T-5224
目录号 : GC16165
T-5224 是一种非肽类小分子 AP-1 抑制剂，特异性抑制 AP-1 与启动子区 AP-1 结合位点的结合，最初开发为抗炎药用于治疗类风湿性关节炎 (RA) 抑制炎症细胞因子和 MMPs 而没有任何副作用。
Cas No.:530141-72-1
Sample solution is provided at 25 µL, 10mM.
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Related Biological Data
Knockdown of AP-1 subunits caused the downregulation of TGFB1. (G) The effects of blocking the NF-κB and AP-1 signaling pathways on the expression of TGFB1.
Cells were treated with two NF-κB inhibitors, TPCA1, and BOT-64, and two AP-1 inhibitors, T-5224 (50µM, GLPBIO, Montclair, CA, USA, #GC16165) and SR11302 for 4h and then stimulated with 50ng/mL recombinant IL-1β for 2h.
Int J Biol Sci 16.2 (2020): 204. PMID: 31929749 IF: 6.58 -
Related Biological Data
EML4-ALK fusion protein regulates TF expression through pERK1/2/AP-1 pathway in H2228 cells. (C1-4) Pretreating H2228 cells with AP-1 inhibitor (T-5224, 400μm/L) also attenuated TF expression compared with DMSO control both at RNA (n=6) and protein level (n=3). Data are presented as means±SEM.
H2228 cells were Pretreated with T-5224 (GLPBIO, USA, 400μm/L).
J Thromb Thrombolys, 2023: 1-15. PMID: 37940761 IF: 4
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- Purity: >98.00%
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- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Human oral squamous carcinoma cell lines, OSC-19 and HSC-3-M3. |
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Preparation Method |
Cells were grown in DMEM supplemented with 10% FBS and penicillin (50 units ⁄ mL) ⁄ streptomycin (50 μg ⁄ mL) in a humidified atmosphere (5% CO2) at 37°C. T-5224 was dissolved in DMSO and diluted in culture medium to the target concentration for each experiment. For oral gavage administration, T-5224 was dissolved in polyvinylpyrrolidone (PVP) solution according to the manufacturer’s recommended procedure. |
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Reaction Conditions |
Cells were incubated in T-5224 (0–80 μM), and cell counts were carried out at 24, 48, and 72 h after treatment. |
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Applications |
T-5224 treatment could inhibit the invasion activity of highly metastatic tumor cell line HSC-3-M3. Cell migration was potently inhibited by T-5224 in both cell lines in a dose-dependent manner. Migration activity was almost completely inhibited in the medium containing 80 μM T-5224. |
| Animal experiment [1]: | |
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Animal models |
Female 5–7-week-old BALB⁄ c mice |
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Preparation Method |
HSC-3-M3 (1 x 105 cells) were suspended in 50 μL Hank’s Balanced Salts Solution and injected in the flank of the tongue at day 0. T-5224 was diluted in PVP solution, and T-5224 was given orally to the mice of the treatment group every day from day 1 for 4 weeks. |
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Dosage form |
150 mg⁄ kg |
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Applications |
T-5224 showed significant inhibitory effects against lymph node metastasis in the animal model of HNSCC. The rate of positive metastasis was significantly lower by T-5224 treatment. It is possible that higher doses (150 mg/kg) might be required for cells in which activity of AP-1 is intensively and constitutively activated by genetic mutations or substantial inflammation. |
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References: [1]. Daisuke K. et al. Selective activator protein-1 inhibitor T-5224 prevents lymph node metastasis in an oral cancer model. Cancer Sci. 2016 May; 107 (5) 666–673. |
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T-5224 is a non-peptidic small molecule AP-1 inhibitor that specifically inhibits the binding of AP-1 to the AP-1 binding site of the promoter region and was originally developed as an anti-inflammatory drug for the treatment of rheumatoid arthritis (RA) suppressing inflammatory cytokines and MMPs without any side-effects.[1]
In vitro study indicated that transcriptional inhibition of the expression of MMPs by T-5224 is not limited to tumor cells only; it also occurs in the ECM of the surrounding tissue, and consequently, inhibits tumor cells from infiltrating the surrounding tissue. In addition, T-5224 potently inhibits the essential steps of metastasis, infiltration through the basement membrane barrier and migration into the ECM, by transcriptionally suppressing the expression of MMP-2 and -9.[1]
In vivo experiments demonstrated that T-5224 blocked serum TNF-α, HMGB-1, BUN, and creatinine concentrations, reducing mortality of LPS-induced AKI. These findings suggest that T-5224 may be protective against lethal LPS-induced AKI. T-5224 attenuated LPS-induced liver injury in mice. T-5224 may have beneficial effects in sepsis-induced organ dysfunctions. [3]
References:
[1]. Daisuke K. et al. Selective activator protein-1 inhibitor T-5224 prevents lymph node metastasis in an oral cancer model. Cancer Sci. 2016 May; 107 (5) 666–673.
[2]. Mari I. et al. T-5224, a selective inhibitor of c-Fos/activator protein-1, improves survival by inhibiting serum high mobility group box-1 in lethal lipopolysaccharide-induced acute kidney injury model. Journal of Intensive Care (2015) 3:49.
T-5224 是一种非肽类小分子 AP-1 抑制剂，特异性抑制 AP-1 与启动子区 AP-1 结合位点的结合，最初开发为抗炎药用于治疗类风湿性关节炎 (RA) 抑制炎症细胞因子和 MMPs 而没有任何副作用。[1]
体外研究表明，T-5224 对 MMP 表达的转录抑制不仅限于肿瘤细胞；它也发生在周围组织的 ECM 中，因此抑制肿瘤细胞浸润周围组织。此外，T-5224 通过转录抑制 MMP-2 和 -9 的表达，有效抑制转移、通过基底膜屏障浸润和迁移到 ECM 的基本步骤。[1]/p>\n
体内实验表明，T-5224 可阻断血清 TNF-α、HMGB-1、BUN 和肌酐浓度，降低 LPS 诱导的 AKI 的死亡率。这些发现表明 T-5224 可能对致命的 LPS 诱导的 AKI 具有保护作用。 T-5224 减轻 LPS 诱导的小鼠肝损伤。 T-5224 可能对败血症引起的器官功能障碍有益。 [3]
体内实验表明，T-5224 可阻断血清 TNF-α、HMGB-1、BUN 和肌酐浓度，降低 LPS 诱导的 AKI 的死亡率。这些发现表明 T-5224 可能对致命的 LPS 诱导的 AKI 具有保护作用。 T-5224 减轻 LPS 诱导的小鼠肝损伤。 T-5224 可能对败血症引起的器官功能障碍有益。 [3]
| Cas No. | 530141-72-1 | SDF | |
| 化学名 | 3-(5-(4-(cyclopentyloxy)-2-hydroxybenzoyl)-2-((3-oxo-2,3-dihydrobenzo[d]isoxazol-6-yl)methoxy)phenyl)propanoic acid | ||
| Canonical SMILES | O=C1NOC2=CC(COC(C=CC(C(C3=C(O)C=C(OC4CCCC4)C=C3)=O)=C5)=C5CCC(O)=O)=CC=C12 | ||
| 分子式 | C29H27NO8 | 分子量 | 517.53 |
| 溶解度 | ≥ 25.88mg/mL in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.9323 mL | 9.6613 mL | 19.3226 mL |
| 5 mM | 0.3865 mL | 1.9323 mL | 3.8645 mL |
| 10 mM | 0.1932 mL | 0.9661 mL | 1.9323 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
T-5224, a selective inhibitor of c-Fos/activator protein-1, improves survival by inhibiting serum high mobility group box-1 in lethal lipopolysaccharide-induced acute kidney injury model
Background: Sepsis is a potentially fatal syndrome mediated by an early [e.g., tumor necrosis factor-alpha (TNF-α)] and late [high mobility group box-1 (HMGB-1)] proinflammatory cytokine response to infection. Sepsis-induced acute kidney injury (AKI) is associated with a high mortality. C-Fos/activator protein-1 (AP-1) controls the transactivation of proinflammatory cytokines via AP-1 binding in the promoter region. T-5224 is a de novo small molecule inhibitor of c-Fos/AP-1 that controls gene expression of multiple proinflammatory cytokines. We investigated whether T-5224, a selective inhibitor of c-Fos/AP-1, improves survival in lethal lipopolysaccharide (LPS)-induced AKI by inhibiting early (TNF-α) and late (HMGB-1) proinflammatory cytokine response. Methods: Mice were divided into four groups (control, LPS, LPS + T-5224, and T-5224 only). Control mice were administered polyvinylpyrrolidone (PVP) solution orally, immediately after intraperitoneal (i.p.) saline injection. LPS mice were administered PVP solution orally immediately after i.p. LPS (10 mg/kg) injection. LPS + T-5224 mice were administered T-5224 orally (300 mg/kg) immediately after i.p. LPS injection. T-5224 mice were administered T-5224 orally (300 mg/kg) after i.p. saline injection. Serum concentrations of TNF-α, HMBG-1, and interleukin (IL)-10 were measured by enzyme-linked immunosorbent assay (ELISA). Serum blood urea nitrogen (BUN) and creatinine concentrations were commercially analyzed. Finally, histological examination was performed on the kidney. Results: Treatment with T-5224 decreased serum TNF-α and HMGB-1 levels and increased survival after LPS injection. Furthermore, T-5224 treatment decreased serum BUN and creatinine concentrations but increased serum IL-10 concentration. LPS-induced pathological changes in kidney were attenuated by T-5224 treatment. Conclusions: These results suggest that T-5224, a selective inhibitor of c-Fos/AP-1, inhibits expression of early and late proinflammatory cytokines, protecting mice from LPS-induced lethality. T-5224 is a potential approach for decreasing lethality in sepsis-induced AKI.
Single-cell transcriptomes reveal heterogeneity of high-grade serous ovarian carcinoma
Background: High-grade serous ovarian carcinoma (HGSOC) is the most common and aggressive histotype of epithelial ovarian cancer. The heterogeneity and molecular basis of this disease remain incompletely understood.
Methods: To address this question, we have performed a single-cell transcriptomics analysis of matched primary and metastatic HGSOC samples.
Results: A total of 13 571 cells are categorized into six distinct cell types, including epithelial cells, fibroblast cells, T cells, B cells, macrophages, and endothelial cells. A subset of aggressive epithelial cells with hyperproliferative and drug-resistant potentials is identified. Several new markers that are highly expressed in epithelial cells are characterized, and their roles in ovarian cancer cell growth and migration are further confirmed. Dysregulation of multiple signaling pathways, including the translational machinery, is associated with ovarian cancer metastasis through the trajectory analysis. Moreover, single-cell regulatory network inference and clustering (SCENIC) analysis reveals the gene regulatory networks and suggests the JUN signaling pathway as a potential therapeutic target for treatment of ovarian cancer, which is validated using the JUN/AP-1 inhibitor T-5224. Finally, our study depicts the epithelial-fibroblast cell communication atlas and identifies several important receptor-ligand complexes in ovarian cancer development.
Conclusions: This study uncovers new molecular features and the potential therapeutic target of HGSOC, which would advance the understanding and treatment of the disease.
OLFML2A is necessary for anti-triple negative breast cancer effect of selective activator protein-1 inhibitor T-5224
Previous studies have shown that expression of activator protein-1 (AP-1) family is significantly elevated in triple-negative breast cancer (TNBC), compared with that in other breast cancer subtypes. Here we investigated the anti-tumor effect and mechanism of T-5224, an inhibitor of c-Fos/AP-1, on TNBC. We identified that T-5224 inhibited the proliferation, migration, and invasion of TNBC cells and resulted in an increase in apoptosis. Furthermore, we found that OLFML2A is a key regulatory protein acting downstream of AP-1 and is involved in T-5224-targeted AP-1 action. Multiple clinical databases online have identified that high OLFML2A level is associated with poor prognosis in TNBC patients. In summary, our experimental and bioinformatic studies indicated that OLFML2A is necessary for AP-1-overexpressing TNBC. These findings demonstrate that AP-1-overexpressing TNBC dependent on OLFML2A, and targeting both AP-1 and OLFML2A through T-5224 may be a synergistic therapeutic strategy for this clinically challenging subset of breast cancer.
T-5224, a selective inhibitor of c-Fos/activator protein-1, attenuates lipopolysaccharide-induced liver injury in mice
The effect of T-5224, a selective inhibitor of c-Fos/activator protein (AP)-1, on lipopolysaccharide (LPS) induced liver injury was examined in mice. Administration of LPS (10 mg kg(-1), i.p.) markedly increased serum levels of tumor necrosis factor-alpha (TNFα), high mobility group box 1 (HMGB1), alanine aminotransferase/aspartate aminotransferase (ALT/AST), liver tissue levels of macrophage-inflammatory protein-1 alpha (MIP-1α) and monocyte chemoattractant protein-1 (MCP-1), as well as hepatic necrosis and inflammation, leading to 67 % lethality. Administration of T-5224 (300 mg kg(-1), p.o.) after intraperitoneal injection of LPS imparted appreciable protection against acute elevations in serum levels of TNFα, HMGB1, ALT/AST as well as in liver tissue levels of MIP-1α and MCP-1, and reduced the lethality (27 %). These data indicate that T-5224 ameliorates liver injury and improves survival through decreasing production of proinflammatory cytokines and chemokines in endotoxemic mice.
Selective activator protein-1 inhibitor T-5224 prevents lymph node metastasis in an oral cancer model
Activator protein-1 (AP-1) is a transcriptional factor that regulates the expression of various genes associated with tumor invasion and migration. The purpose of our study was to assess the therapeutic effects of a novel selective AP-1 inhibitor, T-5224, in preventing lymph node metastasis in head and neck squamous cell carcinoma (HNSCC) in an orthotopic mouse model. We assessed the effect of T-5224 on HNSCC cell invasion, migration, proliferation, and MMP activity by carrying out an in vitro study using an invasion assay, scratch assay, WST-8 assay, and gelatin zymography. We also observed morphological changes in HNSCC cells by time-lapse microscopy. Furthermore, cervical lymph node metastasis was assessed using an orthotopic tumor model of human oral squamous cell carcinoma cells (HSC-3-M3) injected in the tongue of a BALB/c nude mouse. T-5224 (150 mg/kg) or vehicle was given orally every day for 4 weeks. Animals were killed and assessed for lymph node metastasis by H&E staining of resected lymph nodes. T-5224 significantly inhibited the invasion, migration, and MMP activity of HNSCC cells in a dose-dependent manner; there was no significant influence on cell proliferation. The antimetastatic effect of T-5224 was also confirmed in our animal study. The rate of cervical lymph node metastasis in the model was 40.0% in the T-5224-treated group (n = 30) versus 74.1% in the vehicle-treated group (n = 27; P < 0.05). In conclusion, T-5224 inhibited the invasion and migration of HNSCC cells in vitro, and prevented lymph node metastasis in head and neck cancer in an animal model.