Bioinformatics approaches for the analysis of lipidomics data

The potential impact of lipid research has been increasingly realised both in disease treatment and prevention. Recent advances in soft ionization mass spectrometry (MS) such as electrospray ionization (ESI) have permitted parallel monitoring of several hundreds of lipids in a single experiment and thus facilitated lipidomics level studies. These advances, however, pose a greater challenge for bioinformaticians to handle massive amounts of information-rich MS data from modern analytical instruments in order to understand complex functions of lipids. The main aims of this thesis were to 1) develop bioinformatics approaches for lipid identification based on ultra performance liquid chromatography coupled to mass spectrometry (UPLC/MS) data, 2) predict the functional annotations for unidentified lipids, 3) understand the omics data in the context of pathways and 4) apply existing chemometric methods for exploratory data analysis as well as biomarker discovery. A bioinformatics strategy for the construction of lipid database for major classes of lipids is presented using simplified molecular input line entry system (SMILES) approach. The database was annotated with relevant information such as lipid names including short names, SMILES information, scores, molecular weight, monoisotopic mass, and isotope distribution. The database was tailored for UPLC/MS experiments by incorporating the information such as retention time range, adduct information and main fragments to screen for the potential lipids. This database information facilitated building experimental tandem mass spectrometry libraries for different biological tissues. Non-targeted metabolomics screening is often get plagued by the presence of unknown peaks and thus present an additional challenge for data interpretation. Multiple supervised classification methods were employed and compared for the functional prediction of class labels for unidentified lipids to facilitate exploratory analysis further as well as ease the identification process. As lipidomics goes beyond complete characterization of lipids, new strategies were developed to understand lipids in the context of pathways and thereby providing insights for the phenotype characterization. Chemometric methods such as principal component analysis (PCA) and partial least squares and discriminant analysis (PLS/DA) were utilised for exploratory analysis as well as biomarker discovery in the context of different disease phenotypes

Identification of triterpenoids in Brachylaena discolor DC, a plant with antidiabetic activity.

Masters Degree. University of KwaZulu-Natal, Pietermaritzburg.In recent times, metabolic diseases such as diabetes and hypertension have become a huge burden in populations around the world. Among these diseases, the number of diabetes mellitus cases have been increasing at an alarming rate. Problems such as poor access to medication and side effects of existing drugs are encountered and there is still a need for more therapeutic alternatives. Plants have been used extensively for the treatment of various diseases, among them diabetes. Brachylaena discolor DC is a South African medicinal plant that is used by people for the treatment of diabetes and it has been reported previously to have antidiabetic activity with little toxicity. This study aimed to investigate the triterpenoid content of B. discolor and further evaluate the plant for in vitro antidiabetic activity by inhibition of the enzyme α-glucosidase, a therapeutic target for treatment of diabetes. In literature, it has been reported that some triterpenoids are good inhibitors of α-glucosidase. Chromatographic techniques, including argentation chromatography on silica gel impregnated with silver nitrate, were used in the isolation of compounds. The compounds were identified by 1D and 2D NMR (i.e. 1H, 13C, COSY, DEPT, HSQC and HMBC), IR, GC-MS, and HR-MS. From the dichloromethane extract of B. discolor leaves, eight triterpenoids were isolated, α-amyrin acetate, β-amyrin acetate, ψ-taraxasterol acetate, taraxasterol acetate, lupeol acetate, α-amyrin palmitate, β-amyrin palmitate, and lupeol palmitate. All the compounds are reported for the first time from the plant, except lupeol acetate, which was previously identified in the plant. Hydrolysis of α-amyrin acetate, β-amyrin acetate, ψ-taraxasterol acetate, taraxasterol acetate, and lupeol acetate afforded the alcohol triterpenoids α-amyrin, β-amyrin, ψ-taraxasterol, taraxasterol, and lupeol. The DCM-MeOH (1:1) leaf extract of B. discolor was investigated in vitro against α-glucosidase and the extract was to inhibit the enzyme significantly with an IC50 value of 95.95 μg/mL when compared with the standard inhibitor acarbose with an IC50 value of 1149.07 μg/mL. The individual pure compounds were not assayed due to poor solubility in the assay medium. Molecular docking studies revealed that the triterpenoids, α-amyrin, β-amyrin, ψ-taraxasterol, taraxasterol, and lupeol all have binding affinity for the α-glucosidase active site, where α-amyrin and β-amyrin showed more pronounced results with binding energies of -8.90 and -8.00 kcal/mol, respectively. In the study the reported antidiabetic activity of B. discolor was corroborated

Identification and Targeting of Novel Mechanisms for Treatment of Castration Resistant Prostate Cancer (CRPC)

Prostate cancer (PCa) is a significant contributor to cancer-related deaths among men globally, with varying prevalence across regions, particularly higher rates in North America, Europe, and Australia. The disease's incidence escalates with age, emphasizing its relevance in aging populations. PCa cells are fueled by androgens through the Androgen Receptor (AR), which activates genes crucial for tumor growth and survival. Thus, reducing circulating androgen levels or blocking AR remains primary in PCa treatment. However, in advanced stages like Castration-Resistant Prostate Cancer (CRPC), tumor cells develop mechanisms to thrive independently of androgens. These mechanisms involve the production of androgens within the tumor itself, highlighting androgen biosynthesis as a key target for CRPC treatment. Androgens, primarily produced in the testes, ovaries, and adrenal glands, regulate male sexual traits and behaviors. The adrenal cortex's Zona Reticularis (ZR) produces precursor hormones like dehydroepiandrosterone (DHEA) and its sulfate, which serve as substrates for androgen production. Cytochrome P450c17 (CYP17A1) plays a pivotal role in adrenal androgen production, catalyzing essential reactions in steroidogenesis. Given its significance, inhibiting CYP17A1 has garnered interest in PCa treatment. Abiraterone, a CYP17A1 inhibitor, albeit effective, lacks specificity and affects other CYP enzymes. Recent discoveries, like mutations abolishing CYP17A1's specific lyase activity, offer hope for developing selective inhibitors. Efforts focus on designing molecules targeting CYP17A1 without resembling androgens to avoid inadvertently stimulating androgen receptor signaling. Small molecule inhibitors are being screened, including those computationally designed to mimic native substrates. Additionally, exploration into endocrine disruptors' role in inhibiting CYP17A1 activity provides insights into environmental factors influencing cancer risk. Understanding the regulation of CYP17A1 activity through phosphorylation pathways offers another avenue. Kinases such as p38α and ROCK1, along with phosphatases like PP2A, play roles in modulating CYP17A1 activity. PLK1 inhibition demonstrated decreased CYP17A1 lyase activity, suggesting its potential as a target. Transcriptomic analysis identified differentially expressed kinases and phosphatases in PCa and polycystic ovary syndrome (PCOS), shedding light on potential therapeutic targets. Insights into CYP17A1 regulation also have implications beyond PCa, extending to hyperandrogenic disorders like PCOS and premature adrenarche. Developing drugs to lower androgen levels without affecting cortisol can mitigate side effects seen with current therapies, offering broader applications in reproductive health. In conclusion, targeting CYP17A1 holds promise in PCa treatment, especially in CRPC. Advances in understanding its regulation and identifying specific inhibitors offer hope for more effective therapies. Moreover, insights into CYP17A1's role in hyperandrogenic disorders present opportunities for broader therapeutic interventions in reproductive health

SIGMA 1 RECEPTOR (S1R) MODULATORS AS A THERAPEUTIC STRATEGY FOR PROMOTING NEUROPLASTICITY. DESIGN AND SYNTHESIS OF NOVEL MONO- AND BI-VALENT LIGANDS FOR SRS

During my three-years-project I focused on the development of novel mono- and bi-valent Sigma1 Receptor (S1R) modulators to address two main objectives: (i) the obtainment of multitarget-directed ligands (MTDLs) endowed with therapeutic potential for the treatment of neurodegenerative diseases; (ii) the preparation of a series of bivalent compounds to be used for the study of S1R oligomerization process. These two major topics are briefly discussed hereafter. (i) Neurodegeneration is a key event in many challenging disorders (e.g. Alzheimers diseases, Parkinsons disease, multiple sclerosis). Such pathologies involve the alteration of several molecular pathways, making the multi-target paradigm a promising strategy for new effective therapies. Among the numerous molecular targets that have been correlated with neurodegenerative disorders, S1R has gained great attention from the scientific community, and S1R agonists are considered viable pharmacological tools for their neuroprotective activity. Hence, we reasoned that coupling S1R agonism with modulation of other selected targets might afford new molecular entities more effective in counteracting neuropathies. The additional targets of our MTDLs include N-Methyl-D-Aspartate (NMDA) receptor, which plays a relevant role in synaptic plasticity, and acetylcholinesterase (AChE), which regulates acetylcholine levels in central nervous system. A structurally focused compound library was prepared through a divergent synthesis. The so-obtained compounds were tested for a preliminary biological evaluation, evaluating their affinity and selectivity towards S1R and NMDA receptor, the AChE inhibition and their antioxidant properties, since oxidative stress is considered a hallmark of neurodegeneration. A number of promising compounds, endowed with effective multitarget profile, was identified. These results will pave the way for further biological investigation and structure optimization in order to achieve viable tools for the treatment of neurodegenerative diseases. (ii) In the last decade numerous studies have supported the hypothesis that S1R can exist in multiple oligomeric forms. In detail, agonists seem to stabilize S1R monomers and dimers that act as chaperones, whereas antagonists bind to higher oligomer complexes, maintaining them in repository forms. Moreover, the recently disclosed crystal of S1R was obtained as a trimer. Nevertheless, the mechanism of generation, as well as the precise biological function of S1R oligomers, are still unknown. Accordingly, a series of homo- and hetero-bivalent S1R ligands was designed and synthetized to investigate S1R oligomerization process. Since S1R agonists are known to exert neuroprotective effects, and S1R can form homo-dimeric structures upon interaction with agonists, we reasoned that promoting dimerization through bivalent agonists might enhance ligands activity. The designed bivalent compounds consist in two units of (R)-RC-33 (a potent and selective S1R agonist developed by our group) joined by a linker. Different lengths, polarities and spatial constraints were explored for the linker. The key precursor of the synthesis is (R)-RC-33A, an aminic derivative of RC-33. For the obtainment of enantiopure (R)-RC-33A, three different synthetic approaches have been explored, resulting in the identification of an efficient pathway to access (R)-RC-33 derivatives with high yield and chiral purity. Once the designed ligands were obtained in sufficient amount and purity, they were tested in binding assays to assess their S1R affinity. Moreover, computational studies were performed on both mono- and bi-valent S1R modulators. In detail, docking into the crystals binding pocket served as basis for the development of a 3D-QSAR model and for the rationalization of experimental results. Molecular dynamics studies are ongoing, and future functional assays will contribute to shed light on the S1R oligomeric states

Phenotypic Screening of Chemical Libraries Enriched by Molecular Docking to Multiple Targets Selected from Glioblastoma Genomic Data

Like most solid tumors, glioblastoma multiforme (GBM) harbors multiple overexpressed and mutated genes that affect several signaling pathways. Suppressing tumor growth of solid tumors like GBM without toxicity may be achieved by small molecules that selectively modulate a collection of targets across different signaling pathways, also known as selective polypharmacology. Phenotypic screening can be an effective method to uncover such compounds, but the lack of approaches to create focused libraries tailored to tumor targets has limited its impact. Here, we create rational libraries for phenotypic screening by structure-based molecular docking chemical libraries to GBM-specific targets identified using the tumor’s RNA sequence and mutation data along with cellular protein–protein interaction data. Screening this enriched library of 47 candidates led to several active compounds, including 1 (IPR-2025), which (i) inhibited cell viability of low-passage patient-derived GBM spheroids with single-digit micromolar IC50 values that are substantially better than standard-of-care temozolomide, (ii) blocked tube-formation of endothelial cells in Matrigel with submicromolar IC50 values, and (iii) had no effect on primary hematopoietic CD34+ progenitor spheroids or astrocyte cell viability. RNA sequencing provided the potential mechanism of action for 1, and mass spectrometry-based thermal proteome profiling confirmed that the compound engages multiple targets. The ability of 1 to inhibit GBM phenotypes without affecting normal cell viability suggests that our screening approach may hold promise for generating lead compounds with selective polypharmacology for the development of treatments of incurable diseases like GBM

Versuch des Ausbruchs aus dem Flachland: Erzeugung und biologische Profilierung verbrückt bicyclischer Verbindungen

The synthesis of new chemical entities as potential drug candidates is a promising and challenging aspect in drug discovery. Investigations on success rates showed higher values for three-dimensional molecules than for planar ones and recommended an ôescape from flatlandö strategy for the chemical synthesis of compound collections. Following these suggestions, a synthetic route was devised for the generation of bridged bicyclic compounds as an example for molecules possessing a permanent three-dimensional structure. This route involved [4+2] cycloaddition reactions between a collection of cyclic dienes and a collection of dienophiles, thereby combining privileged core structures provided by the reaction partners. A considerable fraction of the novel molecules showed biological activity in a general assay based on interference with the proliferation of mammalian cells. Two profiling methods, cellular impedance measurement and high-content cellular image analysis, were used to obtain suggestions for the types of bioactivities exerted by the positive compounds. Although the results of these experiments showed considerable variation, it was in two instances possible to identify cellular processes with which the active compounds interfere. In one case, the inhibition of tubulin polymerization was observed, in the other case, interference with proteasome function could be confirmed. The bridged bicyclic compounds synthesized and examined in the course of this thesis are just one example for structures that possess a permanent three-dimensionality. The results obtained encourage further exploration of this region of the chemical space by generation of larger and more structurally diverse collections of bridged bicyclic compounds and related molecules. Furthermore, other synthetic pathways, leading to different types of three-dimensional compounds, may be developed and explored.Die Synthese „neuer chemischer Einheiten“ zur Gewinnung von Wirkstoff-Kandidaten ist ein vielversprechender und herausfordernder Aspekt der Wirkstoffforschung. Untersuchungen über Erfolgsraten ergaben höhere Werte für dreidimensionale Moleküle als für planare und legten eine "Ausstieg-aus-dem-Flachland"-Strategie für die Synthese von Molekülbibliotheken nahe. In Verfolgung dieses Vorschlags wurde eine Syntheseroute für die Erzeugung verbrückt bizyklischer Verbindungen als ein Beispiel für Moleküle mit permanent dreidimensionaler Struktur entwickelt. Diese Route beinhaltete [4+2]-Zykloadditionsreaktionen zwischen Kollektionen von zyklischen Dienen und von Dienophilen, wobei durch die Reaktionspartner privilegierte Kernstrukturen miteinander kombiniert wurden. Ein signifikanter Teil der neuen Moleküle zeigte in einem globalen Test, der die Beeinträchtigung der Proliferation von Säugerzellen maß, biologische Aktivität. Zwei Profilierungs-Methoden, Verfolgung der zellulären Impedanz und extensive zelluläre Bildanalyse, wurden eingesetzt, um Hinweise auf die Arten der Bioaktivitäten der positiven Verbindungen zu erhalten. Obwohl die Ergebnisse dieser Experimente beträchtliche Streuungen zeigten, war es in zwei Fällen möglich, die zellulären Prozesse zu identifizieren, mit denen die aktiven Verbindungen interferierten. In einem Fall wurde die Inhibition der Tubulin-Polymerisation beobachtet, im anderen Fall konnte eine Störung der Proteasom-Funktion bestätigt werden. Die synthetisierten Verbindungen sind nur ein Beispiel für Moleküle, die eine permanente Dreidimensionalität besitzen. Die erzielten Ergebnisse sprechen für eine weitere Untersuchung dieser Region des „chemischen Raums“ durch Erzeugung größerer und strukturell vielfältigerer Sammlungen derartiger Verbindungen und verwandter Moleküle. Ferner könnten andere Synthesewege, die zu alternativen Typen dreidimensionaler Verbindungen führen, entworfen und auf bioaktive Moleküle untersucht werden

Antimicrobial and Resistance-Modifying Activities of LY2183240 Regioisomers

The rapid development of antimicrobial resistance over the past three decades represents a critical public health threat and urgent challenge. In this context, in order to establish lead compounds or identify novel modes of action one approach is to re-investigate drugs that affect eukaryotic processes for antimicrobial activity, particularly if their drug targets have homologies with bacterial proteins. In this study, regioisomers of LY2183240, a potent inhibitor of anandamide transport and fatty acid amide hydrolase were selected after a small screening study. The 2,5-LY2183240 regioisomer was shown to possess potent antimicrobial activity selective towards certain Gram-positive bacteria, which included Staphylococcus aureus and Bacillus subtilis, but not Enterococcus faecalis or Streptococcus pneumoniae. Conversely, the 1,5-LY2183240-regioisomer had no anti-bacterial activity strongly implicating the position of the carbamoyl on the tetrazole in the structure-activity relationship of the molecule. Investigation of the mechanism of antimicrobial activity suggested that while 2,5- LY2183240 had bacteriostatic activity this was probably not due to inhibition of protein or teichoic acid synthesis. Nevertheless, this activity may be related to the inhibition of bacterial fatty acid synthesis. Supporting this hypothesis, addition of exogenous fatty acids within Tween 80 was able to compromise 2-5-LY2183240 anti-staphylococcal activity. Based on the spectrum of activity and known redundancies at each of the steps in the fatty acid synthesis pathway, the most likely target was deduced to be FabI. However, characterization of a 2,5-LY2183240- resistant mutant revealed no alteration to the deduced amino acid sequence of FabI or relevant changes to FabI protein expression, an observation confirmed by analysis of the fabI promoter region and western blot studies. Since resistance to 2,5- LY2183240 could be mediated through non-target related factors such as drug efflux or drug entry into the cell, FabI could not be ruled out as a potential target. In addition, whole cell protein profiling revealed the 2,5-LY2183240-mutant to have differences in the expressions of several proteins, suggesting resistance may occur through a more global effect. Due to the promiscuous nature of LY2183240, the possibility of having multiple targets in S. aureus that collectively exhibit a bacteriostatic effect cannot be ruled out. This study also showed that the LY2183240 regioisomers are specific inhibitors of class C β-lactamases with optimum inhibitory activity dependent upon the position of the carbonyl on the tetrazole heterocyclic group (Ki values of 1.8 and 2.45 μM for 1,5- and 2,5-LY2183240, respectively). Molecular modelling suggested that LY2183240 regioisomers bind within the catalytic site of class C β-lactamases, interacting with some of the same residues, including Tyr150, Lys315 and Thr316, used by the substrate nitrocefin and β-lactamase inhibitors. The results substantiate the competitive inhibition model determined by enzyme kinetic studies. Furthermore, mass spectrometry data revealed non-covalent interactions between AmpC β- lactamase and LY2183240 regioisomers, which together with the non-reversible inhibitory activity, may indicate that both regioisomers present a high affinity towards this cephalosporinase and dissociate very slowly from the enzyme. LY2183240 may prove useful as a chemical scaffold for the development of novel and highly selective inhibitors of class C β-lactamases

Phytochemical Investigation and Bioactivity Assessment of Medicinal Plant from Northern Nigeria

Boswellia dalzielii (Burseraceae) has ethnopharmacological importance and is claimed to have anti-infection and immunomodulatory effects. In the Northern part of Nigeria, a region with a tropical dry climate, an aqueous infusion of this plant is used in the treatment of infections and tumours. The traditional formulation method was mimicked under laboratory conditions, and the effect of temperature and the impact of endophytic microbes present in aqueous infusion of B. dalzielii was also investigated. Activity-guided fractionation against Staphylococcus aureus and its methicillin-resistant strain resulted in the identification of two antibacterial compounds namely gallic acid and pyrogallol. The Minimum Inhibitory Concentration for pyrogallol and gallic acid against S. aureus growth are 508 and 753 μM, while against MRSA growth are 254 and 2032 μM, respectively. A growth Inhibition assay showed the activity of gallic acid as bacteriostatic, and pyrogallol as bacteriocidal against tested microorganisms. Interestingly, the bacteriocidal compound was found to arise by conversion of gallic acid by the endophyte Enterobacter cloacae. In addition, Pantoea spp was also isolated from the bark of B. Dalzielii. The sequences of both E. cloacae and Pantoea spp are deposited in the GenBank nucleotide database under the accession number MH764584 and MH764583, respectively. Similarly, activity-guided fractionation of B. Dalzielii bark against breast cancer cell line (MCF7) using MTT cytotoxicity assay resulted in the identification of a cytotoxic compound, catechol, and the half maximal effective concentration (EC50) observed was 86μM. The growth inhibition effect of catechol was observed to be time- and concentration- dependent. Endophytic Klebsiella pneumonia species (strain A and B) were shown to be responsible for bioconversion of protocatechuic acid to catechol. In addition, Pantoea agglomerans was also isolated from the bark of B. dalzielii. The sequences of Klebsiella pneumonia A, Klebsiella pneumonia B and Pantoea agglomerans are deposited in the GenBank nucleotide database under the accession number MH762022, MH762023 and MH762024, respectively. All isolated compounds were identified using HPLC, TLC, NMR, FTIR and HRMS.TETFUN

Screening for aerobic endospore-forming bacteria as biocontrol agents for powdery mildew disease of cucurbits.

Master of Science in Microbiology. University of KwaZulu-Natal, Pietermaritzburg 2015.Powdery mildew of cucurbits costs the South African cucurbit-growing industry millions of Rands per year in reduced yields and compromised fruit quality. Amongst the many bacterial and fungal antagonists of cucurbit powdery mildew, certain aerobic endospore-forming bacteria (AEFB) species show promise as biocontrol agents of this disease. When embarking upon biocontrol agent selection, multifaceted screening strategies are crucial. A study was undertaken with the aim of isolating AEFB from the cucurbit phylloplane for evaluation as potential antagonists of cucurbit powdery mildew using various screening approaches. Three hundred and nine AEFB isolates were isolated from cucurbit leaf material sourced from eight locations in the greater Msunduzi, KZN region. Dual-culture antifungal bioassays were performed using surrogate phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani in place of the obligately biotrophic Podosphaera spp.. Two PCR-based genotyping methods were used to differentiate and group 55 antifungal AEFB isolates: internal-transcribed spacer region (ITS) PCR and randomly amplified polymorphic DNA (RAPD) PCR. The RAPD-PCR distinguished greater levels of genetic polymorphisms amongst isolates than did the ITS-PCR, revealing 14 different profiles as opposed to the three obtained from ITS-PCR; with 42% of isolates associated with a single RAPD-PCR banding profile. Phylogenetic relationships between representatives of each of the RAPD-PCR fingerprint groupings were determined by sequence analysis of 16S rRNA and gyrase subunit A (gyrA) gene fragments. In each instance, several distinct clusters were discernable, though gyrA sequences displayed higher levels of strain-level sequence heterogeneity. Comparisons of both gene sequence types with reference strains from the GenBank database revealed similarities to several known plant-associated strains of AEFB, including B. amyloliquefaciens subsp. plantarum and B. subtilis. Matrix-assisted laser deionisation-desorption time-of-flight mass spectrometry (MALDI-TOF-MS) based identification of selected AEFB was evaluated by comparing spectral data from AEFB isolates with reference strains in a Bruker BDAL Biotyper database. Only three out of the 14 isolates evaluated were identified to species level with acceptable confidence levels. This poor taxonomic resolution was ascribed to a paucity of applicable reference strains in the BDAL library. Nevertheless, mass spectra profiles of each isolate allowed for the clustering of related isolates to be achieved when dendograms were created. Antifungal compounds were extracted from 14 isolates using an acid-precipitation and methanol extraction protocol. Detection and identification of lipopeptide compounds in these extracts was assessed using thin-layer chromatography (TLC) and MALDI-TOF-MS. PCR-based screening for lipopeptide production potential using selected lipopeptide gene markers (viz. surfactin, iturin, bacillomycin, and fengycin) was also evaluated for the selected 14 isolates. These isolates were found to produce multiple lipopeptide compounds; including homologues of surfactin, iturin, and fengycin. However, disparities that emerged between PCR, TLC, and MALDI-TOF-MS data suggest that some PCR primers, the ituD marker in particular, showed limited specificity amongst the AEFB strains screened. Based on the overall findings, nine isolates proceeded to in vivo screening against Podosphaera spp. using an agarised detached cotyledon assay and a biocontrol pot trial. Isolates achieving the most effective antagonism of Podosphaera spp. differed in each respective assay. Isolate cce175 provided the highest antagonism in the biocontrol pot trial, and isolate sqo279 provided the best results in the detached cotyledon assay. The impacts of inoculum preparation were assessed using isolate cce175 in a biocontrol pot trial. Treatments varied in cell growth phase and assessed cell-free supernatant, whole broth, and cell-only fractions on biocontrol efficacy compared to a Tebuconazole (430 g/l) fungicide control. None of the treatments were found to impact disease at a statistically significant level. The merits and limitations of the various screening approaches used, and issues surrounding the isolation and assessment of biocontrol efficacy in plant-associated AEFB, are discussed

Biological activities and ADMET-related properties of novel set of cinnamanilides

A series of nineteen novel ring-substituted N-arylcinnamanilides was synthesized and characterized. All investigated compounds were tested against Staphylococcus aureus as the reference strain, two clinical isolates of methicillin-resistant S. aureus (MRSA), and Mycobacterium tuberculosis. (2E)-N-[3-Fluoro-4-(trifluoromethyl)phenyl]-3-phenylprop-2-enamide showed even better activity (minimum inhibitory concentration (MIC) 25.9 and 12.9 M) againstMRSAisolates than the commonly used ampicillin (MIC 45.8 M). The screening of the cell viability was performed using THP1-Blue NF- B cells and, except for (2E)-N-(4-bromo-3-chlorophenyl)-3-phenylprop-2-enamide (IC50 6.5 M), none of the discussed compounds showed any significant cytotoxic e ect up to 20 M. Moreover, all compounds were tested for their anti-inflammatory potential; several compounds attenuated the lipopolysaccharide-induced NF- B activation and were more potent than the parental cinnamic acid. The lipophilicity values were specified experimentally as well. In addition, in silico approximation of the lipophilicity values was performed employing a set of free/commercial clogP estimators, corrected afterwards by the corresponding pKa calculated at physiological pH and subsequently cross-compared with the experimental parameters. The similarity-driven property space evaluation of structural analogs was carried out using the principal component analysis, Tanimoto metrics, and Kohonen mapping