Gene therapy for treating citrullenemia

Patent metadata

Field	Value
Publication number	US-11535866-B2
Application number	US-201816478971-A
Country	US
Kind code	B2
Filing date	Feb 1, 2018
Priority date	Feb 1, 2017
Publication date	Dec 27, 2022
Grant date	Dec 27, 2022

Abstract

Compositions and regimens useful in treating type I citrullenemia are provided. The compositions include recombinant adeno-associated virus (rAAV) with a transthyretin enhancer and promoter driving expression of a human Argininosuccinate Synthase 1 (ASS1).

First claim

The invention claimed is:

1. A recombinant adeno-associated virus (rAAV) useful as a liver-directed therapeutic for citrullinemia, said rAAV comprising an AAV capsid, and a vector genome packaged therein, said vector genome comprising:
(a) an AAV 5′ inverted terminal repeat (ITR);
(b) a promoter;
(c) a coding sequence encoding a human argininosuccinate synthase 1 (ASS1);
(d) an AAV 3′ ITR,
wherein the coding sequence of (c) is SEQ ID NO: 2.

2. The rAAV according to claim 1 , wherein the AAV capsid is an AAV8 capsid or variant thereof.

3. The rAAV according to claim 1 , wherein the promoter is a thyroxine binding globulin (TBG) promoter.

4. The rAAV according to claim 1 , wherein the promoter is an alpha 1 anti-trypsin (A1AT) promoter.

5. The rAAV according to claim 1 , wherein the promoter comprises SEQ ID NO: 21.

6. The rAAV according to claim 1 , wherein the promoter is a transthyretin promoter (TTR) promoter.

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

8. The rAAV according to claim 1 , wherein the vector genome further comprises a polyadenylation (polyA) sequence.

9. The rAAV according to claim 1 , wherein the polyA sequence is a bovine growth hormone (bGH) polyA sequence.

10. The rAAV according to claim 1 , wherein the vector genome further comprises a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE).

11. The rAAV according to claim 1 , wherein the vector genome further comprises an intron.

12. The rAAV according to claim 11 , wherein the intron is human beta globin IVS2 or SV40 intron.

13. The rAAV according to claim 1 , wherein the vector genome further comprises an enhancer.

14. The rAAV according to claim 13 , wherein the enhancer is an APB enhancer, an ABPS enhancer, an alpha mic/bik enhancer, a TTR enhancer, en34, or an ApoE enhancer.

15. The rAAV according to claim 1 , wherein the vector genome is about 3 kilobases to about 5.5 kilobases in size.

16. An aqueous suspension suitable for administration to a citrullinemia patient, said suspension comprising an aqueous suspending liquid and about 1×10 12 genome copies (GC)/mL to about 1×10 14 GC/mL of a rAAV useful as a liver-directed therapeutic for citrullinemia, said rAAV having an AAV capsid, and having packaged therein a vector genome comprising:
(a) an AAV 5′ ITR;
(b) a promoter;
(c) a coding sequence encoding an ASS1; and
(d) an AAV 3′ ITR,
wherein the coding sequence of (c) is SEQ ID NO: 2.

17. The aqueous suspension according to claim 16 , wherein the suspension is suitable for intravenous injection.

18. The aqueous suspension according to claim 16 , wherein the suspension further comprises a surfactant, preservative, and/or buffer dissolved in the aqueous suspending liquid.

19. A method of treating a patient having citrullinemia with an rAAV, the method comprising:
administering to the patient about 1×10 12 to about 1×10 14 GC/kg of the rAAV according to claim 2 in an aqueous suspension,
wherein the GC are calculated by optimized quantitative polymerase chain reaction (oqPCR) or digital droplet polymerase chain reaction (ddPCR).

20. The aqueous suspension according to claim 16 , wherein the AAV capsid is an AAV8 capsid.

Assignees

• Univ Pennsylvania

Inventors

• Wilson James M
• Sidrane Jenny Agnes
• Wang Lili

Classifications

• C12N15/86 Primary

  Viral vectors · CPC title

• C12N2830/008

  cell type or tissue specific enhancer/promoter combination · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title