Identification of 6,8-ditrifluoromethyl Halogenated Phenazine As a Potent Bacterial Biofilm-eradicating Agent

Overview

Journal Org Biomol Chem

Publisher Royal Society of Chemistry

Date 2025 Jan 22

PMID 39841058

Authors

Qiwen Gao

Hongfen Yang

Jeremy Sheiber

Priscila Cristina Bartolomeu Halicki

Ke Liu

David Blanco

Sadie Milhous

Shouguang Jin

Kyle H Rohde

Renee M Fleeman

Robert W Huigens Iii

Affiliations

Soon will be listed here.

Abstract

Bacterial biofilms are surface-attached communities consisting of non-replicating persister cells encased within an extracellular matrix of biomolecules. Unlike bacteria that have acquired resistance to antibiotics, persister cells enable biofilms to demonstrate innate tolerance toward all classes of conventional antibiotic therapies. It is estimated that 50-80% of bacterial infections are biofilm associated, which is considered the underlying cause of chronic and recurring infections. Herein, we report a modular three-step synthetic route to new halogenated phenazine (HP) analogues from diverse aniline and nitroarene building blocks. The HPs were evaluated for antibacterial and biofilm-killing properties against a panel of lab strains and multidrug-resistant clinical isolates. Several HPs demonstrated potent antibacterial (MIC ≤ 0.39 μM) and biofilm-eradicating activities (MBEC < 10 μM) with 6,8-ditrifluoromethyl-HP 15 demonstrated remarkable biofilm-killing potencies (MBEC = 0.15-1.17 μM) against Gram-positive pathogens, including methicillin-resistant clinical isolates. Confocal microscopy showed HP 15 induced significant losses in the polysaccharide matrix in MRSA biofilms. In addition, HP 15 showed increased antibacterial activities against dormant (, MIC = 1.35 μM) when compared to replicating (MIC = 3.69 μM). Overall, this new modular route has enabled rapid access to an interesting series of potent halogenated phenazine analogues to explore their unique antibacterial and biofilm-killing properties.

References

Smith P, Koehler M, Girgis H, Yan D, Chen Y, Chen Y . Optimized arylomycins are a new class of Gram-negative antibiotics. Nature. 2018; 561(7722):189-194. DOI: 10.1038/s41586-018-0483-6. View

Mitcheltree M, Pisipati A, Syroegin E, Silvestre K, Klepacki D, Mason J . A synthetic antibiotic class overcoming bacterial multidrug resistance. Nature. 2021; 599(7885):507-512. PMC: 8549432. DOI: 10.1038/s41586-021-04045-6. View

Wright P, Seiple I, Myers A . The evolving role of chemical synthesis in antibacterial drug discovery. Angew Chem Int Ed Engl. 2014; 53(34):8840-69. PMC: 4536949. DOI: 10.1002/anie.201310843. View

Liu K, Brivio M, Xiao T, Norwood 4th V, Kim Y, Jin S . Modular Synthetic Routes to Fluorine-Containing Halogenated Phenazine and Acridine Agents That Induce Rapid Iron Starvation in Methicillin-Resistant Biofilms. ACS Infect Dis. 2022; 8(2):280-295. PMC: 9004446. DOI: 10.1021/acsinfecdis.1c00402. View

Munita J, Arias C . Mechanisms of Antibiotic Resistance. Microbiol Spectr. 2016; 4(2). PMC: 4888801. DOI: 10.1128/microbiolspec.VMBF-0016-2015. View

Wright G . Molecular mechanisms of antibiotic resistance. Chem Commun (Camb). 2011; 47(14):4055-61. DOI: 10.1039/c0cc05111j. View

Sun C, Wang Q, Brubaker J, Wright P, Lerner C, Noson K . A robust platform for the synthesis of new tetracycline antibiotics. J Am Chem Soc. 2008; 130(52):17913-27. PMC: 2681267. DOI: 10.1021/ja806629e. View

Abouelhassan Y, Garrison A, Yang H, Chavez-Riveros A, Burch G, Huigens 3rd R . Recent Progress in Natural-Product-Inspired Programs Aimed To Address Antibiotic Resistance and Tolerance. J Med Chem. 2019; 62(17):7618-7642. PMC: 6742553. DOI: 10.1021/acs.jmedchem.9b00370. View

Hogan P, Chen C, Mulvihill K, Lawrence J, Moorhead E, Rickmeier J . Large-scale preparation of key building blocks for the manufacture of fully synthetic macrolide antibiotics. J Antibiot (Tokyo). 2017; 71(2):318-325. DOI: 10.1038/ja.2017.116. View

10.

Okano A, Nakayama A, Schammel A, Boger D . Total synthesis of [Ψ[C(═NH)NH]Tpg(4)]vancomycin and its (4-chlorobiphenyl)methyl derivative: impact of peripheral modifications on vancomycin analogues redesigned for dual D-Ala-D-Ala and D-Ala-D-Lac binding. J Am Chem Soc. 2014; 136(39):13522-5. PMC: 4183650. DOI: 10.1021/ja507009a. View

11.

Jennings M, Ator L, Paniak T, Minbiole K, Wuest W . Biofilm-eradicating properties of quaternary ammonium amphiphiles: simple mimics of antimicrobial peptides. Chembiochem. 2014; 15(15):2211-5. DOI: 10.1002/cbic.201402254. View

12.

Garrison A, Abouelhassan Y, Kallifidas D, Tan H, Kim Y, Jin S . An Efficient Buchwald-Hartwig/Reductive Cyclization for the Scaffold Diversification of Halogenated Phenazines: Potent Antibacterial Targeting, Biofilm Eradication, and Prodrug Exploration. J Med Chem. 2018; 61(9):3962-3983. DOI: 10.1021/acs.jmedchem.7b01903. View

13.

Lluka T, Stokes J . Antibiotic discovery in the artificial intelligence era. Ann N Y Acad Sci. 2022; 1519(1):74-93. DOI: 10.1111/nyas.14930. View

14.

Deb C, Lee C, Dubey V, Daniel J, Abomoelak B, Sirakova T . A novel in vitro multiple-stress dormancy model for Mycobacterium tuberculosis generates a lipid-loaded, drug-tolerant, dormant pathogen. PLoS One. 2009; 4(6):e6077. PMC: 2698117. DOI: 10.1371/journal.pone.0006077. View

15.

Lewis K . The Science of Antibiotic Discovery. Cell. 2020; 181(1):29-45. DOI: 10.1016/j.cell.2020.02.056. View

16.

Darby E, Trampari E, Siasat P, Gaya M, Alav I, Webber M . Molecular mechanisms of antibiotic resistance revisited. Nat Rev Microbiol. 2022; 21(5):280-295. DOI: 10.1038/s41579-022-00820-y. View

17.

Xie J, Okano A, Pierce J, James R, Stamm S, Crane C . Total synthesis of [Ψ[C(═S)NH]Tpg4]vancomycin aglycon, [Ψ[C(═NH)NH]Tpg4]vancomycin aglycon, and related key compounds: reengineering vancomycin for dual D-Ala-D-Ala and D-Ala-D-Lac binding. J Am Chem Soc. 2011; 134(2):1284-97. PMC: 3262083. DOI: 10.1021/ja209937s. View

18.

Munoz K, Hergenrother P . Facilitating Compound Entry as a Means to Discover Antibiotics for Gram-Negative Bacteria. Acc Chem Res. 2021; 54(6):1322-1333. PMC: 7969460. DOI: 10.1021/acs.accounts.0c00895. View

19.

Niu H, Gu J, Zhang Y . Bacterial persisters: molecular mechanisms and therapeutic development. Signal Transduct Target Ther. 2024; 9(1):174. PMC: 11252167. DOI: 10.1038/s41392-024-01866-5. View

20.

Richter M, Hergenrother P . The challenge of converting Gram-positive-only compounds into broad-spectrum antibiotics. Ann N Y Acad Sci. 2018; 1435(1):18-38. PMC: 6093809. DOI: 10.1111/nyas.13598. View