生物活性

Selinexor(KPT-330), a first-in-class selective inhibitor of nuclear export (SINE)compound targeting exportin-1(XPO1), is currently in advanced phase II and IIIclinical trials against solid and hematologic cancers. Selinexor, analog of KPT-185, is an orally bioavailable selective CRM1 inhibitor. As the clinical candidate analog of KPT-185, it exhibits similar effects on the viability of T-ALL cells and elicits rapid apoptotic response. Selinexor reduces cell growth in MOLT-4, Jurkat, HBP-ALL, KOPTK-1, SKW-3, and DND-41 cell lines (IC50 = 34-203 nM).

Selinexorinhibits NF-κB activity with an IC50 of 6.72 μM.[1]

Anti-proliferative activity of selinexor

GIST,gastrointestinal stromal tumor; ASPS, alveolar soft part sarcoma

Cytotoxic effects of selinexor

体外研究

体内研究

2% DMSO+49% PEG 300+dd H2O

激酶实验

细胞实验

Cell culture and reagent[6]

Human liposarcoma cell line SW872, SA-4, LPS141,LP6, FU-DDLS-1, GOT3, MLS402, T7778 and T1000 were cultured and maintained in RPMI1640containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37^OCin a humidified atmosphere with 5% CO₂. Selinexor was dissolved indimethyl sulfoxide to a concentration of 20mM/L.

Cell proliferation assay (MTT assay)

Anti-proliferativeeffect of selinexor against liposarcoma cells was determined using colorimetricassays. Briefly, 4 × 10³ liposarcoma cells were seeded in quadrupletsin 96-well plates either in the presence or absence of selinexor at 37°C in aCO₂ incubator. At the conclusion of the experiment, 20μl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) was added to each well and incubated at 37°C in a CO₂ incubator for 2hr and then dissolved in 100μl of stop solution (SDS-HCl). Absorbancewas measured at 570 nm using a microplate reader. Doseresponse curves were plottedto calculate half-maximal inhibitory concentrations (IC50) for selinexor byGraphPad Prism4.

Colony formation assay

Liposarcomacells (1 × 10³) were seeded into 6-well plates in triplicates incomplete medium. After two days, media were supplemented with varyingconcentrations of selinexor. After 2 weeks, colonies were fixed with 5%glutaraldehyde and stained with crystal violet. For quantitative measurement,colonies were dissolved in 200μl of DMSO and absorbance was measured at570 nm using a microplate reader.

动物实验

Xenograft model[4]

Twenty nu/nu mice were inoculated subcutaneouslywith 2 × 10⁷ MDA-MB-231 cells. Treatment was initiated when thetumors reached a mean volume of 80mm³ (standard deviation ± 28.7mm³ ranges 50–157mm³). Mice were allocated to 2 groups of ten (10) micesuch that mean tumor volume in each group was within the range of 69 to 98 mm3.Mice were treated with vehicle or selinexor at 15mg/kg on a Monday, Wednesday andFriday (MWF) schedule. Body weight and condition of each animal were recordeddaily, and tumors were measured on MWF.

Immunohistochemistry

Formalin fixed paraffin embedded (FFPE)tissue blocks were sectioned at 4 microns, and deparaffinized through threechanges of xylenes and a decreasing series of ethanol. Antigen retrieval wasperformed in a steam cooker for 15 minutes in Declere working solution.Endogenous hydrogenase was blocked by incubation in 3% Hydrogen Peroxide forfive minutes. Slides were incubated in Casein-based protein block for 20minutes before incubation with ARRDC3 antibody at room temperature for 30minutes. Slides were then rinsed with buffer and incubated with Amplifier fromHi-Def Polymer Detection Kit for 10 minutes at room temperature. Afterwardsslides were rinsed with buffer and incubated in DAB chromogen for six minutesat room temperature for color development. The slides were counterstained withHematoxylin I, rinsed in water, and dehydrated through a series of increasingethanol and three changes of xylenes. Slides were then coverslipped. Digitalimages of slides were generated via Aperio AT scanner at 20×.Immunohistochemistry assays were performed on a Biogenex I6000 automatedstainer. The apoptag apoptosis assay and Masson’s Trichrome staining wereperformed manually per the manufacturer’s instructions.

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。

参考文献

[1] Kashyap T AC, Aboukameel A, Unger TJ, Klebanov B, Mohammad RM, Muqbil I, Azmi AS, Drolen C, Senapedis W, Lee M, Kauffman M, Shacham S, Landesman Y. Selinexor, a Selective Inhibitor of Nuclear Export (SINE) compound, acts through NF-κB deactivation and combines with proteasome inhibitors to synergistically induce tumor cell death. Oncotarget. 2016;7(48):78883-78895.
[2] Nakayama R ZY, Czaplinski JT, Anatone AJ, Sicinska ET, Fletcher JA, Demetri GD, Wagner AJ. Preclinical activity of selinexor, an inhibitor of XPO1, in sarcoma. Oncotarget. 2016 7(13):16581-16592.
[3] Gravina GL, Mancini A, Sanita P, et al. KPT-330, a potent and selective exportin-1 (XPO-1) inhibitor, shows antitumor effects modulating the expression of cyclin D1 and survivin [corrected] in prostate cancer models. BMC Cancer. 2015;15:941.
[more]

体内溶解度
5 mg/mL

Clinical Trial Information ( data from http://clinicaltrials.gov )

注：以上所有数据均来自公开文献，并不保证对所有实验均有效，数据仅供参考。

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