Engineered nucleic acids and methods of use thereof

Patent metadata

Field	Value
Publication number	US-2018271938-A1
Application number	US-201815981762-A
Country	US
Kind code	A1
Filing date	May 16, 2018
Priority date	Sep 12, 2011
Publication date	Sep 27, 2018
Grant date	—

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

1 - 23 . (canceled)

24 . 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.

25 . The lipid-based nanoparticle of claim 24 , wherein the LL-37 polypeptide comprises the sequence of SEQ ID NO: 6.

26 . The lipid-based nanoparticle of claim 24 , 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.

27 . The lipid-based nanoparticle of claim 26 , wherein the route is intravenous.

28 . The lipid-based nanoparticle of claim 24 , 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, wybuto sine, 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.

29 . The lipid-based nanoparticle of claim 28 , wherein the nucleoside modification is pseudouridine.

30 . The lipid-based nanoparticle of claim 29 , wherein 100% of uridine nucleotides in the synthetic mRNA are modified.

31 . The lipid-based nanoparticle of claim 24 , 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.

32 . A pharmaceutical composition comprising the lipid-based nanoparticle of claim 24 .

33 . A method of treating a microbial infection comprising administering to a subject having a microbial infection the pharmaceutical composition of claim 32 .

34 . The lipid-based nanoparticle of claim 24 , 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.

35 . The lipid-based nanoparticle of claim 34 , wherein the nucleoside modification is pseudouridine.

36 . The lipid-based nanoparticle of claim 24 , wherein the LL-37 polypeptide comprises the sequence of SEQ ID NO: 6.

37 . 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.

38 . The kit of claim 37 , wherein the nucleoside modification is pseudouridine.

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

40 . A lipid-based nanoparticle comprising a synthetic messenger ribonucleic acid (mRNA) encoding a RNase-7 polypeptide in an amount effective to permit production of the RNase-7 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.

41 . A method of treating a microbial infection comprising administering to a subject having a microbial infection a pharmaceutical composition comprising the lipid-based nanoparticle of claim 40 .

42 . A lipid-based nanoparticle comprising a synthetic messenger ribonucleic acid (mRNA) encoding a defensin polypeptide in an amount effective to permit production of the defensin 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.

43 . A method of treating a microbial infection comprising administering to a subject having a microbial infection a pharmaceutical composition comprising the lipid-based nanoparticle of claim 42 .

Assignees

• Modernatx Inc

Inventors

• Bancel Stephane
• Schrum Jason P
• Aristarkhov Alexander

Classifications

• A61K38/465

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

• A61K48/0075

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

• C07K14/4723

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

• C12Y301/27

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

• A61K45/06

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