选择性雌激素受体调节剂的筛选及其活性评价
|
摘要
雌激素受体(Estrogen Receptor, ER)是核受体超家族的成员之一,是一类重要的转录调控因子,是治疗乳腺癌、骨质疏松等疾病的重要药物作用靶标。选择性雌激素受体调节剂在治疗ER相关疾病上具有很高的临床应用价值。我们通过计算机虚拟筛选和生物实验相结合的手段来寻找新的选择性ER配体。
首先我们采用分子动力学结合虚拟筛选的策略,从商业数据库中搜寻选择性ER配体。通过分子动力学模拟得到ER晶体结构的平均平衡构象作为虚拟筛选的结构基础,然后进行三轮基于分子对接的虚拟筛选,最后确定了70个候选化合物。在活性测试中,我们利用ER与其共激活因子的相互作用构建了酵母双杂交系统,通过检测报告基因活性的方式评价待测化合物的活性和作用类型。结果发现,在这70个化合物中至少有18个表现出良好的ER配体活性。其中有两个双效配体：同时表现为ERa拮抗剂及ERp激动剂；还有11个ER激动剂和5个ER拮抗剂,且不少化合物表现出较高的ER亚型选择性。
通过细胞转染和报告基因检测实验,我们验证了这些化合物在细胞水平上能够改变ER的转录激活活性。MTT实验表明,具有ER拮抗活性的化合物能够抑制乳腺癌细胞增殖。我们还较深入地探讨了ER拮抗剂JB042对乳腺癌细胞的生长抑制作用。通过western blot和流式细胞周期分析实验,发现JB042可以下调雌二醇诱导的雌激素效应基因cyclin D1的表达,并将细胞周期阻滞于G1期,最终抑制癌细胞增殖。
Estrogen receptor (ER), which belongs to the nuclear receptor super family, is a kind of important transcriptional regulator and is an essential drug target involving in breast cancer and osteoporosis. More and more research showed that selective estrogen receptor modulators had higher value in clinical application. We use computer virtual screening (VS) and bioassay to discover new selective ER ligands.
First, we employed a strategy that combined molecular dynamics (MD) simulation with virtual screening to discover novel ER ligands from a commercial database. The average structure taken from equilibrated stage of ER crystal structure by MD simulation was prepared as the structural basis, and three rounds of VS were subsequently performed. Then 70 candidate compounds were selected for bioassay. We constructed yeast two-hybrid system to screen ligands and determine the capacity of compounds, which was based on the interaction of ER and its activator. As a result, more than 18 potent ligands were discovered. Among them, dual profile was observed in two ligands, as antagonists for ERαand agonists for ERβ; 11 agonists and 5 antagonists were discovered as well, and some of them showed high ER subtype selectivity.
We performed cell transfection and luciferase activity assays and MTT assays to check the role of the novel ligands on ER transcriptional activation and antiproliferative potencies on breast cancer cell line. The novel ligands could effects ER transcription and the compounds that showed antagonistic activities on ERβor ERαdisplayed certain antiproliferative activities. We dissected the action of ER antagonist JB042 on suppressing breast caner cells proliferation. By western blot and flow cytometry analysis, we found that treatment with JB042 could down-regualte the expression of cycline D1 indued by estrogen and suppress the progression of G1 to S, subsequently inhibited cell proliferation.
首先我们采用分子动力学结合虚拟筛选的策略,从商业数据库中搜寻选择性ER配体。通过分子动力学模拟得到ER晶体结构的平均平衡构象作为虚拟筛选的结构基础,然后进行三轮基于分子对接的虚拟筛选,最后确定了70个候选化合物。在活性测试中,我们利用ER与其共激活因子的相互作用构建了酵母双杂交系统,通过检测报告基因活性的方式评价待测化合物的活性和作用类型。结果发现,在这70个化合物中至少有18个表现出良好的ER配体活性。其中有两个双效配体：同时表现为ERa拮抗剂及ERp激动剂；还有11个ER激动剂和5个ER拮抗剂,且不少化合物表现出较高的ER亚型选择性。
通过细胞转染和报告基因检测实验,我们验证了这些化合物在细胞水平上能够改变ER的转录激活活性。MTT实验表明,具有ER拮抗活性的化合物能够抑制乳腺癌细胞增殖。我们还较深入地探讨了ER拮抗剂JB042对乳腺癌细胞的生长抑制作用。通过western blot和流式细胞周期分析实验,发现JB042可以下调雌二醇诱导的雌激素效应基因cyclin D1的表达,并将细胞周期阻滞于G1期,最终抑制癌细胞增殖。
Estrogen receptor (ER), which belongs to the nuclear receptor super family, is a kind of important transcriptional regulator and is an essential drug target involving in breast cancer and osteoporosis. More and more research showed that selective estrogen receptor modulators had higher value in clinical application. We use computer virtual screening (VS) and bioassay to discover new selective ER ligands.
First, we employed a strategy that combined molecular dynamics (MD) simulation with virtual screening to discover novel ER ligands from a commercial database. The average structure taken from equilibrated stage of ER crystal structure by MD simulation was prepared as the structural basis, and three rounds of VS were subsequently performed. Then 70 candidate compounds were selected for bioassay. We constructed yeast two-hybrid system to screen ligands and determine the capacity of compounds, which was based on the interaction of ER and its activator. As a result, more than 18 potent ligands were discovered. Among them, dual profile was observed in two ligands, as antagonists for ERαand agonists for ERβ; 11 agonists and 5 antagonists were discovered as well, and some of them showed high ER subtype selectivity.
We performed cell transfection and luciferase activity assays and MTT assays to check the role of the novel ligands on ER transcriptional activation and antiproliferative potencies on breast cancer cell line. The novel ligands could effects ER transcription and the compounds that showed antagonistic activities on ERβor ERαdisplayed certain antiproliferative activities. We dissected the action of ER antagonist JB042 on suppressing breast caner cells proliferation. By western blot and flow cytometry analysis, we found that treatment with JB042 could down-regualte the expression of cycline D1 indued by estrogen and suppress the progression of G1 to S, subsequently inhibited cell proliferation.
引文
[1]Jensen EV, Joconbson HI. Basic guides to the mechanism of estrogen action. Recent Prog Horm Res.1962,18 (2):387-414
[2]Greene GL, Paul G, Michael W, Andrew B, Yvonne H, John S. Sequence and expression of human estrogen receptor complementary DNA. Science.1986,231 (4742): 1150-1154
[3]Green S, Walter P, Kumar V, Krust A, Bornert J-M, Argos P, et al. Human oestrogen receptor cDNA:sequence, expression and homology to v-erb-A. Nature.1986,320 (6058):134-139
[4]Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA. Cloning of a novel receptor expressed in rat prostate and ovary. Proceedings of the National Academy of Sciences of the United States of America.1996,93 (12):5925-5930
[5]Enmark E, Pelto-Huikko M, Grandien K, Lagercrantz S, Lagercrantz J, Fried G, et al. Human estrogen receptor β-Gene structure, chromosomal localization, and expression pattern. J Clin Endocrinol Metab.1997,82 (12):4258-4265
[6]Kong EH, Pike ACW, Hubbard RE. Structure and mechanism of the oestrogen receptor. Biochem Soc Trans.2003,31:56-59
[7]Mosselman S, Polman J, Dijkema R. ER β:Identification and characterization of a novel human estrogen receptor. FEBS Letters.1996,392 (1):49-53
[8]Ogawa S, Inoue S, Watanabe T, Hiroi H, Orimo A, Hosoi T, et al. The complete primary structure of human estrogen receptor beta (hER beta) and its heterodimerization with ER alpha in vivo and in vitro. Biochemical and Biophysical Research Communications. 1998,243(1):122-126
[9]Zhao L, O'Neill K, Diaz Brinton R. Selective estrogen receptor modulators (SERMs) for the brain:Current status and remaining challenges for developing NeuroSERMs. Brain Research Reviews.2005,49 (3):472-493
[10]Kumar V, Green S, Stack G, Berry M, Jin J-R, Chambon P. Functional domains of the human estrogen receptor. Cell.1987,51 (6):941-951
[11]Tora L, White J, Brou C, Tasset D, Webster N, Scheer E, et al. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell.1989, 59 (3):477-487
[12]Berry M, Metzager D, Chambon P. Role of the two activating domains of the oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the anti-oestrogen 4-hydroxytamoxifen. The EMBO Journal.1990,9 (9):2811-2818
[13]Tzukerman M, Esty A, Santiso-Mere D, Danielian P, Parker M, Stein R, et al. Human estrogen receptor transactivational capacity is determined by both cellular and promoter context and mediated by two functionally distinct intramolecular regions. Mol Endocrinol.1994,8 (1):21-30
[14]Fawell SE, White R, Hoare S, Sydenham M, Page M, Parker MG Inhibition of estrogen receptor-DNA binding by the "pure" antiestrogen ICI 164,384 appears to be mediated by impaired receptor dimerization. Proceedings of the National Academy of Sciences of the United States of America.1990,87 (17):6883-6887
[15]Dauvois S, Danielian PS, White R, Parker MG. Antiestrogen ICI 164,384 reduces cellular estrogen receptor content by increasing its turnover. Proceedings of the National Academy of Sciences of the United States of America.1992,89 (9): 4037-4041
[16]Kuiper GGJM, Carlsson B, Grandien K, Enmark E, Haggblad J, Nilsson S, et al. Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology.1997,138 (3):863-870
[17]Pike ACW, Brzozowski AM, Hubbard RE. A structural biologist's view of the oestrogen receptor. The Journal of Steroid Biochemistry and Molecular Biology.2000,74 (5): 261-268
[18]Shiau AK, Barstad D, Loria PM, Cheng L, Kushner PJ, Agard DA, et al. The structural basis of estrogen receptor/coactivator recognition and the antagonism of this interaction by Tamoxifen. Cell.1998,95 (7):927-937
[19]Pike ACW, Brzozowski AM, Hubbard RE, Bonn T, Thorsell A-G, Engstrom O, et al. Structure of the ligand-binding domain of oestrogen receptor beta in the presence of a partial agonist and a full antagonist. EMBO J.1999,18 (17):4608-4618
[20]Pike ACW, Brzozowski AM, Walton J, Hubbard RE, Thorsell A-G, Li Y-L, et al. Structural Insights into the Mode of Action of a Pure Antiestrogen. Structure.2001,9 (2):145-153
[21]McKenna NJ, Lanz RB, O'Malley BW Nuclear receptor coregulators:cellular and molecular biology. Endocr Rev.1999,20 (3):321-344
[22]Klinge CM. Estrogen receptor interaction with co-activators and co-repressors. Steroids. 2000,65 (5):227-251
[23]Tremblay G, Giguere V. Coregualtors of estrogen receptor action. Critical Reviews in Eukaryotic Gene Expression.2002,12 (1):1-22
[24]Heery DM, Kalkhoven E, Hoare S, Parker MG. A signature motif in transcriptional co-activators mediates binding to nuclear receptors. Nature.1997,387 (6634):733-736
[25]Lannigan DA. Estrogen receptor phosphorylation. Steroids.2003,68 (1):1-9
[26]Kato S. Estrogen receptor-mediated cross-talk with growth factor signaling pathways. Breast Cancer.2001,8 (1):3-9
[27]Kousteni S, Bellido T, Plotkin LI, O'Brien CA, Bodenner DL, Han L, et al. Nongenotropic, Sex-Nonspecific Signaling through the Estrogen or Androgen Receptors:Dissociation from Transcriptional Activity. Cell.2001,104 (5):719-730
[28]Nemere I, Pietras RJ, Blackmore PF. Membrane receptors for steroid hormones:Signal transduction and physiological significance. Journal of Cellular Biochemistry.2003,88 (3):438-445
[29]Wang Z, Zhang X, Shen P, Loggie BW, Chang Y, Deuel TF. A variant of estrogen receptor-α, hER-a36:Transduction of estrogen- and antiestrogen-dependent membrane-initiated mitogenic signaling. Proceedings of the National Academy of Sciences.2006,103 (24):9063-9068
[30]Funakoshi T, Yanai A, Shinoda K, Kawano MM, Mizukami Y. G protein-coupled receptor 30 is an estrogen receptor in the plasma membrane. Biochemical and Biophysical Research Communications.2006,346 (3):904-910
[31]Okamoto T, Schlegel A, Scherer PE, Lisanti MP. Caveolins, a Family of Scaffolding Proteins for Organizing reassembled Signaling Complexes at the Plasma Membrane. Journal of Biological Chemistry.1998,273 (10):5419-5422
[32]Guo X, Razandi M, Pedram A, Kassab G, Levin ER. Estrogen Induces Vascular Wall Dilation. Journal of Biological Chemistry.2005,280 (20):19704-19710
[33]Lu Q, Pallas DC, Surks HK, Baur WE, Mendelsohn ME, Karas RH. Striatin assembles a membrane signaling complex necessary for rapid, nongenomic activation of endothelial NO synthase by estrogen receptor. Proceedings of the National Academy of Sciences of the United States of America.2004,101 (49):17126-17131
[34]Castoria G, Migliaccio A, Bilancio A, Di Domenico M, de Falco A, Lombardi M, et al. PI3-kinase in concert with Src promotes the S-phase entry of oestradiol-stimulated MCF-7 cells. EMBO J.2001,20 (21):6050-6059
[35]Fernando RI, Wimalasena J. Estradiol Abrogates Apoptosis in Breast Cancer Cells through Inactivation of BAD:Ras-dependent Nongenomic Pathways Requiring Signaling through ERK and Akt. Mol Biol Cell.2004,15 (7):3266-3284
[36]Weihua Z, Warner M, Gustafsson J-A. Estrogen receptor beta in the prostate. Molecular and Cellular Endocrinology.2002,193 (1-2):1-5
[37]Telleria CM, Zhong L, Deb S, Srivastava RK, Park KS, Sugino N, et al. Differential Expression of the Estrogen Receptors alpha and beta in the Rat Corpus Luteum of Pregnancy:Regulation by Prolactin and Placental Lactogens. Endocrinology.1998,139 (5):2432-2442
[38]Iafrati MD, Karas RH, Aronovitz M, Kim S, Sullivan TR, Lubahn DB, et al. Estrogen inhibits the vascular injury response in estrogen receptor α-deficient mice. Nat Med. 1997,3 (5):545-548
[39]Karas RH, Schulten H, Pare G, Aronovitz MJ, Ohlsson C, Gustafsson J-A, et al. Effects of Estrogen on the Vascular Injury Response in Estrogen Receptor alpha, beta (Double) Knockout Mice. Circ Res.2001,89 (6):534-539
[40]Leav I, Lau K-M, Adams JY, McNeal JE, Taplin M-E, Wang J, et al. Comparative Studies of the Estrogen Receptors beta and alpha and the Androgen Receptor in Normal Human Prostate Glands, Dysplasia, and in Primary and Metastatic Carcinoma. Am J Pathol.2001,159(1):79-92
[41]Peters RK, Pike MC, Chang WWL, Mack TM. Reproductive factors and colon cancers. Br J Cancer.1990,61 (5):741-748
[42]Wenbin L. Age, sex and the risk of grade-specific second primary colorectal cancer: Evidence for the protective effect of female hormone. European Journal of Cancer. 2007,43 (12):1856-1861
[43]Slattery ML, Sweeney C, Murtaugh M, Ma KN, Wolff RK, Potter JD, et al. Associations between ERα, ERβ, and AR Genotypes and Colon and Rectal Cancer. Cancer Epidemiology Biomarkers & Prevention.2005,14 (12):2936-2942
[44]Foley EF, Jazaeri AA, Shupnik MA, Jazaeri O, Rice LW. Selective Loss of Estrogen Receptor β in Malignant Human Colon. Cancer Research.2000,60 (2):245-248
[45]Campbell-Thompson M, Lynch IJ, Bhardwaj B. Expression of Estrogen Receptor (ER) Subtypes and ER β Isoforms in Colon Cancer. Cancer Research.2001,61 (2):632-640
[46]Konstantinopoulos PA, Kominea A, Vandoros G, Sykiotis GP, Andricopoulos P, Varakis I, et al. Oestrogen receptor beta (ER beta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour's dedifferentiation. European Journal of Cancer.2003,39 (9):1251-1258
[47]Castiglione F, Taddei A, Degl Innocenti DR, Buccoliero AM, Bechi P, Garbini F, et al. Expression of Estrogen Receptor β in Colon Cancer Progression. Diagnostic Molecular Pathology.2008,17 (4):231-236
[48]Wada-Hiraike O, Warner M, Gustafsson J-A. New developments in oestrogen signalling in colonic epithelium. Biochem Soc Trans.2006,34 (6):1114-1116
[49]Martineti V, Picariello L, Tognarini I, Sala SC, Gozzini A, Azzari C, et al. ERβ is a potent inhibitor of cell proliferation in the HCT8 human colon cancer cell line through regulation of cell cycle components. Endocr Relat Cancer.2005,12 (2):455-469
[50]Fiorelli G, Picariello L, Martineti V, Tognarini I, Tonelli F, Brandi ML. Estrogen metabolism in human colorectal cancer cells. The Journal of Steroid Biochemistry and Molecular Biology.2002,81 (3):281-289
[51]Di Domenico M, Castoria G, Bilancio A, Migliaccio A, Auricchio F. Estradiol Activation of Human Colon Carcinoma-derived Caco-2 Cell Growth. Cancer Research. 1996,56 (19):4516-4521
[52]Sims NA, Dupont S, Krust A, Clement-Lacroix P, Minet D, Resche-Rigon M, et al. Deletion of estrogen receptors reveals a regulatory role for estrogen receptors-β in bone remodeling in females but not in males. Bone.2002,30 (1):18-25
[53]Lindberg MK, Moverare S, Skrtic S, Gao H, Dahlman-Wright K, Gustafsson J-A, et al. Estrogen Receptor (ER)-β Reduces ER α-Regulated Gene Transcription, Supporting a "Ying Yang" Relationship between ER α and ER β in Mice. Mol Endocrinol.2003,17 (2):203-208
[54]Persson I, Weiderpass E, Bergkvist L, Bergstrom R, Schairer C. Risks of breast and endometrial cancer after estrogen and estrogen-progestin replacement. Cancer Causes and Control.1999,10 (4):253-260
[55]Katzenellenbogen BS, Katzenellenbogen JA. Defining the "S" in SERMs. Science. 2002,295 (5564):2380-2381
[56]Mitlak BH, Cohen FJ. In Search of Optimal Long-Term Female Hormone Replacement: The Potential of Selective Estrogen Receptor Modulators. Hormone Research in Paediatrics.1997,48(4):155-163
[57]Shang Y, Brown M. Molecular Determinants for the Tissue Specificity of SERMs. Science.2002,295 (5564):2465-2468
[58]Purdie, Beardsworth. The selective oestrogen receptor modulation:evolution and clinical applications. British Journal of Clinical Pharmacology.1999,48 (6):785-792
[59]Umemoto A, Lin C-X, Ueyama Y, Komaki K, Santosh Laxmi YR, Shibutani S. Absence of DNA Adduct in the Leukocytes from Breast Cancer Patients Treated with Toremifene. Chemical Research in Toxicology.2006,19 (3):421-425
[60]Shibutani S, Ravindernath A, Terashima I, Suzuki N, Santosh Laxmi YR, Kanno Y, et al. Mechanism of Lower Genotoxicity of Toremifene Compared with Tamoxifen. Cancer Research.2001,61 (10):3925-3931
[61]Gamboa da Costa G, Pereira PC, Churchwell MI, Beland FA, Marques MM. DNA Adduct Formation in the Livers of Female Sprague-Dawley Rats Treated with Toremifene or α-Hydroxytoremifene. Chemical Research in Toxicology.2007,20 (2): 300-310
[62]Erkkola R, Mattila L, Powles T, Heikkinen J, Toivola B, Korhonen P, et al. Bone Mineral Density and Lipid Changes During 5 Years of Follow-up in a Study of Prevention of Breast Cancer with Toremifene in Healthy, High-risk Pre- and Post-menopausal Women. Breast Cancer Research and Treatment.2005,93 (3): 277-287
[63]Harvey HA, Kimura M, Hajba A. Toremifene:An evaluation of its safety profile. The Breast.2006,15 (2):142-157
[64]Kusama M, Kaise H, Nakayama S, Ota D, Misaka T, Aoki T. Crossover trial for lipid abnormality in postmenopausal breast cancer patients during selective estrogen receptor modulators (SERMs) administrations. Breast Cancer Research and Treatment.2004,88 (1):9-16
[65]Kusama M, Miyauchi K, Aoyama H, Sano M, Kimura M, Mitsuyama S, et al. Effects of toremifene (TOR) and tamoxifen (TAM) on serum lipids. Breast Cancer Research and Treatment.2004,88 (1):1-8
[66]Gong C, Song E, Jia W, Qin L, Guo J, Jia H, et al. A Double-blind Randomized Controlled Trial of Toremifen Therapy for Mastalgia. Arch Surg.2006,141 (1):43-47
[67]Bliddal H, Rosetzsky A, Schlichting P, Weidner MS, Andersen LA, Ibfelt HH, et al. A randomized, placebo-controlled, cross-over study of ginger extracts and Ibuprofen in osteoarthritis. Osteoarthritis and Cartilage.2000,8 (1):9-12
[68]Price D, Stein B, Sieber P, Tutrone R, Bailen J, Goluboff E, et al. Toremifene for the Prevention of Prostate Cancer in Men With High Grade Prostatic Intraepithelial Neoplasia:Results of a Double-Blind, Placebo Controlled, Phase ⅡB Clinical Trial. The Journal of Urology.2006,176 (3):965-971
[69]Zhao C, Dahlman-Wright K, Gustafsson JA. Estrogen receptor beta:an overview and update. Nucl Recept Signal.2008,6:e003
[70]Skliris GP, Leygue E, Watson PH, Murphy LC. Estrogen receptor alpha negative breast cancer patients:Estrogen receptor beta as a therapeutic target. The Journal of Steroid Biochemistry and Molecular Biology.2008,109 (1):1-10
[71]Chang EC, Frasor J, Komm B, Katzenellenbogen BS. Impact of Estrogen Receptor beta on Gene Networks Regulated by Estrogen Receptor alpha in Breast Cancer Cells. Endocrinology.2006,147 (10):4831-4842
[72]Murphy LC, Watson PH. Is oestrogen receptor- beta a predictor of endocrine therapy responsiveness in human breast cancer? Endocr Relat Cancer.2006,13 (2):327-334
[73]Rosa FE, Caldeira JRF, Felipes J, Bertonha FB, Quevedo FC, Domingues MAC, et al. Evaluation of estrogen receptor alpha and beta and progesterone receptor expression and correlation with clinicopathologic factors and proliferative marker Ki-67 in breast cancers. Human Pathology.2008,39 (5):720-730
[74]Puddefoot JR, Barker S, Vinson GP. Trilostane in advanced breast cancer. Expert Opinion on Pharmacotherapy.2006,7 (17):2413-2419
[75]Puddefoot JR, Barker S, Glover HR, Malouitre SDM, Vinson GP. Non-competitive steroid inhibition of oestrogen receptor functions. International Journal of Cancer.2002, 101 (1):17-22
[76]Barker S, Malouitre SDM, Glover HR, Puddefoot JR, Vinson GP. Comparison of effects of 4-hydroxy tamoxifen and trilostane on oestrogen-regulated gene expression in MCF-7 cells:up-regulation of oestrogen receptor beta. J Steroid Biochem Mol Biol. 2006,100(4-5):141-151
[77]Yamamoto Y, Shibata J, Yonekura K, Sato K, Hashimoto A, Aoyagi Y, et al. TAS-108, a Novel Oral Steroidal Antiestrogenic Agent, Is a Pure Antagonist on Estrogen Receptor a and a Partial Agonist on Estrogen Receptor β with Low Uterotrophic Effect. Clinical Cancer Research.2005,11 (1):315-322
[78]Buzdar AU. TAS-108:A Novel Steroidal Antiestrogen. Clinical Cancer Research.2005, 11 (2):906-908
[79]Blakely LJ, Buzdar A, Chang H-Y, Frye D, Theriault R, Valero V, et al. A Phase I and Pharmacokinetic Study of TAS-108 in Postmenopausal Female Patients with Locally Advanced, Locally Recurrent Inoperable, or Progressive Metastatic Breast Cancer. Clinical Cancer Research.2004,10 (16):5425-5431
[80]Saeki T, Noguchi S, Aogi K, Inaji H, Tabei T, Ikeda T. Evaluation of the safety and tolerability of oral TAS-108 in postmenopausal patients with metastatic breast cancer. Annals of Oncology.2009,20 (5):868-873
[81]Ariazi EA, Ariazi JL, Cordera F, Jordan VC. Estrogen receptors as therapeutic targets in breast cancer. Current Topics in Medicinal Chemistry 2006,6 (3):181-202
[82]Peng J, Sengupta S, Jordan VC. Potential of selective estrogen receptor modulators as treatments and preventives of breast cancer. Anti-Cancer Agents in Medicinal Chemistry 2009,9 (5):481-499
[83]沈杰.药物ADMET理论预测方法开发和靶向雌激素受体的药物设计研究[博士论文].上海：华东理工大学；2010
[84]Minutolo F, Bertini S, Papi C, Carlson KE, Katzenellenbogen JA, Macchia M. Salicylaldoxime Moiety as a Phenolic "A-Ring" Substitute in Estrogen Receptor Ligands. Journal of Medicinal Chemistry.2001,44 (24):4288-4291
[85]Zhou H-B, Sheng S, Compton DR, Kim Y, Joachimiak A, Sharma S, et al. Structure-Guided Optimization of Estrogen Receptor Binding Affinity and Antagonist Potency of Pyrazolopyrimidines with Basic Side Chains. Journal of Medicinal Chemistry.2006,50 (2):399-403
[86]Fields S, Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989,340 (6230):245-246
[87]Chang B, Elledge SJ, William DG. Gene identification using the yeast two-hybrid system. Methods in Enzymology.1997,283 (273):141-156
[88]Manas ES, Unwalla RJ, Xu ZB, Malamas MS, Miller CP, Harris HA, et al. Structure-Based Design of Estrogen Receptor-β Selective Ligands. Journal of the American Chemical Society.2004,126 (46):15106-15119
[89]Filardo EJ, Quinn JA, Frackelton AR, Jr., Bland KI. Estrogen Action Via the G Protein-Coupled Receptor, GPR30:Stimulation of Adenylyl Cyclase and cAMP-Mediated Attenuation of the Epidermal Growth Factor Receptor-to-MAPK Signaling Axis. Mol Endocrinol.2002,16 (1):70-84
[90]Tang S, Han H, Bajic VB. ERGDB:Estrogen Responsive Genes Database. Nucleic Acids Research.2004,32:D533-D536
[91]Castro-Rivera E, Samudio I, Safe S. Estrogen Regulation of Cyclin D1 Gene Expression in ZR-75 Breast Cancer Cells Involves Multiple Enhancer Elements. Journal of Biological Chemistry.2001,276 (33):30853-30861
[2]Greene GL, Paul G, Michael W, Andrew B, Yvonne H, John S. Sequence and expression of human estrogen receptor complementary DNA. Science.1986,231 (4742): 1150-1154
[3]Green S, Walter P, Kumar V, Krust A, Bornert J-M, Argos P, et al. Human oestrogen receptor cDNA:sequence, expression and homology to v-erb-A. Nature.1986,320 (6058):134-139
[4]Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA. Cloning of a novel receptor expressed in rat prostate and ovary. Proceedings of the National Academy of Sciences of the United States of America.1996,93 (12):5925-5930
[5]Enmark E, Pelto-Huikko M, Grandien K, Lagercrantz S, Lagercrantz J, Fried G, et al. Human estrogen receptor β-Gene structure, chromosomal localization, and expression pattern. J Clin Endocrinol Metab.1997,82 (12):4258-4265
[6]Kong EH, Pike ACW, Hubbard RE. Structure and mechanism of the oestrogen receptor. Biochem Soc Trans.2003,31:56-59
[7]Mosselman S, Polman J, Dijkema R. ER β:Identification and characterization of a novel human estrogen receptor. FEBS Letters.1996,392 (1):49-53
[8]Ogawa S, Inoue S, Watanabe T, Hiroi H, Orimo A, Hosoi T, et al. The complete primary structure of human estrogen receptor beta (hER beta) and its heterodimerization with ER alpha in vivo and in vitro. Biochemical and Biophysical Research Communications. 1998,243(1):122-126
[9]Zhao L, O'Neill K, Diaz Brinton R. Selective estrogen receptor modulators (SERMs) for the brain:Current status and remaining challenges for developing NeuroSERMs. Brain Research Reviews.2005,49 (3):472-493
[10]Kumar V, Green S, Stack G, Berry M, Jin J-R, Chambon P. Functional domains of the human estrogen receptor. Cell.1987,51 (6):941-951
[11]Tora L, White J, Brou C, Tasset D, Webster N, Scheer E, et al. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell.1989, 59 (3):477-487
[12]Berry M, Metzager D, Chambon P. Role of the two activating domains of the oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the anti-oestrogen 4-hydroxytamoxifen. The EMBO Journal.1990,9 (9):2811-2818
[13]Tzukerman M, Esty A, Santiso-Mere D, Danielian P, Parker M, Stein R, et al. Human estrogen receptor transactivational capacity is determined by both cellular and promoter context and mediated by two functionally distinct intramolecular regions. Mol Endocrinol.1994,8 (1):21-30
[14]Fawell SE, White R, Hoare S, Sydenham M, Page M, Parker MG Inhibition of estrogen receptor-DNA binding by the "pure" antiestrogen ICI 164,384 appears to be mediated by impaired receptor dimerization. Proceedings of the National Academy of Sciences of the United States of America.1990,87 (17):6883-6887
[15]Dauvois S, Danielian PS, White R, Parker MG. Antiestrogen ICI 164,384 reduces cellular estrogen receptor content by increasing its turnover. Proceedings of the National Academy of Sciences of the United States of America.1992,89 (9): 4037-4041
[16]Kuiper GGJM, Carlsson B, Grandien K, Enmark E, Haggblad J, Nilsson S, et al. Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology.1997,138 (3):863-870
[17]Pike ACW, Brzozowski AM, Hubbard RE. A structural biologist's view of the oestrogen receptor. The Journal of Steroid Biochemistry and Molecular Biology.2000,74 (5): 261-268
[18]Shiau AK, Barstad D, Loria PM, Cheng L, Kushner PJ, Agard DA, et al. The structural basis of estrogen receptor/coactivator recognition and the antagonism of this interaction by Tamoxifen. Cell.1998,95 (7):927-937
[19]Pike ACW, Brzozowski AM, Hubbard RE, Bonn T, Thorsell A-G, Engstrom O, et al. Structure of the ligand-binding domain of oestrogen receptor beta in the presence of a partial agonist and a full antagonist. EMBO J.1999,18 (17):4608-4618
[20]Pike ACW, Brzozowski AM, Walton J, Hubbard RE, Thorsell A-G, Li Y-L, et al. Structural Insights into the Mode of Action of a Pure Antiestrogen. Structure.2001,9 (2):145-153
[21]McKenna NJ, Lanz RB, O'Malley BW Nuclear receptor coregulators:cellular and molecular biology. Endocr Rev.1999,20 (3):321-344
[22]Klinge CM. Estrogen receptor interaction with co-activators and co-repressors. Steroids. 2000,65 (5):227-251
[23]Tremblay G, Giguere V. Coregualtors of estrogen receptor action. Critical Reviews in Eukaryotic Gene Expression.2002,12 (1):1-22
[24]Heery DM, Kalkhoven E, Hoare S, Parker MG. A signature motif in transcriptional co-activators mediates binding to nuclear receptors. Nature.1997,387 (6634):733-736
[25]Lannigan DA. Estrogen receptor phosphorylation. Steroids.2003,68 (1):1-9
[26]Kato S. Estrogen receptor-mediated cross-talk with growth factor signaling pathways. Breast Cancer.2001,8 (1):3-9
[27]Kousteni S, Bellido T, Plotkin LI, O'Brien CA, Bodenner DL, Han L, et al. Nongenotropic, Sex-Nonspecific Signaling through the Estrogen or Androgen Receptors:Dissociation from Transcriptional Activity. Cell.2001,104 (5):719-730
[28]Nemere I, Pietras RJ, Blackmore PF. Membrane receptors for steroid hormones:Signal transduction and physiological significance. Journal of Cellular Biochemistry.2003,88 (3):438-445
[29]Wang Z, Zhang X, Shen P, Loggie BW, Chang Y, Deuel TF. A variant of estrogen receptor-α, hER-a36:Transduction of estrogen- and antiestrogen-dependent membrane-initiated mitogenic signaling. Proceedings of the National Academy of Sciences.2006,103 (24):9063-9068
[30]Funakoshi T, Yanai A, Shinoda K, Kawano MM, Mizukami Y. G protein-coupled receptor 30 is an estrogen receptor in the plasma membrane. Biochemical and Biophysical Research Communications.2006,346 (3):904-910
[31]Okamoto T, Schlegel A, Scherer PE, Lisanti MP. Caveolins, a Family of Scaffolding Proteins for Organizing reassembled Signaling Complexes at the Plasma Membrane. Journal of Biological Chemistry.1998,273 (10):5419-5422
[32]Guo X, Razandi M, Pedram A, Kassab G, Levin ER. Estrogen Induces Vascular Wall Dilation. Journal of Biological Chemistry.2005,280 (20):19704-19710
[33]Lu Q, Pallas DC, Surks HK, Baur WE, Mendelsohn ME, Karas RH. Striatin assembles a membrane signaling complex necessary for rapid, nongenomic activation of endothelial NO synthase by estrogen receptor. Proceedings of the National Academy of Sciences of the United States of America.2004,101 (49):17126-17131
[34]Castoria G, Migliaccio A, Bilancio A, Di Domenico M, de Falco A, Lombardi M, et al. PI3-kinase in concert with Src promotes the S-phase entry of oestradiol-stimulated MCF-7 cells. EMBO J.2001,20 (21):6050-6059
[35]Fernando RI, Wimalasena J. Estradiol Abrogates Apoptosis in Breast Cancer Cells through Inactivation of BAD:Ras-dependent Nongenomic Pathways Requiring Signaling through ERK and Akt. Mol Biol Cell.2004,15 (7):3266-3284
[36]Weihua Z, Warner M, Gustafsson J-A. Estrogen receptor beta in the prostate. Molecular and Cellular Endocrinology.2002,193 (1-2):1-5
[37]Telleria CM, Zhong L, Deb S, Srivastava RK, Park KS, Sugino N, et al. Differential Expression of the Estrogen Receptors alpha and beta in the Rat Corpus Luteum of Pregnancy:Regulation by Prolactin and Placental Lactogens. Endocrinology.1998,139 (5):2432-2442
[38]Iafrati MD, Karas RH, Aronovitz M, Kim S, Sullivan TR, Lubahn DB, et al. Estrogen inhibits the vascular injury response in estrogen receptor α-deficient mice. Nat Med. 1997,3 (5):545-548
[39]Karas RH, Schulten H, Pare G, Aronovitz MJ, Ohlsson C, Gustafsson J-A, et al. Effects of Estrogen on the Vascular Injury Response in Estrogen Receptor alpha, beta (Double) Knockout Mice. Circ Res.2001,89 (6):534-539
[40]Leav I, Lau K-M, Adams JY, McNeal JE, Taplin M-E, Wang J, et al. Comparative Studies of the Estrogen Receptors beta and alpha and the Androgen Receptor in Normal Human Prostate Glands, Dysplasia, and in Primary and Metastatic Carcinoma. Am J Pathol.2001,159(1):79-92
[41]Peters RK, Pike MC, Chang WWL, Mack TM. Reproductive factors and colon cancers. Br J Cancer.1990,61 (5):741-748
[42]Wenbin L. Age, sex and the risk of grade-specific second primary colorectal cancer: Evidence for the protective effect of female hormone. European Journal of Cancer. 2007,43 (12):1856-1861
[43]Slattery ML, Sweeney C, Murtaugh M, Ma KN, Wolff RK, Potter JD, et al. Associations between ERα, ERβ, and AR Genotypes and Colon and Rectal Cancer. Cancer Epidemiology Biomarkers & Prevention.2005,14 (12):2936-2942
[44]Foley EF, Jazaeri AA, Shupnik MA, Jazaeri O, Rice LW. Selective Loss of Estrogen Receptor β in Malignant Human Colon. Cancer Research.2000,60 (2):245-248
[45]Campbell-Thompson M, Lynch IJ, Bhardwaj B. Expression of Estrogen Receptor (ER) Subtypes and ER β Isoforms in Colon Cancer. Cancer Research.2001,61 (2):632-640
[46]Konstantinopoulos PA, Kominea A, Vandoros G, Sykiotis GP, Andricopoulos P, Varakis I, et al. Oestrogen receptor beta (ER beta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour's dedifferentiation. European Journal of Cancer.2003,39 (9):1251-1258
[47]Castiglione F, Taddei A, Degl Innocenti DR, Buccoliero AM, Bechi P, Garbini F, et al. Expression of Estrogen Receptor β in Colon Cancer Progression. Diagnostic Molecular Pathology.2008,17 (4):231-236
[48]Wada-Hiraike O, Warner M, Gustafsson J-A. New developments in oestrogen signalling in colonic epithelium. Biochem Soc Trans.2006,34 (6):1114-1116
[49]Martineti V, Picariello L, Tognarini I, Sala SC, Gozzini A, Azzari C, et al. ERβ is a potent inhibitor of cell proliferation in the HCT8 human colon cancer cell line through regulation of cell cycle components. Endocr Relat Cancer.2005,12 (2):455-469
[50]Fiorelli G, Picariello L, Martineti V, Tognarini I, Tonelli F, Brandi ML. Estrogen metabolism in human colorectal cancer cells. The Journal of Steroid Biochemistry and Molecular Biology.2002,81 (3):281-289
[51]Di Domenico M, Castoria G, Bilancio A, Migliaccio A, Auricchio F. Estradiol Activation of Human Colon Carcinoma-derived Caco-2 Cell Growth. Cancer Research. 1996,56 (19):4516-4521
[52]Sims NA, Dupont S, Krust A, Clement-Lacroix P, Minet D, Resche-Rigon M, et al. Deletion of estrogen receptors reveals a regulatory role for estrogen receptors-β in bone remodeling in females but not in males. Bone.2002,30 (1):18-25
[53]Lindberg MK, Moverare S, Skrtic S, Gao H, Dahlman-Wright K, Gustafsson J-A, et al. Estrogen Receptor (ER)-β Reduces ER α-Regulated Gene Transcription, Supporting a "Ying Yang" Relationship between ER α and ER β in Mice. Mol Endocrinol.2003,17 (2):203-208
[54]Persson I, Weiderpass E, Bergkvist L, Bergstrom R, Schairer C. Risks of breast and endometrial cancer after estrogen and estrogen-progestin replacement. Cancer Causes and Control.1999,10 (4):253-260
[55]Katzenellenbogen BS, Katzenellenbogen JA. Defining the "S" in SERMs. Science. 2002,295 (5564):2380-2381
[56]Mitlak BH, Cohen FJ. In Search of Optimal Long-Term Female Hormone Replacement: The Potential of Selective Estrogen Receptor Modulators. Hormone Research in Paediatrics.1997,48(4):155-163
[57]Shang Y, Brown M. Molecular Determinants for the Tissue Specificity of SERMs. Science.2002,295 (5564):2465-2468
[58]Purdie, Beardsworth. The selective oestrogen receptor modulation:evolution and clinical applications. British Journal of Clinical Pharmacology.1999,48 (6):785-792
[59]Umemoto A, Lin C-X, Ueyama Y, Komaki K, Santosh Laxmi YR, Shibutani S. Absence of DNA Adduct in the Leukocytes from Breast Cancer Patients Treated with Toremifene. Chemical Research in Toxicology.2006,19 (3):421-425
[60]Shibutani S, Ravindernath A, Terashima I, Suzuki N, Santosh Laxmi YR, Kanno Y, et al. Mechanism of Lower Genotoxicity of Toremifene Compared with Tamoxifen. Cancer Research.2001,61 (10):3925-3931
[61]Gamboa da Costa G, Pereira PC, Churchwell MI, Beland FA, Marques MM. DNA Adduct Formation in the Livers of Female Sprague-Dawley Rats Treated with Toremifene or α-Hydroxytoremifene. Chemical Research in Toxicology.2007,20 (2): 300-310
[62]Erkkola R, Mattila L, Powles T, Heikkinen J, Toivola B, Korhonen P, et al. Bone Mineral Density and Lipid Changes During 5 Years of Follow-up in a Study of Prevention of Breast Cancer with Toremifene in Healthy, High-risk Pre- and Post-menopausal Women. Breast Cancer Research and Treatment.2005,93 (3): 277-287
[63]Harvey HA, Kimura M, Hajba A. Toremifene:An evaluation of its safety profile. The Breast.2006,15 (2):142-157
[64]Kusama M, Kaise H, Nakayama S, Ota D, Misaka T, Aoki T. Crossover trial for lipid abnormality in postmenopausal breast cancer patients during selective estrogen receptor modulators (SERMs) administrations. Breast Cancer Research and Treatment.2004,88 (1):9-16
[65]Kusama M, Miyauchi K, Aoyama H, Sano M, Kimura M, Mitsuyama S, et al. Effects of toremifene (TOR) and tamoxifen (TAM) on serum lipids. Breast Cancer Research and Treatment.2004,88 (1):1-8
[66]Gong C, Song E, Jia W, Qin L, Guo J, Jia H, et al. A Double-blind Randomized Controlled Trial of Toremifen Therapy for Mastalgia. Arch Surg.2006,141 (1):43-47
[67]Bliddal H, Rosetzsky A, Schlichting P, Weidner MS, Andersen LA, Ibfelt HH, et al. A randomized, placebo-controlled, cross-over study of ginger extracts and Ibuprofen in osteoarthritis. Osteoarthritis and Cartilage.2000,8 (1):9-12
[68]Price D, Stein B, Sieber P, Tutrone R, Bailen J, Goluboff E, et al. Toremifene for the Prevention of Prostate Cancer in Men With High Grade Prostatic Intraepithelial Neoplasia:Results of a Double-Blind, Placebo Controlled, Phase ⅡB Clinical Trial. The Journal of Urology.2006,176 (3):965-971
[69]Zhao C, Dahlman-Wright K, Gustafsson JA. Estrogen receptor beta:an overview and update. Nucl Recept Signal.2008,6:e003
[70]Skliris GP, Leygue E, Watson PH, Murphy LC. Estrogen receptor alpha negative breast cancer patients:Estrogen receptor beta as a therapeutic target. The Journal of Steroid Biochemistry and Molecular Biology.2008,109 (1):1-10
[71]Chang EC, Frasor J, Komm B, Katzenellenbogen BS. Impact of Estrogen Receptor beta on Gene Networks Regulated by Estrogen Receptor alpha in Breast Cancer Cells. Endocrinology.2006,147 (10):4831-4842
[72]Murphy LC, Watson PH. Is oestrogen receptor- beta a predictor of endocrine therapy responsiveness in human breast cancer? Endocr Relat Cancer.2006,13 (2):327-334
[73]Rosa FE, Caldeira JRF, Felipes J, Bertonha FB, Quevedo FC, Domingues MAC, et al. Evaluation of estrogen receptor alpha and beta and progesterone receptor expression and correlation with clinicopathologic factors and proliferative marker Ki-67 in breast cancers. Human Pathology.2008,39 (5):720-730
[74]Puddefoot JR, Barker S, Vinson GP. Trilostane in advanced breast cancer. Expert Opinion on Pharmacotherapy.2006,7 (17):2413-2419
[75]Puddefoot JR, Barker S, Glover HR, Malouitre SDM, Vinson GP. Non-competitive steroid inhibition of oestrogen receptor functions. International Journal of Cancer.2002, 101 (1):17-22
[76]Barker S, Malouitre SDM, Glover HR, Puddefoot JR, Vinson GP. Comparison of effects of 4-hydroxy tamoxifen and trilostane on oestrogen-regulated gene expression in MCF-7 cells:up-regulation of oestrogen receptor beta. J Steroid Biochem Mol Biol. 2006,100(4-5):141-151
[77]Yamamoto Y, Shibata J, Yonekura K, Sato K, Hashimoto A, Aoyagi Y, et al. TAS-108, a Novel Oral Steroidal Antiestrogenic Agent, Is a Pure Antagonist on Estrogen Receptor a and a Partial Agonist on Estrogen Receptor β with Low Uterotrophic Effect. Clinical Cancer Research.2005,11 (1):315-322
[78]Buzdar AU. TAS-108:A Novel Steroidal Antiestrogen. Clinical Cancer Research.2005, 11 (2):906-908
[79]Blakely LJ, Buzdar A, Chang H-Y, Frye D, Theriault R, Valero V, et al. A Phase I and Pharmacokinetic Study of TAS-108 in Postmenopausal Female Patients with Locally Advanced, Locally Recurrent Inoperable, or Progressive Metastatic Breast Cancer. Clinical Cancer Research.2004,10 (16):5425-5431
[80]Saeki T, Noguchi S, Aogi K, Inaji H, Tabei T, Ikeda T. Evaluation of the safety and tolerability of oral TAS-108 in postmenopausal patients with metastatic breast cancer. Annals of Oncology.2009,20 (5):868-873
[81]Ariazi EA, Ariazi JL, Cordera F, Jordan VC. Estrogen receptors as therapeutic targets in breast cancer. Current Topics in Medicinal Chemistry 2006,6 (3):181-202
[82]Peng J, Sengupta S, Jordan VC. Potential of selective estrogen receptor modulators as treatments and preventives of breast cancer. Anti-Cancer Agents in Medicinal Chemistry 2009,9 (5):481-499
[83]沈杰.药物ADMET理论预测方法开发和靶向雌激素受体的药物设计研究[博士论文].上海：华东理工大学；2010
[84]Minutolo F, Bertini S, Papi C, Carlson KE, Katzenellenbogen JA, Macchia M. Salicylaldoxime Moiety as a Phenolic "A-Ring" Substitute in Estrogen Receptor Ligands. Journal of Medicinal Chemistry.2001,44 (24):4288-4291
[85]Zhou H-B, Sheng S, Compton DR, Kim Y, Joachimiak A, Sharma S, et al. Structure-Guided Optimization of Estrogen Receptor Binding Affinity and Antagonist Potency of Pyrazolopyrimidines with Basic Side Chains. Journal of Medicinal Chemistry.2006,50 (2):399-403
[86]Fields S, Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989,340 (6230):245-246
[87]Chang B, Elledge SJ, William DG. Gene identification using the yeast two-hybrid system. Methods in Enzymology.1997,283 (273):141-156
[88]Manas ES, Unwalla RJ, Xu ZB, Malamas MS, Miller CP, Harris HA, et al. Structure-Based Design of Estrogen Receptor-β Selective Ligands. Journal of the American Chemical Society.2004,126 (46):15106-15119
[89]Filardo EJ, Quinn JA, Frackelton AR, Jr., Bland KI. Estrogen Action Via the G Protein-Coupled Receptor, GPR30:Stimulation of Adenylyl Cyclase and cAMP-Mediated Attenuation of the Epidermal Growth Factor Receptor-to-MAPK Signaling Axis. Mol Endocrinol.2002,16 (1):70-84
[90]Tang S, Han H, Bajic VB. ERGDB:Estrogen Responsive Genes Database. Nucleic Acids Research.2004,32:D533-D536
[91]Castro-Rivera E, Samudio I, Safe S. Estrogen Regulation of Cyclin D1 Gene Expression in ZR-75 Breast Cancer Cells Involves Multiple Enhancer Elements. Journal of Biological Chemistry.2001,276 (33):30853-30861