KP-457
目录号 : GC32452
KP-457 是一种选择性 ADAM17 抑制剂，具有反向异羟肟酸结构。
Cas No.:1365803-52-6
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
|
Cell lines |
TKDA3-4, a human iPSC clone; mouse C3H10T1/2 feeder cell line; control human platelets |
|
Preparation Method |
To generate CD41a+ MKs, human iPS-sac-derived HPCs were incubated with stem cell factor, thrombopoietin, and heparin on C3H10T1/2 feeder cells at 37°C during the MK differentiation phase (days14–20) and platelet production phase (days 20–24), followed by flow cytometric analysis. |
|
Reaction Conditions |
KP-457 (15μM), GM-6001 (50μM), or a p38 MAP kinase inhibitor (10μM) was administered during the MK differentiation and platelet production phases of HPCs cultivated at 37°C, followed by flow cytometric analysis. |
|
Applications |
KP-457 retains the expression of GPIbα on iPSC-Derived Platelets. KP-457 and BIRB796 exhibited an additive effect on the GPIbα+ platelet yield, which is consistent that they contributed to an increase in GPIbα+ iPSC platelets through separate mechanisms of action. |
|
References: [1]. Hirata S, et al. Selective Inhibition of ADAM17 Efficiently Mediates Glycoprotein Ibα Retention During Ex Vivo Generation of Human Induced Pluripotent Stem Cell-Derived Platelets. Stem Cells Transl Med. 2017 Mar;6(3):720-730. |
|
KP‐457 is a selective ADAM17 inhibitor, which has a reverse-hydroxamate structure. ADAM17, also known as TNF-α-converting enzyme, cleaves various molecules such as GPIbα, GPV, and TNF-α. KP‐457 inhibited cleavages of the TNF‐α sequence with 10 times the potency of GM‐6001 and was >50 times more selective for ADAM17 than for other MMPs and ADAM10 in cell‐free enzyme assays.[1]
The inhibition of C-terminal cleavage of the GPIbα sequence by ADAM17 was concentration dependent, with an IC50 of 10.6 nmol/l for KP-457. KP-457 at the lower concentration blocks Zn2+ chelation of the catalytic domain of ADAM17. Other studies confirmed that KP-457 exhibited neither genotoxicity nor systemic toxicity at doses up to 3 mg/kg administered to dogs intravenously once a day for 1 month. In vitro study also demonstrated that KP-457 could sustain intact GPIbα at levels seen in platelets freshly isolated from human blood. In addition, KP-457 inhibited GPIbα shedding with a potency 10 times that of GM-6001 in the cellular assay.[1]
References:
[1]. Hirata S, et al. Selective Inhibition of ADAM17 Efficiently Mediates Glycoprotein Ibα Retention During Ex Vivo Generation of Human Induced Pluripotent Stem Cell-Derived Platelets. Stem Cells Transl Med. 2017 Mar;6(3):720-730.
KP-457 是一种选择性 ADAM17 抑制剂，具有反向异羟肟酸结构。 ADAM17，也称为 TNF-α 转化酶，可裂解各种分子，例如 GPIbα、GPV 和 TNF-α。 KP-457 抑制 TNF-α 序列的裂解，效力是 GM-6001 的 10 倍，并且是 >；在无细胞酶测定中，对 ADAM17 的选择性比对其他 MMP 和 ADAM10 的选择性高 50 倍。[1]< /sup>
ADAM17 对 GPIbα 序列 C 末端裂解的抑制具有浓度依赖性，KP-457 的 IC50 为 10.6 nmol/l。较低浓度的 KP-457 阻断 ADAM17 催化结构域的 Zn2+ 螯合。其他研究证实，KP-457 在剂量高达 3 mg/kg 时每天一次静脉注射给狗，持续 1 个月，既没有表现出遗传毒性，也没有表现出全身毒性。体外研究还表明，KP-457 可以维持完整的 GPIbα，维持在从人体血液中新鲜分离的血小板中所见的水平。此外，在细胞试验中，KP-457 抑制 GPIbα 脱落的效力是 GM-6001 的 10 倍。[1]
| Cas No. | 1365803-52-6 | SDF | |
| Canonical SMILES | CS(=O)(NCC1=CC=C(C(N(C=O)O)CS(=O)(C2=CC=C(OCC#CC)C=C2)=O)C=C1)=O | ||
| 分子式 | C21H24N2O7S2 | 分子量 | 480.55 |
| 溶解度 | DMSO : 125 mg/mL (260.12 mM) | 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。 |
||
| 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 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.0809 mL | 10.4047 mL | 20.8095 mL |
| 5 mM | 0.4162 mL | 2.0809 mL | 4.1619 mL |
| 10 mM | 0.2081 mL | 1.0405 mL | 2.0809 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。
TNFR1 links TNF exocytosis to TNF production in allergen-activated RBL-2H3 cells
Cell Signal 2023 May;105:110607.PMID:36690134DOI:10.1016/j.cellsig.2023.110607.
We previously reported that the maximal production of Tumor Necrosis Factor (TNF or TNFα) in antigen-activated RBL-2H3 cells (a tumor analog of mucosal mast cells) requires Munc13-4, a regulator of exocytic fusion. In this study, we investigated the involvement of various fusion catalysts in TNF production. We observed a strong correlation between the total TNF level and TNF exocytosis in RBL-2H3 cells. RT-qPCR shows that TNFR1 (TNF receptor 1) is the sole TNFR expressed in these cells, and that its transcription is upregulated upon allergen-mediated activation. Importantly, the addition of soluble TNFR1 inhibits antigen-elicited TNF production in a dosage-dependent fashion. Likewise, TNF production is diminished in the presence of TACE (TNFα Converting Enzyme) inhibitor KP-457, which prevents the generation of soluble TNF (sTNF). Together, these findings indicate that sTNF and TNFR1 function as autocrine agent and receptor respectively at the mast cell surface to boost TNF proliferation during allergic inflammation.
Selective Inhibition of ADAM17 Efficiently Mediates Glycoprotein Ibα Retention During Ex Vivo Generation of Human Induced Pluripotent Stem Cell-Derived Platelets
Stem Cells Transl Med 2017 Mar;6(3):720-730.PMID:28297575DOI:10.5966/sctm.2016-0104.
Donor-independent platelet concentrates for transfusion can be produced in vitro from induced pluripotent stem cells (iPSCs). However, culture at 37°C induces ectodomain shedding on platelets of glycoprotein Ibα (GPIbα), the von Willebrand factor receptor critical for adhesive function and platelet lifetime in vivo, through temperature-dependent activation of a disintegrin and metalloproteinase 17 (ADAM17). The shedding can be suppressed by using inhibitors of panmetalloproteinases and possibly of the upstream regulator p38 mitogen-activated protein kinase (p38 MAPK), but residues of these inhibitors in the final platelet products may be accompanied by harmful risks that prevent clinical application. Here, we optimized the culture conditions for generating human iPSC-derived GPIbα+ platelets, focusing on culture temperature and additives, by comparing a new and safe selective ADAM17 inhibitor, KP-457, with previous inhibitors. Because cultivation at 24°C (at which conventional platelet concentrates are stored) markedly diminished the yield of platelets with high expression of platelet receptors, 37°C was requisite for normal platelet production from iPSCs. KP-457 blocked GPIbα shedding from iPSC platelets at a lower half-maximal inhibitory concentration than panmetalloproteinase inhibitor GM-6001, whereas p38 MAPK inhibitors did not. iPSC platelets generated in the presence of KP-457 exhibited improved GPIbα-dependent aggregation not inferior to human fresh platelets. A thrombus formation model using immunodeficient mice after platelet transfusion revealed that iPSC platelets generated with KP-457 exerted better hemostatic function in vivo. Our findings suggest that KP-457, unlike GM-6001 or p38 MAPK inhibitors, effectively enhances the production of functional human iPSC-derived platelets at 37°C, which is an important step toward their clinical application. Stem Cells Translational Medicine 2017;6:720-730.