| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
VHL
Von Hippel-Lindau (VHL) E3 ubiquitin ligase. This compound is a high-affinity VHL ligand that recruits the VHL protein, a component of the E3 ubiquitin ligase complex, for targeted protein degradation applications in PROTAC technology. |
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| ln Vitro |
As a VHL ligand, (S,R,S)-AHPC-Me binds to the VHL protein with high affinity (KD in the low nM range, similar to the parent (S,R,S)-AHPC). It is used as a building block for PROTAC synthesis. For example, ARV-771 is a BET PROTAC degrader based on VHL E3 ligase that efficiently degrades BET protein in castration-resistant prostate cancer (CRPC) cells with a DC50 < 1 nM.
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| ln Vivo |
No direct in vivo activity. When incorporated into a PROTAC molecule such as ARV-771, the VHL ligand enables the PROTAC to recruit the VHL E3 ligase to target proteins, leading to their ubiquitination and proteasomal degradation in vivo. ARV-771 (10-100 mg/kg, SC, daily) has shown efficacy in CRPC xenograft models.
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| Enzyme Assay |
(1) VHL binding assay: Perform surface plasmon resonance (SPR) or fluorescence polarization (FP) using purified recombinant VHL-elongin B-elongin C complex. (2) Immobilize VHL complex on a sensor chip. (3) Inject (S,R,S)-AHPC-Me dihydrochloride at concentrations ranging from 0.1 nM to 10 uM. (4) Measure binding kinetics to determine KD. (5) Alternatively, use TR-FRET competition assay with a fluorescently labeled VHL ligand.
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| Cell Assay |
(1) For direct cellular activity: None; the compound alone does not have cellular activity. (2) For PROTAC cellular assays: treat cells with a complete PROTAC (e.g., ARV-771) synthesized using this VHL ligand (0.1-1000 nM, 4-24 h). (3) Lyse cells, perform SDS-PAGE, and Western blot with antibodies against the target protein (e.g., BET family proteins BRD2, BRD3, BRD4). (4) Quantify target protein degradation by densitometry normalized to loading control. (5) Determine DC50 (concentration for 50% degradation) and Dmax (maximum degradation).
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| Animal Protocol |
No in vivo animal protocol for the ligand alone. For the complete PROTAC (e.g., ARV-771): (1) 6-8 week old male BALB/c nude mice bearing CRPC xenografts (22Rv1 or VCaP cells, 200-300 mm3). (2) Administer ARV-771 subcutaneously (SC) at 10-100 mg/kg daily or every other day. (3) Formulation: 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. (4) Monitor tumor volume (caliper) and body weight for 14-28 days. (5) Harvest tumors 4-24 h after final dose, perform Western blot for BET proteins and IHC for Ki-67 and cleaved caspase-3.
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| ADME/Pharmacokinetics |
Standard formulation for the ligand: dissolve in DMSO as a 50-100 mM stock solution. For storage: powder at -20degC for 3 years; in solvent at -80degC for 6 months. For PROTAC in vivo formulation: prepare 10-20 mg/mL stock in DMSO, then dilute in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline for IP/SC injection. PK of ARV-771 in mice: SC (50 mg/kg), Cmax ~ 5-10 uM, t1/2 ~ 2-4 h, AUC ~ 10-30 uM·h.
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| Toxicity/Toxicokinetics |
For in vitro toxicity of the ligand: CCK-8 assay on HEK293 cells, IC50 > 100 uM. For in vivo toxicity of the PROTAC: MTD studies in CD-1 mice, single SC dose up to 200 mg/kg; repeated dosing at 50-100 mg/kg SC daily for 14 days, monitor body weight, clinical signs, and clinical chemistry (ALT, AST, BUN, creatinine). The ligand is for research use only, not for human use.
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| References | |
| Additional Infomation |
(S,R,S)-AHPC-Me dihydrochloride is a VHL ligand that recruits the von Hippel-Lindau (VHL) protein for PROTAC-based protein degradation. It may be utilized to prepare ARV-771, a BET PROTAC degrader based on VHL E3 ligase. ARV-771 is the first reported BET PROTAC with in vivo efficacy in CRPC models. This product is for research use only, not for human use.
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| Molecular Formula |
C23H34CL2N4O3S
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|---|---|
| Molecular Weight |
517.512062549591
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| Exact Mass |
516.172
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| CAS # |
2504950-56-3
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| Related CAS # |
(S,R,S)-AHPC-Me;1948273-02-6;(S,R,S)-AHPC-Me hydrochloride;1948273-03-7
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| PubChem CID |
137319712
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
33
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| Complexity |
647
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CC1=C(SC=N1)C2=CC=C(C=C2)[C@H](C)NC(=O)[C@@H]3C[C@H](CN3C(=O)[C@H](C(C)(C)C)N)O.Cl.Cl
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| InChi Key |
YTJDUDNKHHNDRD-ARZBPYMHSA-N
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| InChi Code |
InChI=1S/C23H32N4O3S.2ClH/c1-13(15-6-8-16(9-7-15)19-14(2)25-12-31-19)26-21(29)18-10-17(28)11-27(18)22(30)20(24)23(3,4)5;;/h6-9,12-13,17-18,20,28H,10-11,24H2,1-5H3,(H,26,29);2*1H/t13-,17+,18-,20+;;/m0../s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-amino-3,3-dimethylbutanoyl]-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide;dihydrochloride
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
H2O :~100 mg/mL (~193.23 mM)
DMSO :~50 mg/mL (~96.62 mM ) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (9.66 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 5 mg/mL (9.66 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 50.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 5 mg/mL (9.66 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 100 mg/mL (193.23 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9323 mL | 9.6616 mL | 19.3233 mL | |
| 5 mM | 0.3865 mL | 1.9323 mL | 3.8647 mL | |
| 10 mM | 0.1932 mL | 0.9662 mL | 1.9323 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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