Phosphonic acid compounds and screening method
The present invention relates to pharmaceutical compositions comprising a phosphonic acid compound, such as a natural product. Such compounds can include, such as an antibiotic or compound with other activity, derived from an Actinobacteria strain having a gene encoding pepM (phosphoenolpyruvate phosphomutase) or a pepM-dependent biosynthetic pathway. The present invention also relates to methods for treating or preventing or reducing the risk of a bacterial infection by administering a therapeutically effective or prophylactically effective amount of a phosphonic acid antibiotic, or a pharmaceutical composition containing such an antibiotic, to a patient or subject in need thereof. The present invention further relates to methods for isolating, purfying, and identifying such phosphonic acid compounds from Actinobacteria strains.
1. A method for manufacturing a medicament for treating a bacterial infection in a subject in need thereof, said medicament comprising a phosphonic acid antibiotic, said antibiotic isolated from a bacterial strain of Actinobacteria having a gene encoding pepM or a pepM-dependent biosynthetic pathway, said bacterial strain selected via the following steps:
a) reviving a bacterial strain from a library of lyophilized vials or slants,
b) isolating the genomic DNA from each strain,
c) screening the genomic DNA by PCR using four pairs of degenerate primers designed to amplify a 406-bp conserved fragment within bacterial pepM genes (forward: pepMF, CGCCGGCGTCTGCNTNGARGAYAA; reverse: pepMR, GGCGCGCATCATGTGRTTNGCVYA; pepMX, GCGCGCATCATGTGGTTNGCCCADAT; pepMW, GCGCGCATCATGTGGTTNGCRTADAT; pepMZ, GCGCGCATCATGTTGTTNCCCCADAT),
d) screening the resulting PCR products by electrophoresis to retrieve pepM sequences, and
e) selecting those strains with the pepM sequences.
2. A method according to claim 1 wherein the phosphonic acid antibiotic is selected from the group consisting of
and pharmaceutically acceptable salts, esters, and prodrugs thereof, and mixtures thereof, in the manufacture of a medicament for treating a bacterial infection in a subject in need thereof.
3. A method according to claim 1 comprising the further step of f) isolating the phosphonic acid antibioitc from the selected bacterial strains.