Anti-tubercular activity of a natural stilbene and its synthetic derivatives

id000036 10.3205/id000036 urn:nbn:de:0183-id0000360 Research Article Anti-tubercular activity of a natural stilbene and its synthetic derivatives Reinheimer Reinheimer Claudia C Priv.-Doz. Dr. Dr. med.

Institute of Medical Microbiology and Infection Control, Hospital of Goethe-University, Paul-Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany, Phone: +49-69-6301-6282Institute of Medical Microbiology and Infection Control, Hospital of Goethe-University, Frankfurt, Germany

claudia.reinheimer@kgu.de author Büttner Büttner Dominik D

Institute of Pharmaceutical Chemistry, Goethe-University, Frankfurt, Germany

author Proschak Proschak Eugen E

Institute of Pharmaceutical Chemistry, Goethe-University, Frankfurt, Germany

author Bode Bode Helge B. HB

Merck endowed chair for Molecular Biotechnology, Department of Biosciences and Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-University, Frankfurt, Germany

author Kempf Kempf Volkhard A. J. VAJ

Institute of Medical Microbiology and Infection Control, Hospital of Goethe-University, Frankfurt, Germany

author Wichelhaus Wichelhaus Thomas A. TA

Institute of Medical Microbiology and Infection Control, Hospital of Goethe-University, Frankfurt, Germany

author German Medical Science GMS Publishing House

Düsseldorf

610 tuberculosis drug resistance new substances epoxide hydrolases stilbene 20180201 engl This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). 2195-8831 6 GMS Infectious Diseases GMS Infect Dis 01 Objectives: Tuberculosis (TB) and multidrug- and extensively drug-resistant TB in particular are remaining a major global health challenge and efficient new drugs against TB are needed. This study evaluated the anti-tubercular activity of a natural stilbene and its synthetic derivatives against M. tuberculosis.Methods: Isopropylstilbene and its synthetic derivatives were analyzed for their anti-tubercular activity against M. tuberculosis ATCC 27294 as well as multidrug- and extensively drug-resistant M. tuberculosis clinical isolates by using MGIT 960 instrumentation and EpiCenter software equipped with TB eXiST module. Cytotoxic effects of drug candidates were determined by a MTT dye reduction assay using A549 adenocarcinomic human alveolar basal epithelial cells.Results: Growth of M. tuberculosis ATCC 27294 was suppressed by the natural isopropylstilbene HB64 as well as synthetic derivatives DB56 and DB55 at 25 µg/ml. Growth of clinical isolates MDR and XDR M. tuberculosis was suppressed by HB64 at 100 µg/ml as well as by synthetic derivatives DB56 and DB55 at 50 µg/ml and 25 µg/ml, respectively. No anti-tubercular activity was demonstrated for synthetic derivatives DB53, EB251, and RB57 at 100 µg/ml. Toxicity in terms of IC50 values of HB64, DB55 and DB56 were 7.92 µg/ml, 12.15 µg/ml and 16.01 µg/ml, respectively.Conclusions: Synthetical derivatives of stilbene might be effective candidates as anti-tubercular drugs. However, toxicity of these substances as determined by IC50 values might limit therapeutic success in vivo. Further investigations should address lowering the toxicity for parenteral administration by remodeling stilbene derivatives. IntroductionTuberculosis (TB) incidence is declining globally, however, multidrug-resistant TB (MDR-TB), defined as TB with resistance to isoniazid (INH) and rifampicin (R), and extensively drug-resistant (XDR-TB), defined as MDR with additional resistance to a fluoroquinolone and to one or more of the injectable drugs, remain major global health challenges . MDR-TB strains are representing a major threat to TB control since the 1990s, and the emergence of the first XDR-TB cases in 2006 has impressively demonstrated that treatment options for XDR-TB have become alarmingly narrow and the need for new drugs is pressing. While evidence supports the repurposing of antibiotics approved for other indications, such as linezolid , the new drugs bedaquiline and delamanid have recently been approved for the treatment of drug-resistant tuberculosis . Despite this silver lining, the increasing burden of XDR-TB underlines the demand on efficient new substances. Since the sequence of the M. tuberculosis genome has been published, a number of characteristics have been described, e.g. the genome encodes for at least 20 epoxide hydrolases (EHs), which has formerly been reported by Cole et al. to be an unusually large number for a single bacterium . EHs are key enzymes in the arachidonic acid cascade and essential to many organisms because of their ability to transform chemically reactive and detrimental epoxides into less reactive substances , . Substances to inhibit EHs may therefore promising substances in future anti-tubercular therapy. Stilbenes have formerly been observed to have an inhibitory effect on EHs . Natural stilbenes are a group of nonflavonoid phytochemicals of polyphenolic structure characterized by the presence of a 1,2-diphenylethylene core , . Considering (a) the stilbenes’ inhibitory effect on EHs, (b) the potential toxicity of EH substrates to M. tuberculosis in general and (c) the unusual large number of EHs in M. tuberculosis, EHs therefore are proposed to be a promising target for innovative anti-tubercular drugs (ATD). The role of stilbenes and stilbene-derived substances as potentially innovative ATD has scientifically not been addressed yet, to our knowledge. This study therefore aims at evaluation the anti-tubercular activity of a natural stilbene and its synthetic derivatives against M. tuberculosis. Material and methodsNatural stilbenes and its synthetic derivativesThe natural stilbene isopropylstilbene (HB64, Figure 1a ) isolated from Photorhabdus luminescens , as well as its synthetic derivatives were analyzed for their anti-tubercular activity.(E)-styryl-1H-benzo[d]imidazoles (DB53, DB55, DB56)In a round-bottom flask 250 mg of 3,4-diaminotoluene derivatives (1.0 eq), o-trifluoromethyl cinnamic acid (1.0 eq</PlainText></TextGroup>) and EDC*HCl (1.5 eq) was dissolved in 10 ml DMF. Catalytic amounts of DMAP (0.05 eq) and imidazole (0.05 eq) were added, and the reaction was stirred for <TextGroup><PlainText>4 h</PlainText></TextGroup> at room temperature. After addition of 150 ml of distilled water, the precipitated solid was obtained by vacuum filtration and washed twice with 10 ml of water.</Pgraph><Pgraph>100 mg of cinnamic acid amides were suspended in <TextGroup><PlainText>2 ml</PlainText></TextGroup> of 6 M aqueous hydrochloric acid. 2.5 ml of EtOH were added, and the mixture was heated under microwave irradiation for 10 minutes at 120°C. The reaction was cooled to room temperature, and subsequently cooled at 4°C. The resulting sediment was filtered off and washed with water. Substances are named DB53 (R=F), DB55 (R=CH<Subscript>3</Subscript>), DB56 (R=Cl) (Figure 1b <ImgLink imgNo="1" imgType="figure"/>).</Pgraph><SubHeadline2>N-((3s, 5s, 7s)-adamantam-1-yl)-6-chloro-2-(4-morpholinophenyl)imidazo[1, 2-a]pyridine-3-amine (EB251) and ethyl 1-(3-((3-cyclohexylureido)methyl)benzyl)-1H-pyrrole-2-carboxylate (RB57)</SubHeadline2><Pgraph>Synthesis has formerly been described by Buscató et al. <TextLink reference="14"></TextLink>, <TextLink reference="15"></TextLink> (Figure 1c,d <ImgLink imgNo="1" imgType="figure"/>).</Pgraph><Pgraph> </Pgraph><SubHeadline>Bacterial strains</SubHeadline><Pgraph>Reference strain <Mark2>M. tuberculosis</Mark2> ATCC 27294 as well as MDR and XDR <Mark2>M. tuberculosis</Mark2> clinical isolates were used to determine the anti-tubercular activity of a natural stilbene and its synthetical derivatives. </Pgraph><SubHeadline>Susceptibility testing of M. tuberculosis </SubHeadline><Pgraph>The MGIT 960 system was used for anti-tubercular susceptibility testing as previously described <TextLink reference="16"></TextLink>. MGIT tubes supplemented with 0.8 ml of supplement (MGIT 960 SIRE supplement; Becton Dickinson) were inoculated with <TextGroup><PlainText>0.1 ml</PlainText></TextGroup> of the drug solution at various concentrations and 0.5 ml of the test strain suspension, i.e. <Mark2>M. tuberculosis</Mark2> ATCC 27294 as well as MDR and XDR <Mark2>M. tuberculosis</Mark2> clinical isolates. For preparation of the drug-free growth control tube, the organism suspension was diluted 1:100 with sterile saline, and then 0.5 ml was inoculated into the tube (proportion testing). Quantitative drug susceptibility testing (DST) was performed using the MGIT 960 instrumentation and EpiCenter software version 5.53 equipped with the TB eXiST module (Becton Dickinson), providing features as previously described <TextLink reference="16"></TextLink>, including automated recording of the readings, additional incubation time beyond the time to positivity of the drug-free control, and graphical representation of the growth unit (GU) value increase. The susceptibility testing sets were placed in the MGIT 960 instrument and continuously monitored using EpiCenter TB eXiST software.</Pgraph><Pgraph>The susceptibility testing was performed in triplicate. Results were interpreted as follows: at the time when the growth unit (GU) of the drug-free control tube was >400, if the GU of the drug-containing tube to be compared was ≥100, the strain was resistant (R). If the GU of the drug-containing tube was <100, it was interpreted as susceptible (S). Rifampicin and isoniazid were used as positive control drugs.</Pgraph><SubHeadline>Assessment of cytotoxic effects</SubHeadline><Pgraph>Cytotoxic effects of drug candidates were determined <TextGroup><PlainText>by a</PlainText></TextGroup> 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye reduction assay. Cells were plated in 96-well microtiter plates at a density 20,000–50,000 cells/ well <TextLink reference="17"></TextLink>. The cells were incubated in a culture medium at concentrations between 0.19 and 100 µg/ml of the anti-tubercular drug. After 5 days of incubation at 37.0°C in 5% CO<Subscript>2</Subscript> atmosphere, 25 µl MTT substrate (2 mg/ml) was added and plates were incubated at 37.0°C for <TextGroup><PlainText>4 hours</PlainText></TextGroup>. After incubation, cells were lysed in 100 ml buffer containing 2% (w/v) SDS and 5<TextGroup><PlainText>0% (v</PlainText></TextGroup>/v) <Mark2>N,N</Mark2>-dimethylformamide with the pH adjusted to 4.7. Absorbance at 560/620 nm was determined for each well using a 96-well multiscanner. After correcting for the background, the results were expressed as percentage viability relative to a control culture which received no drug.</Pgraph></TextBlock> <TextBlock linked="yes" name="Results and discussion"> <MainHeadline>Results and discussion</MainHeadline><Pgraph>The global spread of TB and its multi- or extensively drug resistant strains is remaining a matter of public health concern worldwide <TextLink reference="1"></TextLink>, <TextLink reference="2"></TextLink>. Therefore, new compounds against <Mark2>M. tuberculosis</Mark2> are urgently needed <TextLink reference="18"></TextLink>. In order to address this issue, we tested the anti-tubercular activity of a natural stilbene and its synthetic derivatives.</Pgraph><Pgraph>Numerous assay systems have been established for the evaluation of compounds against <Mark2>M. tuberculosis</Mark2> and we followed a method based on monitoring growth/inhibition via fluorometric detection of oxygen consumption <TextLink reference="16"></TextLink>.</Pgraph><Pgraph>A total of six substances were used for quantitative DST on the basis of the MGIT 960 instrumentation and <TextGroup><PlainText>EpiCenter</PlainText></TextGroup>. Growth of <Mark2>M. tuberculosis</Mark2> ATCC 27294 was suppressed by HB64 at 25 µg/ml as well as synthetic derivatives DB56 and DB55 at 25 µg/ml each (Table 1 <ImgLink imgNo="1" imgType="table"/>). Growth of clinical isolate MDR <Mark2>M. tuberculosis</Mark2> was suppressed by HB64 at 100 µg/ml as well as by synthetic derivatives DB56 and DB55 at 50 µg/ml and 25 µg/ml, respectively (Table 1 <ImgLink imgNo="1" imgType="table"/>). Furthermore, HB64, DB55 and DB56 were tested against clinical isolate XDR <TextGroup><Mark2>M. tuberculosis</Mark2></TextGroup> with growth suppression at 100 µg/ml, 25 µg/ml and 50 µg/ml, respectively (Table 1 <ImgLink imgNo="1" imgType="table"/>). Interestingly, compared with the natural stilbene, synthetic derivatives (DB55 and DB56) showed enhanced <Mark2>in vitro</Mark2> activity against MDR and XDR <Mark2>M. tuberculosis</Mark2>.</Pgraph><Pgraph>The natural stilbene (HB64) also known as benvitimod has been accepted for clinical trial for the treatment of psoriasis. Benvitimod demonstrated to be an effective topical anti-inflammatory molecule that showed a negligible absorption and was well tolerated <TextLink reference="19"></TextLink>. In the present study we determined the toxicity in terms of IC<Subscript>50</Subscript> values for those substances that show anti-tubercular activity (Table 2 <ImgLink imgNo="2" imgType="table"/>). The IC<Subscript>50</Subscript> values turned out to be lower than the MIC of those substances exhibiting anti-tubercular activity, an issue that would limit systemic application <Mark2>in vivo</Mark2>. Furthermore, rapid metabolization of stilbenes after administration has been described <TextLink reference="20"></TextLink>, which might further influence pharmacokinetics and -activities.</Pgraph><Pgraph>Finally, synthetical derivatives of stilbene might be effective candidates as anti-tubercular drugs. However, toxicity of these substances as determined by IC<Subscript>50</Subscript> values limit therapeutic success <Mark2>in vivo</Mark2>. Further investigations need to address lowering the toxicity by remodeling stilbene derivatives.</Pgraph></TextBlock> <TextBlock linked="yes" name="Notes"> <MainHeadline>Notes</MainHeadline><SubHeadline>Competing interests</SubHeadline><Pgraph>The authors declare that they have no competing <TextGroup><PlainText>interests</PlainText></TextGroup>.</Pgraph><SubHeadline>Acknowledgments</SubHeadline><Pgraph>We thank Denia Frank and Kuflom Gebreamlack for excellent technical support.</Pgraph><SubHeadline>Authors’ contributions</SubHeadline><Pgraph><UnorderedList><ListItem level="1">Conceptualization: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Formal analysis: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Funding acquisition: CR, TW</ListItem><ListItem level="1">Investigation: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Methodology: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Project administration: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Resources: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Software: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Supervision: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Validation: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Visualization: CR, DB, EP, HB, VK, TW</ListItem><ListItem level="1">Writing – original draft: CR, TW</ListItem><ListItem level="1">Writing – review & editing: CR, DB, EP, HB, VK, TW</ListItem></UnorderedList></Pgraph><SubHeadline>Funding</SubHeadline><Pgraph>This work was supported by the Foundation <Mark2>Nachlässe Marie Christine Held und Erika Hecker,</Mark2> Hospital of Goethe-University, Frankfurt am Main, Germany.</Pgraph></TextBlock> <References linked="yes"> <Reference refNo="1"> <RefAuthor>Diel R</RefAuthor> <RefAuthor>Nienhaus A</RefAuthor> <RefAuthor>Lampenius N</RefAuthor> <RefAuthor>Rüsch-Gerdes S</RefAuthor> <RefAuthor>Richter E</RefAuthor> <RefTitle>Cost of multi drug resistance tuberculosis in Germany</RefTitle> <RefYear>2014</RefYear> <RefJournal>Respir Med</RefJournal> <RefPage>1677-87</RefPage> <RefTotal>Diel R, Nienhaus A, Lampenius N, Rüsch-Gerdes S, Richter E. Cost of multi drug resistance tuberculosis in Germany. Respir Med. 2014 Nov;108(11):1677-87. DOI: 10.1016/j.rmed.2014.09.021</RefTotal> <RefLink>http://dx.doi.org/10.1016/j.rmed.2014.09.021</RefLink> </Reference> <Reference refNo="2"> <RefAuthor>Centers for Disease Control and Prevention (CDC)</RefAuthor> <RefTitle>Emergence of Mycobacterium tuberculosis with extensive resistance to second-line drugs – worldwide, 2000-2004</RefTitle> <RefYear>2006</RefYear> <RefJournal>MMWR Morb Mortal Wkly Rep</RefJournal> <RefPage>301-5</RefPage> <RefTotal>Centers for Disease Control and Prevention (CDC). Emergence of Mycobacterium tuberculosis with extensive resistance to second-line drugs – worldwide, 2000-2004. MMWR Morb Mortal Wkly Rep. 2006 Mar;55(11):301-5.</RefTotal> </Reference> <Reference refNo="3"> <RefAuthor>Cox H</RefAuthor> <RefAuthor>Hughes J</RefAuthor> <RefAuthor>Daniels J</RefAuthor> <RefAuthor>Azevedo V</RefAuthor> <RefAuthor>McDermid C</RefAuthor> <RefAuthor>Poolman M</RefAuthor> <RefAuthor>Boulle A</RefAuthor> <RefAuthor>Goemaere E</RefAuthor> <RefAuthor>van Cutsem G</RefAuthor> <RefTitle>Community-based treatment of drug-resistant tuberculosis in Khayelitsha, South Africa</RefTitle> <RefYear>2014</RefYear> <RefJournal>Int J Tuberc Lung Dis</RefJournal> <RefPage>441-8</RefPage> <RefTotal>Cox H, Hughes J, Daniels J, Azevedo V, McDermid C, Poolman M, Boulle A, Goemaere E, van Cutsem G. Community-based treatment of drug-resistant tuberculosis in Khayelitsha, South Africa. Int J Tuberc Lung Dis. 2014 Apr;18(4):441-8. DOI: 10.5588/ijtld.13.0742</RefTotal> <RefLink>http://dx.doi.org/10.5588/ijtld.13.0742</RefLink> </Reference> <Reference refNo="4"> <RefAuthor>Reed C</RefAuthor> <RefAuthor>Mason L</RefAuthor> <RefAuthor>Cox H</RefAuthor> <RefAuthor>Seaworth B</RefAuthor> <RefAuthor>Lessem E</RefAuthor> <RefAuthor>Furin J</RefAuthor> <RefTitle>Compassionate and optimum use of new tuberculosis drugs</RefTitle> <RefYear>2015</RefYear> <RefJournal>Lancet Infect Dis</RefJournal> <RefPage>1131</RefPage> <RefTotal>Reed C, Mason L, Cox H, Seaworth B, Lessem E, Furin J. Compassionate and optimum use of new tuberculosis drugs. Lancet Infect Dis. 2015 Oct;15(10):1131. DOI: 10.1016/S1473-3099(15)00294-7</RefTotal> <RefLink>http://dx.doi.org/10.1016/S1473-3099(15)00294-7</RefLink> </Reference> <Reference refNo="5"> <RefAuthor>Cole ST</RefAuthor> <RefAuthor>Brosch R</RefAuthor> <RefAuthor>Parkhill J</RefAuthor> <RefAuthor>Garnier T</RefAuthor> <RefAuthor>Churcher C</RefAuthor> <RefAuthor>Harris D</RefAuthor> <RefAuthor>Gordon SV</RefAuthor> <RefAuthor>Eiglmeier K</RefAuthor> <RefAuthor>Gas S</RefAuthor> <RefAuthor>Barry CE 3rd</RefAuthor> <RefAuthor>Tekaia F</RefAuthor> <RefAuthor>Badcock K</RefAuthor> <RefAuthor>Basham D</RefAuthor> <RefAuthor>Brown D</RefAuthor> <RefAuthor>Chillingworth T</RefAuthor> <RefAuthor>Connor R</RefAuthor> <RefAuthor>Davies R</RefAuthor> <RefAuthor>Devlin K</RefAuthor> <RefAuthor>Feltwell T</RefAuthor> <RefAuthor>Gentles S</RefAuthor> <RefAuthor>Hamlin N</RefAuthor> <RefAuthor>Holroyd S</RefAuthor> <RefAuthor>Hornsby T</RefAuthor> <RefAuthor>Jagels K</RefAuthor> <RefAuthor>Krogh A</RefAuthor> <RefAuthor>McLean J</RefAuthor> <RefAuthor>Moule S</RefAuthor> <RefAuthor>Murphy L</RefAuthor> <RefAuthor>Oliver K</RefAuthor> <RefAuthor>Osborne J</RefAuthor> <RefAuthor>Quail MA</RefAuthor> <RefAuthor>Rajandream MA</RefAuthor> <RefAuthor>Rogers J</RefAuthor> <RefAuthor>Rutter S</RefAuthor> <RefAuthor>Seeger K</RefAuthor> <RefAuthor>Skelton J</RefAuthor> <RefAuthor>Squares R</RefAuthor> <RefAuthor>Squares S</RefAuthor> <RefAuthor>Sulston JE</RefAuthor> <RefAuthor>Taylor K</RefAuthor> <RefAuthor>Whitehead S</RefAuthor> <RefAuthor>Barrell BG</RefAuthor> <RefTitle>Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence</RefTitle> <RefYear>1998</RefYear> <RefJournal>Nature</RefJournal> <RefPage>537-44</RefPage> <RefTotal>Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon SV, Eiglmeier K, Gas S, Barry CE 3rd, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Krogh A, McLean J, Moule S, Murphy L, Oliver K, Osborne J, Quail MA, Rajandream MA, Rogers J, Rutter S, Seeger K, Skelton J, Squares R, Squares S, Sulston JE, Taylor K, Whitehead S, Barrell BG. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998 Jun 11;393(6685):537-44. Erratum in: Nature 1998 Nov 12;396(6707):190. DOI: 10.1038/31159</RefTotal> <RefLink>http://dx.doi.org/10.1038/31159</RefLink> </Reference> <Reference refNo="6"> <RefAuthor>Imig JD</RefAuthor> <RefAuthor>Hammock BD</RefAuthor> <RefTitle>Soluble epoxide hydrolase as a therapeutic target for cardiovascular diseases</RefTitle> <RefYear>2009</RefYear> <RefJournal>Nat Rev Drug Discov</RefJournal> <RefPage>794-805</RefPage> <RefTotal>Imig JD, Hammock BD. Soluble epoxide hydrolase as a therapeutic target for cardiovascular diseases. Nat Rev Drug Discov. 2009 Oct;8(10):794-805. DOI: 10.1038/nrd2875</RefTotal> <RefLink>http://dx.doi.org/10.1038/nrd2875</RefLink> </Reference> <Reference refNo="7"> <RefAuthor>Morisseau C</RefAuthor> <RefTitle>Role of epoxide hydrolases in lipid metabolism</RefTitle> <RefYear>2013</RefYear> <RefJournal>Biochimie</RefJournal> <RefPage>91-5</RefPage> <RefTotal>Morisseau C. Role of epoxide hydrolases in lipid metabolism. Biochimie. 2013 Jan;95(1):91-5. DOI: 10.1016/j.biochi.2012.06.011</RefTotal> <RefLink>http://dx.doi.org/10.1016/j.biochi.2012.06.011</RefLink> </Reference> <Reference refNo="8"> <RefAuthor>Arand M</RefAuthor> <RefAuthor>Hallberg BM</RefAuthor> <RefAuthor>Zou J</RefAuthor> <RefAuthor>Bergfors T</RefAuthor> <RefAuthor>Oesch F</RefAuthor> <RefAuthor>van der Werf MJ</RefAuthor> <RefAuthor>de Bont JA</RefAuthor> <RefAuthor>Jones TA</RefAuthor> <RefAuthor>Mowbray SL</RefAuthor> <RefTitle>Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site</RefTitle> <RefYear>2003</RefYear> <RefJournal>EMBO J</RefJournal> <RefPage>2583-92</RefPage> <RefTotal>Arand M, Hallberg BM, Zou J, Bergfors T, Oesch F, van der Werf MJ, de Bont JA, Jones TA, Mowbray SL. Structure of Rhodococcus erythropolis limonene-1,2-epoxide hydrolase reveals a novel active site. EMBO J. 2003 Jun;22(11):2583-92. DOI: 10.1093/emboj/cdg275</RefTotal> <RefLink>http://dx.doi.org/10.1093/emboj/cdg275</RefLink> </Reference> <Reference refNo="9"> <RefAuthor>Buscató E</RefAuthor> <RefAuthor>Büttner D</RefAuthor> <RefAuthor>Brüggerhoff A</RefAuthor> <RefAuthor>Klingler FM</RefAuthor> <RefAuthor>Weber J</RefAuthor> <RefAuthor>Scholz B</RefAuthor> <RefAuthor>Zivković A</RefAuthor> <RefAuthor>Marschalek R</RefAuthor> <RefAuthor>Stark H</RefAuthor> <RefAuthor>Steinhilber D</RefAuthor> <RefAuthor>Bode HB</RefAuthor> <RefAuthor>Proschak E</RefAuthor> <RefTitle>From a multipotent stilbene to soluble epoxide hydrolase inhibitors with antiproliferative properties</RefTitle> <RefYear>2013</RefYear> <RefJournal>ChemMedChem</RefJournal> <RefPage>919-23</RefPage> <RefTotal>Buscató E, Büttner D, Brüggerhoff A, Klingler FM, Weber J, Scholz B, Zivković A, Marschalek R, Stark H, Steinhilber D, Bode HB, Proschak E. From a multipotent stilbene to soluble epoxide hydrolase inhibitors with antiproliferative properties. ChemMedChem. 2013 Jun;8(6):919-23. DOI: 10.1002/cmdc.201300057</RefTotal> <RefLink>http://dx.doi.org/10.1002/cmdc.201300057</RefLink> </Reference> <Reference refNo="10"> <RefAuthor>Shen T</RefAuthor> <RefAuthor>Wang XN</RefAuthor> <RefAuthor>Lou HX</RefAuthor> <RefTitle>Natural stilbenes: an overview</RefTitle> <RefYear>2009</RefYear> <RefJournal>Nat Prod Rep</RefJournal> <RefPage>916-35</RefPage> <RefTotal>Shen T, Wang XN, Lou HX. Natural stilbenes: an overview. Nat Prod Rep. 2009 Jul;26(7):916-35. DOI: 10.1039/b905960a</RefTotal> <RefLink>http://dx.doi.org/10.1039/b905960a</RefLink> </Reference> <Reference refNo="11"> <RefAuthor>Rivière C</RefAuthor> <RefAuthor>Pawlus AD</RefAuthor> <RefAuthor>Mérillon JM</RefAuthor> <RefTitle>Natural stilbenoids: distribution in the plant kingdom and chemotaxonomic interest in Vitaceae</RefTitle> <RefYear>2012</RefYear> <RefJournal>Nat Prod Rep</RefJournal> <RefPage>1317-33</RefPage> <RefTotal>Rivière C, Pawlus AD, Mérillon JM. Natural stilbenoids: distribution in the plant kingdom and chemotaxonomic interest in Vitaceae. Nat Prod Rep. 2012 Nov;29(11):1317-33. DOI: 10.1039/c2np20049j</RefTotal> <RefLink>http://dx.doi.org/10.1039/c2np20049j</RefLink> </Reference> <Reference refNo="12"> <RefAuthor>Kronenwerth M</RefAuthor> <RefAuthor>Brachmann AO</RefAuthor> <RefAuthor>Kaiser M</RefAuthor> <RefAuthor>Bode HB</RefAuthor> <RefTitle>Bioactive derivatives of isopropylstilbene from mutasynthesis and chemical synthesis</RefTitle> <RefYear>2014</RefYear> <RefJournal>Chembiochem</RefJournal> <RefPage>2689-91</RefPage> <RefTotal>Kronenwerth M, Brachmann AO, Kaiser M, Bode HB. Bioactive derivatives of isopropylstilbene from mutasynthesis and chemical synthesis. Chembiochem. 2014 Dec;15(18):2689-91. DOI: 10.1002/cbic.201402447</RefTotal> <RefLink>http://dx.doi.org/10.1002/cbic.201402447</RefLink> </Reference> <Reference refNo="13"> <RefAuthor>Joyce SA</RefAuthor> <RefAuthor>Brachmann AO</RefAuthor> <RefAuthor>Glazer I</RefAuthor> <RefAuthor>Lango L</RefAuthor> <RefAuthor>Schwär G</RefAuthor> <RefAuthor>Clarke DJ</RefAuthor> <RefAuthor>Bode HB</RefAuthor> <RefTitle>Bacterial biosynthesis of a multipotent stilbene</RefTitle> <RefYear>2008</RefYear> <RefJournal>Angew Chem Int Ed Engl</RefJournal> <RefPage>1942-5</RefPage> <RefTotal>Joyce SA, Brachmann AO, Glazer I, Lango L, Schwär G, Clarke DJ, Bode HB. Bacterial biosynthesis of a multipotent stilbene. Angew Chem Int Ed Engl. 2008;47(10):1942-5. DOI: 10.1002/anie.200705148</RefTotal> <RefLink>http://dx.doi.org/10.1002/anie.200705148</RefLink> </Reference> <Reference refNo="14"> <RefAuthor>Buscató E</RefAuthor> <RefAuthor>Wisniewska JM</RefAuthor> <RefAuthor>Rödl CB</RefAuthor> <RefAuthor>Brüggerhoff A</RefAuthor> <RefAuthor>Kaiser A</RefAuthor> <RefAuthor>Rörsch F</RefAuthor> <RefAuthor>Kostewicz E</RefAuthor> <RefAuthor>Wurglics M</RefAuthor> <RefAuthor>Schubert-Zsilavecz M</RefAuthor> <RefAuthor>Grösch S</RefAuthor> <RefAuthor>Steinhilber D</RefAuthor> <RefAuthor>Hofmann B</RefAuthor> <RefAuthor>Proschak E</RefAuthor> <RefTitle>Structure-activity relationship and in vitro pharmacological evaluation of imidazo[1,2-a]pyridine-based inhibitors of 5-LO</RefTitle> <RefYear>2013</RefYear> <RefJournal>Future Med Chem</RefJournal> <RefPage>865-80</RefPage> <RefTotal>Buscató E, Wisniewska JM, Rödl CB, Brüggerhoff A, Kaiser A, Rörsch F, Kostewicz E, Wurglics M, Schubert-Zsilavecz M, Grösch S, Steinhilber D, Hofmann B, Proschak E. Structure-activity relationship and in vitro pharmacological evaluation of imidazo[1,2-a]pyridine-based inhibitors of 5-LO. Future Med Chem. 2013 May;5(8):865-80. DOI: 10.4155/fmc.13.72</RefTotal> <RefLink>http://dx.doi.org/10.4155/fmc.13.72</RefLink> </Reference> <Reference refNo="15"> <RefAuthor>Buscató E</RefAuthor> <RefAuthor>Blöcher R</RefAuthor> <RefAuthor>Lamers C</RefAuthor> <RefAuthor>Klingler FM</RefAuthor> <RefAuthor>Hahn S</RefAuthor> <RefAuthor>Steinhilber D</RefAuthor> <RefAuthor>Schubert-Zsilavecz M</RefAuthor> <RefAuthor>Proschak E</RefAuthor> <RefTitle>Design and synthesis of dual modulators of soluble epoxide hydrolase and peroxisome proliferator-activated receptors</RefTitle> <RefYear>2012</RefYear> <RefJournal>J Med Chem</RefJournal> <RefPage>10771-5</RefPage> <RefTotal>Buscató E, Blöcher R, Lamers C, Klingler FM, Hahn S, Steinhilber D, Schubert-Zsilavecz M, Proschak E. Design and synthesis of dual modulators of soluble epoxide hydrolase and peroxisome proliferator-activated receptors. J Med Chem. 2012 Dec;55(23):10771-5. DOI: 10.1021/jm301194c</RefTotal> <RefLink>http://dx.doi.org/10.1021/jm301194c</RefLink> </Reference> <Reference refNo="16"> <RefAuthor>Springer B</RefAuthor> <RefAuthor>Lucke K</RefAuthor> <RefAuthor>Calligaris-Maibach R</RefAuthor> <RefAuthor>Ritter C</RefAuthor> <RefAuthor>Böttger EC</RefAuthor> <RefTitle>Quantitative drug susceptibility testing of Mycobacterium tuberculosis by use of MGIT 960 and EpiCenter instrumentation</RefTitle> <RefYear>2009</RefYear> <RefJournal>J Clin Microbiol</RefJournal> <RefPage>1773-80</RefPage> <RefTotal>Springer B, Lucke K, Calligaris-Maibach R, Ritter C, Böttger EC. Quantitative drug susceptibility testing of Mycobacterium tuberculosis by use of MGIT 960 and EpiCenter instrumentation. J Clin Microbiol. 2009 Jun;47(6):1773-80. DOI: 10.1128/JCM.02501-08</RefTotal> <RefLink>http://dx.doi.org/10.1128/JCM.02501-08</RefLink> </Reference> <Reference refNo="17"> <RefAuthor>Michaelis M</RefAuthor> <RefAuthor>Matousek J</RefAuthor> <RefAuthor>Vogel JU</RefAuthor> <RefAuthor>Slavik T</RefAuthor> <RefAuthor>Langer K</RefAuthor> <RefAuthor>Cinatl J</RefAuthor> <RefAuthor>Kreuter J</RefAuthor> <RefAuthor>Schwabe D</RefAuthor> <RefAuthor>Cinatl J</RefAuthor> <RefTitle>Bovine seminal ribonuclease attached to nanoparticles made of polylactic acid kills leukemia and lymphoma cell lines in vitro</RefTitle> <RefYear>2000</RefYear> <RefJournal>Anticancer Drugs</RefJournal> <RefPage>369-76</RefPage> <RefTotal>Michaelis M, Matousek J, Vogel JU, Slavik T, Langer K, Cinatl J, Kreuter J, Schwabe D, Cinatl J. Bovine seminal ribonuclease attached to nanoparticles made of polylactic acid kills leukemia and lymphoma cell lines in vitro. Anticancer Drugs. 2000 Jun;11(5):369-76. DOI: 10.1097/00001813-200006000-00007</RefTotal> <RefLink>http://dx.doi.org/10.1097/00001813-200006000-00007</RefLink> </Reference> <Reference refNo="18"> <RefAuthor>Franzblau SG</RefAuthor> <RefAuthor>DeGroote MA</RefAuthor> <RefAuthor>Cho SH</RefAuthor> <RefAuthor>Andries K</RefAuthor> <RefAuthor>Nuermberger E</RefAuthor> <RefAuthor>Orme IM</RefAuthor> <RefAuthor>Mdluli K</RefAuthor> <RefAuthor>Angulo-Barturen I</RefAuthor> <RefAuthor>Dick T</RefAuthor> <RefAuthor>Dartois V</RefAuthor> <RefAuthor>Lenaerts AJ</RefAuthor> <RefTitle>Comprehensive analysis of methods used for the evaluation of compounds against Mycobacterium tuberculosis</RefTitle> <RefYear>2012</RefYear> <RefJournal>Tuberculosis (Edinb)</RefJournal> <RefPage>453-88</RefPage> <RefTotal>Franzblau SG, DeGroote MA, Cho SH, Andries K, Nuermberger E, Orme IM, Mdluli K, Angulo-Barturen I, Dick T, Dartois V, Lenaerts AJ. Comprehensive analysis of methods used for the evaluation of compounds against Mycobacterium tuberculosis. Tuberculosis (Edinb). 2012 Nov;92(6):453-88. DOI: 10.1016/j.tube.2012.07.003</RefTotal> <RefLink>http://dx.doi.org/10.1016/j.tube.2012.07.003</RefLink> </Reference> <Reference refNo="19"> <RefAuthor>Zhao L</RefAuthor> <RefAuthor>Chen X</RefAuthor> <RefAuthor>Cai L</RefAuthor> <RefAuthor>Zhang C</RefAuthor> <RefAuthor>Wang Q</RefAuthor> <RefAuthor>Jing S</RefAuthor> <RefAuthor>Chen G</RefAuthor> <RefAuthor>Li J</RefAuthor> <RefAuthor>Zhang J</RefAuthor> <RefAuthor>Fang Y</RefAuthor> <RefTitle>Randomized, double-blind, placebo-controlled, multiple-dose study of the safety, tolerability and pharmacokinetics of benvitimod, a candidate drug for the treatment of psoriasis</RefTitle> <RefYear>2014</RefYear> <RefJournal>J Clin Pharm Ther</RefJournal> <RefPage>418-23</RefPage> <RefTotal>Zhao L, Chen X, Cai L, Zhang C, Wang Q, Jing S, Chen G, Li J, Zhang J, Fang Y. Randomized, double-blind, placebo-controlled, multiple-dose study of the safety, tolerability and pharmacokinetics of benvitimod, a candidate drug for the treatment of psoriasis. J Clin Pharm Ther. 2014 Aug;39(4):418-23. DOI: 10.1111/jcpt.12158</RefTotal> <RefLink>http://dx.doi.org/10.1111/jcpt.12158</RefLink> </Reference> <Reference refNo="20"> <RefAuthor>Sirerol JA</RefAuthor> <RefAuthor>Rodríguez ML</RefAuthor> <RefAuthor>Mena S</RefAuthor> <RefAuthor>Asensi MA</RefAuthor> <RefAuthor>Estrela JM</RefAuthor> <RefAuthor>Ortega AL</RefAuthor> <RefTitle>Role of Natural Stilbenes in the Prevention of Cancer</RefTitle> <RefYear>2016</RefYear> <RefJournal>Oxid Med Cell Longev</RefJournal> <RefPage>3128951</RefPage> <RefTotal>Sirerol JA, Rodríguez ML, Mena S, Asensi MA, Estrela JM, Ortega AL. Role of Natural Stilbenes in the Prevention of Cancer. Oxid Med Cell Longev. 2016;2016:3128951. DOI: 10.1155/2016/3128951</RefTotal> <RefLink>http://dx.doi.org/10.1155/2016/3128951</RefLink> </Reference> </References> <Media> <Tables> <Table format="png"> <MediaNo>1</MediaNo> <MediaID>1</MediaID> <Caption><Pgraph><Mark1>Table 1: Anti-tubercular activity of stilbenes </Mark1><LineBreak></LineBreak>MIC of substances against <Mark2>M. tuberculosis</Mark2> ATCC 27294 as well as MDR and XDR <Mark2>M. tuberculosis</Mark2> clinical isolates </Pgraph></Caption> </Table> <Table format="png"> <MediaNo>2</MediaNo> <MediaID>2</MediaID> <Caption><Pgraph><Mark1>Table 2: Toxicity of stilbenes</Mark1><LineBreak></LineBreak>IC<Subscript>50</Subscript> of selected substances</Pgraph></Caption> </Table> <NoOfTables>2</NoOfTables> </Tables> <Figures> <Figure format="png" height="366" width="592"> <MediaNo>1</MediaNo> <MediaID>1</MediaID> <Caption><Pgraph><Mark1>Figure 1: Natural stilbene and its synthetic derivatives evaluated in this study</Mark1><LineBreak></LineBreak>a) Natural stilbene, isopropylstilbene (HB64); b) (E)-styryl-1H-benzo[d]imidazoles with DB53 (R=F), DB55 (R=CH3), DB56 (R=Cl); c) N-((3s, 5s, 7s)-adamantam-1-yl)-6-chloro-2-(4-morpholinophenyl)imidazo[1, 2-a]pyridine-3-amine (EB251); d) 1-(3-((3-cyclohexylureido)methyl)benzyl)-1H-pyrrole-2-carboxylate (RB57)</Pgraph></Caption> </Figure> <NoOfPictures>1</NoOfPictures> </Figures> <InlineFigures> <NoOfPictures>0</NoOfPictures> </InlineFigures> <Attachments> <NoOfAttachments>0</NoOfAttachments> </Attachments> </Media> </OrigData> </GmsArticle>