Enzalutamide, an AR signaling inhibitor, is effective for bicalutamide-resistant tumors and has been used in metastatic castration-resistant prostate cancer. Enzalutamide (MDV3100)是一种雄激素受体(AR)拮抗剂，IC50为36 nM。
Enzalutamide is an androgen receptor inhibitor and antagonizes induction of prostate-specific antigen (PSA) and transmembrane serine protease 2 (TMPRSS2) in combination with the synthetic androgen R1881 in parental LNCaP cells. Enzalutamide prevents nuclear translocation and co-activator recruitment of the ligand-receptor complex.
The inhibition of cell proliferation
The binding affinity
2% DMSO+30% PEG300
Effect of Compounds on AR by Measuring Secreted Levels of Prostate Specific Antigen (PSA)
PSA levels are indicators of AR activities in prostate cancer.LNCaP-AR cells were maintained in Iscove's medium containing 10% FBS. Two days prior to drug treatment, the cells were grown in Iscove's medium containing 10% CS-FBS to deprive of androgens. The cells were split and grown in Iscove's medium containing 10% CS-FBS with appropriate concentrations of R1881 and the test compounds. After four days incubation, secreted PSA levels were assayed using PSA ELISA kits.
LNCaP cell proliferation assay
LNCaP cells were cultured in RPMI-1640 supplemented with 10% Fetal Bovine Serum (FBS) at 37oC in a 5% CO2humidified incubator. All experiments were performed in triplicate or more. Cells were trypsinized and diluted to 100,000 cells/mL with RPMI-1640 supplemented with 10% FBS. This cell suspension was seeded in 96-well plates at a volume of 100μL (10,000 cells/well) and incubated at 24 h. After removal of 100μL of medium from each well, 200μL of the drug solution, which was supplemented with serial dilutions of the test compounds or DMSO, was added. Then the plates were incubated at 37oC under 5% CO2for 3 d. A 20μL aliquot of MTT (5 mg/mL) was added to each well of microcultures, and the cells were incubated for 4 h. carefully remove the supernatant, and then add 150μL of DMSO so as to melt the crystal. The absorbance at 492 nm was measured. This parameter relates to the number of living cells in the culture.
Drugs, reagents and animals
Dihydrotestosterone (DHT), PG-21anti-AR antibody, Enzalutamide were prepared. Female NOD scid gamma mice, aged 6–8 weeks on arrival, were purchased and housed under standard conditions. All experiments were conducted under an Institutional Animal Care and Use Committee (IACUC)-approved protocol.
The human ovarian adenocarcinoma cell line OVCAR-3 was obtained and cultured in RPMI-1640 medium with 10% fetal bovine serum and insulin, and kept at 37°C in a humidified 5% CO2atmosphere.
In vivo tumor growth assessment
Mice were implanted subcutaneously with approximately 10 × 106OVCAR-3 cells along with Matrigeldivided into four groups (n = 6 per group) and treated with either enzalutamide (30 mg/kg); DHT (12.5 mg/60 days); enzalutamide + DHT, or vehicle control for 7 weeks. The same experiment was repeated using a higher dose of enzalutamide (50 mg/kg) and a larger cohort size (n = 15 per group). Enzalutamide was administered daily per os (PO) once the mean tumor size reached 100 mm3in each cohort. DHT was administered as pellets set to release 12.5 mg DHT over 60 days (approximately 0.21 mg/day), and was implanted 5 days prior to the start of the experiment according to the manufacturer’s instructions. Tumors were measured twice weekly until day 56, and tumor volumes were calculated using the following formula: length × width2× 0.52.
动物 A (mg/kg) = 动物 B (mg/kg)×动物 B的Km系数/动物 A的Km系数
例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。
 Zuo M, Xu X, Xie Z, et al. Design and synthesis of indoline thiohydantoin derivatives based on enzalutamide as antiproliferative agents against prostate cancer. Eur J Med Chem. 2017;125:1002-1022.
 Tran C, Ouk S, Clegg NJ, et al. Development of a second-generation antiandrogen for treatment of advanced prostate cancer. Science. 2009;324(5928):787-790.
|NCT Number||Conditions||Interventions||Sponsor/Collaborators||Phases||Start Date||Last Updated|
|NCT01902251||Prostate Cancer|Pharmacokinetics of Enzalutamide||Drug: Enzalutamide tablet|Drug: Enzalutamide capsule||Astellas Pharma Europe B.V.|Medivation, Inc.|Astellas Pharma Inc||Phase 1||2012-11-01||2014-09-08|
|NCT01927627||Adenocarcinoma of the Prostate||Drug: enzalutamide||Case Comprehensive Cancer Center||Phase 2||2013-11-01||2016-09-15|
|NCT02124668||Castration-Resistant Prostate Cancer|Prostate Cancer||Drug: Enzalutamide||Astellas Pharma Europe B.V.|Medivation, Inc.|Astellas Pharma Inc||Phase 2||2014-09-01||2016-12-22|
|NCT02953860||Breast Cancer||Drug: Fulvestrant with Enzalutamide||University of Colorado, Denver|United States Department of Defense||Phase 2||2017-01-01||2017-01-26|
|NCT02500901||Metastatic Prostate Cancer||Drug: Enzalutamide|Drug: Niraparib||Paul Mathew, MD|Hoosier Cancer Research Network|Tesaro, Inc.||Phase 1||2016-03-01||2017-02-06|
|NCT02138162||Severe Hepatic Impairment|Normal Hepatic Function||Drug: enzalutamide||Astellas Pharma Europe B.V.|Medivation, Inc.|Astellas Pharma Inc||Phase 1||2013-08-01||2014-05-12|
|NCT01911741||Healthy Subjects|Castration Resistant Prostate Cancer (CRPC)|Relative Bioavailability|MDV3100||Drug: MDV3100||Astellas Pharma Europe B.V.|Medivation, Inc.|Astellas Pharma Inc||Phase 1||2012-11-01||2013-07-26|
|NCT02605863||Bladder Cancer||Drug: Enzalutamide||University of Rochester|Astellas Pharma Inc||Phase 2||2016-01-01||2017-01-12|
|NCT02138799||Drug-Drug Interaction (DDI)|Healthy Subjects|Pharmacokinetics of Enzalutamide||Drug: enzalutamide|Drug: rifampin||Astellas Pharma Europe B.V.|Medivation, Inc.|Astellas Pharma Inc||Phase 1||2013-07-01||2014-10-14|
|NCT02028988||Intermediate Risk Prostate Cancer||Drug: Enzalutamide|Radiation: External Beam Radiation||Dana-Farber Cancer Institute|Medivation, Inc.||Phase 2||2014-01-01||2017-02-13|
|NCT02684227||Endometrial Cancer||Drug: Enzalutamide|Drug: Carboplatin|Drug: Paclitaxel||M.D. Anderson Cancer Center|Astellas Pharma Inc|Medivation, Inc.|National Comprehensive Cancer Network||Phase 2||2016-08-01||2016-12-01|
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|NCT02288936||Hormone-refractory Prostate Cancer||Drug: Enzalutamide||Spanish Oncology Genito-Urinary Group||Phase 2||2015-02-01||2017-02-28|
|NCT02207504||Castration-resistant Prostate Cancer||Drug: Crizotinib|Drug: Enzalutamide||Dana-Farber Cancer Institute|Astellas Pharma Inc|Pfizer||Phase 1||2014-08-01||2017-01-29|
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