Claims for Patent: 7,049,143

Summary for Patent: 7,049,143

Title:	High efficiency retroviral vector which contains genetically engineered cellular non-coding sequence harboring splicing acceptor
Abstract:	The present invention provides a safe and highly efficient retroviral vector derived from the MLV (murine leukemia virus) for use in gene therapy, which lacks viral coding sequences but contains the genetically engineered EF I.alpha. non-coding sequence harboring splicing acceptor.
Inventor(s):	Kim; Sun-Young (Seoul, KR), Yu; Seung-Shin (Seoul, KR), Lee; Jun-Tae (Kunsan-si, KR)
Assignee:	ViroMed Co., Ltd. (KR)
Application Number:	10/129,422
Patent Claims:	1. A retroviral vector comprising a nucleotide sequence between a 5' LTR and a 3' LTR, wherein said nucleotide sequence comprises a) a MLV minimal packaging sequence comprising a splicing donor, b) a poly-purine tract, c) a multi-cloning site, and d) a heterologous non-coding sequence comprising a splicing acceptor, wherein said heterologous non-coding sequence is located upstream of said multi-cloning site, wherein said heterologous non-coding sequence is the non-coding sequence of an EF1.alpha., and with the exception that a region of the heterologous non-coding sequence downstream of said splicing acceptor comprises a mutated sequence. 2. The vector of claim 1, wherein said heterologous non-coding sequence is the sequence of the intron and exon 2 of a human EF1.alpha., wherein the intron and exon 2 are located immediately upstream of the translation initiation codon of the human EF1.alpha., with the exception that a region of said heterologous non-coding sequence downstream of said splicing acceptor comprises a mutated sequence. 3. The vector claim 1, wherein the vector further comprises one or more foreign genes. 4. The vector of claim 1, wherein said 5'LTR comprises a heterologous promoter. 5. The vector of claim 1, wherein the vector further comprises a first nucleotide sequence downstream of said multi-cloning site, wherein said first nucleotide sequence is a heterologous promoter. 6. The vector of claim 1, further comprising a nucleotide sequence encoding a selectable marker. 7. A mammalian host cell transformed with the vector of claim 1. 8. The vector of claim 1, wherein the vector is in a plasmid. 9. A cell comprising the vector of claim 1. 10. A composition comprising the vector of claim 1 and a suitable carrier. 11. The vector of claim 1, wherein the vector further comprises a first nucleotide sequence downstream of said multi-cloning site, wherein said first nucleotide sequence is an internal ribosome entry site. 12. The vector of claim 2, wherein said heterologous non-coding sequence is SEQ ID NO:1, with the exception that a region of said heterologous non-coding sequence downstream of said splicing acceptor comprises a mutated sequence. 13. The vector of claim 12, wherein said mutated sequence is at the 205th and 206th nucleotides of SEQ ID NO:1. 14. The vector of claim 13, wherein said mutated sequence is CC (cytosine-cytosine). 15. The vector of claim 3, wherein said foreign gene is a reporter gene. 16. The vector of claim 3, wherein said foreign gene is an IL-Ira gene. 17. The vector of claim 15, wherein said reporter gene is a luciferase gene. 18. The vector of claim 4, wherein said heterologous promoter is an HCMV IE promoter. 19. The vector of claim 5, wherein said heterologous promoter is an SV40 promoter. 20. The vector of claim 19, further comprising one or more foreign genes. 21. The vector of claim 6, wherein said selectable marker is a neomcyin resistance gene or a multi-drug resistance gene. 22. The mammalian host cell of claim 7, wherein said heterologous non-coding sequence is SEQ ID NO:1, with the exception that the 205th and 206th nucleotides of SEQ ID NO:1 are CC (cytosine-cytosine). 23. The vector of claim 8, wherein said heterologous non-coding sequence is SEQ ID NO:1, with the exception that the 205th and 206th nucleotides of SEQ ID NO:1 are CC (cytosine-cytosine). 24. A cell of claim 9, wherein the vector is in a plasmid. 25. The cell of claim 9, wherein the cell is a bacterial cell. 26. The cell of claim 9, wherein the cell is a packaging cell line. 27. A method of making infectious retroviral particles comprising cultivating the packaging cell line of claim 26 in a suitable medium, collecting the medium, and filtering the collected medium to obtain a cell-free viral supernatant. 28. A method of transducing mammalian cells comprising incubating the mammalian cells with the cell-free viral supernatant of claim 27. 29. The composition of claim 10, wherein said heterologous non-coding sequence is SEQ ID NO:1, with the exception that the 205th and 206th nucleotides of SEQ ID NO:1 are CC (cytosine-cytosine).

Details for Patent 7,049,143

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2020-09-08
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2020-09-08
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2020-09-08