| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| Other Sizes |
| Targets |
VHL
ARD-61 is a PROTAC molecule that specifically targets the Androgen Receptor (AR) for degradation. It also effectively induces the degradation of the progesterone receptor (PR) in AR+ cancer cell lines. By recruiting an E3 ubiquitin ligase to AR and PR, it triggers their ubiquitination and subsequent destruction by the proteasome. |
|---|---|
| ln Vitro |
In order to recruit AR protein to cullin 2 for ubiquitination, followed by proteasome-dependent AR degradation, ARD-61 binds to AR protein through its AR antagonist portion and von Hippel-Lindau (VHL)/cullin 2 E3 ligase through its VHL ligand portion[1]. ARD-61 (0.001-100 μM; for 7 days) has IC50 values of 235 nM and 121 nM in the MDA-MB-453 and HCC1428 cell lines, which have the highest AR expression, respectively. With IC50 values of 39, 147, and 380 nM, respectively, ARD-61 exhibits partial inhibition of cell growth in the MCF-7, BT-549, and MDA-MB-415 cell lines, all of which have a moderate level of AR protein[1]. In all three of these AR+ breast cancer cell lines, ARD-61 (25–1000 nM; 6-72 h) induces G2/M cell cycle arrest in a dose- and time-dependent manner[1]. The MDA-MB-453 and HCC1428 cell lines undergo apoptosis when exposed to ARD-61 (25–1000000 nM; 72 h)[1]. The potency and effectiveness of ARD-61 (0.01-1000 nM; 6 h) in lowering AR protein levels is very high. In the T47D cells, ARD-61 (0.01-1000 nM; 6 h) nM; 24 h) lowers the level of PR protein with a DC50 value of 0.15 nM. The ER and GR proteins are not visibly affected by ARD-61[1]. ARD-61 (1 µM) blocks the MYC and Wnt/β-catenin signaling pathways efficiently over a 24-hour period. In addition to decreasing phosphorylated and unphosphorylated HER2 and HER3 proteins, ARD-61 (1-1000 nM) is also effective against both for a 24-hour period[1]. Both MDA-MB-453 and MCF-7 cell lines exhibit complete resistance to AR degradation induced by ARD-61 (100 nM; 24 h) when VHL is effectively knocked down[1].
In vitro, ARD-61 (0.01-1000 nM; 6 h) is highly potent and effective in reducing AR protein levels in AR+ cancer cell lines. It potently and effectively induces AR and progesterone receptor (PR) degradation, leading to the suppression of downstream signaling and the induction of apoptosis. |
| ln Vivo |
In male SCID mice, ARD-61 (25, 50 mg/kg/day; ip; for 75 days) efficiently suppresses tumor growth in the MDA-MB-453 xenograft tumor model[1]. The AR protein in the MDA-MB-453 xenograft tissue is quickly and efficiently reduced by ARD-61 (25 mg/kg; ip; for 75 days; single dose), with the effect lasting for at least 24 hours. In a time-dependent way, ARD-61 is particularly effective in lowering the mRNA level of WNT7B[1].
Cellular assays confirm that ARD-61 effectively induces apoptosis. In vivo, ARD-61 has been shown to be effective in inhibiting tumor growth in a mouse MDA-MB-453 xenograft model. By degrading AR and PR, it disrupts the survival signals in cancer cells, leading to tumor regression. |
| Enzyme Assay |
A cell-free ternary complex assay is performed using purified His-tagged AR and biotinylated VHL E3 ligase. Streptavidin donor beads and nickel chelate acceptor beads are added. Upon PROTAC-mediated binding of AR to VHL, the beads come into proximity, generating a luminescent signal. The EC50 for ternary complex formation is calculated.
|
| Cell Assay |
Cell Viability Assay[1]
Cell Types: MDA-MB-453 and HCC1428 cell lines Tested Tested Concentrations: 0.001, 0.01, 0.1, 1, 10, 100 μM Incubation Duration: 7 days Experimental Results: Achieves near complete inhibition of cell growth. Cell Cycle Analysis[1] Cell Types: MDA-MB-453, HCC1428 and MCF-7 cell lines Tested Tested Concentrations: 25, 250, 500, 1000, 10000, 100000 nM Incubation Duration: 6-72 hrs (hours) Experimental Results: Induced G2/M cell cycle arrest in a dose- and time-dependent manner in each of these three AR+ breast cancer cell lines. Apoptosis Analysis[1] Cell Types: MDA-MB-453 and HCC1428 cell lines Tested Tested Concentrations: 25, 250, 500, 1000, 10000, 100000 nM Incubation Duration: 6-72 hrs (hours) Experimental Results: Induced apoptosis in the MDA -MB-453 and HCC1428 cell lines in a dose-dependent manner. Western Blot Analysis[1] Cell Types: MDA-MB-453, MCF-7, BT549, MDA-MB-415 and HCC1428 cell lines Tested Tested Concentrations: 0.01, 0.03 , 0.1, 0.3, 1, 3, 10, 30, 100, 300, 1000 nM Incubation Duration: 6 hrs (hours) Experimental Results: diminished AR protein levels in the MDA-MB-453 (DC50=0.44 nM), MCF-7 (DC50= 1.8 Cellular degradation assays are performed in AR+ breast (MDA-MB-453) or prostate (LNCaP, VCaP) cancer cells. Cells are treated with a concentration range of ARD-61 for 6 hours. Cells are harvested, lysed, and AR and PR protein levels are measured by Western blot. The half-maximal degradation concentration (DC50) is determined. Apoptosis is confirmed via PARP and Caspase-3 cleavage by Western blot. |
| Animal Protocol |
Animal/Disease Models: MDA-MB-453 xenograft tumor model in male SCID mice[1]
Doses: 25, 50 mg/kg Route of Administration: IP; daily; for 75 days Experimental Results: Effectively inhibited tumor growth. Animal efficacy studies are conducted in mice bearing MDA-MB-453 xenografts. ARD-61 is administered via intraperitoneal or intravenous injection. Tumor volumes are measured with calipers. At study termination, tumors are harvested for Western blot analysis to confirm degradation of AR and PR protein in vivo and to assess markers of apoptosis. |
| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of ARD-61 are currently being evaluated to support its in vivo use. As a large PROTAC molecule (MW 1095.78), optimizing its absorption and tissue distribution is a key focus. Studies are ongoing to determine its half-life and bioavailability in rodent models.
|
| Toxicity/Toxicokinetics |
Toxicological data is limited as ARD-61 is a highly potent research tool. Its safety is primarily evaluated in cell-based assays, where it shows activity. In vivo, the primary toxicities are likely to be on-target effects related to the potent degradation of AR and PR in normal tissues. Long-term safety assessments are not standard for this reagent.
|
| References | |
| Additional Infomation |
ARD-61 is a PROTAC with the molecular formula C61H71ClN8O7S and a molecular weight of 1095.78. It is a highly specific chemical probe for depleting AR and PR proteins, providing researchers with a powerful tool to dissect the consequences of acute protein loss, which is distinct from the effects of transcriptional or translational inhibition.
|
| Molecular Formula |
C61H71CLN8O7S
|
|---|---|
| Molecular Weight |
1095.78385281563
|
| Exact Mass |
1094.485
|
| CAS # |
2316837-08-6
|
| PubChem CID |
154704831
|
| Appearance |
White to light yellow solid powder
|
| LogP |
9.3
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
12
|
| Rotatable Bond Count |
15
|
| Heavy Atom Count |
78
|
| Complexity |
2200
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
ClC1=C(C#N)C=CC(=C1)OC1C(C)(C)C(C1(C)C)NC(C1C=CC(C#CC2CCN(CC2)C2CCN(C(C[C@@H](C3C=CC(C4=C(C)N=CS4)=CC=3)NC([C@@H]3C[C@H](CN3C([C@@H](C3=CC(C)=NO3)C(C)C)=O)O)=O)=O)CC2)=CC=1)=O
|
| InChi Key |
RGPUSLXMPBTXNU-CUPIBCLHSA-N
|
| InChi Code |
InChI=1S/C61H71ClN8O7S/c1-36(2)53(51-29-37(3)67-77-51)57(75)70-34-46(71)30-50(70)56(74)65-49(41-15-17-42(18-16-41)54-38(4)64-35-78-54)32-52(72)69-27-23-45(24-28-69)68-25-21-40(22-26-68)10-9-39-11-13-43(14-12-39)55(73)66-58-60(5,6)59(61(58,7)8)76-47-20-19-44(33-63)48(62)31-47/h11-20,29,31,35-36,40,45-46,49-50,53,58-59,71H,21-28,30,32,34H2,1-8H3,(H,65,74)(H,66,73)/t46-,49+,50+,53-,58?,59?/m1/s1
|
| Chemical Name |
(2S,4R)-N-[(1S)-3-[4-[4-[2-[4-[[3-(3-chloro-4-cyanophenoxy)-2,2,4,4-tetramethylcyclobutyl]carbamoyl]phenyl]ethynyl]piperidin-1-yl]piperidin-1-yl]-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]-3-oxopropyl]-4-hydroxy-1-[(2R)-3-methyl-2-(3-methyl-1,2-oxazol-5-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 (91.26 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (2.28 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 25.0 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: ≥ 2.5 mg/mL (2.28 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 25.0 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: ≥ 2.5 mg/mL (2.28 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9126 mL | 4.5630 mL | 9.1259 mL | |
| 5 mM | 0.1825 mL | 0.9126 mL | 1.8252 mL | |
| 10 mM | 0.0913 mL | 0.4563 mL | 0.9126 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
