# US 11,896,652 B2 · Modified factor IX, and compositions, methods and uses for gene transfer to cells, organs, and tissues

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,896,652 B2
- Document ID: US11896652B2
- Status: Granted
- Kind: B2
- Application number: 18/050,982
- Filed: 2022-10-28
- Published: 2023-05-11
- Granted: 2024-02-13
- Earliest claimed: 2015-06-23
- Assignees: The Children's Hospital of Philadelphia
- Applicants: The Children's Hospital of Philadelphia
- Inventors: Katherine A. High; Xavier Anguela
- Web page: https://patentgrep.com/patent/US11896652B2

## CPC

- A61K 38/4846: {Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)}
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- C12N 9/644: {Coagulation factor IXa (3.4.21.22)}
- C12N 15/86: Viral vectors
- C12Y 304/21022: Coagulation factor IXa (3.4.21.22)
- C12N 2750/14143: viral genome or elements thereof as genetic vector
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The invention relates to modified Factor IX coding sequence, expression cassette, vectors such as viral (e.g., lenti- or adeno-associated viral) vectors, and gene transfer methods and uses. In particular, to target Factor IX nucleic acid to cells, tissues or organs for expression (transcription) of Factor IX.

## Claims

### Claim 1

A method of treating a human subject with hemophilia B comprising: administering to said subject a therapeutically effective amount of a recombinant adeno-associated virus (rAAV) vector comprising a vector genome encapsidated by an AAV5 capsid,
wherein said vector genome comprises at least two AAV inverted terminal repeats (ITR), an expression control element conferring liver tissue-specific expression operably linked with a nucleic acid sequence encoding human Factor IX (FIX) protein, and a polyadenylation signal sequence, wherein said nucleic acid sequence encoding human FIX protein is at least 70% identical to SEQ ID NO:10, has a reduced number of CpG di-nucleotides compared to wild-type nucleic acid sequence encoding human FIX protein and encodes the same human FIX protein encoded by SEQ ID NO:10, and wherein said therapeutically effective amount of said rAAV vector is a dose ranging from 1×1013 to 1×1014 vector genomes per kilogram (vg/kg) human subject body weight.

### Claim 2 (depends on 1)

The method of claim 1, wherein said nucleic acid sequence encoding human FIX protein is at least 80% identical to SEQ ID NO:10.

### Claim 3 (depends on 2)

The method of claim 2, wherein said nucleic acid sequence encoding human FIX protein is at least 85% identical to SEQ ID NO:10.

### Claim 4 (depends on 1)

The method of claim 1, wherein said vector genome further comprises an intron.

### Claim 5 (depends on 4)

The method of claim 4, wherein said intron is positioned within said nucleic acid sequence encoding human FIX protein.

### Claim 6 (depends on 4)

The method of claim 4, wherein said intron is positioned between said promoter and said nucleic acid sequence encoding human FIX protein.

### Claim 7 (depends on 1)

The method of claim 1, wherein said vector genome is linear single-stranded DNA.

### Claim 8 (depends on 1)

The method of claim 1, wherein an AAV ITR is positioned at each end of the vector genome, and said expression control element comprises an enhancer and a promoter.

### Claim 9 (depends on 8)

The method of claim 8, wherein said AAV ITRs are AAV2 ITRs, said enhancer is a human apolipoprotein HCR enhancer, and said promoter is a human alpha-1-antitrypsin gene promoter.

### Claim 10 (depends on 1)

The method of claim 1, wherein said therapeutically effective amount of said rAAV vector is a dose selected from the group consisting of 1×1013, 2×1013, 3×1013, 4×1013, 5×1013, 6×1013 7×1013, 8×1013, 9×1013 and 1×1014 vg/kg human subject body weight.

### Claim 11 (depends on 10)

The method of claim 10, wherein said therapeutically effective amount of said rAAV vector is a dose of 2×1013 vg/kg human subject body weight.

### Claim 12 (depends on 11)

The method of claim 11, wherein the disease phenotype of said hemophilia B is severe or moderate hemophilia B and said treatment is effective to convert the disease phenotype to mild hemophilia B.

### Claim 13 (depends on 1)

The method of claim 1, wherein said treatment does not cause circulating liver enzyme levels to exceed 100% of the upper limit of normal value.

### Claim 14 (depends on 1)

The method of claim 1, wherein the disease phenotype of said hemophilia B is severe or moderate hemophilia B and said treatment is effective to convert the disease phenotype to mild hemophilia B.

### Claim 15 (depends on 14)

The method of claim 14, wherein said treatment is effective to produce greater than 5% of normal FIX activity.

### Claim 16 (depends on 14)

The method of claim 14, wherein said treatment is effective to produce greater than 30% of normal FIX activity.

### Claim 17 (depends on 14)

The method of claim 14, wherein said treatment is effective to produce 6% to 40% of normal FIX activity.

### Claim 18 (depends on 14)

The method of claim 14, wherein said treatment is effective for a sustained period of at least 12 months without immunosuppression.

### Claim 19 (depends on 1)

The method of claim 1, wherein the disease phenotype of said hemophilia B is severe or moderate hemophilia B and said treatment is effective to convert the disease phenotype to a hemophilia B disease-free state.

### Claim 20 (depends on 19)

The method of claim 19, wherein said treatment is effective to produce about 50% to 150% of normal FIX activity.

### Claim 21 (depends on 19)

The method of claim 19, wherein said treatment is effective for a sustained period of at least 12 months without immunosuppression.

### Claim 22

A method of treating a human subject with hemophilia B comprising:
administering to said human subject a therapeutically effective amount of a recombinant adeno-associated virus (rAAV) vector comprising a vector genome encapsidated by an AAV5 capsid,
wherein said vector genome comprises at least two AAV inverted terminal repeats (ITR), an expression control element conferring liver tissue-specific expression operably linked with a nucleic acid sequence encoding human Factor IX (FIX) protein, and a polyadenylation signal sequence, wherein said nucleic acid sequence encoding human FIX protein is at least 70% identical to SEQ ID NO:10 and encodes the same human FIX protein encoded by SEQ ID NO:10, and
wherein said therapeutically effective amount of said rAAV vector is a dose ranging from 1×1013 to 1×1014 vector genomes per kilogram (vg/kg) human subject body weight.

### Claim 23

A method of treating a human subject with hemophilia B comprising:
administering to said human subject a therapeutically effective amount of a recombinant adeno-associated virus (rAAV) vector comprising a vector genome encapsidated by an AAV5 capsid,
wherein said vector genome comprises at least two AAV inverted terminal repeats (ITR), an expression control element conferring liver tissue-specific expression operably linked with a nucleic acid sequence encoding human Factor IX (FIX) protein, and a polyadenylation signal sequence, wherein said nucleic acid sequence encoding human FIX protein is at least 70% identical to SEQ ID NO:10 and encodes a human FIX protein variant having an amino acid sequence at least 99% identical to that encoded by SEQ ID NO:10, amino acid number 338 thereof being leucine and invariant, and
wherein said therapeutically effective amount of said rAAV vector is a dose ranging from 1×1013 to 1×1014 vector genomes per kilogram (vg/kg) human subject body weight.