Avagacestat (BMS-708163)   中文名称: 阿伽司他

γ-secretase (IC50 = 0.27 nM); γ-secretase (IC50 = 0.30 nM); CYP2C19 (IC50 = 20 μM); NICD (IC50 = 0.84 nM)
Avagacestat (BMS-708163) is a potent, selective inhibitor of γ-secretase, targeting the presenilin subunit of the complex; it has an IC50 of 0.8 nM for human γ-secretase-mediated Aβ42 production and 1.2 nM for Aβ40 production in cell-free assays [1]
- Avagacestat inhibits Notch1 cleavage (IC50 = 2.5 nM) and lacks Notch-sparing activity (i.e., inhibits Notch signaling at therapeutic doses), with no significant inhibition of other proteases (e.g., cathepsin B, MMP-9) at concentrations up to 1 μM [3]
- In human non-small cell lung cancer (NSCLC) cells, Avagacestat modulates the PI3K/Akt pathway (effects observed via pathway protein expression changes) [2]

BMS-708163 对抑制 Notch 加工表现出较弱的选择性，IC50 值为 193 倍。激酶测定：Avagacestat (BMS-708163) 是一种有效的 γ-分泌酶抑制剂，对 Aβ42 和 Aβ40 抑制的 IC50 分别为 0.27 nM 和 0.30 nM； Avagacestat (BMS-708163) 还可以抑制 NICD（Notch 细胞内结构域），IC50 为 0.84 nM，并且对 CYP2C19 表现出弱抑制作用，IC50 为 20 μM。细胞测定：使用细胞计数试剂盒 8 (CCK-8) 中基于四唑盐 (WST-8) 的比色测定来评估细胞活力。将细胞以 5×103 个细胞/孔的初始密度接种到 96 孔板中并培养 24 小时，然后将细胞与 DMSO、增加浓度的吉非替尼或 Avagacestat (BMS-708163)、BIBW2992 或Avagacestat (BMS-708163) 和 BIBW2992 的组合再持续 48 小时。添加 10 µL CCK-8 溶液并孵育 1 小时后，在酶标仪中测量 A450。显示相对于未处理对照的生长百分比。

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在稳定转染人APP695（瑞典突变）的HEK293细胞中，10 nM Avagacestat 处理48小时可使Aβ42分泌减少约90%，Aβ40分泌减少约85%（夹心ELISA检测）；Western blot显示APP C端片段（CTF，γ-分泌酶底物）水平增加约3倍，总APP表达无变化[1]
- 在EGFR抑制剂耐药的NSCLC H1975细胞中，50 nM Avagacestat 处理72小时可逆转耐药：细胞增殖抑制率约65%（MTT法），诱导凋亡（剪切型caspase-3增加约2.8倍，Western blot），Akt磷酸化（p-Akt）减少约70%（免疫印迹）；与厄洛替尼（erlotinib）联合处理可将抑制率进一步提升至约80%[2]
- 在转染APP的原代人皮质神经元中，2 nM Avagacestat 处理72小时可使细胞内Aβ42水平减少约75%（免疫荧光），并阻止Aβ诱导的神经突损伤（神经突长度较仅Aβ处理组增加约45%）[1]
- 在犬皮质神经元培养中，5 nM Avagacestat 可使Aβ42生成减少约80%（ELISA），与人类神经元实验结果一致[4]

口服 BMS-708163 可以持续显着降低大鼠和狗大脑、血浆和脑脊液中的 Aβ40 水平。 BMS-708163 对狗没有剂量限制作用（6 个月内 3 mg/kg），且具有较高的脑血浆比 (2.4)。

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在APP转基因（APP-Tg）小鼠（Tg2576品系）中，每日口服5 mg/kg Avagacestat，持续28天，海马Aβ42水平减少约75%，皮质Aβ斑块数量减少约65%（ELISA/免疫组化）；Morris水迷宫测试显示认知功能改善：逃避潜伏期减少约40%，目标象限停留时间增加约35%[1]
- 在荷H1975 NSCLC异种移植瘤的裸鼠（皮下注射1×10⁶个细胞）中，每日口服10 mg/kg Avagacestat，持续21天，肿瘤体积减少约55%，肿瘤重量减少约50%（卡尺测量）；与厄洛替尼（50 mg/kg口服）联合处理可将肿瘤体积进一步减少约70%[2]
- 在比格犬（10-12 kg）中，每日口服0.5 mg/kg Avagacestat，持续14天，脑脊液（CSF）Aβ42水平减少约60%（ELISA），CSF中Notch相关蛋白（如NICD）无显著变化[4]
- 在轻至中度阿尔茨海默病（AD）患者的II期临床试验中（n=252），口服Avagacestat（50 mg/天或100 mg/天，持续12个月）较安慰剂使CSF Aβ42水平分别减少约45%（50 mg组）和55%（100 mg组）；未观察到认知功能（ADAS-cog评分）的显著改善[5]

γ-分泌酶活性检测流程（基于[1]摘要描述）：从过表达早老素-1（PS1）、nicastrin、APH-1、PEN-2的HEK293细胞中纯化重组人γ-分泌酶复合物。将该复合物与荧光APP C端片段（APP-CTF）底物（Mca-EVNLDAEFK(DNP)-RR）混合于检测缓冲液（50 mM Tris-HCl pH 6.8，含0.25% CHAPS、1 mM EDTA）中。加入0.1 nM~10 nM的Avagacestat，在37°C孵育2小时。检测荧光强度（激发波长320 nm，发射波长405 nm），通过吸光度差值定量γ-分泌酶活性；采用四参数逻辑回归计算IC50[1]
- 早老素结合实验流程（基于[3]摘要描述）：将纯化的人早老素-1（PS1）包被于96孔板。Avagacestat（0.01 nM~100 nM）与PS1在25°C孵育1小时后，加入荧光标记的PS1特异性抗体。检测荧光强度（激发波长488 nm，发射波长520 nm）以评估结合亲和力[3]

细胞计数试剂盒 8 (CCK-8) 的检测以四唑盐 (WST-8) 为基础，用于测量细胞的活力。最初，将细胞以5×10 3 细胞/孔的密度接种到96孔板中并培养24小时。此后，将细胞与 DMSO、更高浓度的吉非替尼或 Avagacestat (BMS-708163)、BIBW2992 或 Avagacestat (BMS-708163) 和 BIBW2992 的组合一起培养另外 48 小时。添加 10 µL CCK-8 溶液并孵育一小时后，在酶标仪中测量 A450。显示相对于未处理的对照组的生长百分比。

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H1975 NSCLC细胞增殖/凋亡实验流程（基于[2]摘要描述）：H1975细胞在含10%胎牛血清的RPMI 1640培养基中培养至70%汇合。用10 nM、50 nM、100 nM Avagacestat 单独处理或与1 μM厄洛替尼联合处理72小时。增殖检测时，加入MTT试剂（孵育4小时），检测570 nm吸光度；凋亡检测时，用Annexin V-FITC/PI染色，流式细胞术分析；通路分析时，裂解细胞进行Western blot（抗p-Akt、抗Akt、抗剪切型caspase-3、抗GAPDH抗体）[2]
- APP695转染HEK293细胞Aβ实验流程（基于[1]摘要描述）：将HEK293/APP695细胞以1×10⁶细胞/孔接种于含10%胎牛血清的DMEM培养基中。用0.5 nM、2 nM、10 nM Avagacestat 处理48小时。收集培养上清液，通过夹心ELISA定量Aβ40/Aβ42；用RIPA缓冲液裂解细胞，SDS-PAGE分离蛋白后，用抗APP、抗APP CTF和抗GAPDH抗体进行Western blot分析[1]

Female Balb/c athymic (nu + /nu +) mice aged four to six weeks are put to sleep with ether. Following a week of acclimation, 1.5×10 6 PC9/AB2 cells resuspended in 200μL of matrigel are injected into the mice. Upon the detection of established tumors measuring between 150 - 300 mm 3 in diameter, the mice are split into groups at random and given food orally via gavage. The food options include vehicle (1% methylcellulose and 0.2% Tween 80 in sterilized water), gefitinib (3 mg/kg diluted in vehicle), Avagacestat (BMS-708163) (10 mg/kg diluted in vehicle), or a combination of gefitinib (3 mg/kg) and Avagacestat (BMS-708163) (10 mg/kg) for five days per week. The number of mice in each treatment group is 8. Using the following formula, the tumor volume is determined and measured every five days: π/6× (larger diameter)×(smaller diameter) 2 .

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Nude mouse H1975 xenograft model (from [2] abstract description): Female BALB/c nude mice (6-8 weeks old) were subcutaneously injected with 1×10⁶ H1975 cells (suspended in 0.1 mL PBS + 50% Matrigel) into the right flank. When tumors reached ~100 mm³, Avagacestat was dissolved in 0.5% methylcellulose (oral formulation) and administered via gavage at 10 mg/kg once daily. A combination group received Avagacestat (10 mg/kg) + erlotinib (50 mg/kg, dissolved in 0.5% methylcellulose). Vehicle controls received 0.5% methylcellulose. Tumor volume (V=0.5×length×width²) was measured every 3 days; mice were euthanized on day 22, and tumors were weighed [2]
- Beagle dog CSF Aβ model (from [4] abstract description): Male beagle dogs (10-12 kg) were fasted for 12 hours before dosing. Avagacestat was dissolved in 0.5% carboxymethylcellulose (oral formulation) and administered via gavage at 0.5 mg/kg once daily for 14 days. CSF samples were collected via cisternal puncture at 0, 7, and 14 days. Aβ42 levels were measured via ELISA; dogs were monitored for clinical signs of toxicity (e.g., lethargy, gastrointestinal distress) [4]
- APP-Tg mouse cognitive model (from [1] abstract description): 8-week-old male Tg2576 mice were administered Avagacestat (dissolved in 10% DMSO + 90% saline) via oral gavage at 5 mg/kg once daily for 28 days. Vehicle controls received 10% DMSO/saline. On day 29, Morris water maze test was conducted; mice were euthanized, and brain cortex/hippocampus were dissected for Aβ quantification via ELISA [1]

In male Sprague-Dawley rats, oral Avagacestat at 10 mg/kg showed an oral bioavailability of ~45%, a plasma elimination half-life (t₁/₂) of ~3.8 hours, a peak plasma concentration (Cmax) of 320 ng/mL (reached at 1.2 hours post-dose), and a volume of distribution (Vd) of ~2.0 L/kg [1]
- In beagle dogs, oral Avagacestat at 0.5 mg/kg had a t₁/₂ of ~4.5 hours, a Cmax of 45 ng/mL (reached at 1.5 hours post-dose), and a brain-to-plasma concentration ratio of ~0.5 (measured 2 hours post-dose) [4]
- In human AD patients (Phase II trial), oral Avagacestat (100 mg/day) reached a steady-state Cmax of ~180 ng/mL, with a t₁/₂ of ~5.2 hours [5]
- Avagacestat has high plasma protein binding (>98%) in human, rat, and dog plasma (measured via ultrafiltration) [1]

In a 28-day rat toxicity study (oral Avagacestat at 5, 20, 60 mg/kg/day), the no-observed-adverse-effect level (NOAEL) was 20 mg/kg/day; at 60 mg/kg/day, mild skin hyperplasia and gastrointestinal mucosal inflammation were observed in 2/5 rats (reversible after treatment cessation) [1]
- In beagle dogs (0.5 mg/kg oral for 14 days), no significant changes in serum ALT, AST, creatinine, or BUN levels were observed; no histopathological abnormalities in brain, liver, or kidney [4]
- In the Phase II AD trial (n=252), the most common adverse events (AEs) with Avagacestat were diarrhea (18% [50 mg] vs. 25% [100 mg] vs. 8% placebo), fatigue (15% vs. 19% vs. 10%), and skin rash (12% vs. 18% vs. 5%); 5% of 100 mg/day patients discontinued due to AEs [5]

[1]. Letter Discovery and Evaluation of BMS-708163, a Potent, Selective and Orally Bioavailable γ-Secretase Inhibitor. Med Chem Lett, 2010, 1 (3), 120-124.

[2]. γ Secretase inhibitor BMS-708163 reverses resistance to EGFR inhibitor via the PI3K/Akt pathway in lung cancer. J Cell Biochem. 2015 Jun;116(6):1019-27.

[3]. BMS-708,163 targets presenilin and lacks notch-sparing activity. Biochemistry. 2012 Sep 18;51(37):7209-11.

[4]. A canine model to evaluate efficacy and safety of γ-secretase inhibitors and modulators. J Alzheimers Dis. 2012;28(4):809-22.

[5]. Safety and tolerability of the γ-secretase inhibitor avagacestat in a phase 2 study of mild to moderate Alzheimer disease. Arch Neurol. 2012 Nov;69(11):1430-40.

Avagacestat is an oxadiazole and a ring assembly.
Avagacestat has been investigated for the basic science and treatment of Alzheimer Disease.

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Avagacestat (BMS-708163) is a small-molecule γ-secretase inhibitor initially developed for Alzheimer’s disease (AD) (via reducing Aβ production) and later investigated for EGFR inhibitor-resistant NSCLC (via reversing PI3K/Akt-mediated resistance) [1,2]
- Unlike some γ-secretase inhibitors, Avagacestat directly targets the presenilin subunit of γ-secretase, ensuring potent inhibition of Aβ production; however, its lack of Notch-sparing activity increases the risk of Notch-related AEs (e.g., skin rash, gastrointestinal toxicity) [3,5]
- Avagacestat completed Phase II clinical trials for mild-to-moderate AD but was discontinued due to lack of cognitive improvement and increased AEs at higher doses; it also showed preclinical efficacy in EGFR-resistant NSCLC but did not advance to late-stage clinical trials [2,5]
- In canine models, Avagacestat is used as a tool to evaluate γ-secretase inhibitor efficacy in large animals, supporting translational research for AD therapeutics [4]

C20H17CLF4N4O4S

520.88

520.059

C, 46.12; H, 3.29; Cl, 6.81; F, 14.59; N, 10.76; O, 12.29; S, 6.16

1146699-66-2

46883536

White to off-white solid powder

1.5±0.1 g/cm3

652.3±65.0 °C at 760 mmHg

348.3±34.3 °C

0.0±2.0 mmHg at 25°C

1.564

4.89

127.77

1

11

9

34

792

1

ClC1C([H])=C([H])C(=C([H])C=1[H])S(N(C([H])([H])C1C([H])=C([H])C(C2=NOC([H])=N2)=C([H])C=1F)[C@@]([H])(C(N([H])[H])=O)C([H])([H])C([H])([H])C(F)(F)F)(=O)=O

XEAOPVUAMONVLA-QGZVFWFLSA-N

InChI=1S/C20H17ClF4N4O4S/c21-14-3-5-15(6-4-14)34(31,32)29(17(18(26)30)7-8-20(23,24)25)10-13-2-1-12(9-16(13)22)19-27-11-33-28-19/h1-6,9,11,17H,7-8,10H2,(H2,26,30)/t17-/m1/s1

(2R)-2-[(4-chlorophenyl)sulfonyl-[[2-fluoro-4-(1,2,4-oxadiazol-3-yl)phenyl]methyl]amino]-5,5,5-trifluoropentanamide

BMS-708163; BMS708163; Avagacestat; BMS 708163

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 3 mg/mL (5.76 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 30.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。