In vivo production of proteins

Patent metadata

Field	Value
Publication number	US-9572896-B2
Application number	US-201514824247-A
Country	US
Kind code	B2
Filing date	Aug 12, 2015
Priority date	Apr 2, 2012
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

First claim

We claim:

1. A method of producing a polypeptide of interest in vivo comprising contacting a mammalian cell, tissue or organism with at least one isolated mRNA encoding the polypeptide of interest; wherein the polypeptide of interest is apolipoprotein A-I (APOA1), and wherein the isolated mRNA encoding the polypeptide of interest has at least 80% identity to SEQ ID NO: 212.

2. The method of claim 1 , wherein the isolated mRNA comprises a 3′ tailing sequence of linked nucleosides of approximately 140 nucleotides.

3. The method of claim 1 , wherein the isolated mRNA comprises a 3′ tailing sequence of linked nucleosides of approximately 160 nucleotides.

4. The method of claim 1 , wherein the isolated mRNA comprises a 5′ terminal cap of Cap1.

5. The method of claim 1 , wherein the isolated mRNA comprises at least one chemically modified nucleoside.

6. The method of claim 5 , wherein the at least one chemically modified nucleoside 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-methylpseudouridine, 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.

7. The method of claim 1 , wherein the isolated mRNA is formulated.

8. The method of claim 7 , wherein the formulation is a lipoplex formulation.

9. The method of claim 7 , wherein the formulation comprises a lipid and wherein the lipid is selected from DLin-DMA, DLin-K-DMA, DLin-KC2-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DODMA, DSDMA, DLenDMA, reLNPs, PLGA and PEGylated lipids and mixtures thereof.

10. The method of claim 7 , wherein the isolated mRNA is administered at a total daily dose of between 1 ug and 150 ug.

11. The method of claim 1 wherein the isolated mRNA is administered in two or more equal or unequal split doses.

Assignees

• Moderna Therapeutics Inc
• Modernatx Inc

Inventors

• Bancel Stephane
• Chakraborty Tirtha
• De Fougerolles Antonin
• Elbashir Sayda M
• John Matthias
• Roy Atanu
• Whoriskey Susan
• Wood Kristy M
• Hatala Paul
• Schrum Jason P
• Ejebe Kenechi
• Ellsworth Jeff Lynn
• Guild Justin

Classifications

• A61K47/543

  Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine · CPC title

• C12N15/85

  for animal cells · CPC title

• A61K38/17

  from animals; from humans {(enzyme inhibitors A61K38/005)} · CPC title

• C12P21/005

  Glycopeptides, glycoproteins · CPC title

• C12N9/93

  Ligases (6) · CPC title