Gene therapy for treating hemophilia A

Patent metadata

Field	Value
Publication number	US-11779656-B2
Application number	US-201716093798-A
Country	US
Kind code	B2
Filing date	Apr 13, 2017
Priority date	Apr 15, 2016
Publication date	Oct 10, 2023
Grant date	Oct 10, 2023

Abstract

Compositions and regimens useful in treating hemophilia A are provided. The compositions include recombinant adeno-associated virus (rAAV) with a transthyretin enhancer and promoter driving expression of a human Factor VIII.

First claim

The invention claimed is:

1. A recombinant adeno-associated virus (rAAV) useful as a liver-directed therapeutic for hemophilia A, said rAAV comprising an AAV capsid, and a vector genome packaged therein, said vector genome comprising:
(a) an AAV 5′ inverted terminal repeat (ITR) sequence;
(b) a transthyretin enhancer (enTTR) comprising the nucleotide sequence as set forth in SEQ ID NO: 5;
(c) a transthyretin (TTR) promoter consisting of the nucleotide sequence as set forth in SEQ ID NO: 7;
(d) a coding sequence encoding a human Factor VIII having coagulation function,
wherein the coding sequence comprises the nucleotide sequence as set forth in SEQ ID NO: 2;
(e) a polyA sequence; and
(f) an AAV 3′ ITR sequence,
wherein the coding sequence of (d) is operably linked to expression control elements that consist of the enTTR, the TTR promoter, and the polyA sequence.

2. The rAAV according to claim 1 , wherein the human Factor VIII is a B-domain deleted factor VIII SQ which is about 1457 amino acid residues in length.

3. The rAAV according to claim 1 , wherein the AAV capsid is an hu37 capsid.

4. The rAAV according to claim 1 , wherein the AAV 5′ ITR sequence and/or the AAV 3′ ITR sequence is from AAV2.

5. The rAAV according to claim 1 , wherein the polyA sequence is about 75 bp in length.

6. The rAAV according to claim 1 , wherein the vector genome is 5 kilobases to 5.5 kilobases in size.

7. An aqueous suspension suitable for administration to a hemophilia A patient, said suspension comprising an aqueous suspending liquid and an effective amount of a recombinant adeno-associated virus (rAAV) useful as a liver-directed therapeutic for hemophilia A, said rAAV having an AAV capsid, and having packaged therein a vector genome comprising:
(a) an AAV 5′ inverted terminal repeat (ITR) sequence;
(b) a transthyretin enhancer (enTTR) comprising the nucleotide sequence as set forth in SEQ ID NO: 5;
(c) a transthyretin (TTR) promoter consisting of the nucleotide sequence as set forth in SEQ ID NO: 7;
(d) a coding sequence encoding a human Factor VIII having coagulation function,
wherein the coding sequence comprises the nucleotide sequence as set forth in SEQ ID NO: 2;
(e) a polyA sequence; and
(f) an AAV 3′ ITR sequence,
wherein the coding sequence of (d) is operably linked to expression control elements that consist of the enTTR, the TTR promoter, and the polyA sequence.

8. The aqueous suspension according to claim 7 , wherein the suspension is suitable for intravenous injection.

9. The aqueous suspension according to claim 7 , wherein the suspension further comprises a surfactant, preservative, and/or buffer dissolved in the aqueous suspending liquid.

10. The rAAV according to claim 1 , wherein the vector genome comprises nucleotides 1-5110 of the nucleotide sequence as set forth in SEQ ID NO: 13.

11. The rAAV according to claim 10 , wherein the AAV capsid is an hu37 capsid.

12. The rAAV according to claim 1 , wherein the AAV 5′ ITR sequence comprises the nucleotide sequence as set forth in SEQ ID NO: 11 and the AAV 3′ ITR sequence comprises the nucleotide sequence as set forth in SEQ ID NO: 12.

13. The rAAV according to claim 1 , wherein the polyA sequence comprises the nucleotide sequence as set forth in SEQ ID NO:10.

14. The rAAV according to claim 2 , wherein the vector genome is 5.1 kilobases in size.

15. The aqueous suspension according to claim 7 , wherein the vector genome is 5 to 5.5 kilobases in size.

16. The aqueous suspension according to claim 7 , wherein the AAV capsid is an hu37 capsid.

17. The aqueous suspension according to claim 7 , wherein the effective amount of the rAAV is about 1×10 12 GC/kg to about 1×10 14 GC/kg body weight of the patient.

Assignees

• Univ Pennsylvania

Inventors

• Wang Lili
• Wilson James M
• Sidrane Jenny Agnes

Classifications

• A61K48/0058 Primary

  Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct · CPC title

• A61K9/0019 Primary

  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 title

• A61K38/37

  Factors VIII · CPC title

• A61P7/04

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

• C12N15/86 Primary

  Viral vectors · CPC title