ABC99
目录号 : GC42669ABC99 是一种N-羟基海因（NHH）-氨基甲酸酯类、不可逆的Wnt去酰基化酶NOTUM选择性抑制剂。
Cas No.:2331255-53-7
Sample solution is provided at 25 µL, 10mM.
ABC99 is an N-hydroxyhydantoin (NHH)-carbamate-based, irreversible and selective inhibitor of Wnt-deacylating enzyme NOTUM[1]. NOTUM is a secreted serine hydrolase that negatively modulates Wnt/β-catenin signaling by removing the essential palmitoleate moiety from Wnt proteins[2]. ABC99 is usually used in models of osteoporosis, colorectal cancer and neurodegenerative diseases where Wnt signaling is insufficient[3][4].
In vitro, ABC99 (2µg/mL; 7–14 days) suppressed odontoblastic differentiation of human stem cells from apical papilla (hSCAPs), reduced ALP activity, blocked mineralized nodule formation, and decreased DSPP, DMP1 and ALP mRNA levels[5]. ABC99 (50nM and 500nM; 14 days) significantly reversed the inhibition of osteogenic differentiation of hPDLSCs by recombinant human NOTUM, restored ALP activity and calcium nodule formation, and increased the protein/mRNA levels of Osterix, OCN and Runx2[6].
In vivo, ABC99 (10mg/kg/day; p.o.; 4 weeks) almost completely prevented liver metastasis and lymph-node metastasis, reduced primary tumor weight, and significantly inhibited Ki67⁺ cell proliferation in orthotopic colorectal cancer mice[7].
References:
[1] Suciu RM, Cognetta AB 3rd, Potter ZE, Cravatt BF. Selective Irreversible Inhibitors of the Wnt-Deacylating Enzyme NOTUM Developed by Activity-Based Protein Profiling. ACS Med Chem Lett. 2018;9(6):563-568.
[2] Bayle ED, Svensson F, Atkinson BN, et al. Carboxylesterase Notum Is a Druggable Target to Modulate Wnt Signaling. J Med Chem. 2021;64(8):4289-4311.
[3] Zhao Y, Jolly S, Benvegnu S, Jones EY, Fish PV. Small-molecule inhibitors of carboxylesterase Notum. Future Med Chem. 2021;13(11):1001-1015.
[4] Larrick JW, Mendelsohn AR. Roads to the Fountain of Youth? Rejuvenating Intestinal Stem Cells. Rejuvenation Res. 2019;22(4):342-347.
[5] Zhao Q, Ren H, Wang N, Yuan X, Zhao Y, Wen Q. NOTUM plays a bidirectionally modulatory role in the odontoblastic differentiation of human stem cells from the apical papilla through the WNT/β-catenin signaling pathway. Arch Oral Biol. 2024;160:105896.
[6] Yang P, Li C, Kou Y, et al. Notum suppresses the osteogenic differentiation of periodontal ligament stem cells through the Wnt/Beta catenin signaling pathway. Arch Oral Biol. 2021;130:105211.
[7] Tian Y, Wang X, Cramer Z, et al. APC and P53 mutations synergise to create a therapeutic vulnerability to NOTUM inhibition in advanced colorectal cancer. Gut. 2023;72(12):2294-2306.
ABC99 是一种N-羟基海因（NHH）-氨基甲酸酯类、不可逆的Wnt去酰基化酶NOTUM选择性抑制剂[1]。NOTUM是一种分泌型丝氨酸水解酶，通过去除Wnt蛋白的必需棕榈酰基团来负向调控Wnt/β-catenin信号通路[2]。ABC99常用于骨质疏松、结直肠癌及神经退行性疾病等 Wnt 信号不足模型研究[3][4]。
在体外，ABC99（2µg/mL；7–14天）抑制人根尖乳头干细胞（hSCAPs）的成牙本质向分化，降低碱性磷酸酶（ALP）活性，阻断矿化结节形成，并降低牙本质涎磷蛋白（DSPP）、牙本质基质蛋白1（DMP1）和ALP的mRNA水平[5]。ABC99（50nM 和 500nM；14天）显著逆转重组人NOTUM对人牙周膜干细胞（hPDLSCs）成骨分化的抑制，恢复ALP活性和钙结节形成，并增加Osterix、骨桥蛋白（OCN）和骨钙素（Runx2）的蛋白/mRNA水平[6]。
在体内，ABC99（10mg/kg/天；口服；4周）几乎完全阻断了原位结直肠癌小鼠模型中的肝脏转移和淋巴结转移，降低了原发肿瘤的重量，并显著抑制了 Ki67⁺细胞增殖[7]。
| Cell experiment [1]: | |
Cell lines | Human stem cells from the apical papilla (hSCAPs) |
Preparation Method | Human stem cells from the apical papilla (hSCAPs) were recovered and cultured in a cell culture incubator under the conditions of 37°C with 5% CO2 (v/v). The culture medium was refreshed every 3 days. The normal proliferation medium (PM) contained α-MEM, 10% fetal bovine serum, and 1% penicillin. When cells reached 80% confluency, odontoblastic medium (OM) contained α-MEM, 10% fetal bovine serum, 50mg/L L-Ascorbic acid, 5mmol/L sodium β-phosphoglycerate, and 10nmol/L dexamethasone, was used to induce odontoblastic differentiation. To down-regulate the extracellular NOTUM level, ABC99, a small molecule inhibitor of NOTUM, was added to the OM medium at 2ug/ml. Cells were collected on odontoblastic induction day 3, 7, and 14. Alkaline phosphatase (ALP) staining, Alizarin red (ARS) staining and qRT-PCR were performed according to the manufacturer’s Instructions. |
Reaction Conditions | 2µg/mL; 7–14 days |
Applications | ABC99 suppressed odontoblastic differentiation of human stem cells from apical papilla (hSCAPs), reduced ALP activity, blocked mineralized nodule formation, and decreased DSPP, DMP1 and ALP mRNA levels. |
| Animal experiment [2]: | |
Animal models | C57BL/6J mice |
Preparation Method | 8-week-old, specific-pathogen-free C57BL/6J mice were kept in an air-conditioned room at a temperature of 23±1℃, a relative humidity of 50–70%, and a 12h light/dark cycle. APKS tumor organoids were orthostatic implanted into the colonic mucosa of C57BL/6J mice under endoscopic guidance to establish an in situ colorectal cancer model. Mice were treated with ABC99(10mg/kg/day; p.o.) or vehicle (corn oil) control after 4 weeks of engraftment and tumour growth. Mice are treated for indicated number of weeks (week 0-4 weeks post-implantation) (n=7 mice). Tumour weight, number of macrometastases in the liver and number of lymph node metastases in mice were qualified. Bioluminescent imaging (BLI) and Kaplan–Meier survival analysis were performed to monitor tumor burden and compare survival. |
Dosage form | 10mg/kg/day; p.o.; 4 weeks |
Applications | ABC99 almost completely prevented liver metastasis and lymph-node metastasis, reduced primary tumor weight in orthotopic colorectal cancer mice. |
References: | |
| Cas No. | 2331255-53-7 | SDF | |
| 化学名 | 7-(4-chlorobenzyl)-1,3-dioxohexahydroimidazo[1,5-a]pyrazin-2(3H)-yl-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate | ||
| Canonical SMILES | ClC1=CC=C(CN2CC(C(N(OC(N3C(C=CC=C4)=C4OCC3)=O)C5=O)=O)N5CC2)C=C1 | ||
| 分子式 | C22H21ClN4O5 | 分子量 | 456.9 |
| 溶解度 | 1mg/mL in Chloroform, 1mg/mL in acetonitrile, 1mg/mL in methyl acetate | 储存条件 | Store at -20°C |
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.1887 mL | 10.9433 mL | 21.8866 mL |
| 5 mM | 437.7 μL | 2.1887 mL | 4.3773 mL |
| 10 mM | 218.9 μL | 1.0943 mL | 2.1887 mL |
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