| 名称 | V-9302 |
| 描述 | V-9302 is a competitive antagonist of transmembrane glutamine flux. V-9302 selectively targets the amino acid transporter ASCT2 (SLC1A5) without affecting ASCT1. V-9302 inhibits ASCT2-mediated glutamine uptake in HEK-293 cells (IC50=9.6 μM). V-9302 can be used for research on tumor and amino acid transport |
| 细胞实验 | Live-cell amino acid uptake assays using HEK293 cells were carried out in 96-well plates. 96-well plates were coated with poly-D-lysine prior to the assay. Cells were plated at a density of 35,000 cells per well 24 h prior to carrying out the assay. Each set of conditions was replicated at least three times, technically and biologically. Cells were washed three times with 100 μL of assay buffer (containing 137 mM NaCl, 5.1 mM KCl, 0.77 mM KH2PO4, 0.71 mM MgSO4.7H2O, 1.1 mM CaCl2, 10 mM D-glucose, and 10 mM HEPES) to remove cell media. 3H-amino acid (500 nM) in the same buffer was added concomitantly with V-9302 and allowed to incubate for 15 min at 37 oC. For ASCT2-mediated 3H-glutamine uptake assays, 5 mM of the system-L inhibitor 2-amino-2-norbornanecarboxylic acid (BCH) was added and the assay buffer was adjusted to pH 6.0. For selectivity studies, no BCH was added and the assay was conducted at pH 7.4. Following the incubation period, the 3H-glutamine/inhibitor was removed and the cells were washed three times with assay buffer. The cells were then lysed by the addition of 50 μL of 1 M NaOH. For reading, 150 μL of scintillation fluid was added and the plates were counted on a scintillation counter. IC50 was calculated using GraphPad Prism. |
| 动物实验 | Animal handling methods for PET imaging studies were conducted as reported. Prior to imaging, animals were allowed to acclimate to facility environment for at least 1 h in a warmed chamber at 31.5 °C. Animals were administered 10.4–11.8 MBq 4-[18F]fluoroglutamine via intravenous injection and imaged using a scanner. During imaging, animals were maintained under 2% isoflurane anesthesia in oxygen at 2 L/min and kept warm for the duration of the PET scan. PET images in xenograft-bearing mice were acquired as 60-minute dynamic data sets. Imaging was initiated three hours post-treatment following vehicle or V-9302 (75 mg/kg) administration. PET data were reconstructed using a three-dimensional (3D) ordered subset expectation maximization/maximum a posteriori (OSEM3D/MAP) algorithm. The resulting three-dimensional reconstructions had an x-y voxel size of 0.474 mm and inter-slice distance of 0.796 mm. ASIPro software was used to manually draw 3D regions of interest (ROIs) surrounding the entire tumor volume. 4-[18F]fluoroglutamine uptake was quantified as the percentage of the injected dose per gram of tissue (%ID/g). Significance was calculated using a t-test in Graphpad Prism. Error is reported as standard deviation (SD). |
| 体外活性 | 方法：人胰腺癌细胞 BxPC-3， HPAC 加入 V-9302 单药 0–20 μM （组合成剂量矩阵），处理 48 h，MTT 法检测细胞活力。
结果：V-9302 增加 caspase 7 剪切、PARP 剪切和 PUMA 表达，提示诱导凋亡。[1]
方法：采用 MTT 法与克隆形成实验，检测 0.1、1、5、10 μM V-9302 单独处理对 BxPC-3、HPAC 胰腺癌细胞活力与克隆形成的影响。
结果：V-9302 可抑制胰腺癌细胞增殖。[2]
方法：正常人肺成纤维细胞（NHLF）、特发性肺纤维化（IPF）成纤维细胞、小鼠 AKR-2B 成纤维细胞，加入 10 μM V-9302 预处理 1h，联合 5 ng/ml TGF-β处理 48h，qPCR、Western blot 检测 SLC1A5 及纤维化标志物。
结果：V-9302 显著抑制谷氨酰胺摄取，阻断 TGF-β 诱导的 Col1、FN、CTGF、ACTA2 等促纤维化蛋白表达。[3] |
| 体内活性 | 方法：C57BL/6 小鼠气管注射博莱霉素（3.5 U/kg）造模，第 11 天起，腹腔注射 V-9302 37.5 mg/kg，每天一次，连续 13 天。
结果：V-9302 显著改善肺顺应性、提升血氧饱和度，降低肺重与胶原沉积，减轻肺组织炎症浸润与纤维化程度，下调 Col1a1、Fn、CTGF 等促纤维化基因，抑制肺组织 mTOR、HIF1/2α、c-Myc 激活，无明显肝肾毒性与体重下降。[3] |
| 存储条件 | Store at low temperature,Keep away from moisture
Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | H2O : Insoluble
DMSO : 11 mg/mL (20.42 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 10 mg/mL (18.56 mM), Solution.
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| 关键字 | V-9302 | V9302 | V 9302 | uptake | transporter | transmembrane glutamine flux | stress | proliferation | oxidative | nude | mice | Inhibitor | inhibit | HEK-293 | glutamine | flux | athymic |
| 相关产品 | GPNA hydrochloride | SN05 | D-Isoleucine | BMS-466442 | L-4FPG | V-9302 hydrochloride | Idactamab | ASCT2-IN-1 | SN40 | (R)-KMH-233 | Agnuside |
| 相关库 | 抑制剂库 | 经典已知活性库 | 已知活性化合物库 | 谷氨酰胺代谢化合物库 | NO PAINS 化合物库 | 抗COVID-19化合物库 |
