Subagus Wahyuono

Screening of several Indonesian medicinal plants for their inhibitory effect on histamine release from RBL-2H3 cells

Twelve alcoholic extracts and 12 hexane extracts of plant materials selected on the basis of medicinal folklore for asthma treatment in Indonesia were studied for their activity in inhibiting histamine release from RBL-2H3 cells (rat basophilic leukemia cell line), a tumor analog of mast cells. The results of screening indicated that five alcoholic extracts (Plantago major leaves, Eucalyptus globulus leaves and fruit, Cinnamomum massoiae cortex, Vitex trifolia leaves) and two hexane extracts (Eucalyptus globulus leaves, Vitex trifolia leaves) inhibited IgE-dependent histamine release from RBL-2H3 cells. The inhibitory effects were found to be more than 80% for extract concentrations of 0.5 mg/ml. The results indicate that the extracts contain active compounds that inhibit mast-cell degranulation, and provide insight into the development of new drugs for treating asthma and/or allergic disease.

Antiviral and anticancer optimization studies of the DNA-binding marine natural product aaptamine

Aaptamine has potent cytotoxicity that may be explained by its ability to intercalate DNA. Aaptamine was evaluated for its ability to bind to DNA to validate DNA binding as the primary mechanism of cytotoxicity. Based on UV–vis absorbance titration data, the Kobs for aaptamine was 4.0 (±0.2) × 103 which was essentially equivalent to the known DNA intercalator N‐[2‐(diethylamino)ethyl]‐9‐aminoacridine‐4‐carboxamide. Semi‐synthetic core modifications were performed to improve the general structural diversity of known aaptamine analogs and vary its absorption characteristics. Overall, 26 aaptamine derivatives were synthesized which consisted of a simple homologous range of mono and di‐N‐alkylations as well as some 9‐O‐sulfonylation and bis‐O‐isoaaptamine dimer products. Each product was evaluated for activity in a variety of whole cell and viral assays including a unique solid tumor disk diffusion assay. Details of aaptamine’s DNA‐binding activity and its derivatives’ whole cell and viral assay results are discussed.

n-Hexane Insoluble Fraction of Plantago lanceolata Exerts Anti-Inflammatory Activity in Mice by Inhibiting Cyclooxygenase-2 and Reducing Chemokines Levels

Inflammation is involved in the progression of many disorders, such as tumors, arthritis, gastritis, and atherosclerosis. Thus, the development of new agents targeting inflammation is still challenging. Medicinal plants have been used traditionally to treat various diseases including inflammation. A previous study has indicated that dichloromethane extract of P. lanceolata leaves exerts anti-inflammatory activity in an in vitro model. Here, we examined the in vivo anti-inflammatory activities of a n-hexane insoluble fraction of P. lanceolata leaves dichloromethane extract (HIFPL). We first evaluated its potency to reduce paw edema induced by carrageenan, and the expression of the proinflammatory enzyme, cyclooxygenase (COX)-2, in mice. The efficacy of HIFPL to inhibit COX-2 was also evaluated in an in vitro enzymatic assay. We further studied the effect of HIFPL on leukocytes migration in mice induced by thioglycollate. The level of chemokines facilitating the migration of leukocytes was also measured. We found that HIFPL (40, 80, 160 mg/kg) demonstrated anti-inflammatory activities in mice. The HIFPL reduced the volume of paw edema and COX-2 expression. However, HIFPL acts as an unselective COX-2 inhibitor as it inhibited COX-1 with a slightly higher potency. Interestingly, HIFPL strongly inhibited leukocyte migration by reducing the level of chemokines, Interleukine-8 (IL-8) and Monocyte chemoattractant protein-1 (MCP-1).

In Vitro Test and Molecular Docking of Alkaloid Compound in MarineSponge Cinachyrella anomala against T47D Cell Cycle

The compound 1,4,9-triazatricyclo[7,3,1,0]trideca-3,5(13),10-trien-8-ol (SA2014) was isolated from the marine sponge Cinachyrella anomala. In vitro assay for SA2014 compound was found to be able to induce cell-cycle arrest at the sub-G1 and G2/M phases of T47D cancerous cell. A combined dosage between of SA2014 compound and of doxorubicin was able to induce cell-cycle arrest at sub-G1 and G2/M phases. Molecular docking approach showed that SA2014 compound inhibited cdk2 enzyme. The strength of interaction between SA2014 and cdk2 (docking score = -65,43) was more stable than the interaction between doxorubicin and cdk2 (-36,59).

Effects of dihydrocubebin, a lignan isolated from Indonesian plant Piper cubeba, on the histamine release from rat mast cells

The fruits of Piper cubeba L. are used traditionally to treat respiratory disorders in Indonesia. In order to determine the compounds responsible for this activity, the fruits were extracted with nhexane followed by ethanol to give n-hexane and ethanol extracts. Based on tracheospasmolytic assay on these two extracts, the n-hexane extract was more active to inhibit trachea contraction than that of ethanol extract. Upon bioassay guided isolation of the n-hexane extract, a tracheospasmolytic active compound was isolated and identified as dihydrocubebin [(3,4),(3’,4’)-bis-methylenedioxy9,9’dihydroxylignan] (1). Compound 1 was tested further for its ability to inhibit histamine released from mast cells, using rat basophilic leukemia (RBL-2H3) cell line and rat peritoneal mast cells RPMCs) as models; and DNP24-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine released from mast cell. The test result showed that 1 inhibited histamine release from RBL-2H3 cells induced by DNP24-BSA, thapsigargin and ionomycin. In addition, 1 suppressed histamine release from RPMC induced by either thapsigargin or ionomycin. However, 1 did not inhibit histamine release from RPMC induced by either compound 48/80 or combination PMA-sub optimum dose of ionomycin. Therefore, it was concluded that the inhibitory effects of 1 on the histamine released from mast cells may involve mechanisms related to intracellular Ca2+ signaling events or downstream processes of intracellular Ca2+ signaling in mast cells.

MOLECULAR MODELING OF VITEOSIN-A, A TRACHEOSPASMOLYTIC COMPOUND ISOLATED FROM N-HEXANE EXTRACT OF THE LEAVES OF Vitex Trifolia L.

Viteosin-A, a tracheospasmolytic compound, was successfully isolated from n -hexane extract of the leaves of Vitex trifolia L. With the concentration of 0.05 and 0.15 mg/ml viteosin-A inhibited a guinea pig tracheal contraction due to histamine (10-7 – 10-3 M) in vitro by 27.1 and 47.9 %, respectively. Confirmation of C-5 and C-6 configuration is necessary to determine the active reaction site of viteosin-A and its receptor for future development. This research was focused on a molecular modeling of viteosin-A using computational method with HyperChem Pro 4.0 for Windows as software. Based on spectroscopic data and molecular modeling, viteosin-A has S configuration at C-5 and C-6, and therefore was confirmed as (5 S ,10 S )-6 S -acetoxy-8 R -methyl-9-hidroxy-labda-13 Z -en-16,15-olide. Keywords : viteosin-A, molecular modeling

Isolation of aphrodisiac active fraction from sanrego bark (Lunasia amara Blanco)

Sanrego ( Lunasia amara Blanco) bark is traditionally utilized as an aphrodisiac although scientifically has yet been proven. Therefore, this study is aimed to prove aphrodisiac activity, determine the compounds and the dose of the active fraction. Initially sanrego bark (600 g) was extracted in a Soxhlet apparatus with methanol (MeOH) to give MeOH extract (A, 80 g). The MeOH extract was fractionated by Ethylacetate (EtOAc) to give EtOAC soluble (B, relatively non-polar, 15.23 g) and EtOAc insoluble (C, relatively polar, 50.20 g). The aphrodisiac test was performed in male Wistar rats that were divided into 6 groups (5 rats each) [I, treated with Na-CMC 0.5%, 50 mg/kg BW; II, yohimbine, 5 mg/kg BW; III, distilled water , 2 ml/200 g BW; IV, extract A; V, fraction B; VI, fraction C], and the doses given to groups IV-VI were similar (10, 50, 100, and 200 mg/kg BW). The male’s behaviors to female rats ( introduction, climbing and coitus ) were recorded and analysed at p= 0.5. Determination of the active compounds were performed by thin layer chromatography (TLC) using various detection reagents. The result indicated that the highest aphrodisiac effect was demonstrated by fraction B, followed by extract A and fraction C . Fraction B demonstrated introduction (84.2%), climbing (84.9%) and coitus (85.2%). TLC profile suggested that fraction B contain alkaloids and terpenoids as the main components. Keywords: Sanrego bark, Lunasia amara , Aphrodisiac, Active fraction .