Download Hi-Res ImageDownload to MS-PowerPointCite This:J. Am. Chem. Soc. 2018, 140, 7, 2537-2545

ArticleDecember 22, 2017

Glycomimetic, Orally Bioavailable LecB Inhibitors Block Biofilm Formation of Pseudomonas aeruginosa
Click to copy article linkArticle link copied!

View Author Information

Open PDF Supporting Information (1)

Journal of the American Chemical Society

Cite this: J. Am. Chem. Soc. 2018, 140, 7, 2537–2545

Click to copy citationCitation copied!

https://doi.org/10.1021/jacs.7b11133

Published December 22, 2017

Request reuse permissions

Abstract

Click to copy section linkSection link copied!

The opportunistic Gram-negative bacterium Pseudomonas aeruginosa is a leading pathogen for infections of immuno-compromised patients and those suffering from cystic fibrosis. Its ability to switch from planktonic life to aggregates, forming the so-called biofilms, is a front-line mechanism of antimicrobial resistance. The bacterial carbohydrate-binding protein LecB is an integral component and necessary for biofilm formation. Here, we report a new class of drug-like low molecular weight inhibitors of the lectin LecB with nanomolar affinities and excellent receptor binding kinetics and thermodynamics. This class of glycomimetic inhibitors efficiently blocked biofilm formation of P. aeruginosain vitro while the natural monovalent carbohydrate ligands failed. Furthermore, excellent selectivity and pharmacokinetic properties were achieved. Notably, two compounds showed good oral bioavailability, and high compound concentrations in plasma and urine were achieved in vivo.

Subjects

Note Added after ASAP Publication

This paper was published January 22, 2018. In the article and supporting information, compound 2a related to the X-ray structure has been corrected to 3a. The revised version was posted on February 21, 2018.

Introduction

Click to copy section linkSection link copied!

The Gram-negative bacterium Pseudomonas aeruginosa has become a severe threat for hospitalized immuno-compromised patients and is now a critical priority 1 pathogen as stated by the WHO in 2017. (1-4) In persons suffering from cystic fibrosis, chronic infections with this pathogen lead to recurrent pneumonia, sepsis, and lung damage. (5) Infections of various organs and tissues by this pathogen, such as chronic wound (6) or catheter-associated urinary tract infections (7) (caUTI), impose severe clinical challenges which further result from the bacterium’s intrinsic drug resistance and additional acquired resistances often leading to multi-drug-resistant or extremely drug-resistant strains (MDR, XDR). (8) Furthermore, the antimicrobial tolerance of P. aeruginosa is enhanced through formation of biofilms, a self-made environment that protects bacteria against host immune defense and antibiotic treatment. (9, 10) Triggered by the fact that bacteria inside a biofilm are up to 1000-fold more resistant toward antibiotics, (9) the targeting of biofilm formation as therapeutic approach emerged in order to overcome the resistance problem (reviewed in refs 11 and 12).

The two quorum-sensing-regulated (13) virulence factors LecA (14) and LecB (15) (formerly called PA-IL and PA-IIL) are crucial for biofilm formation by P. aeruginosa. These two carbohydrate-binding proteins, i.e., lectins, were initially isolated from the clinical isolate PAO1 by Gilboa-Garber et al. (16) Problematic for therapy, P. aeruginosa shows a high genomic diversity among different isolates. (17, 18) While LecA is relatively conserved among strains, the genomic diversity of LecB varies, and clinical isolates can be grouped into either PAO1- or PA14-like LecB sequence families, with both types covering approximately 50% each of the clinical isolates. (19, 20) However, we could show that despite numerous sequence variations in the two LecB variants, they possess comparable binding specificity which is a prerequisite for targeting a broad range of clinical isolates with one single drug. (20)

The exact mechanism of the lectins’ role in biofilm formation is not fully understood. It is assumed, that these tetravalent lectins act as bridging agents to enable glycan-mediated bacterial adhesion to the cellular glycocalix of the host’s tissue. In addition, they are believed to mediate contact between bacterial cells through cell-surface glycoconjugates and with the bacterially produced multivalent exopolysaccharides for the establishment of bacterial aggregates and biofilms. (11) This biofilm-reinforcing property of the lectins, which results from its multivalency, can be blocked by inhibition of the carbohydrate binding sites.

In a mouse infection model, LecB was shown to be the major determinant for bacterial lung colonization. (21, 22) Furthermore, this protein influences signaling pathways in human lung epithelial cells leading to β-catenin degradation, a host protein that is crucially involved in tissue repair. (23) On a structural level, LecB forms noncovalent homotetramers, and each monomer contains two Ca²⁺-ions, (20, 24) which mediate the binding to its carbohydrate ligands, l-fucose and d-mannose. It has been shown in humans and mice that inhalation of millimolar concentrations of these natural sugars contributed to clearance of bacteria from the lungs and, furthermore, showed synergistic activities with coadministered antibiotics. (25, 26) Due to higher affinities toward fucosides over mannosides, research focused on fucose-based inhibitors with multivalent presentation to further increase avidity. (27, 28) This approach yielded multivalent glycopeptide dendrimer structures with the ability to efficiently inhibit the formation of and disperse established biofilms of P. aeruginosa. (29) Interestingly, in another case of multivalent fucosides, despite nanomolar affinities for LecB, millimolar concentrations (5 mM) have been reported as necessary for efficient inhibition of biofilm formation, and compounds were inactive at 100 μM. (21) In the same study, these multivalent inhibitors were even shown to promote bacterial aggregation. Thus, it is conceivable that some multivalent lectin inhibitors can act as mimics of the bacterial exopolysaccharides and therefore unintentionally contribute to biofilm integrity rather than to its desired disintegration.

In order to circumvent such a detrimental biofilm-stabilizing effect of multivalent ligands through cross-linking of the tetravalent lectins inside the biofilm, we have focused on monovalent low molecular weight inhibitors of LecB. (30-34) In addition, only small molecules can be developed into drug-like molecules with favorable pharmakokinetic properties, i.e., oral application with systemic drug bioavailability, which is necessary to reach all possible types of the various organ and tissue infections elicited by P. aeruginosa. Thus, small monovalent molecules hold the premise to circumvent both, biofilm-reinforcing effects and the restriction to topical applications for the multivalent systems.

Here, we report two types of monovalent glycomimetic LecB inhibitors, cinnamides 6 and sulfonamides 7. These new low molecular weight C-glycosidic LecB inhibitors showed nanomolar potency against LecB with excellent target selectivity, toxicity, and in vitro ADME properties. Notably, the glycomimetic sulfonamides developed here were potent inhibitors of biofilm formation in stark contrast to the ineffective native carbohydrates. Finally, a murine in vivo pharmacokinetic study revealed the suitability of these glycomimetics for oral application in a proof-of-concept infection model.

Results and Discussion

Click to copy section linkSection link copied!

Design and Synthesis of C-Glycoside LecB Inhibitors

We have previously modified the rather weak LecB ligand methyl α-d-mannoside (1) into derivatives with various aromatic substituents attached to C-6, which increased potency up to a factor of 20 and addressed a new subsite at the target, e.g., 2a–c and 3a–c, Figures 1 and 2. (30, 32) In contrast to rapidly dissociating 1 (t_1/2 = 45 s), these compounds showed extended receptor residence times with complex half-lives in the 5–20 min range, revealing the molecular basis for the increased potency. This property is an important requirement for in vivo efficacy. In order to further optimize target interactions and increase stability toward hydrolases, we merged functional groups establishing attractive interactions with LecB from the natural ligands into one molecule (1 → 5, Figure 1). (31) The equatorial methyl group in 5 resulted in C-glycosides with 4- to 6-fold enhanced affinities originating from the reported lipophilic contact of the l-fucose methyl group with the lectin. (20, 24, 35) Despite these numerous improvements, the previously reported molecules display only moderate affinities for LecB in the range of 3.3–16 μM, which is insufficient for application. Now, we aimed to combine the individually optimized structures of mannose-derivatives 2a/3a and C-glycoside 5 into monovalent drug-like molecules 6 and 7 to increase affinity toward the target and assess their biological properties. Furthermore, these new glycomimetic structures, 6 and 7, are expected to be more stable against host metabolism compared to their glycosidic precursors, resulting from the absence of a labile glycosidic linkage. This design should enable higher compound concentrations in vivo.

Figure 1. Rational design of 6 and 7 as monovalent C-glycosidic glycomimetic LecB inhibitors. Derivatives of methyl α-d-mannoside 1–5 and their inhibitory potency for the binding with LecB_PAO1. (30, 31, 35) Moieties colored in blue increase potency by improving binding kinetics. (30, 32) The orange colored methyl group originating from l-fucosides enhances binding to LecB through a lipophilic interaction. (31, 35)

Figure 2. Competitive binding assay of inhibitors with LecB_PAO1 and LecB_PA14. Means and standard deviations were determined from a minimum of three independent measurements. IC₅₀ values for 1 with both LecB variants and 2a–c, 3a,c and 5 with LecB_PAO1 were previously published. (20, 30-32) One representative titration with LecB_PA14 is depicted for reference pairs 2b/6b and 3a/7a. Gray arrows indicate the increase in activity for C-glycosides.

To access β-C-glycosides 6 and 7, we utilized the previously described synthesis of amine 10, (31) where l-fucose (8) was transformed to β-C-glycoside 9 by Henry addition of nitromethane under basic conditions followed by a reduction to yield amine 10 (Scheme 1). Then, fucose/mannose hybrid-type structures 6 and 7 bearing amide or sulfonamide substituents, respectively, were generated by coupling with different electrophiles.

Scheme 1. Synthesis of Structures 6a,b and 7a,b^a

Besides the amide-bridged cinnamic acid moiety as present in 2a and 6a or the sulfonamide-bridged trimethylphenyl moiety in 3a and 7a, a thiophenesulfonamide was introduced in mannoside 3b and C-glycoside 7b as a bioisoster of the potent phenylsulfonamide 3c (30) in the mannose-series. The final coupling step yielded amides 6a,b and sulfonamides 7a,b in moderate to good yields (33–72%, over 2 steps).

Improved Binding Properties of C-Glycoside Structures toward LecB

Derivatives 6a,b and 7a,b were then analyzed for their capacity to inhibit the two representative lectin variants of the two clinically occurring strain clades, LecB_PAO1 and LecB_PA14, based on previously established competitive binding assays (20, 30) (Figure 2). Furthermore, all other glycomimetics (i.e., 2a–c, 3a–c, 5) were assayed for the first time also toward LecB_PA14 and showed stronger binding to the PA14 variant compared to that toward LecB_PAO1, with the exception of cinnamides 2a,b. In general, enhanced affinities were detected for C-glycoside derivatives 6a,b and 7a,b compared to those of their respective mannose analogs 2a,b and 3a,b. Cinnamide 6a (IC₅₀[LecB_PAO1] 4.21 μM; IC₅₀[LecB_PA14] 2.49 μM) showed 9- to 16-fold higher affinities toward both LecB types compared to that of its mannose reference 2a (IC₅₀[LecB_PAO1] 37 μM; (30) IC₅₀[LecB_PA14] 39 μM). Sulfonamide 7a (IC₅₀[LecB_PAO1] 0.97 μM; IC₅₀[LecB_PA14] 0.34 μM) was a 3- to 5-fold stronger ligand of LecB compared to that of corresponding mannoside 3a, exceeding the potency of high-affinity natural ligand l-fucose (8) for both lectin types (IC₅₀[LecB_PAO1] 2.74 μM; (30) IC₅₀[LecB_PA14] 0.91 μM). (20) On the basis of the four reference pairs analyzed, i.e., mannoside versus C-glycoside 2a/6a, 2b/6b, 3a/7a, and 3b/7b, which differ only at the anomeric position of mannose, a strong increase in affinity toward LecB of up to 16-fold was identified for the C-glycosides. In general, the introduction of amide and sulfonamide substituents always improved affinity for LecB compared to that of unsubstituted reference hybrid-structure 5. (31) Importantly, for the first time the two clinically relevant variants (20) of LecB were analyzed with glycomimetic inhibitors, which showed potent inhibition for both strain types.

The thermodynamics of binding of derivatives 6a and 7a,b to both LecB variants was then studied by isothermal titration calorimetry (Figure 3, Table S1–3). All ligands showed K_d values in the low micro- to nanomolar range and a 1:1 binding stoichiometry, confirming the IC₅₀ data obtained. Notably, compared to carbocyclic derivatives 6a and 7a (ΔH = −37.8 to −31.4 kJ/mol), thiophene-containing ligand 7b showed an enhanced enthalpy driven binding (ΔH = −50.0 and −48.1 kJ/mol), which was partially compensated by disfavored entropic contributions (−TΔS = 13.4 and 12.7 kJ/mol). Thus, the introduced substituents also significantly enhanced binding enthalpy compared to the unsubstituted congener 5 (ΔH = −27.5 kJ/mol). An increased enthalpic contribution to binding was previously introduced as a benchmark parameter assigned to a higher degree of target selectivity due to a better quality of the drug-target interaction. (36) Furthermore, physicochemical metrics such as ligand efficiency and related ones, thought to gauge the quality of a drug-target interaction, (37) are promising. Importantly, the most potent compound thiophene, 7b, displayed enthalphy-driven nanomolar affinities toward both LecB variants.

Figure 3. Isothermal titration microcalorimetry of LecB_PAO1 and LecB_PA14 with ligands 6a and 7a,b. Means and standard deviations were determined from a minimum of three independent titrations. One representative titration graph is depicted for LecB_PAO1 only.

We have previously detected a strong increase of the receptor half-lives for inhibitors 2a,b and 3a, compared to that of to natural carbohydrate ligand methyl α-d-mannoside (1). (32) The role of ligand–receptor binding kinetics is becoming increasingly significant for drug discovery and translation of in vitro binding kinetics into cellular or in vivo effects was discussed. (38) For the in vivo efficacy of a drug molecule, prolonged receptor residence times are of importance. Ligand–receptor binding kinetics of C-glycosides 6a and 7a were studied by surface plasmon resonance (SPR) with surface-bound LecB_PAO1 (Figure 4). With its affinity of 330 nM obtained from SPR, the hybrid-type structure 7a is a 140-fold more potent ligand of LecB than its carbohydrate ligand 1 and is 10-fold more potent than the natural high affinity ligand l-fucose (8) (see ITC experiments; K_d for 8 2.9 μM). (39) Most importantly, a further increase of the drug-receptor half-lives for the C-glycosides 6a and 7a over those of their mannose-based analogs 2a (t_1/2 6.3 min and K_d by SPR 18.1 μM) and 3a (t_1/2 18.6 min and K_d by SPR 1.12 μM) (32) was observed. Receptor residence times of 28 min for 6a and 7a resulted from very slow off rates of 4.1 × 10^–4 s^–1 of these C-glycosides. Thus, these glycomimetics form a 38-fold more stable complex with the protein, compared to the natural ligand methyl α-d-mannoside (1, t_1/2 0.75 min, K_d by SPR 47.5 μM). (32) These data validated the results from the competitive binding assay and ITC and further stress the impact of the additional equatorial methyl group in our glycomimetics.

Figure 4. Surface plasmon resonance (SPR) analysis of direct binding of 6a or 7a to immobilized LecB_PAO1. Experimental data in black; calculated fits using a 1:1 binding model in red.

Structure of LecB in Complex with Hybrid Inhibitors

To study the binding mode of the hybrid-type LecB inhibitors, we performed crystallization of these ligands in complex with the lectin. Crystals of two ligands 7a and 7b in complex with LecB_PA14 were obtained by the hanging drop cocrystallization method and the structures were solved to 1.65 Å resolution (Figure 5, Table S4). In both complexes, the carbohydrate-binding sites were occupied with the respective inhibitors except one site in the LecB/7a complex.

Figure 5. Crystal structures of LecB_PA14 with C-glycoside ligands: (A) Complex with trimethylphenyl sulfonamide 7a (1.65 Å resolution), (B) Complex with thiophene 7b (1.65 Å resolution); For the ligands the 2F_obs – F_calc electron density is displayed at 1 σ. Ligands and amino acids of the carbohydrate recognition domain (CRD) are depicted as sticks colored by elements (C: gray, N: blue, O: red, S: yellow); protein surface in transparent blue and two Ca²⁺-ions in the binding sites are shown as green spheres.

Surprisingly, a slightly different binding mode for methyl containing analog 7a was observed in contrast to the previously reported structure (30) of its mannose analog 3a (Figures 5A, S1, and S3). While the carbohydrate-derived ring is in an identical position, the aromatic moiety attached to the exocyclic CH₂ is rotated and located in a cleft of the protein surface that is enclosed by two neighboring loops (S68-D75 and E95-D104) of the lectin. The methyl group attached to the aromatic ring interacts with hydrophobic patches formed by V69 and a CH₂ group of D96; one sulfonamide oxygen of the ligand establishes a hydrogen bond with the backbone amide of G24. In contrast with mannoside 3a, a lipophilic contact between the protein surface (G24, V69) and the mesitylene-ring was observed together with a hydrogen bond of its sulfonamide nitrogen with the side chain of D96 (Figure S1). The latter interaction results from a conformational change of the C5–C6 bond in the carbohydrate from a gt-orientation (N gauche to O5 and trans to O4), as present in the complex of LecB with d-mannose (PDB code for LecB_PAO11OUR (40) or LecB_PA145A6Y), (20) to a tg-conformation in the mannoside 3a enabling this observed NH-D96 hydrogen bond. A binding mode comparable to that in C-glycoside 7a was detected for its analog thiophene 7b (Figures 5B, S1, and S3). In the structure of mannoside 3a/LecB, a crystal-lattice-dependent π-stacking of the aromatic moiety with a second ligand from a neighboring LecB tetramer is observed with all ligands in this structure showing the tg-conformation (Figure S2). Therefore, we consider the binding modes reported here with the gt-orientation displayed by C-glycosides 7a and 7b as the relevant ones, where crystal induced contacts are absent. In addition to the sulfonamide substituent interactions, a lipophilic contact of the introduced C-glycosidic methyl group with the protein residues T45 and A23 is observed in both analyzed complexes of hybrid-type ligands providing a molecular basis for the measured affinity enhancement.

Low Molecular Weight Glycomimetics Are Potent Inhibitors of Bacterial Biofilm Formation

We then tested the most promising compounds in a P. aeruginosa biofilm assay. Most published biofilm assays require staining and washing steps which impact the reproducibility in our hands. Therefore, we generated genetically modified P. aeruginosa which constitutively and intracellulary expresses the red-fluorescent protein mCherry from the pMP7605 plasmid. (41) Because fluorescence intensity directly correlated with cell density (Figure S4), mCherry-derived fluorescence was exploited as an internal stain for bacterial visualization and quantification of biofilm formation by confocal laser scanning fluorescence microscopy. By eliminating washing and staining steps, this method improved reproducibility through in situ imaging. A set of selected compounds comprising natural carbohydrates (mannoside 1, fucoside 11), glycomimetic mannose-derivatives (2a, 3a), and C-glycosides (6a, 7a,b) was selected for assessment of their potential to inhibit biofilm formation by P. aeruginosa.

To exclude any unwanted antibiotic effect on biofilm formation, the total fluorescence intensity was recorded after bacterial growth in the presence of compounds for 23 h. Bacteria grew to the same density as the DMSO control under each condition tested (Figure S5). Thus, these results clearly indicated the absence of bactericidal or bacteriostatic effects for the tested compounds.

For inhibition of biofilm formation, culture medium was inoculated with P. aeruginosa PA14/mCherry in the presence of 100 μM compounds, and culture and biofilm growth was allowed for 48 h, when biofilm mass was quantified using a confocal fluorescence microscope (Figure 6). All analyzed compounds reduced biofilm formation by P. aeruginosa. Remarkably, the natural carbohydrate ligands methyl α-d-mannoside (1) and methyl α-l-fucoside (11) showed only a small reduction in biofilm mass which was statistically insignificant. In contrast, all glycomimetics tested showed a significant reduction of biofilm formation with novel sulfonamide C-glycosides 7 as most potent compounds reaching approximately 80–90% of inhibition. Mannose analogs 2a and 3a also inhibited with a reduced potency the formation of biofilms with 54 and 66% observed reduction, respectively.

Figure 6. Inhibition of biofilm formation by *P. aeruginosa* after 48 h growth in the presence of compounds 1, 2a, 3a, 6a, 7a,b, or methyl α-l-fucoside (11) at 100 μM. DMSO in absence of compounds was used as control. (A) Quantification of biofilm biomass. Averages and standard deviations of biofilm formation from three independent replicates. Statistical significance was calculated using the students t test. (B) Raw data of confocal fluorescence microscopy 3D images show one representative z-stack per condition.

Compound Selectivity for LecB over Host Lectins

Carbohydrate-binding proteins are ubiquitous and many play important roles in host innate immune defense. (42) Thus, inhibitors of LecB carrying natural carbohydrate epitopes may also result in unspecific inhibition of human pathogen-recognition receptors. Langerin is a C-type lectin expressed in human Langerhans cells and CD103⁺ dermal dendritic cells and recognizes predominantly d-mannose but also l-fucose, N-acetyl-d-glucosamine and sulfated carbohydrates. (43, 44) As the C-glycosides presented here contain structural elements of d-mannose, l-fucose, and sulfonamide moieties, they are potential ligands of Langerin. Therefore, this protein was chosen as one representative host lectin to study compound specificity. In order to assess the compounds’ selectivity, 6a, 7a, and its natural ligand l-fucose (8) were analyzed for Langerin binding in a competitive ¹⁹F R₂-filtered NMR assay (44) (Figure 7). Natural ligand 8 was recognized by Langerin with slightly decreased affinity (K_i of 7.2 mM) compared to d-mannose (K_i 4.5 mM (44)); C-glycosides 6a and 7a also showed moderate binding to this human protein in the millimolar range. However, both compounds displayed a 1000-fold stronger affinity for LecB and are therefore considered selective for LecB over the host lectin Langerin.

Figure 7. Binding of C-glycosides 6a and 7a and l-fucose (8) to Langerin as determined in a ¹⁹F R₂-filtered NMR competitive binding assay.

In addition to the direct binding to one host lectin, a more global effect of the glycomimetics on immune homeostasis was investigated. Primary murine spleen cells were stimulated via different pathways using bacterial lipopolysaccharide (LPS), fungal mannan, or a combination of phorbol 12-myristate 13-acetate (PMA) and ionomycin, and the immune response to these stimuli in presence and absence of compounds 1, 2a, 3a, 6a, 7a, or 11 was monitored by quantifying TNF-α release (Figure S6). No effect of mannoside 1 on the TNF-α levels was observed in comparison to matched DMSO concentrations (Figure S6A), while cinnamide 2a showed a decrease in TNF-α levels (Figure S6B). An immune suppressive effect was exclusively observed for cinnamide 2a at the highest concentration tested (1 mM), while all other tested compounds, 3a, 6a, 7a, and 11, showed no effect on the immune response (Figure S6C). The immune-suppressive effect of 2a is an interesting observation; however, the high concentration needed probably impedes further development toward this effect. Excluding mannose-derived cinnamide 2a, none of the other compounds tested showed any effect on the immune response and provided further supporting evidence for compound selectivity.

In Vitro Metabolic Stability and Toxicity

To achieve effective compound concentrations in vivo, efficient compound uptake and stability toward host metabolism are crucial properties. Compound solubility is not considered problematic for these well-soluble carbohydrate derivatives. Calculated compound lipophilicity (clogP) showed reasonable values (−1.25 < clogP < −0.13) for all compounds selected for early pharmacokinetics and toxicity studies, i.e., sulfonamide C-glycosides 7a,b derivatives and two mannose-based compounds 3a and 3b (Table S5).

In vitro metabolic stability of the LecB inhibitors was studied against mouse and human liver microsomes and murine plasma (Tables S6 and S7, Figure S7). The data obtained revealed a low intrinsic clearance (CL_int) by mouse and human liver microsomes for all tested compounds. The metabolic stability of the C-glycoside 7a (CL_int 12 μL/min/mg protein) toward mouse microsomes was slightly improved compared to its O-glycoside analog 3a (CL_int 19 μL/min/mg protein) supporting the initial rational design of these C-glycosides. In the second group, the stabilities of mannoside 3b and C-glycoside 7b could not be differentiated since both compounds showed highest stability in this assay. Remarkably, all tested compounds showed very low clearance by human liver microsomes, where all substances fell into the highest stability category of this assay. Stability in murine plasma was assessed, and while the positive controls procaine and propoxycaine were rapidly degraded, all analyzed LecB ligands were as stable as the negative control procainamide and were not significantly degraded during the incubation period of 2 h. Therefore, all compounds tested can also be considered stable in murine plasma.

In order to assess a first safety window, toxicity of compounds was studied in vitro using an immortalized human hepatocyte cell line (Hep G2 cells, Figure S8). No toxicity was detected for all compounds tested up to a concentration of 100 μM.

In Vivo Pharmacokinetics of C-Glycosides

The two candidates from sulfonamides 7 with potent antibiofilm effects and favorable ADME properties, i.e., 7a and 7b, were chosen for an in vivo pharmacokinetics study in mice. Male CD-1 mice received a single dose of either 7a or 7b (10 mg/kg; n = 3) intravenously (i.v.) or perorally (p.o.). Plasma and urine levels of the parent compounds were monitored during 24 h (Figure 8) and pharmacokinetic parameters of 7a and 7b were assessed from plasma levels using a one-compartment model (Table S10). (45) In general, thiophene 7b had a superior plasma concentration over that of mesitylene 7a achieving a 2-fold higher calculated initial concentration (C₀ = 13.4 μM for 7a (i.v.); C₀ = 28.3 μM for 7b (i.v.)); thus, both candidates exceeded the anticipated necessary therapeutic levels (concentration > target K_d) with 10 mg/kg i.v.. Furthermore, 7b showed a slower elimination rate (k₁₀), a prolonged half-life (t_1/2) and mean residence time (MRT) and a low clearance (CL) compared to 7a. This resulted in higher compound levels in plasma at later time points. These features also contribute to an overall drug exposure in plasma for 7b at an AUC of 7.4 μg·h/mL using the intravenous administration route, which was around 4-fold higher than for 7a. The p.o. administration of 7a resulted in a bioavailability of nearly 100%, whereas for 7b it was only around 33.5%. However, the p.o. route of administration of 7b still resulted in a higher AUC than that for 7a in the i.v. route. In addition, the p.o. route of 7b showed similar elimination curves as detected for the i.v. route. In general, both compounds 7a and 7b were absorbed rapidly (t_max: 0.26–0.51 h for 7a and 7b, p.o. route), which might be a consequence of the sugar-like structural elements inherited by both compounds facilitating the crossing of barriers. As unmetabolized 7b was detected in urine at high concentrations (Figure 8), its elimination from the systemic circulation likely proceeds via renal clearance rather than liver detoxification. The kidneys did not act as a reservoir, and thiophene 7b did not accumulate in this organ, since its concentration in kidney 24 h post i.v. dosing was below 6 nM. Taken together, both compounds showed good oral bioavailability and thiophene 7b showed a superior pharmacokinetic profile compared to 7a with high compound levels, both in plasma and urine.

Figure 8. *In vivo* pharmacokinetics of 7a and 7b in CD-1 mice. Mean profile (±) SD of plasma (left column) and urine (right column) concentration in mouse versus time after i.v. (top) or p.o. (bottom) administration of compound 7a or 7b in a single dosing experiment (10 mg/kg, n = 3). Dashed line represents the *in vitro* IC₅₀ range for both candidates with LecB_PA14.

Conclusions

Click to copy section linkSection link copied!

We have developed the first low molecular weight LecB inhibitors that potently inhibited biofilm formation by P. aeruginosa, one of the prime mechanisms of antimicrobial resistance of this pathogen of outstanding medical importance. The synthesized glycomimetic C-glycosides showed inhibitory affinities as low as nanomolar for LecB variants from two strains representative for the entire set of clinical isolates. (20) Thus, it can be expected that the devised glycomimetics will be active against LecB-mediated biofilm formation in a broad spectrum of clinical isolates.

The compounds showed excellent biophysical properties with receptor residence times as high as 28 min and fully enthalpy driven binding to the target lectin, which are two important benchmarks for early drug development. In contrast to the ineffective natural carbohydrate ligands, our glycomimetics were potent inhibitors of biofilm formation without affecting bacterial viability. Thus, development of resistances toward these pure antibiofilm substances is likely to be reduced, in contrast to this well-known problem for traditional antibiotics. (12, 46)

Target selectivity was then studied in binding experiments toward Langerin, a human fucose- and mannose-binding C-type lectin, as well as in a more global approach in immune-stimulated primary murine spleen cells, and no impeding effects of the glycomimetics could be observed. All compounds tested showed absence of toxicity. Furthermore, a set of in vitro ADME experiments revealed good metabolic stability of the compounds against liver microsomes and murine and human plasma as well as a balanced lipophilicity that is important for oral availability.

In a first in vivo experiment, the pharmacokinetics of 7a and 7b in mice revealed oral bioavailability with high plasma concentrations and a subsequent primary excretion route via the kidneys. Consequently, high compound concentrations in urine were observed, suggesting but not limiting to an application in a P. aeruginosa urinary tract infection (UTI) model. It will now be of interest to evaluate the compounds’ anti-infective potential in a monotherapy treatment against biofilm-associated infections and their synergy together with antibiotics for efficient eradication of bacteria outside the biofilm.

Supporting Information

Click to copy section linkSection link copied!

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/jacs.7b11133.

Experimental details and ¹H and ¹³C spectra of new compounds; ITC titration data; X-ray data collection and refinement statistics; crystal structures showing ligand alignment and crystal packing effects; correlation of fluorescence intensities with cfu and OD600 measurements; effects of compounds on total fluorescence intensities; calculated lipophilicity of selected compounds; analysis of TNF-α concentration after stimulation of mouse spleen cells with and without test compounds; microsomal intrinsic clearance (CL_int) of 3a, 3b, and C-glycosides 7 in mouse and human liver microsomes; stability of LecB ligands in mouse plasma; m/z search window for plasma stability assay; toxicity of LecB ligands to human liver Hep G2 cells; accuracy, quantification limits, and lower limit of qualification for 7a and 7b in plasma, urine, and kidney matrix; mass spectrometric conditions used for quantification and qualification of 7a, 7b, and the internal standard glipizide; PK parameters of 7a and 7b in mice (PDF)

ja7b11133_si_001.pdf (4.35 MB)

Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Author Information

Click to copy section linkSection link copied!

Corresponding Author
- Alexander Titz - Chemical Biology of Carbohydrates and ‡Drug Design and Development, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), D-66123 Saarbrücken, Germany; Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany; Department of Pharmacy, Saarland University, D-66123 Saarbrücken, Germany; http://orcid.org/0000-0001-7408-5084; Email: [email protected]
Authors
- Roman Sommer - Chemical Biology of Carbohydrates and ‡Drug Design and Development, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), D-66123 Saarbrücken, Germany; Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Stefanie Wagner - Chemical Biology of Carbohydrates and ‡Drug Design and Development, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), D-66123 Saarbrücken, Germany; Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Katharina Rox - Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Annabelle Varrot - Univ. Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales (CERMAV), 38000 Grenoble, France
- Dirk Hauck - Chemical Biology of Carbohydrates and ‡Drug Design and Development, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), D-66123 Saarbrücken, Germany; Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Eike-Christian Wamhoff - Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, D-14424 Potsdam, Germany; Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin, D-14195 Berlin, Germany
- Janine Schreiber - Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Thomas Ryckmans - Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, CH-4070 Basel, Switzerland
- Thomas Brunner - Biochemical Pharmacology, Department of Biology, University of Konstanz, D-78457 Konstanz, Germany
- Christoph Rademacher - Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, D-14424 Potsdam, Germany; Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin, D-14195 Berlin, Germany; http://orcid.org/0000-0001-7082-7239
- Rolf W. Hartmann - Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany; Department of Pharmacy, Saarland University, D-66123 Saarbrücken, Germany; http://orcid.org/0000-0002-5871-5231
- Mark Brönstrup - Deutsches Zentrum für Infektionsforschung (DZIF), Standort Hannover-Braunschweig, Germany
- Anne Imberty - Univ. Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales (CERMAV), 38000 Grenoble, France
Notes
The authors declare no competing financial interest.

Acknowledgment

Click to copy section linkSection link copied!

We are grateful to Sarah Henrikus and Shelby Newsad for chemistry support and to Dr. Michael Hoffmann for HRMS measurements (all HIPS Saarbrücken). We acknowledge technical assistance from Tatjana Arnold (HZI, Braunschweig) and Astrid Glöckner (Konstanz University); Dr. Aymeric Audfray (CERMAV Grenoble) is acknowledged for instructions to SPR. We thank Dr. Josef Zapp (Saarland University) for performing NMR measurements. Crystal data collection was performed at the European Synchrotron Radiation Facility, Grenoble, France, and we are grateful for access and technical support to beamline FIP-BM30A and ID29. A.I. and A.V. acknowledge support from the ANR projects Glyco@Alps (ANR-15-IDEX-02) and Labex ARCANE (ANR-11-LABX-003). We thank the Helmholtz Association (grant no VH-NG-934, to A.T.), EU COST action BM1003 (to R.S.), DAAD RISE program (to S. Newsad and R.S.) and the Deutsche Forschungsgemeinschaft (to A.T., grant no. Ti756/2-1, to C.R., grant no. RA1944/2-1) for financial support.

References

Click to copy section linkSection link copied!

This article references 46 other publications.

1
Nagao, M.; Iinuma, Y.; Igawa, J.; Saito, T.; Yamashita, K.; Kondo, T.; Matsushima, A.; Takakura, S.; Takaori-Kondo, A.; Ichiyama, S. J. Hosp Infect 2011, 79, 49– 53 DOI: 10.1016/j.jhin.2011.04.018
Google Scholar
1
Control of an outbreak of carbapenem-resistant Pseudomonas aeruginosa in a haemato-oncology unit
Nagao M; Iinuma Y; Igawa J; Saito T; Yamashita K; Kondo T; Matsushima A; Takakura S; Takaori-Kondo A; Ichiyama S
The Journal of hospital infection (2011), 79 (1), 49-53 ISSN:.
An outbreak of a multidrug-resistant Pseudomonas aeruginosa producing metallo-β-lactamase (MBLPA) in a haemato-oncology unit was controlled using multidisciplinary interventions. The present study assesses the effects of these interventions by active surveillance of the incidence of MBLPA infection at the 1,240-bed tertiary care Kyoto University Hospital in Kyoto, Japan. Infection control strategies in 2004 included strengthening contact precautions, analysis of risk factors for MBLPA infection and cessation of urine collection. However, new MBLPA infections were identified in 2006, which prompted enhanced environmental cleaning, routine active surveillance, and restricting carbapenem usage. Between 2004 and 2010, 17 patients in the unit became infected with indistinguishable MBLPA strains. The final five infected patients were found by routine active surveillance, but horizontal transmission was undetectable. The MBLPA outbreak in the haemato-oncology unit was finally contained in 2008.
2
Rice, L. B. J. Infect. Dis. 2008, 197, 1079– 1081 DOI: 10.1086/533452
Google Scholar
2
Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE
Rice Louis B
The Journal of infectious diseases (2008), 197 (8), 1079-81 ISSN:0022-1899.
There is no expanded citation for this reference.
3
Tsutsui, A.; Suzuki, S.; Yamane, K.; Matsui, M.; Konda, T.; Marui, E.; Takahashi, K.; Arakawa, Y. J. Hosp. Infect. 2011, 78, 317– 322 DOI: 10.1016/j.jhin.2011.04.013
Google Scholar
3
Genotypes and infection sites in an outbreak of multidrug-resistant Pseudomonas aeruginosa
Tsutsui A; Suzuki S; Yamane K; Matsui M; Konda T; Marui E; Takahashi K; Arakawa Y
The Journal of hospital infection (2011), 78 (4), 317-22 ISSN:.
An outbreak of multidrug-resistant (MDR) Pseudomonas aeruginosa occurred in an acute care hospital in Japan, which lasted for more than three years. During January 2006 to June 2009, 59 hospitalised patients with MDR P. aeruginosa were mainly detected by urine culture in the first half, whereas isolation from respiratory tract samples became dominant in the latter half of the outbreak. Non-duplicate MDR P. aeruginosa isolates were available from 51 patients and all isolates were positive for bla(VIM-2). Pulsed-field gel electrophoresis (PFGE) analysis categorised the isolates into three major clusters; types A, B and C with eight, 19 and 21 isolates, respectively. The outbreak started with patients harbouring PFGE type A strains, followed by type B, and type C strains. Multivariate analysis demonstrated that patients with PFGE type C strains were more likely to be detected by respiratory tract samples (odds ratio: 11.87; 95% confidence interval: 1.21-116.86). Improved aseptic urethral catheter care controlled PFGE type A and type B strains and improvement in respiratory care procedures finally contained the transmission of PFGE type C strains.
4
World Health Organization. WHO publishes list of bacteria for which new antibiotics are urgently needed. http://www.who.int/mediacentre/news/releases/2017/bacteria-antibiotics-needed/en/ (accessed May 2017).
Google Scholar
There is no corresponding record for this reference.
5
Hauser, A. R.; Rello, J., Eds. Severe Infections Caused by Pseudomonas aeruginosa; Perspectives on Critical Care Infectious Diseases series; Kluwer Academic Publishers Group: Boston, MA, 2003.
Google Scholar
There is no corresponding record for this reference.
6
Serra, R.; Grande, R.; Butrico, L.; Rossi, A.; Settimio, U. F.; Caroleo, B.; Amato, B.; Gallelli, L.; de Franciscis, S. Expert Rev. Anti-Infect. Ther. 2015, 13, 605– 613 DOI: 10.1586/14787210.2015.1023291
Google Scholar
6
Chronic wound infections: the role of Pseudomonas aeruginosa and Staphylococcus aureus
Serra, Raffaele; Grande, Raffaele; Butrico, Lucia; Rossi, Alessio; Settimio, Ugo Francesco; Caroleo, Benedetto; Amato, Bruno; Gallelli, Luca; de Franciscis, Stefano
Expert Review of Anti-Infective Therapy (2015), 13 (5), 605-613CODEN: ERATCK; ISSN:1478-7210. (Informa Healthcare)
A review. Chronic leg ulcers affect 1-2% of the general population and are related to increased morbidity and health costs. Staphylococcus aureus and Pseudomonas aeruginosa are the most common bacteria isolated from chronic wounds. They can express virulence factors and surface proteins affecting wound healing. The co-infection of S. aureus and P. aeruginosa is more virulent than single infection. In particular, S. aureus and P. aeruginosa have both intrinsic and acquired antibiotic resistance, making clin. management of infection a real challenge, particularly in patients with comorbidity. Therefore, a correct and prompt diagnosis of chronic wound infection requires a detailed knowledge of skin bacterial flora. This is a necessary prerequisite for tailored pharmacol. treatment, improving symptoms, and reducing side effects and antibiotic resistance.
7
Mittal, R.; Aggarwal, S.; Sharma, S.; Chhibber, S.; Harjai, K. J. Infect. Public Health 2009, 2, 101– 11 DOI: 10.1016/j.jiph.2009.08.003
Google Scholar
7
Urinary tract infections caused by Pseudomonas aeruginosa: a minireview
Mittal Rahul; Aggarwal Sudhir; Sharma Saroj; Chhibber Sanjay; Harjai Kusum
Journal of infection and public health (2009), 2 (3), 101-11 ISSN:.
Urinary tract infections (UTIs) are a serious health problem affecting millions of people each year. Infections of the urinary tract are the second most common type of infection in the body. Catheterization of the urinary tract is the most common factor, which predisposes the host to these infections. Catheter-associated UTI (CAUTI) is responsible for 40% of nosocomial infections, making it the most common cause of nosocomial infection. CAUTI accounts for more than 1 million cases in hospitals and nursing homes annually and often involve uropathogens other than Escherichia coli. While the epidemiology and pathogenic mechanisms of uropathogenic Escherichia coli have been extensively studied, little is known about the pathogenesis of UTIs caused by other organisms like Pseudomonas aeruginosa. Scanty available information regarding pathogenesis of UTIs caused by P. aeruginosa is an important bottleneck in developing effective preventive approaches. The aim of this review is to summarize some of the advances made in the field of P. aeruginosa induced UTIs and draws attention of the workers that more basic research at the level of pathogenesis is needed so that novel strategies can be designed.
8
Poole, K. Front. Microbiol. 2011, 2, 65 DOI: 10.3389/fmicb.2011.00065
Google Scholar
8
Pseudomonas aeruginosa: resistance to the max
Poole, Keith
Frontiers in Cellular and Infection Microbiology (2011), 2 (April), 65CODEN: FCIMAB ISSN:. (Frontiers Media S.A.)
A review. Pseudomonas aeruginosa is intrinsically resistant to a variety of antimicrobials and can develop resistance during anti-pseudomonal chemotherapy both of which compromise treatment of infections caused by this organism. Resistance to multiple classes of antimicrobials (multidrug resistance) in particular is increasingly common in P. aeruginosa, with a no. of reports of pan-resistant isolates treatable with a single agent, colistin. Acquired resistance in this organism is multifactorial and attributable to chromosomal mutations and the acquisition of resistance genes via horizontal gene transfer. Mutational changes impacting resistance include upregulation of multidrug efflux systems to promote antimicrobial expulsion, derepression of ampC, AmpC alterations that expand the enzyme's substrate specificity (i.e., extended-spectrum AmpC), alterations to outer membrane permeability to limit antimicrobial entry and alterations to antimicrobial targets. Acquired mechanisms contributing to resistance in P. aeruginosa include β-lactamases, notably the extended-spectrum β-lactamases and the carbapenemases that hydrolyze most β-lactams, aminoglycoside-modifying enzymes, and 16S rRNA methylases that provide high-level pan-aminoglycoside resistance. The organism's propensity to grow in vivo as antimicrobial-tolerant biofilms and the occurrence of hypermutator strains that yield antimicrobial resistant mutants at higher frequency also compromise anti-pseudomonal chemotherapy. With limited therapeutic options and increasing resistance will the untreatable P. aeruginosa infection soon be upon us.
9
Flemming, H.-C.; Wingender, J. Nat. Rev. Microbiol. 2010, 8, 623– 633 DOI: 10.1038/nrmicro2415
Google Scholar
9
The biofilm matrix
Flemming, Hans-Curt; Wingender, Jost
Nature Reviews Microbiology (2010), 8 (9), 623-633CODEN: NRMACK; ISSN:1740-1526. (Nature Publishing Group)
A review. The microorganisms in biofilms live in a self-produced matrix of hydrated extracellular polymeric substances (EPS) that form their immediate environment. EPS are mainly polysaccharides, proteins, nucleic acids and lipids; they provide the mech. stability of biofilms, mediate their adhesion to surfaces and form a cohesive, three-dimensional polymer network that interconnects and transiently immobilizes biofilm cells. In addn., the biofilm matrix acts as an external digestive system by keeping extracellular enzymes close to the cells, enabling them to metabolize dissolved, colloidal and solid biopolymers. Here we describe the functions, properties and constituents of the EPS matrix that make biofilms the most successful forms of life on earth.
10
Davies, D. Nat. Rev. Drug Discovery 2003, 2, 114– 122 DOI: 10.1038/nrd1008
Google Scholar
There is no corresponding record for this reference.
11
Sommer, R.; Joachim, I.; Wagner, S.; Titz, A. Chimia 2013, 67, 286– 290 DOI: 10.2533/chimia.2013.286
Google Scholar
There is no corresponding record for this reference.
12
Wagner, S.; Sommer, R.; Hinsberger, S.; Lu, C.; Hartmann, R. W.; Empting, M.; Titz, A. J. Med. Chem. 2016, 59, 5929– 5969 DOI: 10.1021/acs.jmedchem.5b01698
Google Scholar
There is no corresponding record for this reference.
13
Winzer, K.; Falconer, C.; Garber, N. C.; Diggle, S. P.; Camara, M.; Williams, P. J. Bacteriol. 2000, 182, 6401– 6411 DOI: 10.1128/JB.182.22.6401-6411.2000
Google Scholar
13
The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS
Winzer, Klaus; Falconer, Colin; Garber, Nachman C.; Diggle, Stephen P.; Camara, Miguel; Williams, Paul
Journal of Bacteriology (2000), 182 (22), 6401-6411CODEN: JOBAAY; ISSN:0021-9193. (American Society for Microbiology)
In Pseudomonas aeruginosa, many exoproduct virulence determinants are regulated via a hierarchical quorum-sensing cascade involving the transcriptional regulators LasR and RhlR and their cognate activators, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL). In this paper, we demonstrate that the cytotoxic lectins PA-IL and PA-IIL are regulated via quorum sensing. Using immunoblot anal., the prodn. of both lectins was found to be directly dependent on the rhl locus while, in a lasR mutant, the onset of lectin synthesis was delayed but not abolished. The PA-IL structural gene, lecA, was cloned and sequenced. Transcript anal. indicated a monocistronic organization with a transcriptional start site 70 bp upstream of the lecA translational start codon. A lux box-type element together with RpoS (σS) consensus sequences was identified upstream of the putative promoter region. In Escherichia coli, expression of a lecA::lux reporter fusion was activated by RhlR/C4-HSL, but not by LasR/3O-C12-HSL, confirming direct regulation by RhlR/C4-HSL. Similarly, in P. aeruginosa PAO1, the expression of a chromosomal lecA::lux fusion was enhanced but not advanced by the addn. of exogenous C4-HSL but not 3O-C12-HSL. Furthermore, mutation of rpoS abolished lectin synthesis in P. aeruginosa, demonstrating that both RpoS and RhlR/C4-HSL are required. Although the C4-HSL-dependent expression of the lecA::lux reporter in E. coli could be inhibited by the presence of 3O-C12-HSL, this did not occur in P. aeruginosa. This suggests that, in the homologous genetic background, 3O-C12-HSL does not function as a posttranslational regulator of the RhlR/C4-HSL-dependent activation of lecA expression.
14
Diggle, S. P.; Stacey, R. E.; Dodd, C.; Cámara, M.; Williams, P.; Winzer, K. Environ. Microbiol. 2006, 8, 1095– 1104 DOI: 10.1111/j.1462-2920.2006.001001.x
Google Scholar
There is no corresponding record for this reference.
15
Tielker, D.; Hacker, S.; Loris, R.; Strathmann, M.; Wingender, J.; Wilhelm, S.; Rosenau, F.; Jaeger, K.-E. Microbiology 2005, 151, 1313– 1323 DOI: 10.1099/mic.0.27701-0
Google Scholar
There is no corresponding record for this reference.
16
Gilboa-Garber, N. Methods Enzymol. 1982, 83, 378– 385 DOI: 10.1016/0076-6879(82)83034-6
Google Scholar
There is no corresponding record for this reference.
17
Klockgether, J.; Cramer, N.; Wiehlmann, L.; Davenport, C. F.; Tümmler, B. Front. Microbiol. 2011, 2, 150 DOI: 10.3389/fmicb.2011.00150
Google Scholar
17
Pseudomonas aeruginosa genomic structure and diversity
Klockgether, Jens; Cramer, Nina; Wiehlmann, Lutz; Davenport, Colin F.; Tuemmler, Burkhard
Frontiers in Cellular and Infection Microbiology (2011), 2 (July), 150CODEN: FCIMAB ISSN:. (Frontiers Media S.A.)
A review. The Pseudomonas aeruginosa genome (G+C content 65-67%, size 5.5-7 Mbp) is made up of a single circular chromosome and a variable no. of plasmids. Sequencing of complete genomes or blocks of the accessory genome has revealed that the genome encodes a large repertoire of transporters, transcriptional regulators and two-component regulatory systems which reflects its metabolic diversity to utilize a broad range of nutrients. The conserved core component of the genome is largely collinear among P. aeruginosa strains and exhibits an interclonal sequence diversity of 0.5-0.7 %. Only a few loci of the core genome are subject to diversifying selection. Genome diversity is mainly caused by accessory DNA elements located in 79 regions of genome plasticity that are scattered around the genome and show an anomalous usage of mono- to tetradecanucleotides. Genomic islands of the pKLC102/PAGI-2 family that integrate into tRNALys or tRNAGly genes represent hotspots of inter- and intra-clonal genomic diversity. The individual islands differ in their repertoire of metabolic genes that make a large contribution to the pangenome. In order to unravel intraclonal diversity of P. aeruginosa, the genomes of two members of the PA14 clonal complex from diverse habitats and geog. origin were compared. The genome sequences differed by less than 0.01% from each other. 198 Of the 231 SNPs were non-randomly distributed in the genome. Non-synonymous SNPs were mainly found in an integrated Pf1-like phage and in genes involved in transcriptional regulation, membrane and extracellular constituents, transport and secretion. In summary, P. aeruginosa is endowed with a highly conserved core genome of low sequence diversity and a highly variable accessory genome that communicates with other pseudomonads and genera via horizontal gene transfer.
18
Dötsch, A.; Schniederjans, M.; Khaledi, A.; Hornischer, K.; Schulz, S.; Bielecka, A.; Eckweiler, D.; Pohl, S.; Häussler, S. mBio 2015, 6, e00749-15 DOI: 10.1128/mBio.00749-15
Google Scholar
There is no corresponding record for this reference.
19
Boukerb, A. M.; Decor, A.; Ribun, S.; Tabaroni, R.; Rousset, A.; Commin, L.; Buff, S.; Doléans-Jordheim, A.; Vidal, S.; Varrot, A.; Imberty, A.; Cournoyer, B. Front. Microbiol. 2016, 7, 811 DOI: 10.3389/fmicb.2016.00811
Google Scholar
19
Genomic Rearrangements and Functional Diversification of lecA and lecB Lectin-Coding Regions Impacting the Efficacy of Glycomimetics Directed against Pseudomonas aeruginosa
Boukerb Amine M; Ribun Sebastien; Doleans-Jordheim Anne; Cournoyer Benoit; Decor Aude; Tabaroni Rachel; Varrot Annabelle; Imberty Anne; Rousset Audric; Vidal Sebastien; Commin Loris; Buff Samuel
Frontiers in microbiology (2016), 7 (), 811 ISSN:1664-302X.
LecA and LecB tetrameric lectins take part in oligosaccharide-mediated adhesion-processes of Pseudomonas aeruginosa. Glycomimetics have been designed to block these interactions. The great versatility of P. aeruginosa suggests that the range of application of these glycomimetics could be restricted to genotypes with particular lectin types. The likelihood of having genomic and genetic changes impacting LecA and LecB interactions with glycomimetics such as galactosylated and fucosylated calix[4]arene was investigated over a collection of strains from the main clades of P. aeruginosa. Lectin types were defined, and their ligand specificities were inferred. These analyses showed a loss of lecA among the PA7 clade. Genomic changes impacting lec loci were thus assessed using strains of this clade, and by making comparisons with the PAO1 genome. The lecA regions were found challenged by phage attacks and PAGI-2 (genomic island) integrations. A prophage was linked to the loss of lecA. The lecB regions were found less impacted by such rearrangements but greater lecB than lecA genetic divergences were recorded. Sixteen combinations of LecA and LecB types were observed. Amino acid variations were mapped on PAO1 crystal structures. Most significant changes were observed on LecBPA7, and found close to the fucose binding site. Glycan array analyses were performed with purified LecBPA7. LecBPA7 was found less specific for fucosylated oligosaccharides than LecBPAO1, with a preference for H type 2 rather than type 1, and Lewis(a) rather than Lewis(x). Comparison of the crystal structures of LecBPA7 and LecBPAO1 in complex with Lewis(a) showed these changes in specificity to have resulted from a modification of the water network between the lectin, galactose and GlcNAc residues. Incidence of these modifications on the interactions with calix[4]arene glycomimetics at the cell level was investigated. An aggregation test was used to establish the efficacy of these ligands. Great variations in the responses were observed. Glycomimetics directed against LecB yielded the highest numbers of aggregates for strains from all clades. The use of a PAO1ΔlecB strain confirmed a role of LecB in this aggregation phenotype. Fucosylated calix[4]arene showed the greatest potential for a use in the prevention of P. aeruginosa infections.
20
Sommer, R.; Wagner, S.; Varrot, A.; Nycholat, C. M.; Khaledi, A.; Haussler, S.; Paulson, J. C.; Imberty, A.; Titz, A. Chem. Sci. 2016, 7, 4990– 5001 DOI: 10.1039/C6SC00696E
Google Scholar
20
The virulence factor LecB varies in clinical isolates: consequences for ligand binding and drug discovery
Sommer, Roman; Wagner, Stefanie; Varrot, Annabelle; Nycholat, Corwin M.; Khaledi, Ariane; Haeussler, Susanne; Paulson, James C.; Imberty, Anne; Titz, Alexander
Chemical Science (2016), 7 (8), 4990-5001CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
P. aeruginosa causes a substantial no. of nosocomial infections and is the leading cause of death of cystic fibrosis patients. This Gram-neg. bacterium is highly resistant against antibiotics and further protects itself by forming a biofilm. Moreover, a high genomic variability among clin. isolates complicates therapy. Its lectin LecB is a virulence factor and necessary for adhesion and biofilm formation. The authors analyzed the sequence of LecB variants in a library of clin. isolates and demonstrate that it can serve as a marker for strain family classification. LecB from the highly virulent model strain PA14 presents 13% sequence divergence with LecB from the well characterized PAO1 strain. These differences might result in differing ligand binding specificities and ultimately in reduced efficacy of drugs directed towards LecB. Despite several amino acid variations at the carbohydrate binding site, glycan array anal. showed a comparable binding pattern for both variants. A common high affinity ligand could be identified and after its chemoenzymic synthesis verified in a competitive binding assay: an N-glycan presenting two blood group O epitopes (H-type 2 antigen). Mol. modeling of the complex suggests a bivalent interaction of the ligand with the LecB tetramer by bridging two sep. binding sites. This binding rationalizes the strong avidity (35 nM) of LecBPA14 to this human fucosylated N-glycan. Biochem. evaluation of a panel of glycan ligands revealed that LecBPA14 demonstrated higher glycan affinity compared to LecBPAO1 including the extraordinarily potent affinity of 70 nM towards the monovalent human antigen Lewisa. The structural basis of this unusual high affinity ligand binding for lectins was rationalized by solving the protein crystal structures of LecBPA14 with several glycans.
21
Boukerb, A. M.; Rousset, A.; Galanos, N.; Méar, J.-B.; Thepaut, M.; Grandjean, T.; Gillon, E.; Cecioni, S.; Abderrahmen, C.; Faure, K.; Redelberger, D.; Kipnis, E.; Dessein, R.; Havet, S.; Darblade, B.; Matthews, S. E.; de Bentzmann, S.; Guéry, B.; Cournoyer, B.; Imberty, A.; Vidal, S. J. Med. Chem. 2014, 57, 10275– 10289 DOI: 10.1021/jm500038p
Google Scholar
There is no corresponding record for this reference.
22
Chemani, C.; Imberty, A.; de Bentzmann, S.; Pierre, M.; Wimmerová, M.; Guery, B. P.; Faure, K. Infect. Immun. 2009, 77, 2065– 2075 DOI: 10.1128/IAI.01204-08
Google Scholar
22
Role of LecA and LecB lectins in Pseudomonas aeruginosa-induced lung injury and effect of carbohydrate ligands
Chemani, Chanez; Imberty, Anne; de Bentzmann, Sophie; Pierre, Maud; Wimmerova, Michaela; Guery, Benoit P.; Faure, Karine
Infection and Immunity (2009), 77 (5), 2065-2075CODEN: INFIBR; ISSN:0019-9567. (American Society for Microbiology)
Pseudomonas aeruginosa is a frequently encountered pathogen that is involved in acute and chronic lung infections. Lectin-mediated bacterium-cell recognition and adhesion are crit. steps in initiating P. aeruginosa pathogenesis. This study was designed to evaluate the contributions of LecA and LecB to the pathogenesis of P. aeruginosa-mediated acute lung injury. Using an in vitro model with A549 cells and an exptl. in vivo murine model of acute lung injury, we compared the parental strain to lecA and lecB mutants. The effects of both LecA- and LecB-specific lectin-inhibiting carbohydrates (α-methyl-galactoside and α-methyl-fucoside, resp.) were evaluated. In vitro, the parental strain was assocd. with increased cytotoxicity and adhesion on A549 cells compared to the lecA and lecB mutants. In vivo, the P. aeruginosa-induced increase in alveolar barrier permeability was reduced with both mutants. The bacterial burden and dissemination were decreased for both mutants compared with the parental strain. Coadministration of specific lectin inhibitors markedly reduced lung injury and mortality. Our results demonstrate that there is a relationship between lectins and the pathogenicity of P. aeruginosa. Inhibition of the lectins by specific carbohydrates may provide new therapeutic perspectives.
23
Cott, C.; Thuenauer, R.; Landi, A.; Kühn, K.; Juillot, S.; Imberty, A.; Madl, J.; Eierhoff, T.; Römer, W. Biochim. Biophys. Acta, Mol. Cell Res. 2016, 1863, 1106– 1118 DOI: 10.1016/j.bbamcr.2016.02.004
Google Scholar
23
Pseudomonas aeruginosa lectin LecB inhibits tissue repair processes by triggering β-catenin degradation
Cott, Catherine; Thuenauer, Roland; Landi, Alessia; Kuehn, Katja; Juillot, Samuel; Imberty, Anne; Madl, Josef; Eierhoff, Thorsten; Roemer, Winfried
Biochimica et Biophysica Acta, Molecular Cell Research (2016), 1863 (6_Part_A), 1106-1118CODEN: BBAMCO; ISSN:0167-4889. (Elsevier B.V.)
Pseudomonas aeruginosa is an opportunistic pathogen that induces severe lung infections such as ventilator-assocd. pneumonia and acute lung injury. Under these conditions, the bacterium diminishes epithelial integrity and inhibits tissue repair mechanisms, leading to persistent infections. Understanding the involved bacterial virulence factors and their mode of action is essential for the development of new therapeutic approaches. In our study we discovered a so far unknown effect of the P. aeruginosa lectin LecB on host cell physiol. LecB alone was sufficient to attenuate migration and proliferation of human lung epithelial cells and to induce transcriptional activity of NF-κB. These effects are characteristic of impaired tissue repair. Moreover, we found a strong degrdn. of β-catenin, which was partially recovered by the proteasome inhibitor lactacystin. In addn., LecB induced loss of cell-cell contacts and reduced expression of the β-catenin targets c-myc and cyclin D1. Blocking of LecB binding to host cell plasma membrane receptors by sol. L-fucose prevented these changes in host cell behavior and signaling, and thereby provides a powerful strategy to suppress LecB function. Our findings suggest that P. aeruginosa employs LecB as a virulence factor to induce β-catenin degrdn., which then represses processes that are directly linked to tissue recovery.
24
Mitchell, E.; Houles, C.; Sudakevitz, D.; Wimmerova, M.; Gautier, C.; Pérez, S.; Wu, A. M.; Gilboa-Garber, N.; Imberty, A. Nat. Struct. Biol. 2002, 9, 918– 921 DOI: 10.1038/nsb865
Google Scholar
There is no corresponding record for this reference.
25
Hauber, H.-P.; Schulz, M.; Pforte, A.; Mack, D.; Zabel, P.; Schumacher, U. Int. J. Med. Sci. 2008, 5, 371– 376 DOI: 10.7150/ijms.5.371
Google Scholar
25
Inhalation with fucose and galactose for treatment of Pseudomonas aeruginosa in cystic fibrosis patients
Hauber, Hans-Peter; Schulz, Maria; Pforte, Almuth; Mack, Dietrich; Zabel, Peter; Schumacher, Udo
International Journal of Medical Sciences (2008), 5 (6), 371-376CODEN: IJMSGZ; ISSN:1449-1907. (Ivyspring International Publisher)
Background: Colonisation of cystic fibrosis (CF) lungs with Pseudomonas aeruginosa is facilitated by two lectins, which bind to the sugar coat of the surface lining epithelia and stop the cilia beating. Objectives: We hypothesized that P. aeruginosa lung infection should be cleared by inhalation of fucose and galactose, which compete for the sugar binding site of the two lectins and thus inhibit the binding of P. aeruginosa. Methods: 11 adult CF patients with chronic infection with P. aeruginosa were treated twice daily with inhalation of a fucose/galactose soln. for 21 days (4 patients only received inhalation, 7 patients received inhalation and i.v. antibiotics). Microbial counts of P. aeruginosa, lung function measurements, and inflammatory markers were detd. before and after treatment. Results: The sugar inhalation was well tolerated and no adverse side effects were obsd. Inhalation alone as well as combined therapy (inhalation and antibiotics) significantly decreased P. aeruginosa in sputum (P < 0.05). Both therapies also significantly reduced TNFα expression in sputum and peripheral blood cells (P < 0.05). No change in lung function measurements was obsd. Conclusions: Inhalation of simple sugars is a safe and effective measure to reduce the P. aeruginosa counts in CF patients. This may provide an alternative therapeutical approach to treat infection with P. aeruginosa.
26
Bucior, I.; Abbott, J.; Song, Y.; Matthay, M. A.; Engel, J. N. Am. J. Physiol Lung Cell Mol. Physiol 2013, 305, L352– 363 DOI: 10.1152/ajplung.00387.2012
Google Scholar
26
Sugar administration is an effective adjunctive therapy in the treatment of Pseudomonas aeruginosa pneumonia
Bucior, Iwona; Abbott, Jason; Song, Yuanlin; Matthay, Michael A.; Engel, Joanne N.
American Journal of Physiology (2013), 305 (3, Pt. 1), L352-L363CODEN: AJPHAP; ISSN:0002-9513. (American Physiological Society)
Treatment of acute and chronic pulmonary infections caused by opportunistic pathogen Pseudomonas aeruginosa is limited by the increasing frequency of multidrug bacterial resistance. Here, we describe a novel adjunctive therapy in which administration of a mix of simple sugars-mannose, fucose, and galactose-inhibits bacterial attachment, limits lung damage, and potentiates conventional antibiotic therapy. The sugar mixt. inhibits adhesion of nonmucoid and mucoid P. aeruginosa strains to bronchial epithelial cells in vitro. In a murine model of acute pneumonia, treatment with the sugar mixt. alone diminishes lung damage, bacterial dissemination to the subpleural alveoli, and neutrophil- and IL-8-driven inflammatory responses. Remarkably, the sugars act synergistically with anti-Pseudomonas antibiotics, including β-lactams and quinolones, to further reduce bacterial lung colonization and damage. To probe the mechanism, we examd. the effects of sugars in the presence or absence of antibiotics during growth in liq. culture and in an ex vivo infection model utilizing freshly dissected mouse tracheas and lungs. We demonstrate that the sugar mixt. induces rapid but reversible formation of bacterial clusters that exhibited enhanced susceptibility to antibiotics compared with individual bacteria. Our findings reveal that sugar inhalation, an inexpensive and safe therapeutic, could be used in combination with conventional antibiotic therapy to more effectively treat P. aeruginosa lung infections.
27
Cecioni, S.; Imberty, A.; Vidal, S. Chem. Rev. 2015, 115, 525– 561 DOI: 10.1021/cr500303t
Google Scholar
There is no corresponding record for this reference.
28
Bernardi, A.; Jiménez-Barbero, J.; Casnati, A.; De Castro, C.; Darbre, T.; Fieschi, F.; Finne, J.; Funken, H.; Jaeger, K.-E.; Lahmann, M.; Lindhorst, T. K.; Marradi, M.; Messner, P.; Molinaro, A.; Murphy, P. V.; Nativi, C.; Oscarson, S.; Penadés, S.; Peri, F.; Pieters, R. J.; Renaudet, O.; Reymond, J.-L.; Richichi, B.; Rojo, J.; Sansone, F.; Schäffer, C.; Turnbull, W. B.; Velasco-Torrijos, T.; Vidal, S.; Vincent, S.; Wennekes, T.; Zuilhof, H.; Imberty, A. Chem. Soc. Rev. 2013, 42, 4709– 4727 DOI: 10.1039/C2CS35408J
Google Scholar
There is no corresponding record for this reference.
29
Johansson, E. M. V.; Crusz, S. A.; Kolomiets, E.; Buts, L.; Kadam, R. U.; Cacciarini, M.; Bartels, K.-M.; Diggle, S. P.; Cámara, M.; Williams, P.; Loris, R.; Nativi, C.; Rosenau, F.; Jaeger, K.-E.; Darbre, T.; Reymond, J.-L. Chem. Biol. 2008, 15, 1249– 1257 DOI: 10.1016/j.chembiol.2008.10.009
Google Scholar
29
Inhibition and Dispersion of Pseudomonas aeruginosa Biofilms by Glycopeptide Dendrimers Targeting the Fucose-Specific Lectin LecB
Johansson, Emma M. V.; Crusz, Shanika A.; Kolomiets, Elena; Buts, Lieven; Kadam, Rameshwar U.; Cacciarini, Martina; Bartels, Kai-Malte; Diggle, Stephen P.; Camara, Miguel; Williams, Paul; Loris, Remy; Nativi, Cristina; Rosenau, Frank; Jaeger, Karl-Erich; Darbre, Tamis; Reymond, Jean-Louis
Chemistry & Biology (Cambridge, MA, United States) (2008), 15 (12), 1249-1257CODEN: CBOLE2; ISSN:1074-5521. (Cell Press)
The human pathogenic bacterium Pseudomonas aeruginosa produces a fucose-specific lectin, LecB, implicated in tissue attachment and the formation of biofilms. To investigate if LecB inhibition disrupts these processes, high-affinity ligands were obtained by screening two 15,536-member combinatorial libraries of multivalent fucosyl-peptide dendrimers. The most potent LecB-ligands identified were dendrimers FD2 (C-Fuc-LysProLeu)4(LysPheLysIle)2 LysHisIleNH2 (IC50 = 0.14 μM by ELLA) and PA8 (OFuc-LysAlaAsp)4(LysSerGlyAla)2 LysHisIleNH2 (IC50 = 0.11 μM by ELLA). Dendrimer FD2 led to complete inhibition of P. aeruginosa biofilm formation (IC50 ∼ 10 μM) and induced complete dispersion of established biofilms in the wild-type strain and in several clin. P. aeruginosa isolates. These expts. suggest that LecB inhibition by high-affinity multivalent ligands could represent a therapeutic approach against P. aeruginosa infections by inhibition of biofilm formation and dispersion of established biofilms.
30
Hauck, D.; Joachim, I.; Frommeyer, B.; Varrot, A.; Philipp, B.; Möller, H. M.; Imberty, A.; Exner, T. E.; Titz, A. ACS Chem. Biol. 2013, 8, 1775– 1784 DOI: 10.1021/cb400371r
Google Scholar
There is no corresponding record for this reference.
31
Sommer, R.; Exner, T. E.; Titz, A. PLoS One 2014, 9, e112822 DOI: 10.1371/journal.pone.0112822
Google Scholar
There is no corresponding record for this reference.
32
Sommer, R.; Hauck, D.; Varrot, A.; Wagner, S.; Audfray, A.; Prestel, A.; Möller, H. M.; Imberty, A.; Titz, A. ChemistryOpen 2015, 4, 756– 767 DOI: 10.1002/open.201500162
Google Scholar
There is no corresponding record for this reference.
33
Hofmann, A.; Sommer, R.; Hauck, D.; Stifel, J.; Göttker-Schnetmann, I.; Titz, A. Carbohydr. Res. 2015, 412, 34– 42 DOI: 10.1016/j.carres.2015.04.010
Google Scholar
There is no corresponding record for this reference.
34
Beshr, G.; Sommer, R.; Hauck, D.; Siebert, D. C. B.; Hofmann, A.; Imberty, A.; Titz, A. MedChemComm 2016, 7, 519– 530 DOI: 10.1039/C5MD00557D
Google Scholar
There is no corresponding record for this reference.
35
Sabin, C.; Mitchell, E. P.; Pokorná, M.; Gautier, C.; Utille, J.-P.; Wimmerová, M.; Imberty, A. FEBS Lett. 2006, 580, 982– 987 DOI: 10.1016/j.febslet.2006.01.030
Google Scholar
There is no corresponding record for this reference.
36
Tarcsay, A.; Keseru, G. M. Drug Discovery Today 2015, 20, 86– 94 DOI: 10.1016/j.drudis.2014.09.014
Google Scholar
36
Is there a link between selectivity and binding thermodynamics profiles?
Tarcsay, Akos; Keseru, Gyorgy M.
Drug Discovery Today (2015), 20 (1), 86-94CODEN: DDTOFS; ISSN:1359-6446. (Elsevier Ltd.)
A review. Thermodn. of ligand binding is influenced by the interplay between enthalpy and entropy contributions of the binding event. The impact of these binding free energy components, however, is not limited to the primary target only. Here, we investigate the relationship between binding thermodn. and selectivity profiles by combining publicly available data from broad off-target assay profiling and the corresponding thermodn. measurements. Our anal. indicates that compds. binding their primary targets with higher entropy contributions tend to hit more off-targets compared with those ligands that demonstrated enthalpy-driven binding.
37
Hopkins, A. L.; Keseru, G. M.; Leeson, P. D.; Rees, D. C.; Reynolds, C. H. Nat. Rev. Drug Discovery 2014, 13, 105– 121 DOI: 10.1038/nrd4163
Google Scholar
37
The role of ligand efficiency metrics in drug discovery
Hopkins, Andrew L.; Keserue, Gyoergy M.; Leeson, Paul D.; Rees, David C.; Reynolds, Charles H.
Nature Reviews Drug Discovery (2014), 13 (2), 105-121CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)
A review. The judicious application of ligand or binding efficiency metrics, which quantify the mol. properties required to obtain binding affinity for a drug target, is gaining traction in the selection and optimization of fragments, hits and leads. Retrospective anal. of recently marketed oral drugs shows that they frequently have highly optimized ligand efficiency values for their targets. Optimizing ligand efficiency metrics based on both mol. mass and lipophilicity, when set in the context of the specific target, has the potential to ameliorate the inflation of these properties that has been obsd. in current medicinal chem. practice, and to increase the quality of drug candidates.
38
Tummino, P. J.; Copeland, R. A. Biochemistry 2008, 47, 5481– 5492 DOI: 10.1021/bi8002023
Google Scholar
There is no corresponding record for this reference.
39
Perret, S.; Sabin, C.; Dumon, C.; Pokorná, M.; Gautier, C.; Galanina, O.; Ilia, S.; Bovin, N.; Nicaise, M.; Desmadril, M.; Gilboa-Garber, N.; Wimmerová, M.; Mitchell, E. P.; Imberty, A. Biochem. J. 2005, 389, 325– 332 DOI: 10.1042/BJ20050079
Google Scholar
There is no corresponding record for this reference.
40
Loris, R.; Tielker, D.; Jaeger, K.-E.; Wyns, L. J. Mol. Biol. 2003, 331, 861– 870 DOI: 10.1016/S0022-2836(03)00754-X
Google Scholar
There is no corresponding record for this reference.
41
Lagendijk, E. L.; Validov, S.; Lamers, G. E. M.; de Weert, S.; Bloemberg, G. V. FEMS Microbiol. Lett. 2010, 305, 81– 90 DOI: 10.1111/j.1574-6968.2010.01916.x
Google Scholar
There is no corresponding record for this reference.
42
Mayer, S.; Raulf, M.-K.; Lepenies, B. Histochem. Cell Biol. 2017, 147, 223– 237 DOI: 10.1007/s00418-016-1523-7
Google Scholar
42
C-type lectins: their network and roles in pathogen recognition and immunity
Mayer, Sabine; Raulf, Marie-Kristin; Lepenies, Bernd
Histochemistry and Cell Biology (2017), 147 (2), 223-237CODEN: HCBIFP; ISSN:0948-6143. (Springer)
C-type lectins (CTLs) represent the most complex family of animal/human lectins that comprises 17 different groups. During evolution, CTLs have developed by diversification to cover a broad range of glycan ligands. However, ligand binding by CTLs is not necessarily restricted to glycans as some CTLs also bind to proteins, lipids, inorg. mols., or ice crystals. CTLs share a common fold that harbors a Ca2+ for contact to the sugar and about 18 invariant residues in a phylogenetically conserved pattern. In vertebrates, CTLs have numerous functions, including serum glycoprotein homeostasis, pathogen sensing, and the initiation of immune responses. Myeloid CTLs in innate immunity are mainly expressed by antigen-presenting cells and play a prominent role in the recognition of a variety of pathogens such as fungi, bacteria, viruses, and parasites. However, myeloid CTLs such as the macrophage inducible CTL (Mincle) or Clec-9a may also bind to self-antigens and thus contribute to immune homeostasis. While some CTLs induce pro-inflammatory responses and thereby lead to activation of adaptive immune responses, other CTLs act as inhibitory receptors and dampen cellular functions. Since CTLs are key players in pathogen recognition and innate immunity, targeting CTLs may be a promising strategy for cell-specific delivery of drugs or vaccine antigens and to modulate immune responses.
43
Holla, A.; Skerra, A. Protein Eng., Des. Sel. 2011, 24, 659– 669 DOI: 10.1093/protein/gzr016
Google Scholar
43
Comparative analysis reveals selective recognition of glycans by the dendritic cell receptors DC-SIGN and Langerin
Holla, Andrea; Skerra, Arne
Protein Engineering, Design & Selection (2011), 24 (9), 659-669CODEN: PEDSBR; ISSN:1741-0126. (Oxford University Press)
DC-SIGN (dendritic cell-specific ICAM-3 grabbing non-integrin) and Langerin are homologous C-type lectins expressed as cell-surface receptors on different populations of dendritic cells (DCs). DC-SIGN interacts with glycan structures on HIV-1, facilitating virus survival, transmission and infection, whereas Langerin, which is characteristic of Langerhans cells (LCs), promotes HIV-1 uptake and degrdn. Here we describe a comprehensive comparison of the glycan specificities of both proteins by probing a synthetic carbohydrate microarray comprising 275 sugar compds. using the bacterially produced and fluorescence-labeled, monomeric carbohydrate-recognition domains (CRDs) of DC-SIGN and Langerin. In this side-by-side study DC-SIGN was found to preferentially bind internal mannose residues of high-mannose-type saccharides and the fucose-contg. blood-type antigens H, A, B, Lea, Leb Lex, Ley, sialyl-Lea as well as sulfatated derivs. of Lea and Lex. In contrast, Langerin appeared to recognize a different spectrum of compds., esp. those contg. terminal mannose, terminal N-acetylglucosamine and 6-sulfogalactose residues, but also the blood-type antigens H, A and B. Of the Lewis antigens, only Leb, Ley, sialyl-Lea and the sialyl-Lex deriv. with 6'-sulfatation at the galactose (sialyl-6SGal Lex) were weakly bound by Langerin. Notably, Ca2+-independent glycan-binding activity of Langerin could not be detected either by probing the glycan array or by isothermal titrn. calorimetry of the CRD with mannose and mannobiose. The precise knowledge of carbohydrate specificity of DC-SIGN and Langerin receptors resulting from our study may aid the future design of microbicides that specifically affect the DC-SIGN/HIV-1 interaction while not compromising the protective function of Langerin.
44
Wamhoff, E.-C.; Hanske, J.; Schnirch, L.; Aretz, J.; Grube, M.; Varón Silva, D.; Rademacher, C. ACS Chem. Biol. 2016, 11, 2407– 2413 DOI: 10.1021/acschembio.6b00561
Google Scholar
There is no corresponding record for this reference.
45
Zhang, Y.; Huo, M.; Zhou, J.; Xie, S. Comput. Methods Programs Biomed 2010, 99, 306– 314 DOI: 10.1016/j.cmpb.2010.01.007
Google Scholar
45
PKSolver: An add-in program for pharmacokinetic and pharmacodynamic data analysis in Microsoft Excel
Zhang Yong; Huo Meirong; Zhou Jianping; Xie Shaofei
Computer methods and programs in biomedicine (2010), 99 (3), 306-14 ISSN:.
This study presents PKSolver, a freely available menu-driven add-in program for Microsoft Excel written in Visual Basic for Applications (VBA), for solving basic problems in pharmacokinetic (PK) and pharmacodynamic (PD) data analysis. The program provides a range of modules for PK and PD analysis including noncompartmental analysis (NCA), compartmental analysis (CA), and pharmacodynamic modeling. Two special built-in modules, multiple absorption sites (MAS) and enterohepatic circulation (EHC), were developed for fitting the double-peak concentration-time profile based on the classical one-compartment model. In addition, twenty frequently used pharmacokinetic functions were encoded as a macro and can be directly accessed in an Excel spreadsheet. To evaluate the program, a detailed comparison of modeling PK data using PKSolver and professional PK/PD software package WinNonlin and Scientist was performed. The results showed that the parameters estimated with PKSolver were satisfactory. In conclusion, the PKSolver simplified the PK and PD data analysis process and its output could be generated in Microsoft Word in the form of an integrated report. The program provides pharmacokinetic researchers with a fast and easy-to-use tool for routine and basic PK and PD data analysis with a more user-friendly interface.
46
Allen, R. C.; Popat, R.; Diggle, S. P.; Brown, S. P. Nat. Rev. Microbiol. 2014, 12, 300– 308 DOI: 10.1038/nrmicro3232
Google Scholar
46
Targeting virulence: can we make evolution-proof drugs?
Allen, Richard C.; Popat, Roman; Diggle, Stephen P.; Brown, Sam P.
Nature Reviews Microbiology (2014), 12 (4), 300-308CODEN: NRMACK; ISSN:1740-1526. (Nature Publishing Group)
A review and discussion. Antivirulence drugs are a new type of therapeutic drug that target virulence factors, potentially revitalising the drug-development pipeline with new targets. As antivirulence drugs disarm the pathogen, rather than kill or halt pathogen growth, it has been hypothesized that they will generate much weaker selection for resistance than traditional antibiotics. However, recent studies have shown that mechanisms of resistance to antivirulence drugs exist, seemingly damaging the 'evolution-proof' claim. In this Opinion article, we highlight a crucial distinction between whether resistance can emerge and whether it will spread to a high frequency under drug selection. We argue that selection for resistance can be reduced, or even reversed, using appropriate combinations of target and treatment environment, opening a path towards the development of evolutionarily robust novel therapeutics.

Cited By

Click to copy section linkSection link copied!

Explore this article's citation statements on scite.ai

This article is cited by 109 publications.

Giulia Antonini, Mario Fares, Dirk Hauck, Patrycja Mała, Emilie Gillon, Laura Belvisi, Anna Bernardi, Alexander Titz, Annabelle Varrot, Sarah Mazzotta. Toward Dual-Target Glycomimetics against Two Bacterial Lectins to Fight Pseudomonas aeruginosa–Burkholderia cenocepacia Infections: A Biophysical Study. Journal of Medicinal Chemistry 2025, Article ASAP.
Eva Zahorska, Lisa Marie Denig, Stefan Lienenklaus, Sakonwan Kuhaudomlarp, Thomas Tschernig, Peter Lipp, Antje Munder, Emilie Gillon, Saverio Minervini, Varvara Verkhova, Anne Imberty, Stefanie Wagner, Alexander Titz. High-Affinity Lectin Ligands Enable the Detection of Pathogenic Pseudomonas aeruginosa Biofilms: Implications for Diagnostics and Therapy. JACS Au 2024, 4 (12) , 4715-4728. https://doi.org/10.1021/jacsau.4c00670
Daniel Kohnhäuser, Tim Seedorf, Katarina Cirnski, Dominik Heimann, Janetta Coetzee, Sylvie Sordello, Jana Richter, Moritz Stappert, Jean-Francois Sabuco, David Corbett, Eric Bacqué, Katharina Rox, Jennifer Herrmann, Aurelie Vassort, Rolf Müller, Andreas Kirschning, Mark Brönstrup. Optimization of the Central α-Amino Acid in Cystobactamids to the Broad-Spectrum, Resistance-Breaking Antibiotic CN-CC-861. Journal of Medicinal Chemistry 2024, 67 (19) , 17162-17190. https://doi.org/10.1021/acs.jmedchem.4c00927
Patrycja Mała, Eike Siebs, Joscha Meiers, Katharina Rox, Annabelle Varrot, Anne Imberty, Alexander Titz. Discovery of N-β-l-Fucosyl Amides as High-Affinity Ligands for the Pseudomonas aeruginosa Lectin LecB. Journal of Medicinal Chemistry 2022, 65 (20) , 14180-14200. https://doi.org/10.1021/acs.jmedchem.2c01373
Rafael Bermeo, Kanhaya Lal, Davide Ruggeri, Daniele Lanaro, Sarah Mazzotta, Francesca Vasile, Anne Imberty, Laura Belvisi, Annabelle Varrot, Anna Bernardi. Targeting a Multidrug-Resistant Pathogen: First Generation Antagonists of Burkholderia cenocepacia’s BC2L-C Lectin. ACS Chemical Biology 2022, 17 (10) , 2899-2910. https://doi.org/10.1021/acschembio.2c00532
Kartikey Singh, Suvarn S. Kulkarni. Small Carbohydrate Derivatives as Potent Antibiofilm Agents. Journal of Medicinal Chemistry 2022, 65 (13) , 8525-8549. https://doi.org/10.1021/acs.jmedchem.1c01039
Hashem Etayash, Morgan Alford, Noushin Akhoundsadegh, Matthew Drayton, Suzana K. Straus, Robert E. W. Hancock. Multifunctional Antibiotic–Host Defense Peptide Conjugate Kills Bacteria, Eradicates Biofilms, and Modulates the Innate Immune Response. Journal of Medicinal Chemistry 2021, 64 (22) , 16854-16863. https://doi.org/10.1021/acs.jmedchem.1c01712
Robert Wawrzinek, Eike-Christian Wamhoff, Jonathan Lefebre, Mareike Rentzsch, Gunnar Bachem, Gary Domeniconi, Jessica Schulze, Felix F. Fuchsberger, Hengxi Zhang, Carlos Modenutti, Lennart Schnirch, Marcelo A. Marti, Oliver Schwardt, Maria Bräutigam, Mónica Guberman, Dirk Hauck, Peter H. Seeberger, Oliver Seitz, Alexander Titz, Beat Ernst, Christoph Rademacher. A Remote Secondary Binding Pocket Promotes Heteromultivalent Targeting of DC-SIGN. Journal of the American Chemical Society 2021, 143 (45) , 18977-18988. https://doi.org/10.1021/jacs.1c07235
Joscha Meiers, Eva Zahorska, Teresa Röhrig, Dirk Hauck, Stefanie Wagner, Alexander Titz. Directing Drugs to Bugs: Antibiotic-Carbohydrate Conjugates Targeting Biofilm-Associated Lectins of Pseudomonas aeruginosa. Journal of Medicinal Chemistry 2020, 63 (20) , 11707-11724. https://doi.org/10.1021/acs.jmedchem.0c00856
Xiaoyan Ju, Jun Chen, Mengxue Zhou, Meng Zhu, Zhuang Li, Sijia Gao, Jinzhao Ou, Dandan Xu, Man Wu, Shidong Jiang, Yi Hu, Ye Tian, Zhongwei Niu. Combating Pseudomonas aeruginosa Biofilms by a Chitosan-PEG-Peptide Conjugate via Changes in Assembled Structure. ACS Applied Materials & Interfaces 2020, 12 (12) , 13731-13738. https://doi.org/10.1021/acsami.0c02034
Guoqing Tao, Tan Ji, Ning Wang, Guang Yang, Xiaolai Lei, Wei Zheng, Rongying Liu, Xuyang Xu, Ling Yang, Guang-Qiang Yin, Xiaojuan Liao, Xiaopeng Li, Hong-ming Ding, Xiaoming Ding, Jinfu Xu, Hai-Bo Yang, Guosong Chen. Self-Assembled Saccharide-Functionalized Amphiphilic Metallacycles as Biofilms Inhibitor via “Sweet Talking”. ACS Macro Letters 2020, 9 (1) , 61-69. https://doi.org/10.1021/acsmacrolett.9b00914
Yu Zhao, Cong Yu, Yunjian Yu, Xiaosong Wei, Xiaozhuang Duan, Xijuan Dai, Xinge Zhang. Bioinspired Heteromultivalent Ligand-Decorated Nanotherapeutic for Enhanced Photothermal and Photodynamic Therapy of Antibiotic-Resistant Bacterial Pneumonia. ACS Applied Materials & Interfaces 2019, 11 (43) , 39648-39661. https://doi.org/10.1021/acsami.9b15118
Jonathan Cramer, Christoph P. Sager, Beat Ernst. Hydroxyl Groups in Synthetic and Natural-Product-Derived Therapeutics: A Perspective on a Common Functional Group. Journal of Medicinal Chemistry 2019, 62 (20) , 8915-8930. https://doi.org/10.1021/acs.jmedchem.9b00179
Roman Sommer, Katharina Rox, Stefanie Wagner, Dirk Hauck, Sarah S. Henrikus, Shelby Newsad, Tatjana Arnold, Thomas Ryckmans, Mark Brönstrup, Anne Imberty, Annabelle Varrot, Rolf W. Hartmann, Alexander Titz. Anti-biofilm Agents against Pseudomonas aeruginosa: A Structure–Activity Relationship Study of C-Glycosidic LecB Inhibitors. Journal of Medicinal Chemistry 2019, 62 (20) , 9201-9216. https://doi.org/10.1021/acs.jmedchem.9b01120
Marwa Taouai, Khouloud Chakroun, Roman Sommer, Gaelle Michaud, David Giacalone, Mohamed Amine Ben Maaouia, Aurélie Vallin-Butruille, David Mathiron, Rym Abidi, Tamis Darbre, Peter J. Cragg, Catherine Mullié, Jean-Louis Reymond, George A. O’Toole, Mohammed Benazza. Glycocluster Tetrahydroxamic Acids Exhibiting Unprecedented Inhibition of Pseudomonas aeruginosa Biofilms. Journal of Medicinal Chemistry 2019, 62 (17) , 7722-7738. https://doi.org/10.1021/acs.jmedchem.9b00481
Eike-Christian Wamhoff, Jessica Schulze, Lydia Bellmann, Mareike Rentzsch, Gunnar Bachem, Felix F. Fuchsberger, Juliane Rademacher, Martin Hermann, Barbara Del Frari, Rob van Dalen, David Hartmann, Nina M. van Sorge, Oliver Seitz, Patrizia Stoitzner, Christoph Rademacher. A Specific, Glycomimetic Langerin Ligand for Human Langerhans Cell Targeting. ACS Central Science 2019, 5 (5) , 808-820. https://doi.org/10.1021/acscentsci.9b00093
Stéphane Baeriswyl, Bee-Ha Gan, Thissa N. Siriwardena, Ricardo Visini, Maurane Robadey, Sacha Javor, Achim Stocker, Tamis Darbre, Jean-Louis Reymond. X-ray Crystal Structures of Short Antimicrobial Peptides as Pseudomonas aeruginosa Lectin B Complexes. ACS Chemical Biology 2019, 14 (4) , 758-766. https://doi.org/10.1021/acschembio.9b00047
François Portier, Anne Imberty, Sami Halila. Expeditious Synthesis of C-Glycosyl Barbiturate Ligands of Bacterial Lectins: From Monomer Design to Glycoclusters and Glycopolymers. Bioconjugate Chemistry 2019, 30 (3) , 647-656. https://doi.org/10.1021/acs.bioconjchem.8b00847
Tamir Dingjan, Émilie Gillon, Anne Imberty, Serge Pérez, Alexander Titz, Paul A. Ramsland, Elizabeth Yuriev. Virtual Screening Against Carbohydrate-Binding Proteins: Evaluation and Application to Bacterial Burkholderia ambifaria Lectin. Journal of Chemical Information and Modeling 2018, 58 (9) , 1976-1989. https://doi.org/10.1021/acs.jcim.8b00185
Yincheng Chang, Zehuan Huang, Yang Jiao, Jiang-Fei Xu, Xi Zhang. pH-Induced Charge-Reversal Amphiphile with Cancer Cell-Selective Membrane-Disrupting Activity. ACS Applied Materials & Interfaces 2018, 10 (25) , 21191-21197. https://doi.org/10.1021/acsami.8b06660
Elena Di Marzo, Luigi Lay, Giuseppe D’Orazio. Synthesis of N,N-Dimethylaminopropyl Derivative of A Blood Sugar Antigen. Molbank 2025, 2025 (2) , M1985. https://doi.org/10.3390/M1985
Kathrin Siebold, Elena Chikunova, Nils Lorz, Christina Jordan, Alvar D. Gossert, Ryan Gilmour. Fluor‐Fucosylierung ermöglicht die Untersuchung der Le a ‐LecB‐Wechselwirkung durch BioNMR‐Spektroskopie. Angewandte Chemie 2025, 137 (11) https://doi.org/10.1002/ange.202423782
Kathrin Siebold, Elena Chikunova, Nils Lorz, Christina Jordan, Alvar D. Gossert, Ryan Gilmour. Fluoro‐Fucosylation Enables the Interrogation of the Le a ‐LecB Interaction by BioNMR Spectroscopy. Angewandte Chemie International Edition 2025, 64 (11) https://doi.org/10.1002/anie.202423782
Wenjing Ma, Junyuan Luo, Huan Liu, Qi Du, Tianhui Hao, Yinyu Jiang, Zhengwei Huang, Lefu Lan, Zhonghua Li, Tiehai Li. Chemoenzymatic Synthesis of Highly O ‐Glycosylated MUC7 Glycopeptides for Probing Inhibitory Activity against Pseudomonas aeruginosa Biofilm Formation. Angewandte Chemie 2025, 33 https://doi.org/10.1002/ange.202424312
Wenjing Ma, Junyuan Luo, Huan Liu, Qi Du, Tianhui Hao, Yinyu Jiang, Zhengwei Huang, Lefu Lan, Zhonghua Li, Tiehai Li. Chemoenzymatic Synthesis of Highly O ‐Glycosylated MUC7 Glycopeptides for Probing Inhibitory Activity against Pseudomonas aeruginosa Biofilm Formation. Angewandte Chemie International Edition 2025, 33 https://doi.org/10.1002/anie.202424312
Giuseppe D’Orazio, Barbara La Ferla. Synthesis of a Hydrogen Isotope-Labeled SGLT1 C-Glucoside Ligand for Distribution and Metabolic Fate Studies. Molbank 2025, 2025 (1) , M1982. https://doi.org/10.3390/M1982
José L. de Paz, Pedro M. Nieto. Fluorescence polarization assays to study carbohydrate–protein interactions. Organic & Biomolecular Chemistry 2025, 23 (9) , 2041-2058. https://doi.org/10.1039/D4OB02021A
Ghamdan Beshr, Asfandyar Sikandar, Julia Gläser, Mario Fares, Roman Sommer, Stefanie Wagner, Jesko Köhnke, Alexander Titz. A fucose-binding superlectin from Enterobacter cloacae with high Lewis and ABO blood group antigen specificity. Journal of Biological Chemistry 2025, 301 (2) , 108151. https://doi.org/10.1016/j.jbc.2024.108151
Marie Hanot, Elodie Lohou, Pascal Sonnet. Anti-Biofilm Agents to Overcome Pseudomonas aeruginosa Antibiotic Resistance. Pharmaceuticals 2025, 18 (1) , 92. https://doi.org/10.3390/ph18010092
Helma David, Sahana Vasudevan, Adline Princy Solomon. Mitigating candidiasis with acarbose by targeting Candida albicans α-glucosidase: in-silico, in-vitro and transcriptomic approaches. Scientific Reports 2024, 14 (1) https://doi.org/10.1038/s41598-024-62684-x
Paul V. Murphy, Ashis Dhara, Liam S. Fitzgerald, Eoin Hever, Saidulu Konda, Kishan Mandal. Small lectin ligands as a basis for applications in glycoscience and glycomedicine. Chemical Society Reviews 2024, 53 (19) , 9428-9445. https://doi.org/10.1039/D4CS00642A
Chenxiao Wan, Xiaoyan Ju, Dandan Xu, Jinzhao Ou, Meng Zhu, Guojun Lu, Kejia Li, Wei Jiang, Chunyan Li, Xiaohua Hu, Ye Tian, Zhongwei Niu. Escherichia coli exopolysaccharides disrupt Pseudomonas aeruginosa biofilm and increase its antibiotic susceptibility. Acta Biomaterialia 2024, 185 , 215-225. https://doi.org/10.1016/j.actbio.2024.07.028
Christian Vogel, Katharina Rox, Florian Wagenlehner, Alexander Titz. Glycomimetics as Candidates for Treatment and Prevention of Catheter-associated Biofilms Formed by Pseudomonas aeruginosa. European Urology Focus 2024, 10 (5) , 720-721. https://doi.org/10.1016/j.euf.2024.08.011
Lorina Badger-Emeka, Promise Emeka, Krishnaraj Thirugnanasambantham, Abdulaziz S. Alatawi. The Role of Pseudomonas aeruginosa in the Pathogenesis of Corneal Ulcer, Its Associated Virulence Factors, and Suggested Novel Treatment Approaches. Pharmaceutics 2024, 16 (8) , 1074. https://doi.org/10.3390/pharmaceutics16081074
Qiang Zhang, Laurent Soulère, Yves Queneau. Amide bioisosteric replacement in the design and synthesis of quorum sensing modulators. European Journal of Medicinal Chemistry 2024, 273 , 116525. https://doi.org/10.1016/j.ejmech.2024.116525
Truong Thanh Tung, Nguyen Quoc Thang, Nguyen Cao Huy, Pham Bao Phuong, Dinh Ngoc Minh, Nguyen Hai Nam, John Nielsen. Identification of novel phenylalanine derivatives bearing a hydroxamic acid moiety as potent quorum sensing inhibitors. RSC Medicinal Chemistry 2024, 15 (4) , 1320-1328. https://doi.org/10.1039/D3MD00670K
Deborah L. Chance, Wei Wang, James K. Waters, Thomas P. Mawhinney. Insights on Pseudomonas aeruginosa Carbohydrate Binding from Profiles of Cystic Fibrosis Isolates Using Multivalent Fluorescent Glycopolymers Bearing Pendant Monosaccharides. Microorganisms 2024, 12 (4) , 801. https://doi.org/10.3390/microorganisms12040801
Dong Yu, Zhaoyu Lu, Fengsong Nie, Yang Chong. Integrins regulation of wound healing processes: insights for chronic skin wound therapeutics. Frontiers in Cellular and Infection Microbiology 2024, 14 https://doi.org/10.3389/fcimb.2024.1324441
Êmily Évelyn Bandeira Batista, Marcelo Antônio de Souza Silva, José Lucas Medeiros Torres, Waldo Silva Mariz, Vanessa Beatriz Jales Rego, André Felipe Dutra Leitão, Abrahão Alves de Oliveira Filho, Veneziano Guedes de Sousa Rêgo. AVALIAÇÃO DA ATIVIDADE ANTIBACTERIANA E ANTIADERENTE DO ÓLEO ESSENCIAL DE Lavandula hybrida grosso CONTRA Pseudomonas aeruginosa. Revista Multidisciplinar do Nordeste Mineiro 2024, 1 (1) https://doi.org/10.61164/rmnm.v1i1.2084
Dong Yu, Zhaoyu Lu, Yang Chong. Integrins as a bridge between bacteria and cells: key targets for therapeutic wound healing. Burns & Trauma 2024, 12 https://doi.org/10.1093/burnst/tkae022
Ting Du, Jiangli Cao, Zhannuo Zhang, Zehui Xiao, Jingbo Jiao, Zhiyong Song, Xinjun Du, Shuo Wang. Thermo-responsive cascade antimicrobial platform for precise biofilm removal and enhanced wound healing. Burns & Trauma 2024, 12 https://doi.org/10.1093/burnst/tkae038
Yu Fan, Ahmed El Rhaz, Stéphane Maisonneuve, Emilie Gillon, Maha Fatthalla, Franck Le Bideau, Guillaume Laurent, Samir Messaoudi, Anne Imberty, Juan Xie. Photoswitchable glycoligands targeting Pseudomonas aeruginosa LecA. Beilstein Journal of Organic Chemistry 2024, 20 , 1486-1496. https://doi.org/10.3762/bjoc.20.132
Jonathan Cramer1,2, Lijuan Pang3, Beat Ernst1. Antiadhesive Carbohydrates and Glycomimetics. 2023, 131-160. https://doi.org/10.1002/9783527831326.ch5
Qianqian Guo, Yijie Cheng, Zheng Fan, Weihui Wu, Zhongming Wu, Xinge Zhang. Zwitterion‐conjugated Topological Glycomimics for Dual‐Blocking Effects to Eradicate Biofilm Infection. Advanced Therapeutics 2023, 6 (12) https://doi.org/10.1002/adtp.202300217
Xiaoxia Kang, Xiaoxiao Yang, Yue He, Conglin Guo, Yuechen Li, Haiwei Ji, Yuling Qin, Li Wu. Strategies and materials for the prevention and treatment of biofilms. Materials Today Bio 2023, 23 , 100827. https://doi.org/10.1016/j.mtbio.2023.100827
Rajendra Prasad Janapatla, Anna Dudek, Chyi-Liang Chen, Chih-Hsien Chuang, Kun-Yi Chien, Ye Feng, Yuan-Ming Yeh, Yi-Hsin Wang, Hsin-Ju Chang, Yuan-Chuan Lee, Cheng-Hsun Chiu. Marine prebiotics mediate decolonization of Pseudomonas aeruginosa from gut by inhibiting secreted virulence factor interactions with mucins and enriching Bacteroides population. Journal of Biomedical Science 2023, 30 (1) https://doi.org/10.1186/s12929-023-00902-w
Jamuna Bai Aswathanarayan, Ravishankar Rai Vittal. Small molecules as next generation biofilm inhibitors and anti-infective agents. Physical Sciences Reviews 2023, 8 (11) , 4361-4373. https://doi.org/10.1515/psr-2021-0175
Aghiad Bali, Mohamed A. M. Kamal, Glorjen Mulla, Brigitta Loretz, Claus‐Michael Lehr. Functional Materials to Overcome Bacterial Barriers and Models to Advance Their Development. Advanced Functional Materials 2023, 33 (45) https://doi.org/10.1002/adfm.202304370
Nikhil Sathe, Peter Beech, Larry Croft, Cenk Suphioglu, Arnab Kapat, Eugene Athan. Pseudomonas aeruginosa: Infections and novel approaches to treatment “Knowing the enemy” the threat of Pseudomonas aeruginosa and exploring novel approaches to treatment. Infectious Medicine 2023, 2 (3) , 178-194. https://doi.org/10.1016/j.imj.2023.05.003
Sandra Behren, Jin Yu, Christian Pett, Manuel Schorlemer, Viktoria Heine, Thomas Fischöder, Lothar Elling, Ulrika Westerlind. Fucose Binding Motifs on Mucin Core Glycopeptides Impact Bacterial Lectin Recognition**. Angewandte Chemie 2023, 135 (32) https://doi.org/10.1002/ange.202302437
Sandra Behren, Jin Yu, Christian Pett, Manuel Schorlemer, Viktoria Heine, Thomas Fischöder, Lothar Elling, Ulrika Westerlind. Fucose Binding Motifs on Mucin Core Glycopeptides Impact Bacterial Lectin Recognition**. Angewandte Chemie International Edition 2023, 62 (32) https://doi.org/10.1002/anie.202302437
Steffen Leusmann, Petra Ménová, Elena Shanin, Alexander Titz, Christoph Rademacher. Glycomimetics for the inhibition and modulation of lectins. Chemical Society Reviews 2023, 52 (11) , 3663-3740. https://doi.org/10.1039/D2CS00954D
Gareth LuTheryn, Elaine M. L. Ho, Victor Choi, Dario Carugo. Cationic Microbubbles for Non-Selective Binding of Cavitation Nuclei to Bacterial Biofilms. Pharmaceutics 2023, 15 (5) , 1495. https://doi.org/10.3390/pharmaceutics15051495
Janina Sponsel, Yubing Guo, Lutfir Hamzam, Alice C Lavanant, Annia Pérez‐Riverón, Emma Partiot, Quentin Muller, Julien Rottura, Raphael Gaudin, Dirk Hauck, Alexander Titz, Vincent Flacher, Winfried Römer, Christopher G Mueller. Pseudomonas aeruginosa LecB suppresses immune responses by inhibiting transendothelial migration. EMBO reports 2023, 24 (4) https://doi.org/10.15252/embr.202255971
Eva Zahorska, Francesca Rosato, Kai Stober, Sakonwan Kuhaudomlarp, Joscha Meiers, Dirk Hauck, Dorina Reith, Emilie Gillon, Katharina Rox, Anne Imberty, Winfried Römer, Alexander Titz. Neutralisation der Auswirkungen des Virulenzfaktors LecA aus Pseudomonas aeruginosa auf Humanzellen durch neue glykomimetische Inhibitoren. Angewandte Chemie 2023, 135 (7) https://doi.org/10.1002/ange.202215535
Eva Zahorska, Francesca Rosato, Kai Stober, Sakonwan Kuhaudomlarp, Joscha Meiers, Dirk Hauck, Dorina Reith, Emilie Gillon, Katharina Rox, Anne Imberty, Winfried Römer, Alexander Titz. Neutralizing the Impact of the Virulence Factor LecA from Pseudomonas aeruginosa on Human Cells with New Glycomimetic Inhibitors. Angewandte Chemie International Edition 2023, 62 (7) https://doi.org/10.1002/anie.202215535
Ana Marta de Matos. Recent Advances in the Development and Synthesis of Carbohydrate‐Based Molecules with Promising Antibacterial Activity. European Journal of Organic Chemistry 2023, 26 (4) https://doi.org/10.1002/ejoc.202200919
David Elder. Challenges in the development of novel antibiotics. 2023, 65-85. https://doi.org/10.1016/B978-0-323-95388-7.00005-X
Lukas Gajdos, Matthew P. Blakeley, Michael Haertlein, V. Trevor Forsyth, Juliette M. Devos, Anne Imberty. Neutron crystallography reveals mechanisms used by Pseudomonas aeruginosa for host-cell binding. Nature Communications 2022, 13 (1) https://doi.org/10.1038/s41467-021-27871-8
Oliver Riester, Pia Burkhardtsmaier, Yuna Gurung, Stefan Laufer, Hans-Peter Deigner, Magnus S. Schmidt. Synergy of R-(–)carvone and cyclohexenone-based carbasugar precursors with antibiotics to enhance antibiotic potency and inhibit biofilm formation. Scientific Reports 2022, 12 (1) https://doi.org/10.1038/s41598-022-22807-8
Elena Shanina, Sakonwan Kuhaudomlarp, Eike Siebs, Felix F. Fuchsberger, Maxime Denis, Priscila da Silva Figueiredo Celestino Gomes, Mads H. Clausen, Peter H. Seeberger, Didier Rognan, Alexander Titz, Anne Imberty, Christoph Rademacher. Targeting undruggable carbohydrate recognition sites through focused fragment library design. Communications Chemistry 2022, 5 (1) https://doi.org/10.1038/s42004-022-00679-3
Magdolna Csávás, László Kalmár, Petronella Szőke, László Bence Farkas, Bálint Bécsi, Zoltán Kónya, János Kerékgyártó, Anikó Borbás, Ferenc Erdődi, Katalin E. Kövér. A Fucosylated Lactose-Presenting Tetravalent Glycocluster Acting as a Mutual Ligand of Pseudomonas aeruginosa Lectins A (PA-IL) and B (PA-IIL)—Synthesis and Interaction Studies. International Journal of Molecular Sciences 2022, 23 (24) , 16194. https://doi.org/10.3390/ijms232416194
Jack A. Doolan, George T. Williams, Kira L. F. Hilton, Rajas Chaudhari, John S. Fossey, Benjamin T. Goult, Jennifer R. Hiscock. Advancements in antimicrobial nanoscale materials and self-assembling systems. Chemical Society Reviews 2022, 51 (20) , 8696-8755. https://doi.org/10.1039/D1CS00915J
Andreja Jakas, Ramya Ayyalasomayajula, Mare Cudic, Ivanka Jerić. Multicomponent reaction derived small di- and tri-carbohydrate-based glycomimetics as tools for probing lectin specificity. Glycoconjugate Journal 2022, 39 (5) , 587-597. https://doi.org/10.1007/s10719-022-10079-3
Marwa Fray, David Mathiron, Serge Pilard, David Lesur, Rym Abidi, Thouraya Barhoumi‐Slimi, Peter J. Cragg, Mohammed Benazza. Heteroglycoclusters through Unprecedented Orthogonal Chemistry Based on N ‐Alkylation of N ‐Acylhydrazone. European Journal of Organic Chemistry 2022, 2022 (33) https://doi.org/10.1002/ejoc.202101537
Delia Boffoli, Federica Bellato, Greta Avancini, Pratik Gurnani, Gokhan Yilmaz, Manuel Romero, Shaun Robertson, Francesca Moret, Federica Sandrelli, Paolo Caliceti, Stefano Salmaso, Miguel Cámara, Giuseppe Mantovani, Francesca Mastrotto. Tobramycin-loaded complexes to prevent and disrupt Pseudomonas aeruginosa biofilms. Drug Delivery and Translational Research 2022, 12 (8) , 1788-1810. https://doi.org/10.1007/s13346-021-01085-3
Yinglu Wang, Lile Pan, Li Li, Ruipin Cao, Qian Zheng, Zuxian Xu, Chang-Jer Wu, Hu Zhu. Glycosylation increases the anti-QS as well as anti-biofilm and anti-adhesion ability of the cyclo (L-Trp-L-Ser) against Pseudomonas aeruginosa. European Journal of Medicinal Chemistry 2022, 238 , 114457. https://doi.org/10.1016/j.ejmech.2022.114457
Karolina Wojtczak, Joseph P. Byrne. Structural Considerations for Building Synthetic Glycoconjugates as Inhibitors for Pseudomonas aeruginosa Lectins. ChemMedChem 2022, 17 (12) https://doi.org/10.1002/cmdc.202200081
Gheorghe Roman. Thiophene‐containing compounds with antimicrobial activity. Archiv der Pharmazie 2022, 355 (6) https://doi.org/10.1002/ardp.202100462
Yiwen Zhu, Shijing Wu, Yujie Sun, Xuan Zou, Liang Zheng, Shun Duan, Julin Wang, Bingran Yu, Ruifang Sui, Fu‐Jian Xu. Bacteria‐Targeting Photodynamic Nanoassemblies for Efficient Treatment of Multidrug‐Resistant Biofilm Infected Keratitis. Advanced Functional Materials 2022, 32 (14) https://doi.org/10.1002/adfm.202111066
Eike Siebs, Elena Shanina, Sakonwan Kuhaudomlarp, Priscila da Silva Figueiredo Celestino Gomes, Cloé Fortin, Peter H. Seeberger, Didier Rognan, Christoph Rademacher, Anne Imberty, Alexander Titz. Targeting the Central Pocket of the Pseudomonas aeruginosa Lectin LecA. ChemBioChem 2022, 23 (3) https://doi.org/10.1002/cbic.202100563
Mostafa Dadashi Firouzjaei, Mehdi Pejman, Mohammad Sharifian Gh, Sadegh Aghapour Aktij, Ehsan Zolghadr, Ahmad Rahimpour, Mohtada Sadrzadeh, Ahmad Arabi Shamsabadi, Alberto Tiraferri, Mark Elliott. Functionalized polyamide membranes yield suppression of biofilm and planktonic bacteria while retaining flux and selectivity. Separation and Purification Technology 2022, 282 , 119981. https://doi.org/10.1016/j.seppur.2021.119981
Olga Metelkina, Benedikt Huck, Jonathan S. O'Connor, Marcus Koch, Andreas Manz, Claus-Michael Lehr, Alexander Titz. Targeting extracellular lectins of Pseudomonas aeruginosa with glycomimetic liposomes. Journal of Materials Chemistry B 2022, 10 (4) , 537-548. https://doi.org/10.1039/D1TB02086B
Yi Zhao, Long Chen, Yanan Wang, Xinyu Song, Keyang Li, Xuefeng Yan, Liangmin Yu, Zhiyu He. Nanomaterial-based strategies in antimicrobial applications: Progress and perspectives. Nano Research 2021, 14 (12) , 4417-4441. https://doi.org/10.1007/s12274-021-3417-4
Truong Thanh Tung, Huy Luong Xuan. “Left-hand strategy” for the design, synthesis and discovery of novel triazole–mercaptobenzothiazole hybrid compounds as potent quorum sensing inhibitors and anti-biofilm formation of Pseudomonas aeruginosa. New Journal of Chemistry 2021, 45 (46) , 21631-21637. https://doi.org/10.1039/D1NJ04436B
Ana M. Gomez, J. Cristobal Lopez. Bringing Color to Sugars: The Chemical Assembly of Carbohydrates to BODIPY Dyes. The Chemical Record 2021, 21 (11) , 3112-3130. https://doi.org/10.1002/tcr.202100190
Ciamak Ghazaei. Anti-virulence Therapy Against Bacterial Infections: Mechanisms of Action and Challenges. Journal of Kermanshah University of Medical Sciences 2021, 25 (3) https://doi.org/10.5812/jkums.111808
Marcus Miethke, Marco Pieroni, Tilmann Weber, Mark Brönstrup, Peter Hammann, Ludovic Halby, Paola B. Arimondo, Philippe Glaser, Bertrand Aigle, Helge B. Bode, Rui Moreira, Yanyan Li, Andriy Luzhetskyy, Marnix H. Medema, Jean-Luc Pernodet, Marc Stadler, José Rubén Tormo, Olga Genilloud, Andrew W. Truman, Kira J. Weissman, Eriko Takano, Stefano Sabatini, Evi Stegmann, Heike Brötz-Oesterhelt, Wolfgang Wohlleben, Myriam Seemann, Martin Empting, Anna K. H. Hirsch, Brigitta Loretz, Claus-Michael Lehr, Alexander Titz, Jennifer Herrmann, Timo Jaeger, Silke Alt, Thomas Hesterkamp, Mathias Winterhalter, Andrea Schiefer, Kenneth Pfarr, Achim Hoerauf, Heather Graz, Michael Graz, Mika Lindvall, Savithri Ramurthy, Anders Karlén, Maarten van Dongen, Hrvoje Petkovic, Andreas Keller, Frédéric Peyrane, Stefano Donadio, Laurent Fraisse, Laura J. V. Piddock, Ian H. Gilbert, Heinz E. Moser, Rolf Müller. Towards the sustainable discovery and development of new antibiotics. Nature Reviews Chemistry 2021, 5 (10) , 726-749. https://doi.org/10.1038/s41570-021-00313-1
Tung Truong Thanh, Huy Luong Xuan, Thang Nguyen Quoc. Benzo[ d ]thiazole-2-thiol bearing 2-oxo-2-substituted-phenylethan-1-yl as potent selective lasB quorum sensing inhibitors of Gram-negative bacteria. RSC Advances 2021, 11 (46) , 28797-28808. https://doi.org/10.1039/D1RA03616E
Michael A. Trebino, Rahul D. Shingare, John B. MacMillan, Fitnat H. Yildiz. Strategies and Approaches for Discovery of Small Molecule Disruptors of Biofilm Physiology. Molecules 2021, 26 (15) , 4582. https://doi.org/10.3390/molecules26154582
Vishnu C. Damalanka, Amarendar Reddy Maddirala, James W. Janetka. Novel approaches to glycomimetic design: development of small molecular weight lectin antagonists. Expert Opinion on Drug Discovery 2021, 16 (5) , 513-536. https://doi.org/10.1080/17460441.2021.1857721
Roland E. Dolle, Laurel K. McGrane, James W. Janetka, Jared T. Hammill, Rodney Kiplin Guy, Anjan Debnath, Peter M. Cheuka, Claire Le Manach, Kelly Chibale, Ake Elhammer, Martin Handfield, Michael J. Dawson, Stefano Donadio, Jae H. Park, Alan Joslyn. Anti‐Infective Case Histories. 2021, 1-95. https://doi.org/10.1002/0471266949.bmc288
Sakonwan Kuhaudomlarp, Eike Siebs, Elena Shanina, Jérémie Topin, Ines Joachim, Priscila da Silva Figueiredo Celestino Gomes, Annabelle Varrot, Didier Rognan, Christoph Rademacher, Anne Imberty, Alexander Titz. Non‐Carbohydrate Glycomimetics as Inhibitors of Calcium(II)‐Binding Lectins. Angewandte Chemie 2021, 133 (15) , 8185-8195. https://doi.org/10.1002/ange.202013217
Sakonwan Kuhaudomlarp, Eike Siebs, Elena Shanina, Jérémie Topin, Ines Joachim, Priscila da Silva Figueiredo Celestino Gomes, Annabelle Varrot, Didier Rognan, Christoph Rademacher, Anne Imberty, Alexander Titz. Non‐Carbohydrate Glycomimetics as Inhibitors of Calcium(II)‐Binding Lectins. Angewandte Chemie International Edition 2021, 60 (15) , 8104-8114. https://doi.org/10.1002/anie.202013217
Nives Hribernik, Alice Tamburrini, Ermelinda Falletta, Anna Bernardi. One pot synthesis of thio -glycosides via aziridine opening reactions. Organic & Biomolecular Chemistry 2021, 19 (1) , 233-247. https://doi.org/10.1039/D0OB01956A
Jocelyn A. Hammond, Emma A. Gordon, Kayla M. Socarras, Joshua Chang Mell, Garth D. Ehrlich. Beyond the pan-genome: current perspectives on the functional and practical outcomes of the distributed genome hypothesis. Biochemical Society Transactions 2020, 48 (6) , 2437-2455. https://doi.org/10.1042/BST20190713
Isabelle Bertin-Jung, Anne Robert, Nick Ramalanjaona, Sandrine Gulberti, Catherine Bui, Jean-Baptiste Vincourt, Mohamed Ouzzine, Jean-Claude Jacquinet, Chrystel Lopin-Bon, Sylvie Fournel-Gigleux. A versatile strategy to synthesize N -methyl-anthranilic acid-labelled glycoprobes for fluorescence-based screening assays. Chemical Communications 2020, 56 (73) , 10746-10749. https://doi.org/10.1039/D0CC03882B
Carmen S. Padilla, Mona B. Damaj, Zhong-Nan Yang, Joe Molina, Brian R. Berquist, Earl L. White, Nora Solís-Gracia, Jorge Da Silva, Kranthi K. Mandadi. High-Level Production of Recombinant Snowdrop Lectin in Sugarcane and Energy Cane. Frontiers in Bioengineering and Biotechnology 2020, 8 https://doi.org/10.3389/fbioe.2020.00977
Eva Zahorska, Sakonwan Kuhaudomlarp, Saverio Minervini, Sultaan Yousaf, Martin Lepsik, Thorsten Kinsinger, Anna K. H. Hirsch, Anne Imberty, Alexander Titz. A rapid synthesis of low-nanomolar divalent LecA inhibitors in four linear steps from d -galactose pentaacetate. Chemical Communications 2020, 56 (62) , 8822-8825. https://doi.org/10.1039/D0CC03490H
Roland Thuenauer, Alessia Landi, Anne Trefzer, Silke Altmann, Sarah Wehrum, Thorsten Eierhoff, Britta Diedrich, Jörn Dengjel, Alexander Nyström, Anne Imberty, Winfried Römer, , . The Pseudomonas aeruginosa Lectin LecB Causes Integrin Internalization and Inhibits Epithelial Wound Healing. mBio 2020, 11 (2) https://doi.org/10.1128/mBio.03260-19
Joscha Meiers, Eike Siebs, Eva Zahorska, Alexander Titz. Lectin antagonists in infection, immunity, and inflammation. Current Opinion in Chemical Biology 2019, 53 , 51-67. https://doi.org/10.1016/j.cbpa.2019.07.005
Daniel Passos da Silva, Michael L. Matwichuk, Delaney O. Townsend, Courtney Reichhardt, Doriano Lamba, Daniel J. Wozniak, Matthew R. Parsek. The Pseudomonas aeruginosa lectin LecB binds to the exopolysaccharide Psl and stabilizes the biofilm matrix. Nature Communications 2019, 10 (1) https://doi.org/10.1038/s41467-019-10201-4
Alessia Landi, Muriel Mari, Svenja Kleiser, Tobias Wolf, Christine Gretzmeier, Isabel Wilhelm, Dimitra Kiritsi, Roland Thünauer, Roger Geiger, Alexander Nyström, Fulvio Reggiori, Julie Claudinon, Winfried Römer. Pseudomonas aeruginosa lectin LecB impairs keratinocyte fitness by abrogating growth factor signalling. Life Science Alliance 2019, 2 (6) , e201900422. https://doi.org/10.26508/lsa.201900422
Pablo Valverde, Ana Ardá, Niels-Christian Reichardt, Jesús Jiménez-Barbero, Ana Gimeno. Glycans in drug discovery. MedChemComm 2019, 10 (10) , 1678-1691. https://doi.org/10.1039/C9MD00292H
Stéphane Baeriswyl, Sacha Javor, Achim Stocker, Tamis Darbre, Jean‐Louis Reymond. X‐Ray Crystal Structure of a Second‐Generation Peptide Dendrimer in Complex with Pseudomonas aeruginosa Lectin LecB. Helvetica Chimica Acta 2019, 102 (9) https://doi.org/10.1002/hlca.201900178
Martin Lepsik, Roman Sommer, Sakonwan Kuhaudomlarp, Mickaël Lelimousin, Emanuele Paci, Annabelle Varrot, Alexander Titz, Anne Imberty. Induction of rare conformation of oligosaccharide by binding to calcium-dependent bacterial lectin: X-ray crystallography and modelling study. European Journal of Medicinal Chemistry 2019, 177 , 212-220. https://doi.org/10.1016/j.ejmech.2019.05.049
Zhengwei Liu, Faming Wang, Jinsong Ren, Xiaogang Qu. A series of MOF/Ce-based nanozymes with dual enzyme-like activity disrupting biofilms and hindering recolonization of bacteria. Biomaterials 2019, 208 , 21-31. https://doi.org/10.1016/j.biomaterials.2019.04.007
Son Thai Le, Lenka Malinovska, Michaela Vašková, Erika Mező, Viktor Kelemen, Anikó Borbás, Petr Hodek, Michaela Wimmerová, Magdolna Csávás. Investigation of the Binding Affinity of a Broad Array of l-Fucosides with Six Fucose-Specific Lectins of Bacterial and Fungal Origin. Molecules 2019, 24 (12) , 2262. https://doi.org/10.3390/molecules24122262
Rachel Hevey. Strategies for the Development of Glycomimetic Drug Candidates. Pharmaceuticals 2019, 12 (2) , 55. https://doi.org/10.3390/ph12020055
Jo Sing Julia Tang, Sophia Rosencrantz, Lucas Tepper, Sany Chea, Stefanie Klöpzig, Anne Krüger-Genge, Joachim Storsberg, Ruben R. Rosencrantz. Functional Glyco-Nanogels for Multivalent Interaction with Lectins. Molecules 2019, 24 (10) , 1865. https://doi.org/10.3390/molecules24101865

Load all citations

Download PDF

Get e-Alerts

Journal of the American Chemical Society

Cite this: J. Am. Chem. Soc. 2018, 140, 7, 2537–2545

Click to copy citationCitation copied!

https://doi.org/10.1021/jacs.7b11133

Published December 22, 2017

Request reuse permissions

Article Views

Altmetric

Citations

109

Learn about these metrics

Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.

Abstract
High Resolution Image
Download MS PowerPoint Slide
Figure 1
Figure 1. Rational design of 6 and 7 as monovalent C-glycosidic glycomimetic LecB inhibitors. Derivatives of methyl α-d-mannoside 1–5 and their inhibitory potency for the binding with LecB_PAO1. (30, 31, 35) Moieties colored in blue increase potency by improving binding kinetics. (30, 32) The orange colored methyl group originating from l-fucosides enhances binding to LecB through a lipophilic interaction. (31, 35)
High Resolution Image
Download MS PowerPoint Slide
Figure 2
Figure 2. Competitive binding assay of inhibitors with LecB_PAO1 and LecB_PA14. Means and standard deviations were determined from a minimum of three independent measurements. IC₅₀ values for 1 with both LecB variants and 2a–c, 3a,c and 5 with LecB_PAO1 were previously published. (20, 30-32) One representative titration with LecB_PA14 is depicted for reference pairs 2b/6b and 3a/7a. Gray arrows indicate the increase in activity for C-glycosides.
High Resolution Image
Download MS PowerPoint Slide
Scheme 1
Scheme 1. Synthesis of Structures 6a,b and 7a,b^a
High Resolution Image
Download MS PowerPoint Slide
Scheme aReagents and conditions: (a) MeNO₂, DBU, molecular sieves 3 Å, 1,4-dioxane, 50 °C, 3 days; (b) Pt/C, H₂, HCl, MeOH, r.t., 2 days; (c) acyl/sulfonyl chloride or carboxylic acid/EDC·HCl, Et₃N, DMF, 0 °C. Yields are given over two steps from nitro derivative 9.
Figure 3
Figure 3. Isothermal titration microcalorimetry of LecB_PAO1 and LecB_PA14 with ligands 6a and 7a,b. Means and standard deviations were determined from a minimum of three independent titrations. One representative titration graph is depicted for LecB_PAO1 only.
High Resolution Image
Download MS PowerPoint Slide
Figure 4
Figure 4. Surface plasmon resonance (SPR) analysis of direct binding of 6a or 7a to immobilized LecB_PAO1. Experimental data in black; calculated fits using a 1:1 binding model in red.
High Resolution Image
Download MS PowerPoint Slide
Figure 5
Figure 5. Crystal structures of LecB_PA14 with C-glycoside ligands: (A) Complex with trimethylphenyl sulfonamide 7a (1.65 Å resolution), (B) Complex with thiophene 7b (1.65 Å resolution); For the ligands the 2F_obs – F_calc electron density is displayed at 1 σ. Ligands and amino acids of the carbohydrate recognition domain (CRD) are depicted as sticks colored by elements (C: gray, N: blue, O: red, S: yellow); protein surface in transparent blue and two Ca²⁺-ions in the binding sites are shown as green spheres.
High Resolution Image
Download MS PowerPoint Slide
Figure 6
Figure 6. Inhibition of biofilm formation by P. aeruginosa after 48 h growth in the presence of compounds 1, 2a, 3a, 6a, 7a,b, or methyl α-l-fucoside (11) at 100 μM. DMSO in absence of compounds was used as control. (A) Quantification of biofilm biomass. Averages and standard deviations of biofilm formation from three independent replicates. Statistical significance was calculated using the students t test. (B) Raw data of confocal fluorescence microscopy 3D images show one representative z-stack per condition.
High Resolution Image
Download MS PowerPoint Slide
Figure 7
Figure 7. Binding of C-glycosides 6a and 7a and l-fucose (8) to Langerin as determined in a ¹⁹F R₂-filtered NMR competitive binding assay.
High Resolution Image
Download MS PowerPoint Slide
Figure 8
Figure 8. In vivo pharmacokinetics of 7a and 7b in CD-1 mice. Mean profile (±) SD of plasma (left column) and urine (right column) concentration in mouse versus time after i.v. (top) or p.o. (bottom) administration of compound 7a or 7b in a single dosing experiment (10 mg/kg, n = 3). Dashed line represents the in vitro IC₅₀ range for both candidates with LecB_PA14.
High Resolution Image
Download MS PowerPoint Slide
References
This article references 46 other publications.
1. 1
  Nagao, M.; Iinuma, Y.; Igawa, J.; Saito, T.; Yamashita, K.; Kondo, T.; Matsushima, A.; Takakura, S.; Takaori-Kondo, A.; Ichiyama, S. J. Hosp Infect 2011, 79, 49– 53 DOI: 10.1016/j.jhin.2011.04.018
  1
  Control of an outbreak of carbapenem-resistant Pseudomonas aeruginosa in a haemato-oncology unit
  Nagao M; Iinuma Y; Igawa J; Saito T; Yamashita K; Kondo T; Matsushima A; Takakura S; Takaori-Kondo A; Ichiyama S
  The Journal of hospital infection (2011), 79 (1), 49-53 ISSN:.
  An outbreak of a multidrug-resistant Pseudomonas aeruginosa producing metallo-β-lactamase (MBLPA) in a haemato-oncology unit was controlled using multidisciplinary interventions. The present study assesses the effects of these interventions by active surveillance of the incidence of MBLPA infection at the 1,240-bed tertiary care Kyoto University Hospital in Kyoto, Japan. Infection control strategies in 2004 included strengthening contact precautions, analysis of risk factors for MBLPA infection and cessation of urine collection. However, new MBLPA infections were identified in 2006, which prompted enhanced environmental cleaning, routine active surveillance, and restricting carbapenem usage. Between 2004 and 2010, 17 patients in the unit became infected with indistinguishable MBLPA strains. The final five infected patients were found by routine active surveillance, but horizontal transmission was undetectable. The MBLPA outbreak in the haemato-oncology unit was finally contained in 2008.
2. 2
  Rice, L. B. J. Infect. Dis. 2008, 197, 1079– 1081 DOI: 10.1086/533452
  2
  Federal funding for the study of antimicrobial resistance in nosocomial pathogens: no ESKAPE
  Rice Louis B
  The Journal of infectious diseases (2008), 197 (8), 1079-81 ISSN:0022-1899.
  There is no expanded citation for this reference.
3. 3
  Tsutsui, A.; Suzuki, S.; Yamane, K.; Matsui, M.; Konda, T.; Marui, E.; Takahashi, K.; Arakawa, Y. J. Hosp. Infect. 2011, 78, 317– 322 DOI: 10.1016/j.jhin.2011.04.013
  3
  Genotypes and infection sites in an outbreak of multidrug-resistant Pseudomonas aeruginosa
  Tsutsui A; Suzuki S; Yamane K; Matsui M; Konda T; Marui E; Takahashi K; Arakawa Y
  The Journal of hospital infection (2011), 78 (4), 317-22 ISSN:.
  An outbreak of multidrug-resistant (MDR) Pseudomonas aeruginosa occurred in an acute care hospital in Japan, which lasted for more than three years. During January 2006 to June 2009, 59 hospitalised patients with MDR P. aeruginosa were mainly detected by urine culture in the first half, whereas isolation from respiratory tract samples became dominant in the latter half of the outbreak. Non-duplicate MDR P. aeruginosa isolates were available from 51 patients and all isolates were positive for bla(VIM-2). Pulsed-field gel electrophoresis (PFGE) analysis categorised the isolates into three major clusters; types A, B and C with eight, 19 and 21 isolates, respectively. The outbreak started with patients harbouring PFGE type A strains, followed by type B, and type C strains. Multivariate analysis demonstrated that patients with PFGE type C strains were more likely to be detected by respiratory tract samples (odds ratio: 11.87; 95% confidence interval: 1.21-116.86). Improved aseptic urethral catheter care controlled PFGE type A and type B strains and improvement in respiratory care procedures finally contained the transmission of PFGE type C strains.
4. 4
  World Health Organization. WHO publishes list of bacteria for which new antibiotics are urgently needed. http://www.who.int/mediacentre/news/releases/2017/bacteria-antibiotics-needed/en/ (accessed May 2017).
  There is no corresponding record for this reference.
5. 5
  Hauser, A. R.; Rello, J., Eds. Severe Infections Caused by Pseudomonas aeruginosa; Perspectives on Critical Care Infectious Diseases series; Kluwer Academic Publishers Group: Boston, MA, 2003.
  There is no corresponding record for this reference.
6. 6
  Serra, R.; Grande, R.; Butrico, L.; Rossi, A.; Settimio, U. F.; Caroleo, B.; Amato, B.; Gallelli, L.; de Franciscis, S. Expert Rev. Anti-Infect. Ther. 2015, 13, 605– 613 DOI: 10.1586/14787210.2015.1023291
  6
  Chronic wound infections: the role of Pseudomonas aeruginosa and Staphylococcus aureus
  Serra, Raffaele; Grande, Raffaele; Butrico, Lucia; Rossi, Alessio; Settimio, Ugo Francesco; Caroleo, Benedetto; Amato, Bruno; Gallelli, Luca; de Franciscis, Stefano
  Expert Review of Anti-Infective Therapy (2015), 13 (5), 605-613CODEN: ERATCK; ISSN:1478-7210. (Informa Healthcare)
  A review. Chronic leg ulcers affect 1-2% of the general population and are related to increased morbidity and health costs. Staphylococcus aureus and Pseudomonas aeruginosa are the most common bacteria isolated from chronic wounds. They can express virulence factors and surface proteins affecting wound healing. The co-infection of S. aureus and P. aeruginosa is more virulent than single infection. In particular, S. aureus and P. aeruginosa have both intrinsic and acquired antibiotic resistance, making clin. management of infection a real challenge, particularly in patients with comorbidity. Therefore, a correct and prompt diagnosis of chronic wound infection requires a detailed knowledge of skin bacterial flora. This is a necessary prerequisite for tailored pharmacol. treatment, improving symptoms, and reducing side effects and antibiotic resistance.
7. 7
  Mittal, R.; Aggarwal, S.; Sharma, S.; Chhibber, S.; Harjai, K. J. Infect. Public Health 2009, 2, 101– 11 DOI: 10.1016/j.jiph.2009.08.003
  7
  Urinary tract infections caused by Pseudomonas aeruginosa: a minireview
  Mittal Rahul; Aggarwal Sudhir; Sharma Saroj; Chhibber Sanjay; Harjai Kusum
  Journal of infection and public health (2009), 2 (3), 101-11 ISSN:.
  Urinary tract infections (UTIs) are a serious health problem affecting millions of people each year. Infections of the urinary tract are the second most common type of infection in the body. Catheterization of the urinary tract is the most common factor, which predisposes the host to these infections. Catheter-associated UTI (CAUTI) is responsible for 40% of nosocomial infections, making it the most common cause of nosocomial infection. CAUTI accounts for more than 1 million cases in hospitals and nursing homes annually and often involve uropathogens other than Escherichia coli. While the epidemiology and pathogenic mechanisms of uropathogenic Escherichia coli have been extensively studied, little is known about the pathogenesis of UTIs caused by other organisms like Pseudomonas aeruginosa. Scanty available information regarding pathogenesis of UTIs caused by P. aeruginosa is an important bottleneck in developing effective preventive approaches. The aim of this review is to summarize some of the advances made in the field of P. aeruginosa induced UTIs and draws attention of the workers that more basic research at the level of pathogenesis is needed so that novel strategies can be designed.
8. 8
  Poole, K. Front. Microbiol. 2011, 2, 65 DOI: 10.3389/fmicb.2011.00065
  8
  Pseudomonas aeruginosa: resistance to the max
  Poole, Keith
  Frontiers in Cellular and Infection Microbiology (2011), 2 (April), 65CODEN: FCIMAB ISSN:. (Frontiers Media S.A.)
  A review. Pseudomonas aeruginosa is intrinsically resistant to a variety of antimicrobials and can develop resistance during anti-pseudomonal chemotherapy both of which compromise treatment of infections caused by this organism. Resistance to multiple classes of antimicrobials (multidrug resistance) in particular is increasingly common in P. aeruginosa, with a no. of reports of pan-resistant isolates treatable with a single agent, colistin. Acquired resistance in this organism is multifactorial and attributable to chromosomal mutations and the acquisition of resistance genes via horizontal gene transfer. Mutational changes impacting resistance include upregulation of multidrug efflux systems to promote antimicrobial expulsion, derepression of ampC, AmpC alterations that expand the enzyme's substrate specificity (i.e., extended-spectrum AmpC), alterations to outer membrane permeability to limit antimicrobial entry and alterations to antimicrobial targets. Acquired mechanisms contributing to resistance in P. aeruginosa include β-lactamases, notably the extended-spectrum β-lactamases and the carbapenemases that hydrolyze most β-lactams, aminoglycoside-modifying enzymes, and 16S rRNA methylases that provide high-level pan-aminoglycoside resistance. The organism's propensity to grow in vivo as antimicrobial-tolerant biofilms and the occurrence of hypermutator strains that yield antimicrobial resistant mutants at higher frequency also compromise anti-pseudomonal chemotherapy. With limited therapeutic options and increasing resistance will the untreatable P. aeruginosa infection soon be upon us.
9. 9
  Flemming, H.-C.; Wingender, J. Nat. Rev. Microbiol. 2010, 8, 623– 633 DOI: 10.1038/nrmicro2415
  9
  The biofilm matrix
  Flemming, Hans-Curt; Wingender, Jost
  Nature Reviews Microbiology (2010), 8 (9), 623-633CODEN: NRMACK; ISSN:1740-1526. (Nature Publishing Group)
  A review. The microorganisms in biofilms live in a self-produced matrix of hydrated extracellular polymeric substances (EPS) that form their immediate environment. EPS are mainly polysaccharides, proteins, nucleic acids and lipids; they provide the mech. stability of biofilms, mediate their adhesion to surfaces and form a cohesive, three-dimensional polymer network that interconnects and transiently immobilizes biofilm cells. In addn., the biofilm matrix acts as an external digestive system by keeping extracellular enzymes close to the cells, enabling them to metabolize dissolved, colloidal and solid biopolymers. Here we describe the functions, properties and constituents of the EPS matrix that make biofilms the most successful forms of life on earth.
10. 10
  Davies, D. Nat. Rev. Drug Discovery 2003, 2, 114– 122 DOI: 10.1038/nrd1008
  There is no corresponding record for this reference.
11. 11
  Sommer, R.; Joachim, I.; Wagner, S.; Titz, A. Chimia 2013, 67, 286– 290 DOI: 10.2533/chimia.2013.286
  There is no corresponding record for this reference.
12. 12
  Wagner, S.; Sommer, R.; Hinsberger, S.; Lu, C.; Hartmann, R. W.; Empting, M.; Titz, A. J. Med. Chem. 2016, 59, 5929– 5969 DOI: 10.1021/acs.jmedchem.5b01698
  There is no corresponding record for this reference.
13. 13
  Winzer, K.; Falconer, C.; Garber, N. C.; Diggle, S. P.; Camara, M.; Williams, P. J. Bacteriol. 2000, 182, 6401– 6411 DOI: 10.1128/JB.182.22.6401-6411.2000
  13
  The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS
  Winzer, Klaus; Falconer, Colin; Garber, Nachman C.; Diggle, Stephen P.; Camara, Miguel; Williams, Paul
  Journal of Bacteriology (2000), 182 (22), 6401-6411CODEN: JOBAAY; ISSN:0021-9193. (American Society for Microbiology)
  In Pseudomonas aeruginosa, many exoproduct virulence determinants are regulated via a hierarchical quorum-sensing cascade involving the transcriptional regulators LasR and RhlR and their cognate activators, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL). In this paper, we demonstrate that the cytotoxic lectins PA-IL and PA-IIL are regulated via quorum sensing. Using immunoblot anal., the prodn. of both lectins was found to be directly dependent on the rhl locus while, in a lasR mutant, the onset of lectin synthesis was delayed but not abolished. The PA-IL structural gene, lecA, was cloned and sequenced. Transcript anal. indicated a monocistronic organization with a transcriptional start site 70 bp upstream of the lecA translational start codon. A lux box-type element together with RpoS (σS) consensus sequences was identified upstream of the putative promoter region. In Escherichia coli, expression of a lecA::lux reporter fusion was activated by RhlR/C4-HSL, but not by LasR/3O-C12-HSL, confirming direct regulation by RhlR/C4-HSL. Similarly, in P. aeruginosa PAO1, the expression of a chromosomal lecA::lux fusion was enhanced but not advanced by the addn. of exogenous C4-HSL but not 3O-C12-HSL. Furthermore, mutation of rpoS abolished lectin synthesis in P. aeruginosa, demonstrating that both RpoS and RhlR/C4-HSL are required. Although the C4-HSL-dependent expression of the lecA::lux reporter in E. coli could be inhibited by the presence of 3O-C12-HSL, this did not occur in P. aeruginosa. This suggests that, in the homologous genetic background, 3O-C12-HSL does not function as a posttranslational regulator of the RhlR/C4-HSL-dependent activation of lecA expression.
14. 14
  Diggle, S. P.; Stacey, R. E.; Dodd, C.; Cámara, M.; Williams, P.; Winzer, K. Environ. Microbiol. 2006, 8, 1095– 1104 DOI: 10.1111/j.1462-2920.2006.001001.x
  There is no corresponding record for this reference.
15. 15
  Tielker, D.; Hacker, S.; Loris, R.; Strathmann, M.; Wingender, J.; Wilhelm, S.; Rosenau, F.; Jaeger, K.-E. Microbiology 2005, 151, 1313– 1323 DOI: 10.1099/mic.0.27701-0
  There is no corresponding record for this reference.
16. 16
  Gilboa-Garber, N. Methods Enzymol. 1982, 83, 378– 385 DOI: 10.1016/0076-6879(82)83034-6
  There is no corresponding record for this reference.
17. 17
  Klockgether, J.; Cramer, N.; Wiehlmann, L.; Davenport, C. F.; Tümmler, B. Front. Microbiol. 2011, 2, 150 DOI: 10.3389/fmicb.2011.00150
  17
  Pseudomonas aeruginosa genomic structure and diversity
  Klockgether, Jens; Cramer, Nina; Wiehlmann, Lutz; Davenport, Colin F.; Tuemmler, Burkhard
  Frontiers in Cellular and Infection Microbiology (2011), 2 (July), 150CODEN: FCIMAB ISSN:. (Frontiers Media S.A.)
  A review. The Pseudomonas aeruginosa genome (G+C content 65-67%, size 5.5-7 Mbp) is made up of a single circular chromosome and a variable no. of plasmids. Sequencing of complete genomes or blocks of the accessory genome has revealed that the genome encodes a large repertoire of transporters, transcriptional regulators and two-component regulatory systems which reflects its metabolic diversity to utilize a broad range of nutrients. The conserved core component of the genome is largely collinear among P. aeruginosa strains and exhibits an interclonal sequence diversity of 0.5-0.7 %. Only a few loci of the core genome are subject to diversifying selection. Genome diversity is mainly caused by accessory DNA elements located in 79 regions of genome plasticity that are scattered around the genome and show an anomalous usage of mono- to tetradecanucleotides. Genomic islands of the pKLC102/PAGI-2 family that integrate into tRNALys or tRNAGly genes represent hotspots of inter- and intra-clonal genomic diversity. The individual islands differ in their repertoire of metabolic genes that make a large contribution to the pangenome. In order to unravel intraclonal diversity of P. aeruginosa, the genomes of two members of the PA14 clonal complex from diverse habitats and geog. origin were compared. The genome sequences differed by less than 0.01% from each other. 198 Of the 231 SNPs were non-randomly distributed in the genome. Non-synonymous SNPs were mainly found in an integrated Pf1-like phage and in genes involved in transcriptional regulation, membrane and extracellular constituents, transport and secretion. In summary, P. aeruginosa is endowed with a highly conserved core genome of low sequence diversity and a highly variable accessory genome that communicates with other pseudomonads and genera via horizontal gene transfer.
18. 18
  Dötsch, A.; Schniederjans, M.; Khaledi, A.; Hornischer, K.; Schulz, S.; Bielecka, A.; Eckweiler, D.; Pohl, S.; Häussler, S. mBio 2015, 6, e00749-15 DOI: 10.1128/mBio.00749-15
  There is no corresponding record for this reference.
19. 19
  Boukerb, A. M.; Decor, A.; Ribun, S.; Tabaroni, R.; Rousset, A.; Commin, L.; Buff, S.; Doléans-Jordheim, A.; Vidal, S.; Varrot, A.; Imberty, A.; Cournoyer, B. Front. Microbiol. 2016, 7, 811 DOI: 10.3389/fmicb.2016.00811
  19
  Genomic Rearrangements and Functional Diversification of lecA and lecB Lectin-Coding Regions Impacting the Efficacy of Glycomimetics Directed against Pseudomonas aeruginosa
  Boukerb Amine M; Ribun Sebastien; Doleans-Jordheim Anne; Cournoyer Benoit; Decor Aude; Tabaroni Rachel; Varrot Annabelle; Imberty Anne; Rousset Audric; Vidal Sebastien; Commin Loris; Buff Samuel
  Frontiers in microbiology (2016), 7 (), 811 ISSN:1664-302X.
  LecA and LecB tetrameric lectins take part in oligosaccharide-mediated adhesion-processes of Pseudomonas aeruginosa. Glycomimetics have been designed to block these interactions. The great versatility of P. aeruginosa suggests that the range of application of these glycomimetics could be restricted to genotypes with particular lectin types. The likelihood of having genomic and genetic changes impacting LecA and LecB interactions with glycomimetics such as galactosylated and fucosylated calix[4]arene was investigated over a collection of strains from the main clades of P. aeruginosa. Lectin types were defined, and their ligand specificities were inferred. These analyses showed a loss of lecA among the PA7 clade. Genomic changes impacting lec loci were thus assessed using strains of this clade, and by making comparisons with the PAO1 genome. The lecA regions were found challenged by phage attacks and PAGI-2 (genomic island) integrations. A prophage was linked to the loss of lecA. The lecB regions were found less impacted by such rearrangements but greater lecB than lecA genetic divergences were recorded. Sixteen combinations of LecA and LecB types were observed. Amino acid variations were mapped on PAO1 crystal structures. Most significant changes were observed on LecBPA7, and found close to the fucose binding site. Glycan array analyses were performed with purified LecBPA7. LecBPA7 was found less specific for fucosylated oligosaccharides than LecBPAO1, with a preference for H type 2 rather than type 1, and Lewis(a) rather than Lewis(x). Comparison of the crystal structures of LecBPA7 and LecBPAO1 in complex with Lewis(a) showed these changes in specificity to have resulted from a modification of the water network between the lectin, galactose and GlcNAc residues. Incidence of these modifications on the interactions with calix[4]arene glycomimetics at the cell level was investigated. An aggregation test was used to establish the efficacy of these ligands. Great variations in the responses were observed. Glycomimetics directed against LecB yielded the highest numbers of aggregates for strains from all clades. The use of a PAO1ΔlecB strain confirmed a role of LecB in this aggregation phenotype. Fucosylated calix[4]arene showed the greatest potential for a use in the prevention of P. aeruginosa infections.
20. 20
  Sommer, R.; Wagner, S.; Varrot, A.; Nycholat, C. M.; Khaledi, A.; Haussler, S.; Paulson, J. C.; Imberty, A.; Titz, A. Chem. Sci. 2016, 7, 4990– 5001 DOI: 10.1039/C6SC00696E
  20
  The virulence factor LecB varies in clinical isolates: consequences for ligand binding and drug discovery
  Sommer, Roman; Wagner, Stefanie; Varrot, Annabelle; Nycholat, Corwin M.; Khaledi, Ariane; Haeussler, Susanne; Paulson, James C.; Imberty, Anne; Titz, Alexander
  Chemical Science (2016), 7 (8), 4990-5001CODEN: CSHCCN; ISSN:2041-6520. (Royal Society of Chemistry)
  P. aeruginosa causes a substantial no. of nosocomial infections and is the leading cause of death of cystic fibrosis patients. This Gram-neg. bacterium is highly resistant against antibiotics and further protects itself by forming a biofilm. Moreover, a high genomic variability among clin. isolates complicates therapy. Its lectin LecB is a virulence factor and necessary for adhesion and biofilm formation. The authors analyzed the sequence of LecB variants in a library of clin. isolates and demonstrate that it can serve as a marker for strain family classification. LecB from the highly virulent model strain PA14 presents 13% sequence divergence with LecB from the well characterized PAO1 strain. These differences might result in differing ligand binding specificities and ultimately in reduced efficacy of drugs directed towards LecB. Despite several amino acid variations at the carbohydrate binding site, glycan array anal. showed a comparable binding pattern for both variants. A common high affinity ligand could be identified and after its chemoenzymic synthesis verified in a competitive binding assay: an N-glycan presenting two blood group O epitopes (H-type 2 antigen). Mol. modeling of the complex suggests a bivalent interaction of the ligand with the LecB tetramer by bridging two sep. binding sites. This binding rationalizes the strong avidity (35 nM) of LecBPA14 to this human fucosylated N-glycan. Biochem. evaluation of a panel of glycan ligands revealed that LecBPA14 demonstrated higher glycan affinity compared to LecBPAO1 including the extraordinarily potent affinity of 70 nM towards the monovalent human antigen Lewisa. The structural basis of this unusual high affinity ligand binding for lectins was rationalized by solving the protein crystal structures of LecBPA14 with several glycans.
21. 21
  Boukerb, A. M.; Rousset, A.; Galanos, N.; Méar, J.-B.; Thepaut, M.; Grandjean, T.; Gillon, E.; Cecioni, S.; Abderrahmen, C.; Faure, K.; Redelberger, D.; Kipnis, E.; Dessein, R.; Havet, S.; Darblade, B.; Matthews, S. E.; de Bentzmann, S.; Guéry, B.; Cournoyer, B.; Imberty, A.; Vidal, S. J. Med. Chem. 2014, 57, 10275– 10289 DOI: 10.1021/jm500038p
  There is no corresponding record for this reference.
22. 22
  Chemani, C.; Imberty, A.; de Bentzmann, S.; Pierre, M.; Wimmerová, M.; Guery, B. P.; Faure, K. Infect. Immun. 2009, 77, 2065– 2075 DOI: 10.1128/IAI.01204-08
  22
  Role of LecA and LecB lectins in Pseudomonas aeruginosa-induced lung injury and effect of carbohydrate ligands
  Chemani, Chanez; Imberty, Anne; de Bentzmann, Sophie; Pierre, Maud; Wimmerova, Michaela; Guery, Benoit P.; Faure, Karine
  Infection and Immunity (2009), 77 (5), 2065-2075CODEN: INFIBR; ISSN:0019-9567. (American Society for Microbiology)
  Pseudomonas aeruginosa is a frequently encountered pathogen that is involved in acute and chronic lung infections. Lectin-mediated bacterium-cell recognition and adhesion are crit. steps in initiating P. aeruginosa pathogenesis. This study was designed to evaluate the contributions of LecA and LecB to the pathogenesis of P. aeruginosa-mediated acute lung injury. Using an in vitro model with A549 cells and an exptl. in vivo murine model of acute lung injury, we compared the parental strain to lecA and lecB mutants. The effects of both LecA- and LecB-specific lectin-inhibiting carbohydrates (α-methyl-galactoside and α-methyl-fucoside, resp.) were evaluated. In vitro, the parental strain was assocd. with increased cytotoxicity and adhesion on A549 cells compared to the lecA and lecB mutants. In vivo, the P. aeruginosa-induced increase in alveolar barrier permeability was reduced with both mutants. The bacterial burden and dissemination were decreased for both mutants compared with the parental strain. Coadministration of specific lectin inhibitors markedly reduced lung injury and mortality. Our results demonstrate that there is a relationship between lectins and the pathogenicity of P. aeruginosa. Inhibition of the lectins by specific carbohydrates may provide new therapeutic perspectives.
23. 23
  Cott, C.; Thuenauer, R.; Landi, A.; Kühn, K.; Juillot, S.; Imberty, A.; Madl, J.; Eierhoff, T.; Römer, W. Biochim. Biophys. Acta, Mol. Cell Res. 2016, 1863, 1106– 1118 DOI: 10.1016/j.bbamcr.2016.02.004
  23
  Pseudomonas aeruginosa lectin LecB inhibits tissue repair processes by triggering β-catenin degradation
  Cott, Catherine; Thuenauer, Roland; Landi, Alessia; Kuehn, Katja; Juillot, Samuel; Imberty, Anne; Madl, Josef; Eierhoff, Thorsten; Roemer, Winfried
  Biochimica et Biophysica Acta, Molecular Cell Research (2016), 1863 (6_Part_A), 1106-1118CODEN: BBAMCO; ISSN:0167-4889. (Elsevier B.V.)
  Pseudomonas aeruginosa is an opportunistic pathogen that induces severe lung infections such as ventilator-assocd. pneumonia and acute lung injury. Under these conditions, the bacterium diminishes epithelial integrity and inhibits tissue repair mechanisms, leading to persistent infections. Understanding the involved bacterial virulence factors and their mode of action is essential for the development of new therapeutic approaches. In our study we discovered a so far unknown effect of the P. aeruginosa lectin LecB on host cell physiol. LecB alone was sufficient to attenuate migration and proliferation of human lung epithelial cells and to induce transcriptional activity of NF-κB. These effects are characteristic of impaired tissue repair. Moreover, we found a strong degrdn. of β-catenin, which was partially recovered by the proteasome inhibitor lactacystin. In addn., LecB induced loss of cell-cell contacts and reduced expression of the β-catenin targets c-myc and cyclin D1. Blocking of LecB binding to host cell plasma membrane receptors by sol. L-fucose prevented these changes in host cell behavior and signaling, and thereby provides a powerful strategy to suppress LecB function. Our findings suggest that P. aeruginosa employs LecB as a virulence factor to induce β-catenin degrdn., which then represses processes that are directly linked to tissue recovery.
24. 24
  Mitchell, E.; Houles, C.; Sudakevitz, D.; Wimmerova, M.; Gautier, C.; Pérez, S.; Wu, A. M.; Gilboa-Garber, N.; Imberty, A. Nat. Struct. Biol. 2002, 9, 918– 921 DOI: 10.1038/nsb865
  There is no corresponding record for this reference.
25. 25
  Hauber, H.-P.; Schulz, M.; Pforte, A.; Mack, D.; Zabel, P.; Schumacher, U. Int. J. Med. Sci. 2008, 5, 371– 376 DOI: 10.7150/ijms.5.371
  25
  Inhalation with fucose and galactose for treatment of Pseudomonas aeruginosa in cystic fibrosis patients
  Hauber, Hans-Peter; Schulz, Maria; Pforte, Almuth; Mack, Dietrich; Zabel, Peter; Schumacher, Udo
  International Journal of Medical Sciences (2008), 5 (6), 371-376CODEN: IJMSGZ; ISSN:1449-1907. (Ivyspring International Publisher)
  Background: Colonisation of cystic fibrosis (CF) lungs with Pseudomonas aeruginosa is facilitated by two lectins, which bind to the sugar coat of the surface lining epithelia and stop the cilia beating. Objectives: We hypothesized that P. aeruginosa lung infection should be cleared by inhalation of fucose and galactose, which compete for the sugar binding site of the two lectins and thus inhibit the binding of P. aeruginosa. Methods: 11 adult CF patients with chronic infection with P. aeruginosa were treated twice daily with inhalation of a fucose/galactose soln. for 21 days (4 patients only received inhalation, 7 patients received inhalation and i.v. antibiotics). Microbial counts of P. aeruginosa, lung function measurements, and inflammatory markers were detd. before and after treatment. Results: The sugar inhalation was well tolerated and no adverse side effects were obsd. Inhalation alone as well as combined therapy (inhalation and antibiotics) significantly decreased P. aeruginosa in sputum (P < 0.05). Both therapies also significantly reduced TNFα expression in sputum and peripheral blood cells (P < 0.05). No change in lung function measurements was obsd. Conclusions: Inhalation of simple sugars is a safe and effective measure to reduce the P. aeruginosa counts in CF patients. This may provide an alternative therapeutical approach to treat infection with P. aeruginosa.
26. 26
  Bucior, I.; Abbott, J.; Song, Y.; Matthay, M. A.; Engel, J. N. Am. J. Physiol Lung Cell Mol. Physiol 2013, 305, L352– 363 DOI: 10.1152/ajplung.00387.2012
  26
  Sugar administration is an effective adjunctive therapy in the treatment of Pseudomonas aeruginosa pneumonia
  Bucior, Iwona; Abbott, Jason; Song, Yuanlin; Matthay, Michael A.; Engel, Joanne N.
  American Journal of Physiology (2013), 305 (3, Pt. 1), L352-L363CODEN: AJPHAP; ISSN:0002-9513. (American Physiological Society)
  Treatment of acute and chronic pulmonary infections caused by opportunistic pathogen Pseudomonas aeruginosa is limited by the increasing frequency of multidrug bacterial resistance. Here, we describe a novel adjunctive therapy in which administration of a mix of simple sugars-mannose, fucose, and galactose-inhibits bacterial attachment, limits lung damage, and potentiates conventional antibiotic therapy. The sugar mixt. inhibits adhesion of nonmucoid and mucoid P. aeruginosa strains to bronchial epithelial cells in vitro. In a murine model of acute pneumonia, treatment with the sugar mixt. alone diminishes lung damage, bacterial dissemination to the subpleural alveoli, and neutrophil- and IL-8-driven inflammatory responses. Remarkably, the sugars act synergistically with anti-Pseudomonas antibiotics, including β-lactams and quinolones, to further reduce bacterial lung colonization and damage. To probe the mechanism, we examd. the effects of sugars in the presence or absence of antibiotics during growth in liq. culture and in an ex vivo infection model utilizing freshly dissected mouse tracheas and lungs. We demonstrate that the sugar mixt. induces rapid but reversible formation of bacterial clusters that exhibited enhanced susceptibility to antibiotics compared with individual bacteria. Our findings reveal that sugar inhalation, an inexpensive and safe therapeutic, could be used in combination with conventional antibiotic therapy to more effectively treat P. aeruginosa lung infections.
27. 27
  Cecioni, S.; Imberty, A.; Vidal, S. Chem. Rev. 2015, 115, 525– 561 DOI: 10.1021/cr500303t
  There is no corresponding record for this reference.
28. 28
  Bernardi, A.; Jiménez-Barbero, J.; Casnati, A.; De Castro, C.; Darbre, T.; Fieschi, F.; Finne, J.; Funken, H.; Jaeger, K.-E.; Lahmann, M.; Lindhorst, T. K.; Marradi, M.; Messner, P.; Molinaro, A.; Murphy, P. V.; Nativi, C.; Oscarson, S.; Penadés, S.; Peri, F.; Pieters, R. J.; Renaudet, O.; Reymond, J.-L.; Richichi, B.; Rojo, J.; Sansone, F.; Schäffer, C.; Turnbull, W. B.; Velasco-Torrijos, T.; Vidal, S.; Vincent, S.; Wennekes, T.; Zuilhof, H.; Imberty, A. Chem. Soc. Rev. 2013, 42, 4709– 4727 DOI: 10.1039/C2CS35408J
  There is no corresponding record for this reference.
29. 29
  Johansson, E. M. V.; Crusz, S. A.; Kolomiets, E.; Buts, L.; Kadam, R. U.; Cacciarini, M.; Bartels, K.-M.; Diggle, S. P.; Cámara, M.; Williams, P.; Loris, R.; Nativi, C.; Rosenau, F.; Jaeger, K.-E.; Darbre, T.; Reymond, J.-L. Chem. Biol. 2008, 15, 1249– 1257 DOI: 10.1016/j.chembiol.2008.10.009
  29
  Inhibition and Dispersion of Pseudomonas aeruginosa Biofilms by Glycopeptide Dendrimers Targeting the Fucose-Specific Lectin LecB
  Johansson, Emma M. V.; Crusz, Shanika A.; Kolomiets, Elena; Buts, Lieven; Kadam, Rameshwar U.; Cacciarini, Martina; Bartels, Kai-Malte; Diggle, Stephen P.; Camara, Miguel; Williams, Paul; Loris, Remy; Nativi, Cristina; Rosenau, Frank; Jaeger, Karl-Erich; Darbre, Tamis; Reymond, Jean-Louis
  Chemistry & Biology (Cambridge, MA, United States) (2008), 15 (12), 1249-1257CODEN: CBOLE2; ISSN:1074-5521. (Cell Press)
  The human pathogenic bacterium Pseudomonas aeruginosa produces a fucose-specific lectin, LecB, implicated in tissue attachment and the formation of biofilms. To investigate if LecB inhibition disrupts these processes, high-affinity ligands were obtained by screening two 15,536-member combinatorial libraries of multivalent fucosyl-peptide dendrimers. The most potent LecB-ligands identified were dendrimers FD2 (C-Fuc-LysProLeu)4(LysPheLysIle)2 LysHisIleNH2 (IC50 = 0.14 μM by ELLA) and PA8 (OFuc-LysAlaAsp)4(LysSerGlyAla)2 LysHisIleNH2 (IC50 = 0.11 μM by ELLA). Dendrimer FD2 led to complete inhibition of P. aeruginosa biofilm formation (IC50 ∼ 10 μM) and induced complete dispersion of established biofilms in the wild-type strain and in several clin. P. aeruginosa isolates. These expts. suggest that LecB inhibition by high-affinity multivalent ligands could represent a therapeutic approach against P. aeruginosa infections by inhibition of biofilm formation and dispersion of established biofilms.
30. 30
  Hauck, D.; Joachim, I.; Frommeyer, B.; Varrot, A.; Philipp, B.; Möller, H. M.; Imberty, A.; Exner, T. E.; Titz, A. ACS Chem. Biol. 2013, 8, 1775– 1784 DOI: 10.1021/cb400371r
  There is no corresponding record for this reference.
31. 31
  Sommer, R.; Exner, T. E.; Titz, A. PLoS One 2014, 9, e112822 DOI: 10.1371/journal.pone.0112822
  There is no corresponding record for this reference.
32. 32
  Sommer, R.; Hauck, D.; Varrot, A.; Wagner, S.; Audfray, A.; Prestel, A.; Möller, H. M.; Imberty, A.; Titz, A. ChemistryOpen 2015, 4, 756– 767 DOI: 10.1002/open.201500162
  There is no corresponding record for this reference.
33. 33
  Hofmann, A.; Sommer, R.; Hauck, D.; Stifel, J.; Göttker-Schnetmann, I.; Titz, A. Carbohydr. Res. 2015, 412, 34– 42 DOI: 10.1016/j.carres.2015.04.010
  There is no corresponding record for this reference.
34. 34
  Beshr, G.; Sommer, R.; Hauck, D.; Siebert, D. C. B.; Hofmann, A.; Imberty, A.; Titz, A. MedChemComm 2016, 7, 519– 530 DOI: 10.1039/C5MD00557D
  There is no corresponding record for this reference.
35. 35
  Sabin, C.; Mitchell, E. P.; Pokorná, M.; Gautier, C.; Utille, J.-P.; Wimmerová, M.; Imberty, A. FEBS Lett. 2006, 580, 982– 987 DOI: 10.1016/j.febslet.2006.01.030
  There is no corresponding record for this reference.
36. 36
  Tarcsay, A.; Keseru, G. M. Drug Discovery Today 2015, 20, 86– 94 DOI: 10.1016/j.drudis.2014.09.014
  36
  Is there a link between selectivity and binding thermodynamics profiles?
  Tarcsay, Akos; Keseru, Gyorgy M.
  Drug Discovery Today (2015), 20 (1), 86-94CODEN: DDTOFS; ISSN:1359-6446. (Elsevier Ltd.)
  A review. Thermodn. of ligand binding is influenced by the interplay between enthalpy and entropy contributions of the binding event. The impact of these binding free energy components, however, is not limited to the primary target only. Here, we investigate the relationship between binding thermodn. and selectivity profiles by combining publicly available data from broad off-target assay profiling and the corresponding thermodn. measurements. Our anal. indicates that compds. binding their primary targets with higher entropy contributions tend to hit more off-targets compared with those ligands that demonstrated enthalpy-driven binding.
37. 37
  Hopkins, A. L.; Keseru, G. M.; Leeson, P. D.; Rees, D. C.; Reynolds, C. H. Nat. Rev. Drug Discovery 2014, 13, 105– 121 DOI: 10.1038/nrd4163
  37
  The role of ligand efficiency metrics in drug discovery
  Hopkins, Andrew L.; Keserue, Gyoergy M.; Leeson, Paul D.; Rees, David C.; Reynolds, Charles H.
  Nature Reviews Drug Discovery (2014), 13 (2), 105-121CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)
  A review. The judicious application of ligand or binding efficiency metrics, which quantify the mol. properties required to obtain binding affinity for a drug target, is gaining traction in the selection and optimization of fragments, hits and leads. Retrospective anal. of recently marketed oral drugs shows that they frequently have highly optimized ligand efficiency values for their targets. Optimizing ligand efficiency metrics based on both mol. mass and lipophilicity, when set in the context of the specific target, has the potential to ameliorate the inflation of these properties that has been obsd. in current medicinal chem. practice, and to increase the quality of drug candidates.
38. 38
  Tummino, P. J.; Copeland, R. A. Biochemistry 2008, 47, 5481– 5492 DOI: 10.1021/bi8002023
  There is no corresponding record for this reference.
39. 39
  Perret, S.; Sabin, C.; Dumon, C.; Pokorná, M.; Gautier, C.; Galanina, O.; Ilia, S.; Bovin, N.; Nicaise, M.; Desmadril, M.; Gilboa-Garber, N.; Wimmerová, M.; Mitchell, E. P.; Imberty, A. Biochem. J. 2005, 389, 325– 332 DOI: 10.1042/BJ20050079
  There is no corresponding record for this reference.
40. 40
  Loris, R.; Tielker, D.; Jaeger, K.-E.; Wyns, L. J. Mol. Biol. 2003, 331, 861– 870 DOI: 10.1016/S0022-2836(03)00754-X
  There is no corresponding record for this reference.
41. 41
  Lagendijk, E. L.; Validov, S.; Lamers, G. E. M.; de Weert, S.; Bloemberg, G. V. FEMS Microbiol. Lett. 2010, 305, 81– 90 DOI: 10.1111/j.1574-6968.2010.01916.x
  There is no corresponding record for this reference.
42. 42
  Mayer, S.; Raulf, M.-K.; Lepenies, B. Histochem. Cell Biol. 2017, 147, 223– 237 DOI: 10.1007/s00418-016-1523-7
  42
  C-type lectins: their network and roles in pathogen recognition and immunity
  Mayer, Sabine; Raulf, Marie-Kristin; Lepenies, Bernd
  Histochemistry and Cell Biology (2017), 147 (2), 223-237CODEN: HCBIFP; ISSN:0948-6143. (Springer)
  C-type lectins (CTLs) represent the most complex family of animal/human lectins that comprises 17 different groups. During evolution, CTLs have developed by diversification to cover a broad range of glycan ligands. However, ligand binding by CTLs is not necessarily restricted to glycans as some CTLs also bind to proteins, lipids, inorg. mols., or ice crystals. CTLs share a common fold that harbors a Ca2+ for contact to the sugar and about 18 invariant residues in a phylogenetically conserved pattern. In vertebrates, CTLs have numerous functions, including serum glycoprotein homeostasis, pathogen sensing, and the initiation of immune responses. Myeloid CTLs in innate immunity are mainly expressed by antigen-presenting cells and play a prominent role in the recognition of a variety of pathogens such as fungi, bacteria, viruses, and parasites. However, myeloid CTLs such as the macrophage inducible CTL (Mincle) or Clec-9a may also bind to self-antigens and thus contribute to immune homeostasis. While some CTLs induce pro-inflammatory responses and thereby lead to activation of adaptive immune responses, other CTLs act as inhibitory receptors and dampen cellular functions. Since CTLs are key players in pathogen recognition and innate immunity, targeting CTLs may be a promising strategy for cell-specific delivery of drugs or vaccine antigens and to modulate immune responses.
43. 43
  Holla, A.; Skerra, A. Protein Eng., Des. Sel. 2011, 24, 659– 669 DOI: 10.1093/protein/gzr016
  43
  Comparative analysis reveals selective recognition of glycans by the dendritic cell receptors DC-SIGN and Langerin
  Holla, Andrea; Skerra, Arne
  Protein Engineering, Design & Selection (2011), 24 (9), 659-669CODEN: PEDSBR; ISSN:1741-0126. (Oxford University Press)
  DC-SIGN (dendritic cell-specific ICAM-3 grabbing non-integrin) and Langerin are homologous C-type lectins expressed as cell-surface receptors on different populations of dendritic cells (DCs). DC-SIGN interacts with glycan structures on HIV-1, facilitating virus survival, transmission and infection, whereas Langerin, which is characteristic of Langerhans cells (LCs), promotes HIV-1 uptake and degrdn. Here we describe a comprehensive comparison of the glycan specificities of both proteins by probing a synthetic carbohydrate microarray comprising 275 sugar compds. using the bacterially produced and fluorescence-labeled, monomeric carbohydrate-recognition domains (CRDs) of DC-SIGN and Langerin. In this side-by-side study DC-SIGN was found to preferentially bind internal mannose residues of high-mannose-type saccharides and the fucose-contg. blood-type antigens H, A, B, Lea, Leb Lex, Ley, sialyl-Lea as well as sulfatated derivs. of Lea and Lex. In contrast, Langerin appeared to recognize a different spectrum of compds., esp. those contg. terminal mannose, terminal N-acetylglucosamine and 6-sulfogalactose residues, but also the blood-type antigens H, A and B. Of the Lewis antigens, only Leb, Ley, sialyl-Lea and the sialyl-Lex deriv. with 6'-sulfatation at the galactose (sialyl-6SGal Lex) were weakly bound by Langerin. Notably, Ca2+-independent glycan-binding activity of Langerin could not be detected either by probing the glycan array or by isothermal titrn. calorimetry of the CRD with mannose and mannobiose. The precise knowledge of carbohydrate specificity of DC-SIGN and Langerin receptors resulting from our study may aid the future design of microbicides that specifically affect the DC-SIGN/HIV-1 interaction while not compromising the protective function of Langerin.
44. 44
  Wamhoff, E.-C.; Hanske, J.; Schnirch, L.; Aretz, J.; Grube, M.; Varón Silva, D.; Rademacher, C. ACS Chem. Biol. 2016, 11, 2407– 2413 DOI: 10.1021/acschembio.6b00561
  There is no corresponding record for this reference.
45. 45
  Zhang, Y.; Huo, M.; Zhou, J.; Xie, S. Comput. Methods Programs Biomed 2010, 99, 306– 314 DOI: 10.1016/j.cmpb.2010.01.007
  45
  PKSolver: An add-in program for pharmacokinetic and pharmacodynamic data analysis in Microsoft Excel
  Zhang Yong; Huo Meirong; Zhou Jianping; Xie Shaofei
  Computer methods and programs in biomedicine (2010), 99 (3), 306-14 ISSN:.
  This study presents PKSolver, a freely available menu-driven add-in program for Microsoft Excel written in Visual Basic for Applications (VBA), for solving basic problems in pharmacokinetic (PK) and pharmacodynamic (PD) data analysis. The program provides a range of modules for PK and PD analysis including noncompartmental analysis (NCA), compartmental analysis (CA), and pharmacodynamic modeling. Two special built-in modules, multiple absorption sites (MAS) and enterohepatic circulation (EHC), were developed for fitting the double-peak concentration-time profile based on the classical one-compartment model. In addition, twenty frequently used pharmacokinetic functions were encoded as a macro and can be directly accessed in an Excel spreadsheet. To evaluate the program, a detailed comparison of modeling PK data using PKSolver and professional PK/PD software package WinNonlin and Scientist was performed. The results showed that the parameters estimated with PKSolver were satisfactory. In conclusion, the PKSolver simplified the PK and PD data analysis process and its output could be generated in Microsoft Word in the form of an integrated report. The program provides pharmacokinetic researchers with a fast and easy-to-use tool for routine and basic PK and PD data analysis with a more user-friendly interface.
46. 46
  Allen, R. C.; Popat, R.; Diggle, S. P.; Brown, S. P. Nat. Rev. Microbiol. 2014, 12, 300– 308 DOI: 10.1038/nrmicro3232
  46
  Targeting virulence: can we make evolution-proof drugs?
  Allen, Richard C.; Popat, Roman; Diggle, Stephen P.; Brown, Sam P.
  Nature Reviews Microbiology (2014), 12 (4), 300-308CODEN: NRMACK; ISSN:1740-1526. (Nature Publishing Group)
  A review and discussion. Antivirulence drugs are a new type of therapeutic drug that target virulence factors, potentially revitalising the drug-development pipeline with new targets. As antivirulence drugs disarm the pathogen, rather than kill or halt pathogen growth, it has been hypothesized that they will generate much weaker selection for resistance than traditional antibiotics. However, recent studies have shown that mechanisms of resistance to antivirulence drugs exist, seemingly damaging the 'evolution-proof' claim. In this Opinion article, we highlight a crucial distinction between whether resistance can emerge and whether it will spread to a high frequency under drug selection. We argue that selection for resistance can be reduced, or even reversed, using appropriate combinations of target and treatment environment, opening a path towards the development of evolutionarily robust novel therapeutics.
- PDB: 1OUR
- PDB: 5A6Y
Supporting Information
Supporting Information
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/jacs.7b11133.
- Experimental details and ¹H and ¹³C spectra of new compounds; ITC titration data; X-ray data collection and refinement statistics; crystal structures showing ligand alignment and crystal packing effects; correlation of fluorescence intensities with cfu and OD600 measurements; effects of compounds on total fluorescence intensities; calculated lipophilicity of selected compounds; analysis of TNF-α concentration after stimulation of mouse spleen cells with and without test compounds; microsomal intrinsic clearance (CL_int) of 3a, 3b, and C-glycosides 7 in mouse and human liver microsomes; stability of LecB ligands in mouse plasma; m/z search window for plasma stability assay; toxicity of LecB ligands to human liver Hep G2 cells; accuracy, quantification limits, and lower limit of qualification for 7a and 7b in plasma, urine, and kidney matrix; mass spectrometric conditions used for quantification and qualification of 7a, 7b, and the internal standard glipizide; PK parameters of 7a and 7b in mice (PDF)
- ja7b11133_si_001.pdf (4.35 MB)
Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Glycomimetic, Orally Bioavailable LecB Inhibitors Block Biofilm Formation of Pseudomonas aeruginosaClick to copy article linkArticle link copied!

Journal of the American Chemical Society

Abstract

Note Added after ASAP Publication

Introduction

Results and Discussion

Design and Synthesis of C-Glycoside LecB Inhibitors

Figure 1

Figure 2

Scheme 1

Improved Binding Properties of C-Glycoside Structures toward LecB

Figure 3

Figure 4

Structure of LecB in Complex with Hybrid Inhibitors

Figure 5

Low Molecular Weight Glycomimetics Are Potent Inhibitors of Bacterial Biofilm Formation

Figure 6

Compound Selectivity for LecB over Host Lectins

Figure 7

In Vitro Metabolic Stability and Toxicity

In Vivo Pharmacokinetics of C-Glycosides

Figure 8

Conclusions

Supporting Information

Terms & Conditions

Author Information

Acknowledgment

References

Cited By

Journal of the American Chemical Society

Article Views

Altmetric

Citations

Recommended Articles

Abstract

Figure 1

Figure 2

Scheme 1

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

References

Supporting Information

Supporting Information

Terms & Conditions

Glycomimetic, Orally Bioavailable LecB Inhibitors Block Biofilm Formation of Pseudomonas aeruginosa
Click to copy article linkArticle link copied!