Combinatorial Chemistry & High Throughput Screening - Volume 22, Issue 1, 2019

Aim and Objective: Avicularin has been found to inhibit the proliferation of HepG-2 cells in vitro in the screening of our laboratory. We intended to explain the molecular mechanism of this effect. Therefore, the combined methods of reverse molecular docking and network pharmacology were used in order to illuminate the molecular mechanisms for Avicularin against cancer. Materials and Methods: Potential targets associated with anti-tumor effects of Avicularin were screened by reverse molecular docking, then a protein database was established through constructing the drugprotein network from literature mining data, and the protein-protein network was built through an in-depth exploration of the relationships between the proteins, and then the network topology analysis was performed. Additionally, gene function and signaling pathways were analyzed by Go bio-enrichment and KEGG Pathway. Results: The result showed that Avicularin was closely related to 16 targets associated with cancer, and it may significantly influence the pro-survival signals in MAPK signaling pathway that can activate and regulate a series of cellular activities and participate in the regulation of cell proliferation, differentiation, transformation and apoptosis. Conclusion: The network pharmacology strategy used herein provided a powerful means for the mechanisms of action for bioactive ingredients.

Objective: Gallstone formation is a pathological process of mineralization in the human body. Determination of the morphology and ultrastructure of gallstones holds the key to understanding the pathophysiology of gallbladder disease. Synchrotron radiation phase-contrast Xray microtomography is a novel technology, which is designed for comprehensive analysis of gallstone ultrastructure. Materials and Methods: Nine human gallstones were obtained from the Department of Pathology, Qingpu branch of Zhongshan Hospital Affiliated to Fudan University (China), and scanned by synchrotron radiation μCT (SR μCT). The imaging data generated by SR μCT scan were analyzed. Results: The three-dimensional ultrastructure of human gallstones corresponding to their cholesterol and bile pigment composition was determined. Conclusions: The ultrastructure of gallstones exhibits considerable diversity and complexity. The synchrotron radiation phase-contrast X-ray microtomography is a valuable tool for in-depth study of human gallstones.

Aim and Objective: In this study, biological synthesis of cobalt nanoparticles was developed in the presence of ginger extract as the reducing and capping agent through the simple and convenient co-precipitation method. Materials and Methods: The as-synthesized cobalt nanoparticles were characterized by X-ray diffraction (XRD), scanning Electron Microscopy (SEM), spectra energy dispersive analysis of Xray (EDS), Fourier transform infrared (FT-IR), and vibrating sample magnetometer (VSM) techniques. According to the vibrating sample magnetometer, cobalt nanoparticles show paramagnetic behaviour at room temperature. Furthermore, the effect of ginger extract concentration on the UV-Vis absorbance of Co nanoparticles was investigated. Based on the UVVis absorbance spectra, increasing ginger extract concentration causes particle size to decrease. In addition, the catalytic performance of the synthesized cobalt nanoparticles was investigated in the preparation of pyrano[2,3-c]pyrazoles via one-pot four-component reactions of aryl aldehydes, hydrazine hydrate, malononitrile and diethyl acetylenedicarboxylate. Result and Conclusion: The prepared pyrano[2,3-c]pyrazole derivatives were obtained in high yields within short reaction times and the nanocatalyst was easily separated using an external magnet and reused for several times with no significant loss of its activity.

Aim: To analyse the clinical outcomes of patients with lung cancer treated with first and multiple-line chemotherapy and tyrosine kinase inhibitor (TKI). Patients & Methods: The present study included a total of 89 patients of whom lung cancer was histologically confirmed between July 2016 and September 2017. Patients’ demographics, chemotherapy/TKI treatment details and clinical outcomes were retrieved. The progression-free survivals (PFS) after first-line and multiple-line treatments were evaluated using Kaplan-Meier analysis with log-rank test. Risk factors for progressive disease (PD) were identified using Cox multivariate regression model. Results: A total of 50 males and 39 females were enrolled. About 90% of the tumors were histologically classified as adenocarcinoma, and 77/89 cases (86.5%) were at TNM stage IV. The median PFS for the first-line treatment was 6.17 months. After first-line treatment, more favourable PFS was observed in patients who had prior surgery of lung cancer (P = 0.002). Multivariate analysis showed that patients who had received no prior surgical treatment for lung cancer were at higher risk of PD (OR, 4.311; 95% CI, 1.836 to 10.120; P = 0.0008). Besides, in patients with driver mutations, those who received no TKI treatment were under higher risk of PD compared to those who had been treated with TKI (OR, 4.876; 95% CI, 1.877 to 12.666; P = 0.0011). The median PFS for the multiple-line treatment was 24.67 months. After multiple-line treatments, favourable PFS was associated with tumor histological types of adenocarcinoma (P = 0.041), genetic lesions at exon 19 of EGFR (P = 0.001) and fourth-line treatment (P = 0.001). Risk factors for PD after multiple-line treatments were no prior surgery for lung cancer (OR, 3.335; 95% CI, 1.158 to 9.605; P = 0.0256), no TKI use in multiple-line treatment (OR, 10.095; 95% CI, 2.405 to 42.378; P = 0.0016), and being treated by first-line treatment alone (OR, 30.421; 95% CI, 4.813 to 192.269; P = 0.0003). Conclusion: The present study demonstrated the benefits of TKI in patients with advanced lung cancer, providing insights into the refinement of the management strategy.

Aim and Objective: Small molecule targeted drugs can effectively reduce the toxicity and side effects of drugs, and improve the efficacy of drugs by their specific antitumor activity. Hence, the development of small molecular targeted drugs for cancer has important significance. This study was undertaken to design and synthesize novel phenazine-chromene hybrid molecules in order to optimize the structure and improve the efficacy of this kind of hybrids. Materials and Methods: O-diaminobenzene was used as starting material to synthesize twentyfour heterocyclic compounds designed as hybrid molecules of phenazine and 4H-chromene pharmacophores by facile methods. The structures of the compound were confirmed by 1H NMR, 13C NMR and HRMS. Furthermore, the synthesized compounds were evaluated for in vitro activity against four human cancer cell lines and two non-cancer cell lines by MTT test. Results: Some compounds showed strong cytotoxic activities against HepG2 and A549 cancer lines (IC50 = 5-10 μM). Comparing 2i with 2l, the introduction of hydrophilic groups on the phenazine core could not improve the antiproliferative activity significantly. Except 2d and 3c, compounds owning chlorine substituent on the 4H-chromene pharmacophore seemingly contribute to enhance the compounds’ antiproliferative activity. Specially, compound 3c showed highest cytotoxicity against A549 cells with IC50 values of 3.3±0.4 μM. Furthermore, all compounds showed low or no cytotoxicity against HUVEC and L02 non-cancer cells in vitro. Conclusion: Compound 3c may be used as potential lead molecule against A549 cancer cells.

Background: Retinopathy of Prematurity (ROP) is a pathophysiologic condition of the retina due to abnormal proliferation of retinal vessels. Objective: The study aimed too ascertain the importance of vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), interleukin-33 (IL-33) and endocan in the diagnosis and follow-up of ROP. Methods: This prospective cohort study was conducted in the neonatal intensive care unit (NICU) of Health Science University, Zekai Tahir Burak Maternity Teaching Hospital, Ankara, Turkey, between February 2017 and August 2018. Preterm infants (gestational age (GA) of ≤32 weeks and birth weight of ≤1500 gr), diagnosed ROP were included in the study. VEGF, IGF-1, IL-33 and endocan levels were evaluated in the cord blood and in the serum before and after treatment of infants in the ROP and control groups. Results: A final number of 146 infants were included in the study. During the study period, 73 infants were identified as the ROP group, and 73 infants were allocated as the control group. In the ROP group, the cord blood VEGF value was higher than the control group (p <0.05). However, IGF-1 levels in the cord blood were lower in the ROP group than control (P<0.05). IL-33 and endocan values in the cord blood were similar in both control and ROP groups (p>0.05). Although serum levels of IL-33, VEGF and endocan were higher before laser treatment, these biomarkers decreased significantly after laser treatment (p <0.05). Conclusion: We determined that serum IL-33 and endocan levels might be suggested as sensitive novel markers for the prediction of severe ROP.

Aim and Objective: Human full-length cyclic nucleotide phosphodiesterase isozyme 4B2 (hPDE4B2) as the target for screening and characterizing inhibitors suffers from low activity yield and the coexistence of two conformational states bearing different affinities for (R)-rolipram. Hence, the 152~528 truncate of hPDE4B2 existing only in the low-affinity conformation state for (R)-rolipram was compared against the full-length hPDE4B2 to characterize inhibitors. Materials and Methods: With 6His-SUMO tag at the N-terminus, both the full-length hPDE 4B2 (SF-hPDE4B2) and the 152~528 truncate (ST-hPDE4B2) were expressed in Escherichia coli cells, purified through Ni-NTA column and compared for the characterization of inhibitors. The inhibition constants (Ki) of some synthesized rolipram analogues against both targets were determined with 96-well microplate through the coupled action of monophosphatase on AMP and spectrophotometric assay of phosphate with malachite green. Results: After affinity purification with Ni2+-NTA column, ST-hPDE4B2 showed about 30-fold higher specific activity and 100-fold higher activity yield than SF-hPDE4B2; Ki of (R)-rolipram on ST-hPDE4B2 was consistent with that on the low-affinity state of the untagged full-length hPDE4B2 expressed in insect cells. Of some representative rolipram analogues as inhibitors, a dual-logarithm model quantitatively described their monotonic association, and Ki from 0.010 mM to 8.5 mM against SF-hPDE4B2 was predicted from Ki against ST-hPDE4B2, supporting the discovery of consistent hits by the use of both targets with a pair of properly-set cutoffs. Conclusion: ST-hPDE4B2 with much higher activity yield may be a favorable alternative target to characterize/screen rolipram analogues as hPDE4B inhibitors in high-throughput mode.

Background: HIV infections are a worldwide health problem. HIV infection reduces CD4+ cell counts. Oxidative stress might play an important role in the stimulation of virus replication and immunodeficiency. Saliva might be the first line of defense against oxidative stress. Objective: The aim of this study was to evaluate the oxidative stress marker and antioxidant levels of saliva in HIV-infected patients by measuring total antioxidant capacity and malondialdehyde level. Methods: A total of 49 HIV-positive patients and 49 healthy HIV-negative individuals were randomly selected. All the patients were clinically examined. Five mL of unstimulated whole saliva was collected and evaluated by spectrophotometric assay. Data were analyzed with STATA 11. Results: Mean ages of the case and control groups were 28 and 33 years, respectively. Salivary malondialdehyde levels were significantly higher in the HIV-positive group (3.68±2.26) compared to the healthy control group (2.79±1.91). Levels of salivary total antioxidant capacity were significantly lower in the HIV-positive group (0.20± 0.09) compared to the control group (0.27±0.10). Conclusion: The antioxidant defense system in HIV-positive individuals was low and oxidative stress was high in this population. Saliva might be used as a diagnostic tool for antioxidant changes in HIV-positive patients in the future. There were changes in salivary antioxidant defense system and oxidative stress in HIV-positive individuals. Antioxidant supplements might help local salivary and general health statuses.

Background: Epilepsy is a serious clinical condition characterized by recurrent seizures. Oxidative stress plays an important role in the etio-pathogenesis of epilepsy. Measurements of serum thiol and disulfide levels were used to evaluate the antioxidant status of the body. Objective: The aim of this study was to determine serum levels of thiol and disulfide in epileptic pediatric patients. Methods: Ninety patients, 54 epilepsy and 36 controls were included in the study. Serum levels of native thiol total thiol and disulfide were measured and disulfide/native, disulfide / total thiol and native thiol/ total thiol ratios were calculated. Hence, the ratios of disulfide/ native thiol, disulfide / total thiol and native thiol/ total thiol were calculated. Results: Serum levels of native thiol, total thiol and disulfide were significantly lower in the epilepsy group than the control group. The ratio of disulfide/native thiol and disulfide / total thiol were significantly higher in the study group than the control group. As well as, the native thiol / total thiol ratio was lower in the epilepsy group than the control group. Native thiol, total thiol and disulfide were significantly lower in the epilepsy group who were taking anti-epileptic drugs than those who were not taking anti-epileptic drugs. Conclusion: We demonstrated a meaningful relationship between oxidative stress markers and epilepsy in pediatric patients.

Background: Tissue fibrosis increases in the structure of the atrial tissue of atrial fibrillation patients. Prolidase enzyme regulates collagen synthesis. There may be an association between electrocardiography (ECG) findings and prolidase activity. Objective: This study investigated the association between atrial conduction time and prolidase activity, a collagen synthesis enzyme, and P-wave dispersion (PWD) in patients with Paroxysmal Atrial Fibrillation (PAF). Methods: Exclusion criteria included the age of <18 years, heart failure, diabetes, hypertension, hyperlipidemia, malignancy, cerebrovascular disease, chronic respiratory distress, osteoporosis, rheumatoid arthritis, renal disease, cirrhosis, and other types of arrhythmia. Patients diagnosed with PAF within 48 hours were considered to have a definite diagnosis. PWD was calculated using a 12-lead ECG, and inter- and intraatrial electromechanical delay (EMD) was assessed using tissue Doppler imaging and conventional echocardiography. Serum prolidase levels were measured in both groups. Results: A total of 43 patients with PAF (20 female, 23 male; mean age, 46.8 ± 5.7 years) and 42 healthy volunteers (21 female, 21 male; mean age, 43.9 ± 5.1 years) were included in the study. Inter- and intraatrial EMD, PWD, minimum P-wave (Pmin), and maximum P-wave (Pmax) measurements were significantly higher (39.7 ± 2.7, 35.7 ± 2.3, p < 0.001; 13.2 ± 2.6, 8.5 ± 1.9, p < 0.001; 47.1 ± 11, 24.1 ± 7.1, p < 0.001; 69.8 ± 8.8, 66.7 ± 10.2, p < 0.130; 114.8 ± 13, 93.6 ± 8.6, p < 0.001, respectively) and serum prolidase levels were significantly lower in patients with PAF compared to healthy controls (3.96 ± 1.2, 8.5 ± 3.56, p < 0.001). In patients with PAF, correlation analysis showed a negative correlation between prolidase levels and intra- and interatrial EMD, PWD, and Pmax (r = -0.41, p < 0.05; r = -0.54, p < 0.05; r = -0.62, p < 0.05; r = -0.49, p < 0.05, respectively). Interatrial EMD showed a significant positive correlation with intraatrial EMD, Pmax, and PWD in patients with PAF (r = 0.90, p < 0.05; r = 0.574, p < 0.05; r = 0.43, p < 0.05, respectively). Additionally, the level of high-sensitivity C-reactive protein (hs-CRP) was significantly higher in patients with PAF (6.6 ± 8, 1.8 ± 1.6, p < 0.001). Conclusion: The decreased plasma prolidase activity in patients with PAF may explain the irregularity of the collagen metabolism of different extracellular components and may indicate the onset of atrial remodeling. Changes in PWD, interatrial EMD, and serum prolidase level may predict PAF before diagnosis.