# US 12,760,073 B2 · Compositions and methods of ribonucleic acid respiratory syncytial virus (RSV) vaccines

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

- Number: US 12,760,073 B2
- Document ID: US12760073B2
- Status: Granted
- Kind: B2
- Application number: 18/781,757
- Filed: 2024-07-23
- Published: 2024-11-07
- Granted: 2026-10-13
- Earliest claimed: 2021-11-19
- Assignees: RNAimmune, Inc.
- Applicants: RNAimmune, Inc.
- Inventors: David Brown; Neeti Ananthaswamy; Renxiang Chen; Dong Shen
- Web page: https://patentgrep.com/patent/US12760073B2

## CPC

- C07K 14/005: from viruses
- A61K 39/12: Viral antigens
- A61P 31/14: for RNA viruses
- A61P 37/04: Immunostimulants
- C07K 14/135: Respiratory syncytial virus
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- G05B 23/0283: {Predictive maintenance, e.g. involving the monitoring of a system and, based on the monitoring results, taking decisions on the maintenance schedule of the monitored system; Estimating remaining useful life [RUL] (preventive maintenance, i.e. planning maintenance according to the available resources without monitoring the system G06Q10/06)}
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K 2039/51: {comprising whole cells, viruses or DNA/RNA}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/55555: {Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers}
- C07K 2319/00: Fusion polypeptide
- C07K 2319/02: containing a signal sequence
- C07K 2319/03: containing a transmembrane segment
- C12N 2760/18522: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/18534: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/18541: Use of virus, viral particle or viral elements as a vector
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G05B: CONTROL OR REGULATING SYSTEMS IN GENERAL; FUNCTIONAL ELEMENTS OF SUCH SYSTEMS; MONITORING OR TESTING ARRANGEMENTS FOR SUCH SYSTEMS OR ELEMENTS (systems for controlling or regulating non-electric variables G05D; systems for regulating electric or magnetic variables G05F; control devices or systems insofar as characterised by mechanical features only G05G)

## Abstract

Provided herein is a ribonucleic acid (RNA) encoding a fusion glycoprotein (F) protein or an immunogenic fragment thereof of a respiratory syncytial virus (RSV) comprising at least one non-naturally occurring amino acid mutation. Additionally provided are relevant polynucleotides, vectors, cells, compositions, kits, production methods and methods of use.

## Claims

### Claim 1

A respiratory syncytial virus (RSV) ribonucleic acid (RNA) encoding an RSV fusion glycoprotein (F) protein or an immunogenic fragment thereof, wherein the RSV RNA comprises SEQ ID NO: 97.

### Claim 2 (depends on 1)

The RNA of claim 1, wherein the F protein further comprises one or more non-naturally occurring amino acid mutations selected from:
a histidine (H) as the amino acid corresponding to D486 of SEQ ID NO: 87 (D486H),
a glutamine (Q) as the amino acid corresponding to E487 of SEQ ID NO: 87 (E487Q),
a tryptophan (W) as the amino acid corresponding to F488 of SEQ ID NO: 87 (F488W), or
a H as the amino acid corresponding to D489 of SEQ ID NO: 87 (D489H).

### Claim 3 (depends on 1)

The RNA of claim 1, further comprising
a 3′ UTR, optionally wherein the 3′UTR is selected from SEQ ID NOs: 18, 22, or 24;
a 5′ UTR, optionally wherein the 5′ UTR comprises SEQ ID NO: 20 or 26; or
a polyA tail, optionally wherein the polyA tail is selected from SEQ ID NOs: 27, 28, or 16.

### Claim 4 (depends on 1)

The RNA of claim 1, wherein the RNA further comprises a 3′ UTR selected from SEQ ID NOs: 18, 22, or 24, a 5′ UTR selected from SEQ ID NOs: 20 or 26, and a polyA tail selected from SEQ ID NOs: 27, 28, or 16.

### Claim 5 (depends on 1)

The RNA of claim 1, wherein the RNA is chemically modified and optionally comprises one or more of: an N1-methyl-pseudouridine residue or a pseudouridine residue, and optionally wherein at least about 50%, or at least about 70%, or about 100% of the uridine residues in the RNA are N1-methyl pseudouridine or pseudouridine.

### Claim 6 (depends on 1)

A polypeptide encoded by the RNA of claim 1, wherein the polypeptide comprises SEQ ID NO: 95.

### Claim 7 (depends on 1)

A polynucleotide encoding the RNA of claim 1, wherein the polynucleotide comprises SEQ ID NO: 96.

### Claim 8 (depends on 1)

A polynucleotide encoding the RNA of claim 1, or a polynucleotide complementary thereto, optionally wherein the polynucleotide is selected from the group of: a deoxyribonucleic acid (DNA), an RNA, a hybrid of DNA and RNA, or an analog of each thereof.

### Claim 9 (depends on 8)

A vector or isolated cell comprising the polynucleotide of claim 8.

### Claim 10 (depends on 1)

A composition comprising a carrier and the RNA of claim 1.

### Claim 11 (depends on 10)

The composition of claim 10, wherein the carrier is a pharmaceutically acceptable carrier.

### Claim 12 (depends on 1)

A method of producing the RNA of claim 1, comprising culturing a cell comprising a polynucleotide encoding the RNA of claim 1 under conditions suitable for expressing the RNA.

### Claim 13 (depends on 1)

A method of producing the RNA of claim 1, comprising contacting a DNA polynucleotide encoding the RNA of claim 1 with an RNA polymerase, adenosine triphosphate (ATP), cytidine triphosphate (CTP), guanosine-5′-triphosphate (GTP), and uridine triphosphate (UTP) or a chemically modified UTP under conditions suitable for expressing the RNA.

### Claim 14 (depends on 13)

The method of claim 13, further comprising isolating the RNA.

### Claim 15 (depends on 1)

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

### Claim 16 (depends on 15)

The composition of claim 15, wherein the pharmaceutically acceptable carrier comprises a polymeric nanoparticle that comprises a Histidine-Lysine co-polymer (HKP), optionally wherein the HKP comprises a side chain selected from SEQ ID NOs: 31-44; optionally
wherein the pharmaceutically acceptable carrier further comprises a lipid, optionally wherein the lipid comprises a cationic lipid, optionally wherein the cationic lipid is ionizable, optionally wherein the cationic lipid comprises Dlin-MC3-DMA (MC3) or dioleoyloxy-3-(trimethylammonio)propane (DOTAP) or both, or optionally wherein the lipid further comprises one or more of: a helper lipid, a cholesterol, or a PEGylated lipid, and further optionally
wherein the pharmaceutically acceptable carrier comprises a lipid nanoparticle (LNP) and optionally wherein the LNP comprises one or more of: 9-Heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino}octanoate (SM-102), 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), or an equivalent of each thereof, and optionally wherein the LNP further comprises one or more of: a helper lipid, a cholesterol, or a PEGylated lipid, and optionally wherein the helper lipid comprises one or more of: disteroylphosphatidyl choline (DSPC), Dipalmitoylphosphatidylcholine (DPPC), (2R)-3-(Hexadecanoyloxy)-2-{[(9Z)-octadec-9-enoyl]oxy}propyl 2-(trimethylazaniumyl)ethyl phosphate (POPC), or dioleoyl phosphatidylethanolamine (DOPE), and optionally wherein the cholesterol comprises a plant cholesterol or an animal cholesterol or both and further optionally wherein the PEGylated lipid comprises one or more of: PEG-c-DOMG (R-3-[(ω-methoxy-poly(ethyleneglycol)2000)carbamoyl)]-1,2-dimyristyloxypropyl-3-amine), PEG-DSG (1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol), PEG-DMG (1,2-Dimyristoyl-sn-glycerol) optionally PEG2000-DMG ((1,2-dimyristoyl-sn-glycero-3-phophoethanolamine-N-[methoxy(polyethylene glycol)-2000)], or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol), and optionally the LNP comprises SM-102, DSPC, cholesterol and PEG2000-DMG, and further optionally wherein the mass ratio of the SM-102, DSPC, cholesterol and PEG200-DMG is about 1:1:1:1 and/or wherein the molar ratio of the SM-102, DSPC, cholesterol and PEG2000-DMG is about 50:10:38.5:1.5.

### Claim 17 (depends on 1)

A method of producing the composition comprising contacting the RNA of claim 1 with an HKP, thereby the RNA and the HKP are self-assembled into nanoparticles, and optionally wherein the mass ratio of HKP and the RNA in the contacting step is about 10:1 to about 1:10, optionally 2.5:1, and optionally further comprising contacting the HKP and RNA with cationic lipid, and further optionally wherein the cationic lipid comprises Dlin-MC3-DMA (MC3) or DOTAP (dioleoyloxy-3-(trimethylammonio)propane) or both, and optionally wherein the mass ratio of the cationic lipid and the RNA in the contacting step is about 10:1 to about 1:10, optionally 1:1, and optionally wherein the mass ratio of the HKP, the mRNA and the cationic lipid in the contacting step is about 4:1:1 and optionally comprising contacting the RNA of claim 1 with a lipid, thereby the RNA and the lipid are self-assembled into lipid nanoparticles (LNPs), optionally wherein the LNPs comprise one or more of: 9-Heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino}octanoate (SM-102), 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), or an equivalent of each thereof,
the LNPs further comprise one or more of: a helper lipid, a cholesterol, or a PEGylated lipid;
the helper lipid comprises one or more of: disteroylphosphatidyl choline (DSPC), Dipalmitoylphosphatidylcholine (DPPC), (2R)-3-(Hexadecanoyloxy)-2-{[(9Z)-octadec-9-enoyl]oxy}propyl 2-(trimethylazaniumyl)ethyl phosphate (POPC), or dioleoyl phosphatidylethanolamine (DOPE);
the cholesterol comprises a plant cholesterol or an animal cholesterol or both;
the PEGylated lipid comprises one or more of: PEG-c-DOMG (R-3-[(ω-methoxy-poly(ethyleneglycol)2000)carbamoyl)]-1,2-dimyristyloxypropyl-3-amine), PEG-DSG (1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol), PEG-DMG (1,2-Dimyristoyl-sn-glycerol) optionally PEG2000-DMG ((1,2-dimyristoyl-sn-glycero-3-phophoethanolamine-N-[methoxy(polyethylene glycol)-2000)], or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol);
the LNPs comprise SM-102, DSPC, cholesterol and PEG2000-DMG;
the mass ratio of the SM-102, DSPC, cholesterol and PEG200-DMG is about 1:1:1:1 and/or wherein the molar ratio of the SM-102, DSPC, cholesterol and PEG2000-DMG is about 50:10:38.5:1.5
the contacting step is performed in a microfluidic mixer, optionally selected from a slit interdigital micromixer, or a staggered herringbone micromixer (SHM).

### Claim 18 (depends on 1)

In a subject in need thereof, a method of one or more of:
(a) preventing symptomatic RSV infection,
(b) inducing an immune response to RSV,
(c) treating a subject infected with RSV, or
(d) reducing a RSV viral load,
comprising administering to the subject the RNA of claim 1, and optionally wherein the subject is a human patient, selected from an infant, a pediatric patient, or a pregnant human or an adult 60 years old or older and optionally further comprising treating the subject with an additional therapeutic agent or an additional prophylactic agent, optionally selected from one or more of an influenza virus, a corona virus, an Ebola virus, a Human Immunodeficiency Virus (HIV), or a COVID-19, and optionally wherein the additional therapeutic agent comprises a RNA polynucleotide from another infectious virus, optionally selected from one or more of an influenza virus vaccine, a corona virus vaccine, an Ebola virus vaccine, a Human Immunodeficiency Virus (HIV) vaccine, or a COVID-19 vaccine and further optionally wherein the subject does not have, or exhibit symptoms of a RSV infection when administrated with the RNA or the composition.