Protective effects of 3′‐deoxy‐4‐O‐methylepisappanol from Caesalpinia sappan against glutamate‐induced neurotoxicity in primary cultured rat cortical cells

To examine the neuroprotective effects of Caesalpinia sappan L., we tested its protection against the glutamate‐induced neurotoxicity in primary cortical cultured neurons. We found that an aqueous extract of this medicinal plant exhibited significant protection against glutamate‐induced toxicity in primary cultured rat cortical cells. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. By such fractionation, two known compounds – sappanchalcone and 3′‐deoxy‐4‐O‐methylepisappanol – were isolated from the methanol extracts from the air‐dried and chipped C. sappan. Among these two compounds, 3′‐deoxy‐4‐O‐methylepisappanol exhibited significant neuroprotective activities against glutamate‐induced toxicity, exhibiting cell viability of about 50%, at concentrations ranging from 0.1 μM to 10 μM. Therefore, the neuroprotective effect of C. sappan might be due to the inhibition of glutamate‐induced toxicity by the protosappanin derivative it contains. Copyright © 2009 John Wiley & Sons, Ltd.     

3种国产云实属植物的单宁酸及水解产物的研究

目的:在国产云实属植物中寻找能替代进口塔拉生产单宁酸及没食子酸的品种资源。方法:用水浸法提取单宁酸,用碱水解法对单宁浸提浓缩液进行水解,对水解产物进行了鉴别,并进行了实验室规模试验。结果:3种植物单宁酸的含量(干基)分别为32.2%,46.6%,58.5%,折合原料与水解产物之比为:154∶1,11.8∶1,3.09～2.79∶1。结论:首次在我国云实属植物中找到了可替代进口塔拉的植物。     

A new cassane diterpenoid from the seeds of Caesalpinia decapetala

Phytochemical investigation on the seeds of Caesalpinia decapetala led to the isolation of a new cassane diterpenoid with an unusual O bridge between C-19 and C-20, named phanginin Q (1), together with three known cassane diterpenoids, caesaljapin (2), caesaldekarin A (3), and caesaldekarin B (4). The structure of the new compound was elucidated by spectroscopic methods, including ¹H NMR, ¹³C NMR, HSQC, ¹H ? ¹H COSY, HMBC, NOESY, and HR-ESI-MS.     

云实种子中1个新颖卡山烷二萜类化合物

目的研究云实Caesalpinia decapetala种子的化学成分。方法采用硅胶、Sephadex LH-20、ODS等柱色谱技术进行分离纯化,采用各种波谱技术进行结构鉴定。结果从云实种子95%甲醇提取物中分离得到了1个新卡山烷二萜类化合物,鉴定为1α,2α-diacetoxy-14β-carboxymethyl-7β-hydro xyl-vouacapen-5α-ol(1)。结论化合物1为新的卡山烷二萜类化合物,命名为云实子C。     

In vitro Antitubercular Activity of 3‐Deoxysappanchalcone Isolated From the Heartwood of Caesalpinia sappan Linn.

Responsible for nearly 1.5 million deaths every year, the infectious disease tuberculosis remains one of the most serious challenges to global health. The emergence of multidrug‐resistant tuberculosis and, more recently, extensively drug‐resistant tuberculosis poses a significant threat in our effort to control this epidemic. New drugs are urgently needed to combat the growing threat of antimicrobial resistance. To achieve this goal, we screened approximately 500 species of medicinal plant methanol extracts and their solvent partitioned fractions for potential inhibitors of Mycobacterium tuberculosis growth. Using microdilution screening, the ethyl acetate solvent partitioned fraction from the heartwood of Caesalpinia sappan exhibited strong antitubercular activity. We isolated the active compound and identified it as 3‐deoxysappanchalcone. The extracted 3‐deoxysappanchalcone possessed activity against both drug‐susceptible and drug‐resistant strains of M. tuberculosis at MIC₅₀s of 3.125–12.5 μg/mL in culture broth and MIC₅₀s of 6.25–12.5 μg/mL inside macrophages and pneumocytes. 3‐Deoxysappanchalcone was also found to act in partial synergy with streptomycin/ethambutol against M. tuberculosis H37Rv. 3‐Deoxysappanchalcone had no cytotoxicity against the A549 cell line up to a concentration of 100 μg/mL (selectivity index > 8–32). Further studies are warranted to establish the in vivo effect and therapeutic potential of 3‐deoxysappanchalcone. Copyright © 2017 John Wiley & Sons, Ltd.     

Synergistic Cytotoxic and Antimigratory Effect of Brazilein and Doxorubicin on HER2-Overexpressing Cells

Objective: The present research aims to report cytotoxic and antimigratory activities of the oxidized form of brazilin, i.e., brazilein, and the effects of the combination of brazilein-doxorubicin on MCF-7/HER2 cells. Methods: The MTT assay was conducted to test the cytotoxic activity, while flow cytometry with PI and PI-annexin V staining were respectively performed for cell cycle and apoptosis analyses. Migration and invasion analyses were assessed via Boyden chamber assay, while HER2, Rac1, p120, MMP2, and MMP9 protein levels were determined by immunoblotting and gelatin zymography. Molecular docking of ligands with HER2, Src, PI3Kα, PI3KΔ, and PI3Kγ proteins was evaluated using MOE 2010. Results: The MTT assay showed that the IC50 value of brazilein against MCF-7/HER2 cells was 51 ± 2.1 µM. Moreover, brazilein and its combination with doxorubicin-induced G2/M accumulation and apoptosis. Combination of brazilein-doxorubicin inhibited cell migration and tended to decrease HER2, Rac1, p120, MMP2, and MMP9 protein expression levels. Based on our molecular docking study, the docking score of brazilein with PI3Kγ is comparable to that of the native ligand. Conclusion: Taken together, a combination of brazilein-doxorubicin exhibited synergistic cytotoxic and antimigratory effects on MCF-7/HER2 cells.     

Identification of active compounds from Caesalpinia sappan L. extracts suppressing IL-6 production in RAW 264.7 cells by PLS

Ethnopharmacological relevance

Caesalpinia sappan L. is distributed in Southeast Asia and also used as herbal medicine for the treatment of various diseases such as burning sensations, leprosy, dysentery, osteoarthritis and rheumatoid arthritis (RA). The overproduction of IL-6 plays an important role in the prognosis of RA, but the active compounds from the extracts of Caesalpinia sappan L. suppressing IL-6 production remain unknown.

Aims of the study

Identifying the main active compounds of Caesalpinia sappan L. extracts inhibiting the IL-6 production in LPS-stimulated RAW 264.7 cells by partial least squares (PLS).

Materials and methods

Sixty-four samples with different proportions of compounds were prepared from Caesalpinia sappan L. by supercritical CO₂ fluid extraction (SCFE) and refluxing. Each of 64 samples was applied to RAW 264.7 cells with LPS to evaluate whether IL-6 production by LPS is affected by addition of each sample. The IL-6 production in medium was determined by ELISA and the inhibitory activity of each sample was analyzed. In addition, the fingerprints of these 64 samples were also established by ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC–MS). We used the PLS, a simplified method, to evaluate the results from IL-6 production and fingerprints.

Results

Each of 64 samples markedly suppressed LPS-induced IL-6 production in RAW cells. The fingerprints by UPLC–MS clearly revealed variations among 64 samples produced in different extract conditions. The PLS analysis with IL-6 production and fingerprints by UPLC–MS suggested that the peaks 71, 93, 150, 157, 168 have more influence on the inhibitory activity of Caesalpinia sappan L. extracts. The peaks 71, 93, 150 are likely representing sappanone A, protosappanin E and neoprotosappanin, respectively. The peaks 157 and 168 are still at large.

Conclusion

This is the first report that sappanone A, protosappanin E, neoprotosappanin and two unidentified compounds can be considered as possible active compounds that might inhibit IL-6 production. Further studies are needed to confirm the effectiveness of these five compounds on IL-6 production and possible mechanism.     

苦石莲的化学成分研究

目的:研究苦石莲的化学成分。方法:利用硅胶柱色谱、重结晶等技术对苦石莲进行分离、纯化,通过波谱数据对化合物进行结构解析。结果:从苦石莲中分离纯化得到16个化合物,分别鉴定为caffeine(1)、β-谷甾醇(2)、十八烷酸(3)、熊果酸(4)、齐墩果酸(5)、β-胡萝卜苷(6)、3,5,7-三羟基-4’-甲氧基二氢黄酮醇(7)、neocaesalpin L(8)、neocaesalpin L1(9)、neocaesalpin K(10)、minaxin C(11)、阿魏酸(12)、咖啡酸(13)、没食子酸(14)、12α-ethoxyl-1α,6α,7β-triacetoxy-5α,14β-dihydroxy-cass-13(15)-en-16,12-olide(15)、stigmasta-5,22(E)-dien-3-beta-ol(16)。结论:化合物5,6,7,12,13,16均首次从该植物中分离得到,化合物7首次从豆科植物中分离得到。     

Brazilin Isolated from Caesalpina Sappan Wood Induces Intrinsic Apoptosis on A549 Cancer Cell Line by Increasing p53, caspase-9, and caspase-3

Objective: Lung cancer is the leading cause of death among cancer patients. The majority of lung cancer is the Non-Small Lung Carcinoma (NSLC). This study evaluated the potency of brazilin isolated from Caesalpinia sappan wood to induce apoptosis on non-small lung carcinoma cell line, A549, by examining the expression of p53, caspase-9, and caspase-3. Methods: Brazilin was isolated from Caesalpinia sappan wood following a guided assay and it was determined by using Brazilin®SIGMA as standard. The activity of brazilin on the growth of A549 cell line was analysed by MTT assay and the apoptosis was evaluated by flowcytometer following Annexin V (FITC) and PI staining. The expression of p53, caspase-9, and caspase-3 was examined by immunocytochemistry. Result: The IC50 of brazilin on A549 cell line was 43µg/mL. Cell treatment with 20 µg/mL and 40 µg/mL of brazilin significantly increased early apoptosis (p<0.001). Cell treatment with 40 µg/mL  of Brazilin significantly increased late apoptosis (p<0.001). Brazilin significantly increased the expression of p53, Caspase-9, and caspase-3 (p<0.001). Conclusion: This study showed evidence of the activity of brazilin to induce intrinsic apoptosis on a NSLC cell line A549.