Nucleic acid products and methods of administration thereof

• US 10,576,167 B2
• Filed: 08/17/2017
• Issued: 03/03/2020
• Est. Priority Date: 08/17/2016
• Status: Active Grant

• Current Assignee: Factor Bioscience Inc
• Original Assignee: Factor Bioscience Inc
• Inventors: Angel, Matthew, Rohde, Christopher
• Primary Examiner(s): Epps-Smith, Janet L
• Application Number:
  US15/748,132
• Publication Number:
  US 20190008985A1
• Time in Patent Office: 929 Days
• Field of Search: 424 9321, 435455
• 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 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61K 48/0058 Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
• 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 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
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/003 Peptide-nucleic acids (PNAs)
• CPC Class Codes: C12N 2310/20 involving clustered regularly interspaced short palindromic repeats [CRISPRs]
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Y 301/21 Endodeoxyribonucleases producing 5'-phosphomonoesters (3.1.21)

Abstract

The present invention relates in part to nucleic acids, including nucleic acids encoding proteins, therapeutics and cosmetics comprising nucleic acids, methods for delivering nucleic acids to cells, tissues, organs, and patients, methods for inducing cells to express proteins using nucleic acids, methods, kits and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, therapeutics, and cosmetics produced using these methods, kits, and devices.

Claims

1. A method for treating alpha-1 antitrypsin (A1AT) deficiency comprising administering to a subject in need thereof an effective amount of

(a) a synthetic RNA encoding a gene-editing protein capable of creating a double strand break in A1AT_A (SEQ ID NO:

584) and/or

(b) a synthetic RNA encoding a gene-editing protein capable of creating a double strand break in A1AT_B (SEQ ID NO;

     585),

wherein the synthetic RNA comprises one or more non-canonical nucleotides that avoid substantial cellular toxicity, and

wherein the gene-editing protein comprises;

(i) a DNA-binding domain comprising a plurality of repeat sequences and at least one of the repeat sequences comprises the amino acid sequence;

LTPvQVVAIAwxyzGHGG (SEQ ID NO;

     629) and is between 36 and 39 amino acids long, wherein;

“

v”

is Q, D or E,

“

w”

is S or N,

“

x”

is H, N, or I,

“

y”

is D, A, I, N, G, H, K, S, or null, and

“

z”

is GGKQALETVQRLLPVLCQD (SEQ ID NO;

     630) or GGKQALETVQRLLPVLCQA (SEQ ID NO;

     631); and

(ii) a nuclease domain comprising a catalytic domain of a nuclease.

2. The method of claim 1, wherein the treatment comprises administering to a subject in need thereof an effective amount of

(a) the synthetic RNA encoding the gene-editing protein capable of creating a double strand break in A1AT_A (SEQ ID NO:

584) and

(b) the synthetic RNA encoding the gene-editing protein capable of creating a double strand break in A1AT_B (SEQ ID NO;

     585).

3. The method of claim 2, wherein the administration is directed to the liver.

4. The method of claim 1, wherein the administration is directed to the liver.

5. The method of claim 1, wherein the nuclease domain is capable of forming a dimer with another nuclease domain.

6. A method for treating alpha-1 antitrypsin (A1AT) deficiency comprising administering an effective amount of a synthetic RNA encoding a gene-editing protein capable of creating a double strand break in A1AT to a subject, wherein the synthetic RNA comprises one or more non-canonical nucleotides that avoid substantial cellular toxicity, and

wherein the gene-editing protein comprises:

(i) a DNA-binding domain comprising a plurality of repeat sequences and at least one of the repeat sequences comprises the amino acid sequence;

LTPvQVVAIAwxyzGHGG (SEQ ID NO;

     629) and is between 36 and 39 amino acids long, wherein;

“

v”

is Q, D or E,

“

w”

is S or N,

“

x”

is H, N, or I,

“

y”

is D, A, I, N, G , K, S, or null, and

“

z”

is GGKQALETVQRLLPVLCQD (SEQ ID NO;

     630) or GGKQALETVQRLLPVLCQA (SEQ ID NO;

     631); and

(ii) a nuclease domain comprising a catalytic domain of a nuclease.

7. The method of claim 6, wherein the administration is directed to the liver.

8. The method of claim 7, wherein the administration is intraportal injection.

9. The method of claim 6, wherein the administration is intraportal injection.

10. The method of claim 6, wherein the nuclease domain is capable of forming a dimer with another nuclease domain.

11. A method for reversing an alpha-1 antitrypsin (A1AT) deficiency in a cell comprising

(a) obtaining a cell comprising a defective A1AT gene;

and

(b) contacting the cell with a synthetic RNA encoding a gene-editing capable of creating a double strand break in A1AT, wherein the synthetic RNA comprises one or more non-canonical nucleotides that avoid substantial cellular toxicity, and

wherein the gene-editing protein comprises;

(i) a DNA-binding domain comprising a plurality of repeat sequences and at least one of the repeat sequences comprises the amino acid sequence;

LTPvQVVAIAwxyzGHGG (SEQ ID NO;

     629) and is between 36 and 39 amino acids long, wherein;

“

v”

is Q, D or E,

“

w”

is S or N,

“

x”

is H, N, or I,

“

y”

is D, A, I, N, G , K, S, or null, and

“

z”

is GGKQALETVQRLLPVLCQD (SEQ ID NO;

     630) or GGKQALETVQRLLPVLCQA (SEQ ID NO;

     631); and

(ii) a nuclease domain comprising a catalytic domain of nuclease.

12. The method of claim 11, further comprising steps of

(c) obtaining the cell contacted in step (b);

and

(d) administering the cell to a subject in need thereof.

13. The method of claim 12, wherein the administration is directed to the liver.

14. The method of claim 11, wherein the administration is directed to the liver.

15. A method for reversing an alpha-1 antitrypsin (A1AT) deficiency in a cell comprising

(a) obtaining a cell comprising a defective A1AT gene;

and

(b) contacting the cell with one or both of a synthetic RNA encoding a gene-editing protein capable of creating a double strand break in A1AT_A (SEQ ID NO;

     584) and a synthetic RNA encoding a gene-editing protein capable of creating a double strand break in A1AT_B (SEQ ID NO;

     585),

wherein the synthetic RNA comprises one or more non-canonical nucleotides that avoid substantial cellular toxicity, and

wherein the gene-editing protein comprises;

(i) a DNA-binding domain comprising a plurality of repeat sequences and at least one of the repeat sequences comprises the amino acid sequence;

LTPvQVVAIAwxyzGHGG (SEQ ID NO;

     629) and is between 36 and 39 amino acids long, wherein;

“

v”

is Q, D or E,

“

w”

is S or N,

“

x”

is H, N, or I,

“

y”

is D, A, I, N, G , K, S, or null, and

“

z”

is GGKQALETVQRLLPVLCQD (SEQ ID NO;

     630) or GGKQALETVQRLLPVLCQA (SEQ ID NO;

     631); and

(ii) a nuclease domain comprising a catalytic domain of nuclease.

16. The method of claim 15, further comprising steps of

(c) obtaining the cell contacted in step (b);

and

(d) administering the cell to a subject in need thereof.

17. The method of claim 16, wherein the administration is directed to the liver.

18. The method of claim 15, wherein the administration is directed to the liver.

19. The method of claim 15, wherein the nuclease domain is capable of forming a dimer with another nuclease domain.