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Last Updated: October 21, 2019

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Claims for Patent: 6,372,500

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Summary for Patent: 6,372,500
Title: Episomal expression cassettes for gene therapy
Abstract:The invention consists of episomal expression cassettes for expression of a transgene in gene therapy. The expression cassettes consist of regulatory elements of the human cytokeratin gene and a transgene. The invention also includes of liposomes for transfection of epithelial tissue with the cassettes in treatment of cystic fibrosis, emphysema, cancers of epithelial origin arising in the lung or other organs.
Inventor(s): Hu; Jim (East York, CA), Chow; Yu-Hua (North York, CA), O\'Brodovich; Hugh (Oakville, CA), Tsui; Lap-Chee (Etobicoke, CA)
Assignee: HSC Research and Development Limited Partnership (Toronto, CA)
Application Number:09/423,744
Patent Claims:1. An expression cassette for the episomal expression of a transgene in a targeted mammalian epithelial cell, comprising: a cytokeratin 18 5' region, a cytokeratin 18 promoter, and a cytokeratin 18 intron 1, wherein the expression cassette is capable of receiving a transgene for expression in the epithelial cell, with the proviso that the transgene is not cytokeratin 18.

2. The expression cassette of claim 1, wherein the cytokeratin 18 intron 1 comprises all or part of nucleotide nos. 2566 to 3315 of SEQ ID NO:1.

3. The expression cassette of claim 1, whwerein the cytokeratin 18 5' region and cytokeratin 18 promoter comprise all or part of nucleotide nos. 1 to 2565 of SEQ ID NO:1.

4. The expression cassette of claim 1, further comprising a translational enhancer.

5. The expression cassette of claim 4, wherein the translational enhancer comprises all or part of nucleotide nos. 3316 to 3354 of SEO ID NO:1.

6. The expression cassette of claim 1, comprising all or part of nucleotide nos. 1 to 3354 and 7949 to 12143 of SEQ ID NO:1.

7. The expression cassette of claims 1, 2, 3, 4, 5 or 6, further comprising a transgene selected from the group consisting of (i) a nucleotide sequence comprising all or part of nucleotide nos. 3355 to 7955 of SEQ ID NO:1 (ii) a nucleotide sequence having at least 80% sequence identity with nucleotide nos. 3355 to 7955 of SEQ ID NO:1 and (iii) a nucleotide sequence encoding the same polypeptide that is encoded by nucleotide nos. 3355 to 7955 of SEQ ID NO:1.

8. The expression cassette of claim 7, wherein the cassette is modified in an intron selected from the group consisting of the k18 intron I 3' splice site, a CFTR intron 3' cryptic splice site-1 and a CFTR intron 3' cryptic splice site-2 thereby reducing alternative RNA splicing and increasing the steady state level of mRNA produced from the CFTR sequence.

9. The expression cassette of claim 8, wherein the transgene comprises all or part of nucleotide numbers 3355 to 7955 of SEQ ID NO:1 and an intron selected frorm the group consisting of nucleotides 3301 to 3304, 3306 to 3308, 3310, 3315 and 3624 of SEQ ID NO:1.

10. The expression cassette of claim 2, comprising all or part of the nucleotide sequence of SEQ ID NO:1.

11. An expression cassette comprising at least 80% sequence identity with a sequence selected from the group consisting of nucleotide nos. 1 to 3354, 7949 to 12143 of SEQ ID NO:1 and SEQ ID NO:1.

12. A vector comprising the expression cassette of claim 1.

13. The expression cassette of claim 1, wherein the epithelial cell is a lung epithelial cell.

14. The expression cassette of claim 13, wherein the lung epithelial cell is an airway epithelial cell or a submucosal cell.

15. A mammalian epithelial cell comprising the expression cassette of claim 1.

16. The mammalian epithelial cell of claim 15, wherein said mammalian epithelial cell is a human cell.

17. A composition comprising an effective amount of the expression cassette of claim 7 and a pharmaceutically acceptable carrier.

18. A composition comprising the expression cassette of claim 7 and a carrier.

19. A method for expressing a gene in a target epithelial cell, comprising:

administering to the epithelial cell an amount of the expression cassette of claims 1, 2, 3, 4, 5, 6, 9, 10 or 11 so that the expression cassette is inserted in the epithelial cell; and

expressing a protein encoded by the transgene to produce the protein.

20. A method for increasing chloride channels in an epithelial cell, comprising:

administering to the epithelial cell an amount of the expression cassette of claims 1, 2, 3, 4, 5, 6, 9, 10 or 11 so that the expression cassette is inserted in the epithelial cell; and

expressing a protein encoded by the transgene to produce the protein, wherein said protein is transported to the plasma membrane and generates chloride channels in the epithelial cell.

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
Canada2205076May 14, 1997
PCT Information
PCT FiledMay 14, 1998PCT Application Number:PCT/CA98/00478
PCT Publication Date:November 19, 1998PCT Publication Number:WO98/51807

Details for Patent 6,372,500

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04   Start Trial HSC Research and Development Limited Partnership (Toronto, CA) 2017-05-14 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial HSC Research and Development Limited Partnership (Toronto, CA) 2017-05-14 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial HSC Research and Development Limited Partnership (Toronto, CA) 2017-05-14 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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