| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg | |||
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| Targets |
Two primary targets: NGLY1 (N-glycanase 1; PNGase) and Nrf1 (Nuclear respiratory factor 1). WRR139 inhibits NGLY1 activity, which in turn disrupts the NGLY1-mediated deglycosylation, maturation, and nuclear localization of the transcription factor Nrf1. This leads to the suppression of Nrf1's target genes, which are primarily genes encoding proteasome subunits and other components of the ubiquitin-proteasome system.
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| ln Vitro |
WRR139 (0.01-100 μM; 1 hour) suppresses Nrf1 processing in HEK293 cells and inhibits NGLY1 in vitro [1]. In HEK293 cells, Nrf1 processing, localization, and activation are disrupted by WRR139 (5 μM; 18 hours) [1]. In U266, H929, and Jurkat cells, WRR139 (1 μM; 24 hours) increases carfilzomib cytotoxicity in an NGLY1-dependent manner [1].
In a cell-free enzymatic assay, WRR139 inhibits NGLY1 (rhNGLY1) with an IC50 of <10 microM, using S-alkylated RNase B as the substrate. In cell-based assays, it impairs the processing and activation of Nrf1 in HEK293 cells at concentrations as low as 0.01 microM. This leads to a decrease in the compensatory expression of proteasome genes. It does not inhibit caspases at concentrations <10 microM, indicating selectivity. |
| ln Vivo |
In a multiple myeloma xenograft model, treatment with WRR139 (1 microM) significantly potentiated the cytotoxicity of the proteasome inhibitor carfilzomib (1-100 nM). This effect was observed in multiple myeloma (U266 & H929), T-ALL (Jurkat), and HeLa cells, but was absent in NGLY1-knockdown cells, confirming the on-target mechanism. This demonstrates that blocking Nrf1 activation can overcome resistance to proteasome inhibitors.
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| Enzyme Assay |
An in vitro enzymatic assay for NGLY1 activity is performed by incubating recombinant NGLY1 protein with a standard substrate, such as S-alkylated RNase B (a glycoprotein). The reaction proceeds in the presence or absence of varying concentrations of WRR139. The de-glycosylation of the substrate is monitored, typically by gel electrophoresis (a shift in molecular weight) or by using a fluorescently labeled substrate, to calculate the IC50.
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| Cell Assay |
To assess cellular NGLY1 inhibition, a reporter cell line can be used, such as ddVenus reporter K562 cells. These cells contain a reporter construct that is dependent on Nrf1 activation. Co-treatment with a low dose of proteasome inhibitor (e.g., 1 microM carfilzomib) causes proteotoxic stress, inducing the Nrf1 response and activating the reporter. Treatment with WRR139 blocks this activation, which is quantified by measuring fluorescence or luminescence, yielding an IC50 of about 5.5 microM.
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| Animal Protocol |
The in vivo activity of WRR139 is primarily studied in xenograft mouse models. Immunodeficient mice are implanted subcutaneously with human cancer cells, such as multiple myeloma (U266) or T-ALL (Jurkat) cells. After tumors are established, mice are treated with vehicle, carfilzomib alone, WRR139 alone, or the combination. Tumor volumes are measured regularly. Potentiation of carfilzomib's efficacy is assessed by comparing the combination group to the carfilzomib monotherapy group.
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| ADME/Pharmacokinetics |
For in vivo studies, WRR139 is formulated as a powder and dissolved in DMSO to a stock solution. It is then diluted in appropriate vehicles, such as 5% DMSO, 40% PEG300, 5% Tween 80, and 50% ddH2O, to achieve the desired concentration for injection. The compound is typically administered intraperitoneally (IP). Its solubility is low in aqueous solutions, requiring these formulation vehicles.
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| Toxicity/Toxicokinetics |
Specific toxicology data for WRR139 is not publicly available. However, a key aspect of its safety profile is its selectivity. It inhibits NGLY1 and Nrf1 while showing minimal off-target activity against caspases at concentrations used for NGLY1 inhibition (recommended <10 microM). It has been used in multiple mouse xenograft studies without reports of overt toxicity. Long-term toxicity studies would be required to assess its safety for therapeutic use.
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| References |
[1]. Tomlin FM, et al. Inhibition of NGLY1 Inactivates the Transcription Factor Nrf1 and Potentiates Proteasome Inhibitor Cytotoxicity. ACS Cent Sci. 2017 Nov 22;3(11):1143-1155.
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| Additional Infomation |
WRR139 is a research tool compound used to study the NGLY1-Nrf1 axis and its role in maintaining proteostasis, particularly in the context of cancer and resistance to proteasome inhibitors. Proteasome inhibitors (e.g., bortezomib, carfilzomib) are effective drugs for multiple myeloma, but resistance often develops via Nrf1-mediated upregulation of proteasome subunits. WRR139 blocks this adaptive response. The drug is not approved for clinical use, but it is a valuable probe for overcoming drug resistance in cancer.
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| Molecular Formula |
C25H32N2O4S
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|---|---|
| Molecular Weight |
456.5976
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| Exact Mass |
456.208
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| CAS # |
2138924-36-2
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| PubChem CID |
135300204
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| Appearance |
White to off-white solid powder
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| LogP |
4.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
11
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| Heavy Atom Count |
32
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| Complexity |
713
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC1=CC=C(C=C1)S(=O)(=O)/C=C/C(=O)N[C@@H](CCC2=CC=CC=C2)C(=O)NCCC(C)C
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| InChi Key |
BLEQGDOEDLJWCG-NGIFANIDSA-N
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| InChi Code |
InChI=1S/C25H32N2O4S/c1-19(2)15-17-26-25(29)23(14-11-21-7-5-4-6-8-21)27-24(28)16-18-32(30,31)22-12-9-20(3)10-13-22/h4-10,12-13,16,18-19,23H,11,14-15,17H2,1-3H3,(H,26,29)(H,27,28)/b18-16+/t23-/m0/s1
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| Chemical Name |
(2S)-N-(3-methylbutyl)-2-[[(E)-3-(4-methylphenyl)sulfonylprop-2-enoyl]amino]-4-phenylbutanamide
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO: 100 mg/mL (219.01 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.48 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1901 mL | 10.9505 mL | 21.9010 mL | |
| 5 mM | 0.4380 mL | 2.1901 mL | 4.3802 mL | |
| 10 mM | 0.2190 mL | 1.0951 mL | 2.1901 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(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.
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