Sarcorucinine-D Inhibits Cholinesterases and Calcium Channels: Molecular Dynamics Simulation and In Vitro Mechanistic Investigations.

Acetylcholinesterase (AChE) inhibitors and calcium channel blockers are considered effective therapies for Alzheimer's disease. AChE plays an essential role in the nervous system by catalyzing the hydrolysis of the neurotransmitter acetylcholine. In this study, the inhibition of the enzyme AChE by Sarcorucinine-D, a pregnane type steroidal alkaloid, was investigated with experimental enzyme kinetics and molecular dynamics (MD) simulation techniques. Kinetics studies showed that Sarcorucinine-D inhibits two cholinesterases-AChE and butyrylcholinesterase (BChE)-noncompetitively, with Ki values of 103.3 and 4.66 µM, respectively. In silico ligand-protein docking and MD simulation studies conducted on AChE predicted that Sarcorucinine-D interacted via hydrophobic interactions and hydrogen bonds with the residues of the active-site gorge of AChE. Sarcorucinine-D was able to relax contractility concentration-dependently in the intestinal smooth muscles of jejunum obtained from rabbits. Not only was the spontaneous spasmogenicity inhibited, but it also suppressed K+-mediated spasmogenicity, indicating an effect via the inhibition of voltage-dependent Ca2+ channels. Sarcorucinine-D could be considered a potential lead molecule based on its properties as a noncompetitive AChE inhibitor and a Ca2+ channel blocker.

Abietane diterpenes from Abies spectabilis and their anti-pancreatic cancer activity against the MIA PaCa-2 cell line.

An ethanolic extract of the stem of Abies spectabilis exhibited strong cytotoxicity against MIA PaCa-2 human pancreatic cancer cells preferentially under nutrient-deprived conditions. Therefore, phytochemical investigation of this bioactive extract was carried out, and that led the isolation of ten compounds (1-10) including a new abietane-type diterpene (1). The structure of the new compound (1) was elucidated by combined spectroscopic techniques, including HRFABMS, NMR and quantum ECD calculation. All the isolated compounds were evaluated for their efficacy against MIA PaCa-2 human pancreatic cancer cell line by employing an anti-austerity strategy. Among the tested compounds, dehydroabietinol (5) displayed the most potent activity with a PC50 value of 6.6 µM. Dehydroabietinol (5) was also found to retard the MIA PaCa-2 cell migration under normal nutrient-rich conditions displaying its anti-metastatic potential. Investigation on the mechanism suggested that dehydroabietinol (5) is an inhibitor of the key cancer cell survival Akt/mTOR/autophagy signaling pathway.

Phytochemical Analysis and In Vitro Antioxidant and Antibacterial Activity of Different Solvent Extracts of Beilschmiedia roxburghiana Nees Stem Barks.

Plants have long been considered as a basis of medicines for different indigenous cultures around the globe. They continue as a prominent source of important phytoconstituents which exhibit significant biological activities. In this study, we performed the phytochemical screening, estimation of total phenolic and flavonoids, antioxidants, and antimicrobial activities of the stem bark of Beilschmiedia roxburghiana Nees using different solvents. The total phenolic and total flavonoid contents ranged from 106.73 ± 1.62 mg GAE/g and 99.32 ± 0.66 mg QE/g (methanol extract) to 65.59 ± 1.79 mg GAE/g and 29.98 ± 0.90 mg QE/g (n-hexane extract), respectively. The maximum 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity with a half-maximal inhibitory concentration (IC50) of 39.86 ± 3.69 µg/mL was observed for methanol extract followed by aqueous (IC50 = 43.55 ± 6.16 µg/mL), ethyl acetate (IC50 = 44.30 ± 5.88 µg/mL), dichloromethane (IC50 = 71.50 ± 4.70 µg/mL), and the lowest activity was observed for n-hexane extract. The disc diffusion method revealed that the ethyl acetate extract exhibited relatively higher activity against Salmonella typhi (ZOI = 13 mm), and moderate activities against Shigella sonnei, Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus (ZOI = 12 mm). The methanol and aqueous extracts showed nearly parallel and the n-hexane and dichloromethane extracts exhibited mild antibacterial activities. The results indicated that the polarity index of the extracting solvents amplified the biological activities of the extract. The study is helpful to support the validity of the traditional application of the plant as natural medicine.

Potent Insulin Secretagogue from Scoparia dulcis Linn of Nepalese Origin.

Ethno-botanical inspired isolation from plant Scoparia dulcis Linn. (Sweet Broomweed) yielded six compounds, coixol (1), glutinol (2), glutinone (3), friedelin (4), betulinic acid (5), and tetratriacontan-1-ol (6). There structures were identified using mass and 1D- and 2D-NMR spectroscopy techniques. Compounds 1-6 were evaluated for their insulin secretory activity on isolated mice islets and MIN-6 pancreatic ß-cell line, and compounds 1 and 2 were found to be potent and mildly active, respectively. Compound 1 was further evaluated for insulin secretory activity on MIN-6 cells. Compound 1 was subjected to in vitro cytotoxicity assay against MIN-6, 3T3 cell lines, and islet cells, and in vivo acute toxicity test in mice that was found to be non-toxic. The insulin secretory activity of compounds 1 and 2 supported the ethno-botanic uses of S. dulcis as an anti-diabetic agent.

Diterpenes from "Pini Resina" and their preferential cytotoxic activity under nutrient-deprived condition.

In the course of our search for anticancer agents based on a novel anti-austerity strategy, we found that the 70 % EtOH extract of "Pini Resina" showed 100 % preferential cytotoxicity at the concentration of 50 microg/mL. Further bioassay-guided fractionation and purification led to the isolation of 15 compounds including one new compound 7-oxo-13 alpha-hydroxyabiet-8(14)-en-18-oic acid (1). Their structures were elucidated on the basis of spectroscopic analysis. Among the isolated compounds, methyl abieta-8,11,13-trien-18-oate (7) showed the most potent preferential cytotoxicity at 10 microg/mL under nutrient-deprived condition.

Antimalarial activity of cassane- and norcassane-type diterpenes from Caesalpinia crista and their structure-activity relationship.

Malaria is one of the most life-threatening infectious diseases worldwide and claims millions of people's lives each year. The appearance of drug-resistance Plasmodium falciparum has made the treatment of malaria increasingly problematic, and thus, it is a dire need to search the new alternatives of current drugs. In the present study, 44 cassane- and norcassane-type diterpenes isolated from Caesalpinia crista of Myanmar and Indonesia were evaluated for their antimalarial activity against the malaria parasite Plasmodium falciparum FCR-3/A2 clone in vitro. Most of the tested diterpenes displayed antimalarial activity, and norcaesalpinin E (28) showed the most potent activity with an IC50 value of 0.090 microM, more potent than the clinically used drug chloroquine (IC50, 0.29 microM). Based on the observed results, a structure-activity relationship has been established.

Constituents of Caesalpinia crista from Indonesia.

Ten new furanocassane-type diterpenes named, caesalpinins H-P (1-9) and norcaesalpinin F (10), were isolated from the CH(2)Cl(2) extract of the seed kernels of Caesalpinia crista, together with 13 known diterpenes. Their structures were determined based on the spectroscopic analysis. Among the isolated compounds, caesalpinin N (7) represents the first example of furanocassane-type diterpene possessing an aldehyde group at C-14.

Methyl migrated cassane-type furanoditerpenes of Caesalpinia crista from Myanmar.

From the CH2Cl2 extract of seed kernels of Caesalpinia crista from Myanmar, two rare and biogenetically interesting methyl migrated cassane-type furanoditerpenes [caesalpinins MM (1) and MN (2)] and two normal cassane-type furanoditerpenes [caesalpinins MO (3) and MP (4)] have been isolated, together with eight known cassane-type diterpenes, 1-deacetoxy-1-oxocaesalmin C (5), 1-deacetylcaesalmin C (6), caesalmin C (7), bonducellpin C (8), caesaldekarin e (9), 2-acetoxycaesaldekarin e (10), 2-acetoxy-3-deacetoxycaesaldekarin e (11), and norcaesalpinin E (12). Among the known compounds, compounds 5 and 6 were for the first time isolated from a natural source. The structures of these compounds were elucidated by the use of spectroscopic techniques.

New cassane-type diterpenes of Caesalpinia crista from Myanmar.

Seven new cassane-type diterpenes, caesalpinin MF-ML (1-7), and a new norcassane-type diterpene, norcaesalpinin MD (8), have been isolated from the CH2Cl2 extract of seed kernels of Caesalpinia crista from Myanmar, together with sixteen known cassane-type diterpenes, 7-acetoxybonducellpin C, caesaldekarin e, caesalmin C, caesalmin G, 2-acetoxycaesaldekarin e, zeta-caesalpin, caesalpinin D, caesalpinin E, caesalpinin F, caesalpinin H, caesalpinin I, caesalpinin J, caesalpinin K, caesalpinin M, caesalpinin N, and caesalpinin O. The structures of the isolated compounds were elucidated by the use of spectroscopic techniques.