Mycobacterium tuberculosis Shikimate Pathway Enzymes as Targets for the Rational Design of Anti-Tuberculosis Drugs

Roughly a third of the world’s population is estimated to have latent Mycobacterium tuberculosis infection, being at risk of developing active tuberculosis (TB) during their lifetime. Given the inefficacy of prophylactic measures and the increase of drug-resistant M. tuberculosis strains, there is a clear and urgent need for the development of new and more efficient chemotherapeutic agents, with selective toxicity, to be implemented on patient treatment. The component enzymes of the shikimate pathway, which is essential in mycobacteria and absent in humans, stand as attractive and potential targets for the development of new drugs to treat TB. This review gives an update on published work on the enzymes of the shikimate pathway and some insight on what can be potentially explored towards selective drug development.

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Rational Design of Biosafety Level 2-Approved, Multidrug-Resistant Strains ofMycobacterium tuberculosisthrough Nutrient Auxotrophy

mBio ◽

10.1128/mbio.00938-18 ◽

2018 ◽

Vol 9 (3) ◽

Cited By ~ 9

Author(s):

Catherine Vilchèze ◽

Jacqueline Copeland ◽

Tracy L. Keiser ◽

Torin Weisbrod ◽

Jacqueline Washington ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Rational Design ◽

Multidrug Resistant ◽

New Drugs ◽

Drug Resistant ◽

Content Type ◽

Auxotrophic Mutants ◽

Level 3 ◽

Resistant Mutants ◽

Biosafety Level

ABSTRACTMultidrug-resistant (MDR) tuberculosis, defined as tuberculosis resistant to the two first-line drugs isoniazid and rifampin, poses a serious problem for global tuberculosis control strategies. Lack of a safe and convenient model organism hampers progress in combating the spread of MDR strains ofMycobacterium tuberculosis. We reasoned that auxotrophic MDR mutants ofM. tuberculosiswould provide a safe means for studying MDRM. tuberculosiswithout the need for a biosafety level 3 (BSL3) laboratory. Two different sets of triple auxotrophic mutants ofM. tuberculosiswere generated, which were auxotrophic for the nutrients leucine, pantothenate, and arginine or for leucine, pantothenate, and methionine. These triple auxotrophic strains retained their acid-fastness, their ability to generate both a drug persistence phenotype and drug-resistant mutants, and their susceptibility to plaque-forming mycobacterial phages. MDR triple auxotrophic mutants were obtained in a two-step fashion, selecting first for solely isoniazid-resistant or rifampin-resistant mutants. Interestingly, selection for isoniazid-resistant mutants of the methionine auxotroph generated isolates with single point mutations inkatG, which encodes an isoniazid-activating enzyme, whereas similar selection using the arginine auxotroph yielded isoniazid-resistant mutants with large deletions in the chromosomal region containingkatG. TheseM. tuberculosisMDR strains were readily sterilized by second-line tuberculosis drugs and failed to kill immunocompromised mice. These strains provide attractive candidates forM. tuberculosisbiology studies and drug screening outside the BSL3 facility.IMPORTANCEElimination ofMycobacterium tuberculosis, the bacterium causing tuberculosis, requires enhanced understanding of its biology in order to identify new drugs against drug-susceptible and drug-resistantM. tuberculosisas well as uncovering novel pathways that lead toM. tuberculosisdeath. To circumvent the need for a biosafety level 3 (BSL3) laboratory when conducting research onM. tuberculosis, we have generated drug-susceptible and drug-resistant triple auxotrophic strains ofM. tuberculosissuitable for use in a BSL2 laboratory. These strains originate from a double auxotrophicM. tuberculosisstrain, H37Rv ΔpanCDΔleuCD, which was reclassified as a BSL2 strain based on its lack of lethality in immunocompromised and immunocompetent mice. A third auxotrophy (methionine or arginine) was introduced via deletion ofmetAorargB, respectively, sinceM. tuberculosisΔmetAandM. tuberculosisΔargBare unable to survive amino acid auxotrophy and infect their host. The resulting triple auxotrophicM. tuberculosisstrains retained characteristics ofM. tuberculosisrelevant for most types of investigations.

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Activity of Semi-Synthetic Mulinanes against MDR, Pre-XDR, and XDR Strains of Mycobacterium tuberculosis

Metabolites ◽

10.3390/metabo11120876 ◽

2021 ◽

Vol 11 (12) ◽

pp. 876

Author(s):

María Alejandrina Martínez-González ◽

Luis Manuel Peña-Rodríguez ◽

Andrés Humberto Uc-Cachón ◽

Jorge Bórquez ◽

Mario J. Simirgiotis ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Infectious Agent ◽

Vero Cells ◽

Bactericidal Effect ◽

Multidrug Resistant ◽

New Drugs ◽

Drug Resistant ◽

Adjuvant Effect ◽

Extensively Drug Resistant

Tuberculosis causes more than 1.2 million deaths each year. Worldwide, it is the first cause of death by a single infectious agent. The emergence of drug-resistant strains has limited pharmacological treatment of the disease and today, new drugs are urgently needed. Semi-synthetic mulinanes have previously shown important activity against multidrug-resistant (MDR) Mycobacterium tuberculosis. In this investigation, a new set of semi-synthetic mulinanes were synthetized, characterized, and evaluated for their in vitro activity against three drug-resistant clinical isolates of M. tuberculosis: MDR, pre-extensively Drug-Resistant (pre-XDR), and extensively Drug-Resistant (XDR), and against the drug-susceptible laboratory reference strain H37Rv. Derivative 1a showed the best anti-TB activity (minimum inhibitory concentration [MIC] = 5.4 µM) against the susceptible strain and was twice as potent (MIC = 2.7 µM) on the MDR, pre-XDR, and XDR strains and also possessed a bactericidal effect. Derivative 1a was also tested for its anti-TB activity in mice infected with the MDR strain. In this case, 1a produced a significant reduction of pulmonary bacilli loads, six times lower than the control, when tested at 0.2536 mg/Kg. In addition, 1a demonstrated an adjuvant effect by shortening second-line chemotherapy. Finally, the selectivity index of >15.64 shown by 1a when tested on Vero cells makes this derivative an important candidate for future studies in the development of novel antitubercular agents.

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Anti-mycobacterial Constituents from Medicinal Plants; A Review

Mini-Reviews in Medicinal Chemistry ◽

10.2174/1389557521666210525120325 ◽

2021 ◽

Vol 21 ◽

Author(s):

Nandan Sarkar ◽

Yadu Nandan Dey ◽

Dharmendra Kumar ◽

Mogana R

Keyword(s):

Mycobacterium Tuberculosis ◽

Medicinal Plants ◽

Development Process ◽

Multidrug Resistant ◽

Review Article ◽

New Drugs ◽

Drug Resistant ◽

Drug Discovery And Development ◽

Resistant Strains ◽

Drug Resistant Strains

: Effective treatment of tuberculosis has been hindered by the emergence of drug-resistant strains of Mycobacterium therapeutic facilities tuberculosis. With the global resurgence of tuberculosis with the development of multidrug-resistant cases, there is a call for the development of new drugs to combat these diseases. Throughout history, natural products have afforded a rich source of compounds that have found many applications in the fields of medicine, pharmacy and biology, and continued to play a significant role in the drug discovery and development process. This review article depicts the various potential plant extracts as well as plant-derived phytoconstituents against the H37rv, the most persistent strains of Mycobacterium tuberculosis and its multidrug strains.

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Functional Characterization by Genetic Complementation of aroB-Encoded Dehydroquinate Synthase from Mycobacterium tuberculosis H37Rv and Its Heterologous Expression and Purification

Journal of Bacteriology ◽

10.1128/jb.00425-07 ◽

2007 ◽

Vol 189 (17) ◽

pp. 6246-6252 ◽

Cited By ~ 13

Author(s):

Jordana Dutra de Mendonça ◽

Fernanda Ely ◽

Mario Sergio Palma ◽

Jeverson Frazzon ◽

Luiz Augusto Basso ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Targets ◽

Rational Design ◽

Functional Characterization ◽

Pcr Amplification ◽

Shikimate Pathway ◽

Aromatic Amino Acids ◽

Multidrug Resistant ◽

Genetic Complementation ◽

Dehydroquinate Synthase

ABSTRACT The recent recrudescence of Mycobacterium tuberculosis infection and the emergence of multidrug-resistant strains have created an urgent need for new therapeutics against tuberculosis. The enzymes of the shikimate pathway are attractive drug targets because this route is absent in mammals and, in M. tuberculosis, it is essential for pathogen viability. This pathway leads to the biosynthesis of aromatic compounds, including aromatic amino acids, and it is found in plants, fungi, bacteria, and apicomplexan parasites. The aroB-encoded enzyme dehydroquinate synthase is the second enzyme of this pathway, and it catalyzes the cyclization of 3-deoxy-d-arabino-heptulosonate-7-phosphate in 3-dehydroquinate. Here we describe the PCR amplification and cloning of the aroB gene and the overexpression and purification of its product, dehydroquinate synthase, to homogeneity. In order to probe where the recombinant dehydroquinate synthase was active, genetic complementation studies were performed. The Escherichia coli AB2847 mutant was used to demonstrate that the plasmid construction was able to repair the mutants, allowing them to grow in minimal medium devoid of aromatic compound supplementation. In addition, homogeneous recombinant M. tuberculosis dehydroquinate synthase was active in the absence of other enzymes, showing that it is homomeric. These results will support the structural studies with M. tuberculosis dehydroquinate synthase that are essential for the rational design of antimycobacterial agents.

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New drugs for the treatment of Mycobacterium tuberculosis infection

Biomedicine & Pharmacotherapy ◽

10.1016/j.biopha.2017.04.105 ◽

2017 ◽

Vol 91 ◽

pp. 546-558 ◽

Cited By ~ 19

Author(s):

Manaf AlMatar ◽

Husam AlMandeal ◽

Işıl Var ◽

Begüm Kayar ◽

Fatih Köksal

Keyword(s):

Mycobacterium Tuberculosis ◽

New Drugs ◽

Tuberculosis Infection ◽

Mycobacterium Tuberculosis Infection

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The Combination of Sulfamethoxazole, Trimethoprim, and Isoniazid or Rifampin Is Bactericidal and Prevents the Emergence of Drug Resistance in Mycobacterium tuberculosis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00832-12 ◽

2012 ◽

Vol 56 (10) ◽

pp. 5142-5148 ◽

Cited By ~ 28

Author(s):

Catherine Vilchèze ◽

William R. Jacobs

Keyword(s):

Drug Resistance ◽

Mycobacterium Tuberculosis ◽

Multidrug Resistant ◽

New Drugs ◽

Drug Resistant ◽

Content Type ◽

The Past ◽

Tb Treatment

ABSTRACTThe challenges of developing new drugs to treat tuberculosis (TB) are indicated by the relatively small number of candidates entering clinical trials in the past decade. To overcome these issues, we reexamined two FDA-approved antibacterial drugs, sulfamethoxazole (SMX) and trimethoprim (TMP), for use in TB treatment. SMX and TMP inhibit folic acid biosynthesis and are used in combination to treat infections of the respiratory, urinary, and gastrointestinal tracts. The MICs of SMX and TMP, alone and in combination, were determined for drug-susceptible, multidrug-resistant (MDR), and extensively drug-resistantMycobacterium tuberculosisstrains. While TMP alone was not effective againstM. tuberculosis, the combination of TMP and SMX was bacteriostatic againstM. tuberculosis. Surprisingly, the combination of SMX and TMP was also active against a subset of MDRM. tuberculosisstrains. Treatment ofM. tuberculosiswith TMP-SMX and a first-line anti-TB drug, either isoniazid or rifampin, was bactericidal, demonstrating that the combination of TMP and SMX with isoniazid or rifampin was not antagonistic. Moreover, the addition of SMX-TMP in combination with either isoniazid or rifampin also prevented the emergence of drug resistancein vitro. In conclusion, this study further illustrates the opportunity to reevaluate the activity of TMP-SMXin vivoto prevent the emergence of drug-resistantM. tuberculosis.

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Azetidines kill Mycobacterium tuberculosis without detectable resistance by blocking mycolate assembly

10.1101/2020.09.03.281170 ◽

2020 ◽

Author(s):

Alice Lanne ◽

Yixin Cui ◽

Edward Browne ◽

Philip G. E. Craven ◽

Nicholas J. Cundy ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Mode Of Action ◽

Drug Targets ◽

Cell Envelope ◽

Mycolic Acid ◽

New Drugs ◽

Drug Resistant ◽

Target Deconvolution ◽

Mdr Tb ◽

Mycobacterial Cell Wall

AbstractTuberculosis (TB) is the leading cause of global morbidity and mortality resulting from infectious disease, with over 10 million new cases and 1.5 million deaths in 2019. This global emergency is exacerbated by the emergence of multi-drug-resistant MDR-TB and extensively-drug-resistant XDR-TB, therefore new drugs and new drug targets are urgently required. From a whole-cell phenotypic screen a series of azetidines derivatives termed BGAz, that elicit potent bactericidal activity with MIC99 values <10 μM against drug-sensitive Mycobacterium tuberculosis and MDR-TB were identified. These compounds demonstrate no detectable drug resistance. Mode of action and target deconvolution studies suggest that these compounds inhibit mycobacterial growth by interfering with cell envelope biogenesis, specifically late-stage mycolic acid biosynthesis. Transcriptomic analysis demonstrates that the BGAz compounds tested display a mode of action distinct from existing mycobacterial cell-wall inhibitors. In addition, the compounds tested exhibit toxicological and PK/PD profiles that pave the way for their development as anti-tubercular chemotherapies.

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Structure-based drug repurposing to inhibit the DNA gyrase of Mycobacterium tuberculosis

Biochemical Journal ◽

10.1042/bcj20200462 ◽

2020 ◽

Vol 477 (21) ◽

pp. 4167-4190

Author(s):

Balasubramani GL ◽

Rinky Rajput ◽

Manish Gupta ◽

Pradeep Dahiya ◽

Jitendra K. Thakur ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Drug Repurposing ◽

Dna Gyrase ◽

Dna Supercoiling ◽

New Drugs ◽

Resonance Spectroscopy ◽

Drug Resistant ◽

Ic50 Values ◽

Resistant Strains ◽

Drug Resistant Strains

Drug repurposing is an alternative avenue for identifying new drugs to treat tuberculosis (TB). Despite the broad-range of anti-tubercular drugs, the emergence of multi-drug-resistant and extensively drug-resistant strains of Mycobacterium tuberculosis (Mtb) H37Rv, as well as the significant death toll globally, necessitates the development of new and effective drugs to treat TB. In this study, we have employed a drug repurposing approach to address this drug resistance problem by screening the drugbank database to identify novel inhibitors of the Mtb target enzyme, DNA gyrase. The compounds were screened against the ATPase domain of the gyrase B subunit (MtbGyrB47), and the docking results showed that echinacoside, doxorubicin, epirubicin, and idarubicin possess high binding affinities against MtbGyrB47. Comprehensive assessment using fluorescence spectroscopy, surface plasmon resonance spectroscopy (SPR), and circular dichroism (CD) titration studies revealed echinacoside as a potent binder of MtbGyrB47. Furthermore, ATPase, and DNA supercoiling assays exhibited an IC50 values of 2.1–4.7 µM for echinacoside, doxorubicin, epirubicin, and idarubicin. Among these compounds, the least MIC90 of 6.3 and 12 μM were observed for epirubicin and echinacoside, respectively, against Mtb. Our findings indicate that echinacoside and epirubicin targets mycobacterial DNA gyrase, inhibit its catalytic cycle, and retard mycobacterium growth. Further, these compounds exhibit potential scaffolds for optimizing novel anti-mycobacterial agents that can act on drug-resistant strains.

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THE BIOLOGY OF MYCOBACTERIUM TUBERCULOSIS INFECTION.

Mediterranean Journal of Hematology and Infectious Diseases ◽

10.4084/mjhid.2013.070 ◽

2013 ◽

Vol 5 (1) ◽

pp. e2013070 ◽

Cited By ~ 47

Author(s):

Giovanni Delogu ◽

Michela Sali ◽

Giovanni Fadda

Keyword(s):

Mycobacterium Tuberculosis ◽

Latent Infection ◽

Dynamic Equilibrium ◽

New Drugs ◽

Effective Control ◽

Mycobacterium Tuberculosis Infection ◽

Global Pandemic ◽

Recent Advancement ◽

Common Opinion ◽

And Control

Tuberculosis (TB) still poses a major threat to mankind and during the last thirty years we have seen a recrudescence of the disease even in countries where TB was thought to be conquered. It is common opinion that more effective control tools such as new diagnostics, a new vaccine and new drugs are urgently needed to control the global pandemic, though the so far insufficient understanding of the Mycobacterium tuberculosis (Mtb) mechanism of pathogenesis is a major obstacle for the development of these control tools. In this review, we will summarize the recent advancement in the understanding of Mtb biology and on the pathogenesis of Mtb infection with emphasis on latent infection, with the change in paradigm of the last few years where the dichotomy between latent and active disease has been reconsidered in favor of a dynamic equilibrium between the host and the bacilli, encompassing a continuous spectrum of conditions that has been named TB spectrum. Implications for the diagnosis and control of disease in certain population will also be discussed.

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Bengamides display potent activity against drug-resistant Mycobacterium tuberculosis

Scientific Reports ◽

10.1038/s41598-019-50748-2 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Diana H. Quan ◽

Gayathri Nagalingam ◽

Ian Luck ◽

Nicholas Proschogo ◽

Vijaykumar Pillalamarri ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

New Drugs ◽

Bioactive Molecules ◽

Aminopeptidase Activity ◽

Drug Resistant ◽

Lead Compounds ◽

New Class ◽

Tb Treatment ◽

Bioassay Guided Fractionation

Abstract Mycobacterium tuberculosis infects over 10 million people annually and kills more people each year than any other human pathogen. The current tuberculosis (TB) vaccine is only partially effective in preventing infection, while current TB treatment is problematic in terms of length, complexity and patient compliance. There is an urgent need for new drugs to combat the burden of TB disease and the natural environment has re-emerged as a rich source of bioactive molecules for development of lead compounds. In this study, one species of marine sponge from the Tedania genus was found to yield samples with exceptionally potent activity against M. tuberculosis. Bioassay-guided fractionation identified bengamide B as the active component, which displayed activity in the nanomolar range against both drug-sensitive and drug-resistant M. tuberculosis. The active compound inhibited in vitro activity of M. tuberculosis MetAP1c protein, suggesting the potent inhibitory action may be due to interference with methionine aminopeptidase activity. Tedania-derived bengamide B was non-toxic against human cell lines, synergised with rifampicin for in vitro inhibition of bacterial growth and reduced intracellular replication of M. tuberculosis. Thus, bengamides isolated from Tedania sp. show significant potential as a new class of compounds for the treatment of drug-resistant M. tuberculosis.

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