| 1. |
- Alalam, Hanna, et al.
(författare)
-
A Genetic Trap in Yeast for Inhibitors of SARS-CoV-2 Main Protease
- 2021
-
Ingår i: MSYSTEMS. - 2379-5077. ; 6:6
-
Tidskriftsartikel (refereegranskat)abstract
- The ongoing COVID-19 pandemic urges searches for antiviral agents that can block infection or ameliorate its symptoms. Using dissimilar search strategies for new antivirals will improve our overall chances of finding effective treatments. Here, we have established an experimental platform for screening of small molecule inhibitors of the SARS-CoV-2 main protease in Saccharomyces cerevisiae cells, genetically engineered to enhance cellular uptake of small molecules in the environment. The system consists of a fusion of the Escherichia coli toxin MazF and its antitoxin MazE, with insertion of a protease cleavage site in the linker peptide connecting the MazE and MazF moieties. Expression of the viral protease confers cleavage of the MazEF fusion, releasing the MazF toxin from its antitoxin, resulting in growth inhibition. In the presence of a small molecule inhibiting the protease, cleavage is blocked and the MazF toxin remains inhibited, promoting growth. The system thus allows positive selection for inhibitors. The engineered yeast strain is tagged with a fluorescent marker protein, allowing precise monitoring of its growth in the presence or absence of inhibitor. We detect an established main protease inhibitor by a robust growth increase, discernible down to 1 mM. The system is suitable for robotized large-scale screens. It allows in vivo evaluation of drug candidates and is rapidly adaptable for new variants of the protease with deviant site specificities. IMPORTANCE The COVID-19 pandemic may continue for several years before vaccination campaigns can put an end to it globally. Thus, the need for discovery of new antiviral drug candidates will remain. We have engineered a system in yeast cells for the detection of small molecule inhibitors of one attractive drug target of SARS-CoV-2, its main protease, which is required for viral replication. The ability to detect inhibitors in live cells brings the advantage that only compounds capable of entering the cell and remain stable there will score in the system. Moreover, because of its design in yeast cells, the system is rapidly adaptable for tuning the detection level and eventual modification of the protease cleavage site in the case of future mutant variants of the SARSCoV-2 main protease or even for other proteases.
|
|
| 2. |
- Bourgard, Catarina, 1985, et al.
(författare)
-
Development of Dicationic Bisguanidine-Arylfuran Derivatives as Potent Agents against Gram-Negative Bacteria.
- 2022
-
Ingår i: Antibiotics. - : MDPI AG. - 2079-6382. ; 11:8
-
Tidskriftsartikel (refereegranskat)abstract
- Antibiotic resistance among bacteria is a growing global challenge. A major reason for this is the limited progress in developing new classes of antibiotics active against Gram-negative bacteria. Here, we investigate the antibacterial activity of a dicationic bisguanidine-arylfuran, originally developed as an antitrypanosomal agent, and new derivatives thereof. The compounds showed good activity (EC50 2-20 µM) against antibiotic-resistant isolates of the Gram-negative members of the ESKAPE group (Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp.) and Escherichia coli with different antibiotic susceptibility patterns, including ESBL isolates. Cytotoxicity was moderate, and several of the new derivatives were less cytotoxic than the lead molecule, offering better selectivity indices (40-80 for several ESKAPE isolates). The molecular mechanism for the antibacterial activity of these molecules is unknown, but sensitivity profiling against human ESKAPE isolates and E. coli collections with known susceptibility patterns against established antibiotics indicates that it is distinct from lactam and quinolone antibiotics.
|
|
| 3. |
- Bourgard, Catarina, 1985, et al.
(författare)
-
Plasmodium vivax Biology: Insights Provided by Genomics, Transcriptomics and Proteomics.
- 2018
-
Ingår i: Frontiers in Cellular and Infection Microbiology. - : Frontiers Media SA. - 2235-2988. ; 8
-
Forskningsöversikt (refereegranskat)abstract
- During the last decade, the vast omics field has revolutionized biological research, especially the genomics, transcriptomics and proteomics branches, as technological tools become available to the field researcher and allow difficult question-driven studies to be addressed. Parasitology has greatly benefited from next generation sequencing (NGS) projects, which have resulted in a broadened comprehension of basic parasite molecular biology, ecology and epidemiology. Malariology is one example where application of this technology has greatly contributed to a better understanding ofPlasmodiumspp. biology and host-parasite interactions. Among the several parasite species that cause human malaria, the neglected Plasmodium vivax presents great research challenges, asin vitroculturing is not yet feasible and functional assays are heavily limited. Therefore, there are gaps in our P. vivax biology knowledge that affect decisions for control policies aiming to eradicate vivax malaria in the near future. In this review, we provide a snapshot of key discoveries already achieved in P. vivax sequencing projects, focusing on developments, hurdles, and limitations currently faced by the research community, as well as perspectives on future vivax malaria research.
|
|
| 4. |
- Chepkirui, Carolyne, et al.
(författare)
-
A new β-hydroxydihydrochalcone from Tephrosia uniflora, and the revision of three β-hydroxydihydrochalcones to flavanones.
- 2022
-
Ingår i: Fitoterapia. - : Elsevier BV. - 1873-6971 .- 0367-326X. ; 158
-
Tidskriftsartikel (refereegranskat)abstract
- The CH2Cl2/MeOH (1:1) extract of the stems of Tephrosia uniflora yielded the new β-hydroxydihydrochalcone (S)-elatadihydrochalcone-2'-methyl ether (1) along with the three known compounds elongatin (2), (S)-elatadihydrochalcone (3), and tephrosin (4). The structures were elucidated by NMR spectroscopic and mass spectrometric data analyses. Elongatin (2) showed moderate antibacterial activity (EC50 of 25.3μM and EC90 of 32.8μM) against the Gram-positive bacterium Bacilus subtilis, and comparable toxicity against the MCF-7 human breast cancer cell line (EC50 of 41.3μM). Based on the comparison of literature and predicted data with that obtained experimentally, we propose the revision of the structure of three β-hydroxydihydrochalcones to flavanones.
|
|
| 5. |
- Chepkirui, Carolyne, et al.
(författare)
-
Benzo[b]naphtho[2,1-d]furans and 2-Phenylnaphthalenes from Streblus usambarensis
- 2023
-
Ingår i: Journal of Natural Products. - : American Chemical Society (ACS). - 0163-3864 .- 1520-6025. ; 86:4, s. 1010-1018
-
Tidskriftsartikel (refereegranskat)abstract
- Three new benzo[b]naphtho[2,1-d]furans, usambarins A–C (1–3), five new 2-phenylnaphthalenes, usambarins D–H (4–8), a new flavan (9), and a new phenyl-1-benzoxepin (10) as well as two known compounds (11 and 12) were isolated from the extract of the stem and roots of Streblus usambarensis (Moraceae). The structures were deduced using NMR spectroscopic and mass spectrometric analyses, and those of compounds 1 and 4 were confirmed by X-ray crystallography. Usambarin D (4) demonstrated moderate antibacterial activity (MIC 9.0 μM) against Bacillus subtilis, while none of the tested compounds were effective against Escherichia coli.
|
|
| 6. |
- Kalenga, Thobias M, et al.
(författare)
-
Antibacterial and cytotoxic biflavonoids from the root bark of Ochna kirkii
- 2021
-
Ingår i: Fitoterapia. - : Elsevier BV. - 1873-6971 .- 0367-326X. ; 151
-
Tidskriftsartikel (refereegranskat)abstract
- The new isoflavonoid kirkinone A (1) and biflavonoid kirkinone B (2) along with six known compounds (3-8) were isolated from the methanolic extract of the root bark of Ochna kirkii. The compounds were identified by NMR spectroscopic and mass spectrometric analyses. Out of the eight isolated natural products, calodenin B (4) and lophirone A (6) showed significant antibacterial activity against the Gram-positive bacterium Bacillus subtilis with MIC values of 2.2 and 28μM, and cytotoxicity against the MCF-7 human breast cancer cell line with EC50 values of 219.3 and 19.2μM, respectively. The methanolic crude extract of the root bark exhibited cytotoxicity at EC50 8.4μg/mL. The isolated secondary metabolites and the crude extract were generally inactive against the Gram-negative Escherichia coli (MIC ≥400μg/mL). Isolation of biflavonoids and related secondary metabolites from O. kirkii demonstrates their chemotaxonomic significance to the genus Ochna and to other members of the family Ochnaceae.
|
|
| 7. |
- Kalenga, Thobias M, et al.
(författare)
-
Biflavanones, Chalconoids, and Flavonoid Analogues from the Stem Bark of Ochna holstii.
- 2021
-
Ingår i: Journal of Natural Products. - : American Chemical Society (ACS). - 1520-6025 .- 0163-3864. ; 84:2, s. 364-372
-
Tidskriftsartikel (refereegranskat)abstract
- Two new biflavanones (1 and 2), three new bichalconoids (3-5), and 11 known flavonoid analogues (6-16) were isolated from the stem bark extract (CH3OH-CH2Cl2, 7:3, v/v) of Ochna holstii. The structures of the isolated metabolites were elucidated by NMR spectroscopic and mass spectrometric analyses. The crude extract and the isolated metabolites were evaluated for antibacterial activity against Bacillus subtilis (Gram-positive) and Escherichia coli (Gram-negative) as well as for cytotoxicity against the MCF-7 human breast cancer cell line. The crude extract and holstiinone A (1) exhibited moderate antibacterial activity against B. subtilis with MIC values of 9.1 μg/mL and 14 μM, respectively. The crude extract and lophirone F (14) showed cytotoxicity against MCF-7 with EC50 values of 11 μg/mL and 24 μM, respectively. The other isolated metabolites showed no significant antibacterial activities (MIC > 250 μM) and cytotoxicities (EC50 ≥ 350 μM).
|
|
| 8. |
- Maeda, Gasper, et al.
(författare)
-
Oxygenated Cyclohexene Derivatives from the Stem and Root Barks of Uvaria pandensis
- 2021
-
Ingår i: Journal of Natural Products. - : American Chemical Society (ACS). - 0163-3864 .- 1520-6025. ; 84:12, s. 3080-3089
-
Tidskriftsartikel (refereegranskat)abstract
- Five new cyclohexene derivatives, dipandensin A and B (1 and 2) and pandensenols A-C (3-5), and 16 known secondary metabolites (6-21) were isolated from the methanol-soluble extracts of the stem and root barks of Uvaria pandensis. The structures were characterized by NMR spectroscopic and mass spectrometric analyses, and that of 6-methoxyzeylenol (6) was further confirmed by single-crystal X-ray crystallography, which also established its absolute configuration. The isolated metabolites were evaluated for antibacterial activity against the Gram-positive bacteria Bacillus subtilis and Staphylococcus epidermidis and the Gram-negative bacteria Enterococcus raffinosus, Escherichia coli, Paraburkholderia caledonica, Pectobacterium carotovorum, and Pseudomonas putida, as well as for cytotoxicity against the MCF-7 human breast cancer cell line. A mixture of uvaretin (20) and isouvaretin (21) exhibited significant antibacterial activity against B. subtilis (EC50 8.7 μM) and S. epidermidis (IC50 7.9 μM). (8'α,9'β-Dihydroxy)-3-farnesylindole (12) showed strong inhibitory activity (EC50 9.8 μM) against B. subtilis, comparable to the clinical reference ampicillin (EC50 17.9 μM). None of the compounds showed relevant cytotoxicity against the MCF-7 human breast cancer cell line.
|
|
| 9. |
- Maeda, Gasper, et al.
(författare)
-
Polyoxygenated cyclohexene derivatives and flavonoids from the leaves of Uvaria pandensis.
- 2022
-
Ingår i: Fitoterapia. - : Elsevier BV. - 1873-6971 .- 0367-326X. ; 158
-
Tidskriftsartikel (refereegranskat)abstract
- Three new oxygenated cyclohexene derivatives, pandensenol D - F (1-3), two new flavanoids, pandensone A and B (4-5), and seven known compounds (6-12) were isolated from the methanol extract of the leaves of Uvaria pandensis Verdc. (Annonaceae). The structures were characterized by NMR spectroscopic and mass spectrometric analyses. The isolated metabolites were evaluated for their antibacterial activity against the Gram-positive bacteria Bacillus subtilis and Staphylococcus epidermidis and the Gram-negative bacteria Enterococcus raffinosus, Escherichia coli, Paraburkholderia caledonica, Pectobacterium carotovorum and Pseudomonas putida, and for cytotoxicity against the MCF-7 human breast cancer cell line. Out of the tested compounds, pandensenol D (1) and (6',7'-dihydro-8'α,9'β-dihydroxy)-3-farnesylindole (12) showed weak whereas (8'α,9'β-dihydroxy)-3-farnesylindole (11) strong activity against B. subtilis. Four of the isolated compounds (1, 4, 11 and 12) showed moderate cytotoxicity against MCF-7 breast cancer cells (EC50>100μM).
|
|
| 10. |
- Tomaz, Kaira C P, et al.
(författare)
-
Identification of potential inhibitors of casein kinase 2 alpha of Plasmodium falciparum with potent in vitro activity.
- 2023
-
Ingår i: Antimicrobial Agents and Chemotherapy. - 0066-4804. ; 67:11
-
Tidskriftsartikel (refereegranskat)abstract
- Drug resistance to commercially available antimalarials is a major obstacle in malaria control and elimination, creating the need to find new antiparasitic compounds with novel mechanisms of action. The success of kinase inhibitors for oncological treatments has paved the way for the exploitation of protein kinases as drug targets in various diseases, including malaria. Casein kinases are ubiquitous serine/threonine kinases involved in a wide range of cellular processes such as mitotic checkpoint signaling, DNA damage response, and circadian rhythm. In Plasmodium, it is suggested that these protein kinases are essential for both asexual and sexual blood-stage parasites, reinforcing their potential as targets for multi-stage antimalarials. To identify new putative PfCK2α inhibitors, we utilized an in silico chemogenomic strategy involving virtual screening with docking simulations and quantitative structure-activity relationship predictions. Our investigation resulted in the discovery of a new quinazoline molecule (542), which exhibited potent activity against asexual blood stages and a high selectivity index (>100). Subsequently, we conducted chemical-genetic interaction analysis on yeasts with mutations in casein kinases. Our chemical-genetic interaction results are consistent with the hypothesis that 542 inhibits yeast Cka1, which has a hinge region with high similarity to PfCK2α. This finding is in agreement with our in silico results suggesting that 542 inhibits PfCK2α via hinge region interaction.
|
|
