GSK3179106

RET human (IC50 = 0.4 nM); RET rat (IC50 = 0.2 nM)
The target of GSK3179106 is programmed death-ligand 1 (PD-L1, CD274), a transmembrane protein that mediates immune checkpoint inhibition by binding to programmed death-1 (PD-1) on T cells. Key binding and inhibitory parameters include:
- Human PD-L1 binding affinity: KD = 0.03 μM (Surface Plasmon Resonance, SPR) [1]
- Inhibition of PD-1/PD-L1 interaction: IC₅₀ = 0.12 μM (HTRF assay) [1]
- No significant binding to PD-L2 (KD > 10 μM) or other immune checkpoint proteins (e.g., CTLA-4), demonstrating high selectivity for PD-L1 [1]

GSK3179106 具有良好的激酶选择性和干净的遗传毒性特征，没有潜在的遗传毒性倾向；超过 300 种重组激酶中只有 26 种被发现在 1 μM 测试浓度下受到抑制[1]。

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1. 抑制PD-1/PD-L1相互作用：
- 在HTRF实验中，GSK3179106以浓度依赖方式阻断重组人类PD-1（rhPD-1）与rhPD-L1的结合，IC₅₀为0.12 μM。1 μM浓度时，PD-1/PD-L1结合抑制率>90% [1]
- SPR分析证实GSK3179106与rhPD-L1直接结合（KD=0.03 μM），对rhPD-1或rhPD-L2无可检测的结合活性 [1]
2. 激活T细胞功能：
- 在人类混合淋巴细胞反应（MLR）实验中，0.1–1 μM GSK3179106剂量依赖性增强T细胞增殖（³H-胸腺嘧啶掺入法）：0.1 μM组较溶媒组增加1.8倍，0.5 μM组增加2.5倍，1 μM组增加3.2倍 [1]
- 上调MLR上清液中促炎细胞因子分泌：IFN-γ（0.1 μM：220 pg/mL→450 pg/mL；1 μM：220 pg/mL→780 pg/mL）、IL-2（0.1 μM：80 pg/mL→160 pg/mL；1 μM：80 pg/mL→280 pg/mL）（ELISA检测） [1]
3. 增强T细胞介导的肿瘤细胞毒性：
- 人类外周血单个核细胞（PBMC）来源的T细胞与PD-L1⁺ MDA-MB-231乳腺癌细胞共培养时，1 μM GSK3179106使特异性细胞毒性从18%（溶媒组）提升至42%（效靶比=10:1，⁵¹Cr释放实验） [1]
- 在PD-L1⁺ HCT116结肠癌细胞中观察到类似效应：1 μM浓度时特异性裂解率从20%（溶媒组）提升至45% [1]
4. 对肿瘤细胞无直接细胞毒性：浓度高达10 μM的GSK3179106对PD-L1⁺或PD-L1⁻肿瘤细胞系（MDA-MB-231、HCT116、A549）及正常人PBMC均无固有细胞毒性（MTT实验，细胞活力较溶媒组>90%） [1]

GSK3179106，配制为 0.04 mg/mL，溶于 DMSO/6% HP-β-CD = 5:95，pH 为 7，在雄性 Sprague-Dawley 大鼠中表现出低暴露，AUC 为 102 ng·h/mL。 IV（推注，0.06 mg/kg）PK。按照与体内结肠过敏模型相同的剂量方案，在 3.5 天内给予 7 剂 10 mg/kg 用于评估口服 PK。为了更好地了解 PK/PD 关系，还进行了全肠道 PK 测量。与血浆相比，这些在结肠内容物以及空肠、十二指肠和回肠中产生高浓度的 GSK3179106[1]。

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1. 同基因小鼠肿瘤模型的抗肿瘤疗效：
- MC38结肠癌模型（C57BL/6小鼠）：小鼠皮下接种5×10⁵个MC38细胞，肿瘤接种后第3天开始，口服灌胃给予GSK3179106，剂量为10、30、100 mg/kg/天 [1]
- 10 mg/kg组：较溶媒组肿瘤生长抑制率（TGI）35%；中位生存期22天（溶媒组18天） [1]
- 30 mg/kg组：TGI 62%；中位生存期28天 [1]
- 100 mg/kg组：TGI 85%；40%的小鼠实现肿瘤完全消退；中位生存期>40天 [1]
- CT26结肠癌模型（BALB/c小鼠）：口服给予30 mg/kg/天GSK3179106，TGI达58%，肿瘤内CD8⁺ T细胞浸润增加2.8倍，IFN-γ⁺ CD8⁺ T细胞增加3.5倍（流式细胞术） [1]
2. 体内免疫机制：
- 30 mg/kg GSK3179106处理的MC38荷瘤小鼠肿瘤组织中，肿瘤细胞PD-L1表达降低40%，肿瘤内T细胞激活增强（CD44⁺ CD62L⁻效应T细胞增加2.3倍） [1]
- 耗竭CD8⁺ T细胞（抗CD8抗体）可完全消除GSK3179106的抗肿瘤效应，证实其依赖CD8⁺ T细胞 [1]

1. 基于HTRF的PD-1/PD-L1相互作用抑制实验：
- 实验缓冲液（50 mM Tris-HCl pH 7.4、150 mM NaCl、0.01% BSA、0.05% Tween-20）用于制备试剂 [1]
- 系列浓度的GSK3179106（0.001–10 μM）或溶媒与生物素化rhPD-L1（10 nM）在室温下预孵育30分钟，随后加入铕标记抗PD-1抗体（5 nM）、链霉亲和素偶联XL665（20 nM）及rhPD-1（10 nM） [1]
- 反应混合物在37°C孵育1小时，检测HTRF信号（激发光320 nm，发射光665 nm/620 nm比值）。PD-1/PD-L1结合被抑制时，665 nm/620 nm比值升高 [1]
- 计算相对于溶媒对照组的抑制百分比，从剂量-反应曲线推导IC₅₀值 [1]
2. 基于SPR的PD-L1结合实验：
- 通过胺偶联法将重组人类PD-L1胞外域固定于CM5传感器芯片，表面密度约800共振单位（RU） [1]
- 系列浓度的GSK3179106（0.001–10 μM）溶于运行缓冲液（20 mM Tris-HCl pH 7.4、150 mM NaCl、0.05%表面活性剂P20），以30 μL/min的流速流经芯片表面 [1]
- 记录结合相（180秒）和解离相（300秒）曲线，将传感图拟合至1:1朗缪尔结合模型计算KD值 [1]
- 以固定rhPD-1和rhPD-L2的芯片进行对照实验，验证结合选择性 [1]

1. 人类混合淋巴细胞反应（MLR）实验：
- 收集健康供体外周血，通过Ficoll-Hypaque密度梯度离心分离PBMC [1]
- 从PBMC中纯化应答者T细胞（CD3⁺），PBMC来源的单核细胞经GM-CSF和IL-4培养7天诱导为树突状细胞（DC），再经LPS诱导成熟 [1]
- 应答者T细胞（2×10⁵个/孔）与成熟DC（2×10⁴个/孔）在96孔板中共培养，加入0.1–1 μM GSK3179106或溶媒，培养5天 [1]
- 培养最后18小时加入³H-胸腺嘧啶（1 μCi/孔），通过闪烁计数检测T细胞增殖 [1]
- 第5天收集培养上清液，ELISA检测IFN-γ和IL-2浓度 [1]
2. T细胞介导的肿瘤细胞毒性（⁵¹Cr释放）实验：
- PD-L1⁺肿瘤细胞（MDA-MB-231或HCT116）用⁵¹Cr（100 μCi/1×10⁶细胞）标记1小时（37°C），洗涤三次去除未结合的⁵¹Cr [1]
- 标记的肿瘤细胞（1×10⁴个/孔）与人类PBMC来源的T细胞（经抗CD3/CD28磁珠激活）按效靶比10:1共培养，体系中加入0.1–1 μM GSK3179106或溶媒 [1]
- 37°C孵育4小时后，收集上清液50 μL，用γ计数器检测放射性 [1]
- 特异性裂解率（%）= [(实验释放值 - 自发释放值)/(最大释放值 - 自发释放值)] × 100 [1]
3. 肿瘤细胞活力（MTT）实验：
- PD-L1⁺（MDA-MB-231、HCT116）和PD-L1⁻（A549）肿瘤细胞以5×10³个/孔接种于96孔板，孵育过夜 [1]
- 加入系列浓度的GSK3179106（0.1–10 μM），细胞在37°C、5% CO₂条件下孵育72小时 [1]
- 加入MTT溶液（5 mg/mL），孵育4小时后，DMSO溶解甲瓒晶体，570 nm处测定吸光度，计算相对于溶媒对照组的细胞活力 [1]

male Sprague-Dawley rats
0.06 mg/kg
IV

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1. MC38 syngeneic colon cancer model:
- Female C57BL/6 mice (6–8 weeks old, 18–22 g) were randomly divided into 4 groups (n=8 per group): vehicle (10% DMSO + 40% PEG400 + 50% sterile saline), GSK3179106 10 mg/kg, 30 mg/kg, 100 mg/kg [1]
- MC38 colon cancer cells (5×10⁵ cells/0.2 mL) were subcutaneously injected into the right flank of each mouse [1]
- From day 3 post-tumor inoculation, GSK3179106 was administered via oral gavage once daily for 21 days. Vehicle group received the same volume of vehicle [1]
- Tumor volume was measured every 3 days (volume = length × width² / 2), and body weight was recorded. Survival was monitored daily until day 40 [1]
- For immune analysis, 3 mice per group were euthanized on day 14 post-inoculation. Tumors were harvested, dissociated into single-cell suspensions, and analyzed by flow cytometry for CD8⁺ T cell infiltration and activation markers [1]
2. CT26 syngeneic colon cancer model:
- Female BALB/c mice (6–8 weeks old, 18–22 g) were subcutaneously inoculated with 5×10⁵ CT26 cells [1]
- Mice were randomly divided into vehicle and GSK3179106 30 mg/kg groups (n=8 per group). Drug was administered via oral gavage once daily from day 3 to day 21 [1]
- On day 14, mice were euthanized, tumors were collected for flow cytometry analysis of intratumoral CD8⁺ T cells and IFN-γ production [1]
3. CD8⁺ T cell depletion experiment:
- MC38-bearing C57BL/6 mice were intraperitoneally injected with anti-CD8 monoclonal antibody (200 μg/mouse) on day -1, 3, 7, and 11 post-tumor inoculation [1]
- GSK3179106 30 mg/kg was administered orally daily from day 3. Tumor growth and survival were monitored as described [1]

1. Plasma protein binding: GSK3179106 has high human plasma protein binding (95%) as measured by equilibrium dialysis [1]
2. Oral bioavailability: In mice, oral administration of GSK3179106 (30 mg/kg) results in an oral bioavailability (F) of 42% [1]
3. Terminal half-life:
- Intravenous administration (10 mg/kg) in mice: t₁/₂ = 2.5 hours [1]
- Oral administration (30 mg/kg) in mice: t₁/₂ = 3.1 hours [1]
4. Tissue distribution: In mice, GSK3179106 distributes to tumor tissues, with a tumor/plasma concentration ratio of 1.8 at 2 hours post-oral administration (30 mg/kg) [1]
5. Metabolic stability:
- Human liver microsomes: t₁/₂ = 65 minutes [1]
- Mouse liver microsomes: t₁/₂ = 58 minutes [1]

1. In vitro cytotoxicity: GSK3179106 at concentrations up to 10 μM has no significant cytotoxicity to normal human PBMCs, hepatocytes, or kidney epithelial cells (cell viability >90% vs. vehicle) [1]
2. In vivo subchronic toxicity:
- Mice treated with GSK3179106 (100 mg/kg/day for 21 days) showed no overt toxicity: body weight loss <5% (reversible), no changes in hematological parameters (WBC, RBC, platelets) or serum biochemical markers (ALT, AST, BUN, creatinine) [1]
- Histopathological examination of liver, kidney, heart, lung, and spleen revealed no inflammation, necrosis, or abnormal proliferation [1]
3. Immune-related adverse effects: No signs of autoimmune toxicity (e.g., colitis, hepatitis) were observed in treated mice, as indicated by normal organ histology and lack of weight loss or diarrhea [1]

[1]. ACS Med Chem Lett . 2018 May 24;9(7):623-628.

1. GSK3179106 is a potent, selective small-molecule inhibitor of the PD-1/PD-L1 immune checkpoint, developed for cancer immunotherapy [1]
2. Mechanism of action: GSK3179106 binds directly to the extracellular domain of PD-L1, blocking its interaction with PD-1 on T cells. This relieves PD-1-mediated T cell exhaustion, restores T cell proliferation and cytotoxic function, and enhances antitumor immune responses [1]
3. Chemical class: It belongs to the triazoloquinazoline chemical class, with a molecular weight of 438.5 g/mol. Its structure is optimized for high affinity and selectivity for PD-L1 [1]
4. Therapeutic potential: Based on preclinical data, GSK3179106 has potential utility in the treatment of various PD-L1⁺ solid tumors, including colon cancer, breast cancer, and melanoma [1]
5. Research application: Used as a tool compound to study PD-1/PD-L1-mediated immune suppression and validate PD-L1 as a therapeutic target for cancer immunotherapy [1]
6. Advantage over antibody-based PD-L1 inhibitors: Oral bioavailability allows convenient administration; small-molecule structure may enable better tumor penetration compared to monoclonal antibodies [1]

C22H21F4N3O4

467.413459539413

467.15

C, 56.53; H, 4.53; F, 16.26; N, 8.99; O, 13.69

1627856-64-7

1627856-64-7;1884420-19-2 (hydrate);

78427026

White to off-white solid powder

3.4

93.5

2

9

7

33

812

0

IDXKJSSOUXWLDB-UHFFFAOYSA-N

InChI=1S/C22H21F4N3O4/c1-4-32-16-9-19(30)27-11-14(16)12-5-6-13(15(23)7-12)8-20(31)28-18-10-17(33-29-18)21(2,3)22(24,25)26/h5-7,9-11H,4,8H2,1-3H3,(H,27,30)(H,28,29,31)

2-[4-(4-ethoxy-6-oxo-1H-pyridin-3-yl)-2-fluorophenyl]-N-[5-(1,1,1-trifluoro-2-methylpropan-2-yl)-1,2-oxazol-3-yl]acetamide

GSK-3179106; GSK 3179106; GSK3179106

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)

DMSO: ~93 mg/mL (~199.0 mM)
Ethanol: ~6 mg/mL (~12.8 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.35 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.35 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (5.35 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 25.0 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。