# US 10,501,513 B2 · Modified polynucleotides for the production of oncology-related proteins and peptides

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- Number: US 10,501,513 B2
- Document ID: US10501513B2
- Status: Granted
- Kind: B2
- Application number: 15/425,813
- Filed: 2017-02-06
- Published: 2018-01-04
- Granted: 2019-12-10
- Earliest claimed: 2012-04-02
- Assignees: ModernaTX, Inc.
- Applicants: ModernaTX, Inc.
- Inventors: Antonin De Fougerolles; Justin Guild
- Web page: https://patentgrep.com/patent/US10501513B2

## CPC

- C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF
- 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 9/0043: {Nose}
- A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911})
- A61K 9/1272: comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers (lipids as modifying agents {A61K47/543})
- A61K 9/1277: Preparation processes; Proliposomes
- A61K 9/14: Particulate form, e.g. powders, {Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles (microspheres A61K9/16; microcapsules A61K9/50; nanocapsules, nanoparticles of the matrix type A61K9/51)}
- A61K 9/145: {with organic compounds}
- A61K 9/5031: {obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)}
- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 31/7115: Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
- A61K 38/1767: {from invertebrates}
- A61K 38/1816: {Erythropoietin [EPO]}
- A61K 38/1866: {Vascular endothelial growth factor [VEGF]}
- 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 38/215: {IFN-beta}
- A61K 38/36: Blood coagulation or fibrinolysis factors
- A61K 38/363: {Fibrinogen}
- A61K 38/44: Oxidoreductases (1)
- A61K 38/4833: {Thrombin (3.4.21.5)}
- 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 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 47/10: Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
- A61K 47/60: the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
- A61K 48/0033: {the non-active part being non-polymeric}
- 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}
- C07K 14/47: from mammals
- C07K 14/475: Growth factors; Growth regulators
- C07K 14/505: Erythropoietin [EPO]
- C07K 14/525: Tumour necrosis factor [TNF]
- C07K 14/56: IFN-alpha
- C07K 14/565: IFN-beta
- C07K 14/745: Blood coagulation or fibrinolysis factors
- C07K 14/75: Fibrinogen
- C07K 16/2887: {against CD20}
- C07K 16/32: against translation products of oncogenes
- C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes}
- C12N 9/0069: {acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)}
- C12N 9/644: {Coagulation factor IXa (3.4.21.22)}
- C12N 15/85: for animal cells
- C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle
- C12Y 113/12007: Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase
- C12Y 304/21005: Thrombin (3.4.21.5)
- C12Y 304/21022: Coagulation factor IXa (3.4.21.22)
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- C12N 2840/00: Vectors comprising a special translation-regulating system
- 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)
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The invention relates to compositions including polynucleotides encoding polypeptides which have been chemically modified by replacing the uridines with 1-methyl-pseudouridine to improve one or more of the stability and/or clearance in tissues, receptor uptake and/or kinetics, cellular access by the compositions, engagement with translational machinery, mRNA half-life, translation efficiency, immune evasion, protein production capacity, secretion efficiency, accessibility to circulation, protein half-life and/or modulation of a cell's status, function, and/or activity.

## Claims

### Claim 1

A pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a cationic lipid, a non-cationic lipid, cholesterol, and a PEG lipid, wherein the plurality of lipid nanoparticles has a mean particle size of between 80 nm and 150 nm and encapsulate an mRNA comprising:
(a) an open reading frame encoding an oncology-related protein and consisting of nucleotides selected from N1-methyl-pseudouridine, cytidine, adenosine, and guanosine;
(b) a 5′-untranslated region (UTR);
(c) at least one 5′ cap structure;
(d) a 3′-UTR; and
(e) a 3′ tailing sequence of linked nucleosides.

### Claim 2 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 5′ untranslated region is heterologous to the coding region of the mRNA.

### Claim 3 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 3′ untranslated region is heterologous to the coding region of the mRNA.

### Claim 4 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 5′ untranslated region and the 3′ untranslated region are heterologous to the coding region of the mRNA.

### Claim 5 (depends on 1)

The pharmaceutical composition of claim 1, wherein the mRNA comprises at least two stop codons.

### Claim 6 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 3′-UTR comprises comprises at least one miR binding site.

### Claim 7 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 3′-UTR comprises a miR-122 binding site.

### Claim 8 (depends on 1)

The pharmaceutical composition of claim 1, wherein the 3′-tailing sequence of linked nucleosides is a poly-A tail or a polyA-G quartet.

### Claim 9 (depends on 1)

The pharmaceutical composition of claim 1, wherein the at least one 5′ terminal cap is selected from the group consisting of Cap0, Cap1, ARCA, inosine, N1-methyl-guanosine, 2′fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

### Claim 10 (depends on 1)

The pharmaceutical composition of claim 1, wherein the cationic lipid is a biodegradable cationic lipid.

### Claim 11 (depends on 10)

The pharmaceutical composition of claim 10, wherein the cationic lipid comprises at least one ester linkage.

### Claim 12 (depends on 1)

The pharmaceutical composition of claim 1, wherein the non-cationic lipid is a phospholipid.