In the current work, a new series of novel Δ2‐pyrazoline‐1H‐1,2,3‐triazole derivatives (6a‐6j and 7a‐7f) were synthesized, followed by in situ biological evaluation to assess their antiproliferative and immunomodulatory potential against breast cancer cells (MDA‐MB‐231 and MCF‐7), HUVECs, and PBMCs. The compounds demonstrated antiproliferative effects with IC50 values ranging from 116.92 to 549.73 µM. Seven compounds (6c, 6f, 6h, 6i, 7b, 7e, and 7f) exhibited promising antiproliferative activities with IC50 values below 140 µM in both MDA‐MB‐231 and MCF‐7 cancer cell lines. Compound 7f demonstrated the highest antiproliferative effect with IC50 values of 116.92 µM in MDA‐MB‐231 and 124.72 µM in MCF‐7, with an acceptable cytotoxicity profile in normal HUVEC cells (IC50 = 208.41 µM); thus, immunomodulatory effects of 7f on checkpoint signaling were further evaluated. Compound 7f showed a dose‐dependent increase of TIGIT, PD‐1, and LAG‐3 expression in CD3+ T cells. These changes may enhance responsiveness to checkpoint‐targeted therapies while reflecting complex regulation of T‐cell function. In silico target fishing and molecular docking reflect calpain as a probable target for 7f, while DFT analysis indicates electrophilic and nucleophilic positions within 7f may help its interaction with the target. Molecular dynamics (MD) simulation supports strong binding of 7f with binding energy (−22.259 ± 4.71 kcal mol−1).
S. Khan, H. Moghtaderi, Ebrahimi Amirhossein et al.· Chemistry and Biodiversity· 0 citations
Cotton leaf curl disease (CLCuD), caused by begomoviruses and their betasatellites, is a major threat to cotton production, especially in South Asia, where periodic viral outbreaks continue to affect cotton yields, quality, and livelihoods. The advent of CRISPR/Cas9 gene-editing technology has transformed plant biotechnology, offering efficient, accurate, and programmable methods for combating viral pathogens at the genetic level. Here, the antiviral efficacy of two most widely used CRISPR/Cas9 binary plant expression vectors, pKSE401 and pHSE401, was tested in Nicotiana benthamiana against Cotton leaf curl Kokhran virus (CLCuKoV) and Cotton leaf curl Multan betasatellite (CLCuMuB). The guide-RNAs (gRNAs) were designed to target viral genes that play significant roles in pathogenicity and replication, with pHSE401 encoding a single gRNA and pKSE401 a multiplex of two gRNAs. Agrobacterium-mediated transient transformation and viral inoculation experiments revealed that both CRISPR/Cas9 vectors effectively delayed symptom onset and reduced virus titers relative to infected controls. Remarkably, the multiplex pKSE401 system was more effective at suppressing viral infection, achieving about a 90% reduction in viral accumulation compared with a 75% reduction by the single gRNA pHSE401 construct. pKSE401-treated plants showed delayed symptom development, reduced severity, and partial recovery, demonstrating the improved efficiency of multiplex genome editing. The results demonstrate the cutting-edge potential of CRISPR/Cas9 multiplex approaches as next-generation methods for designing sustainable resistance to multifaceted plant virus diseases. This research not only contributes to our understanding of CRISPR-based antiviral response mechanisms but also provides a promising avenue for designing broad-spectrum, sustainable resistance against viral epidemics in cotton and other commercially valuable crops.
Farwa Yaqub, Sidra Ashraf, Ahmed Al‐Harrasi et al.· Plant Protection· 0 citations