In-vitro immunomodulatory and anti-cancerous activities of biotransformed products of Dianabol through Azadirachta indica and its molecular docking studies

Chemistry Central Journal

Saifullah³  Saifullah Khan³  Azizuddin²  Sobia Ahsan Halim¹  Muhammad Kashif³  Almas Jabeen¹  Muhmammad Asif¹  Muhammad Ahmed Mesaik¹  Zaheer Ul-Haq¹  Ahsana Dar¹  Muhammad Iqbal Choudhary¹ 

[1] Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan

[2] Department of Chemistry, Federal Urdu University for Arts, Science & Technology, Gulshan-e-Iqbal Campus, Karachi 75300, Pakistan

[3] Department of Biotechnology, Faculty of Science, University of Karachi, Karachi 75270, Pakistan

关键词: Molecular docking studies;    Anti-cancer activity;    Anti-inflammatory activity;    Biotransformation;   

Others  :  787831
DOI  :  10.1186/1752-153X-7-163

 received in 2013-02-28, accepted in 2013-06-06,  发布年份 2013

【 摘 要 】

Background

Plant Biotransformation is one of the tools for structural modifications of the organic substrate of low, moderate or high biological value utilizing plant cultured cells, these modifications of organic structures may lead to biologically augmented products and which may be ultimately substantial in cure or improvement of various morbidities and diseases.

Results

Azadirachta indica A. Juss. suspension culture was employed for the biotransformation of dianabol (1) for the first time, and two metabolites, 17β-hydroxy-17α-methyl-5α-androst-1-en-3-one (2), and 17β-hydroxy-17α-methyl-5α-androstan-3-one (3) were obtained.

Conclusions

Most important aspect of this work was the evaluation of metabolite 2, which strongly and differentially suppressed [not affecting whole blood and human polymorphonuclear cells (PMN)] the phytohemagglutinin (PHA)-activated T-cell proliferation (IC₅₀: <10.33 μM), and also found to inhibit IL-2 production (IC₅₀: 16.89 ± 1.32) unlike metabolite 3 and compound 1. Compound 2 also exhibited anticancer activity against lung cancer cell line; NCI-H460, it moderately inhibited the growth of cancer cells (22.5 ± 4.15 μM). Furthermore, a good correlation between the predicted binding energies of the compounds acquired by the FlexX program and the experimental affinities were speculated upon interacting with IL-2 protein during molecular docking studies.

【 授权许可】

   
2013 Saifullah et al.; licensee Chemistry Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Scheme 1

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Figure 5.

【 参考文献 】

• [1]Charlwood BV, Rhodes MJC: Secondary Products from Plant Tissue Cultures. Oxford: Clarendon Press; 1990.
• [2]Lowe KC, Davey MR, Power JB: Plant tissue culture: past, present and future. Plant Tiss Cult Biotechnol 1996, 2:175-186.
• [3]Charlwood BV, Hegarty PK, Charlwood KA, Lippin G, Tampion J, Stride J: The synthesis and Biotransformation of monoterpenes by Plant Cells in Culture. In Secondary Metabolism in Plant Cell Cultures. Edited by Morris P, Scragg AH, Stafford A, Fowler MW. London: Cambridge University Press; 1986:113-116.
• [4]Dia J, Guo H, Lu D, Zhu W, Zhang D, Zheng J, Guo D: Biotransformation of 2α, 5α, 10α, 14α, tetra acetoxy –(20), 11 taxadiene by Ginkgo cell suspension cultures. Tetrahedron Lett 2001, 42:4677-4679.
• [5]Reihard E, Alfermann AW: Advances in biochemical engineering. New York: Springer; 1980.
• [6]Suga T, Hirata T: Biotransformation of exogenous substrates by plant cell cultures. Phytochemistry 1990, 29:2393-2406.
• [7]Ishihara K, Hamada H, Hirata H, Nakajima N: Biotransformation using plant cultured cells. J Mol Catal B: Enzym 2003, 23:145-170.
• [8]Mukaiyama T, Yamada T, Nagata T, Imagawa K: Asymmetric aerobic epoxidation of unfunctionalized olefins catalyzed by optically active α-alkoxycarbonyl-β-ketoiminato manganese (III) complexes. Chem Lett 1993, 2:327-330.
• [9]Sharpless KB, Amberg W, Bennani YL, Crispino GA, Hartung J, Jeong K, Kwon H, Morikawa K, Wang Z, Xu D, Zhang X: The osmium-catalyzed asymmetric dihydroxylation: a new ligand class and a process improvement. J Org Chem 1992, 57:2768-2771.
• [10]Sakamaki H, Take M, Matsumoto T, Iwadare T, Ichinohe Y: Transformation of cycloartanyl acetate into B-homo triterpenoids. J Org Chem 1988, 53:2622-2624.
• [11]Sakamaki H, Itoha KI, Taniai T, Kitanaka S, Takagi Y, Chai W, Horiuchi CA: Biotransformation of valencene by cultured cells of Gynostemma pentaphyllum. J Mol Cat B Enzym 2005, 32:103-106.
• [12]Choudhary MI, Siddiqui ZA, Khan S, Saifullah , Musharraf SG, Atta-ur-Rahman : Biotransformation of (−)-Caryophyllene Oxide by cell Suspension culture of Catharanthus roseus. Z Naturforsch 2006, 61b:197-200.
• [13]Azizuddin , Saifullah , Khan S, Choudhary MI, Atta-Ur-Rahman : Biotransformation of dydrogesterone by cell suspension cultures of Azadirachta indica. Turk J Chem 2008, 32:141-146.
• [14]Zafar S, Yousuf S, Kayani HA, Saifullah KS, Al-Majid AM, Choudhary MI: Biotransformation of oral contraceptive ethynodiol diacetate with microbial and plant cell cultures. [http://dx.doi:10.1186/1752-153X-6-109] webciteChem Cent J 2012.
• [15]Musharraf SG, Jalaluddin , Akhter M, Parvez M, Saifullah , Khan S, Yousaf S, Khan S, Choudhary MI: Biotransformation of an antimalarial drug, artemether by plant and fungal cell cultures. J Mol Cat B Enzym 2012, 82:80-83.
• [16]Biswas K, Chattopadhyay I, Banerjee RK, Bandyopadhyay U: Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci 2002, 82:1336-1345.
• [17]Páduaa RM, Meitinger N, Filho JDS, Waibel R, Gmeiner P, Braga FC, Kreis W: Biotransformation of 21-O-acetyl-deoxycorticosterone by cell suspension cultures of Digitalis lanata (strain W.1.4). Steroids 2012, 77:1373-1380.
• [18]Saifullah , Khan S, Azizuddin , Kashif A, Dar A, Choudhary MI: Anti-cancerous compound. 2013. U.S. Patent No. 18025 A1, http://www.faqs.org/patents/app/20130018025 webcite
• [19]Saifullah , Khan S, Mesaik MA, Zaheer-ul-Haq , Jabeen A, Halim SA, Choudhary MI: Anti-inflammtory compound. 2012. U.S. Patent No. 8,329,678 B1, http://www.freepatentsonline.com/8329678.html webcite
• [20]Halim SA, Abdalla OM, Mesaik MA, Wadood A, Zaheer-il-Haq , Kontoyianni M: Identification of novel Interleukin-2 inhibitors through computational approaches. Mol Diver 2013, 17:345-355.
• [21]Mesaik MA, Zaheer-ul-Haq , Murad S, Ismail Z, Abdullah NR, Gill HK, Atta-Ur-Rahman , Yousaf M, Siddiqui RA, Ahmad A, Choudhary MI: Biological and molecular docking studies on coagulin-H: Human IL-2 novel natural inhibitor. Mol Immunol 2006, 43:1855-1863.
• [22]Hampel VB, Kraemer JM: Die Kernresonanzspektren von steroiden in polaren lösungsmitten II. Tetrahedron 1966, 22:1601-1613.
• [23]Parr MK, Blatt C, Zierau O, Hess C, Gutschow M, Fussholler G, Opfermann G, Schanzer W, Diel P: Endocrine characterization of the designer steroid methyl-1-testosterone: investigations on tissue-specific anabolic-androgenic potency, side effects, and metabolism. Endocrinology 2011, 152:4718-4728.
• [24]Gómez C, Pozo OJ, Marcos J, Segura J, Ventura R: Alternative long-term markers for the detection of methyltestosterone misuse. Steroids 2013, 78:44-52.
• [25]17α-Methyl-1-testosterone. http://www.lookchem.com/cas-65/65-04-3.html webcite
• [26]Thevis M, Schänzer WJ: Mass spectrometric analysis of androstan-17β-ol-3-one and androstandiene-17β-ol-3-one isomers. Am Chem Soc Mass Spect 2005, 16:1660-1669.
• [27]Takata N, Shiraki K, Takano R, Hayashi Y, Terada K: Cocrystal screening of stanolone and mestanolone using slurry crystallization. Cry Gro Des 2008, 8:3032-3037.
• [28]Dihydrotestosterone. http://ctdbase.org/detail.go?type=chem&acc=D013196 webcite
• [29]Chhikara BS, Chandra R, Tandon V: IBX in an ionic liquid: eco-friendly oxidation of 17α-methylandrostan-3β, 17β-diol, an intermediate in the synthesis of anabolic oxandrolone. Tetraheron Lett 2004, 45:7585-7588.
• [30]Murashige T, Skoog F: A revised medium for rapid growth and biossays with tobacco tissue cultures. Physiol Plant 1962, 15:473-497.
• [31]Helfand S, Werkmeister J, Roder J: Chemiluminescence response of human natural killer cells. The relationship between target cell binding, chemiluminescence, and cytolysis. J Exp Med 1982, 156:492-505.
• [32]Haklar G, Ozveri ES, Yuksel M, Aktan A, Yalynn AS: Different kinds of reactive oxygen and nitrogen species were detected in colon and breast tumors. Cancer Lett 2001, 165:219-224.
• [33]Nielsen M, Gerwien J, Nielsen M, Geisler C, Ropke C, Odum N: MHC class II ligation induces CD58 (LFA-3)-mediated adhesion in human T cells. Exp Clin Immunogenet 1998, 15:61-68.
• [34]Boyum A: Isolation of mononuclear cells and granulocytes from human blood. Isolation of mononuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation. Scand J Clin Lab Invest 1968, 21:77-89.
• [35]Skehan P, Streng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR: New colorimetric cytotoxicity assay for anti-cancer drug screening. J Nat Cancer Inst 1990, 82:1107-1112.
• [36]Tilley JW, Chen L, Fry DC, Emerson SD, Powers GD, Biondi D, Varnell T, Trilles R, Guthrie R, Mennona F, Kaplan G, LeMahieu RA, Carson M, Han R-J, Liu CM, Palermo R, Ju G: Identification of a small molecule inhibitor of the IL-2/IL-2Rr receptor interaction which binds to IL-2. J Am Chem Soc 1997, 119:7589-7590.
• [37]Arkin Arkin MR, Randal M, DeLano WL, Hyde J, Luong TN, Oslob JD, Raphael D, Taylor L, Wang J, McDowell RS, Wells JA, Braisted AC: Binding of small molecules to an adaptive protein–protein interface. PNAS 2003, 100:1603-1608.
• [38]SYBYL molecular modeling software. St. Louis MO: Tripos Associated Ltd;
• [39]TRIPOS Inc 1699 South Hanley Road, St. Louis, MO 63144, USA;
• [40]Rarey M, Kramer B, Lengauer T, Klebe G: A fast flexible docking method using an incremental construction algorithm. J Mol Biol 1996, 261:470-489.
• [41]Theoretical and computational Biophysics Group: Visual Molecular Dynamics Version 1.8.6. 405 N. Matthews, Urbana, IL 61801: University of Illinois & Beckman Institute;