# US 12,109,275 B2 · Nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal for increasing the expression of an encoded protein

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

- Number: US 12,109,275 B2
- Document ID: US12109275B2
- Status: Granted
- Kind: B2
- Application number: 16/848,991
- Filed: 2020-04-15
- Published: 2020-09-03
- Granted: 2024-10-08
- Earliest claimed: 2010-08-13
- Assignees: CureVac AG
- Applicants: CureVac SE
- Inventors: Andreas Thess; Thomas Schlake; Jochen Probst
- Web page: https://patentgrep.com/patent/US12109275B2

## 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 39/0011: {Cancer antigens}
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00)
- A61P 3/02: Nutrients, e.g. vitamins, minerals
- A61P 5/00: Drugs for disorders of the endocrine system
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 17/00: Drugs for dermatological disorders
- A61P 19/00: Drugs for skeletal disorders
- A61P 21/00: Drugs for disorders of the muscular or neuromuscular system
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/02: Ophthalmic agents
- A61P 27/16: Otologicals
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- A61P 31/10: Antimycotics
- A61P 31/12: Antivirals
- A61P 33/00: Antiparasitic agents
- A61P 35/00: Antineoplastic agents
- 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
- C07K 14/435: from animals; from humans
- C12N 15/67: General methods for enhancing the expression
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/575: {humoral response}
- 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
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present application describes a coding nucleic acid sequence, particularly a messenger RNA (mRNA), comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal and the use thereof for increasing the expression of an encoded protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine e.g. for the use in the treatment of tumours and cancer diseases, cardiovascular diseases, infectious diseases, autoimmune diseases or genetic diseases, or in gene therapy. The present invention further describes an in vitro transcription method, in vitro methods for increasing the expression of a protein using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal and an ex vivo and in vivo method.

## Claims

### Claim 1

A purified RNA molecule comprising:
a) a coding region encoding a polypeptide;
b) at least one histone stem-loop comprising the RNA sequence of SEQ ID NO: 27; and
c) a poly(A) sequence of about 25 to about 400 adenosine nucleotides.

### Claim 2 (depends on 1)

The RNA molecule of claim 1, wherein the coding region does not code for: a histone polypeptide; a reporter polypeptide selected from EGFP and Luciferase; or a marker polypeptide selected from alpha-Globin, Galactokinase and Xanthine:guanine phosphoribosyl transferase (GPT).

### Claim 3 (depends on 1)

The RNA molecule of claim 1, comprising a poly(A) sequence of about 50 to about 250 nucleotides.

### Claim 4 (depends on 1)

The RNA molecule of claim 1, comprising from 5′ to 3′: the coding region; the poly(A) sequence; and the at least one histone stem-loop.

### Claim 5 (depends on 1)

The RNA molecule of claim 1, comprising from 5′ to 3′: the coding region; the at least one histone stem-loop; and the poly(A) sequence.

### Claim 6 (depends on 5)

The RNA molecule of claim 5, wherein the poly(A) sequence is at the 3′ end of the RNA molecule.

### Claim 7 (depends on 1)

The RNA molecule of claim 1, further comprising a methylated 5′ Cap.

### Claim 8 (depends on 7)

The RNA molecule of claim 7, wherein the methylated 5′ Cap comprises m7GpppN.

### Claim 9 (depends on 1)

The RNA molecule of claim 1, further comprising a poly(C) sequence of at least 10 cytidines.

### Claim 10 (depends on 9)

The RNA molecule of claim 9, wherein the poly(C) sequence is about 10 to about 200 cytidines.

### Claim 11 (depends on 1)

The RNA molecule of claim 1, further comprising a 5′ and/or 3′ untranslated region (UTR).

### Claim 12 (depends on 11)

The RNA molecule of claim 11, wherein the coding region encodes an antigen and wherein the RNA comprises a methylated 5′ Cap.

### Claim 13 (depends on 1)

The RNA molecule of claim 1, wherein the coding region encodes an infectious disease antigen.

### Claim 14 (depends on 13)

The RNA molecule of claim 13, wherein the infectious disease antigen is a viral antigen.

### Claim 15 (depends on 14)

The RNA molecule of claim 14, wherein the viral antigen is an antigen from influenza, rabies, severe acute respiratory syndrome (SARS), or herpes virus.

### Claim 16 (depends on 1)

The RNA molecule of claim 1, wherein the coding region encodes a tumor antigen.

### Claim 17 (depends on 1)

A pharmaceutical composition comprising the RNA molecule of claim 1 and pharmaceutically acceptable carrier.

### Claim 18 (depends on 17)

The pharmaceutical composition of claim 17, wherein the RNA molecule is in complex with a cationic compound.

### Claim 19 (depends on 17)

The pharmaceutical composition of claim 17, wherein the cation compound is a cation lipid.