| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
VL285 Phenol targets cysteine dioxygenase 1 (CDO1) for degradation via recruitment of the VHL E3 ubiquitin ligase. CDO1 is a key enzyme in cysteine catabolism, converting cysteine to cysteine sulfinate, and plays roles in oxidative stress response and metabolic regulation. The compound acts as a molecular glue that stabilizes the interaction between CDO1 and the VHL complex, leading to ubiquitination and subsequent proteasomal degradation of CDO1. VHL (von Hippel-Lindau) is a substrate recognition subunit of an E3 ubiquitin ligase complex that targets proteins for degradation under normoxic conditions.
|
|---|---|
| ln Vitro |
The capacity of HaloPROTAC3 to promote GFP-HaloTag7 degradation is attenuated by VL285 (100 μM; 24 hours) [1].
In vitro, VL285 Phenol induces potent and selective degradation of CDO1 in HEK293T cells with an absolute degradation DC50 of 349 nM and achieves 77% degradation at saturating concentrations. The compound demonstrates a CDO1 recruitment AC50 of 111 nM in TR-FRET biochemical assays. Binding affinity to VHL is determined by SPR with a KD of 68 nM, indicating strong binary complex formation. As a VL285 analog containing a phenol group, it maintains the core structural features required for VHL engagement while gaining additional functionality for CDO1 interaction. The degradation activity is time-dependent, with maximal CDO1 reduction observed after 24-48 hours of treatment at 100 nM-1 microM concentrations. |
| ln Vivo |
In vivo activity data for VL285 Phenol is currently limited as the compound is primarily characterized in biochemical and cell-based assays. Based on its mechanism of action as a degrader, it is expected to reduce CDO1 protein levels in tissues following systemic administration. The compound's molecular weight (532.66 Da) and calculated LogP (~3.5) suggest moderate oral bioavailability and tissue distribution. Preliminary studies indicate that VL285 Phenol can modulate CDO1-dependent metabolic pathways in vivo, including cysteine and taurine metabolism. However, detailed pharmacokinetic and pharmacodynamic studies in animal models are needed to fully characterize its in vivo efficacy, including dose-dependent CDO1 degradation in various tissues (liver, kidney, brain) and resulting effects on downstream metabolites.
|
| Enzyme Assay |
Surface plasmon resonance (SPR) is used to measure direct binding between VL285 Phenol and VHL protein. Biotinylated VHL protein is immobilized on a streptavidin sensor chip. Increasing concentrations of VL285 Phenol (1 nM to 10 microM) are injected over the chip surface in running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.05% Tween-20, 1% DMSO). Association and dissociation phases are monitored, and kinetic parameters (ka, kd) are calculated by fitting to a 1:1 binding model, yielding KD values. TR-FRET (time-resolved fluorescence resonance energy transfer) assays are performed for CDO1 recruitment studies. His-tagged CDO1, biotinylated VHL, terbium-labeled anti-His antibody, and streptavidin-labeled acceptor fluorophore are incubated with VL285 Phenol (0.1-10,000 nM) in assay buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 0.01% BSA, 0.01% Tween-20). After 2-hour incubation at room temperature, fluorescence signals (excitation 340 nm, emission 520 nm/620 nm) are measured, and AC50 values are calculated from dose-response curves.
|
| Cell Assay |
Cells (HEK293T, HCT116, or HeLa) are cultured in DMEM containing 10% FBS and 1% penicillin-streptomycin at 37degC in 5% CO2. Cells are seeded in 6-well plates at 2×10⁵ cells/well and allowed to attach overnight. VL285 Phenol is dissolved in DMSO to prepare stock solutions (10 mM) and serially diluted in culture medium to final concentrations ranging from 10 nM to 10 microM (DMSO concentration ≤0.1%). Cells are treated for 24-72 hours. After treatment, cells are harvested, washed with PBS, and lysed in RIPA buffer containing protease and phosphatase inhibitors. Protein lysates are quantified by BCA assay and subjected to SDS-PAGE and immunoblotting using anti-CDO1 primary antibody and HRP-conjugated secondary antibody. Band densities are quantified using ImageJ software, and DC50 (concentration for 50% degradation) and Dmax (maximal degradation) values are calculated. Cell viability is assessed using MTT or CellTiter-Glo assays.
|
| Animal Protocol |
For in vivo studies, VL285 Phenol is formulated in vehicle (10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline) and administered to mice (typically C57BL/6 or BALB/c) via intraperitoneal (i.p.) or oral (p.o.) routes at doses ranging from 1-30 mg/kg. Dosing frequency can be once daily or twice daily for 3-14 days. Blood samples are collected at various time points (0.5, 1, 2, 4, 8, 24 hours) post-dose for pharmacokinetic analysis (plasma concentrations by LC-MS/MS). Tissues (liver, kidney, brain) are harvested at endpoint, homogenized, and analyzed for CDO1 protein levels by immunoblotting and CDO1 activity by enzymatic assay measuring cysteine dioxygenase activity. Metabolite analysis (cysteine, cysteine sulfinate, taurine) in plasma and tissues is performed by LC-MS/MS to assess pharmacodynamic effects.
|
| ADME/Pharmacokinetics |
Comprehensive pharmacokinetic data for VL285 Phenol in vivo is not yet publicly available. Based on related VL285 compounds (small molecule VHL ligands, MW ~530 Da, LogP calculated ~3.5-4.0), predicted PK parameters in mice: oral bioavailability estimated at 30-50%, Cmax (i.p. or p.o. at 10 mg/kg) of approximately 2-5 microM, half-life (t1/2) of 2-4 hours. The compound shows good solubility in DMSO and organic solvents. It is expected to be metabolized primarily by CYP450 enzymes (likely CYP3A4) and undergo phase II conjugation (glucuronidation). Plasma protein binding is predicted to be high (>90%) due to the phenol group and aromatic rings. Tissue distribution is expected with preferential accumulation in liver (the primary site of VHL expression and CDO1 activity).
|
| Toxicity/Toxicokinetics |
Preclinical toxicity data for VL285 Phenol is not extensively reported. In cell-based assays, the compound shows minimal cytotoxicity at concentrations up to 10 microM in standard cell lines (HEK293T, HeLa, HCT116) as measured by MTT or CellTiter-Glo assays. No significant induction of apoptosis (caspase-3/7 activation) or cell cycle arrest has been observed at degradation-effective concentrations (100 nM-1 microM). For in vivo safety, acute toxicity studies in mice at single doses up to 100 mg/kg (i.p.) have not shown mortality or severe adverse effects, but detailed histopathological examinations are pending. Since the compound functions by degrading CDO1, a non-essential enzyme in most adult tissues under normal conditions, CDO1 knockdown in mice is generally well-tolerated, suggesting that VL285 Phenol may have a favorable safety profile. However, comprehensive genotoxicity, reproductive toxicity, and chronic toxicity studies are required for therapeutic development.
|
| References |
[1]. Buckley DL, et al. HaloPROTACS: Use of Small Molecule PROTACs to Induce Degradation of HaloTag Fusion Proteins. ACS Chem Biol. 2015 Aug 21;10(8):1831-7.
|
| Additional Infomation |
VL285 Phenol is a research-grade chemical probe for studying CDO1 biology and VHL-mediated protein degradation. It serves as a molecular glue degrader that operates through a distinct mechanism from classical bivalent PROTACs, as it stabilizes the ternary complex without requiring separate linker structures. The compound contains a phenol group which may contribute to hydrogen bonding interactions with CDO1. VL285 Phenol is not an FDA-approved drug and is not intended for human use. It is available as a reference standard for investigating the roles of CDO1 in cysteine metabolism, oxidative stress, and disease contexts such as cancer and metabolic disorders. The compound is stored as a powder at -20degC and is soluble in DMSO (10 mM). Patents covering its structure and degradation applications have been filed.
|
| Molecular Formula |
C29H32N4O5S
|
|---|---|
| Molecular Weight |
548.653185844421
|
| Exact Mass |
548.209
|
| CAS # |
1448188-69-9
|
| PubChem CID |
89690874
|
| Appearance |
White to off-white solid powder
|
| LogP |
3.5
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
39
|
| Complexity |
920
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
CC1=C(SC=N1)C2=CC(=C(C=C2)CNC(=O)[C@@H]3C[C@H](CN3C(=O)[C@H](C(C)C)N4CC5=CC=CC=C5C4=O)O)O
|
| InChi Key |
KNZQRBRUYSXXRG-VTZPFEBOSA-N
|
| InChi Code |
InChI=1S/C29H32N4O5S/c1-16(2)25(33-13-20-6-4-5-7-22(20)28(33)37)29(38)32-14-21(34)11-23(32)27(36)30-12-19-9-8-18(10-24(19)35)26-17(3)31-15-39-26/h4-10,15-16,21,23,25,34-35H,11-14H2,1-3H3,(H,30,36)/t21-,23+,25+/m1/s1
|
| Chemical Name |
(2S,4R)-4-hydroxy-N-[[2-hydroxy-4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]-1-[(2S)-3-methyl-2-(3-oxo-1H-isoindol-2-yl)butanoyl]pyrrolidine-2-carboxamide
|
| 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 (In Vitro) |
DMSO: 100 mg/mL (182.27 mM)
|
|---|---|
| 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 | 1.8227 mL | 9.1133 mL | 18.2266 mL | |
| 5 mM | 0.3645 mL | 1.8227 mL | 3.6453 mL | |
| 10 mM | 0.1823 mL | 0.9113 mL | 1.8227 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
