Synthesis and cellular characterization of novel isoxazolo- and thiazolohydrazinylidene-chroman-2,4-diones on cancer and non-cancer cell growth and death.

Coumarins are extensively studied anticoagulants that exert additional effects such as anticancerogenic and even anti-inflammatory. In order to find new drugs with anticancer activities, we report here the synthesis and the structural analysis of new coumarin derivatives which combine the coumarin core and five member heterocycles in hydrazinylidene-chroman-2,4-diones. The derivatives were prepared by derivatization of the appropriate heterocyclic amines which were used as electrophiles to attack the coumarin ring. The structures were characterized by spectroscopic techniques including IR, NMR, 2D-NMR and MS. These derivatives were further characterized especially in terms of a potential cytotoxic and apoptogenic effect in several cancer cell lines including the breast and prostate cancer cell lines MCF-7, MDA-MB-231, PC-3, LNCaP, and the monocytic leukemia cell line U937. Cell viability was determined after 48 h and 72 h of treatment with the novel compounds by MTT assay and the 50% inhibitory concentrations (EC50 values) were determined. Out of the 8 novel compounds screened for reduced cell viability, 4c, 4d and 4e were found to be the most promising and effective ones having EC50 values that were several fold reduced when compared to the reference substance 4-hydroxycoumarin. However, the effects were cancer cell line dependent. The breast cancer MDA-MB-231 cells, the prostate cancer LNCaP cells, and U937 cells were most sensitive, MCF-7 cells were less sensitive, and PC-3 cells were more resistant. Reduced cell viability was accompanied by increased apoptosis as shown by PARP-1 cleavage and reduced activity of the survival protein kinase Akt. In summary, this study has identified three novel coumarin derivatives that in comparison to 4-hydroxycoumarin have a higher efficiency to reduce cancer cell viability and trigger apoptosis and therefore may represent interesting novel drug candidates.

Male Infertility and Sperm DNA Fragmentation.

BACKGROUND: One of the main factors affecting male infertility is DNA fragmentation in sperm. Male infertility is a heterogeneous group of disorders, known causes account for only 30-50%, and unknown cause (idiopathic) constitute the rest. Infertility involves nearly 15% of couples in the reproductive age, and only the male problem involves about 40% of the problems. AIM: We have studied our DNA damage to sperm cells of a group of infertile males (113 patients) with abnormal sperm parameters (oligoasthenospermia and oligospermia) and a group of male patients (80 patients) with normal semen parameters (normospermia) to document whether the Sperm Chromatin Dispersion (SCD) analysis could increase the information obtained from the sperm routine analysis to explain the causes of infertility. MATERIALS: A group of 193 patients were analysed, 113 patients in the working group and 80 patients in the control group were screened. The ejaculate samples were taken by the patient to whom the reason for the analysis was explained. All patients were from the Republic of Kosovo. Samples are collected from 2014/2018. Sperm Chromatin Dispersion (SCD) analyses in the ejaculate were analysed by the Biolab Zafi laboratory in Peja. RESULTS: Clinical data were compared between the two groups by one-way ANOVA, mean ± SD, student's t-test. A p-value of less than P < 0.05% was considered statistically significant. Outcomes: In our study, we have gained significant (P < 0.05) results in the workgroup and the control group across all hormonal parameters, sperm parameters, and fragmented DNA in the sperm. CONCLUSION: Based on our obtained results we can conclude that DNA fragmentation in spermatozoa is useful in the selection of unsuitable DNA sperm for use in ART methods. We conclude that our DNA fragmentation analysis results are encouraging and can be used for diagnostic purposes in determining male infertility.