Quantitative HPLC analysis of benzene derivatives of Melicope ptelefolia leaves

A high performance liquid chromatography procedure for the quantitative determination of three marker benzene derivatives, 2,4,6-trihydroxy-3-prenyl acetophenone (tHPA) (1), 2,4,6-trihydroxy-3-geranyl acetophenone (tHGA) (2), and p-O-geranyl coumaric acid (GCA) (3), in the Melicope ptelefolia ethanolic leaf extracts, a medicinal herb obtained from a few locations of the Peninsula Malaysia, was described. The quantitative analysis was performed using high performance liquid chromatography-photodiode array detection on Xterra octadecylsiyl silica (ODS; 3.0 × 150 mm, 3.5 μm) column kept at 32°C, using gradient elution with acetonitrile and water containing 0.1% formic acid at a flow-rate of 1 ml/min with UV detection wavelength at 280 nm. All calibration curves showed good linearity (R2 of 0.999 to 1.0000) within the concentrations range of 2.5 × 10-3 to 0.1 mg/mL. The method was shown to be simple, sensitive, and reliable for qualitative and quantitative analysis of the marker compounds in M. ptelefolia leaf preparations

Observation on SPME different headspace fiber coupled with GC-MS in extracting high quality agarwood chipwood

Agarwood is well known as one of the expensive woods in the world. It has a unique scent which brings it to have wide usages especially in perfumery ingredient, as incense, in traditional medical preparation, and as symbol of wealth. Due to that, this paper presents the analysis on chemical profiles of agarwood chipwood, as a part of agarwood grading system. The work involved of Solid Phase Microextraction (SPME) coupled with Gas Chromatography - Mass Spectrometry (GC-MS) GC-MS in extracting high quality. Three headspace fibers; PDMS-DVB, CAR-PDMS and DVB-CAR-PDMS were used during the extraction to identify the compounds with the sampling time of 60 minutes. The result showed that high quality agarwood chipwood is made up of terpene group which are monoterpene hydrocarbon, sesquiterpene hydrocarbon and oxygenated sesquiterpene. The relative peak areas (%) for compounds are tabulated and plotted. The finding in this study confirmed that the difference in compounds extracted and their relative peak area (%) are due to different fiber's polarity and absorbent, Thus, it is significant and benefit especially in agarwood oil quality grading and its related area

Gas chromatography mass spectrometry couple with quadrupole time-of-flight (GC-QTOF MS) as a powerful tool for profiling of oxygenated sesquiterpenes in agarwood oil

Agarwood (Aquilaria malaccensis) is very well known as the most expensive wood in the world due to its wide applications in perfumery, cosmetic traditional medicine, and religious ceremonies. The study aimed to give an in-depth characterisation focusing on marker compounds in A. malaccensis from different places in Malaysia. The establishment of an oxygenated sesquiterpenes chemical profile of the fungus-infected agarwood oil was achieved by gas chromatography mass spectrometry (GC–MS) coupled with quadrupole time (QTOF) technique. Aroma compounds were identified as sesquiterpenes and oxygenated sesquiterpenes where agarospirol was found in samples of all locations (3.12%, 3.54%, 3.36% and 2.26% from Melaka, Pahang, Kelantan A and Kelantan B respectively) and also N-hexadecanoic acid as one of the major compounds. Both compounds were further isolated by Prep-GC and confirmed by NMR. This study provides a reference for agarwood oil analysis from different origins in Malaysia

Discrimination of young and mature leaves of Melicope ptelefolia using 1H NMR and multivariate data analysis

The ‘Ulam’, a traditional Malay dish, are plants that can be eaten raw, as a form of local salad. The shoots and young leaves of Melicope ptelefolia are among the popular species, believed to be high in nutritional and medicinal values. The metabolomic fingerprinting analysis of the ethanolic extracts of leaves of M. ptelefolia was carried out using 1H Nuclear Magnetic Resonance (NMR) spectroscopy and multivariate data analysis in order to differentiate young and mature leaves and to evaluate the variation of their chemical composition. Principle component analysis (PCA) of the 1H NMR spectra showed a clear discrimination between the young and mature leaves extracts by PC3 and PC4. The compounds responsible for the differentiation were identified by comparison of 1H NMR chemical shifts and qualitative HPLC. The young leaves were found to be richer in fatty acids and the levels of the three marker compounds, p-O-geranylcoumaric acid, 2,4,6-trihydroxy-3-geranylacetophenone and 2,4,6-trihydroxy 3-prenylacetophenone, were clearly higher. The mature leaves contain higher levels of sugars and glycosidic components

Synthesis and cytotoxic effects of (E)-3-(2,3-dimethoxyphenyl)-1-(5-methylfuran-2-yl) prop-2-en-1-one in MDA-MB231 and MCF-7 breast cancer cell lines

A chalcone derivative, (E)-3-(2,3-dimethoxyphenyl)-1-(5-methylfuran-2-yl)-prop-2-en-1-one (DMMF) was synthesized and evaluated against various cancerous cell lines including colon adenocarcinoma (HT-29), myloplasticleukemia (HL60), breast cancer (MCF-7 and MDA-MB231), normal hepatic cell (WRL-68) and normal breast cell (MCF-10A). The structure of DMMF was determined by EI-MS, 1H NMR and single X-ray crystallographic techniques. The DMMF possessed the highest cytotoxic effect against MCF-7 breast cancer cell (2.01 ± 1.53 μg/mL) and lowest against normal hepatic WRL-68 and breast cell lines after 24 h of treatment. Induction of apoptosis and regulation of cell cycle progression results indicates the significant increase in early apoptosis and G2/M arrest after 48 h of treatment in MCF-7 cells. Meanwhile, in MDA-MB231 cells, there was an increase in Sub G0/G1 cells population and early/late apoptotic cells upon treatment with DMMF. Additionally, DMMF effectively induced G2/M cell cycle arrest in MCF-7 cells and apoptosis in both MCF-7 and MDA-MB231 cells

Bioassay-guided identification of an anti-inflammatory prenylated acylphloroglucinol from Melicope ptelefolia and molecular insights into its interaction with 5-lipoxygenase.

A bioassay-guided investigation of Melicope ptelefolia Champ ex Benth (Rutaceae) resulted in the identification of an acyphloroglucinol, 2,4,6-trihydroxy-3-geranylacetophenone or tHGA, as the active principle inhibiting soybean 15-LOX. The anti-inflammatory action was also demonstrated on human leukocytes, where the compound showed prominent inhibitory activity against human PBML 5-LOX, with an IC 50 value of 0.42 μM, very close to the effect produced by the commonly used standard, NDGA. The compound concentration-dependently inhibited 5-LOX product synthesis, specifically inhibiting cysteinyl leukotriene LTC4 with an IC 50 value of 1.80 μM, and showed no cell toxicity effects. The anti-inflammatory action does not seem to proceed via redox or metal chelating mechanism since the compound tested negative for these bioactivities. Further tests on cyclooxygenases indicated that the compound acts via a dual LOX/COX inhibitory mechanism, with greater selectivity for 5-LOX and COX-2 (IC 50 value of 0.40 μM). The molecular features that govern the 5-LOX inhibitory activity was thus explored using in silico docking experiments. The residues Ile 553 and Hie 252 were the most important residues in the interaction, each contributing significant energy values of 13.45 (electrostatic) and 5.40 kcal/mol (electrostatic and Van der Waals), respectively. The hydroxyl group of the phloroglucinol core of the compound forms a 2.56 Å hydrogen bond with the side chain of the carboxylate group of Ile 553. Both Ile 553 and Hie 252 are crucial amino acid residues which chelate with the metal ion in the active site. Distorting the geometry of these ligands could be the reason for the inhibition activity shown by tHGA. The molecular simulation studies supported the bioassay results and served as a good model for understanding the way tHGA binds in the active site of human 5-LOX enzyme

Design, synthesis and cytotoxic effects of curcuminoids on HeLa, K562, MCF-7 and MDA-MB-231 cancer cell lines

Background Curcumin is one of the leading compound extracted from the dry powder of Curcuma longa (Zingiberaceae family), which possess several pharmacological properties. However, in vivo administration exhibited limited applications in cancer therapies. Results Twenty-four curcumin derivatives have synthesized, which comprises cyclohexanone 1–10, acetone 11–17 and cyclopentanone 18–24 series. All the curcuminoids were synthesized by the acid or base catalyzed Claisen Schmidt condenstion reactions, in which β-diketone moiety of curcumin was modified with mono-ketone. These curcuminoids 1–24 were screened against HeLa, K562, MCF-7 (an estrogen-dependent) and MDA-MB-231 (an estrogen-independent) cancer cell lines. Among them, acetone series 11–17 were found to be more selective and potential cytotoxic agents. The compound 14 was exhibited (IC50 = 3.02 ± 1.20 and 1.52 ± 0.60 µg/mL) against MCF-7 and MDA-MB-231 breast cancer cell lines. Among the cyclohexanone series, the compound 4 exhibited (IC50 = 11.04 ± 2.80, 6.50 ± 01.80, 8.70 ± 3.10 and 2.30 ± 1.60 µg/mL) potential cytotoxicity against four proposed cancer cell lines, respectively. All the curcucminoids were characterized with the detailed 1H NMR, IR, UV–Vis, and mass spectroscopic techniques. The structure of compound 4 was confirmed by using the single X-ray crystallography. Additionally, we are going to report the first time spectral data of (2E,6E)-2,6-bis(2-methoxybenzylidene)cyclohexanone (1). Structure–activity relationships revealed that the mono-carbonyl with 2,5-dimethoxy substituted curcuminoids could be an essential for the future drugs against cancer diseases. Conclusions Curcuminoids with diferuloyl(4-hydroxy-3-methoxycinnamoyl) moiety with mono carbonyl exhibiting potential cytotoxic properties. The compound 14 was exhibited (IC50 = 3.02 ± 1.20 and 1.52 ± 0.60 µg/mL) against MCF-7 and MDA-MB-231 breast cancer cell lines

Possible participation of nitric oxide/cyclic guanosine monophosphate/protein kinase C/ATP-sensitive K + channels pathway in the systemic antinociception of flavokawin B.

The possible mechanisms of action in the antinociceptive activity induced by systemic administration (intraperitoneal, i.p.) of flavokawin B (FKB) were analysed using chemical models of nociception in mice. It was demonstrated that i.p. administration of FKB to the mice at 0.3, 1.0, 3.0 and 10 mg/kg produced significant dose-related reduction in the number of abdominal constrictions. The antinociception induced by FKB in the acetic acid test was significantly attenuated by i.p. pre-treatment of mice with l-arginine, the substrate for nitric oxide synthase or glibenclamide, the ATP-sensitive K+ channel inhibitor, but was enhanced by methylene blue, the non-specific guanylyl cyclase inhibitor. FKB also produced dose-dependent inhibition of licking response caused by intraplantar injection of phorbol 12-myristate 13-acetate, a protein kinase C activator (PKC). Together, these data indicate that the NO/cyclic guanosine monophosphate/PKC/ATP-sensitive K+ channel pathway possibly participated in the antinociceptive action induced by FKB

Induction of apoptosis and regulation ofMicroRNA expression by (2E,6E)-2,6-bis-(4-hydroxy-3-methoxybenzylidene)- cyclohexanone (BHMC) treatment on MCF-7 breast cancer cells

(2E,6E)-2,6-bis-(4-hydroxy-3-methoxybenzylidene)-cyclohexanone (BHMC) is a synthetic curcumin analogue, which has been reported to possess anti-tumor, anti-metastatic, and anti-invasion properties on estrogen receptor (ER) negative breast cancer cells in vitro and in vivo. However, the cytotoxic effects of BHMC on ER positive breast cancer cells were not widely reported. This study was aimed to investigate the cytotoxic potential of BHMC on MCF-7 cells using cell viability, cell cycle, and apoptotic assays. Besides, microarray and quantitative polymerase chain reaction (qPCR) were performed to identify the list of miRNAs and genes, which could be dysregulated following BHMC treatment. The current study discovered that BHMC exhibits selective cytotoxic effects on ER positive MCF-7 cells as compared to ER negative MDA-MB-231 cells and normal breast cells, MCF-10A. BHMC was shown to promote G2/M cell cycle arrest and apoptosis in MCF-7 cells. Microarray and qPCR analysis demonstrated that BHMC treatment would upregulate several miRNAs like miR-3195 and miR-30a-3p and downregulate miRNAs such as miR-6813-5p and miR-6132 in MCF-7 cells. Besides, BHMC administration was also found to downregulate few tumor-promoting genes like VEGF and SNAIL in MCF-7. In conclusion, BHMC induced apoptosis in the MCF-7 cells by altering the expressions of apoptotic-regulating miRNAs and associated genes

Effects of Payena dasyphylla (Miq.) on hyaluronidase enzyme activity and metalloproteinases protein expressions in interleukin-1beta stimulated human chondrocytes cells

Background: Hyaluronidases have been found as the target enzymes in the development of osteoarthritis (OA) disease. While there is still no curative treatment for this disease, recent studies on the treatment of OA were focused on the effectiveness of natural products which are expected to improve the symptoms with minimal side effects. The aim of this study was to screen selected Malaysian plants on their anti-hyaluronidase activity as well as to evaluate the active plant and its derived fractions on its potential anti-arthritic and antioxidant activities.Methods: A total of 20 methanolic crude extracts (bark and leaf) from ten different plants were screened using a colorimetric hyaluronidase enzymatic assay. The active plant extract (Payena dasyphylla) was then studied for its hyaluronidase inhibitory activity in the interleukin-1β (IL-1β) stimulated human chondrocytes cell line (NHAC-kn) using zymography method. The Payena dasyphylla methanolic bark extract was then fractionated into several fractions in where the ethyl acetate (EA) fraction was evaluated for its inhibitory effects on the HYAL1 and HYAL2 gene expressions using reverse transcription-polymerase chain reaction (RT-PCR) technique. While the MMP-3 and MMP-13 protein expressions were evaluated using western blot method. The phenolic and flavonoid contents of the three fractions as well as the antioxidant property of the EA fraction were also evaluated.Results: Bark extract of Payena dasyphylla (100 μg/ml) showed the highest inhibitory activity against bovine testicular hyaluronidase with 91.63%. The plant extract also inhibited hyaluronidase expression in the cultured human chondrocyte cells in response to IL-1β (100 ng/ml). Similarly, treatment with Payena dasyphylla ethyl acetate (EA) fraction (100 μg/ml) inhibited the HYAL1 and HYAL2 mRNA gene expressions as well as MMP-3 and MMP-13 protein expression in a dose dependent manner. Payena dasyphylla EA fraction has demonstrated the highest amount of phenolic and flavonoid content with 168.62 ± 10.93 mg GAE/g and 95.96 ± 2.96 mg RE/g respectively as compared to water and hexane fractions. In addition, the Payena dasyphylla EA fraction showed strong antioxidant activity with IC50 value of 11.64 ± 1.69 μg/mL.Conclusion: These findings have shown that Payena dasyphylla might contained potential phenolic compounds that inhibiting the key enzyme in osteoarthritis development, which is the hyaluronidase enzyme through interruption of HYAL1 and HYAL1 gene expressions. The degradation of cartilage could also be inhibited by the plant through suppression of MMP-3 and MMP-13 protein expressions. We also reported that the inhibitory effect of Payena dasyphylla on hyaluronidase activity and expression might be due to its anti-oxidant property