PHYTOCHEMICAL AND BIOLOGICAL INVESTIGATION OF CLEOME BRACHYCARPA VAHL. GROWING IN EGYPTHTML Full Text
PHYTOCHEMICAL AND BIOLOGICAL INVESTIGATION OF CLEOME BRACHYCARPA VAHL. GROWING IN EGYPT
M. S. Afifi
Department of Pharmacognosy, Faculty of Pharmacy, MISR International University, Cairo Ismailia Road, Cairo, Egypt.
ABSTRACT: Flavonoid and sesquiterpene constituents of the aerial part of Cleome brachycarpa were chemically investigated and screened for their potential cytotoxicity. A flavonol glycoside; 3, 5, 4` trihydroxy-3`-methoxy flavone - 7 – O - a - L -rhamnopyranose (1```® 2``) - O – β - D-glucopyranoside (1) and a megastigmane glucoside; (+)-(6S, 9R) roseoside (2) were isolated for the first time from the n-butanol fraction of the 90 % ethanol extract of Cleome brachycarpa. Four methylated flavonoids were isolated from the chloroform fraction and identified as 5,4`- dihydroxy-3,6,7,3' tetramethoxyflavone (3), 5,5`- dihydroxy-3,6,7,3',4', pentamethoxy-flavone (4), 5-hydroxyl -3,6,7,3',4',5'-hexamethoxyflavone (5) and 5, 7,3',4' tetrahydroxyflavone (6) In addition, two sesquiterpene oxides, namely buchairol (7) and teucladiol (8) were isolated from the n-hexane fraction for the first time. Structure elucidation was achieved using spectroscopic techniques, including IR, UV, ESI-MS, EI-MS and 1D, and 2D-NMR. Each of the three fractions and isolated compounds were screened for their cytotoxicity and lethality using the brine shrimp (Artemia salina) assay. The results showed high lethality for all the examined samples, which might be very useful as antiproliferative and antitumor.
Cleome brachycarpa, α-flavonol glycoside, Methylated flavonoid, Buchairol, Teucladiol, Brine shrimp
INTRODUCTION: Different localities in Egypt are characterized by the presence of a considerable number of medicinal plants that are highly used in folk treatments. Cleome brachycarpa Vahl belongs to family Cleomaceae or Capparaceae 1. Cleomaceae is a small family of flowering plants in the order Brassicales, comprising more than 300 species belonging to 9 genera of which Cleome is the largest genus with about 180 - 200 species of medicinal, ethnobotanical and ecological importance.
Plants of this family are herbs, shrubs or trees, sometimes-woody climbers. Nine species of genus Cleome are distributed in Egypt 1, 2. Species of Cleome were reported to exhibit several bioactivities and uses, such as antimicrobial 3 hepatoprotective immunomodulatory 4, anticancer 5, antiviral 6 and antioxidant 7. Different chemical classes such as flavonoids 8, 9, 19 coumarino-lignans 11 steroids 12, dammarane-type triterpenes 13,14, trinortriterpenoid dilactone 15, 16, sesquiterpenes 17, 18, bicyclic diterpene 19, and betaines were isolated from different species of genus Cleome.
Leaves of Cleome brachycarpa Vahl. are used for the treatment of rheumatism and as anti-inflammatory 8, 20, antidermatosis (scabies and leucoderma) 20, carminative 20, 21 and anti-emetic 22. Chemically, flavonoids 8, 9, trinortriterpenoid dilactone, deacetoxybrachycarpone, cabralealactone, ursolic acid were reported in the plant 23. The present study aimed to evaluate the main chemical constituents and screening the cytotoxicity of the aerial part of Cleome brachycarpa Vahl. growing in Egypt.
MATERIALS AND METHODS:
Plant Material: The aerial part of Cleome brachycarpa Vahl was collected from Wadi El Gemal National park, Red Sea coast, Egypt, in March 2010. The plant was kindly identified by Dr. M. Gebali (Plant Taxonomy and Egyptian Flora Department, National Research Center, Giza, Egypt). A voucher specimen has been deposited in the herbarium of the Pharmacognosy Department, Faculty of Pharmacy, Misr International University, Cairo, Egypt.
General Experimental Procedure: UV spectra were determined using a Hitachi 340 spectro-photometer; IR spectra were carried out on a Nicolet 205 FT IR spectrometer connected to a Hewlett-Packard Color Pro. Plotter. The 1H- and 13C-NMR measurements were obtained with a Bruker NMR spectrometer operating at 500 MHz (for 1H) and 100 MHz (for 13C) in DMSO-d6 or CDCl3 solution, and chemical shifts were expressed in (ppm) concerning TMS, and coupling constants (J) in Hertz. 1H-13C HMBC NMR experiments were carried out using a Jeol AMX-500 high field spectrometer equipped with software Master nova version 5.1.1-3092 program for NMR. ESI-MS (positive ion acquisition mode) was carried out on a TSQ700 triple quadruple instrument (Finnigan, Santos, CA, USA). TLC was performed on precoated TLC plates with silica gel 60 F254 (layer thickness 0.2 mm, E. Merck, Darmstadt, Germany). Column chromatography was carried out using Silica gel 60 (Merck, 40-63 and 63-200 µ) and Sephadex LH-20 (Sigma, 25-100 µ). Developed chromatograms were visualized by spraying with 1% vanillin/H2SO4 24 or anisaldehyde/ H2SO4 reagent 24 followed by heating at 100 ºC for 5 min.
Extraction and Isolation:
Extraction and Fractionation: The air-dried powdered material (1 Kg) was exhaustively extracted with 90% ethanol (3 × 5 L). The combined ethanol extract was concentrated under vacuum at 40 °C to give a brown residue (175 g). The obtained residue was suspended in distilled H2O (700 ml) and defatted with petroleum ether, then partitioned successively with n-hexane, chloroform, and n-Butanol. Each fraction was concentrated under vacuum at a temperature not exceeding 40 °C to afford (12, 20 and 25 g), respectively.
Isolation of Compounds from n-Butanol Fraction: The n-Butanol fraction (18 g) was purified by chromatography on Sephadex LH-20 (100 × 5 cm, 400 g) using solvents CH2Cl2−MeOH (4:1), CH2Cl2−MeOH (3:2), CH2Cl2−MeOH (1:4) and MeOH to yield four fractions [A (2 g), B (1.8 g), C (3.5 g) and D (4 g)]. Fraction B was further fractionated over a silica gel column (60 g) and eluted with EtOAc and EtOAc/MeOH mixtures in a gradient elution system. Fractions eluted with 15% MeOH in EtOAc gave compound-1 as a light yellow amorphous powder (200 mg) after purification by sephadex LH-20 column with CH2Cl2−MeOH (65:35) as eluent. Fraction D was subjected to column chromatography over silica gel (80 g) and eluted with CHCl3 followed by increasing concentrations of MeOH in CHCl3 (up to 30%) to give three subfractions. The first subfraction (2 g) was further purified by repeated chromatography on Sephadex LH20 column (50 g), using H2O- MeOH mixture to give Compound-2 as an amorphous powder (25 mg).
Isolation of Compounds from the Chloroform Fraction: Chloroform fraction (20 g) was further fractionated by CC (Sephadex LH-20, 400 g) using CH2Cl2−MeOH (4:1), and increasing the polarity till 100% MeOH as eluents afforded, three subfractions [E (7.5 g), F (3.2 g), and G (3.7 g)]. Subfraction E (7.5 g,) was fractionated on a silica gel column (170 g) using 100% CHCl3 as a solvent system and increasing the polarity with MeOH.
Similar eluates were pooled together, and eluates with 8% MeOH-CHCl3 (4 g) were subjected to column chromatography on silica gel (100 g). The column was eluted with n-hexane with increasing the polarity with EtOAc. Repeated chromatography on LiChroprep® RP-18 column (50 g) and elution with 20% H2O-MeOH mixture showed three major spots. Chromatography on sephadex LH20 using 20% H2O-MeOH as a developing solvent gave compound 3 (20 mg), compound 4 (25 mg) and compound 5 (40 mg). Fraction 3 (3.0 g) obtained from chloroform fraction was chromatographed over SephadexLH-20 CC using CH2Cl2−MeOH (7:3) as eluent to give 25 mg of light yellow amorphous powder of compound 6.
Isolation of Compounds from the n-hexane Fraction: The n-hexane fraction (6.0 g) was chromatographed over silica gel column (100 g), eluted with n-hexane and n-hexane/ EtOAc mixtures in a gradient elution. Fraction H eluted with n-hexane: EtOAc (1:3) showed a major violet spot with vanillin- sulphuric acid spray reagent. It was further purified on a silica gel CC eluted with n-hexane with increasing the polarity with EtOAc to produce subfraction I (100 mg) and subfraction J (75 mg). Each subfraction was further purified on sephadex LH-20 CC with CH2Cl2: MeOH (7:3) as eluent to give compound 7 from subfraction I (30 mg) and compound 8 from subfraction J (23 mg).
Compound-1: mp 224-226 °C; HRf 38 (solvent system: 35% MeOH in EtOAc). UV dmax (MeOH): 354, 268 and 254; (NaOMe) 398, 266 and 249 nm; (AlCl3) 398, 362 and 269 nm; (AlCl3/HCl) 401, 359 and 269 nm; (NaOAc) 412 and 262 nm; ESI-MS (m/z 625 [M + 1]+ and 647 [M + Na]+ calc. for C28H32O16); 1H-NMR (500 MHz, DMSO-d6): dH 6.46 (1H, d, J = 1.8 Hz, H-6), 6.85 (1H, d, J = 1.8 Hz), 8.05 (1H, d, J = 2.0 Hz, H-2`), 6.90 (1H, d, J = 8.5 Hz, H-5`), 7.75 (1H, dd, J = 8.5, 2.0 Hz, H-6`), 3.85 (3H, s, OCH3-3`), 5.52 (1H, d, J = 7.6 Hz, C-1``), 4.32 (1H, dd, J = 7.6, 9.0 Hz, H-2``), 3.20 (1H, t, J = 9.0 Hz, H-3``), 3.14 (1H, t, J = 10.2 Hz, H-4``), 3.18 (1H, m, H-5``), 3.48 (1H, dd, J = 11.5, 6.5 Hz, H-6``a) 3.67 (1H, dd, J = 11.5, 3.2 Hz, H-6``b), 5.58 (1H, brs, C-1```), 3.40 (1H, m, H-2```), 3.70 (1H, m, H-3```), 3.35 (1H, m, H-4```), 3.94 (1H, m, H-5```), 0.72 (3H, d, J = 6.1 Hz, H-6```); 13C-NMR (100 MHz, DMSO-d6): dC 161.50 (C-5), 99.85 (C-6), 163.42 ( C-7), 94.77 (C-8), 155.80 (C- 9), 105.77 (C-10), 122.55 (C-1`), 115.40 (C-2`), 148.15 (C-3`), 153.15 (C-4`), 116.65 (C-5`), 123.53 (C-6`), 56.20 (OCH3-3`), 101.20, (C-1``), 77.68 (C-2``), 74.35 (C-3``), 71.80 (C-4``), 78.29 (C-5``), 62.50 (C-6``), 98.70 (C-1```), 70.76 (C-2```), 70.15 (C-3```), 73.40 (C-4```), 70.24 (C-5```), 17.75 (C-6```).
Compound-2: Rf = 0.38, solvent system: 50% H2O−MeOH; [α]D + 78.1 (c 0.025, MeOH); IR (film) dmax 3398, 2968, 2929, 1652, 1436, 1373, 1072 and 1033 cm-1; UV dmax (MeOH): 238 nm; ESI-MS (m/z 409 [M + Na]+ calc. for C19H30O8); 1H NMR (500 MHz, CD3OD): dH 2.19 (1H, d, J = 16.8 Hz, H-2a), 2.68 (1H, d, J = 16.8 Hz, H-2b), 5.82 (1H, s, H-4 ), 5.81 (1H, d, J = 15.6 Hz, H-7), 5.78 (1H, d, J = 15.6 Hz, H-8), 4.41 (1H, dq, J = 6.6 and 6.0 Hz, H-9), 1.20 (3H, d, J = 6.6 Hz, CH3-10), 0.92 (3H, s, CH3-11 ), 1.02 (3H, s, CH3-12), 1.72 (3H, d, J = 1.2 Hz, CH3-13), 4.48 (1H, d, J = 7.8 Hz, H-1`), 3.32 (1H, dd, J = 7.8, 9.3 Hz, H-2`), 3.26 (1H, t, J = 9.3 Hz, H-3`), 3.44 (1H, t, J = 10.5 Hz, H-4`), 3.29 (1H, m, H-5`), 3.65 (1H, dd, J = 12.3, 6.2 Hz, H-6`a) 3.80 (1H, dd, J = 12.3, 2.5 Hz, H-6`b); 13C NMR (100 MHz, CD3OD): dC 41.55 (C-1), 52.0 (C-2), 206.15 (C-3), 127.54 (C-4), 166.40 (C-5), 82.13 (C-6), 132.10 (C-7), 134.60 (C-8), 77.65 (C-9), 20.56 (C-10), 22.40 (C-11), 24.57 (C-12), 21.15 (C-13), 101.25 (C-1`), 74.70 (C-2`), 77.57 (C-3`), 71.23 (C-4`), 78.50 (C-5`), 62.84 (C-6`).
Brine Shrimp Lethality Bioassay: Brine shrimp (Artemia salina) lethality bioassay 25, 26 was carried out to investigate the cytotoxicity of the three fractions (n-hexane, CHCl3 and n-butanol) as well as the eight isolated compounds from Cleome brachycarpa. In each experiment, 10 mg of each of the n-Hexane, chloroform and n-Butanol fractions and 5 mg of isolated compounds [1-8], were dissolved in DMSO and solutions of varying concentrations (10-500 µg/ml) were obtained by serial dilution technique using DMSO, 0.5 ml of each sample was added to 5 ml of brine solution and maintained at room temperature for 24 h under the light and surviving larvae were counted.
Experiments were conducted along with control (solvent-treated). Vincristine sulfate was used as a positive control.
Determination of Lethality Concentration: The percentage of lethality was calculated by comparing the mean survival larvae of fractions and pure isolated compounds treated tubes and control. An approximate linear correlation was observed when the logarithm of concentration versus the percentage of mortality was plotted, and the values of LC50 were calculated using Microsoft Excel® 2007.
Statistical Analysis: The lethal concentration of tested samples resulting in 50% mortality of the brine shrimp LC50 were determined from the 24 h counts, and the dose-response data were transformed into a straight line by means of a trend line fit linear regression analysis (Microsoft Office Excel 2007), and finally the LC50 was derived from the best-fit line obtained.
RESULTS AND DISCUSSION: The 90% ethanol extract of the air-dried aerial part of C. brachycarpa was successively partitioned between n-hexane, chloroform and n-Butanol to give the corresponding soluble fractions. A combination of normal phase Si gel, reversed phase RP-18 Si gel and Sephadex LH 20 column chromatography of each fraction led to the isolation of compounds [1 and 2] from the n-Butanol fraction, [3-6] from chloroform fraction and [7 and 8] from the n-hexane fraction.
Compound  was obtained as an amorphous yellow powder. It gave an intense yellow colour with ammonia vapor and yellowish brown color with vanillin/ H2SO4 spraying reagents. The positive-ion ESI-MS showed a pseudo-molecular ion peak at m/z 625 (M + H)+ and 647 (M + Na)+ which in conjunction with the 13C-NMR spectral data, indicates that its [M]+ was 624, suggesting the molecular formula C28H32O16. The UV absorption maxima recorded in MeOH showed two absorptions at 268 and 354 nm, characteristic for a flavonol skeleton 27 Fig. 1.
The 1H- and 13C-NMR spectra of  showed two meta-coupled protons (AB-spin system) at dH 6.46 and 6.85 (each, 1H, d, J = 1.8 Hz, dC 99.85 (C-6), 94.77 (C-8) of ring A; characterized the 6- and 8-protons of a flavonoid with 5, 7 dihydroxy A-ring. It also showed three aromatic protons at dH 8.05 (1H, d, J = 2.0 Hz, H-2`, dC 115.40), dH 6.90 (1H, d, J = 8.5 Hz, H-5`,dC 116.65) and 7.75 (1H, dd, J = 8.5, 2.0 Hz, H-6`, dC 123.53) and represented ABX spin-pattern able assigned to a disubstituted B-ring of 3`,4'-oxygenated 28 confirmed an aglycone with A-ring functionality at C-5 (d 161.50) and C-7 (d 163.42), and B-ring at C-3`(d 148.15) and C-4` (d 153.15).
The UV bathochromic shift with NaOMe (dmax 398nm), AlCl3/HCl dmax 401, 359, and 269 nm) indicated 3, 5, a 4`free hydroxyl group. The position of the methoxyl group at C3` was indicated from HMBC of its protons at dH 3.85 with C3` at dC 147.4. In addition, the 1H- and 13C-NMR spectra of  showed a methoxyl signal at dH 3.85 (3H, s, dH 56.20, OCH3-3`) and two sugars as concluded from the two anomeric protons at dH 5.52 (1H, d, J = 7.6 Hz, dC 101.20, C-1``) and 5.58 (1H, brs, dC 98.70, C-1```). In 13C-NMR, the methyl carbon signals at dC 17.75, C-6```, and proton signals at dH 0.72 (3H, d, J = 6.1 Hz, H-6```, indicated that  contained a methyl pentose sugar. The chemical shifts of the sugar carbon were in agreement with those reported for O-a-L-rhamnopyranosyl (1```® 2``)-O-β-D-glucopyranoside 28, 29. The sugar units are attached to C-7 position based on the lack of bathochromic shift with NaOAc and the characteristic correlations observed between the glucosyl anomeric proton (dH 5.56) and methine carbons at (C-6 and C-8) and quaternary carbon at (dC 155.80, C- 9). Consequently, the structure of  was established as 3, 5, 4` trihydroxy-3`-methoxy flavone -7-O-a-L-rhamnopyranosyl (1```® 2``)-O-β-D-glucopyranoside Fig. 1, and was in strong agreement with that previously isolated from Cleome droserifolia 8.
Compound  was isolated as an amorphous powder. The molecular formula was determined as C19H30O8 based on the quasi-molecular ion peaks observed at 409 (M + Na) + by ESI-MS. The UV spectrum of  (lmax 238 nm) and the IR absorption (nmax 1652 cm-1) indicated the presence of an a,b-unsaturated ketone Fig. 1.
The IR spectrum also exhibited strong absorptions at 3398 cm-1, indicating the existence of hydroxyl functionalities. The 1H- and 13C-NMR spectra with the aid of 1H-13C-HMBC showed signals for two olefinic protons at dH 5.86 (1H, d, J = 15.2, Hz, H-7, dC 132.10 ) and 5.79 (1H, d, J = 15.2 Hz, H-8, dC 134.60), which indicated the presence of a trans double bond; an oxymethine proton at dH 4.41 (1H, dq, J = 6.6 and 6.0 Hz, H- H-9, dC 77.65) and a secondary methyl group at dH 1.20 (3H, d, J =6.6 Hz, Me-10, dC 20.56), as an AMXY3-type spin system.
Additional signals were found to be due to three tertiary methyl groups at dH 0.92 (3H, s, Me-11, dC 22.40), 1.02 (3H, s, Me-12, dC 24.57) and one being vinylic at dH 1.72 (3H, d, J = 1.2 Hz, Me-13, dC 21.15), a vinyl proton at dH 5.82 (1H, s, H-4, dC 127.54) and one methylene proton signals at dH 2.19 and 2.68 (2H, each d, J = 16.8 Hz, H-2, dC 50.75) were also observed. 1H-13C-HMBC showed correlations between CH3-13 and C-4, C-5 and C-6 which confirmed the location of the conjugated ketone function at C-3, dC 206.15, and the correlations between CH3-11 and CH3-12 and C-1 (dC 41.55), C-2 (dC 52.0) and C-6 (dC 82.13) also clarified the six-membered ring moiety of . Signal multiplicities, chemical shifts and coupling constants in the 1H- and 13C-NMR spectra of  revealed the resonance of an anomeric proton and carbon signals at (dH 4.48, d, J = 7.8 Hz, H-1`, dC 101.25) which were inconsistent with the presence of an β-D- glucopyranosyl unit. A loss of 162 mass units from the molecular-ion in the ESI-MS at m/z 225 (M + H- glucose) + and m/z 207 (M + H- glucose - H2O) + suggested the presence of a glucose moiety in  Fig. 1. The linkage position of the sugar moiety was unambiguously determined to be at C-9 by the long-range correlation from (dH 4.48, H-1') of the sugar moiety to dC 77.65 (C-9) of the aglycone unit in the 1H-13C-HMBC spectrum of .
FIG. 1: STRUCTURE OF ISOLATED COMPOUNDS FROM CLEOME BRACHYCARPA
These spectroscopic data suggested that  is a megastigmane glucoside that was very similar to roseoside (+) - (6S, 9R) – 9 – O - b-D-glucopyranosyloxy-6- hydroxy-3-oxo-aionol)). Compared to the 13C-NMR spectral data of (6S, 9R)-roseoside 27, the chemical shift values assigned to C-6, C-7, C-8, and C-9 in  were similar to those reported for the aglycone moiety of (+)-(6S,9R) of roseoside 30. An upfield shift of C-9 (ca. 74 ppm) is indicative for the (9S)-configuration whereas compounds with (9R)-configuration exhibit a lower field signal (ca. 77 ppm). This empirical rule could be verified thorough comparison with literature data for diastereomeric 3-oxo-a-ionol glucosides 31. The chemical structure of  was proposed as: (+)-(6S,9R)-9-O-b-D-glucopyranosyloxy-6-hydroxy-3-oxo-a-ionol Fig. 1 and is identical to (6S,9R)-roseoside) by literature comparison 30-32 .
All the assignments of the isolated compounds [2-8] Fig. 1, were supported by 1H-C13 NMR, ESI-MS, HMQC and HMBC experiments as well as literature comparison which allowed the identification of compound  and  as 5,4`- dihydroxy-3,6,7,3' tetramethoxy flavone and 5-hydroxyl-3, 6, 7, 3' ,4', 5' - hexamethoxyflavone, respectively previously reported from C. brachycarpa 9. Whereas, Compound 6 was identified as 5, 7, 3', 4' tetrahydroxyflavone (luteolin) previously isolated from Cleome species 8. Compounds  and  were identified as 5, 3`- dihydroxy-3, 6, 7, 5', 4', pentamethoxy-flavone and buchairol, respectively previously isolated from Cleome droserifolia 8, 18, 33. Compounds  was identified as 6-hydroxynardol (teucladiol) which was isolated before from Teucrium leucocladum 34. Compounds (1, 2, 4, 7 and 8) have been isolated for the first time from Cleome brachycarpa. Although Sharaf et al., 8 reported the absence of compound  from C. brachycarpa collected from EI-Taaif, Saudi Arabia in August but the current study reported its presence for the first time in the sample collected from Wadi El Gemal National park, Red Sea coast, Egypt, in March which may be attributed to seasonal, environmental or soil variations.
The brine shrimp lethality assay represents a rapid, inexpensive and simple bioassay for testing plant extracts bioactivity, which in most cases correlates reasonably well with cytotoxic and anti-tumor properties 25, 26. In the present study, the brine shrimp lethalities of the n-Hexane, chloroform, n-Butanol fractions of the 90% ethanol extract of Cleome brachycarpa and isolated compounds [1-8], were determined 25, 26. The LC50 values of the brine shrimp lethality testing after 24 h of exposure of test samples and that of the positive control, vincristine sulfate are given in Table 1.
In comparison with vincristine sulfate, the cytotoxicity assay of the chloroform and n-hexane fractions of Cleome brachycarpa showed significant brine shrimp lethalities with LC50 values 15.10 and 26.50 μg respectively. Also, the n-Butanol fraction exhibited moderate brine shrimp lethality and the LC50 (52.10 μg) value was found to be lower than 100 μg 25, 26. The isolated compounds; (5, 4, 7 and 8) exhibited significant brine shrimp lethalities with LC50 values (LC50<30 μg/ml) 10.10, 12.75, 18.20 and 21.80 μg respectively, while compounds; (3, 1, 2 and 6) showed moderate brine shrimp lethalities with LC50 values 37.00, 40.31, 45.25 and 79.50 μg respectively.
The degree of lethality was found to be directly proportional to the concentration of the tested samples. The LC50 values of the tested samples were obtained by plotting the percentage of the Shrimp nauplii (larva) killed versus the logarithm of concentrations of the extracts or isolated compounds and the best-fit line was obtained from the data using regression analysis.
The presence of significant lethality of C. brachycarpa to brine shrimp is indicative of the presence of potent cytotoxic components, which suggested that it might be used as antiproliferative and antitumor.
TABLE 1: BRINE SHRIMP LETHALITY DATA OF EXTRACTS AND ISOLATED COMPOUNDS OF CLEOME BRACHYCARPA
|Samples||LC50 (µg/ml, 24h)|
|Vincristine sulphate (Std.)||3.65||Compound-3||37.00|
CONCLUSION: The present study reported the isolation of a flavonol glycoside, a megastigmane glucoside, four methylated flavonoids, and two sesquiterpene oxides. Although, the brine shrimp lethality assay is rather inadequate regarding the elucidation of the mechanism of action, it is very useful to assess the bioactivity of the plant extracts. The results indicated for the first time that the aerial parts of Cleome brachycarpa extracts and the purified compounds exhibited significant cytotoxic activity (LC50 values < 100 μg/ml) using brine shrimp lethality assay and are considered as a source of natural agents that could be used as antiproliferative, antitumor and could provide leads to interesting pharmaceuticals of plant origin.
ACKNOWLEDGEMENT: The authors would like to acknowledge Dr. Mohamed El Gibaly for identification of plant material.
CONFLICT OF INTEREST: Nil
- Tackholm V: Students Flora of Egypt. Cairo University Cooperative printing Co., Egypt, Second Edition 1974: 167-69.
- Boulos L: Flora of Egypt, Azollaceae and Oxalidaceae, AL Hadara Publishing, Cairo, Egypt 1st Edition, 1999: 177-80.
- Sudhakar M, Rao CV, Rao PM and Raju DB: Evaluation of the antimicrobial activity of Cleome viscosa and Gmelina asiatica. Fitoterapia 2006; 77(1): 47-49.
- Mobiya AK, Ashutosh KPAG and Jeyakandan SM: Hepatoprotective effect of Cleome viscosa seeds in paracetamol induced hepatotoxic rats. International Journal of Pharmaceutical & Biological Archives 2010; 1(4): 399-03.
- Asis B, Biswakanth K, Pallab KH, Mazumder UK and Samit B: Evaluation of the anticancer activity of Cleome gynandra on Ehrlich's Ascites Carcinoma treated mice. Journal of Ethnopharmacology 2010; 129(1): 131-34.
- Mothana RAA, Mentel R, Reiss C and Lindequist U: Phytochemical screening and antiviral activity of some medicinal plants from the island Soqotra. Phytotherapy Research 2006; 20(4): 298-02.
- Meda NTR, Bangou MJ, Bakasso S, Millogo-Rasolodimby J and Nacoulma OG: Antioxidant activity of phenolic and flavonoid fractions of Cleome gynandra and Maerua angolensis of Burkina Faso. Journal of Applied Pharmaceutical Science 2013; 3(2): 36-42.
- Vishal TA, Mahamuni TJ and Karadge BA: Taxonomy and physiological studies in spider flower (Cleome species): a critical review. Plant Sciences Feed 2012; 2(3): 25- 46.
- Sharaf M, El-Ansari ME and Saleh NAM: Flavonoids of four Cleome and three Capparis species. Biochem Syst Eco 1997; 25(2): 161-66.
- Fushiya S, Kishi Y, Hattori K, Batkhuu J, Takano F, Singab ANB and Okuyama T: Flavonoids from Cleome droserifolia suppress NO production in activated macrophages in-vitro. Planta Med 1999; 65: 404-07.
- Ray AB, Chattopadhyay SK, Kumar S, Konno C, Kiso Y and Hikino H: Structures of cleomiscosins coumarino-lignoids of Cleome viscosa Tetrahedron 1985; 41(1): 209-14.
- Srivastava SK: Stigmasta-5, 24(28)-diene 3β-O-α-L-rhamnoside from Cleome viscose. Phytochemistry 1980; 19(11): 2510-11.
- Harraz FM, Ulubelen A, Osuz S and Tan N: Dammarane triterpenes from Cleome amblyocarpa. Phytochemistry 1995; 39(1): 175-78.
- Ahmad VU, Qazi S, Bin Zia N, Xu C and Clardy J: Cleocarpone, A triterpenoid from Cleome brachycarpa. Phyto-chemistry 1990; 29: 670.
- Viqar UA and Khisal A A: Deacetoxybrachycarpone, a trinortriterpenoid from Cleome brachycarpa. Phyto-chemistry 1986; 26(1): 315-16.
- Ahmad VU and Alvi KA: Deacetoxybrachycarpone a trinortriterpenoid from Cleome brachycarpa. Phyto-chemistry 1987; 26(1): 315-16.
- Hussein NS, Ahmed AA and Darwish FMK: Sesquiterpenes from Cleome droserifolia. Pharmazie 1994; 49(1): 76-77.
- El-Askary HI: Terpenoids from Cleome droserifolia (Forssk.) Del. Molecules 2005; 10: 971-77.
- Burke BA, Chan WR, Honkan VA, Blount JF and Manchand PS: Structure of cleomeolide an unusual bicyclic diterpene from Cleome viscose Tetrahedron 1980; 36(24): 3489-93.
- Rahman MA, Mossa JS, Al-Said MS and Al-Yahya MA: Medicinal plant diversity in the flora of Saudi Arabia, a report on seven plant families. Fitoterapi 2004; 75: 149-61.
- Qureshi R, Bhatti GR and Memon RA: Ethnonnedicinal uses of herbs from Northern Part of Nara Desert, Pakistan. Pak J Bot 2010; 42(2): 839-51.
- Muhammad AN and Salman AA: Anti-emetic activity of Cleome brachycarpa and Cleome viscosa in chicks. Universal Journal of Pharmacy 2013; 1(1): 96-99.
- Viqaruddin A and Khisal AA: Deacetoxybrachycarpone, a triterpenoid from Cleome brachycarpa. Phytochemistry 1987; 26(1): 315-16.
- Touchstone JC: Practice of Thin Layer Chromatography, Wiley-Interscience, New York, Third Edetion, 1992: 250-55.
- Meyer BN, Ferrigni NR, Putnam JE, Jacobsen JB, Nicholsand DE and Mclaughlin JL: Brine shrimp; a convenient general bioassay for active plant constituents. Planta Med 1982; 45(3): 1-34.
- Mc Lauglin JL, Chang CJ and Smith DL: Simple bench-top bioassays (brine shrimp and potato discs) for the discovery of plant antitumour compounds. In: Human Medicinal Agents from Plants Kinghorn AD and Balandrin MF (Eds.), ACS Symposium 534, American Chemical Society, Washington DC 1993: 112-37.
- Mabry TJ: Markham KR and Thomas MB: The Systematic Identification of Flavonoids, Springer Verlag, New York, 1970: 280.
- Tang Y, Lou F, Wang J and Zhuang S: Four new isoflavone triglycosides from Sophora japonica. J Nat Prod 2001; 64(8): 1107-10.
- Agrawal PK: C13-NMR of Flavonoids, Elsevier, Amesterdam, 1989.
- Otsuka H, Yao M, Kamada K and Takeda Y: Alangionosides G-M: glycosides of megastigmane derivatives from the leaves of Alangium premnifolium. Chem Pharm Bull 1995; 43: 754-59.
- Pabst A, Barron D, Sémon E and Schreier P: Two diastereomeric 3-oxo-a-ionol b-D-glucosides from raspberry fruit. Phytochemistry 1992; 31: 1649-52.
- Bhakuni DS, Joshi PP, Uprety H and Kapil RS: Roseoside a C13 glycoside from Vinca rosea. Phytochemistry 1974; 13: 2541-43.
- Ahmad VU, Zahid M, Ali MS, Jassbi AR, Abbas M, Ali Z and Iqbal MZ: Bucharioside and buchariol from Salvia bucharica. Phytochemistry 1999; 52: 1319-22.
- Bruno M, Torre MC, Rodríguez B and Omar AA: Guaiane sesquiterpenes from Teucrium leucocladum. Phyto-chemistry 1993; 34: 245-47.
How to cite this article:
Afifi MS: Phytochemical and biological investigation of Cleome brachycarpa Vahl. Growing in Egypt. Int J Pharm Sci & Res 2014; 5(9): 4008-14. doi: 10.13040/IJPSR.0975-8232.5(9).4008-14.
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