IITZ-01

IITZ-01 is a potent lysosomotropic autophagy inhibitor with single-agent antitumor activity, with an IC50 of 2.62 μM for PI3Kγ.

Product information

CAS Number: 1807988-47-1

Molecular Weight: 482.51

Formula: C26H23FN8O

Synonym:

Autophagy inhibitor IITZ-01

Chemical Name: N2-[4-(1H-1, 3-benzodiazol-2-yl)phenyl]-N4-(4-fluorophenyl)-6-(morpholin-4-yl)-1, 3, 5-triazine-2, 4-diamine

Smiles: FC1C=CC(=CC=1)NC1=NC(NC2C=CC(=CC=2)C2NC3=CC=CC=C3N=2)=NC(=N1)N1CCOCC1

InChiKey: XGEDDGLPNSLBFE-UHFFFAOYSA-N

InChi: InChI=1S/C26H23FN8O/c27-18-7-11-20(12-8-18)29-25-32-24(33-26(34-25)35-13-15-36-16-14-35)28-19-9-5-17(6-10-19)23-30-21-3-1-2-4-22(21)31-23/h1-12H,13-16H2,(H,30,31)(H2,28,29,32,33,34)

Technical Data

Appearance: Solid Power

Purity: ≥98% (or refer to the Certificate of Analysis)

Solubility: Soluble in DMSO

Shipping Condition: Shipped under ambient temperature as non-hazardous chemical or refer to Certificate of Analysis

Storage Condition: Dry, dark and -20 ^oC for 1 year or refer to the Certificate of Analysis.

Shelf Life: ≥360 days if stored properly.

Stock Solution Storage: 0 - 4 ^oC for 1 month or refer to the Certificate of Analysis.

Drug Formulation: To be determined

HS Tariff Code: 382200

How to use

In Vitro:

Sub-lethal concentrations of a lysosomotropic autophagy inhibitor, IITZ-01, sensitizes cancer cells (renal, lung, and breast carcinoma) to TRAIL-induced apoptosis through DR5 upregulation and survivin downregulation through ubiquitin-proteasome pathway. Knockdown of DR5 or overexpression of survivin inhibited combined treatment with IITZ-01 and TRAIL-induced apoptosis. IITZ-01 downregulated protein expression of Cbl, ubiquitin E3 ligase, and decreased expression level of Cbl markedly led to increase DR5 protein expression and TRAIL sensitivity. Moreover, IITZ-01 decreased expression level of survivin protein via downregulation of deubiquitinase ubiquitin-specific protease 9X (USP9X) expression. Taken together, these results provide the first evidence that IITZ-01 enhances TRAIL-mediated apoptosis through DR5 stabilization by downregulation of Cbl and USP9X-dependent survivin ubiquitination and degradation in renal carcinoma cells.

In Vivo:

The antitumor efficacy of IITZ-01 was investigated in in-vivo breast cancer model. MDA-MB-231/green fluorescent protein (GFP) orthotropic breast cancer xenografts were developed in CrTac:NCr-Foxnnu BALB/c female nude mice. Tumors after reaching ~100 mm3, vehicle, 45 mg/kg of IITZ-02, were administered through intraperitoneal (i.p.) route on every alternate day for 4 weeks. IITZ-01 reduced the active tumor burden around 8.8-fold when compared with control group as determined by tumor photon counts. Moreover, tumor volume measurements have demonstrated that IITZ-01 significantly inhibited average tumor growth when compared with control from third day of treatment (Fig. 7a–c). Significant reduction in average tumor weights was observed after treatment with test compounds compared with control (Fig. 7d). This treatment schedule of both compounds was well tolerated in mice for the total duration of administration with no significant changes in their body weights (Fig. 7e). In addition, hematoxylin and eosin staining revealed large areas of necrosis in treated groups of both compounds when compared with controls. Immunohistochemistry in histological sections revealed that exposure of both compounds have enhanced LC3-II- and PARP-positive staining in cells than vehicle control, indicating effective autophagy inhibition and triggering of apoptosis (Fig. 7f). Altogether, these findings demonstrated that IITZ-01 inhibits breast tumor growth by autophagy inhibition and apoptosis induction in vivo.

References:

1. Guntuku L, Gangasani JK, Thummuri D, Borkar RM, Manavathi B, Ragampeta S, Vaidya JR, Sistla R, Vegi NGM. IITZ-01, a novel potent lysosomotropic autophagy inhibitor, has single-agent antitumor efficacy in triple-negative breast cancer in vitro and in vivo. Oncogene. 2019 Jan;38(4):581-595. doi: 10.1038/s41388-018-0446-2. Epub 2018 Aug 30. PubMed PMID: 30166591.

Products are for research use only. Not for human use.