|
Binding affinity against Synechocystis strain PCC6803 DXP reductoisomerase
|
Synechocystis sp. PCC 6803
|
21.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Substrate analogs for the investigation of deoxyxylulose 5-phosphate reductoisomerase inhibition: synthesis and evaluation.
Year : 2004
Volume : 14
Issue : 21
First Page : 5309
Last Page : 5312
Authors : Phaosiri C, Proteau PJ.
Abstract : Deoxyxylulose 5-phosphate (DXP) analogs were synthesized and evaluated as alternative substrates and inhibitors of recombinant Synechocystis PCC6803 DXP reductoisomerase (DXR; EC 1.1.1.267). Five of the compounds tested (1,2-dideoxy-D-threo-3-hexulose 6-phosphate, 1-deoxy-l-ribulose 5-phosphate, 2S,3R-dihydroxybutyramide 4-phosphate, 4S-hydroxypentan-2-one 5-phosphate, and 3S-hydroxypentan-2-one 5-phosphate) acted as relatively weak competitive inhibitors when compared to fosmidomycin. A sixth compound, 3R,4S-dihydroxy-5-oxohexylphosphonic acid, served as an alternate substrate, as has recently been reported for the same compound with Escherichia coli DXR.
|
Inhibitory concentration against DOXP reductoisomerase
|
Escherichia coli
|
19.95
nM
|
|
Journal : J. Med. Chem.
Title : AFMoC enhances predictivity of 3D QSAR: a case study with DOXP-reductoisomerase.
Year : 2005
Volume : 48
Issue : 10
First Page : 3547
Last Page : 3563
Authors : Silber K, Heidler P, Kurz T, Klebe G.
Abstract : We present structure-activity relationships for 43 inhibitors of 1-deoxyxylulose-5-phosphate (DOXP)-reductoisomerase, derived from protein-based docking, ligand-based 3D QSAR, and a combination of both approaches as realized by AFMoC (adaptation of fields for molecular comparison). DOXP-reductoisomerase (DXR) is a key enzyme of the non-mevalonate pathway for isoprenoid building blocks. This target has been characterized as having potential in the treatment of malaria with fosmidomycin, an established DXR inhibitor, presently in clinical trials. As part of an effort to optimize the properties of fosmidomycin, analogues have been synthesized and tested to gain further insights into the primary determinants of structural affinity. These data have been used to create a predictive model for DXR inhibition applying data taken from several DXR X-ray structures. These structures still leave the active fosmidomycin conformation and detailed reaction mechanism undetermined. This together with the small inhibitor data set provides a major challenge for presently available docking programs and 3D QSAR tools. To overcome these difficulties we have applied the AFMoC protocol. AFMoC makes more efficient use of available modeling data by tailoring DrugScore knowledge-based potentials specifically toward a given protein using inhibitor potency data. While 3D QSAR methods achieved valid models which lack predictivity, AFMoC was found to provide superior performance, based both on cross-validation runs as well as for inhibitors not considered in the training set. In particular, AFMoC's ability to gradually transform between generally applicable unadapted interaction fields to case specifically adapted ones proved to be of major importance. Using 50% tailored fields was found to permit the precise prediction of binding affinities for related ligands without losing the capability to estimate the affinities of structurally distinct inhibitors.
|
Inhibition of recombinant Escherichia coli DXR
|
Escherichia coli
|
48.0
nM
|
|
Journal : J. Med. Chem.
Title : Synthesis and biological evaluation of cyclopropyl analogues of fosmidomycin as potent Plasmodium falciparum growth inhibitors.
Year : 2006
Volume : 49
Issue : 8
First Page : 2656
Last Page : 2660
Authors : Devreux V, Wiesner J, Goeman JL, Van der Eycken J, Jomaa H, Van Calenbergh S.
Abstract : A series of fosmidomycin analogues featuring restricted conformational mobility has been synthesized and evaluated as inhibitors of 1-deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase and as growth inhibitors of P. falciparum. The enantiomerically pure trans-cyclopropyl N-acetyl analogue 3b showed comparable inhibitory activity as fosmidomycin toward E. coli DOXP reductoisomerase and proved equally active when tested in vitro for P. falciparum growth inhibition. Conversely, the alpha-phenyl cis-cyclopropyl analogue 4 showed virtually no inhibition of the enzyme.
|
Inhibition of Plasmodium falciparum Dd2
|
Plasmodium falciparum
|
480.0
nM
|
|
Journal : J. Med. Chem.
Title : Synthesis and biological evaluation of cyclopropyl analogues of fosmidomycin as potent Plasmodium falciparum growth inhibitors.
Year : 2006
Volume : 49
Issue : 8
First Page : 2656
Last Page : 2660
Authors : Devreux V, Wiesner J, Goeman JL, Van der Eycken J, Jomaa H, Van Calenbergh S.
Abstract : A series of fosmidomycin analogues featuring restricted conformational mobility has been synthesized and evaluated as inhibitors of 1-deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase and as growth inhibitors of P. falciparum. The enantiomerically pure trans-cyclopropyl N-acetyl analogue 3b showed comparable inhibitory activity as fosmidomycin toward E. coli DOXP reductoisomerase and proved equally active when tested in vitro for P. falciparum growth inhibition. Conversely, the alpha-phenyl cis-cyclopropyl analogue 4 showed virtually no inhibition of the enzyme.
|
Inhibition of Plasmodium falciparum 3D7
|
Plasmodium falciparum
|
400.0
nM
|
|
Journal : J. Med. Chem.
Title : Synthesis and biological evaluation of cyclopropyl analogues of fosmidomycin as potent Plasmodium falciparum growth inhibitors.
Year : 2006
Volume : 49
Issue : 8
First Page : 2656
Last Page : 2660
Authors : Devreux V, Wiesner J, Goeman JL, Van der Eycken J, Jomaa H, Van Calenbergh S.
Abstract : A series of fosmidomycin analogues featuring restricted conformational mobility has been synthesized and evaluated as inhibitors of 1-deoxy-D-xylulose 5-phosphate (DOXP) reductoisomerase and as growth inhibitors of P. falciparum. The enantiomerically pure trans-cyclopropyl N-acetyl analogue 3b showed comparable inhibitory activity as fosmidomycin toward E. coli DOXP reductoisomerase and proved equally active when tested in vitro for P. falciparum growth inhibition. Conversely, the alpha-phenyl cis-cyclopropyl analogue 4 showed virtually no inhibition of the enzyme.
|
Inhibition of recombinant Escherichia coli DXR
|
Escherichia coli
|
30.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Synthesis of alpha-substituted fosmidomycin analogues as highly potent Plasmodium falciparum growth inhibitors.
Year : 2006
Volume : 16
Issue : 7
First Page : 1888
Last Page : 1891
Authors : Haemers T, Wiesner J, Van Poecke S, Goeman J, Henschker D, Beck E, Jomaa H, Van Calenbergh S.
Abstract : In view of the promising antimalarial activity of fosmidomycin or its N-acetyl homologue FR900098, the objective of this work was to investigate the influence of aromatic substituents in the alpha-position of the phosphonate moiety. The envisaged analogues were prepared using a linear route involving a 3-aryl-3-phosphoryl propanal intermediate. The activities of all compounds were evaluated on Eschericia coli 1-deoxy-d-xylulose 5-phosphate reductoisomerase and against two Plasmodium falciparum strains. Compared with fosmidomycin, several analogues displayed enhanced activity towards the P. falciparum strains. Compound 1e with a 3,4-dichlorophenyl substitution in the alpha-position of fosmidomycin emerged as the most potent analogue of this series. It is approximately three times more potent in inhibiting the growth of P. falciparum than FR900098, the most potent representative of this class reported so far.
|
Antimalarial activity against Plasmodium falciparum Dd2
|
Plasmodium falciparum
|
360.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Synthesis of alpha-substituted fosmidomycin analogues as highly potent Plasmodium falciparum growth inhibitors.
Year : 2006
Volume : 16
Issue : 7
First Page : 1888
Last Page : 1891
Authors : Haemers T, Wiesner J, Van Poecke S, Goeman J, Henschker D, Beck E, Jomaa H, Van Calenbergh S.
Abstract : In view of the promising antimalarial activity of fosmidomycin or its N-acetyl homologue FR900098, the objective of this work was to investigate the influence of aromatic substituents in the alpha-position of the phosphonate moiety. The envisaged analogues were prepared using a linear route involving a 3-aryl-3-phosphoryl propanal intermediate. The activities of all compounds were evaluated on Eschericia coli 1-deoxy-d-xylulose 5-phosphate reductoisomerase and against two Plasmodium falciparum strains. Compared with fosmidomycin, several analogues displayed enhanced activity towards the P. falciparum strains. Compound 1e with a 3,4-dichlorophenyl substitution in the alpha-position of fosmidomycin emerged as the most potent analogue of this series. It is approximately three times more potent in inhibiting the growth of P. falciparum than FR900098, the most potent representative of this class reported so far.
|
Inhibition of Trypanosoma brucei FPPS expressed in Escherichia coli
|
Trypanosoma brucei
|
200.0
nM
|
|
Journal : J. Med. Chem.
Title : Isoprenoid biosynthesis as a drug target: bisphosphonate inhibition of Escherichia coli K12 growth and synergistic effects of fosmidomycin.
Year : 2006
Volume : 49
Issue : 25
First Page : 7331
Last Page : 7341
Authors : Leon A, Liu L, Yang Y, Hudock MP, Hall P, Yin F, Studer D, Puan KJ, Morita CT, Oldfield E.
Abstract : We screened a library of 117 bisphosphonates for antibacterial activity against Escherichia coli. The most potent growth inhibitors where N-[methyl(4-phenylalkyl)]-3-aminopropyl-1-hydroxy-1,1-bisphosphonates, known potent bone resorption inhibitors, and there was a generally good correlation between cell growth inhibition and E. coli farnesyl diphosphate synthase (FPPS) inhibition. However, some potent FPPS inhibitors had no activity in cell growth inhibition, and based on the result of Catalyst pharmacophore modeling, this could be attributed to the requirement of a large hydrophobic feature for cellular activity (due most likely to transport). The activity of the most potent compound, N-[methyl(4-phenylbutyl)]-3-aminopropyl-1-hydroxy-1,1-bisphosphonate (13), was strongly potentiated by the drug fosmidomycin. The transcription profiles for 13 or fosmidomycin alone were different from those found with carbenicillin or ciprofloxacin alone, but there were many similarities between the combination (13-fosmidomycin) and carbenicillin or ciprofloxacin, reflecting the more potent bactericidal activity of the drug combination on bacterial growth.
|
Antibacterial activity against Escherichia coli K12 by broth microdilution method
|
Escherichia coli
|
220.0
nM
|
|
Journal : J. Med. Chem.
Title : Isoprenoid biosynthesis as a drug target: bisphosphonate inhibition of Escherichia coli K12 growth and synergistic effects of fosmidomycin.
Year : 2006
Volume : 49
Issue : 25
First Page : 7331
Last Page : 7341
Authors : Leon A, Liu L, Yang Y, Hudock MP, Hall P, Yin F, Studer D, Puan KJ, Morita CT, Oldfield E.
Abstract : We screened a library of 117 bisphosphonates for antibacterial activity against Escherichia coli. The most potent growth inhibitors where N-[methyl(4-phenylalkyl)]-3-aminopropyl-1-hydroxy-1,1-bisphosphonates, known potent bone resorption inhibitors, and there was a generally good correlation between cell growth inhibition and E. coli farnesyl diphosphate synthase (FPPS) inhibition. However, some potent FPPS inhibitors had no activity in cell growth inhibition, and based on the result of Catalyst pharmacophore modeling, this could be attributed to the requirement of a large hydrophobic feature for cellular activity (due most likely to transport). The activity of the most potent compound, N-[methyl(4-phenylbutyl)]-3-aminopropyl-1-hydroxy-1,1-bisphosphonate (13), was strongly potentiated by the drug fosmidomycin. The transcription profiles for 13 or fosmidomycin alone were different from those found with carbenicillin or ciprofloxacin alone, but there were many similarities between the combination (13-fosmidomycin) and carbenicillin or ciprofloxacin, reflecting the more potent bactericidal activity of the drug combination on bacterial growth.
|
Inhibition of Escherichia coli DXR
|
Escherichia coli
|
34.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Synthesis and evaluation of alpha,beta-unsaturated alpha-aryl-substituted fosmidomycin analogues as DXR inhibitors.
Year : 2007
Volume : 17
Issue : 17
First Page : 4920
Last Page : 4923
Authors : Devreux V, Wiesner J, Jomaa H, Van der Eycken J, Van Calenbergh S.
Abstract : Fosmidomycin, which acts through inhibition of 1-deoxy-D-xylulose phosphate reductoisomerase (DXR) in the non-mevalonate pathway, represents a valuable recent addition to the armamentarium against uncomplicated malaria. In this paper, we describe the synthesis and biological evaluation of E- and Z-alpha,beta-unsaturated alpha-aryl-substituted analogues of FR900098, a fosmidomycin congener, utilizing a Stille or a Suzuki coupling to introduce the aryl group. In contrast with our expectations based on the promising activity earlier observed for several alpha-substituted fosmidomycin analogues, all synthesized analogues exhibited much lower binding affinity for DXR than fosmidomycin.
|
Inhibition of Escherichia coli DXR
|
Escherichia coli
|
54.0
nM
|
|
Journal : Bioorg. Med. Chem.
Title : Synthesis of beta- and gamma-oxa isosteres of fosmidomycin and FR900098 as antimalarial candidates.
Year : 2008
Volume : 16
Issue : 6
First Page : 3361
Last Page : 3371
Authors : Haemers T, Wiesner J, Giessmann D, Verbrugghen T, Hillaert U, Ortmann R, Jomaa H, Link A, Schlitzer M, Van Calenbergh S.
Abstract : To expand the structure-activity relationships of fosmidomycin and FR900098, two potent antimalarials interfering with the MEP-pathway, we decided to replace a methylene group in beta-position of the phosphonate moiety of these leads by an oxygen atom. beta-oxa-FR900098 (11) proved equally active as the parent compound. When applied to 4-[hydroxyl(methyl)amino]-4-oxobutyl phosphonic acid, featuring a hydroxamate instead of the retrohydroxamate moiety, a beta-oxa modification yielded a derivative (13) with superior activity against the Plasmodium falciparum 3D7 strain than fosmidomycin, while a gamma-oxa modification resulted in less active derivatives. A bis(pivaloyloxymethyl)ester of phosphonate 13 proved twice as active in inhibiting cultured parasites as a similar prodrug of FR900098.
|
Inhibition of Escherichia coli DXR by spectrophotometric method
|
Escherichia coli
|
30.0
nM
|
|
Journal : J. Biol. Chem.
Title : Structures of Mycobacterium tuberculosis 1-deoxy-D-xylulose-5-phosphate reductoisomerase provide new insights into catalysis.
Year : 2007
Volume : 282
Issue : 27
First Page : 19905
Last Page : 19916
Authors : Henriksson LM, Unge T, Carlsson J, Aqvist J, Mowbray SL, Jones TA.
Abstract : Isopentenyl diphosphate is the precursor of various isoprenoids that are essential to all living organisms. It is produced by the mevalonate pathway in humans but by an alternate route in plants, protozoa, and many bacteria. 1-deoxy-D-xylulose-5-phosphate reductoisomerase catalyzes the second step of this non-mevalonate pathway, which involves an NADPH-dependent rearrangement and reduction of 1-deoxy-D-xylulose 5-phosphate to form 2-C-methyl-D-erythritol 4-phosphate. The use of different pathways, combined with the reported essentiality of the enzyme makes the reductoisomerase a highly promising target for drug design. Here we present several high resolution structures of the Mycobacterium tuberculosis 1-deoxy-D-xylulose-5-phosphate reductoisomerase, representing both wild type and mutant enzyme in various complexes with Mn(2+), NADPH, and the known inhibitor fosmidomycin. The asymmetric unit corresponds to the biological homodimer. Although crystal contacts stabilize an open active site in the B molecule, the A molecule displays a closed conformation, with some differences depending on the ligands bound. An inhibition study with fosmidomycin resulted in an estimated IC(50) value of 80 nm. The double mutant enzyme (D151N/E222Q) has lost its ability to bind the metal and, thereby, also its activity. Our structural information complemented with molecular dynamics simulations and free energy calculations provides the framework for the design of new inhibitors and gives new insights into the reaction mechanism. The conformation of fosmidomycin bound to the metal ion is different from that reported in a previously published structure and indicates that a rearrangement of the intermediate is not required during catalysis.
|
Inhibition of Mycobacterium tuberculosis DXR assessed as NADPH-dependent conversion of 1-deoxy-D-xylulose 5-phosphate to 2-C-methyl-D-erythritol 4-phosphate by spectrophotometric method
|
Mycobacterium tuberculosis
|
80.0
nM
|
|
Journal : J. Biol. Chem.
Title : Structures of Mycobacterium tuberculosis 1-deoxy-D-xylulose-5-phosphate reductoisomerase provide new insights into catalysis.
Year : 2007
Volume : 282
Issue : 27
First Page : 19905
Last Page : 19916
Authors : Henriksson LM, Unge T, Carlsson J, Aqvist J, Mowbray SL, Jones TA.
Abstract : Isopentenyl diphosphate is the precursor of various isoprenoids that are essential to all living organisms. It is produced by the mevalonate pathway in humans but by an alternate route in plants, protozoa, and many bacteria. 1-deoxy-D-xylulose-5-phosphate reductoisomerase catalyzes the second step of this non-mevalonate pathway, which involves an NADPH-dependent rearrangement and reduction of 1-deoxy-D-xylulose 5-phosphate to form 2-C-methyl-D-erythritol 4-phosphate. The use of different pathways, combined with the reported essentiality of the enzyme makes the reductoisomerase a highly promising target for drug design. Here we present several high resolution structures of the Mycobacterium tuberculosis 1-deoxy-D-xylulose-5-phosphate reductoisomerase, representing both wild type and mutant enzyme in various complexes with Mn(2+), NADPH, and the known inhibitor fosmidomycin. The asymmetric unit corresponds to the biological homodimer. Although crystal contacts stabilize an open active site in the B molecule, the A molecule displays a closed conformation, with some differences depending on the ligands bound. An inhibition study with fosmidomycin resulted in an estimated IC(50) value of 80 nm. The double mutant enzyme (D151N/E222Q) has lost its ability to bind the metal and, thereby, also its activity. Our structural information complemented with molecular dynamics simulations and free energy calculations provides the framework for the design of new inhibitors and gives new insights into the reaction mechanism. The conformation of fosmidomycin bound to the metal ion is different from that reported in a previously published structure and indicates that a rearrangement of the intermediate is not required during catalysis.
|
Inhibition of Escherichia coli M15 DXR
|
Escherichia coli
|
80.0
nM
|
|
Journal : J. Med. Chem.
Title : Coordination chemistry based approach to lipophilic inhibitors of 1-deoxy-D-xylulose-5-phosphate reductoisomerase.
Year : 2009
Volume : 52
Issue : 21
First Page : 6539
Last Page : 6542
Authors : Deng L, Sundriyal S, Rubio V, Shi ZZ, Song Y.
Abstract : 1-Deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) in the non-mevalonate pathway found in most bacteria is a validated anti-infective drug target. Fosmidomycin, a potent DXR inhibitor, is active against Gram-negative bacteria. A coordination chemistry and structure based approach was used to discover a novel, lipophilic DXR inhibitor with an IC(50) of 1.4 microM. It exhibited a broad spectrum of activity against Gram-negative and -positive bacteria with minimal inhibition concentrations of 20-100 microM (or 3.7-19 microg/mL).
|
Antiplasmodial activity against Plasmodium berghei ANKA infected in mice (Mus musculus) assessed as suppression of parasitaemia at 50 mg/kg, intraperitoneal for 5 consecutive days measured on day 4 post parasitic infection
|
Plasmodium berghei str. ANKA
|
82.0
%
|
|
Journal : J. Med. Chem.
Title : Synthesis and evaluation of alpha-halogenated analogues of 3-(acetylhydroxyamino)propylphosphonic acid (FR900098) as antimalarials.
Year : 2010
Volume : 53
Issue : 14
First Page : 5342
Last Page : 5346
Authors : Verbrugghen T, Cos P, Maes L, Van Calenbergh S.
Abstract : Three alpha-halogenated analogues of 3-(acetylhydroxyamino)propylphosphonic acid (FR900098) have been synthesized from diethyl but-3-enylphosphonate using a previously described method for the alpha-halogenation of alkylphosphonates. These analogues were evaluated for antimalarial potential in vitro against Plasmodium falciparum and in vivo in the P. berghei mouse model. All three analogues showed higher in vitro and/or in vivo potency than the reference compounds.
|
Inhibition of Escherichia coli DXR
|
Escherichia coli
|
30.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Structures of 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase/Lipophilic Phosphonate Complexes.
Year : 2011
Volume : 2
Issue : 2
First Page : 165
Last Page : 170
Authors : Deng L, Endo K, Kato M, Cheng G, Yajima S, Song Y.
Abstract : Fosmidomycin, a potent inhibitor of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), has antibacterial and antimalaria activity. Due to its poor pharmacokinetics, more lipophilic DXR inhibitors are needed. However, the hydrophobic binding site(s) in DXR remains elusive. Here, pyridine/quinoline containing phosphonates are identified to be DXR inhibitors with IC(50) values as low as 840 nM. We also report three DXR:inhibitor structures, revealing a novel binding mode. The indole group of Trp211 is found to move ~4.6 Å to open up a mainly hydrophobic pocket, where the pyridine/quinoline rings of the inhibitors are located and have strong π-π stacking/charge-transfer interactions with the indole. Docking studies demonstrate our structures could be used to predict the binding modes of other lipophilic DXR inhibitors. Overall, this work shows an important role of Trp211 in inhibitor recognition and provides a structural basis for future drug design and development.
|
Inhibition of Escherichia coli DXR assessed as conversion of NADPH to NADP at 0.3 uM by spectrophotometry
|
Escherichia coli
|
99.3
%
|
|
Journal : Bioorg. Med. Chem.
Title : Synthesis and evaluation of phosphonated N-heteroarylcarboxamides as DOXP-reductoisomerase (DXR) inhibitors.
Year : 2011
Volume : 19
Issue : 3
First Page : 1321
Last Page : 1327
Authors : Bodill T, Conibear AC, Blatch GL, Lobb KA, Kaye PT.
Abstract : The diethyl esters and disodium salts of a range of heteroarylcarbamoylphosphonic acids have been prepared and evaluated as analogues of the highly active DOXP-reductoisomerase (DXR) inhibitor, fosmidomycin. Computer-simulated docking studies, Saturation Transfer Difference (STD) NMR analysis and enzyme inhibition assays have been used to explore enzyme-binding and -inhibition potential, while in silico analysis of the DXR active site has highlighted the importance of including a well-parameterised metal co-factor in docking studies and has revealed the availability of an additional binding pocket to guide future drug design.
|
Inhibition of Escherichia coli recombinant DXR using DXP as substrate and MgCl2 as cofactor preincubated for 10 mins
|
Escherichia coli
|
26.92
nM
|
|
Inhibition of Escherichia coli recombinant DXR using DXP as substrate and MgCl2 as cofactor preincubated for 10 mins
|
Escherichia coli
|
27.0
nM
|
|
Journal : J. Med. Chem.
Title : Inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase by lipophilic phosphonates: SAR, QSAR, and crystallographic studies.
Year : 2011
Volume : 54
Issue : 13
First Page : 4721
Last Page : 4734
Authors : Deng L, Diao J, Chen P, Pujari V, Yao Y, Cheng G, Crick DC, Prasad BV, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) is a novel target for developing new antibacterial (including antituberculosis) and antimalaria drugs. Forty-one lipophilic phosphonates, representing a new class of DXR inhibitors, were synthesized, among which 5-phenylpyridin-2-ylmethylphosphonic acid possesses the most activity against E. coli DXR (EcDXR) with a K(i) of 420 nM. Structure-activity relationships (SAR) are discussed, which can be rationalized using our EcDXR:inhibitor structures, and a predictive quantitative SAR (QSAR) model is also developed. Since inhibition studies of DXR from Mycobacterium tuberculosis (MtDXR) have not been performed well, 48 EcDXR inhibitors with a broad chemical diversity were found, however, to generally exhibit considerably reduced activity against MtDXR. The crystal structure of a MtDXR:inhibitor complex reveals the flexible loop containing the residues 198-208 has no strong interactions with the 3,4-dichlorophenyl group of the inhibitor, representing a structural basis for the reduced activity. Overall, these results provide implications in the future design and development of potent DXR inhibitors.
|
Inhibition of Mycobacterium tuberculosis recombinant DXR expressed in Escherichia coli BL21 (DE3) using DXP as substrate and MgCl2 as cofactor preincubated for 10 mins
|
Mycobacterium tuberculosis
|
140.0
nM
|
|
Journal : J. Med. Chem.
Title : Inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase by lipophilic phosphonates: SAR, QSAR, and crystallographic studies.
Year : 2011
Volume : 54
Issue : 13
First Page : 4721
Last Page : 4734
Authors : Deng L, Diao J, Chen P, Pujari V, Yao Y, Cheng G, Crick DC, Prasad BV, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) is a novel target for developing new antibacterial (including antituberculosis) and antimalaria drugs. Forty-one lipophilic phosphonates, representing a new class of DXR inhibitors, were synthesized, among which 5-phenylpyridin-2-ylmethylphosphonic acid possesses the most activity against E. coli DXR (EcDXR) with a K(i) of 420 nM. Structure-activity relationships (SAR) are discussed, which can be rationalized using our EcDXR:inhibitor structures, and a predictive quantitative SAR (QSAR) model is also developed. Since inhibition studies of DXR from Mycobacterium tuberculosis (MtDXR) have not been performed well, 48 EcDXR inhibitors with a broad chemical diversity were found, however, to generally exhibit considerably reduced activity against MtDXR. The crystal structure of a MtDXR:inhibitor complex reveals the flexible loop containing the residues 198-208 has no strong interactions with the 3,4-dichlorophenyl group of the inhibitor, representing a structural basis for the reduced activity. Overall, these results provide implications in the future design and development of potent DXR inhibitors.
|
Inhibition of Mycobacterium tuberculosis recombinant DXR expressed in Escherichia coli BL21 (DE3) using DXP as substrate and Mn2+ as cofactor preincubated for 10 mins
|
Mycobacterium tuberculosis
|
80.0
nM
|
|
Journal : J. Med. Chem.
Title : Inhibition of 1-deoxy-D-xylulose-5-phosphate reductoisomerase by lipophilic phosphonates: SAR, QSAR, and crystallographic studies.
Year : 2011
Volume : 54
Issue : 13
First Page : 4721
Last Page : 4734
Authors : Deng L, Diao J, Chen P, Pujari V, Yao Y, Cheng G, Crick DC, Prasad BV, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) is a novel target for developing new antibacterial (including antituberculosis) and antimalaria drugs. Forty-one lipophilic phosphonates, representing a new class of DXR inhibitors, were synthesized, among which 5-phenylpyridin-2-ylmethylphosphonic acid possesses the most activity against E. coli DXR (EcDXR) with a K(i) of 420 nM. Structure-activity relationships (SAR) are discussed, which can be rationalized using our EcDXR:inhibitor structures, and a predictive quantitative SAR (QSAR) model is also developed. Since inhibition studies of DXR from Mycobacterium tuberculosis (MtDXR) have not been performed well, 48 EcDXR inhibitors with a broad chemical diversity were found, however, to generally exhibit considerably reduced activity against MtDXR. The crystal structure of a MtDXR:inhibitor complex reveals the flexible loop containing the residues 198-208 has no strong interactions with the 3,4-dichlorophenyl group of the inhibitor, representing a structural basis for the reduced activity. Overall, these results provide implications in the future design and development of potent DXR inhibitors.
|
Inhibition of Mycobacterium tuberculosis DXR-catalyzed NADPH-dependent rearrangement and reduction of DXP to form MEP measured every 5 secs for 500 secs by spectrophotometric analysis
|
Mycobacterium tuberculosis
|
80.0
nM
|
|
Journal : J. Med. Chem.
Title : Design, synthesis, and X-ray crystallographic studies of α-aryl substituted fosmidomycin analogues as inhibitors of Mycobacterium tuberculosis 1-deoxy-D-xylulose 5-phosphate reductoisomerase.
Year : 2011
Volume : 54
Issue : 14
First Page : 4964
Last Page : 4976
Authors : Andaloussi M, Henriksson LM, Więckowska A, Lindh M, Björkelid C, Larsson AM, Suresh S, Iyer H, Srinivasa BR, Bergfors T, Unge T, Mowbray SL, Larhed M, Jones TA, Karlén A.
Abstract : The natural antibiotic fosmidomycin acts via inhibition of 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), an essential enzyme in the non-mevalonate pathway of isoprenoid biosynthesis. Fosmidomycin is active on Mycobacterium tuberculosis DXR (MtDXR), but it lacks antibacterial activity probably because of poor uptake. α-Aryl substituted fosmidomycin analogues have more favorable physicochemical properties and are also more active in inhibiting malaria parasite growth. We have solved crystal structures of MtDXR in complex with 3,4-dichlorophenyl substituted fosmidomycin analogues; these show important differences compared to our previously described forsmidomycin-DXR complex. Our best inhibitor has an IC(50) = 0.15 μM on MtDXR but still lacked activity in a mycobacterial growth assay (MIC > 32 μg/mL). The combined results, however, provide insights into how DXR accommodates the new inhibitors and serve as an excellent starting point for the design of other novel and more potent inhibitors, particularly against pathogens where uptake is less of a problem, such as the malaria parasite.
|
Inhibition of Mycobacterium tuberculosis DXR assessed as reduction of 1-deoxy-D-xylulose 5-phosphate into 2-C-methyl-D-erythritol-4-phosphate measured for every 5 secs upto 500 secs by spectrophotometry analysis
|
Mycobacterium tuberculosis
|
80.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Substitution of the phosphonic acid and hydroxamic acid functionalities of the DXR inhibitor FR900098: an attempt to improve the activity against Mycobacterium tuberculosis.
Year : 2011
Volume : 21
Issue : 18
First Page : 5403
Last Page : 5407
Authors : Andaloussi M, Lindh M, Björkelid C, Suresh S, Wieckowska A, Iyer H, Karlén A, Larhed M.
Abstract : Two series of FR900098/fosmidomycin analogs were synthesized and evaluated for MtDXR inhibition and Mycobacterium tuberculosis whole-cell activity. The design rationale of these compounds involved the exchange of either the phosphonic acid or the hydroxamic acid part for alternative acidic and metal-coordinating functionalities. The best inhibitors provided IC(50) values in the micromolar range, with a best value of 41 μM.
|
Inhibition of Plasmodium falciparum IspC
|
Plasmodium falciparum
|
140.0
nM
|
|
Journal : J. Med. Chem.
Title : Reverse fosmidomycin derivatives against the antimalarial drug target IspC (Dxr).
Year : 2011
Volume : 54
Issue : 19
First Page : 6796
Last Page : 6802
Authors : Behrendt CT, Kunfermann A, Illarionova V, Matheeussen A, Pein MK, Gräwert T, Kaiser J, Bacher A, Eisenreich W, Illarionov B, Fischer M, Maes L, Groll M, Kurz T.
Abstract : Reverse hydroxamate-based inhibitors of IspC, a key enzyme of the non-mevalonate pathway of isoprenoid biosynthesis and a validated antimalarial target, were synthesized and biologically evaluated. The binding mode of one derivative in complex with EcIspC and a divalent metal ion was clarified by X-ray analysis. Pilot experiments have demonstrated in vivo potential.
|
Inhibition of Escherichia coli IspC
|
Escherichia coli
|
220.0
nM
|
|
Journal : J. Med. Chem.
Title : Reverse fosmidomycin derivatives against the antimalarial drug target IspC (Dxr).
Year : 2011
Volume : 54
Issue : 19
First Page : 6796
Last Page : 6802
Authors : Behrendt CT, Kunfermann A, Illarionova V, Matheeussen A, Pein MK, Gräwert T, Kaiser J, Bacher A, Eisenreich W, Illarionov B, Fischer M, Maes L, Groll M, Kurz T.
Abstract : Reverse hydroxamate-based inhibitors of IspC, a key enzyme of the non-mevalonate pathway of isoprenoid biosynthesis and a validated antimalarial target, were synthesized and biologically evaluated. The binding mode of one derivative in complex with EcIspC and a divalent metal ion was clarified by X-ray analysis. Pilot experiments have demonstrated in vivo potential.
|
Inhibition of Mycobacterium smegmatis DXR
|
Mycobacterium smegmatis
|
80.0
nM
|
|
Journal : Eur. J. Med. Chem.
Title : Growth inhibition of Mycobacterium smegmatis by prodrugs of deoxyxylulose phosphate reducto-isomerase inhibitors, promising anti-mycobacterial agents.
Year : 2012
Volume : 51
First Page : 277
Last Page : 285
Authors : Ponaire S, Zinglé C, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : Since Mycobacterium tuberculosis sets up several multiple anti-tuberculosis drug resistance mechanisms, development of new drugs with innovative target is urgent. The methylerythritol phosphate pathway (MEP) involved in the biosynthesis of essential metabolites for the survival of mycobacteria, represents such a target. Fosmidomycin 1a and FR900098 1b, two inhibitors of DXR, do not affect the viability of M. tuberculosis cells, due to a lack of uptake. To overcome the absence of the mycobacterial cell wall crossing of these compounds, we synthesized and tested the inhibition potency of acyloxymethyl phosphonate esters as prodrugs of fosmidomycin 1a, FR900098 1b and their analogs 2a and 2b on Mycobacterium smegmatis. Only the prodrugs 4b-6b inhibit the bacterial growth and could be effective anti-mycobacterial agents.
|
Inhibition of Mycobacterium smegmatis DSM 43756 ATCC 19420 N-terminal his-tagged DXR expressed in XL1-blue Escherichia coli using NADPH and DXP as substrate preincubated for 2 mins with substrate before compound addition
|
Mycobacterium smegmatis
|
510.0
nM
|
|
Journal : Eur. J. Med. Chem.
Title : Growth inhibition of Mycobacterium smegmatis by prodrugs of deoxyxylulose phosphate reducto-isomerase inhibitors, promising anti-mycobacterial agents.
Year : 2012
Volume : 51
First Page : 277
Last Page : 285
Authors : Ponaire S, Zinglé C, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : Since Mycobacterium tuberculosis sets up several multiple anti-tuberculosis drug resistance mechanisms, development of new drugs with innovative target is urgent. The methylerythritol phosphate pathway (MEP) involved in the biosynthesis of essential metabolites for the survival of mycobacteria, represents such a target. Fosmidomycin 1a and FR900098 1b, two inhibitors of DXR, do not affect the viability of M. tuberculosis cells, due to a lack of uptake. To overcome the absence of the mycobacterial cell wall crossing of these compounds, we synthesized and tested the inhibition potency of acyloxymethyl phosphonate esters as prodrugs of fosmidomycin 1a, FR900098 1b and their analogs 2a and 2b on Mycobacterium smegmatis. Only the prodrugs 4b-6b inhibit the bacterial growth and could be effective anti-mycobacterial agents.
|
Inhibition of Escherichia coli DXR using DXP as substrate preincubated for 5 mins
|
Escherichia coli
|
27.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Thermodynamic Investigation of Inhibitor Binding to 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase.
Year : 2012
Volume : 3
Issue : 6
First Page : 496
Last Page : 500
Authors : Cai G, Deng L, Fryszczyn BG, Brown NG, Liu Z, Jiang H, Palzkill T, Song Y.
Abstract : Isothermal titration calorimetry (ITC) was used to investigate the binding of six inhibitors to 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), a target for developing novel anti-infectives. The binding of hydroxamate inhibitors to E. coli DXR is Mg(2+)-dependent, highly endothermic (ΔH: 22.7-24.3 kJ/mol) and entropy-driven, while that of non-hydroxamate compounds is metal ion independent and exothermic (ΔH: -19.4- -13.8 kJ/mol), showing hydration/dehydration of the enzyme metal ion binding pocket account for the drastic ΔH change. However, for DXRs from Plasmodium falciparum and Mycobacterium tuberculosis, the binding of all inhibitors is exothermic (ΔH: -24.9 - -9.2 kJ/mol), suggesting the metal ion binding sites of these two enzymes are considerably less hydrated. The dissociation constants measured by ITC are well correlated with those obtained by enzyme inhibition assays (R(2) = 0.75). Given the rapid rise of antibiotic resistance, this work is of interest since it provides novel structural implications for rational development of potent DXR inhibitors.
|
Inhibition of His6-tagged Plasmodium falciparum DXR expressed in Escherichia coli M15 using DXP as substrate preincubated for 5 mins
|
Plasmodium falciparum
|
21.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Thermodynamic Investigation of Inhibitor Binding to 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase.
Year : 2012
Volume : 3
Issue : 6
First Page : 496
Last Page : 500
Authors : Cai G, Deng L, Fryszczyn BG, Brown NG, Liu Z, Jiang H, Palzkill T, Song Y.
Abstract : Isothermal titration calorimetry (ITC) was used to investigate the binding of six inhibitors to 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), a target for developing novel anti-infectives. The binding of hydroxamate inhibitors to E. coli DXR is Mg(2+)-dependent, highly endothermic (ΔH: 22.7-24.3 kJ/mol) and entropy-driven, while that of non-hydroxamate compounds is metal ion independent and exothermic (ΔH: -19.4- -13.8 kJ/mol), showing hydration/dehydration of the enzyme metal ion binding pocket account for the drastic ΔH change. However, for DXRs from Plasmodium falciparum and Mycobacterium tuberculosis, the binding of all inhibitors is exothermic (ΔH: -24.9 - -9.2 kJ/mol), suggesting the metal ion binding sites of these two enzymes are considerably less hydrated. The dissociation constants measured by ITC are well correlated with those obtained by enzyme inhibition assays (R(2) = 0.75). Given the rapid rise of antibiotic resistance, this work is of interest since it provides novel structural implications for rational development of potent DXR inhibitors.
|
Inhibition of Mycobacterium tuberculosis DXR using DXP as substrate preincubated for 5 mins
|
Mycobacterium tuberculosis
|
140.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Thermodynamic Investigation of Inhibitor Binding to 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase.
Year : 2012
Volume : 3
Issue : 6
First Page : 496
Last Page : 500
Authors : Cai G, Deng L, Fryszczyn BG, Brown NG, Liu Z, Jiang H, Palzkill T, Song Y.
Abstract : Isothermal titration calorimetry (ITC) was used to investigate the binding of six inhibitors to 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), a target for developing novel anti-infectives. The binding of hydroxamate inhibitors to E. coli DXR is Mg(2+)-dependent, highly endothermic (ΔH: 22.7-24.3 kJ/mol) and entropy-driven, while that of non-hydroxamate compounds is metal ion independent and exothermic (ΔH: -19.4- -13.8 kJ/mol), showing hydration/dehydration of the enzyme metal ion binding pocket account for the drastic ΔH change. However, for DXRs from Plasmodium falciparum and Mycobacterium tuberculosis, the binding of all inhibitors is exothermic (ΔH: -24.9 - -9.2 kJ/mol), suggesting the metal ion binding sites of these two enzymes are considerably less hydrated. The dissociation constants measured by ITC are well correlated with those obtained by enzyme inhibition assays (R(2) = 0.75). Given the rapid rise of antibiotic resistance, this work is of interest since it provides novel structural implications for rational development of potent DXR inhibitors.
|
Inhibition of Escherichia coli recombinant His6-tagged DXR expressed in Escherichia coli XL-1 blue using DXP as substrate after 5 mins by spectrophotometric analysis
|
Escherichia coli
|
250.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Modifications around the hydroxamic acid chelating group of fosmidomycin, an inhibitor of the metalloenzyme 1-deoxyxylulose 5-phosphate reductoisomerase (DXR).
Year : 2012
Volume : 22
Issue : 21
First Page : 6563
Last Page : 6567
Authors : Zinglé C, Kuntz L, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : Fosmidomycin derivatives in which the hydroxamic acid group has been replaced by several bidentate chelators as potential hydroxamic alternatives were prepared and tested against the DXR from Escherichia coli. These results illustrate the predominant role of the hydroxamate functional group as the most effective metal binding group in DXR inhibitors.
|
Inhibition of Escherichia coli recombinant His6-tagged DXR expressed in Escherichia coli XL-1 blue using DXP as substrate incubated for 2 mins prior to substrate addition measured after 5 mins by spectrophotometric analysis
|
Escherichia coli
|
32.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Modifications around the hydroxamic acid chelating group of fosmidomycin, an inhibitor of the metalloenzyme 1-deoxyxylulose 5-phosphate reductoisomerase (DXR).
Year : 2012
Volume : 22
Issue : 21
First Page : 6563
Last Page : 6567
Authors : Zinglé C, Kuntz L, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : Fosmidomycin derivatives in which the hydroxamic acid group has been replaced by several bidentate chelators as potential hydroxamic alternatives were prepared and tested against the DXR from Escherichia coli. These results illustrate the predominant role of the hydroxamate functional group as the most effective metal binding group in DXR inhibitors.
|
Inhibition of Arabidopsis thaliana IspC expressed in Escherichia coli after 40 min by spectrophotometric analysis
|
Arabidopsis thaliana
|
100.0
nM
|
|
Journal : Pest Manag Sci
Title : In search of new herbicidal inhibitors of the non-mevalonate pathway.
Year : 2013
Volume : 69
Issue : 5
First Page : 559
Last Page : 563
Authors : Witschel M, Röhl F, Niggeweg R, Newton T.
Abstract : The first five steps of the non-mevalonate pathway have been tested in high-throughput screening (HTS) campaigns, using enzymes of plant origin. Hit rates were in general relatively low, which could be attributed to the high polarity and charged nature of substrates and active sites of these enzymes. Still, for all the enzymes, apart from IspF (2-methylerythritol 2,4-cyclodiphosphate synthase), inhibitors could be identified with activities below 100 μM, and these were followed up to identify structure-activity relationships (SARs). For the enzyme IspD (2C-methyl-D-erythritol 4-phosphate cytidyltransferase), inhibitors with IC50 down to 35 nM were identified that also showed herbicidal activity.
|
Inhibition of Plasmodium falciparum recombinant DXR using DXP as substrate preincubated for 5 mins
|
Plasmodium falciparum
|
21.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Antimalarial and Structural Studies of Pyridine-containing Inhibitors of 1-Deoxyxylulose-5-phosphate Reductoisomerase.
Year : 2013
Volume : 4
Issue : 2
First Page : 278
Last Page : 282
Authors : Xue J, Diao J, Cai G, Deng L, Zheng B, Yao Y, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) in the non-mevalonate isoprene biosynthesis pathway is a target for developing antimalarial drugs. Fosmidomycin, a potent DXR inhibitor, showed safety as well as efficacy against P. falciparum malaria in clinical trials. Based on our previous quantitative structure activity relationship (QSAR) and crystallographic studies, several novel pyridine-containing fosmidomycin derivatives were designed, synthesized and found to be highly potent inhibitors of P. falciparum DXR (PfDXR) having Ki values of 1.9 - 13 nM, with the best one being ~11× more active than fosmidomycin. These compounds also potently block the proliferation of multi-drug resistant P. falciparum with EC50 values as low as 170 nM. A 2.3 Å crystal structure of PfDXR in complex with one of the inhibitors is reported, showing the flexible loop of the protein undergoes conformational changes upon ligand binding and a hydrogen bond and favorable hydrophobic interactions between the pyridine group and PfDXR account for the enhanced activity.
|
Inhibition of Escherichia coli recombinant DXR using DXP as substrate preincubated for 5 mins
|
Escherichia coli
|
34.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Antimalarial and Structural Studies of Pyridine-containing Inhibitors of 1-Deoxyxylulose-5-phosphate Reductoisomerase.
Year : 2013
Volume : 4
Issue : 2
First Page : 278
Last Page : 282
Authors : Xue J, Diao J, Cai G, Deng L, Zheng B, Yao Y, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) in the non-mevalonate isoprene biosynthesis pathway is a target for developing antimalarial drugs. Fosmidomycin, a potent DXR inhibitor, showed safety as well as efficacy against P. falciparum malaria in clinical trials. Based on our previous quantitative structure activity relationship (QSAR) and crystallographic studies, several novel pyridine-containing fosmidomycin derivatives were designed, synthesized and found to be highly potent inhibitors of P. falciparum DXR (PfDXR) having Ki values of 1.9 - 13 nM, with the best one being ~11× more active than fosmidomycin. These compounds also potently block the proliferation of multi-drug resistant P. falciparum with EC50 values as low as 170 nM. A 2.3 Å crystal structure of PfDXR in complex with one of the inhibitors is reported, showing the flexible loop of the protein undergoes conformational changes upon ligand binding and a hydrogen bond and favorable hydrophobic interactions between the pyridine group and PfDXR account for the enhanced activity.
|
Antimalarial activity against Plasmodium falciparum Dd2 after 3 days by SYBR green I assay
|
Plasmodium falciparum
|
440.0
nM
|
|
Journal : ACS Med. Chem. Lett.
Title : Antimalarial and Structural Studies of Pyridine-containing Inhibitors of 1-Deoxyxylulose-5-phosphate Reductoisomerase.
Year : 2013
Volume : 4
Issue : 2
First Page : 278
Last Page : 282
Authors : Xue J, Diao J, Cai G, Deng L, Zheng B, Yao Y, Song Y.
Abstract : 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) in the non-mevalonate isoprene biosynthesis pathway is a target for developing antimalarial drugs. Fosmidomycin, a potent DXR inhibitor, showed safety as well as efficacy against P. falciparum malaria in clinical trials. Based on our previous quantitative structure activity relationship (QSAR) and crystallographic studies, several novel pyridine-containing fosmidomycin derivatives were designed, synthesized and found to be highly potent inhibitors of P. falciparum DXR (PfDXR) having Ki values of 1.9 - 13 nM, with the best one being ~11× more active than fosmidomycin. These compounds also potently block the proliferation of multi-drug resistant P. falciparum with EC50 values as low as 170 nM. A 2.3 Å crystal structure of PfDXR in complex with one of the inhibitors is reported, showing the flexible loop of the protein undergoes conformational changes upon ligand binding and a hydrogen bond and favorable hydrophobic interactions between the pyridine group and PfDXR account for the enhanced activity.
|
Inhibition of N-terminal His6-tagged Escherichia coli DXR after 2 mins in presence of NADPH
|
Escherichia coli
|
32.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Synthesis of tetrazole analogues of phosphonohydroxamic acids: an attempt to improve the inhibitory activity against the DXR.
Year : 2013
Volume : 23
Issue : 6
First Page : 1643
Last Page : 1647
Authors : Nguyen-Trung AT, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : This work is focused on the design of new antimicrobial drugs and on the development of lipophilic inhibitors of the DXR, the second enzyme of the MEP pathway for the biosynthesis of isoprene units in most bacteria, by replacing the phosphonate group of fosmidomycin derivatives by a tetrazoyl moiety capable of multiple hydrogen bonding. The N- and C-substituted tetrazole analogues of phosphonohydroxamate inhibitors were synthesized and tested on the DXR of Escherichia coli. This work points out the hypothesis that the phosphonate/phosphate recognition site might be too rigid to accommodate other functional groups.
|
Inhibition of recombinant Toxoplasma gondii DXR catalytic domain (68 to 513) expressed in Escherichia coli BL21 using DXP as substrate after 10 mins by Uv-spectrophotometric analysis
|
Toxoplasma gondii
|
90.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Expression, characterization and inhibition of Toxoplasma gondii 1-deoxy-D-xylulose-5-phosphate reductoisomerase.
Year : 2013
Volume : 23
Issue : 7
First Page : 2158
Last Page : 2161
Authors : Cai G, Deng L, Xue J, Moreno SN, Striepen B, Song Y.
Abstract : The apicomplexan parasite Toxoplasma gondii, the causative agent of toxoplasmosis, is an important human pathogen. 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) in the non-mevalonate isoprene biosynthesis pathway is essential to the organism and therefore a target for developing anti-toxoplasmosis drugs. In order to find potent inhibitors, we expressed and purified recombinant T. gondii DXR (TgDXR). Biochemical properties of this enzyme were characterized and an enzyme activity/inhibition assay was developed. A collection of 11 compounds with a broad structural diversity were tested against TgDXR and several potent inhibitors were identified with Ki values as low as 48 nM. Analysis of the results as well as those of Escherichia coli and Plasmodium falciparum DXR enzymes revealed a different structure-activity relationship profile for the inhibition of TgDXR.
|
Inhibition of Mycobacterium tuberculosis DXR using DXP as substrate assessed as formation of MEP measured every 5 secs for 180 secs by spectrophotometric analysis
|
Mycobacterium tuberculosis
|
80.0
nM
|
|
Journal : J. Med. Chem.
Title : DXR inhibition by potent mono- and disubstituted fosmidomycin analogues.
Year : 2013
Volume : 56
Issue : 15
First Page : 6190
Last Page : 6199
Authors : Jansson AM, Więckowska A, Björkelid C, Yahiaoui S, Sooriyaarachchi S, Lindh M, Bergfors T, Dharavath S, Desroses M, Suresh S, Andaloussi M, Nikhil R, Sreevalli S, Srinivasa BR, Larhed M, Jones TA, Karlén A, Mowbray SL.
Abstract : The antimalarial compound fosmidomycin targets DXR, the enzyme that catalyzes the first committed step in the MEP pathway, producing the essential isoprenoid precursors, isopentenyl diphosphate and dimethylallyl diphosphate. The MEP pathway is used by a number of pathogens, including Mycobacterium tuberculosis and apicomplexan parasites, and differs from the classical mevalonate pathway that is essential in humans. Using a structure-based approach, we designed a number of analogues of fosmidomycin, including a series that are substituted in both the Cα and the hydroxamate positions. The latter proved to be a stable framework for the design of inhibitors that extend from the polar and cramped (and so not easily druggable) substrate-binding site and can, for the first time, bridge the substrate and cofactor binding sites. A number of these compounds are more potent than fosmidomycin in terms of killing Plasmodium falciparum in an in vitro assay; the best has an IC50 of 40 nM.
|
Binding affinity to Mycobacterium tuberculosis DXR assessed as dissociation constant by spectrofluorimetric analysis in presence of NADPH and MnCl2
|
Mycobacterium tuberculosis
|
40.0
nM
|
|
Journal : J. Med. Chem.
Title : DXR inhibition by potent mono- and disubstituted fosmidomycin analogues.
Year : 2013
Volume : 56
Issue : 15
First Page : 6190
Last Page : 6199
Authors : Jansson AM, Więckowska A, Björkelid C, Yahiaoui S, Sooriyaarachchi S, Lindh M, Bergfors T, Dharavath S, Desroses M, Suresh S, Andaloussi M, Nikhil R, Sreevalli S, Srinivasa BR, Larhed M, Jones TA, Karlén A, Mowbray SL.
Abstract : The antimalarial compound fosmidomycin targets DXR, the enzyme that catalyzes the first committed step in the MEP pathway, producing the essential isoprenoid precursors, isopentenyl diphosphate and dimethylallyl diphosphate. The MEP pathway is used by a number of pathogens, including Mycobacterium tuberculosis and apicomplexan parasites, and differs from the classical mevalonate pathway that is essential in humans. Using a structure-based approach, we designed a number of analogues of fosmidomycin, including a series that are substituted in both the Cα and the hydroxamate positions. The latter proved to be a stable framework for the design of inhibitors that extend from the polar and cramped (and so not easily druggable) substrate-binding site and can, for the first time, bridge the substrate and cofactor binding sites. A number of these compounds are more potent than fosmidomycin in terms of killing Plasmodium falciparum in an in vitro assay; the best has an IC50 of 40 nM.
|
Inhibition of Plasmodium falciparum DXR using DXP as substrate measured every 5 secs for 250 secs in presence of NADPH
|
Plasmodium falciparum
|
32.0
nM
|
|
Journal : J. Med. Chem.
Title : DXR inhibition by potent mono- and disubstituted fosmidomycin analogues.
Year : 2013
Volume : 56
Issue : 15
First Page : 6190
Last Page : 6199
Authors : Jansson AM, Więckowska A, Björkelid C, Yahiaoui S, Sooriyaarachchi S, Lindh M, Bergfors T, Dharavath S, Desroses M, Suresh S, Andaloussi M, Nikhil R, Sreevalli S, Srinivasa BR, Larhed M, Jones TA, Karlén A, Mowbray SL.
Abstract : The antimalarial compound fosmidomycin targets DXR, the enzyme that catalyzes the first committed step in the MEP pathway, producing the essential isoprenoid precursors, isopentenyl diphosphate and dimethylallyl diphosphate. The MEP pathway is used by a number of pathogens, including Mycobacterium tuberculosis and apicomplexan parasites, and differs from the classical mevalonate pathway that is essential in humans. Using a structure-based approach, we designed a number of analogues of fosmidomycin, including a series that are substituted in both the Cα and the hydroxamate positions. The latter proved to be a stable framework for the design of inhibitors that extend from the polar and cramped (and so not easily druggable) substrate-binding site and can, for the first time, bridge the substrate and cofactor binding sites. A number of these compounds are more potent than fosmidomycin in terms of killing Plasmodium falciparum in an in vitro assay; the best has an IC50 of 40 nM.
|
Inhibition of Escherichia coli DXR using DXP as substrate measured every 5 secs for 250 secs in presence of NADPH
|
Escherichia coli
|
50.0
nM
|
|
Journal : J. Med. Chem.
Title : DXR inhibition by potent mono- and disubstituted fosmidomycin analogues.
Year : 2013
Volume : 56
Issue : 15
First Page : 6190
Last Page : 6199
Authors : Jansson AM, Więckowska A, Björkelid C, Yahiaoui S, Sooriyaarachchi S, Lindh M, Bergfors T, Dharavath S, Desroses M, Suresh S, Andaloussi M, Nikhil R, Sreevalli S, Srinivasa BR, Larhed M, Jones TA, Karlén A, Mowbray SL.
Abstract : The antimalarial compound fosmidomycin targets DXR, the enzyme that catalyzes the first committed step in the MEP pathway, producing the essential isoprenoid precursors, isopentenyl diphosphate and dimethylallyl diphosphate. The MEP pathway is used by a number of pathogens, including Mycobacterium tuberculosis and apicomplexan parasites, and differs from the classical mevalonate pathway that is essential in humans. Using a structure-based approach, we designed a number of analogues of fosmidomycin, including a series that are substituted in both the Cα and the hydroxamate positions. The latter proved to be a stable framework for the design of inhibitors that extend from the polar and cramped (and so not easily druggable) substrate-binding site and can, for the first time, bridge the substrate and cofactor binding sites. A number of these compounds are more potent than fosmidomycin in terms of killing Plasmodium falciparum in an in vitro assay; the best has an IC50 of 40 nM.
|
Inhibition of Mycobacterium tuberculosis IspC by photometric assay
|
Mycobacterium tuberculosis
|
230.0
nM
|
|
Journal : J. Med. Chem.
Title : IspC as target for antiinfective drug discovery: synthesis, enantiomeric separation, and structural biology of fosmidomycin thia isosters.
Year : 2013
Volume : 56
Issue : 20
First Page : 8151
Last Page : 8162
Authors : Kunfermann A, Lienau C, Illarionov B, Held J, Gräwert T, Behrendt CT, Werner P, Hähn S, Eisenreich W, Riederer U, Mordmüller B, Bacher A, Fischer M, Groll M, Kurz T.
Abstract : The emergence and spread of multidrug-resistant pathogens are widely believed to endanger human health. New drug targets and lead compounds exempt from cross-resistance with existing drugs are urgently needed. We report on the synthesis and properties of "reverse" thia analogs of fosmidomycin, which inhibit the first committed enzyme of a metabolic pathway that is essential for the causative agents of tuberculosis and malaria but is absent in the human host. Notably, IspC displays a high level of enantioselectivity for an α-substituted fosmidomycin derivative.
|
Inhibition of Escherichia coli IspC by photometric assay
|
Escherichia coli
|
221.0
nM
|
|
Journal : J. Med. Chem.
Title : IspC as target for antiinfective drug discovery: synthesis, enantiomeric separation, and structural biology of fosmidomycin thia isosters.
Year : 2013
Volume : 56
Issue : 20
First Page : 8151
Last Page : 8162
Authors : Kunfermann A, Lienau C, Illarionov B, Held J, Gräwert T, Behrendt CT, Werner P, Hähn S, Eisenreich W, Riederer U, Mordmüller B, Bacher A, Fischer M, Groll M, Kurz T.
Abstract : The emergence and spread of multidrug-resistant pathogens are widely believed to endanger human health. New drug targets and lead compounds exempt from cross-resistance with existing drugs are urgently needed. We report on the synthesis and properties of "reverse" thia analogs of fosmidomycin, which inhibit the first committed enzyme of a metabolic pathway that is essential for the causative agents of tuberculosis and malaria but is absent in the human host. Notably, IspC displays a high level of enantioselectivity for an α-substituted fosmidomycin derivative.
|
Inhibition of Mycobacterium tuberculosis Dxr
|
Mycobacterium tuberculosis
|
440.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : The effect of chain length and unsaturation on Mtb Dxr inhibition and antitubercular killing activity of FR900098 analogs.
Year : 2014
Volume : 24
Issue : 2
First Page : 649
Last Page : 653
Authors : Jackson ER, San Jose G, Brothers RC, Edelstein EK, Sheldon Z, Haymond A, Johny C, Boshoff HI, Couch RD, Dowd CS.
Abstract : Inhibition of the nonmevalonate pathway (NMP) of isoprene biosynthesis has been examined as a source of new antibiotics with novel mechanisms of action. Dxr is the best studied of the NMP enzymes and several reports have described potent Dxr inhibitors. Many of these compounds are structurally related to natural products fosmidomycin and FR900098, each bearing retrohydroxamate and phosphonate groups. We synthesized a series of compounds with two to five methylene units separating these groups to examine what linker length was optimal and tested for inhibition against Mtb Dxr. We synthesized ethyl and pivaloyl esters of these compounds to increase lipophilicity and improve inhibition of Mtb growth. Our results show that propyl or propenyl linker chains are optimal. Propenyl analog 22 has an IC50 of 1.07 μM against Mtb Dxr. The pivaloyl ester of 22, compound 26, has an MIC of 9.4 μg/mL, representing a significant improvement in antitubercular potency in this class of compounds.
|
Inhibition of His-tagged Mycobacterium tuberculosis DXR expressed in Escherichia coli BL21(DE3) using DXP as substrate assessed as oxidation of NADPH preincubated for 10 mins followed by NADPH and substrate addition by spectrophotometric analysis
|
Mycobacterium tuberculosis
|
310.0
nM
|
|
Journal : MedChemComm
Title : Design of Potential Bisubstrate Inhibitors against Mycobacterium tuberculosis (Mtb) 1-Deoxy-D-Xylulose 5-Phosphate Reductoisomerase (Dxr)-Evidence of a Novel Binding Mode.
Year : 2013
Volume : 4
Issue : 7
First Page : 1099
Last Page : 1104
Authors : San Jose G, Jackson ER, Uh E, Johny C, Haymond A, Lundberg L, Pinkham C, Kehn-Hall K, Boshoff HI, Couch RD, Dowd CS.
Abstract : In most bacteria, the nonmevalonate pathway is used to synthesize isoprene units. Dxr, the second step in the pathway, catalyzes the NADPH-dependent reductive isomerization of 1-deoxy-D-xylulose-5-phosphate (DXP) to 2-C-methyl-D-erythritol-4-phosphate (MEP). Dxr is inhibited by natural products fosmidomycin and FR900098, which bind in the DXP binding site. These compounds, while potent inhibitors of Dxr, lack whole cell activity against Mycobacterium tuberculosis (Mtb) due to their polarity. Our goal was to use the Mtb Dxr-fosmidomycin co-crystal structure to design bisubstrate ligands to bind to both the DXP and NADPH sites. Such compounds would be expected to demonstrate improved whole cell activity due to increased lipophilicity. Two series of compounds were designed and synthesized. Compounds from both series inhibited Mtb Dxr. The most potent compound (8) has an IC50 of 17.8 µM. Analysis shows 8 binds to Mtb Dxr via a novel, non-bisubstrate mechanism. Further, the diethyl ester of 8 inhibits Mtb growth making this class of compounds interesting lead molecules in the search for new antitubercular agents.
|
Inhibition of Escherichia coli IspC
|
Escherichia coli
|
35.0
nM
|
|
Journal : MedChemComm
Title : The MEP pathway and the development of inhibitors as potential anti-infective agents
Year : 2012
Volume : 3
Issue : 4
First Page : 418
Last Page : 433
Authors : Hale I, O'Neill PM, Berry NG, Odom A, Sharma R
|
Inhibition of Escherichia coli Dxr
|
Escherichia coli
|
32.0
nM
|
|
Journal : J. Med. Chem.
Title : Development of inhibitors of the 2C-methyl-D-erythritol 4-phosphate (MEP) pathway enzymes as potential anti-infective agents.
Year : 2014
Volume : 57
Issue : 23
First Page : 9740
Last Page : 9763
Authors : Masini T, Hirsch AK.
Abstract : Important pathogens such as Mycobacterium tuberculosis and Plasmodium falciparum, the causative agents of tuberculosis and malaria, respectively, and plants, utilize the 2C-methyl-D-erythritol 4-phosphate (MEP, 5) pathway for the biosynthesis of isopentenyl diphosphate (1) and dimethylallyl diphosphate (2), the universal precursors of isoprenoids, while humans exclusively utilize the alternative mevalonate pathway for the synthesis of 1 and 2. This distinct distribution, together with the fact that the MEP pathway is essential in numerous organisms, makes the enzymes of the MEP pathway attractive drug targets for the development of anti-infective agents and herbicides. Herein, we review the inhibitors reported over the past 2 years, in the context of the most important older developments and with a particular focus on the results obtained against enzymes of pathogenic organisms. We will also discuss new discoveries in terms of structural and mechanistic features, which can help to guide a rational development of inhibitors.
|
Inhibition of His-tagged Escherichia coli DXR pre-incubated for 2 mins before reaction initiation in presence of 160 uM NADPH in absence of 0.01% Triton X100
|
Escherichia coli
|
32.0
nM
|
|
Journal : Bioorg. Med. Chem.
Title : Catechol-rhodanine derivatives: Specific and promiscuous inhibitors of Escherichia coli deoxyxylulose phosphate reductoisomerase (DXR).
Year : 2014
Volume : 22
Issue : 14
First Page : 3713
Last Page : 3719
Authors : Zinglé C, Tritsch D, Grosdemange-Billiard C, Rohmer M.
Abstract : To develop more effective inhibitors than fosmidomycin, a natural compound which inhibits the deoxyxylulose 5-phosphate reductoisomerase (DXR), the second enzyme of the MEP pathway, we designed molecules possessing on the one hand a catechol that is able to chelate the magnesium dication and on the other hand a group able to occupy the NADPH recognition site. Catechol-rhodanine derivatives (1-6) were synthesized and their potential inhibition was tested on the DXR of Escherichia coli. For the inhibitors 1 and 2, the presence of detergent in the enzymatic assays led to a dramatic decrease of the inhibition suggesting, that these compounds are rather promiscuous inhibitors. The compounds 4 and 5 kept their inhibition capacity in the presence of Triton X100 and could be considered as specific inhibitors of DXR. Compound 4 showed antimicrobial activity against Escherichia coli. The only partial protection of NADPH against the inhibition suggested that the catechol-rhodanine derivatives did not settle in the coenzyme binding site. This paper points out the necessity to include a detergent in the DXR enzymatic assays to avoid false positive when putative hydrophobic inhibitors are tested and especially when the IC50, are in the micromolar range.
|
Antimalarial activity against MRA-150, chloroquine, pyrimethamine, mefloquine-resistant asexual blood stage Plasmodium falciparum Dd2 after 72 hrs by SYBR green assay
|
Plasmodium falciparum Dd2
|
880.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Determination of the active stereoisomer of the MEP pathway-targeting antimalarial agent MMV008138, and initial structure-activity studies.
Year : 2015
Volume : 25
Issue : 7
First Page : 1515
Last Page : 1519
Authors : Yao ZK, Krai PM, Merino EF, Simpson ME, Slebodnick C, Cassera MB, Carlier PR.
Abstract : Compounds that target isoprenoid biosynthesis in Plasmodium falciparum could be a welcome addition to malaria chemotherapy, since the methylerythritol phosphate (MEP) pathway used by the parasite is not present in humans. We previously reported that MMV008138 targets the apicoplast of P. falciparum and that its target in the MEP pathway differs from that of Fosmidomycin. In this Letter, we determine that the active stereoisomer of MMV008138 is 4a, which is (1R,3S)-configured. 2',4'-Disubstitution of the D ring was also found to be crucial for inhibition of the parasite growth. Limited variation of the C3-carboxylic acid substituent was carried out, and methylamide derivative 8a was found to be more potent than 4a; other amides, acylhydrazines, and esters were less potent. Finally, lead compounds 4a, 4e, 4f, 4h, 8a, and 8e did not inhibit growth of Escherichia coli, suggesting that protozoan-selective inhibition of the MEP pathway of P. falciparum can be achieved.
|
Antimalarial activity against chloroquine and mefloquine-resistant Plasmodium falciparum Dd2 after 48 hrs by SYBR Green I assay
|
Plasmodium falciparum Dd2
|
810.0
nM
|
|
Journal : Bioorg. Med. Chem. Lett.
Title : Synthesis and antimalarial evaluation of prodrugs of novel fosmidomycin analogues.
Year : 2015
Volume : 25
Issue : 10
First Page : 2112
Last Page : 2116
Authors : Faísca Phillips AM, Nogueira F, Murtinheira F, Barros MT.
Abstract : The continuous development of drug resistance by Plasmodium falciparum, the agent responsible for the most severe forms of malaria, creates the need for the development of novel drugs to fight this disease. Fosmidomycin is an effective antimalarial and potent antibiotic, known to act by inhibiting the enzyme 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR), essential for the synthesis of isoprenoids in eubacteria and plasmodia, but not in humans. In this study, novel constrained cyclic prodrug analogues of fosmidomycin were synthesized. One, in which the hydroxamate function is incorporated into a six-membered ring, was found have higher antimalarial activity than fosmidomycin against the chloroquine and mefloquine resistant P. falciparum Dd2 strain. In addition, it showed very low cytotoxicity against cultured human cells.
|
Inhibition of recombinant Plasmodium falciparum IspC using [3,4,5-13C3]1-Deoxy-D-xylulose 5-phosphate substrate by photometric assay
|
Plasmodium falciparum
|
160.0
nM
|
|
Journal : J. Med. Chem.
Title : Prodrugs of reverse fosmidomycin analogues.
Year : 2015
Volume : 58
Issue : 4
First Page : 2025
Last Page : 2035
Authors : Brücher K, Gräwert T, Konzuch S, Held J, Lienau C, Behrendt C, Illarionov B, Maes L, Bacher A, Wittlin S, Mordmüller B, Fischer M, Kurz T.
Abstract : Fosmidomycin inhibits IspC (Dxr, 1-deoxy-d-xylulose 5-phosphate reductoisomerase), a key enzyme in nonmevalonate isoprenoid biosynthesis that is essential in Plasmodium falciparum. The drug has been used successfully to treat malaria patients in clinical studies, thus validating IspC as an antimalarial target. However, improvement of the drug's pharmacodynamics and pharmacokinetics is desirable. Here, we show that the conversion of the phosphonate moiety into acyloxymethyl and alkoxycarbonyloxymethyl groups can increase the in vitro activity against asexual blood stages of P. falciparum by more than 1 order of magnitude. We also synthesized double prodrugs by additional esterification of the hydroxamate moiety. Prodrugs with modified hydroxamate moieties are subject to bioactivation in vitro. All prodrugs demonstrated improved antiplasmodial in vitro activity. Selected prodrugs and parent compounds were also tested for their cytotoxicity toward HeLa cells and in vivo in a Plasmodium berghei malaria model as well as in the SCID mouse P. falciparum model.
|
Antiplasmodial activity against blood stage forms of multidrug-resistant Plasmodium falciparum Dd2 incubated for 3 days by HRP2 detection based ELISA method
|
Plasmodium falciparum Dd2
|
810.0
nM
|
|
Journal : J. Med. Chem.
Title : Prodrugs of reverse fosmidomycin analogues.
Year : 2015
Volume : 58
Issue : 4
First Page : 2025
Last Page : 2035
Authors : Brücher K, Gräwert T, Konzuch S, Held J, Lienau C, Behrendt C, Illarionov B, Maes L, Bacher A, Wittlin S, Mordmüller B, Fischer M, Kurz T.
Abstract : Fosmidomycin inhibits IspC (Dxr, 1-deoxy-d-xylulose 5-phosphate reductoisomerase), a key enzyme in nonmevalonate isoprenoid biosynthesis that is essential in Plasmodium falciparum. The drug has been used successfully to treat malaria patients in clinical studies, thus validating IspC as an antimalarial target. However, improvement of the drug's pharmacodynamics and pharmacokinetics is desirable. Here, we show that the conversion of the phosphonate moiety into acyloxymethyl and alkoxycarbonyloxymethyl groups can increase the in vitro activity against asexual blood stages of P. falciparum by more than 1 order of magnitude. We also synthesized double prodrugs by additional esterification of the hydroxamate moiety. Prodrugs with modified hydroxamate moieties are subject to bioactivation in vitro. All prodrugs demonstrated improved antiplasmodial in vitro activity. Selected prodrugs and parent compounds were also tested for their cytotoxicity toward HeLa cells and in vivo in a Plasmodium berghei malaria model as well as in the SCID mouse P. falciparum model.
|
Antiplasmodial activity against blood stage forms of chloroquine-sensitive Plasmodium falciparum 3D7 incubated for 3 days by HRP2 detection based ELISA method
|
Plasmodium falciparum 3D7
|
880.0
nM
|
|
Journal : J. Med. Chem.
Title : Prodrugs of reverse fosmidomycin analogues.
Year : 2015
Volume : 58
Issue : 4
First Page : 2025
Last Page : 2035
Authors : Brücher K, Gräwert T, Konzuch S, Held J, Lienau C, Behrendt C, Illarionov B, Maes L, Bacher A, Wittlin S, Mordmüller B, Fischer M, Kurz T.
Abstract : Fosmidomycin inhibits IspC (Dxr, 1-deoxy-d-xylulose 5-phosphate reductoisomerase), a key enzyme in nonmevalonate isoprenoid biosynthesis that is essential in Plasmodium falciparum. The drug has been used successfully to treat malaria patients in clinical studies, thus validating IspC as an antimalarial target. However, improvement of the drug's pharmacodynamics and pharmacokinetics is desirable. Here, we show that the conversion of the phosphonate moiety into acyloxymethyl and alkoxycarbonyloxymethyl groups can increase the in vitro activity against asexual blood stages of P. falciparum by more than 1 order of magnitude. We also synthesized double prodrugs by additional esterification of the hydroxamate moiety. Prodrugs with modified hydroxamate moieties are subject to bioactivation in vitro. All prodrugs demonstrated improved antiplasmodial in vitro activity. Selected prodrugs and parent compounds were also tested for their cytotoxicity toward HeLa cells and in vivo in a Plasmodium berghei malaria model as well as in the SCID mouse P. falciparum model.
|
Inhibition of recombinant Escherichia coli Dxr expressed in Escherichia coli BL21 CodonPlus (DE3)-RIL cells using DOXP as substrate assessed as NADPH oxidation preincubated for 5 mins with NADPH followed by substrate addition by spectrophotometric analysis
|
Escherichia coli
|
438.0
nM
|
|
Journal : J. Med. Chem.
Title : Synthesis and bioactivity of β-substituted fosmidomycin analogues targeting 1-deoxy-D-xylulose-5-phosphate reductoisomerase.
Year : 2015
Volume : 58
Issue : 7
First Page : 2988
Last Page : 3001
Authors : Chofor R, Sooriyaarachchi S, Risseeuw MD, Bergfors T, Pouyez J, Johny C, Haymond A, Everaert A, Dowd CS, Maes L, Coenye T, Alex A, Couch RD, Jones TA, Wouters J, Mowbray SL, Van Calenbergh S.
Abstract : Blocking the 2-C-methyl-d-erythrithol-4-phosphate (MEP) pathway for isoprenoid biosynthesis offers interesting prospects for inhibiting Plasmodium or Mycobacterium spp. growth. Fosmidomycin (1) and its homologue FR900098 (2) potently inhibit 1-deoxy-d-xylulose-5-phosphate reductoisomerase (Dxr), a key enzyme in this pathway. Here we introduced aryl or aralkyl substituents at the β-position of the hydroxamate analogue of 2. While direct addition of a β-aryl moiety resulted in poor inhibition, longer linkers between the carbon backbone and the phenyl ring were generally associated with better binding to the enzymes. X-ray structures of the parasite Dxr-inhibitor complexes show that the "longer" compounds generate a substantially different flap structure, in which a key tryptophan residue is displaced, and the aromatic group of the ligand lies between the tryptophan and the hydroxamate's methyl group. Although the most promising new Dxr inhibitors lack activity against Escherichia coli and Mycobacterium smegmatis, they proved to be highly potent inhibitors of Plasmodium falciparum in vitro growth.
|
Inhibition of recombinant Plasmodium falciparum Dxr expressed in Escherichia coli C43(DE3) using DOXP as substrate assessed as NADPH oxidation by spectrophotometric analysis
|
Plasmodium falciparum
|
36.0
nM
|
|
Journal : J. Med. Chem.
Title : Synthesis and bioactivity of β-substituted fosmidomycin analogues targeting 1-deoxy-D-xylulose-5-phosphate reductoisomerase.
Year : 2015
Volume : 58
Issue : 7
First Page : 2988
Last Page : 3001
Authors : Chofor R, Sooriyaarachchi S, Risseeuw MD, Bergfors T, Pouyez J, Johny C, Haymond A, Everaert A, Dowd CS, Maes L, Coenye T, Alex A, Couch RD, Jones TA, Wouters J, Mowbray SL, Van Calenbergh S.
Abstract : Blocking the 2-C-methyl-d-erythrithol-4-phosphate (MEP) pathway for isoprenoid biosynthesis offers interesting prospects for inhibiting Plasmodium or Mycobacterium spp. growth. Fosmidomycin (1) and its homologue FR900098 (2) potently inhibit 1-deoxy-d-xylulose-5-phosphate reductoisomerase (Dxr), a key enzyme in this pathway. Here we introduced aryl or aralkyl substituents at the β-position of the hydroxamate analogue of 2. While direct addition of a β-aryl moiety resulted in poor inhibition, longer linkers between the carbon backbone and the phenyl ring were generally associated with better binding to the enzymes. X-ray structures of the parasite Dxr-inhibitor complexes show that the "longer" compounds generate a substantially different flap structure, in which a key tryptophan residue is displaced, and the aromatic group of the ligand lies between the tryptophan and the hydroxamate's methyl group. Although the most promising new Dxr inhibitors lack activity against Escherichia coli and Mycobacterium smegmatis, they proved to be highly potent inhibitors of Plasmodium falciparum in vitro growth.
|
Inhibition of Escherichia coli His6-tagged DXR preincubated for 2 mins in presence of NADPH followed by DXP substrate addition
|
Escherichia coli
|
42.0
nM
|
|
Journal : Bioorg Med Chem
Title : Synthesis and biological evaluation of phosphate isosters of fosmidomycin and analogs as inhibitors of Escherichia coli and Mycobacterium smegmatis 1-deoxyxylulose 5-phosphate reductoisomerases.
Year : 2017
Volume : 25
Issue : 2
First Page : 684
Last Page : 689
Authors : Munier M, Tritsch D, Krebs F, Esque J, Hemmerlin A, Rohmer M, Stote RH, Grosdemange-Billiard C.
Abstract : Hydroxamate analogs of fosfoxacin, the phosphate homolog of fosmidomycin, have been synthesized and their activity tested on Escherichia coli and Mycobacterium smegmatis DXRs. Except for compound 4b, the IC50 values of phosphate derivatives are approximately 10-fold higher than those of the corresponding phosphonates. Although their inhibitory activity on Escherichia coli DXR is less efficient than their phosphonate analogs, we report the ability of phosphate compounds to inhibit the growth of Escherichia coli. This work points out that the uptake of fosfoxacin and its analogs is taking place via the GlpT and UhpT transporters. As expected, these compounds are inefficient to inhibit the growth of M. smegmatis growth inhibition probably due to a lack of uptake.
|
Antimalarial activity against Plasmodium falciparum Dd2 assessed as reduction in parasite viability by [3H]-hypoxanthine incorporation assay
|
Plasmodium falciparum
|
290.0
nM
|
|
Journal : J Med Chem
Title : Antimalarial Chemotherapy: Natural Product Inspired Development of Preclinical and Clinical Candidates with Diverse Mechanisms of Action.
Year : 2016
Volume : 59
Issue : 12
First Page : 5587
Last Page : 5603
Authors : Fernández-Álvaro E, Hong WD, Nixon GL, O'Neill PM, Calderón F.
Abstract : Natural products have played a pivotal role in malaria chemotherapy progressing from quinine and artemisinin to ozonide-based compounds. Many of these natural products have served as template for the design and development of antimalarial drugs currently in the clinic or in the development phase. In this review, we will detail those privileged scaffolds that have guided medicinal chemistry efforts yielding molecules that have reached the clinic.
