Novel Bacterial NAD
            <sup>+</sup>
            -Dependent DNA Ligase Inhibitors with Broad-Spectrum Activity and Antibacterial Efficacy
            <i>In Vivo</i>

Mills, Scott D.; Eakin, Ann E.; Buurman, Ed T.; Newman, Joseph V.; Gao, Ning; Huynh, Hoan K.; Johnson, Kenneth D.; Lahiri, S. C.; Shapiro, Adam B.; Walkup, Grant K.; Yang, Wei; Stokes, Suzanne S.

doi:10.1128/aac.01181-10

Cited by 77 publications

(100 citation statements)

References 36 publications

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“…aureus (MIC 32-64 mg l 21 ). The antibacterial activity of several 2-substituted adenosine analogues has recently been reported by Mills et al (2011). This study showed that 2-thio-n-butyl adenosine was an inhibitor of the bacterial NAD + -dependent DNA ligase.…”

Section: Resultsmentioning

confidence: 78%

“…Some recent experiences, however, have proven the great value of this field of research. For example, complex nucleosides exhibit antibacterial activity by specific inhibition of cell-wall peptidoglycan biosynthesis (Kimura & Bugg, 2003), certain bacterial enzymes, such as purine nucleoside phosphorylases (Bzowska et al, 2000), and DNA ligases (Mills et al, 2011). In particular, adenosine analogues such as cordycepin (Ahn et al, 2000), 29-amino-29-deoxyadenosine and oxetanocin (Shimada et al, 1986), formycins (Bzowska et al, 2000), toyocamycin and its derivatives (Kimura & Bugg, 2003) and some purine derivatives (Tunçbilek et al, 2009) showed biological activity including antibiotic properties.…”

Section: Introductionmentioning

confidence: 99%

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In vitro antibacterial activity of different adenosine analogues

Vitali

Petrelli

Lambertucci

et al. 2012

Journal of Medical Microbiology

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Nucleoside analogues may represent good candidates for the discovery of new antibacterial agents, therefore, a library of adenosine analogues was assessed for their antibacterial activity, and the relationship between the structure and activity of these molecules was outlined. Antibacterial activity was evaluated against that of reference strains of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Escherichia coli and Pseudomonas aeruginosa. We tested 54 adenosine analogues, modified both at ribose and base moieties, including adenine and 1/3-deazaadenine derivatives substituted in the 2-and/or N 6 -positions and bearing N-9 sugar moieties, such as ribose, 29-deoxyribose, 39-deoxyribose, 29,39-dideoxyribose or cycloalkyl groups like cyclopentane. The data obtained, MIC and minimal bactericidal concentrations demonstrated that the presence of bulky substituents such as cycloheptyl and cyclooctyl rings on the N 6 -amino, together with a chlorine atom in the 2-position, conferred antibacterial activity against the Gram-positive group with MIC values ranging from 16 to 128 mg l "1 . The intact sugar moiety seemed to be not essential for antimicrobial activity and nucleosides bearing deoxyribose or cyclopentyl groups associated with bulky substituents in N 6 -position showed good antimicrobial properties. Furthermore, N-1 proved to be non-crucial and the 2-chloro-N 6 -cyclooctyl-1-deaza-39-deoxyadenosine and 29,39-dideoxyadenosine compounds were among the more active in the series with an MIC of 32 mg l "1 against Staph. aureus and Strep. pneumoniae. None of the analogues was active against the two Gram-negative species tested. Hence, adenosine derivatives bearing bulky substituents in the N 6 -position may represent good lead compounds for the future discovery of a novel series of antibacterial agents.

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Section: Resultsmentioning

confidence: 78%

Section: Introductionmentioning

confidence: 99%

In vitro antibacterial activity of different adenosine analogues

Vitali

Petrelli

Lambertucci

et al. 2012

Journal of Medical Microbiology

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“…the assay has been used for high-throughput screening for the discovery of bacterial NAD + -dependent DNA ligase inhibitors. 5 this article describes in greater detail the development and principle of the assay and illustrates its use for measuring the steady-state kinetic constants of Haemophilus influenzae NAD + -dependent DNA ligase and of inhibitor potency.…”

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confidence: 99%

A High-Throughput Fluorescence Resonance Energy Transfer-Based Assay for DNA Ligase

et al. 2011

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DNA ligase is the enzyme that catalyzes the formation of the backbone phosphodiester bond between the 5′-PO4 and 3′-OH of adjacent DNA nucleotides at single-stranded nicks. These nicks occur between Okazaki fragments during replication of the lagging strand of the DNA as well as during DNA repair and recombination. As essential enzymes for DNA replication, the NAD+-dependent DNA ligases of pathogenic bacteria are potential targets for the development of antibacterial drugs. For the purposes of drug discovery, a high-throughput assay for DNA ligase activity is invaluable. This article describes a straightforward, fluorescence resonance energy transfer–based DNA ligase assay that is well suited for high-throughput screening for DNA ligase inhibitors as well as for use in enzyme kinetics studies. Its use is demonstrated for measurement of the steady-state kinetic constants of Haemophilus influenzae NAD+-dependent DNA ligase and for measurement of the potency of an inhibitor of this enzyme.

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“…HY-18649A; Medchemexpress LLC, Princeton, NJ, USA), used as a control] was measured at 10 different compound concentrations in the reaction system in order to determine the 50% inhibitory concentration (IC 50 ). Assays for the other bacterial LigA isozymes were performed as described above for H. influenzae LigA, with the following changes in order to optimize enzyme performance, as previously described (22,23 …”

Section: Methodsmentioning

confidence: 99%

Separation of cordycepin from Cordyceps militaris fermentation supernatant using preparative HPLC and evaluation of its antibacterial activity as an NAD+-dependent DNA ligase inhibitor

Zhou

Cai

et al. 2016

Experimental and Therapeutic Medicine

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Abstract. Cordycepin exhibits various bio-activities, including anticancer, antibacterial, antiviral and immune regulation activities, and is a significant focus of research. However, the preparation of high-purity cordycepin remains challenging. Also, the molecular target with which cordycepin interacts to cause an antibacterial effect remains unknown. In the present study, cordycepin was prepared by preparative high-performance liquid chromatography (prep-HPLC) and the purity obtained was 99.6%, indicating that this technique may be useful for the large-scale isolation of cordycepin in the future. The results of computational molecular docking analysis indicated that the interaction energy between cordycepin and NAD + -dependent DNA ligase (LigA) was lower than that between cordycepin and other common antibacterial targets. The highly pure cordycepin obtained by prep-HPLC demonstrated inhibitory activity against LigA from various bacteria in vitro. In conclusion, cordycepin may be useful as a broad-spectrum antibiotic targeting LigA in various bacteria. IntroductionCordycepin is an antimicrobial substance that was first isolated from the entomopathogenic fungus Cordyceps militaris by Cunningham et al in 1950 (1). Since its discovery, a number of studies have focused on this substance (2,3). Cordycepin ( Fig. 1A) with the molecular formula C 10 H 13 N 5 O 3 has been demonstrated to have various bio-activities, including anticancer (4-7), antibacterial (8), antiviral (9,10) and immune regulation activities (11) and has became a topic of interest to various research groups.However, highly pure cordycepin is challenging to obtain; various drawbacks have been identified in the purification techniques of ion resin adsorption (12,13), silica gel column chromatography (14) and supercritical extraction (15). The high price of cordycepin acts as a significant barrier to the study of its bio-activity. Preparative high performance liquid chromatography (prep-HPLC) can shorten the production cycle, has relatively low operating costs, and is simple to carry out. Therefore, it is commonly used for the purification of chemical drugs and biological drugs by pharmaceutical companies and particularly in industrial purification (16). Prep-HPLC can be used to provide purified samples on a kilogram, or even ton, scale. Also, prep-HPLC is an effective technology currently used for natural product isolation in industry (17). The present study describes a new method of obtaining high-purity cordycepin by prep-HPLC, which may be useful for the large-scale isolation of cordycepin in the future.Although studies have shown that cordycepin is a broad-spectrum antibiotic (18), studies focusing on the mechanism of the antibacterial activity and the possible targets are lacking. Molecular modeling and docking technology, which is efficient, accurate and simple to use, can reproduce the true interaction process between drug and targets that some of the current laboratory techniques are unable to detect, and is a method for predicting the mecha...

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Novel Bacterial NAD ⁺ -Dependent DNA Ligase Inhibitors with Broad-Spectrum Activity and Antibacterial Efficacy In Vivo

Cited by 77 publications

References 36 publications

In vitro antibacterial activity of different adenosine analogues

In vitro antibacterial activity of different adenosine analogues

A High-Throughput Fluorescence Resonance Energy Transfer-Based Assay for DNA Ligase

Separation of cordycepin from Cordyceps militaris fermentation supernatant using preparative HPLC and evaluation of its antibacterial activity as an NAD+-dependent DNA ligase inhibitor

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Novel Bacterial NAD + -Dependent DNA Ligase Inhibitors with Broad-Spectrum Activity and Antibacterial Efficacy In Vivo

Cited by 77 publications

References 36 publications

In vitro antibacterial activity of different adenosine analogues

In vitro antibacterial activity of different adenosine analogues

A High-Throughput Fluorescence Resonance Energy Transfer-Based Assay for DNA Ligase

Separation of cordycepin from Cordyceps militaris fermentation supernatant using preparative HPLC and evaluation of its antibacterial activity as an NAD+-dependent DNA ligase inhibitor

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Novel Bacterial NAD ⁺ -Dependent DNA Ligase Inhibitors with Broad-Spectrum Activity and Antibacterial Efficacy In Vivo