Structure-based drug discovery of novel fused-pyrazolone carboxamide derivatives as potent and selective AXL inhibitors

As a novel and promising antitumor target, AXL plays an important role in tumor growth, metastasis, immunosuppression and drug resistance of various malignancies, which has attracted extensive research interest in recent years. In this study, by employing the structure-based drug design and bioisosterism strategies, we designed and synthesized in total 54 novel AXL inhibitors featuring a fused-pyrazolone carboxamide scaffold, of which up to 20 compounds exhibited excellent AXL kinase and BaF3/TEL-AXL cell viability inhibitions. Notably, compound 59 showed a desirable AXL kinase inhibitory activity (IC50: 3.5 nmol/L) as well as good kinase selectivity, and it effectively blocked the cellular AXL signaling. In turn, compound 59 could potently inhibit BaF3/TEL-AXL cell viability (IC50: 1.5 nmol/L) and significantly suppress GAS6/AXL-mediated cancer cell invasion, migration and wound healing at the nanomolar level. More importantly, compound 59 oral administration showed good pharmacokinetic profile and in vivo antitumor efficiency, in which we observed significant AXL phosphorylation suppression, and its antitumor efficacy at 20 mg/kg (qd) was comparable to that of BGB324 at 50 mg/kg (bid), the most advanced AXL inhibitor. Taken together, this work provided a valuable lead compound as a potential AXL inhibitor for the further antitumor drug development.

Effect of acute hypervolemic hemodilution with 6% hydroxyethyl starch 130/0.4 on pharmacodynamics of propofol during successful laryngeal mask airway implantation / 中华麻醉学杂志

Objective:To investigate the effect of acute hypervolemic hemodilution (AHH) with 6% hydroxyethyl starch 130/0.4 on pharmacodynamics of propofol during successful laryngeal mask airway (LMA) implantation.Methods:American Society of Anesthesiology physical status Ⅰ or Ⅱ patients, aged 30-60 yr, with body mass index of 18.5-25.0 kg/m 2, undergoing elective extensive total hysterectomy under general anesthesia, were divided into 2 groups: AHH group (group A) and control group (group C). In group A, 6% hydroxyethyl starch 130/0.4 was infused at a rate of 20 ml/min for AHH, and the target hematocrit was 30%.In group C, lactated Ringer′s solution was infused according to the " 4-2-1" rule to supplement physiological requirements, and anesthesia induction was performed after 10 min of stabilization.Sufentanil was administered by target-controlled infusion using Bovil pharmacokinetic model with effect-site concentration (Ce) of 0.25 ng/ml, 3 min later propofol was given by target-controlled infusion using Schnider model.The Ce of propofol in the first patient was set at 5.0 μg/ml.Each time the concentration of propofol was increased/decreased by 0.5 μg/ml according to the sequential method.LMA was inserted following 1 min equilibration between plasma concentration and Ce of propofol.The trial was terminated when 8 consecutive inflection points of failed/successful LMA insertion occurred.The EC 5, EC 50, EC 95 and 95% confidence interval (95% CI) of propofol were calculated by probit regression analysis. Results:In group A, the EC 5 (95% CI), EC 50 (95% CI) and EC 95 (95% CI) of propofol when LMA was successfully placed were 4.237 (3.090-4.514) μg/ml, 4.802 (4.500-5.078) μg/ml and 5.443 (5.125-7.304) μg/ml, respectively.In group C, the EC 5 (95% CI), EC 50 (95% CI) and EC 95 (95% CI) of propofol when LMA was successfully placed were 2.408 (1.190-2.756) μg/ml, 3.120 (2.690-3.472) μg/ml and 4.042 (3.582-7.431) μg/ml, respectively.There was significant difference in EC 5, EC 50 and EC 95 between the two groups ( P<0.01). Conclusion:AHH with 6% hydroxyethyl starch 130/0.4 can decrease the efficacy of propofol when LMA is successfully implanted.

The effect of preoperative anxiety on propofol EC50 for no-movement during painless gastroscopy / 中国医师杂志

Objective To investigate the effect of preoperative anxiety on propofol EC50 for nomovement during Painless gastroscopy.Methods Thirty-one patients (without anxiety) and twenty-seven patients (with anxiety) undergoing gastroscopy under general anesthesia were enrolled on the study.Anesthesia was conducted with a target-controlled infusion (TCI) of propofol.The initial target effect-site propo fol concentration (Ceprop) was 5.0 μg/ml and was adjusted stepwise by 0.5 μg/ml by an up-down sequential method to reach no-movement.Results Propofol EC50 to induce no-movement was higher in patients with anxiety than those without anxiety (5.32 μg/ml,95％ CI:4.75-5.88 μg/ml vs 4.75 μg/ml,95％ CI:4.48-5.01 μg/ml,P ＜ 0.05).Conclusions During painless gastroscopy,patients with anxiety had a higher propofol EC50 for no-movement compared with those without anxiety when intravenous injected of fentanyl 0.1 μg/kg.

Effect of Transcranial Direct Current Stimulation on Regional Homogeneity After Sleep Deprivation by fMRI / 中国医学影像学杂志

Purpose To observe the impact of sleep deprivation (SD) for 24 hours on human brain regional homogeneity (ReHo) by using functional magnetic resonance imaging (fMRI),and briefly discuss the intervening effect of transcranial direct current stimulation (tDCS) on ReHo after SD.Material and Methods Sixteen healthy individuals were enrolled,and self-controlled study were adopted.Resting-state fMRI scans were performed in wakefulness,after SD,and after tDCS (true or placebo stimuli).The ReHo analysis approach was employed to calculate the ReHo values of whole brain in different states.The differences between before and after SD,and between true and false tDCS after SD,were analyzed.Results Compared with wakefulness,SD mainly enhanced ReHo in brain regions of left precentral gyrus,right precentral gyrus,bilateral temporal lobe and thalamus,but decreased ReHo in brain regions of right precuneus,left superior occipital gyrus,limbic lobe and bilateral angular gyrus,superior frontal gyrus,middle frontal gyrus.Compared with placebo stimuli,the true tDCS enhanced ReHo in brain regions of bilateral precuneus,angular gyrus and right forehead,superior middle gyrus,but decreased ReHo in brain regions of the medial side of left frontal lobe,right precentral gyrus,parahippocampal gyrus,substantia nigra and bilateral temporal lobe,pons and so on.Conclusion SD can cause ReHo change in a large number of brain regions.The tDCS on the prefrontal cortex improves the brain regions with ReHo reduction caused by SD.The improved brain regions mainly include bilateral precuneus/posterior cingulate cortex.Therefore,tDCS on the prefrontal cortex can improve brain dysfunction caused by SD.

Effect of sleep deprivation on learning and memory network in healthy adults by fMRI / 安徽医科大学学报

Objective To observe the neural mechanism of learning and memory network disturbance induced by 36 hours sleep deprivation(SD).Methods 18 young male healthy volunteers were enrolled in this study.Resting state functional magnetic imaging(rs-fMRI) data was collected by blood oxygen level dependence functional magnetic imaging(BOLD-fMRI) during rest wakefulness(RW) and 36 hours SD.Bilateral hippocampus were selected as seed points by REST software automated anatomical labeling template.And then compared the differences of functional connectivity(FC) between bilateral hippocampus and whole brain regions by using paired-t test within statistical parametric mapping 8 (SPM8) software.Finally,correlation analysis had been done between significant FC of brain regions and WMS MQ values (SD-RW).Results Compared with RW,the bilateral hippocampus showed positive connected with the right dorsal posterior cingulate cortex,and showed negative functional connectivity with bilateral postcentral gyrus.The WMS MQ was significantly decreased after 36 hours SD.The connection between bilateral hippocampus and the right posterior cingulate cortex showed negative correlation with the deductionofWMS MQ (left:r=-0.463,P=0.015;right:r=-0.546,P=0.003).Conclusion SD has a significant effect on bilateral hippocampal FC.The FC between bilateral hippocampus and right dorsal posterior cingulate cortexis significant increased,while between bilateral postcentral gyrus is decreased after 36 hours SD.The former may be an important neurological mechanism in learning and memory ability regulation after 36 hours sleep deprivation.

Effects of light acute hypervolemic hemodilution on early postoperative cognitive function in elderly patients / 实用医学杂志

Objective To investigate the effects of light acute hypervolemic hemodilution on early postop-erative cognitive function in elderly patients. Methods A total of 60 patients treated by radical surgery were equally randomized to an AHH and a non-AHH control group.Using the Mini-Mental State Examination scale (MMSE),we evaluated the cognitive function of the patients.We made comparisons between the two groups in the plasma S100β levels at T0(before anesthesia induction),T1(immediately after hemodilution),T2(immediately after operating),T(36 hour after operating)and T(424 hour after operating).Results The S-100β lever at T2、T3 and T4were markedly higher than T0in both groups(P<0.05);At T3and T4,the S-100β lever was higher in non-AHH control group than AHH group(P<0.05).There was no significant difference in the incidence of postopera-tive cognitive dysfunction between the two groups(P>0.05)There was no significant difference in the incidence of postoperative cognitive dysfunction between the two groups(P > 0.05). Conclusions AHH can significantly reduce plasma S100β in elderly patients,but there is no effect on early postoperative cognitive function.

The influence of preoperative anxiety on popofol EC50 for no-movement during gastroscopy / 实用医学杂志

Objective To investigate whether the median(50%)effective effect-concentration(EC50)of propofol inducing loss of consciousness (LOC) varies. Methods 56 patients undergoing gastroscopy under general anaesthesia were enrolled on the study. Anaesthesia was conducted with a target-controlled infusion(TCI) of propofol. The initial target effect-site propofol concentration (Ceprop) was 5.00 μg/mL and was adjusted stepwise by 0.50μg/mL by an up-down sequential method to reach no-movement. Results Propofol EC50 to induce no-movement was higher in patients with anxiety than those without anxiety(6.46μg/mL vs. 5.75μg/mL,P<0.05). Conclusions During general anaesthesia ,patients with anxiety had a higher propofol EC50 for no-movement compared with those without anxiety. Differences in preoperative anxiety levels may reduce anaesthetic effects.

The influence of preoperative anxiety on popofol EC50 for no-movement during gastroscopy / 实用医学杂志

Objective To investigate whether the median(50%)effective effect-concentration(EC50)of propofol inducing loss of consciousness (LOC) varies. Methods 56 patients undergoing gastroscopy under general anaesthesia were enrolled on the study. Anaesthesia was conducted with a target-controlled infusion(TCI) of propofol. The initial target effect-site propofol concentration (Ceprop) was 5.00 μg/mL and was adjusted stepwise by 0.50μg/mL by an up-down sequential method to reach no-movement. Results Propofol EC50 to induce no-movement was higher in patients with anxiety than those without anxiety(6.46μg/mL vs. 5.75μg/mL,P<0.05). Conclusions During general anaesthesia ,patients with anxiety had a higher propofol EC50 for no-movement compared with those without anxiety. Differences in preoperative anxiety levels may reduce anaesthetic effects.

Long non-coding RNA H19 promoted lung cancer cells migration and invasion through miR-107 / 中国免疫学杂志

Objective:To investigate the effect and the related mechanism of H19 on the invasion and migration ability of pancreatic cancer cells.Methods: The expression of H19 in tissues of pancreatic carcinoma and non-carcinoma adjacent tissues of pancreatic carcinoma were detected.qPCR was used to detect the expression of H19 in different pancreatic cancer cells.The migration and invasion ability of pancreatic cancer cells was detected after silencing H19 using wound healing assays and Transwell matrigel invasion assays.Dual-Luciferase Reporter Assay System was used to detect miR-107regulating H19.The expression of miR-107 in tissues of pancreatic carcinoma and non-carcinoma adjacent tissues of pancreatic carcinoma were detected.The regulation of miR-107 on the migration and invasion ability of pancreatic cancer cells were detected after silencing H19 using wound healing assays and Transwell matrigel invasion assays.Subcutaneous tumor was used to detect the size and volume of the tumor after inject the tumor cells.Immunohistochemistry was used to detect the expression of Ki67 and PCNA.Results: Compared with non-carcinoma adjacent tissues of pancreatic carcinoma,the expression of H19 of tissues of pancreatic carcinoma was significantly increased.The expression of H19 in pancreatic cancer cell A549 was highest.Silencing H19 could inhibit the migration of human pancreatic cancer A549 cells.miR-107 was the direct target of H19.Compared with non-carcinoma adjacent tissues of pancreatic carcinoma,the expression of H19 of tissues of pancreatic carcinoma was significantly decreased.After silencing H19,inhibiting the expression of miR107 could inhibit the migration of human pancreatic cancer A549 cells.Compared with the group of H19-siRNA,H19-siRNA+miR-107-inhibitor group mice tumor volume and weight were significantly bigger.Immunohistochemistry showed that compared with H19-siRNA group,the expression Ki67 and PCNA expression in H19-siRNA+miR-107-inhibitor group increased.Conclusion: H19 plays the role of tumor promotor factor in pancreatic cancer.H19 can affect the invasion and migration ability of pancreatic carcinoma cells by regulating miR-107.

Construction and sequence analysis of a drought-induced full-length cDNA library from Ammopiptanthus mongolicus / 生物工程学报

Ammopiptanthus mongolicus shows very strong resistance to severe environments. To isolate drought-resistant genes and elucidate drought-resistant molecular mechanisms of the plant, we constructed a drought-induced full-length cDNA library using SMART (Switching mechanism at 5'-end of RNA transcript) technique. The phage titer of the unamplified library was 1.6 x 10(7) PFU/mL; the recombination percentage was 97.7%; and the sizes of most cloned cDNA fragments were around 1 kb. Three thousand positive clones were randomly selected and sequenced from their 5' ends, and a total of 1 450 Unigenes were identified. By Blast searches against the Nt, Nr and Swissprot databases, we found that 919 Unigenes (amount to 63.4%) showed significant similarity to the annotated genes, and the remaining 531 Unigenes (amount to 36.6%) represented novel genes without any annotation. Among the functional categories of the GO (Gene Ontology) classification, the terms related to physiological process, cellular process, binding, catalytic activity and cellular components were dominant. The next abundant terms were for organelle, protein complex, transporter activity and structural molecule activity. In addition, there were a significant proportion of the terms involved in stimulus response, gene expression regulation, regulation of physiological and biochemical processes and signal transduction. Many of the annotated Unigenes were found to be related to plant resistance to abiotic stresses, and expression analyses of 6 out of these genes by semi-quantitive RT-PCR confirmed their involvements in the response of A. mongolicus to drought stress. These results laid a foundation for the expression profile analysis and the cloning and characterization of drought-resistant genes from the plant in the future.