21.22)}
- C12N 9/6443: {Coagulation factor XIa (3.4.21.27)}
- C12N 9/6451: {Coagulation factor XIIa (3.4.21.38)}
- C12N 9/88: Lyases (4.)
- C12N 9/93: {Ligases (6)}
- C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity}
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12N 15/85: for animal cells
- C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle
- C12P 13/04: Alpha- or beta- amino acids {(other amino acids C12P13/005)}
- C12P 21/00: Preparation of peptides or proteins (single cell protein C12N1/00)
- C12P 21/005: {Glycopeptides, glycoproteins}
- C12Y 603/02019: Ubiquitin-protein ligase (6.3.2.19), i.e. ubiquitin-conjugating enzyme
- 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)
- C12Y 113/12007: Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase
- C12Y 116/03001: Ferroxidase (1.16.3.1), i.e. ceruloplasmin
- C12Y 207/07012: UDP-glucose--hexose-1-phosphate uridylyltransferase (2.7.7.12)
- C12Y 304/21007: Plasmin (3.4.21.7), i.e. fibrinolysin
- C12Y 304/21022: Coagulation factor IXa (3.4.21.22)
- C12Y 304/21027: Coagulation factor XIa (3.4.21.27)
- C12Y 403/02001: Argininosuccinate lyase (4.3.2.1)
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- 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)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

## Claims

### Claim 1

A method of producing a polypeptide of interest in a mammalian cell or tissue comprising contacting said mammalian cell or tissue with a modified mRNA, wherein said modified mRNA encodes a member selected from the group consisting of lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1127), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1128), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1129), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1130), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1131), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1132), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1133), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1134), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1135), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1136) and lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1137), and wherein said modified mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.

### Claim 2 (depends on 1)

The method of claim 1, wherein the modified mRNA comprises the modification 1-methylpseudouridine and further comprises the modification 5-methylcytosine.

### Claim 3 (depends on 1)

The method of claim 1, wherein the modified mRNA comprises the modifications 5-methylcytosine and 2-thiouridine, wherein 25% of the cytosine residues in the modified mRNA are 5-methylcytosines and 25% of the uridine residues in the modified mRNA are 2-thiouridine.

### Claim 4 (depends on 1)

The method of claim 1, wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the modified mRNA is not derived from beta-globin.

### Claim 5

A modified mRNA encoding a polypeptide of interest, said polypeptide of interest selected from the group consisting of lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1127), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1128), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1129), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1130), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1131), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1132), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1133), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1134), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1135), lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1136) and lipase A, lysosomal acid, cholesterol esterase (SEQ ID NO: 1137), and wherein said modified mRNA comprises modifications selected from the group consisting of:
(a) 5-methylcytosine and 2-thiouridine,
(b) pseudouridine and 5-methylcytosine, and
(c) 1-methylpseudouridine.

### Claim 6 (depends on 5)

The modified mRNA of claim 5, wherein the modified mRNA comprises the modification 1-methylpseudouridine and further comprises the modification 5-methylcytosine.

### Claim 7 (depends on 5)

The modified mRNA of claim 5, wherein the modified mRNA comprises the modifications 5-methylcytosine and 2-thiouridine, wherein 25% of the cytosine residues in the modified mRNA are 5-methylcytosines and 25% of the uridine residues in the modified mRNA are 2-thiouridine.

### Claim 8 (depends on 5)

The modified mRNA of claim 5, wherein at least one of the 5′ untranslated region or the 3′ untranslated region of the modified mRNA is not derived from beta-globin.

### Claim 9

(canceled)