scholarly journals Proteomic Signature of Fatty Acid Biosynthesis Inhibition Available for In Vivo Mechanism-of-Action Studies

2011 ◽  
Vol 55 (6) ◽  
pp. 2590-2596 ◽  
Author(s):  
Michaela Wenzel ◽  
Malay Patra ◽  
Dirk Albrecht ◽  
David Y.-K. Chen ◽  
K. C. Nicolaou ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Structure Activity Relationship ◽  
Mechanisms Of Action ◽  
Activity Relationship ◽  
Antibacterial Drug ◽  
Acid Biosynthesis ◽  
Content Type ◽  
Structure Activity

ABSTRACTFatty acid biosynthesis is a promising novel antibiotic target. Two inhibitors of fatty acid biosynthesis, platencin and platensimycin, were recently discovered and their molecular targets identified. Numerous structure-activity relationship studies for both platencin and platensimycin are currently being undertaken. We established a proteomic signature for fatty acid biosynthesis inhibition inBacillus subtilisusing platencin, platensimycin, cerulenin, and triclosan. The induced proteins, FabHA, FabHB, FabF, FabI, PlsX, and PanB, are enzymes involved in fatty acid biosynthesis and thus linked directly to the target pathway. The proteomic signature can now be used to assess thein vivomechanisms of action of compounds derived from structure-activity relationship programs, as demonstrated for the platensimycin-inspired chromium bioorganometallic PM47. It will further serve as a reference signature for structurally novel natural and synthetic antimicrobial compounds with unknown mechanisms of action. In summary, we described a proteomic signature inB. subtilisconsisting of six upregulated proteins that is diagnostic of fatty acid biosynthesis inhibition and thus can be applied to advance antibacterial drug discovery programs.

scholarly journals Identification and Characterization of Inhibitors of Bacterial Enoyl-Acyl Carrier Protein Reductase

2004 ◽  
Vol 48 (5) ◽  
pp. 1541-1547 ◽  
Author(s):  
Losee L. Ling ◽  
Jun Xian ◽  
Syed Ali ◽  
Bolin Geng ◽  
Jun Fan ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Acyl Carrier Protein ◽  
Bacterial Species ◽  
Antibacterial Drug ◽  
Initial Structure ◽  
Carrier Protein ◽  
Acid Biosynthesis ◽  
Structure Activity Relationships ◽  
Structure Activity

ABSTRACT Bacterial enoyl-acyl carrier protein reductase (ENR) catalyzes an essential step in fatty acid biosynthesis. ENR is an attractive target for narrow-spectrum antibacterial drug discovery because of its essential role in metabolism and its sequence conservation across many bacterial species. In addition, the bacterial ENR sequence and structural organization are distinctly different from those of mammalian fatty acid biosynthesis enzymes. High-throughput screening to identify inhibitors of Escherichia coli ENR yielded four structurally distinct classes of hits. Several members of one of these, the 2-(alkylthio)-4,6-diphenylpyridine-3-carbonitriles (“thiopyridines”), inhibited both purified ENR (50% inhibitory concentration [IC50] = 3 to 25 μM) and the growth of Staphylococcus aureus and Bacillus subtilis (MIC = 1 to 64 μg/ml). The effect on cell growth is due in part to inhibition of fatty acid biosynthesis as judged by inhibition of incorporation of [14C]acetate into fatty acids and by the increased sensitivity of cells that underexpress an ENR-encoding gene (four- to eightfold MIC shift). Synthesis of a variety of compounds in this chemical series revealed a correlation between IC50 and MIC, and the results provided initial structure-activity relationships. Preliminary structure-activity relationships, potency on purified ENR, and activity on bacterial cells indicate that members of the thiopyridine chemical series are effective fatty acid biosynthesis inhibitors suitable for further antibacterial development.

scholarly journals In Vivo Roles of Fatty Acid Biosynthesis Enzymes in Biosynthesis of Biotin and α-Lipoic Acid in Corynebacterium glutamicum

2017 ◽  
Vol 83 (19) ◽  
Author(s):  
Masato Ikeda ◽  
Takashi Nagashima ◽  
Eri Nakamura ◽  
Ryosuke Kato ◽  
Masakazu Ohshita ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipoic Acid ◽  
Fatty Acid Synthase ◽  
Fatty Acid Biosynthesis ◽  
Physiological Role ◽  
Type I ◽  
Acid Biosynthesis ◽  
Content Type ◽  
Fas Ii

ABSTRACT For fatty acid biosynthesis, Corynebacterium glutamicum uses two type I fatty acid synthases (FAS-I), FasA and FasB, in addition to acetyl-coenzyme A (CoA) carboxylase (ACC) consisting of AccBC, AccD1, and AccE. The in vivo roles of the enzymes in supplying precursors for biotin and α-lipoic acid remain unclear. Here, we report genetic evidence demonstrating that the biosynthesis of these cofactors is linked to fatty acid biosynthesis through the FAS-I pathway. For this study, we used wild-type C. glutamicum and its derived biotin vitamer producer BFI-5, which was engineered to express Escherichia coli bioBF and Bacillus subtilis bioI. Disruption of either fasA or fasB in strain BFI-5 led to decreased production of biotin vitamers, whereas its amplification contributed to increased production, with a larger impact of fasA in both cases. Double disruptions of fasA and fasB resulted in no biotin vitamer production. The acc genes showed a positive effect on production when amplified simultaneously. Augmented fatty acid biosynthesis was also reflected in pimelic acid production when carbon flow was blocked at the BioF reaction. These results indicate that carbon flow down the FAS-I pathway is destined for channeling into the biotin biosynthesis pathway, and that FasA in particular has a significant impact on precursor supply. In contrast, fasB disruption resulted in auxotrophy for lipoic acid or its precursor octanoic acid in both wild-type and BFI-5 strains. The phenotypes were fully complemented by plasmid-mediated expression of fasB but not fasA. These results reveal that FasB plays a specific physiological role in lipoic acid biosynthesis in C. glutamicum. IMPORTANCE For the de novo biosynthesis of fatty acids, C. glutamicum exceptionally uses a eukaryotic multifunctional type I fatty acid synthase (FAS-I) system comprising FasA and FasB, in contrast to most bacteria, such as E. coli and B. subtilis, which use an individual nonaggregating type II fatty acid synthase (FAS-II) system. In this study, we reported genetic evidence demonstrating that the FAS-I system is the source of the biotin precursor in vivo in the engineered biotin-prototrophic C. glutamicum strain. This study also uncovered the important physiological role of FasB in lipoic acid biosynthesis. Here, we present an FAS-I enzyme that functions in supplying the lipoic acid precursor, although its biosynthesis has been believed to exclusively depend on FAS-II in organisms. The findings obtained here provide new insights into the metabolic engineering of this industrially important microorganism to produce these compounds effectively.

scholarly journals Optimization of Pyrrolamides as Mycobacterial GyrB ATPase Inhibitors: Structure-Activity Relationship andIn VivoEfficacy in a Mouse Model of Tuberculosis

2013 ◽  
Vol 58 (1) ◽  
pp. 61-70 ◽  
Author(s):  
Shahul Hameed P ◽  
Suresh Solapure ◽  
Kakoli Mukherjee ◽  
Vrinda Nandi ◽  
David Waterson ◽  
...  
Keyword(s):  
Mouse Model ◽  
Dna Gyrase ◽  
Structure Activity Relationship ◽  
Activity Relationship ◽  
Drug Resistant ◽  
Binding Interactions ◽  
Content Type ◽  
Log Reduction ◽  
Structure Activity

ABSTRACTMoxifloxacin has shown excellent activity against drug-sensitive as well as drug-resistant tuberculosis (TB), thus confirming DNA gyrase as a clinically validated target for discovering novel anti-TB agents. We have identified novel inhibitors in the pyrrolamide class which killMycobacterium tuberculosisthrough inhibition of ATPase activity catalyzed by the GyrB domain of DNA gyrase. A homology model of theM. tuberculosisH37Rv GyrB domain was used for deciphering the structure-activity relationship and binding interactions of inhibitors with mycobacterial GyrB enzyme. Proposed binding interactions were later confirmed through cocrystal structure studies with theMycobacterium smegmatisGyrB ATPase domain. The most potent compound in this series inhibited supercoiling activity of DNA gyrase with a 50% inhibitory concentration (IC50) of <5 nM, an MIC of 0.03 μg/ml againstM. tuberculosisH37Rv, and an MIC90of <0.25 μg/ml against 99 drug-resistant clinical isolates ofM. tuberculosis. The frequency of isolating spontaneous resistant mutants was ∼10−6to 10−8, and the point mutation mapped to theM. tuberculosisGyrB domain (Ser208 Ala), thus confirming its mode of action. The best compound tested forin vivoefficacy in the mouse model showed a 1.1-log reduction in lung CFU in the acute model and a 0.7-log reduction in the chronic model. This class of GyrB inhibitors could be developed as novel anti-TB agents.

scholarly journals Synthesis, Anticancer Activity, and Preliminary Pharmacokinetic Evaluation of 4,4-Disubstituted Curcuminoid 2,2-bis(Hydroxymethyl)Propionate Derivatives

Molecules ◽  
2020 ◽  
Vol 25 (3) ◽  
pp. 479 ◽  
Author(s):  
Der-Yen Lee ◽  
Yu-Chi Hou ◽  
Jai-Sing Yang ◽  
Hui-Yi Lin ◽  
Tsu-Yuan Chang ◽  
...  
Keyword(s):  
Anticancer Activity ◽  
Functional Groups ◽  
Hydrolysis Product ◽  
Structure Activity Relationship ◽  
Ester Hydrolysis ◽  
Activity Relationship ◽  
Structure Activity ◽  
Hct 116 ◽  
Pharmacokinetic Evaluation

Compound 1 is a curcumin di-O-2,2-bis(hydroxymethyl)propionate that shows significant in vitro and in vivo inhibitory activity against MDA-MB-231 cells with eight to ten-fold higher potency than curcumin. Here, we modified the α-position (C-4 position) of the central 1,3-diketone moiety of 1 with polar or nonpolar functional groups to afford a series of 4,4-disubstituted curcuminoid 2,2-bis(hydroxymethyl)propionate derivatives and evaluated their anticancer activities. A clear structure–activity relationship of compound 1 derivatives focusing on the functional groups at the C-4 position was established based on their anti-proliferative effects against the MDA-MB-231 and HCT-116 cell lines. Compounds 2–6 are 4,4-dimethylated, 4,4-diethylated, 4,4-dibenzylated, 4,4-dipropargylated and 4,4-diallylated compound 1, respectively. Compounds 2m–6m, the ester hydrolysis products of compounds 2–6, respectively, were synthesized and assessed for anticancer activity. Among all compound 1 derivatives, compound 2 emerged as a potential chemotherapeutic agent for colon cancer due to the promising in vivo anti-proliferative activities of 2 (IC50 = 3.10 ± 0.29 μM) and its ester hydrolysis product 2m (IC50 = 2.17 ± 0.16 μM) against HCT-116. The preliminary pharmacokinetic evaluation of 2 implied that 2 and 2m are main contributors to the in vivo efficacy. Compound 2 was further evaluated in an animal study using HCT-116 colon tumor xenograft bearing nude mice. The results revealed a dose-dependent efficacy that led to tumor volume reductions of 27%, 45%, and 60% at 50, 100, and 150 mg/kg doses, respectively. The established structure–activity relationship and pharmacokinetic outcomes of 2 is the guidance for future development of 4,4-disubstituted curcuminoid 2,2-bis(hydroxymethyl)- propionate derivatives as anticancer drug candidates.

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