Optimized human clotting factor VIII gene expression cassettes and their use

Patent metadata

Field	Value
Publication number	US-10308705-B2
Application number	US-201615017015-A
Country	US
Kind code	B2
Filing date	Feb 5, 2016
Priority date	Feb 6, 2015
Publication date	Jun 4, 2019
Grant date	Jun 4, 2019

Abstract

The invention relates to synthetic liver-specific promoters and expression constructs for producing polypeptides and functional nucleic acids in the liver of a subject. The invention further relates to Factor VIII proteins containing modifications in the amino acid sequence of the Factor VIII protein, as well as nucleic acid constructs encoding the Factor VIII proteins and methods of using these compositions to treat a bleeding disorder.

First claim

What is claimed is:

1. An isolated polynucleotide comprising a synthetic liver specific promoter, wherein the promoter comprises the nucleotide sequence as set forth in SEQ ID NO: 1.

2. The polynucleotide of claim 1 , wherein the promoter is operably linked to an intron.

3. The polynucleotide of claim 2 , wherein the intron is from heavy chain variable region (V H ) 4 (V H 4).

4. The polynucleotide of claim 3 , wherein the promoter and the intron together comprise the nucleotide sequence as set forth in SEQ ID NO: 2.

5. The polynucleotide of claim 1 , wherein the promoter is operably linked to a polynucleotide of interest.

6. The polynucleotide of claim 5 , wherein the polynucleotide of interest encodes a polypeptide or a functional nucleic acid.

7. The polynucleotide of claim 6 , wherein the polynucleotide of interest encodes Factor VIII.

8. A vector comprising the polynucleotide of claim 1 .

9. An isolated transformed cell comprising the polynucleotide of claim 1 .

10. The polynucleotide of claim 6 , wherein the polynucleotide of interest encodes a clotting factor.

11. The polynucleotide of claim 7 , wherein the polynucleotide interest encodes a B domain-deleted Factor VIII.

12. The polynucleotide of claim 11 , wherein the polynucleotide of interest comprises the nucleotide sequence as set forth in SEQ ID NO: 3.

13. The polynucleotide of claim 11 , wherein the promoter and the polynucleotide of interest comprises the nucleotide sequence as set forth in SEQ ID NO: 4.

14. The polynucleotide of claim 1 , further comprising a polyadenylation site downstream of the polynucleotide of interest.

15. The vector of claim 8 , wherein the vector is a viral vector.

16. The vector of claim 14 , wherein the vector is an adeno-associated virus vector.

Assignees

• Univ North Carolina Chapel Hill

Inventors

• Xiao Xiao
• Li Juan
• Yuan Zhenhua

Classifications

• A61P7/04

  Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents · CPC title

• C07K14/755 Primary

  Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)} · CPC title

• A61K38/00

  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 title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C12N2750/14141

  Use of virus, viral particle or viral elements as a vector · CPC title