التفاصيل البيبلوغرافية
| العنوان: |
CHIMERIC ADENOVIRUSES FOR USE IN CANCER TREATMENT |
| Document Number: |
20090227000 |
| تاريخ النشر: |
September 10, 2009 |
| Appl. No: |
12/413748 |
| Application Filed: |
March 30, 2009 |
| مستخلص: |
The present invention relates to oncolytic adenoviruses having therapeutic applications. Recombinant chimeric adenoviruses, and methods to produce them are provided. The chimeric adenoviruses of the invention comprise nucleic acid sequences derived from adenoviral serotypes classified within the subgroups B through F and demonstrate an enhanced therapeutic index. |
| Inventors: |
HARDEN, Paul (Brentwood, CA, US); HERMISTON, Terry (Corte Madera, CA, US); KUHN, Irene (Richmond, CA, US) |
| Assignees: |
BAYER SCHERING PHARMA AG (Berlin, DE) |
| Claim: |
1. A recombinant chimeric adenovirus, or a variant or derivative thereof, having a genome comprising an E2B region wherein said E2B region comprises a nucleic acid sequence derived from a first adenoviral serotype and a nucleic acid sequence derived from a second adenoviral serotype; wherein said first and second serotypes are each selected from the adenoviral subgroups B, C, D, E, or F and are distinct from each other; and wherein said chimeric adenovirus is oncolytic and demonstrates an enhanced therapeutic index for a tumor cell. |
| Claim: |
2. The adenovirus of claim 1 further comprising regions encoding fiber, hexon, and penton proteins, wherein the nucleic acid encoding the fiber, hexon, and penton proteins of said adenovirus are from the same adenoviral serotype. |
| Claim: |
3. The adenovirus of claim 1 further comprising a modified E3 region. |
| Claim: |
4. The adenovirus of claim 1 further comprising a modified E4 region. |
| Claim: |
5. The adenovirus of claim 1, wherein said tumor cell is a colon, breast, pancreas, lung, prostate, ovarian, or hemopoietic tumor cell. |
| Claim: |
6. The adenovirus of claim 5, wherein said tumor cell is a colon tumor cell. |
| Claim: |
7. The adenovirus of claim 1, wherein the nucleotide sequence of the E2B region of said adenovirus comprises SEQ ID NO: 3, or a portion thereof. |
| Claim: |
8. The adenovirus of claim 1, wherein the nucleotide sequence of said adenovirus comprises SEQ ID NO: 1. |
| Claim: |
9. A recombinant chimeric adenovirus, or a variant or derivative thereof, having a genome comprising an E2B region wherein said E2B region comprises a nucleic acid sequence derived from a first adenoviral serotype and a nucleic acid sequence derived from a second adenoviral serotype; wherein said first and second adenoviral serotypes are each selected from the adenoviral subgroups B, C, D, E, or F and are distinct from each other; wherein said chimeric adenovirus is oncolytic and demonstrates an enhanced therapeutic index for a tumor cell; and wherein said chimeric adenovirus has been rendered replication deficient through deletion of one or more adenoviral regions encoding proteins involved in adenoviral replication selected from the group consisting of E1, E2, E3 or E4. |
| Claim: |
10. The replication deficient adenovirus of claim 9, wherein the E1 and E3 regions have been deleted. |
| Claim: |
11. The replication deficient adenovirus of claim 10, further comprising a deletion of the E4 region. |
| Claim: |
12. The adenovirus of claim 1 or claim 9, further comprising a heterologous gene, wherein said heterologous gene is expressed within a cell infected with said adenovirus. |
| Claim: |
13. The adenovirus of claim 12, wherein said heterologous gene is thymidine kinase. |
| Claim: |
14. The adenovirus of claim 12, wherein said heterologous gene encodes a therapeutic protein selected from the group consisting of cytokines and chemokines, antibodies, pro-drug converting enzymes, and immunoregulatory proteins. |
| Claim: |
15. A method of inhibiting growth of a cancer cell, comprising infecting said cancer cell with the adenovirus of claim 1. |
| Claim: |
16. The method of claim 15, wherein said cancer cell is a colon cancer cell. |
| Claim: |
17. The method of claim 16, wherein the nucleotide sequence of said adenovirus comprises SEQ ID NO: 1. |
| Claim: |
18. A method of delivering a therapeutic protein to a cell, comprising infecting the cell with the adenovirus of claim 14. |
| Claim: |
19. A method for isolating the adenovirus of claim 1, wherein said method comprises a) pooling of adenoviral serotypes representing adenoviral subgroups B-F, thereby creating an adenoviral mixture; b) passaging the pooled adenoviral mixture from step (a) on an actively growing culture of tumor cells at a particle per cell ratio high enough to encourage recombination between serotypes, but not so high as to produce premature cell death; c) harvesting the supernatant from step (b); d) infecting a quiescent culture of tumor cells with the supernatant harvested in step (c); e) harvesting the cell culture supernatant from step (d) prior to any sign of CPE; f) infecting a quiescent culture of tumor cells with the supernatant harvested in step (e); and g) isolating the virus of claim 1 from the supernatant harvested in step (f) by plaque purification. |
| Claim: |
20. The method of claim 19, wherein step (b) is performed twice before harvesting the supernatant in step (c). |
| Claim: |
21. The method of claim 19, wherein steps (e) and (f) are repeated up to 20 times prior to step (g). |
| Claim: |
22. The method of claim 19, wherein a second round of plaque purification is performed following step (g). |
| Claim: |
23. The method of claim 19, wherein the tumor cell is a colon, breast, pancreas, lung, prostate, ovarian, or hemopoietic tumor cell. |
| Claim: |
24. A recombinant chimeric adenovirus, or a variant or derivative thereof, having a genome comprising an E2B region wherein said E2B region comprises a nucleic acid sequence derived from a first adenoviral serotype and a nucleic acid sequence derived from a second adenoviral serotype; wherein said first and second adenoviral serotypes are each selected from the adenoviral subgroups B, C, D, E, or F and are distinct from each other; and wherein said chimeric adenovirus is oncolytic and demonstrates an enhanced therapeutic index for a tumor cell; produced by the method of a) pooling of adenoviral serotypes representing adenoviral subgroups B-F, thereby creating an adenoviral mixture; b) passaging the pooled adenoviral mixture from step (a) on an actively growing culture of tumor cells at a particle per cell ratio high enough to encourage recombination between serotypes, but not so high as to produce premature cell death; c) harvesting the supernatant from step (b); d) infecting a quiescent culture of tumor cells with the supernatant harvested in step (c); e) harvesting the cell culture supernatant from step (d) prior to any sign of CPE; f) infecting a quiescent culture of tumor cells with the supernatant harvested in step (e); and g) isolating said chimeric adenovirus from the supernatant harvested in step (f) by plaque purification. |
| Claim: |
25. The method of claim 24, wherein step (b) is performed twice before harvesting of the supernatant in step (c). |
| Claim: |
26. The method of claim 24, wherein steps (e) and (f) are repeated up to 20 times prior to step (g). |
| Claim: |
27. The method of claim 24, wherein a second round of plaque purification is performed following step (g). |
| Current U.S. Class: |
435/239 |
| Current International Class: |
12; 12 |
| رقم الانضمام: |
edspap.20090227000 |
| قاعدة البيانات: |
USPTO Patent Applications |
