CC BY
32
АНТИМИКРОБНЫЙ ЭФФЕКТ ОСАЙИНА ВЫДЕЛЕННЫЙ ИЗ ПЛОДОВ MACLURA AURANTIACA, ПРОИЗРАСТАЮЩЕЙ В КАЗАХСТАНЕ
© Оразбеков Е.К.*
Национальный центр по исследованию натуральных продуктов
школы фармации, Университет Мисиссиппи, США, Миссиссиппи Казахский национальный медицинский университет имени С.Д. Асфендиарова, Республика Казахстан, г. Алматы Южно-Казахстанская государственная фармацевтическая академия, Республика Казахстан, г. Шымкент
Было выделено соединение из плодов Maclura aurantiaca (L.) произрастающий в Казахстане. Выделенное соединение был химически идентифицирован как Осайин. Осайин показал очень высокую активность против метициллин-устойчивого золотистого стафилококка (MRSA) с IC50 значением меньше чем в 0,12 мкг / мл. Это соединение было представлено впервые от плодов Maclura aurantiaca (L.) произрастающей в Казахстане.
Ключевые слова: MRSA, prenylated flavonoids, осайин, maclura aurantiaca.
1. Introduction
Several types of compounds have been isolated from Maclura aurantiaca (L.) Gaud. [1]. Prenylated flavonoids are one of most interesting compounds found in of compounds have been isolated from Maclura aurantiaca sp. because of their potent pharmacological properties. However, Maclura aurantiaca growing in Kazakhstan has not been sufficiently examined for its phytochemical or biological properties. In this study we report the isolation, characterization and biological evaluation for Osajin isolated from the fruits of Maclura aurantiaca.
MRSA is a bacterium responsible for several difficult-to-treat infections in humans due to its resistance to many antibiotics and is more dangerous if it is accompanied with an infection of other types of bacteria such as acanthamoeba [2, 3].
On 8 June, at 2011 an E.coli outbreak in Europe cost the EU $2,840,000,000 in human losses (such as sick leave) and about $210,000,000 in material losses [5].
Exploring a curative agent for MRSA and E. coli is one of our projects; consequently, biologically guided isolation for the secondary metabolites of Maclura aurantiaca ” (L.) fruits growing in Kazakhstan has been conducted.
2. Results and discussion
Antimicrobial bioloassay-guided fractionation and isolation of secondary metabolites from Maclura aurantiaca (L.) fruits growing in Kazakhstan lead to the isolation of 6 compounds. The isolated compound was chemically identified using different spectroscopic techniques to be Osajin [6].
MD, PhD докторант.
Инновации в химии, фармацевтике, медицине
93
Osajin was isolated with in a good quantity (140 mg) and chemically identified using 1D and 2D NMR experiments (see Table 1 and Figures 2-5) and by comparing with the published data in the scientific literature [6]. The antimicrobial activities for the compound have been conducted as showen in Table 1. Osa-jin showed a very potent activity against MRSA, Methicillin-sensitive Staphylococcus aureus with IC50 less than 0.12 ug/ml.
Fig. 1. Key HMBC correlations for Osajin
Table 1
Antimicrobial activities for Osajin
Osajin IC50 ng/ml
MRSA <0.12
P. aeruginosa 9.98
M. intracellulare 2.34
C. neofomans >20
94 УПРАВЛЕНИЕ ИННОВАЦИЯМИ: ТЕОРИЯ, МЕТОДОЛОГИЯ, ПРАКТИКА
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 ethyl acetate fraction (7 gm), butanol fraction (150 gm) and water fraction (260 gm). The ethyl acetate fraction showed antibacterial activity against MRSA with an IC50 value of 34.12 pg/ml and Methi-cillin-sensitive Staphylococcus aureus with IC50 value of 43.72 pg/ml. It showed also antifungal activity against C. neofomans with an IC50 value of 65.94 pg/ml. Both 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) in a matter of 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 5 was subjected to Sephadex LH-20 column eluted with MeOH to give compound 1 (140 mg).
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
Инновации в химии, фармацевтике, медицине
95
Bruker Bioapex-FTMS with electrospray ionization (ESI). Column chromatographic separation was performed on silica gel 60 (0.04-0.063 mm) and sepha-dex LH-20 (0.25-0.1 mm, Merck). TLC was performed on precoated 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, Asperigillus fumigatus ATCC 204305, Methicillin-resistant Staphylococcus aureus ATCC 33591, Cryptococcus neoformans ATCC 90113, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Mycobacterium intracellulare ATCC 23068, Ciprofloxacin and Amphotericin B were used as positive standards.
Conclusions
Here in this research a potent anti MRSA and anti E. coli secondary metabolite (Osajin) has been isolated with a large amount (140 mg) for first time from Maclura aurantiaca (L.) fruits growing in Kazakhstan. This work can considered as a good preliminary study for further in vitro studies and preparing a suitable dosage form for a natural antibacterial preparation.
Acknowledgment
We are grateful to the Kazakh Government and National Center for Natural Products Research (NCNPR), School of Pharmacy, University of Mississippi, for their financial support.
References:
1. Grandtner M. M. Elsevier's Dictionary of Trees: Volume 1: North America. -Elsevier, 2005. - Vol. 1.
2. Lone R. et al. Unusual case of methicillin resistant Staphylococcus aureus and Acanthamoeba keratitis in a non-contact lens wearer from Kashmir, India // BMJ case reports. - 2009. - Vol. 2009. - P. 642.
3. Huws S.A. et al. Amoebae promote persistence of epidemic strains of MRSA // Environmental microbiology. - 2006. - Vol. 8, №. 6. - P. 1130-1133.
4. Junillon T. et al. Simplified detection of food-borne pathogens: an in situ high affinity capture and staining concept // Journal of microbiological methods. -2012. - Vol. 91, № 3. - P. 501-505.
5. Karch H. et al. The enemy within us: lessons from the 2011 European Escherichia coli O104: H4 outbreak // EMBO molecular medicine. - 2012. - Vol. 4, № 9. - P. 841-848.
6. 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.
