Current Protocols
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2024
·Vol. 4
·期 4
·e1033
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IF 0
Ruifeng Wang,Shuhua Ma,Nengwei Xu,Yumiao Gan,Pengya Li,Jingying Zhang,Zhixiang Zhang,Qingyang Gu,Jian Xiang
Prostate cancer is a leading diagnosis and major cause of cancer-related deaths in men worldwide. As a typical hormone-responsive disease, prostate cancer is commonly managed with androgen deprivation therapy (ADT) to curb its progression and potential metastasis. Unfortunately, progression to castration-resistant prostate cancer (CRPC), a notably more aggressive phase of the disease, occurs within a timeframe of 2-3 years following ADT. Enzalutamide, a recognized androgen receptor (AR) antagonist, has been employed as a standard of care for men with metastatic castration-resistant prostate cancer (mCRPC) since it was first approved in 2012, due to its ability to prolong survival. However, scientific evidence suggests that sustained treatment with AR antagonists may induce acquired AR mutations or splice variants, such as AR F877L, T878A, and H875Y, leading to drug resistance and thereby diminishing the therapeutic efficacy of these agents. Thus, the establishment of prostate cancer models incorporating these particular mutations is essential for developing new therapeutic strategies to overcome such resistance and evaluate the efficacy of next-generation AR-targeting drugs. We have developed a CRISPR (clustered regularly interspaced short palindromic repeats)-based knock-in technology to introduce an additional F877L mutation in AR into the human prostate cell line LNCaP. This article provides comprehensive descriptions of the methodologies for cellular gene editing and establishment of an in vivo model. Using these methods, we successfully identified an enzalutamide-resistant phenotype in both in vitro and in vivo models. We also assessed the efficacy of target protein degraders (TPDs), such as ARV-110 and ARV-667, in both models, and the corresponding validation data are also included here. © 2024 Wiley Periodicals LLC. Basic Protocol 1 : Generation of AR F877L-mutated LNCaP cell line using CRISPR technology Basic Protocol 2 : Validation of drug resistance in AR F877L-mutated LNCaP cell line using the 2D CTG assay Support Protocol : Testing of sgRNA efficiency in HEK 293 cells Basic Protocol 3 : Validation of drug resistance in AR F877L-mutated LNCaP cell line in vivo
研究领域:
肿瘤学
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分子生物学
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基因编辑
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药理学
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DOI
Drug Design Development and Therapy
·
2026
·
IF 6.1
Shengnan Wang,Shijia Lu,Guofei Li,Yingjie Qi
Purpose Hepatocellular carcinoma (HCC) is a highly lethal malignancy with poor prognosis and limited clinical treatments. CD8⁺ T cell-based immunotherapy offers promising therapeutic potential against HCC. We fabricated MMP-9/pH dual-sensitive liposomes co-encapsulating enzalutamide, GB-1107 and BMS-1166 for HCC treatment. Reliable detection approaches are lacking to assess in vivo liposomal behavior. This study established an HPLC-MS/MS method for simultaneous determination of the three drugs in rat plasma, and applied it to characterize their pharmacokinetic profiles.Methods The pH/MMP-9 dual-sensitive liposomes co-loaded with BMS-1166, GB-1107 and enzalutamide were prepared via the thin-film hydration method, and the resultant formulations exhibited satisfactory pharmaceutical properties meeting the predefined quality criteria. For chromatographic separation, a poroshell 120 EC-C18 column (50 × 4.6 mm, 2.7 μm) was adopted, combined with linear gradient elution of 0.1% formic acid in water and acetonitrile. Full validation of the established method was performed strictly following the guidelines for bioanalytical method validation as stated in the Chinese Pharmacopoeia.Results BMS-1166, GB-1107, and enzalutamide exhibited good linearity over the concentration range of 1.0–1000.0 ng/mL. The accuracy and precision of BMS-1166, GB-1107, and enzalutamide all fall below 11.1%. In addition, the recoveries and matrix effect for these analytes ranged from 88.6%-99.7%. Pharmacokinetic results demonstrated that the prepared liposomes possessed favorable in vivo long-circulating properties, which prolonged the half-lives of BMS-1166 and GB-1107 by 25-fold and 13-fold, respectively, and enhanced their bioavailability by 35-fold and 30-fold, respectively. In contrast, no statistically significant effect was observed on enzalutamide.Conclusion The established method was validated and met the predefined quality criteria, thereby being suitable for investigating the in vivo pharmacokinetic profiles of BMS-1166, GB-1107, and enzalutamide.
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产品关联
DOI
PubMed
高影响因子
CHEMICO-BIOLOGICAL INTERACTIONS
·
2025
·
IF 5.4
Tingting Zheng,Hang Yin,Hongna Shan,Lihuan Sun,Lili Jiang,Yong Liu
Enzalutamide (formerly called MDV3100), a second-generation androgen receptor inhibitor primarily used for treating castration-resistant prostate cancer (CRPC), has been shown to regulating UDP-glucuronosyltransferases (UGTs) expression, while its effects on the activity of UGT isoforms remain unclear. This study aimed to systemically investigate the effects of enzalutamide and its phase I metabolite on the UGT activities, and to assess the risk of drug-drug interactions (DDIs) resulting from UGT inhibition. Using high-performance liquid chromatography (HPLC) and enzyme kinetic studies, we evaluated the inhibitory effects of enzalutamide and its main phase I metabolite, N-desmethyl enzalutamide on 12 UGT isoforms and inhibition patterns. Our findings demonstrated that both enzalutamide and its metabolite exhibited potent inhibition against UGT1A1, with evidence of synergistic effects. Enzyme kinetic analysis revealed that both enzalutamide and its metabolite acted as competitive inhibitors of UGT1A1. In vitro-in vivo extrapolation (IVIVE) prediction indicated that concomitant use of enzalutamide with UGT1A1 substrates could induce clinically significant DDIs. Therefore, special caution should be warranted when administering enzalutamide alongside other drugs metabolized by UGT to mitigate potential DDIs.
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DOI
PubMed
高影响因子
Scientific Reports
·
2018
·Vol. 8
·期 1
·1-11
·
IF 4.122
Jiang Zhihui,Zhou Bo,Li Xinping,Kirby Gordon M.,Zhang Xiaoying
Male infertility is a major health issue with an estimated prevalence of 4.2% of male infertility worldwide. Our early work demonstrated that Cistanche extracts protect against sperm damage in mice and that echinacoside (ECH) is one of the major active components. Here we report an essential role for ECH, a natural product that reverses or protects against oligoasthenospermia in rats. ECH was assayed by HPLC, the quantity and quality of sperm was evaluated and hormone levels were determined by radioimmunosorbent assay. ECH reduced levels of androgen receptor (AR) and key steroidogenic-related genes as determined by Western blot and qPCR analysis. The interaction between ECH and AR were evaluated by indirect ELISA and molecular docking. The results show that ECH combined with hypothalamic AR in the pocket of Met-894 and Val-713 to inhibit transfer of AR from the cytoplasm to nuclei in the hypothalamus. While negative feedback of sex hormone regulation was inhibited, positive feedback was stimulated to increase the secretion of luteinizing hormone and testosterone subsequently enhancing the quantity of sperm. Taken together, these data demonstrate that ECH blocks AR activity in the hypothalamus to increase the quantity of sperm and protect against oligoasthenospermia in rats.
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产品关联
DOI
PubMed
高影响因子
Materials Today Bio
·
2025
·
IF 10.2
Shijia Lu,Chenxiao Zhang,Huiru Wu,Junjie Wang,Jinglong Wang,Limei Zhao,Tianyang Ren,Guofei Li
Liver cancer is one of the malignant tumors with high morbidity and mortality worldwide and lacks effective clinical treatments. CD8 + T cell-based immunotherapy is the most promising option for the treatment of liver cancer, but its application is severely limited by low infiltration and functional exhaustion of CD8 + T cells in liver cancer tissues. To overcome this problem, in this study, pH/matrix metalloproteinase-9 (MMP-9) dual-responsive liposomes were constructed and co-loaded with a galectin-3 (Gal-3) inhibitor, GB1107, a programmed death receptor 1 (PD-1) inhibitor, BMS1166, and an androgen receptor (AR) antagonist, enzalutamide, for immunotherapy studies in liver cancer. Specifically, the co-loaded liposomes (GBE@LP) were produced with a one-two punch targeting antitumor immunity. First, GBE@LP exhibited prolonged circulation in the blood and accumulated in liver cancer tissues through the enhanced permeability and retention effect due to the presence of surface polyethylene glycol. After entering liver cancer tissues, GBE@LP responded to the low pH and overexpression of MMP-9 in the microenvironment to promote the release of GB1107, BMS1166, and enzalutamide, thus, realizing the intelligent release of drugs targeting liver cancer. Upon reaching liver tumor tissues, GB1107 can inhibit overexpressed Gal-3 to transform liver cancer from a “cold” to a “hot” tumor and enable deep penetration by CD8 + T cells. In addition, BMS1166 and enzalutamide inhibit PD-1 and AR in CD8 + T cells, reversing CD8 + T cell exhaustion and enhancing their killing activity. In vivo and in vitro experiments confirmed the significant antitumor effects of GBE@LP in liver cancer. These results indicate that our strategy is expected to reverse the “cold” tumor and immune exhaustion state of liver cancer, which not only improves therapeutic efficacy, but also provides a new paradigm for the exploration of liver cancer immunotherapy.
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PubMed
高影响因子
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