LY335979

• Catalog No.: S1481
• CAS Number: 167465-36-3
• M. F.: C32H34Cl3F2N3O2
• M. W.: 636.9870664
• Storage: -20°C

SYNONYMS

• IUPAC name: (2R)-1-{4-[(2R,4S,11S)-3,3-difluorotetracyclo[10.4.0.0^{2,4}.0^{5,10}]hexadeca-1(12),5(10),6,8,13,15-hexaen-11-yl]piperazin-1-yl}-3-(quinolin-5-yloxy)propan-2-ol trihydrochloride
• IUPAC Traditional name: (2R)-1-{4-[(2R,4S,11S)-3,3-difluorotetracyclo[10.4.0.0^{2,4}.0^{5,10}]hexadeca-1(12),5(10),6,8,13,15-hexaen-11-yl]piperazin-1-yl}-3-(quinolin-5-yloxy)propan-2-ol trihydrochloride
• Synonyms: Zosuquidar trihydrochloride; RS-33295-198

DATABASE IDS

• CAS Number: 167465-36-3

PROPERTIES

• Target: P-gp
• Salt Data: HCL
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Cancer

Biological Activity

• Description: LY335979 (Zosuquidar) is a potent modulator of P-glycoprotein-mediated multidrug resistance with K_i of 60 nM.
• Targets: P-glycoprotein
• IC50: 60 nM (K_i) ^[1]
• In Vitro: LY335979 competitively inhibits equilibrium binding of [^3H]vinblastine to Pgp by blocking [^3H]azidopine photoaffinity labeling of the Pgp in CEM/VLB₁₀₀ plasma membranes. ^[1] LY335979 alone shows the cytotoxicity to drug-sensitive and MDR cell lines with IC50 ranging from 6 μM-16 μM and produces its ability to completely reverse the resistance of the oncolytics (vinblastine, doxorubicin, or etoposide) to the MDR cell lines P388/ADR, MCF7/ADR, 2780AD, or UCLA-P3.003VLB at concentration of 0.1 and 0.5 μM. ^[1] LY335979 significantly restores drug sensitivity in P-gp-expressing leukemia cell lines including K562/HHT40, K562/HHT90, K562/DOX and HL60/DNR, and enhances the cytotoxicity of anthracyclines (daunorubicin, idarubicin, mitoxantrone) and gemtuzumab ozogamicin (Mylotarg) in primary AML blasts with active P-gp. ^[2] A latest paper indicates that LY335979 completely inhibits apically directed transport of (Z)-endoxifen in the ABCB1-transduced cells. ^[3]

Combination Therapy

• Description: In the MDR P388/ADR mouse model, combined treatment of LY335979 (30 mg/kg) and doxorubicin (1 mg/kg) exhibits a significant antitumor activity than doxorubicin alone. In UCLA-P3.003VLB human non-small cell lung carcinoma xenografts model, combined treatment of Taxol (20 mg/kg) and LY335979 (30 mg/kg) leads to a significant suppression of solid tumor growth compared to either treatment given alone. ^[1]

Protocol

• Protocol: Kinase Assay ^[1]
• ATPase Assay: P-Glycoprotein ATPase activity is measured by the liberation of inorganic phosphate from ATP. The assay is measured in a 96-well plate for 90 min at 37 °C. Membranes (8 μg-10 μg protein) are incubated in a total volume of 100 μL of buffer A containing 5 mM sodium azide, 1 mM ouabain, 1 mM EGTA, 3 mM ATP, an ATP regenerating system composed of 5 mM phosphoenolpyruvate, and 3.6 units/mL pyruvate kinase in the presence and absence of 1 mM sodium vanadate. Pgp-ATPase activity is defined as the vanadate-sensitive portion of the total ATPase activity. Plates are read 3 minutes after the addition of the detection solution. The absorbance is measured at 690 nm by a microtiter dish reader. A phosphate standard curve is used to calculate the μmol of phosphate formed. Samples are measured in triplicate.
• Protocol: Cell Assay ^[1]
• Cell Lines: CEM/VLB₁₀₀, P388/ADR, MCF7/ADR, 2780AD, and UCLA-P3.OO3VLB cells
• Concentrations: 0.05 μM to 5 μM
• Incubation Time: 72 hours
• Methods: Cell viability is determined using a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye reduction method. Cells are harvested during logarithmic growth phase, and seeded in 96-well plates. The cells are then cultured for 72 hours in the presence of oncolytics with or without modulators. MCF-7 and MCF-7/ADR cells are incubated 24 hours before the addition of the drug with and without the LY335979. LY335979 is prepared as 2 nM DMSO stocks and added to wells to give final concentrations ranging from 0.05 to 5 μM. After 72 hours, 20 μL of freshly prepared 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (5 mg/mL in Dulbecco's PBS) is added to each well and incubated for 4 hours in a 37 °C incubator containing 5% CO₂. Cells are pelleted in a Sorvall RT6000B centrifuge, 70 μL of medium is carefully removed from each well, and 100 μL of 2-propanol/0.04 N HC1 is added. Cells are resuspended 5-10 times with a Multipipettor or until no particulate matter is visible. Plates are immediately read on a Titertek Multiskan MCC/340 microplate reader Flow Laboratories with a test wavelength of 570 nm and a reference wavelength of 630 nm. Controls are measured in quadruplicate and modulators are measured in duplicate. Cytotoxicity analyses are also performed using the CeliTiter 96 AQueous assay kit.
• Protocol: Animal Study ^[1]
• Animal Models: P388 or P388/ADR cells are implanted by i.p. injection into female BDF₁ mice.
• Formulation: LY335979 is dissolved in 5% mannitol.
• Doses: ≤30 mg/kg
• Administration: Administered via i.p. and i.v.

References

• References: [1] Dantzig AH, et al. Cancer Res. 1996, 56(18), 4171-4179.
• References: [2] Tang R, et al. BMC Cancer. 2008, 8,51.
• References: [3] Iusuf D, J Pharmacol Exp Ther. 2011, 337(3), 710-717.

REFERENCES

• Dantzig AH, et al. Cancer Res. 1996, 56(18), 4171-4179.
• Tang R, et al. BMC Cancer. 2008, 8,51.
• Iusuf D, J Pharmacol Exp Ther. 2011, 337(3), 710-717.