HTH-01-015

HTH-01-015 is a potent and selective inhibitor of NUAK1 with a IC₅₀ value of 100nM and does not affect the activity of a panel of 139 other kinases, including additional AMPK family members^[1].

In vitro, HTH-01-015 (1–30μM) was employed for 3–5 days in MCF-10A and MDA-MD-231 PTEN⁺/PTEN^- isogenic breast cancer cell lines. PTEN^- cell lines were indeed more sensitive to HTH-01-015 treatment than PTEN⁺ cell lines, with significantly lower IC₅₀ values^[2]. HTH-01-015 (10µM) treated liver cancer cell lines SNU-387 and HepG2 for 48h and significantly reduced cell motility and invasiveness. HTH-01-015 treatment also up-regulated the epithelial marker E-cadherin while down-regulating mesenchymal markers N-cadherin and Vimentin, as well as matrix metalloproteinases MMP2 and MMP9^[3]. HTH-01-015 treatment (3μM) for 8h on U2OS osteosarcoma cells inhibited phosphorylation of the NUAK1 substrate MYPT1 and induced about 50% reduction in the population of cells in S-phase. HTH-01-015 treatment (3μM) for 10h also significantly inhibited mitotic entry^[4].

In vivo, HTH-01-015 (10 or 20mg/kg) was administered into CCl₄-induced liver fibrosis model mice by intraperitoneal injection every other day over 5–8 weeks. HTH-01-015 reduced fibrosis deposition and the expression of fibrosis-related proteins such as α-SMA and Collagen I in liver tissues^[5].

References:
[1] Banerjee S, Buhrlage S J, Huang H T, et al. Characterization of WZ4003 and HTH-01-015 as selective inhibitors of the LKB1-tumour-suppressor-activated NUAK kinases. Biochem J. 2014 Jan 1;457(1):215-25.Tang YC, Ho SC, Tan E, Ng AWT, McPherson JR, Goh GYL, Teh BT, Bard F, Rozen SG. Functional genomics identifies specific vulnerabilities in PTEN-deficient breast cancer. Breast Cancer Res. 2018 Mar 22;20(1):22.
[3] Chu D X, Jin Y, Wang B R, et al. LncRNA HOTAIR Enhances Epithelial-to-mesenchymal Transition to Promote the Migration and Invasion of Liver Cancer by Regulating NUAK1 via Epigenetic Inhibition miR-145-5p Expression. J Cancer. 2023 Jul 24;14(12):2329-2343.
[4] Banerjee S, Zagórska A, Deak M, Campbell D G, Prescott A R, Alessi D R. Interplay between Polo kinase, LKB1-activated NUAK1 kinase, PP1βMYPT1 phosphatase complex and the SCFβTrCP E3 ubiquitin ligase. Biochem J. 2014 Jul 15;461(2):233-45.
[5] You Y, Gao C Q, Wu J R, et al. Enhanced Expression of ARK5 in Hepatic Stellate Cell and Hepatocyte Synergistically Promote Liver Fibrosis. Int J Mol Sci. 2022 Oct 28;23(21):13084.

HTH-01-015是一种强效且选择性的NUAK1抑制剂，IC₅₀值为100nM，对包括其他AMPK家族成员在内的139种激酶均无活性影响^[1]。

体外实验中，HTH-01-015（1–30μM）处理MCF-10A及MDA-MB-231 PTEN⁺/PTEN^- 乳腺癌细胞系3–5天，PTEN^-细胞系对HTH-01-015的敏感性显著高于PTEN⁺细胞系，IC₅₀值明显降低^[2]。HTH-01-015（10μM）处理肝癌细胞系SNU-387和HepG2达48h，显著降低细胞运动与侵袭能力，同时上调上皮标志物E-cadherin，下调间质标志物N-cadherin和Vimentin，以及基质金属蛋白酶MMP2和MMP9^[3]。HTH-01-015（3μM）处理U2OS骨肉瘤细胞8h，可抑制NUAK1底物MYPT1的磷酸化，并使S期细胞比例减少约50%，同浓度处理10h还显著抑制细胞进入有丝分裂期^[4]。

体内实验中，将HTH-01-015（10或20mg/kg）经腹腔注射给予CCl₄诱导的肝纤维化模型小鼠，隔日一次，持续5–8周，HTH-01-015减少了肝组织纤维化沉积及α-SMA、Collagen I等纤维化相关蛋白的表达^[5]。