Drug Resistance Reversal Potential of Ursolic Acid Derivatives against Nalidixic Acid‐ and Multidrug‐resistant Escherichia coli

As a part of our drug discovery program, ursolic acid was chemically transformed into six semi‐synthetic derivatives, which were evaluated for their antibacterial and drug resistance reversal potential in combination with conventional antibiotic nalidixic acid against the nalidixic acid‐sensitive and nalidixic acid‐resistant strains of Escherichia coli. Although ursolic acid and its all semi‐synthetic derivatives did not show antibacterial activity of their own, but in combination, they significantly reduced the minimum inhibitory concentration of nalidixic acid up to eightfold. The 3‐O‐acetyl‐urs‐12‐en‐28‐isopropyl ester (UA‐4) and 3‐O‐acetyl‐urs‐12‐en‐28‐n‐butyl ester (UA‐5) derivatives of ursolic acid reduced the minimum inhibitory concentration of nalidixic acid by eightfold against nalidixic acid‐resistant and four and eightfold against nalidixic acid‐sensitive, respectively. The UA‐4 and UA‐5 were further evaluated for their synergy potential with another antibiotic tetracycline against the multidrug‐resistant clinical isolate of Escherichia coli‐KG4. The results showed that both these derivatives in combination with tetracycline reduced the cell viability in concentration‐dependent manner by significantly inhibiting efflux pump. This was further supported by the in silico binding affinity of UA‐4 and UA‐5 with efflux pump proteins. These ursolic acid derivatives may find their potential use as synergistic agents in the treatment of multidrug‐resistant Gram‐negative infections.

[1]  L. Piddock.  Multidrug-resistance efflux pumps ? not just for resistance , 2006, Nature Reviews Microbiology.

[2]  B. Marquez.  Bacterial efflux systems and efflux pumps inhibitors. , 2005, Biochimie.

[3]  N. Goel,et al.  A ten year analysis of multi-drug resistant blood stream infections caused by Escherichia coli & Klebsiella pneumoniae in a tertiary care hospital , 2012, The Indian journal of medical research.

[4]  J. Molnár,et al.  New methods for the identification of efflux mediated MDR bacteria, genetic assessment of regulators and efflux pump constituents, characterization of efflux systems and screening for inhibitors of efflux pumps. , 2008, Current drug targets.

[5]  F. Mozzi,et al.  Applied Microbial and Cell Physiology , 2022 .

[6]  Jonathan R Edwards,et al.  Overview of nosocomial infections caused by gram-negative bacilli. , 2005, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[7]  A. Driessen,et al.  Distribution and Physiology of ABC-Type Transporters Contributing to Multidrug Resistance in Bacteria , 2007, Microbiology and Molecular Biology Reviews.

[8]  L. Amaral,et al.  Evaluation of efflux activity of bacteria by a semi-automated fluorometric system. , 2010, Methods in molecular biology.

[9]  Xiaohua Ma,et al.  Mechanisms of drug combinations: interaction and network perspectives , 2009, Nature Reviews Drug Discovery.

[10]  Brad Spellberg,et al.  The future of antibiotics and resistance. , 2013, The New England journal of medicine.

[11]  J. Gertsch,et al.  Botanical drugs, synergy, and network pharmacology: forth and back to intelligent mixtures. , 2011, Planta medica.

[12]  J. Pagés,et al.  Mechanisms of drug efflux and strategies to combat them: challenging the efflux pump of Gram-negative bacteria. , 2009, Biochimica et biophysica acta.

[13]  I. T. Babalola,et al.  Ubiquitous Ursolic Acid: A Potential Pentacyclic Triterpene Natural Product , 2013 .

[14]  S. Kumar.  Properties of adenyl cyclase and cyclic adenosine 3',5'-monophosphate receptor protein-deficient mutants of Escherichia coli , 1976, Journal of bacteriology.

[15]  L. Amaral,et al.  Identification of efflux-mediated multi-drug resistance in bacterial clinical isolates by two simple methods. , 2010, Methods in molecular biology.

[16]  S. Gibbons.  Plants as a Source of Bacterial Resistance Modulators and Anti-Infective Agents , 2005, Phytochemistry Reviews.

[17]  N. Koyama,et al.  Cloning, sequencing and functional expression in Escherichia coli of the gene for a P-type Na(+)-ATPase of a facultatively anaerobic alkaliphile, Exiguobacterium aurantiacum. , 2005, Biochimica et biophysica acta.

[18]  D. Méhn,et al.  Utilization of membrane vesicle preparations to study drug-ABC transporter interactions. , 2008, Expert opinion on drug metabolism & toxicology.

[19]  P. de Mol,et al.  Antimicrobial Resistance and Spread of Multi Drug Resistant Escherichia coli Isolates Collected from Nine Urology Services in the Euregion Meuse-Rhine , 2012, PloS one.

[20]  Hiroshi Nikaido,et al.  Efflux-Mediated Drug Resistance in Bacteria , 2012, Drugs.

[21]  Yanmin Hu,et al.  Enhancement by novel anti-methicillin-resistant Staphylococcus aureus compound HT61 of the activity of neomycin, gentamicin, mupirocin and chlorhexidine: in vitro and in vivo studies. , 2013, The Journal of antimicrobial chemotherapy.

[22]  J. Molnár,et al.  Inducement and Reversal of Tetracycline Resistance in Escherichia coli K-12 and Expression of Proton Gradient-Dependent Multidrug Efflux Pump Genes , 2005, Antimicrobial Agents and Chemotherapy.

[23]  P. Hawkey,et al.  The changing epidemiology of resistance. , 2009, The Journal of antimicrobial chemotherapy.

[24]  Atul Gupta,et al.  QSAR and docking studies on chalcone derivatives for antitubercular activity against M. tuberculosis H37Rv , 2014 .

[25]  S. Srivastava,et al.  Antibacterial and Synergy of Clavine Alkaloid Lysergol and its Derivatives Against Nalidixic Acid‐Resistant Escherichia coli , 2013, Chemical biology & drug design.

[26]  D. Bernhard,et al.  Ursolic acid causes DNA-damage, p53-mediated, mitochondria- and caspase-dependent human endothelial cell apoptosis, and accelerates atherosclerotic plaque formation in vivo. , 2011, Atherosclerosis.

[27]  J. Pagés,et al.  Antibiotic Stress, Genetic Response and Altered Permeability of E. coli , 2007, PloS one.

[28]  Brad Spellberg,et al.  The 10 x '20 Initiative: pursuing a global commitment to develop 10 new antibacterial drugs by 2020. , 2010, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[29]  K. Lewis,et al.  Synergy in a medicinal plant: antimicrobial action of berberine potentiated by 5'-methoxyhydnocarpin, a multidrug pump inhibitor. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[30]  I. Chopra,et al.  Targeting bacterial membrane function: an underexploited mechanism for treating persistent infections , 2010, Nature Reviews Microbiology.

[31]  Debprasad Chattopadhyay,et al.  A potent sperm motility-inhibiting activity of bioflavonoids from an ethnomedicine of Onge, Alstonia macrophylla Wall ex A. DC, leaf extract. , 2005, Contraception.

[32]  H. Nikaido.  Multidrug resistance in bacteria. , 2009, Annual review of biochemistry.

[33]  M. Hassani.  The Crisis of Resistant Gram- Negative Bacterial Infections: Is there any Hope for ESKAPE? , 2014 .

[34]  Katsunori Tanaka,et al.  Cytotoxic Activity of Ursolic Acid Derivatives Obtained by Isolation and Oxidative Derivatization , 2013, Molecules.

[35]  G. Eliopoulos,et al.  Antimicrobial Activity of Quinupristin-Dalfopristin Combined with Other Antibiotics against Vancomycin-Resistant Enterococci , 2002, Antimicrobial Agents and Chemotherapy.

[36]  M. Ferraro.  Performance standards for antimicrobial susceptibility testing , 2001 .

[37]  V. Chaturvedi,et al.  Bioenhancing and antimycobacterial agents from Ammannia multiflora. , 2012, Planta medica.

[38]  A. Yamaguchi,et al.  Novel Macrolide-Specific ABC-Type Efflux Transporter inEscherichia coli , 2001, Journal of bacteriology.

[39]  L. Piddock,et al.  The Efflux Pump Inhibitor Reserpine Selects Multidrug-Resistant Streptococcus pneumoniae Strains That Overexpress the ABC Transporters PatA and PatB , 2008, Antimicrobial Agents and Chemotherapy.

[40]  Feroz Khan,et al.  QSAR, docking and ADMET studies of camptothecin derivatives as inhibitors of DNA topoisomerase‐I , 2013 .

[41]  Ashwani Kumar,et al.  Piperine, a Phytochemical Potentiator of Ciprofloxacin against Staphylococcus aureus , 2006, Antimicrobial Agents and Chemotherapy.

[42]  J. P. Sharma,et al.  Capsaicin, a novel inhibitor of the NorA efflux pump, reduces the intracellular invasion of Staphylococcus aureus. , 2012, The Journal of antimicrobial chemotherapy.

[43]  M. Argyres,et al.  Hospital-acquired infections due to gram-negative bacteria. , 2010, The New England journal of medicine.

[44]  F. Khan,et al.  4‐Hydroxy‐α‐Tetralone and its Derivative as Drug Resistance Reversal Agents in Multi Drug Resistant Escherichia coli , 2014, Chemical biology & drug design.

[45]  J. Ruiloba.  [Antimicrobial combinations]. , 1972, Gaceta medica de Mexico.

[46]  D. Livermore.  Has the era of untreatable infections arrived? , 2009, The Journal of antimicrobial chemotherapy.