Establishment and Application of a High-throughput Drug Screening Assay Targeting Macrophage Migration Inhibitory Factor

doi:10.13560/j.cnki.biotech.bull.1985.2016.09.034

Abstract

Abstract: The aim is to establish a high-throughput drug screening assay targeting macrophage migration inhibitory factor（MIF）by UV-spectrophotometry. The target gene was molecularly cloned,prokaryotic expression system of Escherichia coli was used to purify,and then the highly-purified target protein was acquired. UV-spectrophotometry was utilized to establish the enzymatic system,in which then the conditions were optimized,and the proper high-throughput drug screening assay was set up and screened new MIF inhibitors from 384 small molecules. As results,the screening assay was established successfully,and 2 small molecules with high inhibitory rate were identified,while median inhibitory concentration（IC₅₀）was 59.07 μmol/L and 44.12 μmol/L, respectively. In conclusion,targeting MIF protein,the ideal high-throughput drug screening assay was established,which is appropriate for screening MIF inhibitors and conducive to the future drug development.

Key words: macrophage migration inhibitory factor, high-throughput screening, tautomerase activity, inhibitors

ZHANG Jing^{1, 2}, SUN Rui-qiu², TANG Yan-ting², LIU Xiang². Establishment and Application of a High-throughput Drug Screening Assay Targeting Macrophage Migration Inhibitory Factor[J]. Biotechnology Bulletin, 2016, 32(9): 253-259.

References

[1] Bloom B, Bennett B. Mechanism of a reaction in vitro associated with delayed-type hypersensitivity[J]. Science, 1966, 153（373）：80-82.
[2] Weiser W, Temple P, David J. Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor[J]. Proceedings of the National Academy of Sciences of the United States America, 1989, 86（19）：7522-7526.
[3] Merk M, Mitchell R, Endres S, et al. D-dopachrometautomerase（DDT or MIF-2）：doubling the MIF cytokine family[J]. Cytokine, 2012, 59（1）：10-17.
[4] Kim H, Kim K, Jung H, et al. Macrophage migration inhibitory factor enhances osteoclastogenesis through upregulation of RANKL expression from fibroblast-like synoviocytes in patients with rheumatoid arthritis[J]. Arthritis Research & Therapy, 2011, 13（43）：1-13.
[5] Calandra T, Bernhagen J, Bucala R. MIF as a glucocorticoid-induced modulator of cytokine production[J]. Nature, 1995, 377（6544）：68-71.
[6] Aeberli D, Yang Y, Mansell A, et al. Endogenous macrophage migration inhibitory factor modulates glucocorticoid sensitivity in macrophages via effects on MAP kinase phosphatase-1 and p38 MAP kinase[J]. FEBS Letters, 2006, 580（3）：974-981.
[7] Krockenberger M, Engel J, Kolb J, et al. Macrophage migration inhibitory factor expression in cervical cancer[J]. Journal of Cancer Research and Clinical Oncology, 2010, 136（5）：651-657.
[8] Bandres E, Bitarte N, Arias F, et al. microRNA-451 regulates macrophage migration inhibitory factor production and proliferation of gastrointestinal cancer cells[J]. Clinical Cancer Research, 2009, 15（7）：2281-2290.
[9] Guo, Y, Dai Y, Li W, et al. Expression and significance of macrophage migration inhibitory factor in bladder urothelial cell carcinoma[J]. Chinese Journal of Oncology, 2011（33）：28-31.
[10] Tan L, Ye X, Zhou Y, et al. Macrophage migration inhibitory factor is overexpressed in pancreatic cancer tissues and impairs insulin secretion function of beta-cell[J]. Journal of Translational medicine, 2014, 12：92.
[11] Razzaghi M, Mazloomfard M, Lotfi B. Macrophage migration inhibitory factor gene-173G>C polymorphism and risk of prostate cancer[J]. European Urology Supplements, 2012, 19（12）：142.
[12] Nabizadeh MM, Sima HR, Ghaffarzadehgan K, et al. Clinicopathol-ogical significance of macrophage migration inhibitory factor and its relation with p53 in gastric cancer[J]. Journal of Gastrointestinal Cancer, 2011, 42（1）：5-10.
[13] Santos L, Morand E. Macrophage migration inhibitory factor：a key cytokine in RA, SLE and atherosclerosis[J]. Clinicachimicaacta;International Journal of Clinical Chemistry, 2009, 399（1-2）：1-7.
[14] Sanchez-Niño M, Sanz A, Ihalmo P, et al. The MIF receptor CD74 in diabetic podocyte injury[J]. Journal of the American Society of Nephrology, 2009, 20（2）：353-362.
[15] Hawkins O, Verma B, Lightfoot S, et al. An HLA-presented fragment of macrophage migration inhibitory factor is a therapeutic target for invasivebreast cancer[J]. Journal of Immunology, 2011, 186（11）：6607-6616
[16] Grieb G, Merk M, Bernhagen J, et al. Macrophage migration inhibitory factor（MIF）：a promising biomarker[J]. Drug News Perspect, 2010, 23（4）：257-264.
[17] Vujicic M, Senerovic L, Nikolic I, et al. The critical role of macrop-hage migration inhibitory factor in insulin activity[J]. Cytokine, 2014, 69（1）：39-46.
[18] Sugimoto H, Taniguchi M, Nakagawa A, et al. Crystal structure of human D-dopachrometautomerase, a homologue of macrophage migration inhibitory factor, at 1. 54 Å resolution[J]. Biochemistry, 1999, 38（11）：3268-3279.
[19] Rosengren E, Aman P, Thelin S, et al. The macrophage migration inhibitory factor MIF is a phenylpyruvatetautomerase[J]. FEBS Letters, 1997, 417（1）：85-88.
[20] Swope M, Sun H, Blake P, et al. Direct link between cytokine activity and a catalytic site for macrophage migration inhibitory factor[J]. The EMBO Journal, 1998, 17（13）：3534 - 3541.
[21] Xu L, Li Y, Sun H, et al. Current developments of macrophage migration inhibitory factor（MIF）inhibitors[J]. Drug Discovery Today, 2013, 18（11-12）：592-600.
[22] Hajer O, Liu M, Farah E, et al. Kinetic-based high-throughput screening assay to discover novel classes of macrophage migration inhibitory factor inhibitors[J]. Journal of Biomolecular Screening, 2010, 15（4）：347-358.
[23] Zierow S, Crichlow G, Lolis E. A novel, macrophage migration inhibitory factor suicide substrate inhibits motility and growth of lung cancer cells[J]. Cancer Research, 2008, 68：7253-7257.
[24] Xu L, Zhang Y, Zheng L, et al. Discovery of novel inhibitors targeting the macrophage migration inhibitory factor via structure-based virtual screening and bioassays[J]. Journal of Medicinal Chemistry, 2014, 57：3737-3745.