Discovery of quinazoline HPK1 inhibitors with high cellular potency.

Hematopoietic progenitor kinase 1 (HPK1) is regarded as a highly validated target in pre-clinical immune oncology. HPK1 has been described as regulating multiple critical signaling pathway in both adaptive and innate cells. In support of this role, HPK1 KO T cells show enhanced sensitivity to TCR activation and HPK1 KO mice display enhanced anti-tumor activity. Taken together, inhibition of HPK1 has the potential to induce enhanced anti-tumor immune response. Herein, we described the discovery of highly potent HPK1 inhibitors starting form a weak HTS hit. Using a structure-based drug design, HPK1 inhibitors exhibiting excellent cellular single-digit nanomolar potency in both proximal (pSLP76) and distal (IL-2) biomarkers along with sustained elevation of IL-2 cytokine secretion were discovered.

Discovery of potent and selective reversible Bruton's tyrosine kinase inhibitors.

Bruton's tyrosine kinase (BTK) is a cytoplasmic, non-receptor tyrosine kinase member of the TEC family of tyrosine kinases. Pre-clinical and clinical data have shown that targeting BTK can be used for the treatment for B-cell disorders. Here we disclose the discovery of a novel imidazo[4,5-b]pyridine series of potent, selective reversible BTK inhibitors through a rational design approach. From a starting hit molecule 1, medicinal chemistry optimization led to the development of a lead compound 30, which exhibited 58 nM BTK inhibitory potency in human whole blood and high kinome selectivity. Additionally, the compound demonstrated favorable pharmacokinetics (PK), and showed potent dose-dependent efficacy in a rat CIA model.

Overcoming the unexpected functional inversion of a PqsR antagonist in Pseudomonas aeruginosa: an in vivo potent antivirulence agent targeting pqs quorum sensing.

The virulence regulator PqsR of Pseudomonas aeruginosa is considered as an attractive target for attenuating the bacterial pathogenicity without eliciting resistance. However, despite efforts and desires, no promising PqsR antagonist has been discovered thus far. Now, a surprising functionality change of a highly affine PqsR antagonist in P. aeruginosa is revealed, which is mediated by a bacterial signal molecule synthase and responsible for low cellular potency. Blockade of the susceptible position led to the discovery of the first antivirulence compound that is potent in vivo and targets PqsR, thus providing a proof of concept for this novel antivirulence therapy.

Reducing pressure with the goal of improving outcomes: a retrospective chart review of cushion evaluations and recommendations at one seating clinic.

PURPOSE: To describe the current seating recommendations made by a seating clinic for wheelchair users who presented with a Pressure Injury (PrI) or history of PrI. METHODS: Retrospective review of electronic medical records of 133 adults who used a wheelchair as their primary means of mobility who had a cushion evaluation during which interface pressure mapping data was documented. RESULTS: Clinicians adjusted 71% of participants' wheelchair cushions, including 49% who received a new cushion, and 37% of participants' wheelchairs. The most common adjustments besides receiving a new cushion were: addition of an underlay, adjusting the inflation of a cushion, and adjustments to the foot or back support of the wheelchair. Forty-five participants only received adjustments (i.e. no new cushion), while 23 participants only received education and feedback rather than equipment modifications. Those 23 participants had significantly lower Peak Pressure Index (PPI) than those who received equipment modifications (mean [95% CI] 76.7 [59.1, 94.3] versus 111.6 [102.1, 121.2] respectively, p = 0.001). The PPI was reduced by an average of 22.5 mmHg from the initial to final seating system amongst those who received modifications ([13.9-31.0], p<.001). CONCLUSIONS: The seating clinicians considered interface pressure mapping in their decision-making and effectively reduced interface pressures with their interventions. Cushion replacement is important when someone presents with a PrI. However, adjusting an existing wheelchair cushion and/or seating system provides important additions and alternatives to consider for reducing interface pressure. There is also a role for education about proper use of equipment, weight shifts, and alternate seating surfaces.IMPLICATIONS FOR REHABILITATIONA cushion evaluation may involve evaluating more than one cushion configuration and using pressure mapping to compare the best options.To address perceived wheelchair cushion issues, posture and positioning should be evaluated and adjusted as necessary, in addition to evaluating the cushion itself.Common positioning modifications include: modifying/adding cushion underlays or inflation and foot and back supports in response to clients' changing postural needs and wheelchair components coming out of optimal position due to wear and tear.Adjustments to the wheelchair and cushion aim to distribute body weight over a larger surface area, reduce pressure at high-risk locations, improve posture, and increase function. These adjustments should consider individual's specific needs and goals, while also being mindful of funding barriers.

Validation of PqsD as an anti-biofilm target in Pseudomonas aeruginosa by development of small-molecule inhibitors.

2-Heptyl-4-hydroxyquinoline (HHQ) and Pseudomonas quinolone signal (PQS) are involved in the regulation of virulence factor production and biofilm formation in Pseudomonas aeruginosa. PqsD is a key enzyme in the biosynthesis of these signal molecules. Using a ligand-based approach, we have identified the first class of PqsD inhibitors. Simplification and rigidization led to fragments with high ligand efficiencies. These small molecules repress HHQ and PQS production and biofilm formation in P. aeruginosa. This validates PqsD as a target for the development of anti-infectives.

Influence of DNA template choice on transcription and inhibition of Escherichia coli RNA polymerase.

In recent decades, quantitative transcription assays using bacterial RNA polymerase (RNAP) have been performed under widely diverse experimental conditions. We demonstrate that the template choice can influence the inhibitory potency of RNAP inhibitors. Furthermore, we illustrate that the sigma factor (σ(70)) surprisingly increases the transcription efficiency of templates with nonphysiological nonprokaryotic promoters. Our results might be a useful guideline in the early stages of using RNAP for drug discovery.

Discovery of Covalent Bruton's Tyrosine Kinase Inhibitors with Decreased CYP2C8 Inhibitory Activity.

Bruton's tyrosine kinase (BTK) is a member of the Tec kinase family that is expressed in cells of hematopoietic lineage. Evidence has shown that inhibition of BTK has clinical benefit for the treatment of a wide array of autoimmune and inflammatory diseases. Previously we reported the discovery of a novel nicotinamide selectivity pocket (SP) series of potent and selective covalent irreversible BTK inhibitors. The top molecule 1 of that series strongly inhibited CYP2C8 (IC50 =100 nM), which was attributed to the bridged linker group. However, our effort on the linker replacement turned out to be fruitless. With the study of the X-ray crystal structure of compound 1, we envisioned the opportunity of removal of this liability via transposition of the linker moiety in 1 from C6 to C5 position of the pyridine core. With this strategy, our optimization led to the discovery of a novel series, in which the top molecule 18 A displayed reduced CYP inhibitory activity and good potency. To further explore this new series, different warheads besides acrylamide, for example cyanamide, were also tested. However, this effort didn't lead to the discovery of molecules with better potency than 18 A. The loss of potency in those molecules could be related to the reduced reactivity of the warhead or reversible binding mode. Further profiling of 18 A disclosed that it had a strong hERG (human Ether-a-go-go Related Gene) inhibition, which could be related to the phenoxyphenyl group.

Load redistribution in variable position wheelchairs in people with spinal cord injury.

BACKGROUND/OBJECTIVE: Tilt and recline variable position seating systems are most commonly used for pressure relief to decrease potential for skin breakdown. This study provides quantitative information on the magnitudes of loading on the seat and back during phases of tilt, recline, and standing. The objective of this study was to show that the amount of force reduction at the seat would differ across these 3 methods within their respective clinical ranges. PARTICIPANTS: Six able-bodied (AB) subjects (2 men, 4 women) with a median age of 25 years, and 10 subjects (8 men, 2 women) with spinal cord injury (SCI) with a median age of 35.5 years. METHODS: Subjects sat on a power wheelchair with Tekscan pressure mats placed underneath a foam backrest and cushion. Data were collected at 5 positions for each method. Order of position and method tested were randomized. Linear regressions were used to calculate the relationships of normalized seat and backrest forces to seat and backrest angles for each chair configuration. RESULTS: Normalized seat loads had strong linear relationships with the angles of change in tilt, recline, and standing for both groups. Maximum decreases in seat load occurred at full standing and full recline in the SCI subjects and in full standing in the AB subjects. Loads linearly increased on the back during tilt and recline and linearly decreased during standing for both groups. CONCLUSIONS: Standing and recline offered similar seat load reductions at their respective terminal positions. Standing also reduced loading on the backrest. Recognizing that each method had clinical benefits and drawbacks, the results of this study indicate that tilt, recline, and standing systems should be considered as a means of weight shifting for wheelchair users.

Flexible Fragment Growing Boosts Potency of Quorum-Sensing Inhibitors against Pseudomonas aeruginosa Virulence.

Hit-to-lead optimization is a critical phase in drug discovery. Herein, we report on the fragment-based discovery and optimization of 2-aminopyridine derivatives as a novel lead-like structure for the treatment of the dangerous opportunistic pathogen Pseudomonas aeruginosa. We pursue an innovative treatment strategy by interfering with the Pseudomonas quinolone signal (PQS) quorum sensing (QS) system leading to an abolishment of bacterial pathogenicity. Our compounds act on the PQS receptor (PqsR), a key transcription factor controlling the expression of various pathogenicity determinants. In this target-driven approach, we made use of biophysical screening via surface plasmon resonance (SPR) followed by isothermal titration calorimetry (ITC)-enabled enthalpic efficiency (EE) evaluation. Hit optimization then involved growth vector identification and exploitation. Astonishingly, the latter was successfully achieved by introducing flexible linkers rather than rigid motifs leading to a boost in activity on the target receptor and anti-virulence potency.

Characterization of power wheelchair use in the home and community.

OBJECTIVE: To characterize the use of power wheelchairs and to determine if multiple measures of mobility and occupancy jointly provide a more comprehensive picture of wheelchair usage and daily activity in full-time power wheelchair users than daily distance alone. DESIGN: Prospective observational study. SETTING: Subjects' everyday mobility was measured in their homes and communities for 2 weeks, and prompted recall interviews were conducted by phone. PARTICIPANTS: A convenience sample (N=25) of nonambulatory, full-time power wheelchair users. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Wheelchair usage was logged electronically, and geolocation and interview data were used to isolate chair use to (1) in the home, (2) not in the home indoors, or (3) outdoors. Distance wheeled, time spent wheeling, number of bouts, time spent in the wheelchair, and the percentage of time in the wheelchair spent wheeling were measured to describe wheelchair use. RESULTS: The median wheelchair user spent 10.6 hours (range, 5.0-16.6h) in his/her wheelchair daily and wheeled 1.085 km (range, 0.238-10.585 km) over 58 minutes (range, 16-173 min) and 110 bouts (range, 36-282 bouts). Wheelchair use varied across subjects, within subjects from day to day, and between environments. Mobility bouts outdoors were longer and faster than those wheeled indoors. In a regression analysis, distance wheeled explained only 33% of the variation in the number of bouts and 75% in the time spent wheeling. CONCLUSIONS: Power wheelchair use varies widely both within and between users. Measuring distance, time, and number of bouts provides a clearer picture of mobility patterns than measuring distance alone, whereas occupancy helps to measure wheelchair function in daily activities.

Crocadepsins-Depsipeptides from the Myxobacterium Chondromyces crocatus Found by a Genome Mining Approach.

Analysis of the genome sequence of the myxobacterium Chondromyces crocatus Cm c5 revealed the presence of numerous cryptic megasynthetase gene clusters, one of which we here assign to two previously unknown chlorinated metabolites by a comparative gene inactivation and secondary metabolomics approach. Structure elucidation of these compounds revealed a unique cyclic depsipeptide skeleton featuring ß- and δ-amide bonds of aspartic acid and 3-methyl ornithine moieties, respectively. Insights into their biosynthesis were obtained by targeted gene inactivation and feeding experiments employing isotope-labeled precursors. The compounds were produced ubiquitously by the species Chondromyces crocatus and were found to inhibit the carbon storage regulator-RNA interaction.

Binding Mode Characterization and Early in Vivo Evaluation of Fragment-Like Thiols as Inhibitors of the Virulence Factor LasB from Pseudomonas aeruginosa.

The increasing emergence of antibiotic resistance necessitates the development of anti-infectives with novel modes of action. Targeting bacterial virulence is considered a promising approach to develop novel antibiotics with reduced selection pressure. The extracellular collagenase elastase (LasB) plays a pivotal role in the infection process of Pseudomonas aeruginosa and therefore represents an attractive antivirulence target. Mercaptoacetamide-based thiols have been reported to inhibit LasB as well as collagenases from clostridia and bacillus species. The present work provides an insight into the structure-activity relationship (SAR) of these fragment-like LasB inhibitors, demonstrating an inverse activity profile compared to similar inhibitors of clostridial collagenase H (ColH). An X-ray cocrystal structure is presented, revealing distinct binding of two compounds to the active site of LasB, which unexpectedly maintains an open conformation. We further demonstrate in vivo efficacy in a Galleria mellonella infection model and high selectivity of the LasB inhibitors toward human matrix metalloproteinases (MMPs).

Development of valid and reliable measures of postural stability.

BACKGROUND/OBJECTIVE: The development of simple postural stability tests that relate to performance of activities of daily living (ADL) and can be quickly performed in a clinical setting may assist clinicians in determining appropriate wheelchair configurations and postural supports in an efficient manner. The study's purpose was to validate 3 clinical measures of reach-functional reach (FR), reach area (RA), and bilateral reach (BR)-against the performance of ADL tasks. METHODS: Two groups of 20 subjects differing by time since spinal cord injury were tested. Three measures of reach-FR, RA, and BR-were recorded with and without permitting compensatory strategies. Subjects also attempted a series of ADL tasks. Group 2 subjects participated in test-retest reliability of the reach measures and to measure reach while using compensatory strategies. Correlation, ANOVA, and linear regression were used for analysis. RESULTS: Regression analysis showed that injury level was a significant predictor of success in performing ADL tasks (%ADL). Significant but not strong correlations were found between %ADL and all uncompensated reach measures. Within Group 2 subjects, compensated FR (r = 0.663) and RA (r = 0.647) were more related to the %ADL score than the uncompensated FR (r = 0.348) and RA (r = 0.305) measurements. BR had the strongest relationship with %ADL scores (P = 0.031) and was the only significant uncompensated reach measurement within the regression analyses. DISCUSSION AND CONCLUSION: While working with clients on seated stability and functional movement, clinicians should be encouraged to incorporate BR tasks because it has the strongest relationship to ADL performance. Researchers interested in studying postural control and stability during functional tasks should consider using uncompensated reach measures.

In-depth Profiling of MvfR-Regulated Small Molecules in Pseudomonas aeruginosa after Quorum Sensing Inhibitor Treatment.

Pseudomonas aeruginosa is a Gram-negative bacterium, which causes opportunistic infections in immuno-compromised individuals. Due to its multiple resistances toward antibiotics, the development of new drugs is required. Interfering with Quorum Sensing (QS), a cell-to-cell communication system, has shown to be highly efficient in reducing P. aeruginosa pathogenicity. One of its QS systems employs Pseudomonas Quinolone Signal (PQS) and 4-hydroxy-2-heptylquinoline (HHQ) as signal molecules. Both activate the transcriptional regulator MvfR (Multiple Virulence Factor Regulator), also called PqsR, driving the production of QS molecules as well as toxins and biofilm formation. The aim of this work was to elucidate the effects of QS inhibitors (QSIs), such as MvfR antagonists and PqsBC inhibitors, on the biosynthesis of the MvfR-regulated small molecules 2'-aminoacetophenone (2-AA), dihydroxyquinoline (DHQ), HHQ, PQS, and 4-hydroxy-2-heptylquinoline-N-oxide (HQNO). The employed synthetic MvfR antagonist fully inhibited pqs small molecule formation showing expected sigmoidal dose-response curves for 2-AA, HQNO, HHQ and PQS. Surprisingly, DHQ levels were enhanced at lower antagonist concentrations followed by a full suppression at higher QSI amounts. This particular bi-phasic profile hinted at the accumulation of a biosynthetic intermediate resulting in the observed overproduction of the shunt product DHQ. Additionally, investigations on PqsBC inhibitors showed a reduction of MvfR natural ligands, while increased 2-AA, DHQ and HQNO levels compared to the untreated cells were detected. Moreover, PqsBC inhibitors did not show any significant effect in PA14 pqsC mutant demonstrating their target selectivity. As 2-AA is important for antibacterial tolerance, the QSIs were evaluated in their capability to attenuate persistence. Indeed, persister cells were reduced along with 2-AA inhibition resulting from MvfR antagonism, but not from PqsBC inhibition. In conclusion, antagonizing MvfR using a dosage capable of fully suppressing this QS system will lead to a favorable therapeutic outcome as DHQ overproduction is avoided and bacterial persistence is reduced.

Dissecting the Multiple Roles of PqsE in Pseudomonas aeruginosa Virulence by Discovery of Small Tool Compounds.

Pseudomonas aeruginosa uses quorum sensing (QS) as a cell-to-cell communication system to orchestrate the expression of virulence determinants. The biosynthesis of the important Pseudomonas quinolone signal (PQS) requires the pqsABCDE operon. Here, PqsE acts as a pathway-specific thioesterase, but it also contributes to the regulation of bacterial virulence via an unknown mechanism. In this manuscript, we report the discovery of PqsE inhibitors as tool compounds to gain further insights into its different functions. Differential scanning fluorimetry (DSF) was used to screen a fragment library, and isothermal titration calorimetry (ITC) was employed as a secondary filter. As proven by X-ray crystallography, hit molecules bound to the active center inhibiting PqsE's thioesterase activity in cell-based and in vitro assays. Notably, the ligands did not affect the levels of the PqsE-regulated virulence factor pyocyanin. These findings indicate that the regulatory function of PqsE is not linked to its thioesterase activity and must be encoded outside of the active center. This study highlights the potential of fragment-based screening for the discovery of tool compounds. This approach provided novel insight into complex biological systems, which could not be obtained by knockout studies.

Discovery of the first small-molecule CsrA-RNA interaction inhibitors using biophysical screening technologies.

AIM: CsrA is a global post-transcriptional regulator protein affecting mRNA translation and/or stability. Widespread among bacteria, it is essential for their full virulence and thus represents a promising anti-infective drug target. Therefore, we aimed at the discovery of CsrA-RNA interaction inhibitors. Results & methodology: We followed two strategies: a screening of small molecules (A) and an RNA ligand-based approach (B). Using surface plasmon resonance-based binding and fluorescence polarization-based competition assays, (A) yielded seven small-molecule inhibitors, among them MM14 (IC50 of 4 µM). (B) resulted in RNA-based inhibitor GGARNA (IC50 of 113 µM). CONCLUSION: The first small-molecule inhibitors of the CsrA-RNA interaction were discovered exhibiting micromolar affinities. These hits represent tools to investigate the effects of CsrA-RNA interaction inhibition on bacterial virulence.

Effect of seat inclination on seated pressures of individuals with spinal cord injury.

BACKGROUND AND PURPOSE: Manual wheelchair configurations commonly include "squeezing" the wheelchair frame to improve balance for users with spinal cord injuries. This squeezing is achieved by lowering the rear portion of the seat relative to the front of the seat while maintaining the same back angle. The study's purpose was to examine the effect of increasing posterior seat inclination on buttock interface pressures. SUBJECTS: Nine male and 5 female subjects (mean age=37 years, SD=11.2, range=19-55) with complete thoracic or lumbar spinal cord injury were tested. METHODS: Subjects sat on a pressure mat placed over a foam cushion. Pressure readings were taken at seat angles reflecting seat height decreases of 0, 5.1, 7.6, and 10.2 cm (0, 2, 3, and 4 in) of the rear of the seat relative to the front of the seat. An analysis of variance and a Duncan multiple range test were used for data analysis. RESULTS: No meaningful differences were found in measurements of interface pressure (dispersion index, contact area, and seat pressure index), total force on seat, or peak pressure index with posterior seat inclination. DISCUSSION AND CONCLUSION: The data indicate no meaningful evidence that squeezing a wheelchair frame increases seat interface pressures.

Optimization of anti-virulence PqsR antagonists regarding aqueous solubility and biological properties resulting in new insights in structure-activity relationships.

Increasing antibiotic resistance urgently requires novel therapeutic options to combat bacterial infections. The anti-virulence therapy selectively intervening with pathogenicity without affecting bacterial viability is such a strategy to overcome resistance. We consider the virulence regulator PqsR as an attractive target in the human pathogen Pseudomonas aeruginosa, and recently discovered the first PqsR antagonists, which, however, suffered from poor aqueous solubility. In this work, the antagonists were structurally modified to become more soluble, and their structure-activity as well as structure-property relationships were studied. A novel promising compound with improved solubility and enhanced anti-virulence activity was discovered (IC50: 3.8 µM, pyocyanin). Our findings emphasize the crucial role of substituents at the 3-position and the carbonyl group at the 4-position for ligand-receptor interactions, and illuminate the way for further optimization of PqsR antagonists as anti-virulence agents.

Antibiotic-free nanotherapeutics: ultra-small, mucus-penetrating solid lipid nanoparticles enhance the pulmonary delivery and anti-virulence efficacy of novel quorum sensing inhibitors.

Cystic fibrosis (CF) is a genetic disease mainly manifested in the respiratory tract. Pseudomonas aeruginosa (P. aeruginosa) is the most common pathogen identified in cultures of the CF airways, however, its eradication with antibiotics remains challenging as it grows in biofilms that counterwork human immune response and dramatically decrease susceptibility to antibiotics. P. aeruginosa regulates pathogenicity via a cell-to-cell communication system known as quorum sensing (QS) involving the virulence factor (pyocyanin), thus representing an attractive target for coping with bacterial pathogenicity. The first in vivo potent QS inhibitor (QSI) was recently developed. Nevertheless, its lipophilic nature might hamper its penetration of non-cellular barriers such as mucus and bacterial biofilms, which limits its biomedical application. Successful anti-infective inhalation therapy necessitates proper design of a biodegradable nanocarrier allowing: 1) high loading and prolonged release, 2) mucus penetration, 3) effective pulmonary delivery, and 4) maintenance of the anti-virulence activity of the QSI. In this context, various pharmaceutical lipids were used to prepare ultra-small solid lipid nanoparticles (us-SLNs) by hot melt homogenization. Plain and QSI-loaded SLNs were characterized in terms of colloidal properties, drug loading, in vitro release and acute toxicity on Calu-3 cells. Mucus penetration was studied using a newly-developed confocal microscopy technique based on 3D-time-lapse imaging. For pulmonary application, nebulization efficiency of SLNs and lung deposition using next generation impactor (NGI) were performed. The anti-virulence efficacy was investigated by pyocyanin formation in P. aeruginosa cultures. Ultra-small SLNs (<100nm diameter) provided high encapsulation efficiency (68-95%) according to SLN composition, high burst in phosphate buffer saline compared to prolonged release of the payload over >8h in simulated lung fluid with minor burst. All types and concentrations of plain and QSI-loaded SLNs maintained the viability of Calu-3 cells. 3D time-lapse confocal imaging proved the ability of SLNs to penetrate into artificial sputum model. SLNs were efficiently nebulized; NGI experiments revealed their deposition in the bronchial region. Overall, nanoencapsulated QSI showed up to sevenfold superior anti-virulence activity to the free compound. Most interestingly, the plain SLNs exhibited anti-virulence properties themselves, which was shown to be related to anti-virulence effects of the emulsifiers used. These startling findings represent a new perspective of ultimate significance in the area of nano-based delivery of novel anti-infectives.

Development and validation of a UHPLC-MS/MS procedure for quantification of the Pseudomonas Quinolone Signal in bacterial culture after acetylation for characterization of new quorum sensing inhibitors.

The appearance of antibiotic resistance requires novel therapeutic strategies. One approach is to selectively attenuate bacterial pathogenicity by interfering with bacterial cell-to-cell communication known as quorum sensing. The PQS quorum sensing system of Pseudomonas aeruginosa employs as signal molecule the Pseudomonas Quinolone Signal (PQS; 2-heptyl-3-hydroxy-4-(1H)-quinolone), a key contributor to virulence and biofilm formation. Thus, interference with PQS production is considered as promising approach for the development of novel anti-infectives. Therefore, in this study, we developed and validated an ultra-high performance liquid chromatographic-tandem mass spectrometric approach for reliable quantification of PQS in P. aeruginosa cultures for activity determination of new quorum sensing inhibitors. The poor chromatographic properties of PQS reported by others could be overcome by fast microwave-assisted acetylation. The validation procedure including matrix effects, recovery, process efficiency, selectivity, carry-over, accuracy and precision, stability of the processed sample, and limit of quantification demonstrated that the method fulfilled all requirements of common validation guidelines. Its applicability was successfully proven in routine testing. In addition, two-point calibration was shown to be applicable for fast and reliable PQS quantification saving time and resources. In summary, the described method provides a powerful tool for the discovery of new quorum sensing inhibitors as potential anti-infectives and illustrated the usefulness of chemical derivatization, acetylation, in liquid chromatography-mass spectrometry analysis.