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| Targets |
DCAF1 binder 1 specifically targets DCAF1 (also known as VprBP), a substrate receptor of the CUL4-DDB1 (CRL4) E3 ubiquitin ligase complex. The CRL4-DCAF1 complex is involved in the ubiquitination and proteasomal degradation of various cellular proteins. DCAF1 binder 1 acts as a ligand that binds to DCAF1 with high selectivity, occupying the substrate-binding pocket. By binding to DCAF1, the compound can be used as a “bait” to recruit the CRL4 ubiquitination machinery to a target protein of interest. This is typically achieved by conjugating DCAF1 binder 1 (via a linker) to a molecule that binds the target protein, forming a PROTAC. The PROTAC brings the target protein into proximity with DCAF1, leading to ubiquitination and degradation of the target. Thus, DCAF1 binder 1 enables the development of DCAF1-recruiting PROTACs.
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| ln Vitro |
In vitro, DCAF1 binder 1 binds selectively to the CRL4-DCAF1 E3 ligase complex, as demonstrated by surface plasmon resonance (SPR) or biolayer interferometry (BLI) experiments using recombinant DCAF1 protein or CRL4-DCAF1 complex. The binding affinity (KD) is typically in the nanomolar to low micromolar range, though the exact value is not specified. The compound itself does not induce degradation; it is a ligand that is intended to be used as a building block for PROTAC molecules. As a PROTAC component, DCAF1 binder 1 facilitates targeted protein degradation when conjugated to a target-binding warhead. The compound has no significant intrinsic biological activity beyond its DCAF1 binding, making it an ideal scaffold for recruiting this E3 ligase.
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| ln Vivo |
In vivo activity of DCAF1 binder 1 itself has not been reported because it is a research tool used primarily as a chemical building block for PROTACs rather than as a therapeutic entity. The in vivo activity of PROTACs that incorporate DCAF1 binder 1 has been demonstrated in preclinical models. For example, DCAF1-recruiting PROTACs against specific oncoproteins have been shown to degrade their targets and reduce tumor growth in xenograft mouse models. However, the parent ligand DCAF1 binder 1, when administered alone, is not expected to have any in vivo efficacy or measurable pharmacodynamic effect because it does not engage a specific disease-relevant target. It serves only as an E3 ligase recruiter.
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| Enzyme Assay |
DCAF1 binder 1 is used in biochemical binding assays to measure its affinity for the DCAF1 protein or the CRL4-DCAF1 complex. The protocol for Surface Plasmon Resonance (SPR) analysis is as follows: Recombinant human DCAF1 protein (full-length or the WD40 domain) is immobilized onto a CM5 sensor chip via amine coupling. The running buffer is HBS-EP (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% Tween-20). DCAF1 binder 1 is dissolved in DMSO and serially diluted in running buffer (0.1 nM - 10 uM, final DMSO 1%). The compound solutions are injected over the DCAF1 surface at a flow rate of 30 uL/min for 60 s (association), followed by a 180 s dissociation phase. The chip surface is regenerated with 10 mM glycine-HCl (pH 2.0). Sensorgrams are double-referenced and globally fit to a 1:1 Langmuir binding model to calculate ka, kd, and KD. The KD for DCAF1 binder 1 has been reported to be in the low nanomolar range (specific value not available). Alternatively, a TR-FRET (time-resolved fluorescence energy transfer) binding assay using labeled DCAF1 and a fluorescent tracer can be developed. DCAF1 binder 1 is also used in competition binding assays to determine the IC50 for displacement of a known DCAF1 ligand.
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| Cell Assay |
Cellular assays to validate DCAF1 binder 1-based PROTACs are performed in cultured cancer cell lines (e.g., HEK-293T, HeLa, MCF-7, or specific cancer lines dependent on the target protein). DCAF1 binder 1 alone is used as a negative control to demonstrate that binding to DCAF1 is not sufficient to induce degradation. The standard protocol for a PROTAC compound (composed of DCAF1 binder 1 + linker + target binder) involves treating cells with the PROTAC at various concentrations (e.g., 0.1 nM - 10 uM) for 4-24 h. Cells are lysed, and the abundance of the target protein of interest (e.g., an oncoprotein) is assessed by Western blot or by quantitative immunoassay. Degradation is quantified by comparing target protein levels relative to a loading control (e.g., GAPDH, beta-actin). DCAF1 binder 1 (alone, at 1-10 uM) is used as a control to confirm that degradation depends on the bivalent PROTAC structure. Additionally, a competitive experiment with excess DCAF1 binder 1 pre-incubated with cells can block PROTAC-induced degradation, confirming that the effect is DCAF1-dependent. The DC50 (concentration for 50% degradation) and Dmax (maximal degradation) are calculated. The ubiquitination of the target protein can be confirmed by immunoprecipitation of the target followed by Western blot for ubiquitin. Because DCAF1 binder 1 itself is not a therapeutic agent, no specific cell-based assay for the ligand alone (without linker and target warhead) is performed beyond binding and competition experiments.
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| Animal Protocol |
In vivo efficacy studies are not performed with DCAF1 binder 1 alone, as it has no therapeutic effect. For PROTACs incorporating DCAF1 binder 1, mouse xenograft studies are standard. A typical protocol: Female NOD/SCID or NSG mice are injected subcutaneously with cancer cells (e.g., 5 × 10⁶ cells) expressing the target protein. When tumors reach 150-200 mm3, mice are randomized (n = 8-10/group). The DCAF1-recruiting PROTAC is formulated in 10% DMSO/40% PEG300/5% Tween-80/45% saline or in 20% Captisol in PBS. The PROTAC is administered intraperitoneally or intravenously at doses of 1-30 mg/kg, daily or every other day, for 2-4 weeks. Tumor volume is measured with calipers twice weekly. Body weight is monitored for toxicity. The vehicle group receives the same formulation without the PROTAC. A control group receives a PROTAC with a mutated (inactive) DCAF1 binder 1 to confirm on-target degradation is required for efficacy. At study termination, tumors are excised, weighed, and processed for Western blot analysis to confirm target degradation, for immunohistochemistry (Ki67, cleaved caspase-3), and for histology. These experiments demonstrate that DCAF1-recruiting PROTACs can achieve significant tumor growth inhibition. For the free DCAF1 binder 1 ligand, no such studies are performed because it lacks a target-binding warhead.
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| ADME/Pharmacokinetics |
Pharmacokinetic studies are not typically performed with DCAF1 binder 1 itself because it is a research chemical intermediate rather than a drug candidate. However, the physicochemical properties of DCAF1 binder 1 can be predicted. It has a molecular weight of 507.07 g/mol. The compound is likely to have moderate lipophilicity and may have acceptable permeability. The ligand is intended for use as a building block for PROTACs. In the context of a PROTAC molecule (full degrader), the PK properties will be dominated by the overall PROTAC (which typically has a higher molecular weight, >1000 Da) rather than the DCAF1-binding ligand alone. DCAF1 binder 1 is soluble in DMSO and in DMSO/PEG/Tween/saline mixtures for in vivo formulation when conjugated into a PROTAC. No detailed PK parameters (t½, Cmax, AUC, clearance, protein binding) for DCAF1 binder 1 have been published. The compound is for research use only.
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| Toxicity/Toxicokinetics |
The toxicity of DCAF1 binder 1 has not been formally studied because the compound is used as a research tool for targeted protein degradation. It is not a drug candidate. The predicted toxicity is expected to be low, as DCAF1 is not an essential gene in most adult tissues, and DCAF1-binding ligands are generally well-tolerated in preclinical models when used as part of a PROTAC. No specific toxicity data (LD50, acute toxicity, genotoxicity, organ toxicity) are available. However, as with any small-molecule research chemical, standard laboratory safety precautions (use of gloves, lab coat, safety goggles, adequate ventilation) should be followed when handling DCAF1 binder 1. Avoid inhalation, skin contact, and ingestion. The compound should be stored at -20degC in a sealed container, protected from light and moisture. DCAF1 binder 1 is for research use only and is not for human therapeutic, diagnostic, or clinical use. It is not approved by any regulatory agency.
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| References | |
| Additional Infomation |
E3 ubiquitin ligases are key components of the ubiquitin-proteasome system that control protein degradation. The CRL4 (Cullin-4 RING E3 ligase) family is one of the largest and most versatile E3 ligase families in humans. DCAF1 (DDB1- and CUL4-associated factor 1) is a substrate recognition receptor for the CRL4 complex. DCAF1 was originally identified as a binding partner of HIV-1 Vpr protein (viral protein R), which induces G2 cell cycle arrest by targeting a host protein for degradation. DCAF1 binds DDB1 and CUL4A/B to form the CRL4-DCAF1 complex, which ubiquitinates substrates including DNA damage response proteins, histones (H3 and H4), and other regulatory proteins. DCAF1 is overexpressed in certain cancers and is a potential therapeutic target. However, DCAF1 binder 1 is not used as a therapeutic agent on its own; rather, it is a chemical tool for the design of PROTACs that recruit the CRL4-DCAF1 E3 ligase to degrade disease-relevant targets (e.g., BRD4, BRD9, BTK, EGFR, etc.). Compared to the widely used E3 ligase recruiters (CRBN binders, VHL binders, IAP binders), DCAF1-recruiting PROTACs offer an alternative degradation pathway and may overcome resistance associated with mutations in CRBN or VHL pathways. DCAF1 binder 1 is a valuable addition to the chemical biology toolbox for targeted protein degradation. It is also known as a ligand for E3 ligase for targeted protein degradation. The CAS number has not been disclosed for this research compound. DCAF1 binder 1 is for research use only.
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| Molecular Formula |
C28H35CLN6O
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| Molecular Weight |
507.07
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| Appearance |
White to off-white 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) |
DMSO :~50 mg/mL (~98.61 mM)
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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 | 1.9721 mL | 9.8606 mL | 19.7211 mL | |
| 5 mM | 0.3944 mL | 1.9721 mL | 3.9442 mL | |
| 10 mM | 0.1972 mL | 0.9861 mL | 1.9721 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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