A Facile Ugi/Ullmann Cascade Reaction to Access Fused Indazolo-Quinoxaline Derivatives with Potent Anticancer Activity.

A facile methodology for the construction of a complex heterocycle indazolo-fused quinoxalinone has been developed via an Ugi four-component reaction (U-4CR) followed by an intramolecular Ullmann reaction. The expeditious process features an operationally simple approach, time efficiency, and a broad substrate scope. Biological activity was evaluated and demonstrated that compound 6e inhibits human colon cancer cell HCT116 proliferation with an IC50 of 2.1 µM, suggesting potential applications for developing a drug lead in medicinal chemistry.

Pyrrolyldihydropyrazino[1,2-a]indoletrione Analogue Microtubule Inhibitor Induces Cell-Cycle Arrest and Apoptosis in Colorectal Cancer Cells.

In this study, 2-benzyl-10a-(1H-pyrrol-2-yl)-2,3-dihydropyrazino[1,2-a]indole-1,4,10(10aH)-trione (DHPITO), a previously identified inhibitor against hepatocellular carcinoma cells, is shown to exert its cytotoxic effects by suppressing the proliferation and growth of CRC cells. An investigation of its molecular mechanism confirmed that the cytotoxic activity of DHPITO is mediated through the targeting of microtubules with the promotion of subsequent microtubule polymerisation. With its microtubule-stabilising ability, DHPITO also consistently arrested the cell cycle of the CRC cells at the G2/M phase by promoting the phosphorylation of histone 3 and the accumulation of EB1 at the cell equator, reduced the levels of CRC cell migration and invasion, and induced cellular apoptosis. Furthermore, the compound could suppress both tumour size and tumour weight in a CRC xenograft model without any obvious side effects. Taken together, the findings of the present study reveal the antiproliferative and antitumour mechanisms through which DHPITO exerts its activity, indicating its potential as a putative chemotherapeutic agent and lead compound with a novel structure.

DMAPT­D6 induces death­receptor­mediated apoptosis to inhibit glioblastoma cell oncogenesis via induction of DNA damage through accumulation of intracellular ROS.

Glioblastoma (GBM) is an aggressive malignancy with a high rate of tumor recurrence after treatment with conventional therapies. Parthenolide (PTL), a sesquiterpene lactone extracted from the herb Tanacetum parthenium or feverfew, possesses anticancer properties against a wide variety of solid tumors. In the present study, a series of PTL derivatives were synthesized and screened. An inhibitor, dimethylaminoparthenolide (DMAPT)­D6, a derivative of the PTL prodrug DMAPT in which the hydrogen of the dimethylamino group is substituted for the isotope deuterium, induced significant cytotoxicity in GBM cells in vitro and induced cell cycle arrest at the S­phase in a dose­dependent manner. Furthermore, mechanistic investigation indicated that through increasing the levels of intracellular accumulation of reactive oxygen species (ROS), DMAPT­D6 triggered DNA damage and finally death receptor­mediated extrinsic apoptosis in GBM cells, suggesting that DNA damage induced by DMAPT­D6 initiated caspase­dependent apoptosis to remove damaged GBM cells. Taken together, these data suggested that ROS accumulation following treatment with DMAPT­D6 results in DNA damage, and thus, death­receptor­mediated apoptosis, highlighting the potential of DMAPT­D6 as a novel therapeutic agent for the treatment of GBM.

A Novel Imidazopyridine Derivative Exhibits Anticancer Activity in Breast Cancer by Inhibiting Wnt/ß­catenin Signaling.

BACKGROUND: Breast cancer exhibits poor prognosis and high relapse rates following chemotherapy therapeutics. Thus, this study aims to develop effective novel agents regulating the core molecular pathway of breast cancer such as Wnt/ß-catenin signaling. METHODS: The present study screened a novel inhibitor, called "C188", using MTT assay. The molecular formula of C188 is C21H15FN4O3 and the molecular weight is 390. Flow cytometry and Western blotting were employed to assess cell cycle arrest after treatment with C188. Wound-healing and transwell assays were applied to measure the cell migration and invasion viability. The regulatory effects of C188 on Wnt/ß­catenin signaling and localization of ß­catenin in the nucleus were investigated by Western blotting and immunofluorescence. RESULTS: We found that C188 significantly suppressed proliferation and growth in a dose- and time-dependent manner in breast cancer cells, but not in normal breast cells. The inhibitory effect was caused by cell cycle arrest at the G1-phase which is induced by C188 treatment. Additionally, C188 dramatically inhibited cell migration of breast cancer cells in a dose-dependent manner. The migration inhibition was attributed to the suppression of Wnt/ß­catenin signaling and localization of ß­catenin in the nucleus mediated by regulating phosphorylation of ß­catenin and its subsequent stability. Furthermore, the target genes, including Axin 2, c-JUN, and c-Myc, were downregulated due to the decrease of ß­catenin in the nucleus after exposure to C188. CONCLUSION: C188 treatment resulted in the downregulation of cyclin D which led to cell cycle arrest at the G1 phase, and the inhibition of cell migration, indicating that C188 may be an effective novel therapeutic candidate as a potential treatment for human breast cancer.

A Novel Imidazopyridine Derivative Exerts Anticancer Activity by Inducing Mitochondrial Pathway-Mediated Apoptosis.

BACKGROUND: Cancer remains a major clinical challenge because of the lack of effective drug for its treatment. To find out novel cancer chemotherapeutic molecules, we explored the anticancer effect of novel imidazopyridine compound 9i and also investigated the underlying molecular mechanism. METHODS: Human cervical cancer cell (HeLa) viability was measured by an MTT assay after treatment with compound 9i. Clonogenicity of HeLa cells was investigated by an in vitro colony formation assay. Cell death was visualized by propidium iodide (PI) staining. Fluorescence-activated cell sorting (FACS) was used to determine apoptosis and mitochondrial membrane potential in HeLa cells. The expression level of apoptosis-related proteins was also determined by western blot. RESULTS: Compound 9i suppressed HeLa cell viability in a time- and dose-dependent manner. Compound 9i induced mitochondrial outer membrane permeabilization (MOMP), activated caspase cascade, and finally resulted in apoptosis. CONCLUSION: Compound 9i induces mitochondrial pathway-mediated apoptosis in human cervical cancer cells, suggesting that 9i could be a potential lead compound to be developed as a cancer therapeutic molecule.

One-pot synthesis of substituted pyrrole-imidazole derivatives with anticancer activity.

A facile and efficient method to synthesize pyrrole-imidazole was developed via a post-Ugi cascade reaction followed by one purification procedure. Synthesized pyrrole-imidazole was collected by performing a mild reaction and a simple procedure, which could be applicable to a broad scope of functionalized anilines. The screening results demonstrated that compound 7e exhibited a high potency of anticancer activity in human pancreatic cancer cell lines PANC and ASPC-1. Our work shed light on the potential of post-Ugi cascade reaction in combinatorial and medicinal chemistry.

Anticancer activity of globularifolin against human adenoid cystic carcinoma cells is due to ROS-mediated apoptotic cell death and modulation of the JAK/STAT signalling pathway.

PURPOSE: Adenoid cystic carcinoma is a rare and under-researched disease. There is hardly any chemotherapy available for it, hence the urgent need to develop novel and efficient chemotherapy. Therefore, we examined the anticancer effects of globularifolin, an acylated iridoid glucoside, against salivary adenoid cystic carcinoma (SACC-83) cell line and normal human salivary gland (HSG) cell line. METHODS: Cell counting and colony formation assays were used to determine cell viability. Acridin orange (AO)/ethidium bromide (EB) staining and comet assay were used for the detection of apoptosis. Reactive oxygen species (ROS) determination and cell cycle analysis were performed by flow cytometry. Transwell assay was used to monitor cell migration and Western blot analysis was used to determine protein expression. RESULTS: Globularifolin inhibited the growth of SACC-83 cell line and exhibited an IC50 of 10 µM. Nonetheless, the cytotoxic effects of globularifolin were comparatively negligible against normal HGS cells with an IC50 of 80 µM. The investigation of the mechanism of action revealed that the anticancer effects of globularifolin against the SACC-83 cells was due to the induction of apoptotic cell death as indicated by AO/EB staining. Globularifolin treatment also resulted in enhancement of the Bax, Caspase 3 and 9 expression and decline of the Bcl-2 expression. Globularifolin also blocked the SACC-83 cells at the G0/G1 phase of the cell cycle. Moreover, cell invasion assay revealed that globularifolin inhibited the migration of the SACC-83 cells concentration-dependently, which was also coupled with the downregulation of metalloproteinase (MMP) 2 and 9. JAK/STAT is an important pathway involved in the proliferation and tumorigenesis of cancer cells and this research found that globularifolin could inhibit this pathway. CONCLUSION: We conclude that globularifolin may prove essential in the development of systemic therapy for adenoid cystic carcinoma.

A Derivate of Benzimidazole-Isoquinolinone Induces SKP2 Transcriptional Inhibition to Exert Anti-Tumor Activity in Glioblastoma Cells.

We have previously shown that compound-7g inhibits colorectal cancer cell proliferation and survival by inducing cell cycle arrest and PI3K/AKT/mTOR pathway blockage. However, whether it has the ability to exert antitumor activity in other cancer cells and what is the exact molecular mechanism for its antiproliferation effect remained to be determined. In the present study, compound-7g exhibited strong activity in suppressing proliferation and growth of glioblastoma cells. The inhibitor selectively downregulated F-box protein SKP2 expression and upregulated cell cycle inhibitor p27, and then resulted in G1 cell cycle arrest. Mechanism analysis revealed that compound-7g also provokes the down-regulation of E2F-1, which acts as a transcriptional factor of SKP2. Further results indicated that compound-7g induced an increase of LC3B-II and p62, which causes a suppression of fusion between autophagosome and lysosome. Moreover, compound-7g mediated autophagic flux blockage promoted accumulation of ubiquitinated proteins and then led to endoplasmic reticulum stress. Our study thus demonstrated that pharmacological inactivation of E2F-1-SKP2-p27 axis is a promising target for restricting cancer progression.

Application of apically positioned flap technique on keratinized gingiva reconstruction in implant / 口腔疾病防治

Objective @#To explore the method of apically positioned flap technique (APFT) on buccal keratinized gingiva reconstruction around dental implants and evaluate its clinical outcomes and technical characters. @*Methods @#13 patients, who were insufficient of buccal keratinized gingiva around dental implants but sufficient with alveolar ridge crest tissue or palatal tissue at posterior maxillar, were enrolled in this study. APFT was operated during the phase Ⅱ surgery, by which some tissues were transferred from the alveolar ridge crest or palatal side to the original site of keratinized gingiva. Before APFT surgery and 1, 6 and 12 months after crown restoration, the width and thickness of transferred tissues were measured, gingival index (GI) and bleeding on probing (BOP) were also detected. The results were applied to comparative t-test statistical analysis.@*Results @# Transferred tissues by APFT showed healthy one month after crown restoration and exhibited characters of keratinized gingiva compared with the adjacent teeth at 6 and 12 months after restoration. Mean value of width of transferred tissue were respectively (3.25 ± 0.40) mm, (3.04 ± 0.34) mm and (2.97 ± 0.32) mm, meanwhile the thickness were respectively (2.05 ± 0.20) mm, (1.91 ± 0.23) mm and (1.84 ± 0.25) mm. The value of width and thickness of the adjacent teeth gingiva were (3.19 ± 0.42) mm and (1.96 ± 0.23) mm respectively. No significant differences were found between transferred tissue and adjacent teeth gingiva on width and thickness (P>0.05). Observation results of GI and positive rate of BOP of transferred tissue were also similar to which of gingiva of the adjacent teeth. @*Conclusion @#Technique of apically positioned flap is an effective measure on buccal keratinized gingiva reconstruction.

[Effects of Choukroun's platelet-rich fibrin on human gingival fibroblasts proliferation, migration and type I collagen secretion].

OBJECTIVE: To investigate the effect of platelet-rich fibrin (PRF) on human gingival fibroblasts (HGF) biological behavior such as proliferation, migration and collagen-I expression. METHODS: Human healthy gingival tissues were cultured according to the explant technique to obtain primary cultures. PRF was prepared by means of Choukroun's. HGF were co-cultured with PRF membrane originating from the same donor as the explants, divided into three groups, PRF1 group, PRF2 group and blank control group. Methyl thiazolyl tetrazolium (MTT) assay was used for cytotoxicity and cell proliferation study, and enzyme-linked immunosorbent assay (ELISA) for collagen-I (COL-I) secretion study at the 1st, 3rd, 5th day respectively. Eluates from PRF membrane was prepared, and divided into three groups, PRF1 group, PRF2 group and blank control group. Transwell chamber was utilized to determine the effect of PRF membrane eluate on cell migration. RESULTS: The A values of HGF in culture of the PRF1 (0.615 ± 0.036, 0.686 ± 0.006, 0.693 ± 0.004) and PRF2 groups (0.653 ± 0.023, 0.766 ± 0.034, 0.775 ± 0.053) were significantly higher than those of the control cultures (0.514 ± 0.020, 0.544 ± 0.006, 0.545 ± 0.009) (P < 0.01), but the difference between PRF1 and PRF2 group was not significant (P > 0.05). In each group at different time points, the HGF proliferation effect was significantly enhanced with time (P < 0.01). Cell migration test showed that the migration numbers of HGF in PRF1 and PRF2 groups (85.67 ± 2.94, 85.83 ± 1.47) were significantly higher than those of the control group (54.17 ± 2.48) (P < 0.01), but the difference between the two experimental groups was not significant (P > 0.05). COL-I secretion test exhibited that the A values of COL-I in PRF1 (0.184 ± 0.004, 0.200 ± 0.004, 0.204 ± 0.009) and PRF2 group (0.213 ± 0.008, 0.226 ± 0.005, 0.229 ± 0.006) were significantly higher than the A values of the control group (0.174 ± 0.002, 0.184 ± 0.002, 0.186 ± 0.003) (P < 0.01), but the difference between the two experimental groups was not significant (P > 0.05). In each group, the secretion level of COL-I increased significantly with time (P < 0.01). CONCLUSIONS: PRF could exert a positive effect on HGF biological behaviour and had clinical application potential in the treatment of gingival recession and in the periodontal tissue engineering when combined with seed cell HGF.

[Isolation and characterization of a DRE-binding transcription factor from Yinxin poplar (Populus alba x P. alba var. pyramidalis)].

Dehydration-Responsive Element Binding ( DREB) transcription factors, specifically binding with dehydration reponsive element (DRE), activate a variety of stress-responsive genes in plants under abiotic stresses (dehydration, high salt and low temperature). Using PCR and homologous EST search, we isolated a DREB-like gene from Yinxin poplar (Populus alba x P. alba var. pyramidalis) named PaDREB2. Yeast One-hybrid experiment demonstrated that PaDREB2 protein could function as a DREB transcription factor activating target gene expression by specifically binding to DRE cis-element. To study the expression pattern of PaDREB2, RT-PCR was carried out. And the results showed that PaDREB2 is induced by low temperature, drought and high salt.