| Abstract: |
I - Compounds isolated from Acanthodendrilla sp.:The freeze dried sponge was powdered and extracted with methanol then with acetone, which were combined together. The total extract was subjected to liquid liquid extraction, where n-hexane, ethylacetate, n-butanol and aqueous fractions were obtained.Ten milligram of each fraction were used for preliminary bilogical screening, through brine shrimp assay and antimcrobial activity. According to these results, the n-hexane fraction was selected to be chemically investigated to identify the metabolites of this active fraction, where 8 compounds were isolated (Table 26).Biological Investigation of the Isolated CompoundsI-Antimicrobial Activity:As the hexane fraction from which compounds 1-8 were isolated showed antimicrobial activity, the pure compounds were tested to investigate this activity.It was fouund that luffariellolide (1), methyl- luffariellolide (2), ethyl- luffariellolide (3) and acantholide-C (6), have antimicrobial activity. Luffariellolide was the most active against the test bacterial strains (B. subtilis, E. coli and S. aureus) in concentrations of 5 and 10 µg, while it experessed antifungal activity in the concentration of 10µg only, where it showed mild antimycotic ction against C. albicans and strong fungicidal activity against C. herbarum that was evaluated with MC50 of 10.4µg/ml.Concerning the other compounds:• Methyl-luffariellolide showed less activity aginst B. subtilis in both concentrations, against S. aureus it was active in concentration of 10µg and it was comparable to luffariellolide, inactive against E. coli in both concentrations. It has antimycotic acitvity comparable to luffariellolide and fungistatic activity against C. herbarum.• Ethyl-luffariellolide has antimycotic acitvity comparable to luffariellolide, but no activity against C. herbarum nor S. aureus . Against E. coli it showed higher activity than luffariellolide in the concentration of 10µg while against B. subtilis it showed less activity than luffariellolide in the concentration of 10µg and equall activity in the concentration of 5µg.• Acantholide-C showed higher activity than luffariellolide against E. coli in a concentration of 10µg, it was less fungistatic against C. herbarum than methyl-luffariellolide, against B. subtilis and S. aureus it shwoed activity in the concentration of 10µg only and it was less than luffariellolide.II- Cytotoxicity:The cytotoxic effect was tested against L5178Y, PC12 and Hela cell lines, it was found that luffariellolide (1) and acantholide-D (7) were the only active compounds, and the activity was against L5187Y cell line only (mouse lymphoma).• Luffariellolide, gave 100% growth supression at a concentration of 10µg/ml and by 3µg/ml gave supression of 42%, the ED50was calculated as 3.3µg/ml. To the best of my knowledge this was the first evaluation of the cytotoxic activity for luffariellolide.• Acantholide-D, gave 82.5% growth suppression at concentration of 10µg/ml, but at 3µg/ml there were no growth inhibition, the ED50 was calculated as 7µg/ml.II - Compounds isolated from Stylissa flabelliformis:The freeze dried sponge was powdered and extracted with methanol then with acetone, which were combined together. The total extract was subjected to liquid liquid extraction, where n-hexane, ethylacetate, n-butanol and aqueous fractions were obtained.Ten milligram of each fraction were used for preliminary bilogical screening, through fish feeding assay and antimcrobial activity. The fractions showed activity only with the fish feeding assay, which showed that the ethyl acetate gave 90% deterrence activity. According to this result the ethyl acetate fraction was selected to be chemically investigated, where 6 compounds were isolated (table 27).Biological Invetigation of the Isolated CompoundsI-Cytotoxiciy:The cytotoxicity of the isolated compounds was tested against; Hela, L5178Y and PC12 cell lines.It was found that hymnialdisin (13) showed 100% growth inhbition of the Hela and L5178Y cell lines, while it was inactive against PC12 cell line. Dibromocantharillin (14) showed moderate growth inhibition against the L5178Y cell line without any activity on the other cell lines. 2-Bromoaldisin (11) showed mild growth inhibition against L5178Y cell line without any activity on the other cell lines. The other compounds 9, 10 and 12 were completely inactive against all the used cell lines.II-Antimicrobial activity:The antimicrobial activity of the isolated compounds was also tested. It was found that non of the compounds has any antibacterial activity against the used strians (B. subtilis, E. coli and S. aureus).The antifungal activity was tested aginst C. herbarum and C. cucumerinum, dibromocantharillin was the only active compound, as it gave fungicidal effect expressed by zone of inhbition of 16 mm.III-Compounds isolated from Pericarax heteroraphis:The freeze dried sponge was powdered and extracted with methanol then with acetone, which were combined together. The total extract was subjected to liquid liquid extraction, where n-hexane, ethylacetate, n-butanol and aqueous fractions were obtained.Ten milligram of each fraction were used for preliminary bilogical screening, through fish feeding assay and antimcrobial activity. The fractions showed activity only with the fish feeding assay, which showed that the ethyl acetate gave 78% deterrence activity. According to this result the ethyl acetate fraction was selected to be chemically investigated, where 3 compounds were isolated (table 28).IV-Compounds isolated from Haliclona sp. :The freeze dried sponge was powdered and extracted with methanol then with acetone, which were combined together. The total extract was subjected to liquid liquid extraction, where n-hexane, ethylacetate, n-butanol and aqueous fractions were obtained.The EtoAc fraction was subjected to HPLC preparative where compound 18 and 19 were isolated. The n-hexane fraction was subjected for silica gel column chromatography, eluted with hex./EtoAc gradient, where compound 20 was isolated, the structure of the isolated compounds were listed in table 29.The Indolecarboxaldehyd (18) was found to has mild cytotoxic activity (47%) as it inhibited the growth of L5187Y cell line.
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