Arborine (I), a quinazolone alkaloid was isolated from the leaves of Glycosmis pentaphylla (Rutaceae) and was found to have a growth inhibitory effect on the larvae of Drosophila melanogaster

Ahmad, Farediah

Mohd Ismail, Hazar

Rahmani, Mawardi

Universiti Putra Malaysia Institutional Repository

Pertanika J. Sci. & Techno!. 4(1): 11-15 (1996)ISS :0128-7680© Penerbit Universiti Pertanian MalaysiaArborine, a Larval Growth Inhibitor fromGlycoslnis pentaphyllaFarediah AhDlad,l Hazar B. Mohd ISDlail2 andMawardi RahDlani31Department of ChemistryUniversiti Teknologi MalaysiaSkudai, 80990 Johor Bahru, Malaysia2Centre for Foundation Studies in SciencesUniversiti Malaya59100 Kuala Lumpur, Malaysia3Department of ChemistryUniversiti Pertanian Malaysia43400 Serdang, Selangor, MalaysiaReceived 1 July 1994ABSTRAKArborina (1), suatu alkaloid kuinazolon telah diasingkan daripada daunGlycosmis pentaphylla (Rutaceae) dan didapati memberikan kesan perencatpembesaran ke atas larva Drosophila melanogaster.ABSTRACTArborine (I), a quinazolone alkaloid was isolated from the leaves ofGlycosmis pentaphylla (Rutaceae) and was found to have a growth inhibitoryeffect on the larvae of Drosophila melanogaster.Keywords: GlycOSTnis pentaphylla, Rutaceae, arborine, larvalgrowth inhibitor, Drosophila TnelanogasterINTRODUCTIONIn the course of our search for insect development inhibitors in plants, wescreened the EtOH extracts of dried leaves of Glycosmis pentaphylla Correa bythe Drosophila test (Nakajima and Kawazu 1977). We found that the EtOHextracts of G. pentaphylla inhibited the larval growth of fruit-fly (Drosophilamelanogaster). G. pentaphylla, of the Rutaceae family, is a well-known plant inSouth East Asia. It is used in traditional preparations to treat facialinflammation, itchiness, intestinal problems and coughs (Perry 1980). Inthis paper we report the isolation of the active compound, arborine (1) andits development inhibition activity against the fruit-fly.Farediah Ahmad, Hazar B. Mohd Ismail and Mawardi RahmaniMATERIALS AND METHODS.GeneralUV spectrum was recorded on a Hitachi 200-20 spectrophotometer. IRspectrum was recorded on a Perkin Elmer 727B spectrophotometer. NMRspectra were recorded on a Bruker 300 MHz machine operating at 300MHze H) and 75 MHze3 C). Samples were dissolved in CDC13 containingTMS as an internal standard. TLC were carried out on Merck pre-coatedsilica gel F254 plates while column chromatography was performed usingTLC grade silica gel without binder.Plant MaterialLeaves of G. pentaphylla were collected from Kelantan in early 1990. Avoucher specimen was deposited at the Herbarium, Department of Botany,Universiti Kebangsaan Malaysia, Bangi, Selangor.Extraction and IsolationThe powdered air-dried leaves ofG. pentapfrylla (667.3 g) were defatted withpetroleum ether and then extracted with ethanol (4L X 3) at roomtemperature for 24 h. After removing the solvent, the residue was extractedwith HCl (1 N) which was then neutralised (NaOH) and extracted withCHCI3. The evaporation of the CHC13 layer gave a thick oily residue(3.2 g). Treatment with ethyl acetate - methanol mixture gave a crystallinesolid which was then recrystallised from the same solvent (2 : 8) to give (1).Arborine (1) was isolated as white needles (0.6 g, 0.1 %), m.p l60-16l o C(Chakravarti et al., 1961); UV Amax (EtOH): 245, 260, 277, 284 and 306nm; IR V max (KBr): 3560, 3370,1650,1610,1530 and 1490 cm-\ IH-NMR8 (CDC13): 3.60 (s, 3H, N-CH3 ), 4.26 (s, 2H, -CH2Ph), 7.27(dd, J = 8.6,1.3 Hz, lH, H-8), 7.29(s, 5H, Ph), 7.44(t, J = 7.8 Hz, lH, H-6), 7.68(ddd,J = 8.6, 7.8, 1.3 Hz, lH, H-7), 8.35(dd, J = 7.8, 1.3 Hz, lH, H-5); 13C_NMR ppm: (Table 1); MS m/z (%): 250 (47), 249 (100), 149(10), 133(21),132(14),125 (15),117(13),106(9),105(22),92(15),91 (28) and 90 (15).BioassayThe larval growth inhibitory activity of D. melanogaster was assayed by theDrosophila test (Kawazu et al. 1977). Ten newly-hatched lst-instar larvaewere introduced into the diet (2 g) [ a mixture of dry yeast powder (20 g),honey (40 g), agar (3 g), n-butyl p-hydroxybenzoate (0.3 g) and water(250 g)] containing the test material in a 30 ml sample vial (¢ 25 mm x55 mm) and left for two weeks at room temperature. The development stateof the larvae was observed, and the number of survivors at successive stagesof development recorded every other day and compared with that of acontrol. The assay was carried out in five duplicates and at gradientconcentrations.12 Pertanika J. Sci. & Techno\. Vol. 4 No.1, 1996Arborine, a Larval Growth Inhibitor from Glycosmis pentaphyllaRESULTS AND DISCUSSIONThe alkaloid arborine 1 was first isolated from the leaves of Glycosmis arboreaand its structure was further confirmed as 2-benzyl-1-methylquinazol-4-onebased on UV, IR and IH_ MR data (Chakravarti etal., 1961). Compound1 is optically inactive with a molecular formula C16H14N20, and givesintense peaks at mjz 249 (M+ - 1), 133 and 91 in the MS spectrum. Wewould like here to update the spectral data of 1 since the last reported data(Chakravarti et al. 1961) were incomplete.The 1H-NMR spectrum of 1 showed the presence of two singlets at83.60 and 4.26 due to the N-CH3 and benzylic methylene protons,respectively. A singlet at 8 7.29 was assigned to the five protons of theisolated phenyl nucleus. A doublet of doublet at 8 7.27 which was partlymasked by the phenyl nucleus signal and a triplet centred at 8 7.44 (J = 7.8Hz) were assigned to the protons at C-8 and C-6, respectively. Protonresonances ofH-7 occurred as a doublet ofdoublet of doublet at 8 7.68 (J =8.6,7.8 and 1.3 Hz). Another doublet of doublet at 8 8.35 (J = 7.8 and 1.3Hz) was assigned to the deshielded aromatic proton in the peri-position tothe carbonyl group of a quinazolone system. The 13C-NMR spectrumshowed the presence of 16 carbons with 4 quartenary carbons at 8 169.0,162.1, 141.5 and 120.0. The spectrum also showed the presence of onemethylene carbon which was confirmed by DEPT experiment. Completeassignments for the 13C-NMR chemical shifts of arborine were made bycomparison with the data of similar compounds (2), (3) and (4) for both theheterocylic and the isolated phenyl part of the molecule (Table 1) (Dreyerand Brenner 1980).~1 '0"8 I/~CH 3(1 )(2)Pertanika J. Sci. & Technol. Vol. 4 No.1, 1996 13Farediah Ahmad, Hazar B. Mohd Ismail and Mawardi RahmaniTABLE 113C-NMR chemical shift data for (1), (2), (3) and (4) in ppmCarbon no.245678910N-CH3CH2I'2'3'4'5'6'155.2162.2126.9126.1134.4126. I149.5121.2141.616I.7127.2122.3135. I114.5150.2115.529.4140.4161.4128.9122.8134.91I3.3150.7155.430.6138.6128.9128.4126.4128.4128.9(1)162.2169.0128.5125.9133.7114.4141.5120.034.843.4134.5128.2b129. Ib127.4129. Ib128.2b• From reference 2b Assignment may be interchangedThe growth inhibition activity of arborine is shown in Table 2. The 1st-ins tar larvae were introduced into the diet containing arborine. Arborinecompletely retards the larval growth, resulting in very small andabnormally shaped larvae and pupae. The minimum concentration forthe inhibitory effect on the 1st instar larvae of D. melanogaster was 0.8 mg/2 gdiet (Table 2). However, at lower concentrations, arborine has no effect onthe larval growth. From these results, we conclude that arborine haspotential as a larval growth inhibitor.TABLE 2Larval growth inhibitory activity of arborine (1) against Drosophila melanogasteraDays 2 3 4 6 8 10 12Dose/Diet (mg/2g)4.0 5Ls 5Ls 4Ls 2Ls 0 0 03.0 7Ls 6Ls 5Ls 3Ls 2Ls 0 02.5 8L 7Ls 5Ls 3Ls ILs 0 00.8 8L 5L 5L 4Ps 2Ps 2Ps 00.6 8L 8L 8L 2P6L 3A5P 6A2P 8A0.2 8L 8L 8L 4P4L 4A4L 7AIP 8AControl 10L 10L IOL lOP lOA lOA lOA'L = larvae P = pupae A = adult s = small14 Pertanika J. Sci. & Technol. Vol. 4 No.1, 1996Arborine, a Larval Growth Inhibitor from Glycosmis prntaphyllaApart from quinazolone, other alkaloids such as foruquinoline, acridoneand carbazole are known to be distributed in Glycosmis species (Sarkar andChakraborty 1977; Sarkar Chakraborty 1978). We will further test thealkaloids from this genus to evaluate their effectiveness to control insect pest.AKNOWLEDGEMENTSWe wish to express our thanks to Prof. K. Kawazu of Okayama Universityfor his interest in this work and for obtaining the MS spectrum; Prof SamTeng Wah of Universiti Sains Malaysia for the IH and 13C-NMRmeasurements; Prof. Latif Muhamad and Mr. A. Zainuddin Ibrahim ofUniversiti Kebangsaan Malaysia for identifying and supplying the plantmaterial. Financial assistance from the Malaysian Government under theIRPA .programme is gratefully acknowledged.REFERENCESCHAKRAVARTI, D., R.N. CHAKRAVARTI, L.A. COHEN, D. DASGUPTA, S. DATTAand H.K. MILLER. 1961. Alkaloids of Glycosmis arborea -II Structure of arborine.Tetrahedron 16: 224.DREYER, D.L. and R.C. BRENNER. 1980. Alkaloids of some Mexican Zanthoxyllumspecies. Phytochemistry 19: 935.NAKAJIMA, S. and K. KAWAZU. 1977. Tridec-1-ene-3,5,7,9,ll-pentayne, an ovicidalsubstance from Xanthium canadense. Agric. Biol. Chem. 41(9): 1801-1802.PERRY, L.M. 1980. Medicinal Plants of East and Southeast Asia. p. 366. Massachusetts:MIT Press.SARKAR, M. and D.P. CHAKRABORTY. 1977. Some minor constituents from Glycosmispentaphylla. Phytochemistry 16: 2007.SARKAR, M. and D.P. CHAKRABORTY. 1978. Glycophymoline, a new minor quinazo1inealkaloid from Glycosmis pentaphylla. Phytochemistry. 17: 2145.Pertanika J. Sci. &·Technol. Vol. 4 No. I, 1996 15

Arborine, a Larval Growth Inhibitor from

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