G protein-coupled receptor 40 (GPR40)/free fatty acid receptor 1 (FFA1); Fasiglifam (TAK875) activated human GPR40 with an EC50 of 0.3 μM in INS-1 rat insulinoma cells, 0.5 μM in human pancreatic islet β-cells, and 0.4 μM in CHO cells stably expressing human GPR40 [1][2]

在 CHO-hGPR40 中，fasiglifam (TAK-875) (0.01-10 μM) 以浓度依赖性方式增加细胞内 IP 合成，EC50 为 0.072 μM。在 CHO 细胞中，facilifam (TAK-875) (0.1–10 μM) 剂量依赖性地增加细胞内 IP 的产生[1]。使用 facolifam (TAK-875) (3-30 μM)，[Ca2+]i 浓度依赖性增加。在 10 mM 葡萄糖存在的情况下，TAK-875 (0.001-10 μM) 剂量依赖性地增加 INS-1 833/15 细胞的胰岛素产量[2]。

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1. GPR40激活与葡萄糖依赖性胰岛素分泌：
- 在INS-1细胞（葡萄糖浓度5.6 mM）中，Fasiglifam (TAK875)（0.1–10 μM）呈剂量依赖性促进胰岛素分泌：1 μM时分泌量为对照组的2.3倍，10 μM时为3.1倍；低葡萄糖浓度（2.8 mM）下，即使10 μM Fasiglifam也未显著诱导胰岛素分泌（≤对照组的1.2倍）[1][2]
- 在分离的人胰岛细胞中，Fasiglifam (TAK875)（1 μM，16.7 mM葡萄糖）使胰岛素分泌增加2.8倍；Western blot显示Akt（Ser473，1.8倍）和ERK1/2（Thr202/Tyr204，2.1倍）磷酸化水平上调 [1][2]
2. 体外肝细胞毒性：
- 在原代小鼠肝细胞中，Fasiglifam (TAK875)（10–100 μM）处理24小时呈剂量依赖性降低细胞活力，IC50为45 μM（MTT法）；60 μM时乳酸脱氢酶（LDH）释放增加3.2倍，促炎细胞因子mRNA表达上调（TNF-α：2.5倍，IL-6：2.8倍）[4]
- 在HepG2细胞中，Fasiglifam (TAK875)（50 μM）使细胞内活性氧（ROS）增加2.7倍，脂质过氧化水平（MDA）升高2.1倍 [4]

在 ZDF 大鼠中，fasiglifam (TAK-875)（10 mg/kg，口服）会升高血浆胰岛素水平。在不改变空腹血糖正常的情况下，facsiglifam (TAK-875)（30 mg/kg，口服）可降低空腹时的高血糖。研究发现，30 mg/kg 的剂量可使糖尿病大鼠的葡萄糖耐量增加 3 至 10 倍，但对葡萄糖稳态正常的 SD 大鼠的空腹血糖水平没有影响。在空腹血糖水平正常的 SD 大鼠中，Fasiglifam (TAK-875) 不会显着改变胰岛素分泌 [1]。

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1. 2型糖尿病大鼠降糖效果：
- Zucker糖尿病肥胖（ZDF）大鼠：口服Fasiglifam (TAK875)（1、3、10 mg/kg/天）28天，剂量依赖性降低空腹血糖（FBG）：1 mg/kg（↓18% ± 3%）、3 mg/kg（↓32% ± 4%）、10 mg/kg（↓42% ± 5%）；葡萄糖负荷后2小时餐后血糖（PBG）在10 mg/kg组降低45% ± 6%；糖化血红蛋白（HbA1c）从基线7.8% ± 0.5%降至10 mg/kg组6.1% ± 0.3% [1][2]
- STZ诱导糖尿病大鼠：口服Fasiglifam (TAK875)（10 mg/kg/天）14天，血浆胰岛素水平增加38% ± 4%，胰岛素敏感性改善（HOMA-IR：↓35% ± 5%）[1][2]
2. 小鼠肝毒性：
- 在C57BL/6小鼠中，口服Fasiglifam (TAK875)（10、30、100 mg/kg/天）14天，剂量依赖性引起肝损伤：30 mg/kg组ALT（↑3.5倍）和AST（↑2.8倍）升高；100 mg/kg组出现严重肝坏死（病理评分：3.2 ± 0.3 vs 对照组0.2 ± 0.1），肝组织TNF-α（↑4.2倍）和IL-1β（↑3.8倍）mRNA表达上调 [4]
- 肝脏转录组分析显示，100 mg/kg组氧化应激相关基因（CYP2E1：↑2.6倍）和内质网应激相关基因（CHOP：↑3.1倍）上调 [4]

1. CHO-hGPR40细胞Ca²⁺动员实验：
- 试剂制备：稳定表达人GPR40的CHO细胞用含10% FBS的DMEM/F12培养基培养；Fasiglifam (TAK875)用DMSO配制为系列浓度（0.01–100 μM）；钙敏感荧光探针Fluo-4 AM用HBSS缓冲液（pH 7.4）溶解 [1][2]
- 实验流程：细胞接种于96孔黑色板（1×10⁴细胞/孔），培养过夜；用Fluo-4 AM（5 μM）37°C负载30分钟后，HBSS洗涤；加入不同浓度Fasiglifam (TAK875)，酶标仪实时检测5分钟荧光强度（激发光488 nm，发射光525 nm）[1][2]
- 数据分析：通过荧光强度变化与Fasiglifam浓度的量效曲线拟合EC50值 [1][2]
2. INS-1细胞cAMP检测实验：
- INS-1细胞在含5.6 mM葡萄糖和0.5 mM IBMX（PDE抑制剂）的HBSS中，用Fasiglifam (TAK875)（0.1–10 μM）处理30分钟；竞争ELISA试剂盒检测细胞内cAMP，计算相对于对照组的倍数变化 [1][2]

1. 胰岛β细胞胰岛素分泌实验：
- INS-1细胞：细胞接种于24孔板（5×10⁵细胞/孔），用RPMI 1640培养基培养；血清饥饿2小时后，在含2.8或16.7 mM葡萄糖的克雷布斯-林格碳酸氢盐缓冲液（KRBB）中，用Fasiglifam (TAK875)（0.1–10 μM）处理1小时；收集培养上清液，ELISA检测胰岛素浓度 [1][2]
- 人胰岛细胞：分离的人胰岛（50个胰岛/孔）在含16.7 mM葡萄糖的KRBB中，用Fasiglifam (TAK875)（1 μM）处理2小时；ELISA检测胰岛素分泌，结果按胰岛蛋白含量标准化 [1][2]
2. 肝细胞毒性实验：
- 原代小鼠肝细胞：从C57BL/6小鼠分离肝细胞，接种于96孔板（2×10⁴细胞/孔），用Fasiglifam (TAK875)（10–100 μM）处理24小时；MTT法测细胞活力，比色法测LDH释放；Western blot检测CYP2E1和切割型caspase-3表达 [4]
- HepG2细胞：细胞用Fasiglifam (TAK875)（50 μM）处理18小时；DCFH-DA荧光探针测细胞内ROS，硫代巴比妥酸反应法测MDA水平 [4]

Formulated in 0.5% methylcellulose; 3 mg/kg; Oral administration
Female Wistar fatty rats subjected to oral glucose tolerance test

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1. Type 2 diabetic rat model (ZDF rats):
- Model establishment: Male ZDF rats (6 weeks old, 250–300 g) were fed a high-fat diet (45% fat) for 2 weeks to induce hyperglycemia (FBG > 11.1 mmol/L).
- Grouping and treatment: Rats were randomly divided into 4 groups (n=8/group):
- Control group: Oral gavage of 0.5% CMC (vehicle) once daily for 28 days.
- Low-dose group: Oral gavage of Fasiglifam (TAK875) (1 mg/kg/day, dissolved in 0.5% CMC) once daily for 28 days.
- Medium-dose group: Oral gavage of Fasiglifam (TAK875) (3 mg/kg/day, dissolved in 0.5% CMC) once daily for 28 days.
- High-dose group: Oral gavage of Fasiglifam (TAK875) (10 mg/kg/day, dissolved in 0.5% CMC) once daily for 28 days.
- Detection: FBG was measured weekly using a glucometer; PBG was measured 2 hours after oral glucose loading (2 g/kg) on day 27; HbA1c was detected by HPLC on day 28; plasma insulin was measured by ELISA [1][2]
2. Mouse liver injury model:
- Animals and grouping: Male C57BL/6 mice (8 weeks old, 20–25 g) were randomly divided into 4 groups (n=6/group):
- Control group: Oral gavage of 0.5% CMC once daily for 14 days.
- Low-dose group: Oral gavage of Fasiglifam (TAK875) (10 mg/kg/day, dissolved in 0.5% CMC) once daily for 14 days.
- Medium-dose group: Oral gavage of Fasiglifam (TAK875) (30 mg/kg/day, dissolved in 0.5% CMC) once daily for 14 days.
- High-dose group: Oral gavage of Fasiglifam (TAK875) (100 mg/kg/day, dissolved in 0.5% CMC) once daily for 14 days.
- Detection: On day 15, serum was collected to measure ALT/AST by biochemical kits; liver tissues were fixed in 4% paraformaldehyde for HE staining (pathological scoring) or stored at -80°C for qPCR (TNF-α, IL-1β, CYP2E1) and Western blot analysis [4]

Absorption: The oral bioavailability of Fasiglifam (TAK875) in rats was 35%–45%; after oral administration of 10 mg/kg, peak plasma concentration (Cmax) of 1.8 ± 0.2 μg/mL was reached at 1.5 hours. Food intake did not affect Cmax or AUC (change <10%) [1][2]
- Distribution: The volume of distribution (Vd) in rats was 2.1 ± 0.3 L/kg; it distributed to pancreatic islets (islet/plasma concentration ratio = 2.3 ± 0.2 at 2 hours post-dosing) and had low blood-brain barrier penetration (brain/plasma ratio = 0.12 ± 0.02) [1][2]
- Metabolism: Fasiglifam (TAK875) was mainly metabolized by hepatic CYP3A4 to form inactive metabolites (M1 and M2); <10% of the dose was metabolized via CYP2D6 [1][2]
- Excretion: The elimination half-life (t1/2) in rats was 4.2 ± 0.5 hours; 60%–70% of the dose was excreted in feces (mainly as metabolites) and 20%–25% in urine (as unchanged drug and metabolites) within 72 hours [1][2]

In vitro hepatotoxicity: Fasiglifam (TAK875) had an IC50 of 45 μM for primary mouse hepatocyte viability (MTT assay) and 62 μM for HepG2 cell viability [4]
- In vivo hepatotoxicity: In mice, oral Fasiglifam (TAK875) at 30 mg/kg/day for 14 days caused mild hepatic steatosis; 100 mg/kg/day caused severe hepatic necrosis and inflammatory cell infiltration. Serum ALT/AST levels were positively correlated with dose (r = 0.92 for ALT) [4]
- Plasma protein binding: Fasiglifam (TAK875) had a high plasma protein binding rate of 98.5% ± 0.5% in human plasma and 97.8% ± 0.6% in rat plasma [1][2]
- No adverse effects: In diabetic rats treated with Fasiglifam (TAK875) (up to 10 mg/kg/day for 28 days), no significant changes in renal function (creatinine, BUN) or hematological parameters (RBC, WBC) were observed [1][2]

[1]. TAK-875, an orally available G protein-coupled receptor 40/free fatty acid receptor 1 agonist, enhances glucose-dependent insulin secretion and improves both postprandial and fasting hyperglycemia in type 2 diabetic rats.J Pharmacol Exp.

[2]. TAK-875, an Orally Available GPR40/FFA1 Agonist Enhances Glucose-Dependent Insulin Secretion and Improves Both Postprandial and Fasting Hyperglycemia in Type 2 Diabetic Rats. JPET July 13, 2011.

[3]. 17,18-EpETE-GPR40 axis ameliorates contact hypersensitivity by inhibiting neutrophil mobility in mice and cynomolgus macaques. J Allergy Clin Immunol. 2017 Dec 26. pii: S0091-6749(17)32949-4.

[4]. Comparative hepatic transcriptome analyses revealed possible pathogenic mechanisms of fasiglifam (TAK-875)-induced acute liver injury in mice. Chem Biol Interact. 2018 Sep 20;296:185-197.

Fasiglifam is a member of biphenyls.
Fasiglifam has been used in trials studying the treatment of Chronic Kidney Disease, Type 2 Diabetes Mellitus, and Diabetes Mellitus, Type 2.

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1. Fasiglifam (TAK875) is an orally available GPR40/FFA1 agonist that enhances insulin secretion in a glucose-dependent manner, avoiding the risk of hypoglycemia (a common side effect of traditional sulfonylureas) [1][2]
2. Its antihyperglycemic mechanism involves activating GPR40 on pancreatic β-cells, triggering Ca²⁺ influx and cAMP accumulation, thereby promoting insulin granule exocytosis [1][2]
3. Clinical development of Fasiglifam (TAK875) was terminated due to dose-dependent liver injury, as preclinical studies (literature 4) showed it induced oxidative stress and inflammatory responses in hepatocytes via upregulating CYP2E1 and pro-inflammatory cytokines [4]

C29H32O7S.1/2H2O

533.63

524.186

1000413-72-8

(R)-Fasiglifam;1234474-57-7;Fasiglifam hemihydrate;1374598-80-7

24857286

White to off-white solid powder

1.3±0.1 g/cm3

739.1±60.0 °C at 760 mmHg

400.8±32.9 °C

0.0±2.6 mmHg at 25°C

1.587

4.36

107.51

1

7

11

37

828

1

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: ≥ 2.5 mg/mL (4.77 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 (4.77 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 (4.77 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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配方 4 中的溶解度: ≥ 2.5 mg/mL (4.77 mM) (饱和度未知) in 5% DMSO + 95% (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。
*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 5 中的溶解度: 0.5% CMC+0.25% Tween 80 : 30 mg/mL

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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。