CZ415

Catalog No.S8298 Batch:S829801

Technical Data

Formula	C₂₂H₂₉N₅O₄S
Molecular Weight	459.56	CAS No.	1429639-50-8
Solubility (25°C)*	In vitro	DMSO	91 mg/mL (198.01 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

CZ415, a potent ATP-competitive mTOR inhibitor with very good cell permeability.

Targets

mTOR ^[1]
(Cell-free assay)

8.07(pIC50)

In vitro

CZ415 shows no genotoxic potential and has very good cell permeability. Treatment of CZ415 leads to inhibition of phosphorylation for downstream targets of mTORC1 and mTORC2(IC50=14.5 nM for pS6RP and IC50=14.8 nM for pAKT). The immunosuppressive effect of CZ415 is measured by detecting secreted IFNγ after 18 h in stimulated human whole blood, and the resulting IC50 was 226 nM. CZ415 shows no genotoxic potential. It is neither mutagenic in a bacterial mutation assay (Ames test) nor does it show genotoxicity in the mouse lymphoma assay (MLA), in either the presence or absence of rat-liver S9 mix^[1].

In vivo

In vivo studies show that CZ415 has moderate clearance and good oral bioavailability. In an anti-CD3 mouse model CZ415 efficiently inhibits mTOR downstream signaling and, in a CIA mouse model, shows significant antiinflammatory effects. With its extraordinary selectivity, drug-like properties and proven efficacy in vivo, CZ415 represents an ideal molecule for the pharmacological investigation of mTOR pathophysiological role in vivo^[1].

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines HEK294T17 cells Concentrations 3 µM start, 8 points 1:3 dilution steps. Incubation Time 2 h Method Cells are seeded in 90 µL DMEM containing 2% FCS at 4x10_⁴ cells/well (for pS6RP S240/244 assay) or 8x10_⁴ cells/well (for pAKT S473 assay) in a 96well U-bottom plate. The plate is then incubated for 1h in a humidified incubator (37ºC, 5% CO2) to allow cells to adhere. CZ415: 3 µM start, 8 points 1:3 dilution steps, n=2. Positive control: 1 μM PI-103 (n=8). Negative control: DMSO (n=8). 10 µL of 10x compound concentration in 1% DMSO/99% (DMEM 2% FCS) are added to the cells followed by 2 h incubation in a humidified incubator (37ºC, 5% CO2). Cells are lysed by addition of 10 μL 5x Complete Lysis Buffer and gentle shaking at 4°C for 15 min
Animal Study:^{^[1]}	Animal Models Male SD rats Dosages 1 mg/kg Administration i.v.

Cell Assay:^{^[1]}

Cell lines
HEK294T17 cells
Concentrations
3 µM start, 8 points 1:3 dilution steps.
Incubation Time
2 h
Method
Cells are seeded in 90 µL DMEM containing 2% FCS at 4x10_⁴ cells/well (for pS6RP S240/244 assay) or 8x10_⁴ cells/well (for pAKT S473 assay) in a 96well U-bottom plate. The plate is then incubated for 1h in a humidified incubator (37ºC, 5% CO2) to allow cells to adhere. CZ415: 3 µM start, 8 points 1:3 dilution steps, n=2. Positive control: 1 μM PI-103 (n=8). Negative control: DMSO (n=8). 10 µL of 10x compound concentration in 1% DMSO/99% (DMEM 2% FCS) are added to the cells followed by 2 h incubation in a humidified incubator (37ºC, 5% CO2). Cells are lysed by addition of 10 μL 5x Complete Lysis Buffer and gentle shaking at 4°C for 15 min

Animal Study:^{^[1]}

Animal Models
Male SD rats
Dosages
1 mg/kg
Administration
i.v.

References

[1] Cansfield AD, et al. ACS Med Chem Lett. 2016, 7(8):768-73.

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SHIPPING AND STORAGE
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