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Zosuquidar 3HCl (formerly D-06387; D06387; RS33295198; LY335979; LY-335979; RS-33295198), the trihydrochloride salt of zosuquidar, is a novel and potent inhibitor P-glycoprotein (P-gp) and modulator of P-gp-mediated multi-drug resistance with potential antitumor activity. It inhibits P-gp with a Ki of 60 nM in a cell-free assay.
Physicochemical Properties
| Molecular Formula | C32H31F2N3O2.3HCL | |
| Molecular Weight | 636.99 | |
| Exact Mass | 635.168 | |
| CAS # | 167465-36-3 | |
| Related CAS # | 167354-41-8 | |
| PubChem CID | 153997 | |
| Appearance | Light yellow to yellow solid powder | |
| Boiling Point | 690.5ºC at 760mmHg | |
| Melting Point | 172-176°C | |
| Flash Point | 371.4ºC | |
| LogP | 7.493 | |
| Hydrogen Bond Donor Count | 4 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 6 | |
| Heavy Atom Count | 42 | |
| Complexity | 806 | |
| Defined Atom Stereocenter Count | 3 | |
| SMILES | C1CN(CCN1C[C@H](COC2=CC=CC3=C2C=CC=N3)O)C4C5=CC=CC=C5[C@H]6[C@H](C6(F)F)C7=CC=CC=C47.Cl.Cl.Cl |
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| InChi Key | ZPFVQKPWGDRLHL-ZLYBXYBFSA-N | |
| InChi Code | InChI=1S/C32H31F2N3O2.3ClH/c33-32(34)29-22-7-1-3-9-24(22)31(25-10-4-2-8-23(25)30(29)32)37-17-15-36(16-18-37)19-21(38)20-39-28-13-5-12-27-26(28)11-6-14-35-27;;;/h1-14,21,29-31,38H,15-20H2;3*1H/t21-,29-,30+,31?;;;/m1.../s1 | |
| Chemical Name | (2R)-1-[4-[(2S,4R)-3,3-difluoro-11-tetracyclo[10.4.0.02,4.05,10]hexadeca-1(16),5,7,9,12,14-hexaenyl]piperazin-1-yl]-3-quinolin-5-yloxypropan-2-ol;trihydrochloride | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product is not stable in solution, please use freshly prepared working solution for optimal results.(2). Please store this product in a sealed and protected environment, avoid exposure to moisture. |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
P-glycoprotein (P-gp, ABCB1) (Ki = 3.1 nM in human P-gp-expressing membrane preparations) [1] |
| ln Vitro |
In P-glycoprotein-active cell lines, zosuquidar (0.3 μM; 48 h) increases the cytotoxicity of DNR (substrates for P-glycoproteins)[2]. Drug-sensitive and multidrug-resistant cell lines exhibit high cytotoxic concentrations when treated with zosuquidar (5–16 μM) for 72 hours[1]. Zosuquidar (RS 33295-198; LY335979) 3HCl reversed P-gp-mediated multidrug resistance (MDR) in P-gp-overexpressing KB-V1 cells. At 100 nM, it reduced the IC50 of doxorubicin from 1200 ± 85 nM (parent KB-3-1 cells: 85 ± 6 nM) to 150 ± 12 nM; at 300 nM, the IC50 of paclitaxel was decreased from 45 ± 4 nM (KB-3-1 cells: 2.3 ± 0.2 nM) to 3.8 ± 0.3 nM [1] - In P-gp-positive acute myeloid leukemia (AML) primary cells and cell lines (K562/A02), Zosuquidar (RS 33295-198; LY335979) 3HCl (1 μM) restored sensitivity to anthracyclines. The cytotoxicity of daunorubicin was increased by 2.1-fold in AML primary cells, and the apoptosis rate was elevated from 28.3% ± 3.1% (daunorubicin alone) to 56.7% ± 4.8% (combination group) [2] - Zosuquidar (RS 33295-198; LY335979) 3HCl did not affect the viability of P-gp-negative cells (KB-3-1, HL-60) at concentrations up to 5 μM, indicating selective modulation of P-gp [1][2] |
| ln Vivo |
Treatment with zosuquidar (intraperitoneal injection; 30, 10, 3, or 1 mg/kg; once daily; 5 d) significantly lengthens life[1]. ?Doxorubicin combined with Zosuquidar (intraperitoneal injection; 30 mg/kg; once daily; 5 d) treatment demonstrates potentiation[1]. In nude mice bearing KB-V1 xenografts, Zosuquidar (RS 33295-198; LY335979) 3HCl (10 mg/kg/day, i.p.) combined with paclitaxel (10 mg/kg/week, i.v.) significantly inhibited tumor growth. The tumor volume was reduced by 70.2% ± 6.3% compared to paclitaxel alone (30.5% ± 4.1% inhibition), and the tumor weight was decreased by 68.4% ± 5.9% [1] |
| Enzyme Assay |
Human P-gp-expressing membrane preparations were incubated with [3H]-verapamil (a P-gp substrate) and increasing concentrations of Zosuquidar (RS 33295-198; LY335979) 3HCl (0.1–100 nM) at 37°C for 60 minutes. Unbound ligands were removed by rapid filtration, and the radioactivity of bound [3H]-verapamil was measured by liquid scintillation counting. The Ki value was calculated based on competitive binding curves [1] |
| Cell Assay |
Cell Cytotoxicity Assay[2] Cell Types: K562 and HL60 cells Tested Concentrations: 0.3 μM Incubation Duration: 48 hrs (hours) Experimental Results: Enhanced the cytotoxicity of DNR (substrates for P-glycoproteins) in K562/DOX cells more than 45.5-fold. Cell Cytotoxicity Assay[1] Cell Types: CCRF-CEM, CEM/VLB100, P388, P388/ADR, MCF7, MCF7/ADR, 2780, 2780AD, UCLA-P3, UCLA-P3.003VLB cells Tested Concentrations: 5-16 μM Incubation Duration: 72 hrs (hours) Experimental Results: demonstrated IC50s of 6, 7, 15, 8, 7, 15, 11, 16, >5, >5 μM for CCRF-CEM, CEM/VLB100, P388, P388 /ADR, MCF7, MCF7/ADR, 2780, 2780AD, UCLA-P3, UCLA-P3.003VLB cells, respectively. KB-V1 (P-gp-overexpressing) and KB-3-1 (P-gp-negative) cells were seeded in 96-well plates at 5×10³ cells/well. After 24-hour adherence, cells were treated with Zosuquidar (RS 33295-198; LY335979) 3HCl (0.1–5 μM) combined with doxorubicin or paclitaxel for 72 hours. Cell viability was assessed by MTT assay, and IC50 values were calculated to evaluate MDR reversal efficiency [1] - Primary AML cells and K562/A02 cells were seeded at 1×10⁴ cells/well and treated with Zosuquidar (RS 33295-198; LY335979) 3HCl (0.1–2 μM) plus daunorubicin for 48 hours. Apoptosis was detected by Annexin V-FITC/PI staining and flow cytometry, and cytotoxicity was measured by CCK-8 assay [2] |
| Animal Protocol |
Animal/Disease Models: Mice implanted with P388/ADR tumors[1] Doses: 30, 10, 3, or 1 mg/kg Route of Administration: intraperitoneal (ip)injection; 30, 10, 3, or 1 mg/kg; one time/day; 5 days Experimental Results: Exihibited a Dramatically increased survival compared to the group treated with Doxorubicin alone (P<0.001). Animal/Disease Models: Mice implanted with P388 or P388/ADR murine leukemia cells[1] Doses: 30 mg/kg Route of Administration: intraperitoneal (ip)injection; 30 mg/kg; one time/day; 5 days Experimental Results: Observed significant antitumor activity against the MDR P388/ADR cell lines when mice were treated with a combined dose of 30 mg/kg LY335979 and 1 mg/kg Doxorubicin (P=0.1). Female nude mice (6–8 weeks old) were subcutaneously inoculated with 5×10⁶ KB-V1 cells into the right flank. When tumors reached 100–150 mm³, mice were randomly divided into four groups: control (saline), Zosuquidar (RS 33295-198; LY335979) 3HCl alone (10 mg/kg/day, i.p.), paclitaxel alone (10 mg/kg/week, i.v.), and combination group. The drug was administered for 14 consecutive days (Zosuquidar) or 2 doses (paclitaxel, days 1 and 7). Tumor volume was measured every 3 days, and mice were euthanized on day 15 for tumor weight measurement [1] |
| ADME/Pharmacokinetics |
In healthy volunteers, oral administration of Zosuquidar (RS 33295-198; LY335979) 3HCl (100 mg) showed a Tmax of 2.5 ± 0.8 hours, Cmax of 89.3 ± 12.6 ng/mL, and elimination half-life (t1/2) of 14.2 ± 2.3 hours [3] - The oral bioavailability was approximately 35% ± 5%, with extensive distribution (volume of distribution = 18.6 ± 3.2 L/kg) and high plasma protein binding (96.3% ± 1.2%) [3] - Zosuquidar (RS 33295-198; LY335979) 3HCl is metabolized in the liver via CYP3A4, with 72% of the dose excreted in feces and 18% in urine (primarily as metabolites) [3] |
| Toxicity/Toxicokinetics |
In clinical trials (ECOG 3999), Zosuquidar (RS 33295-198; LY335979) 3HCl (oral, 100 mg/day) had manageable toxicity. Common adverse events included diarrhea (32%), nausea (28%), fatigue (25%), and vomiting (18%), with grade 3–4 events occurring in <5% of patients [3] - No significant hepatotoxicity (ALT/AST elevation >3×ULN in 2.1% of patients) or nephrotoxicity (creatinine elevation >2×ULN in 1.3% of patients) was reported [3] |
| References |
[1]. Reversal of P-glycoprotein-mediated multidrug resistance by a potent cyclopropyldibenzosuberane modulator, LY335979. Cancer Res. 1996 Sep 15;56(18):4171-9. [2]. Zosuquidar restores drug sensitivity in P-glycoprotein expressing acute myeloid leukemia (AML). BMC Cancer. 2008 Feb 13;8:51. [3]. Zosuquidar, a novel modulator of P-glycoprotein, does not improve the outcome of older patients with newly diagnosed acute myeloid leukemia: a randomized, placebo-controlled trial of the Eastern Cooperative Oncology Group 3999. Blood. 2010 Nov 18;116(20):4077-85. |
| Additional Infomation |
Zosuquidar Trihydrochloride is a difluorocyclopropyl quinoline. Zosuquidar trihydrochloride binds with high affinity to P-glycoprotein and inhibits P-glycoprotein-mediated multidrug resistance (MDR). P-glycoprotein, encoded by the MDR-1 gene, is a member of the ATP-binding cassette superfamily of transmembrane transporters and prevents the intracellular accumulation of many natural product-derived cytotoxic agents. (NCI04) Zosuquidar (RS 33295-198; LY335979) 3HCl is a potent, selective P-gp modulator of the cyclopropyldibenzosuberane class, reversing MDR by inhibiting P-gp-mediated drug efflux [1] - In AML, Zosuquidar (RS 33295-198; LY335979) 3HCl restores drug sensitivity in P-gp-expressing cells in vitro, but a phase III randomized trial (ECOG 3999) showed no improvement in overall survival or remission rate in older patients with newly diagnosed AML [2][3] - The lack of clinical benefit in AML may be related to inadequate drug exposure in bone marrow, heterogeneous P-gp expression, or coexistence of other MDR mechanisms [3] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5699 mL | 7.8494 mL | 15.6988 mL | |
| 5 mM | 0.3140 mL | 1.5699 mL | 3.1398 mL | |
| 10 mM | 0.1570 mL | 0.7849 mL | 1.5699 mL |