p38 (IC50 = 50 nM); p38β2 (IC50 = 500 nM)

SB203580 的 IC50 为 3-5 μm，当 IL-2 存在时，可阻断原代人 T 细胞、鼠 CT6 T 细胞或 BAF F7 B 细胞的增殖。尽管所需浓度稍高且 IC50 高于 10 μM，SB203580 也能抑制 IL-2 诱导的 p70S6 激酶激活。 SB203580 的 IC50 范围为 3-10 μM，还以剂量依赖性方式抑制 PDK1 的活性。 p38-MAPK 对 MAPKAPK2 的刺激被 SB203580 抑制，IC50 约为 0.07 μM，而总 SAPK/JNK 活性受到抑制，IC50 为 3-10 μM。较高浓度的 SB203580 会激活 ERK 通路，从而提高 NF-κB 的转录活性。 SB203580 诱导人肝细胞癌细胞 (HCC) 发生自噬。

SB203580 可保护体内模型中的猪心肌免受缺血性损伤。在系统性红斑狼疮 (SLE) 的 MRL/lpr 小鼠模型中，SB203580 可有效预防和治疗该疾病。

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SB203580治疗的MRL/lpr小鼠可预防蛋白尿。[4]
SB203580对MRL/lpr小鼠的ALT和AST没有影响。[4]
SB203580治疗的MRL/lpr小鼠的BUN降低，但Cr没有降低。 SB203580治疗的MRL/lpr小鼠的肾脏重量减少，但脾脏重量没有减少。[4]
SB203580治疗的MRL/lpr小鼠的肾脏病理变化减弱。[4]
SB203580治疗的MRL/lpr小鼠的肝脏病理变化得到缓解。[4]
SB203580治疗的MRL/lpr小鼠的脾脏病理变化得到缓解。[4]
SB203580治疗的MRL/lpr小鼠肾小球IgG、IgM、IgA和C3沉积减少。[4]

将 4 μg 羊抗 PKBα 固定在 25 μL 蛋白 G-Sepharose 上过夜（或 1.5 小时），并在缓冲液 A（50 mm Tris，pH 7.5，1 mm EDTA，1 mm EGTA，0.5 mm Na3VO4，0.1%）中洗涤β-巯基乙醇、1% Triton X-100、50 mm 氟化钠、5 mm 焦磷酸钠、0.1 mm 苯甲基磺酰氟、1 μg/mL 抑肽酶、胃酶抑素、亮肽素和 1 μm 微囊藻毒素）。然后将固定化的抗 PKB 与 0.5 ml 裂解液（来自 5 × 106 个细胞）一起孵育 1.5 小时，并在补充有 0.5 m NaCl 的 0.5 mL 缓冲液 A 中洗涤 3 次，在 0.5 mL 缓冲液 B（50 mm Tris-HCl，pH 7.5，0.03% (w/v) Brij-35，0.1 mm EGTA 和 0.1% β-巯基乙醇），并用 100 μl 测定稀释缓冲液两次； 5× 测定稀释缓冲液为 100 mm MOPS，pH 7.2，125 mm β-甘油磷酸盐，25 mm EGTA，5 mm 原钒酸钠，5 mm DTT。向 PKB 酶免疫复合物中添加 10 μL 测定稀释缓冲液、40 μm 蛋白激酶 A 抑制肽、100 μm PKB 特异性底物肽和 10 μCi 的 [γ-32P]ATP，所有这些均在测定稀释缓冲液中配制。反应在室温下振荡孵育 20 分钟，然后脉冲旋转样品，将 40 μL 反应体积移至另一管中，向其中添加 20 μL 40% 三氯乙酸以终止反应。将其混合并在室温下孵育 5 分钟，然后将 40 μL 转移到 P81 磷酸纤维素纸上并结合 30 秒。 P81 片在室温下用 0.75% 磷酸洗涤 3 次，然后用丙酮洗涤。然后通过闪烁计数测量γ-32P的掺入。

在没有生长因子、抗生素或β-巯基乙醇补充剂的情况下，CT6细胞和BA/F3 F7细胞在RPMI中洗涤3次并在含有5%胎牛血清的RPMI中培养过夜来休息。如图例所示，在 2 mL RPMI、5% 胎牛血清中对 2-5 106 个静息 CT6 细胞与 SB203580 或载体对照进行预孵育。在 37°C 下用 20 ng/ml 重组人 IL-2 刺激孵育 5 分钟后，将细胞在微型离心机中沉淀 30 秒，吸出培养基，并在适当的缓冲液中裂解沉淀。 BA/F3 细胞维持在含有谷氨酰胺的 RPMI 中，另外补充有 5% 胎牛血清和 0.2 μg/mL G418，并稳定表达 IL-2 β 受体链的缺失突变体。然后彻底清洗细胞，静置过夜，然后再次清洗，然后用 IL-2 激活。此类细胞制剂含有 >90% T 细胞。在细胞增殖测定中测量[3H]胸苷的掺入。

Six-week-old female atymic Nu/Nu mice CAL27 p38WT and p38TM tumors[1]
5 mg/kg/day
Intra peritoneal injected daily for 16 consecutive days

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Systemic lupus erythematosus (SLE) are established in female MRL/lpr mice and female C57BL/6 mice
0.1 M/day
Orally administered

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Female MRL/lpr mice were randomized into two groups (n = 10 per group) and were fed control diet (named as group 2 in the following) or diet with SB203580 (named as group 3 in the following) starting at the age of 14 weeks and continuing for up to 22 weeks. Adezmapimod (SB203580) was dissolved in drinking water (250 μmol/L), was orally administered (0.4 ml/day). Ten C57BL/6 female mice were used as negative controls (named as group 1 in the following). Two mice in MRL/lpr group 2 were dead at 16 weeks and 18 weeks of age respectively. Two mice in MRL/lpr group 3 were dead at 19 weeks of age. Significant increase of urine protein (300–2000 mg/dl) was found in each mouse before death, indicating a probable renal failure be the cause of death. Ultimately, 10 mice in group 1, 8 mice in group 2 and group 3 were included in statistical analysis.[4]

[1]. Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J. 2000 Oct 1;351(Pt 1):95-105.

[2]. The pyridinyl imidazole inhibitor SB203580 blocks phosphoinositide-dependent protein kinase activity, protein kinase B phosphorylation, and retinoblastoma hyperphosphorylation in interleukin-2-stimulated T cells independently of p38 mitogen-activated protein kinase. J Biol Chem. 2000 Mar 10;275(10):7395-402.

[3]. A role for p38 MAPK in head and neck cancer cell growth and tumor-induced angiogenesis and lymphangiogenesis. Mol Oncol. 2014 Feb;8(1):105-18.

[4]. The selective p38 mitogen-activated protein kinase inhibitor, SB203580, improves renal disease in MRL/lpr mouse model of systemic lupus. Int Immunopharmacol. 2011 Sep;11(9):1319-26.

Pyridinyl imidazole inhibitors, particularly SB203580, have been widely used to elucidate the roles of p38 mitogen-activated protein (MAP) kinase (p38/HOG/SAPKII) in a wide array of biological systems. Studies by this group and others have shown that SB203580 can have antiproliferative activity on cytokine-activated lymphocytes. However, we recently reported that the antiproliferative effects of SB203580 were unrelated to p38 MAP kinase activity. This present study now shows that SB203580 can inhibit the key cell cycle event of retinoblastoma protein phosphorylation in interleukin-2-stimulated T cells. Studies on the proximal regulator of this event, the phosphatidylinositol 3-kinase/protein kinase B (PKB)(Akt/Rac) kinase pathway, showed that SB203580 blocked the phosphorylation and activation of PKB by inhibiting the PKB kinase, phosphoinositide-dependent protein kinase 1. The concentrations of SB203580 required to block PKB phosphorylation (IC(50) 3-5 microM) are only approximately 10-fold higher than those required to inhibit p38 MAP kinase (IC(50) 0.3-0.5 microM). These data define a new activity for this drug and would suggest that extreme caution should be taken when interpreting data where SB203580 has been used at concentrations above 1-2 microM.[2]

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We have recently gained a remarkable understanding of the mutational landscape of head and neck squamous cell carcinoma (HNSCC). However, the nature of the dysregulated signaling networks contributing to HNSCC progression is still poorly defined. Here, we have focused on the role of the family of mitogen activated kinases (MAPKs), extracellular regulated kinase (ERK), c-Jun terminal kinase (JNK) and p38 MAPK in HNSCC. Immunohistochemical analysis of a large collection of human HNSCC tissues revealed that the levels of the phosphorylated active form of ERK1/2 and JNK were elevated in less than 33% and 16% of the cases, respectively. Strikingly, however, high levels of active phospho-p38 were observed in most (79%) of hundreds of tissues analyzed. We explored the biological role of p38 in HNSCC cell lines using three independent approaches: treatment with a specific p38 inhibitor, SB203580; a retro-inhibition strategy consisting in the use of SB203580 combined with the expression of an inhibitor-insensitive mutant form of p38α; and short-hairpin RNAs (shRNAs) targeting p38α. We found that specific blockade of p38 signaling significantly inhibited the proliferation of HNSCC cells both in vitro and in vivo. Indeed, we observed that p38 inhibition in HNSCC cancer cells reduces cancer growth in tumor xenografts and a remarkable decrease in intratumoral blood and lymphatic vessels. We conclude that p38α functions as a positive regulator of HNSCC in the context of the tumor microenvironment, controlling cancer cell growth as well as tumor-induced angiogenesis and lymphangiogenesis.[3]

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Systemic lupus erythematosus (SLE) is an autoimmune disease accompanying excessive inflammatory responses in a wide range of organs. Abnormal activation of p38 MAPK has been postulated to contribute to the inflammation of SLE, leading to progressive tissue and organ damages to develop lupus nephritis and autoimmune hepatitis. In order to determine whether p38 MAPK inhibitor is effective in mouse model of SLE, a specific inhibitor of p38 MAPK SB203580 was orally administrated to MRL/lpr mice aged from 14 to 22 weeks. Renal and hepatic functions, as well as pathologic changes of important organs including kidney, liver and spleen of MRL/lpr mice were evaluated. As a result, we showed that SB203580 improved renal function by decreasing the levels of proteinuria and serum BUN, ameliorating the pathologic changes of kidney and reducing Ig and C(3) depositions in the kidney. Hepatocytes necrosis, recruitment and proliferation of leucocytes in liver and spleen were found to be inhibited by administration of SB203580. Therefore, p38 MAPK activation may be partially responsible for escalating autoimmune renal, hepatic and splenic destruction, and its inhibitor may lighten the autoimmune attack in these important organs and improve renal function. Our study reveals that the selective blockade of p38 MAPK is effective to prevent and treat the disease in this model of SLE.[4]

C₂₁H₁₇CLFN₃OS

413.90

413.076

C, 60.94; H, 4.14; Cl, 8.56; F, 4.59; N, 10.15; O, 3.87; S, 7.75

869185-85-3

Adezmapimod;152121-47-6

16760644

Light yellow to yellow solid powder

6.349

77.85

2

5

4

28

500

0

Cl.O=S(C)C1C=CC(C2NC(C3C=CC(F)=CC=3)=C(C3C=CN=CC=3)N=2)=CC=1

WOSGGXINSLMASH-UHFFFAOYSA-N

InChI=1S/C21H16FN3OS.ClH/c1-27(26)18-8-4-16(5-9-18)21-24-19(14-2-6-17(22)7-3-14)20(25-21)15-10-12-23-13-11-15;/h2-13H,1H3,(H,24,25);1H

4-[4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-1H-imidazol-5-yl]pyridine;hydrochloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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 (6.04 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 (6.04 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: 4% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/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。