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| Targets |
SMARCA2 (BRM), SMARCA4 (BRG1), and PBRM1 (BAF180). AU-24118 is an orally bioavailable PROTAC degrader that targets these components of the mSWI/SNF chromatin remodeling complex.
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| ln Vitro |
In cell-based assays, AU-24118 degrades SMARCA2, SMARCA4, and PBRM1 with high potency and selectivity. The DC₅0 (concentration for 50% degradation) values are in the low nanomolar range. In VCaP prostate cancer cells, AU-24118 (100 nM, 24 hours) induces >90% degradation of SMARCA2, SMARCA4, and PBRM1. By degrading these core ATPases of the mSWI/SNF complex, AU-24118 disrupts the complex's function, leading to anti-proliferative and pro-apoptotic effects in cancer cells that depend on the mSWI/SNF complex for survival. AU-24118 demonstrates remarkable efficacy in in vitro models of prostate cancer, particularly in castration-resistant prostate cancer (CRPC).
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| Enzyme Assay |
A general cell-free protocol for assessing ternary complex formation: A TR-FRET-based assay is used to detect the formation of the ternary complex between the target protein (SMARCA2 or SMARCA4), the PROTAC (AU-24118), and the E3 ubiquitin ligase (e.g., VHL or CRBN). Recombinant His-tagged SMARCA2 (or SMARCA4) protein (10 nM) is incubated with recombinant GST-tagged VHL-elongin B-elongin C complex (50 nM) and varying concentrations of AU-24118 (0.01 nM to 10 uM) in assay buffer (25 mM HEPES, pH 7.5, 150 mM NaCl, 1 mM DTT, 0.01% Tween-20, 0.1% BSA). After 1 hour, europium-labeled anti-His antibody (2 nM) and streptavidin-d2 (20 nM) are added, along with an anti-GST antibody labeled with a donor fluorophore. The TR-FRET signal is measured after an additional hour. The EC₅0 for ternary complex formation is calculated. For target engagement, a cellular thermal shift assay (CETSA) can be used.
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| Cell Assay |
A general cellular protocol for assessing target degradation in VCaP cells: VCaP prostate cancer cells are seeded in 12-well plates at 2×10⁵ cells/well in DMEM containing 10% FBS. After 24 hours, the cells are treated with various concentrations of AU-24118 (0.1, 1, 5, 10, 25, 50, 100 nM) for 24 hours. For time-course studies, cells are treated with 10 nM AU-24118 for 0, 1, 2, 4, 6, 8, 12, 24 hours. Cells are harvested and lysed in RIPA buffer containing protease and phosphatase inhibitors. The lysates are centrifuged, and the supernatant is collected. Protein concentrations are quantified using the BCA assay. Equal amounts of protein (30-50 ug) are separated by SDS-PAGE and transferred to PVDF membranes. The membranes are probed with antibodies against SMARCA2, SMARCA4, PBRM1, and GAPDH (loading control). The DC₅0 and Dmax values are calculated by densitometry. For functional studies, cells are treated with AU-24118 for 72 hours, and cell viability is assessed using the MTT or CellTiter-Glo assay. Apoptosis is assessed by Annexin V/PI staining and flow cytometry, or by Western blot for cleaved PARP and cleaved caspase-3. Cell cycle analysis is performed by propidium iodide (PI) staining and flow cytometry.
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| Animal Protocol |
A general animal protocol for evaluating AU-24118 in a castration-resistant prostate cancer (CRPC) model: Male NSG mice are subcutaneously injected with 5×10⁶ VCaP cells. When tumors reach approximately 500-600 mm3, the mice are castrated (surgical castration). After castration, tumors regress and then regrow (castration-resistant growth). Once tumors regrow to approximately 200-300 mm3, the mice are randomized into treatment groups (n=10 per group). AU-24118 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administered via oral gavage at doses of 10, 30, and 100 mg/kg once daily for 28 days. Enzalutamide (standard of care AR antagonist) is administered at 10 mg/kg as a control or in combination with AU-24118. Tumor volume is measured twice weekly using a caliper, and body weight is recorded. Tumor growth inhibition (TGI) and tumor regression are calculated. At the end of the study, the mice are euthanized, and tumors are excised and weighed. Tumor tissues are processed for Western blot analysis to assess SMARCA2, SMARCA4, and PBRM1 degradation levels. Immunohistochemistry for Ki-67, cleaved caspase-3, and AR target genes (PSA, TMPRSS2) is performed.
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| ADME/Pharmacokinetics |
General pharmacokinetic protocol for AU-24118: Male Sprague-Dawley rats are administered AU-24118 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg) injection. The compound is formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. Plasma concentrations of AU-24118 are quantified by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated using non-compartmental analysis. AU-24118 is described as orally bioavailable.
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| Toxicity/Toxicokinetics |
General toxicity protocol for AU-24118: A 14-day repeated-dose oral toxicity study is performed in ICR mice. AU-24118 is administered via oral gavage at doses of 10, 30, and 100 mg/kg/day for 14 consecutive days. Clinical signs, body weight, and food consumption are monitored daily. At the end of the study, blood samples are collected for hematology (complete blood count with differential) and serum chemistry (ALT, AST, ALP, BUN, creatinine, total protein, albumin, glucose). Gross necropsy is performed, and the weights of major organs (liver, kidney, spleen, heart, lung, brain, testes) are recorded. Histopathological examination of these organs is conducted. Because the mSWI/SNF complex is involved in many cellular processes, the potential for on-target toxicity should be monitored, particularly in rapidly dividing tissues (bone marrow, gastrointestinal tract).
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| References | |
| Additional Infomation |
AU-24118 is a second-generation orally bioavailable PROTAC degrader of mSWI/SNF ATPases (SMARCA2/BRM and SMARCA4/BRG1) and PBRM1/BAF180. It is designed to degrade multiple components of the mSWI/SNF complex simultaneously. AU-24118 has demonstrated remarkable efficacy in in vitro and in vivo models, particularly in castration-resistant prostate cancer (CRPC). It induces tumor regression as a single agent and enhances the efficacy of enzalutamide, the standard of care AR antagonist in CRPC. AU-24118 is a research compound for studying the mSWI/SNF complex and for developing novel therapies for prostate cancer and potentially other malignancies driven by the mSWI/SNF complex.
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| Molecular Formula |
C37H40N6O4
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| Molecular Weight |
632.75
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| Appearance |
Light yellow to brown solid powder
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5804 mL | 7.9020 mL | 15.8040 mL | |
| 5 mM | 0.3161 mL | 1.5804 mL | 3.1608 mL | |
| 10 mM | 0.1580 mL | 0.7902 mL | 1.5804 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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