Synthesis and Characterization of Triphenyl Phosphonium-Modified Triterpenoids with Never Reported Antibacterial Effects Against Clinically Relevant Gram-Positive Superbugs

Background: To meet the urgent need for novel antibacterial agents that are active also against worrying superbugs, natural pentacyclic triterpenoids, including totally inactive betulin (BET) and betulinic acid (BA), as well as ursolic acid (UA), active on Gram-positive bacteria, have been chemically modified, achieving compounds 1–7. Methods: Triterpenoid derivatives 1–7 and all synthetic intermediates were characterized by chemometric-assisted FTIR and NMR spectroscopy, as well as by other analytical techniques, which confirmed their structure and high purity. Minimum inhibitory concentration values (MICs) of 1–7, BET, BA and UA were determined by the broth dilution method, using a selection of Gram-positive and Gram-negative clinically isolated superbugs. Results: Performed experiments evidenced that compounds 4–7 had potent antibacterial effects against Gram-positive methicillin-resistant Staphylococcus aureus and S. epidermidis (MRSA and MRSE), as well as against vancomycin-resistant Enterococcus faecalis and E. faecium (VRE). The antibacterial effects of 4–7 were due to the insertion of a triphenyl phosphonium (TPP) group and were higher than those reported so far for other BET, BA and UA derivatives, especially considering the complex pattern of resistance of the isolates used here and their clinical source. Conclusions: For the first time, by inserting TPP, a real activity (MICs 2–16 µg/mL) was conferred to inactive BET and BA (MICs > 1024 and 256 µg/mL). Moreover, the antibacterial effects of UA were improved 16- and 32-fold against MRSE and MRSA (MICs = 2 vs. 32 and 64 μg/mL). Future Perspectives: Based on these very promising microbiologic results, new experiments are currently underway with the best-performing compounds 5 and 7 (MICs = 2 μg/mL) on an enlarged number of Gram-positive isolates, to confirm their MICs. Moreover, investigations about their possible antibiofilm activity, time-killing curves and cytotoxicity on eukaryotic cells will be carried out to define their pharmacological behavior and clinical potential.