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ANTIFUNGAL EFFECTS OF POMIFERIN ISOLATED FROM MACLURA AURANTIACA FRUIT GROWING IN KAZAKHSTAN
© Orazbekov Y.K.*
National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, Mississippi, USA Kazakh national medical university name of S.D. Asfendiyarova, Almaty, Kazakhstan
South Kazakhstan pharmaceutical academy,
Shymkent, Kazakhstan
Was isolated compound from the Maclura aurantiaca (L.) fruits growing in Kazakhstan. The isolated compound was chemically identified to be Po-miferin. Pomiferin showed a potent antifungal activity against Candida glabrata with IC50 values of 5.7 gg/m. The isolated compound was reported for the first time from Maclura aurantiaca (L.) growing in Kazakhstan.
Keywords: Candida glabrata; prenylated flavonoids; pomiferin, maclura aurantiaca
1. Introduction
Maclura aurantiaca (L.) is known to produce several secondary metabolites belonging to different chemical classes including prenylated flavonoids, which found to have potent pharmacological effects. However, Maclura aurantiaca growing in Kazakhstan has not been examined sufficiently phytochemically or biologically. In this study we report the isolation, characterization and biological activity secondary metabolites isolated from the fruits of Maclura aurantiaca growing in Kazakhstan.
Candida glabrata is one of the most serious causative agents for fungal meningitis and encephalitis, especially as a secondary infection for patients affected by AIDS [1, 2]. Finding a curative agent for Candida glabrata infections is our goal in this research; consequently, biologically guided isolation for the secondary metabolites of Maclura aurantiaca (L.) growing in Kazakhstan has been conducted.
2. Results and discussion
Antifungal bioassay-guided fractionation and isolation for secondary metabolites from Maclura aurantiaca (L.) growing in Kazakhstan lead to the isolation of
MD, PhD докторант.
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Pomiferin. The isolated compound were chemically identified using different spectroscopic techniques to be Pomiferin [3], Pomiferin was isolated in a good quantity (395 mg) and was chemically identified using 1D and 2D NMR methods (see Figures 1-3) and by comparison with the published scientific data [3]. Pomiferin showed a very potent activity against Candida glabrata with IC50 values of 5.7 pg/ml. These compound were reported for first time from Maclura auran-tiaca (L.) growing in Kazakhstan.
220 210 200 190 100 170 160 150 140 130 120 110 100 90 SO 70 60 SO 40 30 20 10 0 -10
flfppm)
Fig. 2. 'H NMR spectrum for Pomiferin
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ФУНДАМЕНТАЛЬНЫЕ И ПРИКЛАДНЫЕ ИССЛЕДОВАНИЯ
4AE-l^-3 8 a ss (СВ as a , C Я
T T 1 V S V w
f~
/ I ТГ 1 1 1
T7"
\ IT T
Ц 1 \
К 7 4 J гот 11 . tj
I =
13 12 11 10 9 8 7 б 5 4 3 2 1 0 -1
fl (ppm)
Fig. 3. 13C NMR spectrum for Pomiferin
Experimental section Plant material
The fresh fruits of Maclura aurantiaca (L.) (4 Kg) were purchased from Shymkent, Kazakhstan.
Extraction and Isolation
Fresh fruits (4 kg) were cut into small pieces and extracted with MeOH (6 L) at 25 °C. Extracts were concentrated under reduced pressure to yield (450 g). The dried extract was suspended in distilled water,(1 L) then partitioned with ethyl acetate (3 X2L), butanol (3 x2L) to afford an ethyl acetate fraction (7 gm), butanol fraction (150 gm) and water fraction (260 gm). The ethyl acetate fraction showed a promising antibacterial activity against MRSA with an IC50 value of 34.12 pg/ml and Methicillin-sensitive Staphylococcus aureus with an C50 value of 43.72 pg/ml. It showed also antifungal activity against C. neoformans with an IC50 value of 65.94 pg/ml. Both of the butanol and water fractions didn’t show any promising antimicrobial activity. The ethyl acetate fraction was subjected to VLC (vacuum liquid chromatography) eluted with chloroform: methanol increasing polarity to yield five groups (1-5). Group 1(7 gm.) was subjected to a silica gel column using eluent (hexane: ethyl acetate (90:10 - 0:100) resulting ten different subfractions (1-10). Subfraction 9 was subjected to sephadex LH-20 column eluted with MeOH, to give compound Pomiferin (395 mg).
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General Experimental Procedures
NMR spectra were recorded on Varian Mercury 400 MHz spectrometer at 400 (1H) and 100 MHz (13C). The HR-ESI-MS spectra were measured using a Bruker Bioapex-FTMS with electrospray ionization (ESI). Column chromatographic separation was performed on silica gel 60 (0.04-0.063 mm) and sephadex LH-20 (0.25-0.1 mm, Merck). TLC was performed on pre-coated TLC plates with silica gel 60 F254 (0.2 mm, Merck).
Antimicrobial assay
Crude extracts and isolated compounds were tested for antimicrobial activity against Candida albicans ATCC 90028, Candida glabrata ATCC90030, Candida krusei ATCC 6258, Aspergillus fumigatus ATCC 204305, Methicillin-resistant Staphylococcus aureus ATCC 33591, Cryptococcus neoformans ATCC 90113, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 35218, Pseudo-monus aeruginosa ATCC 27853, and Mycobacterium intracellulare ATCC 23068, Ciprofloxacin and Amphotericin B were used as positive standards [4].
Conclusions
Here in this research a very potent antifungal secondary metabolite (Po-miferin) against Cryptococcus neoformance with IC50 value less than 0.8 pg/ml, has been isolated with a larg amount (395 mg) for first time from Maclura auran-tiaca (L.) growing in Kazakhstan. This work should be continued with further in vitro studies in purpose to prepare a suitable dosage form for a potent natural antifungal preparation.
Acknowledgment
We are grateful to the Kazakh Government and National Center for Natural Products Research, School of Pharmacy, University of Mississippi, for their financial support.
References:
1. Buchanan K.L., Murphy J.W. What makes Cryptococcus neoformans a pathogen? // Emerging infectious diseases. - 1998. - Vol. 4, № 1. - P. 71.
2. Alvarez M. et al. The outcome of Cryptococcus neoformans intracellular pathogenesis in human monocytes // BMC microbiology. - 2009. - VOl. 9, № 1. - P. 51.
3. Son I.H. et al. Pomiferin, histone deacetylase inhibitor isolated from the fruits of Maclura pomifera // Bioorganic & medicinal chemistry letters. - 2007. -Vol. 17, № 17. - P. 4753-4755.
4. Radwan M.M. et al. Sepicanin A- a new geranyl flavanone from Artocar-pus sepicanus with activity against methicillin-resistant Staphylococcus aureus (MRsA) // Phytochemistry Letters. - 2009. - Vol. 2, № 4. - P. 141-143.
