| 名稱 | KHS 101 |
| 描述 | KHS101 is a novel inhibitor of transforming acidic coiled-coil protein 3 (TACC3). It is a selective inducer of neuronal differentiation. |
| 細(xì)胞實(shí)驗(yàn) | Rat NPCs were derived and cultured as described previously by others. After hippocampal cell isolation, the number of dissociated cells was determined and ～5 × 10^5 cells were plated in 60-mm uncoated plates. After overnight incubation (37 °C, 5% CO2, and 95% humidity), the medium was changed and the cells were expanded and maintained in an undifferentiated state on polyornithine- (10 μg/mL in water) and laminin-coated (5 μg/mL in PBS;) dishes in DMEM/F12 supplemented with N2 and basic fibroblast growth factor (bFGF, 20 ng/mL;). For KHS101 and shRNA-induction experiments, early passage cells (passaged no more than six times after hippocampal isolation) were trypsinized and plated at a density of ～1,000 cells/cm2 into N2 medium (DMEM/F12 supplemented with N2) containing KHS analogs (e.g., KHS101, KHS92, and NP; SI Text) at different concentrations (0.5–5 μM) or DMSO (0.1%), RA (1–2 μM), BDNF (100 ng/mL), and/or BMP4 (50–100 ng/mL) for 4 d [1]. |
| 激酶實(shí)驗(yàn) | NPC lysate was prepared by sonication in PBS and protein samples were prepared at a concentration of 2 mg/mL. The benzophenone-KHS101 compound (KHS101-BP, 5 μM) was added to 50 μL of the proteome reaction with and without unlabeled compound (250 μM). Irradiation was for 1 h using a hand-held UV lamp at long wavelength (365 nm), and subsequently, a copper-catalyzed azide-alkyne cycloaddition reaction was performed. After incubation for 1 h at RT, proteins were precipitated using trichloroacetic acid and resuspended in isoelectric focusing sample buffer. 2D SDS/PAGE was performed using ReadyStripe IPG stripes following the manufacturer's protocol [1]. |
| 動(dòng)物實(shí)驗(yàn) | To investigate the pharmacokinetic properties of KHS101, male Sprague–Dawley rats were administered 3 mg/kg KHS101 i.v. or s.c. One rat was killed per time point at 5 min, 40 min, 1 h, and 3 h after dosing, and samples of blood (100 μL) and whole brains were collected. In a separate study, rats were administered 6 mg/kg KHS101 i.v. or s.c. Five blood samples of 100 μL each were collected serially via a jugular vein catheter at 2 min (i.v. only), 0.5 h (s.c. only), and 1, 3, 7 and 24 h after dosing. Plasma and homogenized whole brain samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). To study neuronal differentiation upon KHS101 administration in vivo, adult Fisher 344 rats (～10 wk old) received s.c. injections of 6 mg/kg KHS101 or vehicle control (5% ethanol in 15% Captisol). All rats received one daily i.p. injection of 200 mg/kg BrdU for 6 consecutive days after the first day. After 14 d, the animals were killed and perfusion fixed, and the brains were removed and subjected to immunohistochemical analysis [1]. |
| 體外活性 | KHS101能夠依賴劑量增加貼壁培養(yǎng)大鼠神經(jīng)前體細(xì)胞（NPCs）的神經(jīng)分化（EC50約1μM）。使用KHS101誘導(dǎo)的神經(jīng)元形成（在1.5-5μM KHS101條件下，40-60%的TuJ1+細(xì)胞）也在來(lái)源于成年大鼠海馬和側(cè)腦室（SVZ）的二代神經(jīng)球體形成條件下觀察到[2]。SMMC-7721和SK-Hep-1細(xì)胞在不同濃度的KHS101（分別為40μM和20μM）中培養(yǎng)，以確定IC50值。與對(duì)照（DMSO）處理相比，KHS101處理后球體的數(shù)量和大小顯著減少。球體形成依賴于KHS101的濃度。在KHS101存在下，Bmi1、c-Myc和Nanog的表達(dá)均降低。與對(duì)照（DMSO）相比，KHS101降低了p-AKT、p-GSK3β和β-catenin的表達(dá)，以及下游標(biāo)志物c-Myc和cyclin D1的表達(dá)[2]。 |
| 體內(nèi)活性 | 6 mg/kg KHS101（靜脈注射和皮下注射）的劑量產(chǎn)生了合理的血漿濃度（>1.5 μM），血漿半衰期為1.1-1.4小時(shí)，皮下給藥后的相對(duì)生物利用度為69%。最為重要的是，KHS101在大腦中的分布非常廣泛，這一點(diǎn)通過(guò)腦-血漿AUC(0-3h)比率約為8（給藥：3 mg/kg KHS101，靜脈注射）[1]得到了證明。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : 15 mg/mL
DMSO : 60 mg/mL (176.75 mM)
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| 關(guān)鍵字 | KHS-101 | KHS101 | KHS 101 |
| 相關(guān)產(chǎn)品 | 4-Isopropoxybenzoic acid | Docetaxel | Mebendazole | Flubendazole | Oxfendazole | Methylene Blue | Thiabendazole | Methylene Blue trihydrate | Griseofulvin | N-Phenylbenzylamine | Paclitaxel | 4'-Demethylepipodophyllotoxin |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | 細(xì)胞骨架化合物庫(kù) | NO PAINS 化合物庫(kù) | 干細(xì)胞分化化合物庫(kù) | 微管靶向化合物庫(kù) | 神經(jīng)元分化化合物庫(kù) |