Morushalunin D, a tri-O-bridged Diels-Alder type adduct from Morus alba var. shalun root cultures and the related precursors.

Three Diels-Alder type adducts (1-3) along with their precursors, including one 2-arylbenzofuran (4) and one stilbene (5), were isolated from the MeOH extract of M. alba var. shalun root cultures. Among them, 1 is a new Diels-Alder type adduct named morushalunin D. The molecular structures of 1-5 were elucidated based on spectroscopic data and comparison with the literatures. Cytotoxic properties of compounds 1-5 were evaluated against murine leukemia P-388 cells. Morushalunin D (1), mulberrofuran T (2), sorocein A (3), moracin M (4), and oxyresveratrol (5) were active, significantly inhibiting the growth of P-388 cells with IC50 values of 0.5, 1.0, 0.6, 2.0, and 3.3 µg/ml, respectively.

Xanthorrhizol derivatives and their biological properties as caspase-7 inhibitors.

Xanthorrhizol (1) is known as the major terpenoid component of the rhizome of Curcuma xanthorrhiza and having some interesting biological activities. In this report, we synthesised five derivatives of 1 containing nitrogen-functional groups. Four of them are new synthesised compounds, including (R)-4-(3-(2-methyl-5-(6-methylhept-5-en-2-yl)phenoxy)propyl)morpholine (2), (R)-N-benzyl-3-(2-methyl-5-(6-methylhept-5-en-2-yl)phenoxy)propan-1-amine (3), (R)-6,7-dimethoxy-3-(3-(2-methyl-5-(6-methylhept-5-en-2-yl)phenoxy)propyl)quinazolin-4(3H)-one (4), and (R)-6-methyl-3-(6-methylhept-5-en-2-yl)-2-nitrophenol (5) groups. Meanwhile the other is the known compound, that is (R)-2-methyl-5-(6-methylhept-5-en-2-yl)-4-nitrophenol (6). The caspase-7 inhibitory activity of compounds 1-6 was evaluated as well. In comparison to other derivatives, compounds 5 and 6 exhibited higher activity. Consequently, compounds 5 and 6 may be a promising lead compound for further development as a caspase-7 inhibitor.

Secondary metabolites from Colletotrichum gloeosporioides isolated from Artocarpus heterophyllus and evaluation of their cytotoxic and antibacterial activities.

A new lactone, collectindolide (1), as well as three known compounds, (S)-sydonic acid (2), (S)-hidroxysydonic acid (3), and indole-3-aceticacid (4) were obtained from the liquid media of Colletotrichum gloeosporioides, which was obtained from Artocarpus heterophyllus. The chemical structures of 1-4 were established by spectroscopic analyses, including NMR experiments and by HR-ESI-TOF-MS mass spectroscopy. Compounds 1-4 were evaluated for their cytotoxicity against murine leukaemia P-388 cell lines by MTT assay. Antibacterial activity of compounds 1-4 was also assayed against four bacteria. Phytochemical investigation of the genus Colletotrichum derived from the plant genus Artocarpus is reported for the first time.

Metabolite Profiles of the Green Beans of Indonesian Arabica Coffee Varieties.

The green beans of 3 Indonesian arabica coffee varieties, namely, ateng, buhun, and sigararutang, were analyzed with 1H NMR-based metabolomics coupled with alpha-glucosidase inhibitory activity assay. These coffees were cultivated in the same geographical conditions. The PLSDA model successfully classified the green coffee beans based on their varieties. To reveal the characteristic metabolites for each coffee variety, S-plot of two-class OPLSDA models was generated and analyzed. Ateng coffee was characterized with trigonelline, sucrose, 5-CQA, and acetic acid. The characteristic metabolites of buhun coffee were citric acid and malic acid. Meanwhile, the most discriminant compound of sigararutang coffee was quinic acid. HCA analysis revealed the lineage relationship of the 3 coffee varieties. Ateng coffee had closer lineage relationship to sigararutang compared to the buhun coffee. Alpha-glucosidase inhibitory activity of the coffee samples did not differ widely. IC50 values of alpha-glucosidase inhibitory activity of ateng, sigararutang, and buhun coffees were 3.01 ± 0.16, 3.14 ± 0.20, and 5.05 ± 0.28 mg/mL, respectively. Although grown in the same geographical conditions, our results revealed that each coffee variety possessed a unique metabolome clarifying the diversity of Indonesian arabica coffees. This study verified that 1H NMR-based metabolomics is an excellence method for discovering the lineage relationship in the samples with different varieties or cultivars.

Metabolite profiles and antidiabetic activity of the green beans of Luwak (civet) coffees.

Metabolite profiles of green beans (the caged and the wild) Luwak (civet) coffees were evaluated by NMR techniques combined with chemometrics. The bioactivities of the green coffee beans were examined with antioxidant tests and an alpha-glucosidase inhibitory assay. Both are invitro tests related to the antidiabetic properties. Our results showed the civet coffees possessed unique metabolomes and were different from the regular arabica coffee. Both civet coffees were characterized by higher concentrations of alanine, citrate, lactate, malate, and trigonelline. Lactate and lipids were found as the most important compounds discriminating the caged civet coffee from the wild civet coffee. Bioactivity assays exhibited the antidiabetic activities of the civet coffees were better than the activity of the regular coffee. These results suggested that the civet coffees are promising functional foods reducing the diabetes risk. It is the first report evaluating metabolite profiles of both civet coffees using 1H NMR-based metabolomics.

Potential cytotoxic Diels-Alder type adducts from liquid medium of Morus Alba var. shalun root cultures.

A new Diels-Alder type adduct named morushalunin (1), together with three similar compounds, guangsangon E (2), chalcomoracin (3), and kuwanon J (4) were isolated from the EtOAc extract of liquid medium of Morus alba var. shalun root cultures. The structures of the isolated compounds were determined based on spectroscopic data, including 1D, 2D NMR, and MS data. Cytotoxic properties of compounds 1-4 were evaluated against murine leukemia P-388 cells. Morushalunin (1), guangsangon E (2), and chalcomoracin (3) were found to have significant cytotoxicity with IC50 values 0.7, 2.5, and 1.7 µg/mL, respectively, while kuwanon J (4) showed weak cytotoxicity (IC50 = 5.9 µg/mL).

Secondary metabolites from Diaporthe lithocarpus isolated from Artocarpus heterophyllus.

Fractionation and purification of the ethyl acetate extract of Diaporthe lithocarpus, an endophytic fungus from the leaves of Artocarpus heterophyllus, yielded one new compound, diaporthindoic acid (1), along with seven known compounds (2-8). The new compound was characterized and established by the basis of extensive spectroscopic methods including NMR (1D and 2D) and HRMS. Compound 6 showed the best citotoxicity against murine leukemia P-388 cells with an IC50 value of 0.41 µg/mL. All compounds (1-8) were also tested for their antimicrobial activities. To the best of our knowledge, this is the first chemical evaluation of fungal Diaporthe derived from Artocarpus.

Sesquiterpenes produced by Pestalotiopsis microspora HF 12440 isolated from Artocarpus heterophyllus.

A drimane-type sesquiterpene, (+)-dendocarbin L (1) together with two bisabolane-type sesquiterpenes, (+)-sydonic acid (2) and (+)-sydowic acid (3) were isolated from the mycelium of Pestalotiopsis microspora HF 12440, an endophytic fungus from the stem of Artocarpus heterophyllus. The structures of all compounds were elucidated using spectroscopic methods and by comparison with the literature. Compound 1 was isolated from the fungi for the first time, compounds 2 and 3 were firstly obtained from this endophytic fungus. Compound 3 showed cytotoxicity (IC50 2.56 µg/mL) against murine leukemia P-388 cells.

Monitoring Metabolite Profiles of Cannabis sativa L. Trichomes during Flowering Period Using 1H NMR-Based Metabolomics and Real-Time PCR.

Cannabis sativa trichomes are glandular structures predominantly responsible for the biosynthesis of cannabinoids, the biologically active compounds unique to this plant. To the best of our knowledge, most metabolomic works on C. sativa that have been reported previously focused their investigations on the flowers and leaves of this plant. In this study, (1)H NMR-based metabolomics and real-time PCR analysis were applied for monitoring the metabolite profiles of C. sativa trichomes, variety Bediol, during the last 4 weeks of the flowering period. Partial least squares discriminant analysis models successfully classified metabolites of the trichomes based on the harvest time. Δ (9)-Tetrahydrocannabinolic acid (1) and cannabidiolic acid (2) constituted the vital differential components of the organic preparations, while asparagine, glutamine, fructose, and glucose proved to be their water-extracted counterparts. According to RT-PCR analysis, gene expression levels of olivetol synthase and olivetolic acid cyclase influenced the accumulation of cannabinoids in the Cannabis trichomes during the monitoring time. Moreover, quantitative (1)H NMR and RT-PCR analysis of the Cannabis trichomes suggested that the gene regulation of cannabinoid biosynthesis in the C. sativa variety Bediol is unique when compared with other C. sativa varieties.

Analysis of cannabinoids in laser-microdissected trichomes of medicinal Cannabis sativa using LCMS and cryogenic NMR.

Trichomes, especially the capitate-stalked glandular hairs, are well known as the main sites of cannabinoid and essential oil production of Cannabis sativa. In this study the distribution and density of various types of Cannabis sativa L. trichomes, have been investigated by scanning electron microscopy (SEM). Furthermore, glandular trichomes were isolated over the flowering period (8 weeks) by laser microdissection (LMD) and the cannabinoid profile analyzed by LCMS. Cannabinoids were detected in extracts of 25-143 collected cells of capitate-sessile and capitate stalked trichomes and separately in the gland (head) and the stem of the latter. Δ(9)-Tetrahydrocannabinolic acid [THCA (1)], cannabidiolic acid [CBDA (2)], and cannabigerolic acid [CBGA (3)] were identified as most-abundant compounds in all analyzed samples while their decarboxylated derivatives, Δ(9)-tetrahydrocannabinol [THC (4)], cannabidiol [CBD (5)], and cannabigerol [CBG (6)], co-detected in all samples, were present at significantly lower levels. Cannabichromene [CBC (8)] along with cannabinol (CBN (9)) were identified as minor compounds only in the samples of intact capitate-stalked trichomes and their heads harvested from 8-week old plants. Cryogenic nuclear magnetic resonance spectroscopy (NMR) was used to confirm the occurrence of major cannabinoids, THCA (1) and CBDA (2), in capitate-stalked and capitate-sessile trichomes. Cryogenic NMR enabled the additional identification of cannabichromenic acid [CBCA (7)] in the dissected trichomes, which was not possible by LCMS as standard was not available. The hereby documented detection of metabolites in the stems of capitate-stalked trichomes indicates a complex biosynthesis and localization over the trichome cells forming the glandular secretion unit.