Pactamycin analogs and methods of making thereof
This disclosure describes the molecular cloning of a pactamycin biosynthetic gene cluster from Streptomyces pactum ATCC 27456, characterization of individual genes in the gene cluster and the proteins encoded thereby as well as uses thereof. The pactamycin gene cluster is located within an 86.35 kilobases genetic locus and includes 53 open reading frames, 26 of which are considered to be the core cluster directly involved in the biosynthesis of pactamycin. The present disclosure also relates to the use of the pactamycin biosynthetic genes located within the identified gene cluster for drug design and development purposes, including the development of pactamycin analogs that are more efficacious and less toxic. Also provided are drugs and antibiotics so produced, as well as methods of their use.
1. A compound according to the formula
wherein R 1 is H or C(O)NR 4 R 5 ;
R 2 is H or lower alkyl, or hydroxyalkyl;
R 3 is H or lower alkyl; and
R 4 and R 5 independently are H or lower alkyl.
2. The compound of claim 1 , wherein R 1 is C(O)NR 4 R 5 .
3. The compound of claim 2 , wherein R 4 and R 5 are methyl.
4. The compound of claim 1 , wherein R 2 is lower alkyl or hydroxyalkyl.
5. The compound of claim 4 , wherein the lower alkyl is methyl or ethyl.
6. The compound of claim 5 , wherein the lower alkyl is methyl.
7. The compound of claim 1 , wherein R 3 is lower alkyl.
8. The compound of claim 7 , wherein the lower alkyl is methyl.
9. The compound of claim 1 , wherein R 1 is C(O)NR 4 R 5 , and R 2 , R 3 , R 4 , and R 5 are methyl.
10. A method of making the compound of claim 1 , comprising:
transforming a host cell with one or more expression vectors comprising an isolated nucleic acid molecule encoding a protein consisting of SEQ ID Nos.: 1 to 53; and
culturing the host cell in a culture medium to produce the compound of claim 1 .
11. The method of claim 10 , wherein R 1 is C(O)NR 4 R 5 .
12. The method of claim 11 , wherein R 4 and R 5 are methyl.
13. The method of claim 10 , wherein R 2 is lower alkyl or hydroxyalkyl.
14. The method of claim 13 , wherein the lower alkyl is methyl or ethyl.
15. The method of claim 14 , wherein the lower alkyl is methyl.
16. The method of claim 10 , wherein R 3 is lower alkyl.
17. The method of claim 16 , wherein the lower alkyl is methyl.
18. The method of claim 10 , wherein R 1 is C(O)NR 4 R 5 , and R 2 , R 3 , R 4 , and R 5 are methyl.