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Isoquinoline alkaloids possess broad pharmacological activities. In this study, the antifungal activity of twelve isoquinoline alkaloids, including berberine (1), jatrorrhizine (2), coptisine (3), corydaline (4), tetrahydroberberine (5), chelidonine (6), dihydrosanguinarine (7), chelerythrine (8), sanguinarine (9), palmatine (10), tetrahydropalmatine (11) and columbamine (12) were evaluated against eight plant pathogenic fungi in vitro. All the tested compounds showed varying degrees of inhibition against the eight tested plant fungi. Among them, sanguinarine exhibited high antifungal activity (EC50 ranging from 6.96-59.36 μg/mL). It displayed the best inhibitory activity against Magnaporthe oryzae (EC50 = 6.96 μg/mL), compared with azoxystrobin (EC50 = 12.04 μg/mL), and significantly suppressed spore germination of M. oryzae with the inhibition rate reaching 100% (50 μg/mL). The optical microscopy and scanning electron microscopy observations revealed that after treating M. oryzae mycelia with sanguinarine at 10 μg/mL, the mycelia appeared curved, collapsed and the cell membrane integrity was eventually damaged. Furthermore, the reactive oxygen species production, mitochondrial membrane potential and nuclear morphometry of mycelia had been changed, and the membrane function and cell proliferation of mycelia were destroyed. These results will enrich our insights into action mechanisms of antifungal activity of sanguinarine against M. oryzae.
Fungicidal activity; Isoquinoline alkaloids; Sanguinarine.
Anti-phytopathogenic activity and the possible mechanisms of action of isoquinoline alkaloid sanguinarine
Zhong-Min Zhao 1, Xiao-Fei Shang 2, Raymond Kobla Lawoe 1, Ying-Qian Liu 3, Rui Zhou 1, Yu Sun 1, Yin-Fang Yan 1, Jun-Cai Li 1, Guan-Zhou Yang 1, Cheng-Jie Yang 1
Rationale: Levo-tetrahydropalmatine (l-THP), an active ingredient of Corydalis yanhusuo, has been reported to be a partial agonist for dopamine D1 receptors (D1R) and an antagonist for D2R. Although it has been safely used clinically in China for decades as an analgesic with sedative/hypnotic properties, there are few studies that address the mechanisms by which l-THP exerts its beneficial effects in chronic pain-induced sleep disturbance.
Objectives: To investigate the effects and mechanisms of l-THP on sleep disturbance in a neuropathic pain-like condition.
Methods: A mouse model of chronic neuropathic pain induced by partial sciatic nerve ligation (PSNL) was employed. The antinociceptive and hypnotic effects of l-THP were evaluated by measurement of mechanical allodynia, thermal hyperalgesia, and electroencephalogram (EEG) recordings in PSNL mice. Pharmacological approaches and c-Fos expression were used to clarify the mechanisms of l-THP.
Results: Intraperitoneal injection of l-THP at 5 and 10 mg/kg not only significantly increased the mechanical threshold by 134.4% and 174.8%, and prolonged the thermal latency by 49.4% and 69.2%, but also increased non-rapid eye movement sleep by 17.5% and 29.6%, and decreased sleep fragmentation in PSNL mice, compared with the vehicle control. Moreover, the antinociceptive effect of l-THP was prevented by D1R antagonist SCH23390 or D2R agonist quinpirole; meanwhile, the hypnotic effect of l-THP was blocked by quinpirole rather than by SCH23390. Immunohistochemistry demonstrated that l-THP inhibited c-Fos overexpression induced by PSNL in the cingulate cortex and the periaqueductal gray.
Conclusions: These findings indicated that l-THP exerted analgesic effects by agonism D1R and antagonism D2R, and the antagonism of D2R mediated the hypnotic effect of l-THP in PSNL mice.
Dopamine receptor; Levo-tetrahydropalmatine; Neuropathic pain; Sleep disturbance; c-Fos.
Dopamine D 1 and D 2 receptors mediate analgesic and hypnotic effects of l-tetrahydropalmatine in a mouse neuropathic pain model
Yuan-Yuan Liu 1, Tian-Xiao Wang 1, Ji-Chuan Zhou 1, Wei-Min Qu 2, Zhi-Li Huang 3
Purpose: Το evaluate the effect of L-Tetrahydropalmatine (L-THP) on the sensitivity of a cisplatin resistant ovarian cancer (OC) cell line. As miR-93 is reported to be overexpressed in OC and cisplatin resistance, we also evaluated its pathway in OC.
Methods: The levels of miR-93 were evaluated using RT-PCR and Luciferase assay was performed to confirm the target of miR-93. The extent of apoptosis was evaluated by Annexin V and propidium iodide (PI) staining, whereas Hoechst 33258 staining was done for identifying the number of apoptotic cells.
Results: The cisplatin-resistant A2780/DDP cell line showed lower survival rate compared to control when incubated with L-THP along with cisplatin. L-THP caused G0/G1 cell cycle arrest and increased the sensitivity to cisplatin. Furthermore, we found that the levels of miR-93 in cisplatin-resistant cells were highly expressed compared to parental cells. L-THP suppressed the expression of miR-93 and increased the levels of PTEN, a crucial tumor suppressor in OC. It was further observed that the cells transfected with PTEN siRNA showed increased survival compared with the control group and this phenomenon could be reversed by the AKT inhibitor Triciribine. The A2780 cells treated with PTEN siRNA showed similar survival rate to the cells with miR-93 overexpression.
Conclusion: The findings of this study suggested L-THP increased the sensitivity of ovarian cancer cells to cisplatin via modulating miR-93/PTEN/AKT pathway in A2780/DDP ovarian cancer cell line.
L-Tetrahydropalmatine enhances the sensitivity of human ovarian cancer cells to cisplatin via microRNA-93/PTEN/Akt cascade
Jian Gong 1, Chao Xing, Lian Yun Wang, Shuang Shuang Xie, Wen Dong Xiong