The genus Amberboa belongs to the family Compositae and comprises six species. One of these is Amberboa ramosa JAFRI, which is an annual herbaceous plant found in India and Pakistan. The plant has tonic, aperient, febrifuge, deobstruent, cytotoxic, and antibacterial activities.1) Previously triterpenoids, flavanoids, steroids, and steroidal glycosides have been reported from this species.1,2) Here we report the isolation and structure elucidation of long-chain esters 1 and 2, along with two known compounds tricontane (3) 3) and apigenin (4).
4)Tyrosinase (EC 1.14.18.1) is a multifunctional copper-containing enzyme widely distributed in plants and animals. It catalyzes the oxidation of monophenols, o-diphenols, and o-quinones. Tyrosinase is known to be a key enzyme for melanin biosynthesis in plants and animals. Tyrosinase inhibitors therefore can be clinically useful for the treatment of some dermatologic disorders associated with melanin hyperpigmentation. They also find use in cosmetics for whitening and depigmentation after sunburn. It has recently been shown that other factors such as metal ions and the TRP-1 and TRP-2 enzymes also contribute to the production of melanin. However, tyrosinase plays a critical regulatory role in melanin biosynthesis. Therefore many tyrosinase inhibitors that suppress melanogenesis have been actively studied with the aim of developing preparations for the treatment of hyperpigmentation. 5) Compounds 1 and 2 showed strong to moderate inhibitory activity against tyrosinase.
Results and DiscussionThe chloroform-soluble fraction of the whole plant of A. ramosa was subjected to column chromatography over flash silica eluted with different mobile phases. Compounds 1-4 were finally obtained and their structures established by IR, mass, and NMR spectroscopy including 2D NMR techniques. Compound 1 was isolated as a colorless powder. The molecular formula C 34 H 66 O 3 was determined based on the HR-EI-MS, FD, and 13 C-NMR spectra. HR-EI-MS showed a molecular ion peak at m/z 522.5017 (Calcd for C 34 ϩ indicated a double bond at C-7. The losses of 31 and 29 mass units from the [M] ϩ peak were due to the elimination of methoxy and ethyl groups, respectively. The position of the double bond was also confirmed by oxidative cleavage with osmium tetraoxide which gave a mixture of products, among which hexacosanal could be isolated and identified. The 1 H-NMR spectrum displayed a triplet at d 0.85 (Jϭ7.0 Hz) and a broad singlet at d 1.23 typical of a straight-chain hydrocarbon. It also showed signals at d 5.40 (1H, dt, Jϭ16.3, 6.1 Hz), 5.37 (1H, dt, Jϭ16.3, 6.1 Hz), 3.94 (m), and 3.85 (br s) for olefinic protons, aH (H-2), and methoxy protons, respectively. The 13 C-NMR spectrum (BB and DEPT) showed signals at d 175.6, 130.7, and 129.6 which could be assigned to the carbonyl and olefinic carbons, respectively. One oxygenated methine and a methoxy carbon resonated at d 72.2 and 51.4, respectively. The signals observed between 29.0-29.6 indicated the presence of a long chain. The stereochemis...