Engineered nucleic acids and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10751386-B2
Application number	US-201815981762-A
Country	US
Kind code	B2
Filing date	May 16, 2018
Priority date	Sep 12, 2011
Publication date	Aug 25, 2020
Grant date	Aug 25, 2020

Abstract

Provided are compositions and methods for delivering biological moieties such as modified nucleic acids into cells to kill or reduce the growth of microorganisms. Such compositions and methods include the use of modified messenger RNAs, and are useful to treat or prevent microbial infection, or to improve a subject's heath or wellbeing.

First claim

What is claimed is:

1. A lipid-based nanoparticle comprising a synthetic messenger ribonucleic acid (mRNA) encoding a LL-37 polypeptide in an amount effective to permit production of the LL-37 polypeptide in a cell, wherein the synthetic mRNA comprises a translatable region that contains a nucleoside modification, and wherein 75-100% of uridine nucleotides in the synthetic mRNA are modified.

2. The lipid-based nanoparticle of claim 1 , wherein the LL-37 polypeptide comprises SEQ ID NO: 6.

3. The lipid-based nanoparticle of claim 1 , wherein the composition is formulated for administration via a route selected from the group consisting of: systemic, local, intravenous, topical, oral, administration via a dressing, administration via a bandage, rectal, vaginal, intramuscular, transarterial, intraperitoneal, intranasal, subcutaneous, endoscopic, transdermal, and intrathecal.

4. The lipid-based nanoparticle of claim 3 , wherein the route is intravenous.

5. The lipid-based nanoparticle of claim 1 , wherein the nucleoside modification is selected from the group consisting of pyridin-4-one ribonucleoside, 5-aza-uridine, 2-thio-5-aza-uridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine, 5-carboxymethyl-uridine, 1-carboxymethyl-pseudouridine, 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine, 1-taurinomethyl-4-thio-uridine, 5-methyl-uridine, 1-methyl-pseudouridine, 4-thio-1-methyl-pseudouridine, 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, 5-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine, N4-acetylcytidine, 5-formylcytidine, N4-methylcytidine, 5-hydroxymethylcytidine, 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, 4-methoxy-1-methyl-pseudoisocytidine, 2-aminopurine, 2, 6-diaminopurine, 7-deaza-adenine, 7-deaza-8-aza-adenine, 7-deaza-2-aminopurine, 7-deaza-8-aza-2-aminopurine, 7-deaza-2,6-diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyladenosine, N6-methyladenosine, N6-isopentenyladenosine, N6-(cis-hydroxyisopentenyl)adenosine, 2-methylthio-N6-(cis-hydroxyisopentenyl) adenosine, N6-glycinylcarbamoyladenosine, N6-threonylcarbamoyladenosine, 2-methylthio-N6-threonyl carbamoyladenosine, N6,N6-dimethyladenosine, 7-methyladenine, 2-methylthio-adenine, 2-methoxy-adenine, inosine, 1-methyl-inosine, wyosine, wybutosine, 7-deaza-guanosine, 7-deaza-8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine, 6-thio-7-methyl-guanosine, 7-methylinosine, 6-methoxy-guanosine, 1-methylguanosine, N2-methylguanosine, N2,N2-dimethylguanosine, 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, and N2,N2-dimethyl-6-thio-guanosine.

6. The lipid-based nanoparticle of claim 5 , wherein the nucleoside modification is pseudouridine.

7. The lipid-based nanoparticle of claim 6 , wherein 100% of uridine nucleotides in the synthetic mRNA are modified.

8. The lipid-based nanoparticle of claim 1 , wherein the synthetic mRNA further comprises a 5′ untranslated region contains a nucleoside modification and/or a 3′ untranslated region that contains a nucleoside modification.

9. A method of treating a microbial infection comprising administering to a subject having a microbial infection the lipid-based nanoparticle of claim 1 .

10. The lipid-based nanoparticle of claim 1 , wherein the synthetic mRNA comprises a translatable region that comprises a nucleoside modification, a 5′ untranslated region that comprises a nucleoside modification, and a 3′ untranslated region that comprises a nucleoside modification.

11. The lipid-based nanoparticle of claim 10 , wherein the nucleoside modification is pseudouridine.

12. A kit comprising:
(a) a synthetic messenger ribonucleic acid (mRNA) encoding a LL-37 polypeptide in an amount effective to permit production of the LL-37 polypeptide in a cell, wherein the synthetic mRNA comprises a translatable region that contains a nucleoside modification, and wherein 75-100% of uridine nucleotides in the synthetic mRNA are modified; and
(b) a pharmaceutically acceptable carrier packaged in a container.

13. The kit of claim 12 , wherein the nucleoside modification is pseudouridine.

14. The kit of claim 12 , wherein the LL-37 polypeptide comprises SEQ ID NO: 6.

Assignees

• Modernatx Inc

Inventors

• Bancel Stephane
• Schrum Jason P
• Aristarkhov Alexander

Classifications

• C12Y301/27

  Endoribonucleases producing 3'-phosphomonoesters (3.1.27) · CPC title

• A61K38/465

  acting on ester bonds (3.1), e.g. lipases, ribonucleases · CPC title

• A61K38/1729 Primary

  Cationic antimicrobial peptides, e.g. defensins · CPC title

• A61K48/0075

  characterised by an aspect of the delivery route, e.g. oral, subcutaneous · CPC title

• A61K45/06

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title