Modified polynucleotides for treating protein deficiency

• US 9,061,059 B2
• Filed: 02/03/2014
• Issued: 06/23/2015
• Est. Priority Date: 04/02/2012
• Status: Active Grant

• Current Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Original Assignee: Moderna Therapeutics, Inc. (Moderna, Inc.)
• Inventors: Chakraborty, Tirtha, de Fougerolles, Antonin, Weiss, Ron
• Primary Examiner(s): Ketter, Jim
• Assistant Examiner(s): GALISTEO GONZALEZ, ANTONIO
• Application Number:
  US14/171,226
• Publication Number:
  US 20140249208A1
• Time in Patent Office: 505 Days
• Field of Search: 435/69.1, 536/23.5
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7115 Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/177 Receptors; Cell surface antigens; Cell surface determinants
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• CPC Class Codes: A61K 38/193 Colony stimulating factors [CSF]
• CPC Class Codes: A61K 38/212 IFN-alpha
• CPC Class Codes: A61K 38/45 Transferases (2)
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• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: 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
• CPC Class Codes: A61K 48/0075 characterised by an aspect of the delivery route, e.g. oral, subcutaneous
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
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• CPC Class Codes: C07H 21/02 : with ribosyl as saccharide radical
• CPC Class Codes: C07K 14/005 : from viruses
• CPC Class Codes: C07K 14/435 : from animals; from humans
• CPC Class Codes: C07K 14/43595 : from coelenteratae, e.g. medusae
• CPC Class Codes: C07K 14/47 : from mammals
• CPC Class Codes: C07K 14/4705 : stimulating, promoting or activating activity
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• CPC Class Codes: C12N 15/85 : for animal cells
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Abstract

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

Claims

1. A method of producing a polypeptide of interest in a mammalian cell or tissue comprising contacting said mammalian cell or tissue with an mRNA having a 3′

untranslated region and a 5′

untranslated region, wherein said mRNA encodes heparan-alpha-glucosaminide N-acetyltransferase (SEQ ID NO;

     48935), and wherein said mRNA comprises a coding region, said coding region having at least 85% identity to SEQ ID NO;

84333.

2. The method of claim 1, wherein at least one of the 5′

untranslated region or the 3′

untranslated region of the mRNA is not derived from beta-globin.

3. The method of claim 1, wherein the mRNA comprises modifications selected from the group consisting of:

(a) 5-methylcytosine and 2-thiouridine,

(b) pseudouridine and 5-methylcytosine, and

(c) 1-methylpseudouridine.

4. The method of claim 3, wherein the mRNA comprises the modification 5-methylcytosine and 2-thiouridine.

5. The method of claim 4, wherein 25% of the cytosine residues in the mRNA are 5-methylcytosines and 25% of the uridine residues in the mRNA are 2-thiouridine.

6. The method of claim 3, wherein the mRNA comprises the modification pseudouridine and 5-methylcytosine.

7. The method of claim 3, wherein the mRNA comprises the modification 1-methylpseudouridine.

8. The method of claim 7, wherein the mRNA further comprises the modification 5-methylcytosine.

9. An mRNA encoding heparan-alpha-glucosaminide N-acetyltransferase (SEQ ID NO:

48935), said mRNA having a 3′

untranslated region and a 5′

untranslated region and wherein said mRNA comprises a coding region, said coding region having at least 85% identity to SEQ ID NO;

84333.

10. The mRNA of claim 9, wherein at least one of the 5′

untranslated region or the 3′

untranslated region of the mRNA is not derived from beta-globin.

11. The mRNA of claim 9, wherein the mRNA comprises modifications selected from the group consisting of:

(a) 5-methylcytosine and 2-thiouridine,

(b) pseudouridine and 5-methylcytosine, and

(c) 1-methylpseudouridine.

12. The mRNA of claim 11, wherein the mRNA comprises the modification 5-methylcytosine and 2-thiouridine.

13. The mRNA of claim 12, wherein 25% of the cytosine residues in the mRNA are 5-methylcytosines and 25% of the uridine residues in the mRNA are 2-thiouridine.

14. The mRNA of claim 11, wherein the mRNA comprises the modification pseudouridine and 5-methylcytosine.

15. The mRNA of claim 11, wherein the mRNA comprises the modification 1-methylpseudouridine.

16. The mRNA of claim 15, wherein the mRNA further comprises the modification 5-methylcytosine.