WWL0245 is a potent and specific BRD4 PROTAC. WWL0245 selectively degrades BRD4 with DC50 of <1 nM, and degrades BRD2/3 and PLK1 with DC50 of >1 μM). WWL0245 displays excellent selective cytotoxic effect in BETi-sensitive cancer cell lines such as AR-positive prostate cancer cell lines. WWL0245 is a promising drug candidate in AR-positive prostate cancer research and a valuable tool compound for studying the biological function of BRD4.

Physicochemical Properties

CAS #	2869057-11-2
Appearance	Typically exists as solid at room temperature
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)

Biological Activity

Targets	BRD4 1 nM (IC50)
ln Vitro	WWL0245 (0-1 μM; 96h) has an IC50 range of 0.0159 μM ~ 10 μM, which means that it inhibits the proliferation of AR-positive prostate cancer cells. Against AR-positive cell lines VCaP, LNCaP, and 22Rv1, it exhibited potent anti-proliferative activity at [1], with IC50 values of 0.016 μM, 0.021 μM, and 0.053 μM, in that order. The effects of WWL0245 (1 μM; 24 h) on LNCaP, 22Rv1, and VCaP cell lines included a concentration-dependent reduction in BRD4 levels and c-Myc levels as well as a time-dependent down-regulation of these proteins. Moreover, WWL0245 has no influence on c-Myc levels but can downregulate BRD4 levels in DU145 cells (1). The AR-regulated genes (PSA, TMPRSS2, ERG, FKBP5, BMPR1B) are all partially or completely inhibited by WWL0245 (100 nM-1 μM; 24 hours) [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: HL60, SU-DHL-6, RS4;11, JURKAT, A2780, MDA-MB- 468, BT549, LNCaP, 22Rv1, and VCaP cells Tested Concentrations: 0 μM -10 μM Incubation Duration: 96 h Experimental Results: Inhibited HL60, SU-DHL-6, RS4;11, JURKAT, A2780, MDA-MB-468, BT549, Western Blot Analysis[1] Cell Types: Prostate cancer Cell Types: LNCaP, 22Rv1, VCaP, DU145 Tested Concentrations: 1 nM, 10 nM, 100 nM, 1000 nM, 10000 nM Incubation Duration: 1 hour, 2 hrs (hours), 4 hrs (hours),12 hrs (hours), 24 hrs (hours),48 hrs (hours) Experimental Results: Resulted in the downregulation of BRD4 and c- Myc in a time/concentration-dependent manner in 22Rv1 and VCaP cells. RT-PCR[1] Cell Types: Prostate cancer: VCaP cell Tested Concentrations: 100 nM-1 μM Incubation Duration: 24 h Experimental Results: Suppressed the expression of AR-regulated genes in prostate cancer.
References	[1]. Rong Hu, et al. Identification of a selective BRD4 PROTAC with potent antiproliferative effects in AR-positive prostate cancer based on a dual BET/PLK1 inhibitor. Eur J Med Chem. 2022 Jan 5;227:113922.

Solubility Data

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)