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GW9662

Alias: GW-9662; GW 9662; 2-Chloro-5-nitro-N-phenylbenzamide; 22978-25-2; 2-Chloro-5-nitrobenzanilide; GW-9662; benzamide, 2-chloro-5-nitro-N-phenyl-; 2-Chloro-5-nitro-N-4-phenylbenzamide; GW9662;
Cat No.:V0826 Purity: ≥98%
GW9662 (GW-9662; GW 9662) is an irreversible and selective inhibitor of PPAR (peroxisome proliferator-activated receptor) with potential anticancer activity.
GW9662
GW9662 Chemical Structure CAS No.: 22978-25-2
Product category: PPAR
This product is for research use only, not for human use. We do not sell to patients.
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Other Forms of GW9662:

  • GW9662-d5
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Top Publications Citing lnvivochem Products
Purity & Quality Control Documentation

Purity: ≥98%

Product Description

GW9662 (GW-9662; GW 9662) is an irreversible and selective inhibitor of PPAR (peroxisome proliferator-activated receptor) with potential anticancer activity. It inhibits PPARγ with an IC50 of 3.3 nM in a cell-free assay, and shows >100-fold selectivity for PPARγ over PPARα and PPARδ. GW9662 prevented activation of PPARgamma and inhibited growth of human mammary tumour cell lines. GW9662 may permit use of anti-ER strategies to inhibit breast cancer in ER- patients. GW9662 suppresses the cell viability with IC50 values ranging from 20-30μM.

Biological Activity I Assay Protocols (From Reference)
Targets
PPARγ: (IC50 = 3.3 nM)
PPARα: (IC50 = 32 nM)
PPARδ: (IC50 = 2000 nM)
ln Vitro
GW9662 has the ability to suppress radioligand binding to PPARγ, PPARα, and PPARδ, with corresponding pIC50 values of 8.48±0.27 (IC50=3.3 nM; n = 10), 7.49±0.17 (IC50=32 nM; n = 9), and 5.69±0.17 (IC50=2000 nM; n = 3). The binding studies with PPARα and PPARδ show that GW9662 is 10- and 600-fold less effective, respectively, with a nanomolar IC50 against PPARγ. GW9662 effectively and selectively blocks full-length PPARγ in cell-based reporter assays[1]. When paired with either 50 μM BRL 49653 (P=0.001) or 10 μM GW9662 (P=0.01) alone, co-treatment with both 50 μM BRL 49653 and 10 μM GW9662 leaves a significantly less number of viable cells after 7 days[2].
In the course of a high throughput screen to search for ligands of peroxisome proliferator activated receptor-gamma (PPARgamma), we identified GW9662 using a competition binding assay against the human ligand binding domain. GW9662 had nanomolar IC(50) versus PPARgamma and was 10- and 600-fold less potent in binding experiments using PPARalpha and PPARdelta, respectively. Pretreatment of all three PPARs with GW9662 resulted in the irreversible loss of ligand binding as assessed by scintillation proximity assay. Incubation of PPAR with GW9662 resulted in a change in the absorbance spectra of the receptors consistent with covalent modification. Mass spectrometric analysis of the PPARgamma ligand binding domain treated with GW9662 established Cys(285) as the site of covalent modification. This cysteine is conserved among all three PPARs. In cell-based reporter assays, GW9662 was a potent and selective antagonist of full-length PPARgamma. The functional activity of GW9662 as an antagonist of PPARgamma was confirmed in an assay of adipocyte differentiation. GW9662 showed essentially no effect on transcription when tested using both full-length PPARdelta and PPARalpha. Time-resolved fluorescence assays of ligand-modulated receptor heterodimerization, coactivator binding, and corepressor binding were consistent with the effects observed in the reporter gene assays. Control activators increased PPAR:RXR heterodimer formation and coactivator binding to both PPARgamma and PPARdelta. Corepressor binding was decreased. In the case of PPARalpha, GW9662 treatment did not significantly increase heterodimerization and coactivator binding or decrease corepressor binding. The experimental data indicate that GW9662 modification of each of the three PPARs results in different functional consequences. The selective and irreversible nature of GW9662 treatment, and the observation that activity is maintained in cell culture experiments, suggests that this compound may be a useful tool for elucidation of the role of PPARgamma in biological processes. [1]
This study demonstrated that the potent, irreversible and selective PPARgamma antagonist GW9662 prevented activation of PPARgamma and inhibited growth of human mammary tumour cell lines. Controversially, GW9662 prevented rosiglitazone-mediated PPARgamma activation, but enhanced rather than reversed rosiglitazone-induced growth inhibition. As such, these data support the existence of PPARgamma-independent pathways and question the central belief that PPARgamma ligands mediate their anticancer effects via activation of PPARgamma [2].
ln Vivo
Both BADGE- and GW9662(1 mg/kg, ip)-treated mice had significantly greater bone marrow (BM) nucleated cell counts than the aplastic anemia (AA) group[3]. In rats, GW9662 (1 mg/kg, ip) significantly reduces the renoprotective benefits of lipopolysaccharide (LPS)[4].
Researchers treated recipient mice with PPARγ antagonists BADGE or GW9662, or control vehicle. On day 14, mice were sacrificed and evaluated in PB by cell counts and in BM by estimating cellularity and morphologic examination of marrow adipocytes. Confocal microscopy showed massive expansion of adipocytes in the BM of AA mice. Overall BM cellularity, estimated by the density of 4′,6-diamidino-2-phenylindole (DAPI) staining (nuclear, blue color), was very low. In contrast, mice treated with PPARγ antagonists had many fewer adipocytes (green color, boron-dipyrromethene dye -BODIPY) in the BM and much higher marrow cellularity (Figure 1A, left). By conventional staining, the BM structure of AA mice showed extensive disruption and replacement by adipocytes, which occupied the “empty” space, whereas the BM of BADGE- or GW9662-treated mice showed less empty space and more hematopoietic cellularity (Figure 1A, right). Blood leukocyte and platelet counts were higher in BADGE-treated mice compared with AA mice (Figure 1B). BM nucleated cell counts in both BADGE- and GW9662-treated mice were significantly higher than counts in the AA group. Using flow cytometry, we found the frequency of Lin− cells and the absolute number of Lin−Sca1+c-kit+ (LSK) stem cells in BM were higher in BADGE-treated mice than in controls [3].
In our AA mice model, T cell immunity is the cause of BM destruction. In order to determine if treatment with PPARγ antagonists altered the immunological status of AA mice, we measured plasma cytokine levels in a multiplex assay. Insulin, an adipogenesis-related hormone, was higher in AA mice than in TBI controls; BADGE- or GW9662-treatment corrected insulin levels to normal. AA mice showed higher levels of the inflammatory cytokine monocyte chemoattractant protein-1 (MCP-1) and Th1 cytokines such as IFNγ, IFNγ-induced protein 10 (IP-10), and TNFα than did TBI control mice. BADGE- or GW9662-treatment reduced plasma MCP-1 as compared to levels in AA mice; BADGE reduced IFNγ and IP-10 levels, and GW9662-treatment reduced TNFα levels as well. BADGE- or GW9662-treatment tended to decrease IL-6 levels [3].
Effect of the GW9662 on renal/glomerular dysfunction caused by I/R in rats pretreated with LPS [4]
Rats that underwent renal I/R exhibited a significant increase in the serum level of creatinine compared with sham-operated ratsFigure 1a, suggesting a significant degree of renal dysfunction. Compared to rats subjected to I/R only (control), pretreatment of rats with LPS (1 mg/kg, IP) 24 hours prior to renal I/R significantly attenuated the increase in the serum level of creatinineFigure 1a. The attenuation in serum creatinine obtained after LPS pretreatment was reversed by administration of the specific PPARγ antagonist GW9662 (1 mg/kg, IP)Figure 1a.
In order to discount the possibility of a rapid increase in serum creatinine levels due to increased release of creatinine from muscle during I/R, creatinine clearance was also measuredFigure 1b. Rats subjected to renal I/R demonstrated a significant attenuation in creatinine clearance compared to sham-operated ratsFigure 1b, suggesting significant glomerular dysfunction. Compared to rats subjected to I/R only, administration of LPS 24 hours prior to I/R produced a modest, but significant, improvement in creatinine clearance. Most notably, this preservation of creatinine clearance afforded by LPS pretreatment was abolished by GW9662 Figure 1b.
Effect of GW9662 on tubular dysfunction caused by I/R in rats pretreated with LPS [4]
Renal I/R produced a significant increase in FENa, suggesting tubular dysfunctionFigure 2. Pretreatment of rats with LPS produced a significant attenuation in the I/R-mediated increase in FENa, suggesting improvement in tubular functionFigure 2. Administration of GW9662 in LPS-treated rats produced an increase in FENa, similar to that measured in control rats (I/R only), thus abolishing the protective effect mediated by LPSFigure 2. Renal I/R caused a nonsignificant decrease in urine flow compared with sham-operated ratsFigure 3. However, rats subjected to renal I/R that received LPS produced a significantly greater volume of urine compared to rats subjected to renal I/R aloneFigure 3. Administration of GW9662 to LPS-pretreated rats abolished this rise in urine flowFigure 3.
Effect of GW9662 on reperfusion injury caused by I/R in rats pretreated with LPS [4]
Renal I/R produced a significant increase in the serum concentrations of ASTFigure 4a and γ-GTFigure 4b in comparison with levels obtained from sham-operated rats. Serum concentrations of AST and γ-GT, which were used as markers of reperfusion injury, were significantly attenuated subsequent to administration of LPS (Figure 4a and B). Interestingly, GW9662 abolished the protective effect afforded by LPS pretreatment (Figure 4a and b).
Enzyme Assay
Binding Assays. [1]
SPAs for all three PPARs were performed as previously described for PPARγ. In brief, the human PPARα, PPARγ, and PPARδ ligand binding domains (LBDs) were expressed in E. coli as polyhistidine-tagged fusion proteins. Receptors were immobilized on SPA beads (Amersham Pharmacia) by addition of the desired receptor (15 nM) to a slurry of streptavidin-modifed SPA beads (0.5 mg/mL) in assay buffer. The mixture was allowed to equilibrate for at least 1 h at room temperature, and the beads were pelleted by centrifugation at 1000g. The supernate was discarded, and the beads were resuspended in the original volume of fresh assay buffer with gentle mixing. The centrifugation/resuspension procedure was repeated, and the resulting slurry of receptor-coated beads was used immediately or stored at 4 °C for up to 1 week before use. [3H]GW2331, [3H]rosiglitazone, and [3H]GW2443 were used as radioligands for determination of competition binding to PPARα, PPARγ, and PPARδ, respectively. Unless otherwise indicated, the buffer used for all assays was 50 mM HEPES (pH 7), 50 mM NaCl, 5 mM CHAPS, 0.1 mg/mL BSA, and 10 mM DTT. For some experiments, the HEPES (pH 7) was replaced with 50 mM Tris (pH 8).
Preparation of GW9662-Modified PPARγ for Mass Spectral Analysis. [1]
A stock solution of GW9662 in dimethyl sulfoxide was added to a 20 μM solution of PPARγ in 50 mM Tris (pH 8), 50 mM NaCl, 5 mM CHAPS, 0.1 mg/mL BSA, and 10 mM DTT. The final concentration of GW9662 was 40 μM. The solution was incubated at 4 °C followed by mass spectral analysis as described below.
Cell Assay
Cell-Based Reporter Assays.[1]
The ability of GW9662 to activate PPAR-mediated reporter gene transcription was assessed using GAL4 chimeras of the human receptors and a (UAS)5-tk-SPAP reporter plasmid as previously described for PPARγ, PPARα, and PPARδ. GW9662 antagonism of ligand-induced gene transcription was measured as previously described. Antagonism of agonist-induced reporter gene transcription was done by titrating varying concentrations of GW9662 in the presence of a constant concentration of activating ligand. The activating ligands used were 100 nM rosiglitazone for PPARγ, 8 nM GW7647 for PPARα, and 0.55 μM GW2433 for PPARδ, respectively.
The effects of GW9662 on activation of PPARγ, PPARα, and PPARδ were also assessed using full-length human receptors and a reporter construct, (L-FABP)4-tk-Dual-LUC, containing four copies of the L-FABP PPRE upstream of the minimal herpesvirus thymidine kinase promoter and a luciferase reporter gene. The receptor plasmids contained the appropriate full-length PPAR cDNA plus 9 base pairs of Kozak consensus sequence cloned into the TOPO site of pcDNA3.1-TOPO. Briefly, HEK293 cells were cultured in Minimum Essential Media containing 10% fetal calf serum, 1% penicillin/streptomycin, and 1% fungizone in a humidified incubator (5% CO2 in air) at 37 °C. The cells were seeded at 2 × 104 cells per well in 96 well culture plates the day prior to assay execution. Transfection was accomplished using PolyFect according to the manufacturers' instructions. Transfection mixtures for each well contained 0.167 μg of PPAR plasmid, 0.167 μg of LFABP reporter, and 0.167 μg of a renilla luciferase plasmid as transfection control. Cells were incubated with the transfection mixture for 5 h before treatment with either compound or vehicle for 48 h. Culture plates were assayed using the Dual Luciferase assay system according to the manufacturers' instructions.
MTT cell survival studies [2]
Rosiglitazone and GW9662 were used. Cells (MCF7, MDA-MB-231, MDA-MB-468) were plated in 96-well plates at a density of 1 × 103 cells per well in RPMI medium. After overnight incubation to allow for cell attachment, the medium was removed and replaced with fresh medium containing varying concentrations of rosiglitazone (1–100 μM), GW9662 (100 nM-50 μM) or solvent (dimethyl sulphoxide (DMSO)) alone. MDA-MB-231 cells were also subjected to combinations of rosiglitazone (10, 50 μM) and GW9662 (1, 10 μM) added simultaneously. The final concentration of DMSO in all cases did not exceed 0.1% and was not found to be cytotoxic in any of the cell lines tested at this concentration. Chemosensitivity was assessed following a continuous 72 h exposure using a standard 3-[4, 5-dimethylthiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay.
Cell growth assay [2]
MDA-MB-231 cells were seeded at a density of 1 × 105 cells per 25 cm3 tissue culture flask. After 24 h (day 0), the growth medium was replaced with fresh medium containing rosiglitazone (50 μM), GW9662 (10 μM) or both together. Control flasks received 0.1% DMSO. Cells were harvested on days 0, 3, 5, 7, 10 for each treatment condition by trypsinisation, stained using trypan blue, and the total and viable number of cells per flask calculated using a haemocytometer.
Preparation of nuclear extracts of treated cells for measurement of PPARγ and PPARα activity [2]
Levels of PPARγ and α activity following rosiglitazone (50 μM), GW9662 (10 μM) or combination treatment were measured in the nuclear fraction of MDA-MB-231 and MCF7 cells using the PPAR transfactor kit as described below. Cells were seeded at a density of 1 × 106 cells per 75 cm3 tissue culture flask. After 24 h, culture medium was removed and replaced with fresh medium containing the appropriate treatment. Following 2, 4, 8, 24 h of treatment, nuclear extracts were isolated (five flasks per treatment condition) using a transfactor extraction kit, as per the manufacturer's instructions.
Animal Protocol
Treatment of mice[3]
PPARγ antagonists, BADGE or GW9662, were dissolved in dimethyl sulfoxide (DMSO) and stored at −20°C. The aliquots were diluted with PBS to a final concentration of 10% DMSO and administrated by daily intraperitoneal injection at 30 mg/kg for BADGE, or at 1 mg/kg for GW9662, from one day prior to the experiment and continued for up to 2 weeks. In the FVB AA model, some mice were injected with cyclosporine A (CsA, 50 mg/kg/day) starting 1 hour after the LN injection, and continued for 5 days as immunosuppression. At the end of the experiments, the mice were euthanized by CO2 inhalation.
Methods, including peripheral blood (PB) and BM cell counting, flow cytometry, RNA isolation and gene expression analysis by PCR array, protein extraction and immunoblotting, cytokine measurement, histology using confocal microscopy, calcium flux assay and cell culture are detailed in the Online Supplementary Methods.
Experimental protocol[3]
Sixty-two male Wistar rats weighing 215 to 315 g were used in this study. Anesthetized rats were subjected to bilateral renal ischemia for 60 minutes followed by reperfusion for 6 hours. Animals were randomly allocated into 6 groups as follows: (1) I/R group: control, rats were administered 10% (v/v) DMSO (vehicle for GW9662, 1 mL /kg, IP) 24 and 12 hours prior to renal I/R, and saline (vehicle for LPS, 1 mL /kg, IP) 24 hours prior to renal I/R (N = 12); (2) I/R LPS group: rats were administered 10% (v/v) DMSO (vehicle for GW9662, 1 mL /kg, IP) 24 and 12 hours prior to renal I/R, and LPS (1 mg/kg, IP) 24 hours prior to renal I/R (N = 11); (3) I/R GW9662 group: rats were administered GW9662 (1 mg/kg, IP) 24 and 12 hours prior to renal I/R, and saline (vehicle for LPS, 1 mL /kg, IP) 24 hours prior to renal I/R (N = 9); (4) I/R LPS+GW9662 group: rats were administered GW9662 (1 mg/kg, IP) 24 and 12 hours prior to renal I/R, and LPS (1 mg/kg, IP) 24 hours prior to renal I/R (N = 11); (5) Sham group: rats were subjected to the same surgical procedures as above, except for renal I/R. Rats were administered 10% (v/v) DMSO (vehicle for GW9662, 1 mL /kg, IP) and saline (vehicle for LPS, 1 mL /kg, IP) at times equivalent to those described above (N = 12); (6) Sham GW9662 group: rats were subjected to the same surgical procedures as above, except for renal I/R. Rats were administered GW9662 (1 mg/kg, IP) and saline (vehicle for LPS, 1 mL /kg, IP) at times equivalent to those described above (N = 7).
The time and dose of LPS used were based on those previously shown by our group to provide protection against renal I/R injury in the rat. The time and dose of GW9662 used were chosen according to those previously shown to exert antagonism of PPARγ.
Dissolved in 10% (v/v) DMSO; 1 mg/kg; i.p. injection
Male Wistar rats
References

[1]. Functional consequences of cysteine modification in the ligand binding sites of peroxisome proliferator activated receptors by GW9662. Biochemistry. 2002 May 28;41(21):6640-50.

[2]. GW9662, a potent antagonist of PPARgamma, inhibits growth of breast tumor cells and promotes the anticancer effects of the PPARgamma agonist BRL 49653, independently of PPARgamma activation. Br J Pharmacol. 2004 Dec;143(8):933-7.

[3]. PPARγ antagonist attenuates mouse immune-mediated bone marrow failure by inhibition of T cell function.Haematologica. 2016 Jan;101(1):57-67.

[4]. The selective PPARgamma antagonist GW9662 reverses the protection of LPS in a model of renal ischemia-reperfusion. Kidney Int. 2005 Aug;68(2):529-36.

Additional Infomation
GW 9662 is a member of benzamides.
Peroxisome proliferator-activated receptor gamma (PPARgamma), a member of the nuclear receptor superfamily, is activated by several compounds, including the thiazolidinediones. In addition to being a therapeutic target for obesity, hypolipidaemia and diabetes, perturbation of PPARgamma signalling is now believed to be a strategy for treatment of several cancers, including breast. Although differential expression of PPARgamma is observed in tumours compared to normal tissues and PPARgamma agonists have been shown to inhibit tumour cell growth and survival, the interdependence of these observations is unclear. This study demonstrated that the potent, irreversible and selective PPARgamma antagonist GW9662 prevented activation of PPARgamma and inhibited growth of human mammary tumour cell lines. Controversially, GW9662 prevented rosiglitazone-mediated PPARgamma activation, but enhanced rather than reversed rosiglitazone-induced growth inhibition. As such, these data support the existence of PPARgamma-independent pathways and question the central belief that PPARgamma ligands mediate their anticancer effects via activation of PPARgamma.[2]
Acquired aplastic anemia is an immune-mediated disease, in which T cells target hematopoietic cells; at presentation, the bone marrow is replaced by fat. It was reported that bone marrow adipocytes were negative regulators of hematopoietic microenvironment. To examine the role of adipocytes in bone marrow failure, we investigated peroxisomal proliferator-activated receptor gamma, a key transcription factor in adipogenesis, utilizing an antagonist of this factor called bisphenol-A-diglycidyl-ether. While bisphenol-A-diglycidyl-ether inhibited adipogenesis as expected, it also suppressed T cell infiltration of bone marrow, reduced plasma inflammatory cytokines, decreased expression of multiple inflammasome genes, and ameliorated marrow failure. In vitro, bisphenol-A-diglycidyl-ether suppressed activation and proliferation, and reduced phospholipase C gamma 1 and nuclear factor of activated T-cells 1 expression, as well as inhibiting calcium flux in T cells. The in vivo effect of bisphenol-A-diglycidyl-ether on T cells was confirmed in a second immune-mediated bone marrow failure model, using different strains and non-major histocompatibility antigen mismatched: bisphenol-A-diglycidyl-ether ameliorated marrow failure by inhibition of T cell infiltration of bone marrow. Our data indicate that peroxisomal proliferator-activated receptor gamma antagonists may attenuate murine immune-mediated bone marrow failure, at least in part, by suppression of T cell activation, which might hold implications in the application of peroxisomal proliferator-activated receptor gamma antagonists in immune-mediated pathophysiologies, both in the laboratory and in the clinic. Genetically "fatless" mice developed bone marrow failure with accumulation of marrow adipocytes in our model, even in the absence of body fat, suggesting different mechanisms of systematic and marrow adipogenesis and physiologic versus pathophysiologic fat accumulation.[3]
Background: We have recently reported that pretreatment of rats with endotoxin (lipopolysaccharide, LPS) and selective agonists of the nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) protect the kidney against ischemia/reperfusion (I/R) injury. Here we investigate the hypothesis that the renoprotective effects of LPS may be due to an enhanced formation of endogenous ligands of PPARgamma, rather than an up-regulation of PPARgamma expression.

Methods: Rats were pretreated with LPS (1 mg/kg, IP, 24 hours prior to ischemia) in the absence (control) or presence of the selective PPARgamma antagonist GW9662 (1 mg/kg, IP, 24 and 12 hours prior to ischemia). Twenty-four hours after injection of LPS, rats were subjected to 60 minutes of bilateral renal ischemia, followed by 6 hours of reperfusion. Serum and urinary indicators of renal injury and dysfunction were measured, specifically serum creatinine, aspartate aminotransferase, and gamma-glutamyl-transferase, creatinine clearance, urine flow, and fractional excretion of sodium. Kidney PPARgamma1 mRNA levels were determined by reverse transcriptase-polymerase chain reaction.

Results: Pretreatment with LPS significantly attenuated all markers of renal injury and dysfunction caused by I/R. Most notably, GW9662 abolished the protective effects of LPS. Additionally, I/R caused an up-regulation of kidney PPARgamma1 mRNA levels compared to sham animals, which were unchanged in rats pretreated with LPS.

Conclusion: We document here for the first time that endogenous ligands of PPARgamma may contribute to the protection against renal I/R injury afforded by LPS pretreatment in the rat.[4]
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C13H9CLN2O3
Molecular Weight
276.67516207695
Exact Mass
276.03
Elemental Analysis
C, 56.43; H, 3.28; Cl, 12.81; N, 10.13; O, 17.35
CAS #
22978-25-2
Related CAS #
GW9662-d5;2117730-84-2
PubChem CID
644213
Appearance
White to off-white solid powder
Density
1.4±0.1 g/cm3
Boiling Point
360.9±32.0 °C at 760 mmHg
Melting Point
171-175 °C(lit.)
Flash Point
172.0±25.1 °C
Vapour Pressure
0.0±0.8 mmHg at 25°C
Index of Refraction
1.676
LogP
2.76
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
3
Rotatable Bond Count
2
Heavy Atom Count
19
Complexity
339
Defined Atom Stereocenter Count
0
SMILES
ClC1=CC=C(C=C1C(NC1C=CC=CC=1)=O)[N+](=O)[O-]
InChi Key
DNTSIBUQMRRYIU-UHFFFAOYSA-N
InChi Code
InChI=1S/C13H9ClN2O3/c14-12-7-6-10(16(18)19)8-11(12)13(17)15-9-4-2-1-3-5-9/h1-8H,(H,15,17)
Chemical Name
2-Chloro-5-nitro- N -phenylbenzamide
Synonyms
GW-9662; GW 9662; 2-Chloro-5-nitro-N-phenylbenzamide; 22978-25-2; 2-Chloro-5-nitrobenzanilide; GW-9662; benzamide, 2-chloro-5-nitro-N-phenyl-; 2-Chloro-5-nitro-N-4-phenylbenzamide; GW9662;
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

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO: 55 mg/mL (198.8 mM)
Water:<1 mg/mL
Ethanol:<1 mg/mL
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.04 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.5 mg/mL (9.04 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: ≥ 0.5 mg/mL (1.81 mM) (saturation unknown) in 1% DMSO 99% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.


Solubility in Formulation 4: 1% DMSO+30% polyethylene glycol+1% Tween 80: 30mg/mL

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Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 3.6143 mL 18.0714 mL 36.1428 mL
5 mM 0.7229 mL 3.6143 mL 7.2286 mL
10 mM 0.3614 mL 1.8071 mL 3.6143 mL

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(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • GW9662


    Large image of Figure 1.
    Effect of GW9662 on renal/glomerular dysfunction mediated by I/R in rats pretreated with lipopolysaccharide.Kidney Int.2005 Aug;68(2):529-36.
  • GW9662

    Large image of Figure 4.
    Effect of GW9662 on reperfusion injury mediated by I/R in rats pretreated with lipopolysaccharide.Kidney Int.2005 Aug;68(2):529-36.
  • GW9662

    Large image of Figure 2.
    Effect of GW9662 on tubular dysfunction subsequent to I/R in rats pretreated with lipopolysaccharide.

    Large image of Figure 3.

    GW9662

    Effect of GW9662 on urine flow subsequent to I/R in rats pretreated with lipopolysaccharide.Kidney Int.2005 Aug;68(2):529-36.

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