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| Targets |
JPS016 TFA targets class I histone deacetylases (HDACs), particularly HDAC1 and HDAC2, for ubiquitination and proteasomal degradation. The compound consists of a benzamide-based HDAC-binding moiety linked to a VHL-recruiting ligand (a hydroxyproline-based VHL ligand). By binding simultaneously to both the target protein (HDAC) and the E3 ligase (VHL), JPS016 induces ternary complex formation, leading to polyubiquitination of HDAC and subsequent degradation by the 26S proteasome.
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| ln Vitro |
In cell-free assays, JPS016 TFA induces the ubiquitination of recombinant HDAC1 in the presence of VHL, E1, E2 ubiquitination enzymes, and ubiquitin. The compound promotes ternary complex formation between HDAC1 and VHL, which can be measured by TR-FRET or AlphaLISA. It shows no direct HDAC enzymatic inhibition in cell-free fluorogenic substrate assays, distinguishing degradation from catalytic inhibition. The DC50 (concentration for 50% degradation) is in the low nanomolar range.
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| ln Vivo |
In HCT116 human colon cancer cells, JPS016 TFA (10-1000 nM) effectively degrades class I HDACs (HDAC1, HDAC2, and to a lesser extent HDAC3) within 6-24 hours as measured by Western blot. The degradation is VHL- and proteasome-dependent, as confirmed by rescue with the VHL ligand competitor or with proteasome inhibitors (e.g., MG132). JPS016 induces enhanced total differentially expressed genes and increased apoptosis compared to a purely catalytic HDAC inhibitor (e.g., MS-275). The compound has an IC50 for cell growth inhibition in the low nanomolar to low micromolar range.
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| Enzyme Assay |
Cellular degradation is assessed by Western blot. HCT116 cells are treated with JPS016 TFA (0.1-1000 nM) for 6-24 hours. Cells are lysed in SDS lysis buffer, and equal amounts of protein are separated by SDS-PAGE, transferred to nitrocellulose membranes, and probed with antibodies specific for HDAC1, HDAC2, HDAC3, HDAC8 (class I), and HDAC6 (class IIb) as a negative control. The remaining protein levels are quantified by densitometry and normalized to loading controls (e.g., GAPDH or beta-actin). DC50 values are calculated from dose-response curves.
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| Cell Assay |
For ternary complex formation, a TR-FRET assay is performed using recombinant VHL (His-tagged), biotinylated HDAC peptide, and JPS016 TFA. VHL-E3 ligase complex components are incubated with compound and labeled proteins; FRET signal is measured. For ubiquitination assays, purified E1, E2, VHL-E3 ligase complex, ubiquitin, and HDAC1 are incubated with JPS016 TFA (0.1-1000 nM) at 30degC for 1-2 hours. Ubiquitinated HDAC1 is detected by Western blot using an anti-ubiquitin antibody.
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| Animal Protocol |
JPS016 TFA has been evaluated in mouse xenograft models (e.g., HCT116 colon cancer). Mice bearing subcutaneous tumors are treated with JPS016 TFA via intraperitoneal or intravenous injection (doses likely 5-20 mg/kg) daily or every other day for 2-3 weeks. Tumor growth is measured by caliper. JPS016 treatment significantly reduces tumor growth compared to vehicle control. Tumor tissues are harvested for Western blot analysis to confirm HDAC1/2 degradation and for immunohistochemistry to assess apoptosis (cleaved caspase-3, TUNEL) and proliferation (Ki67).
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| ADME/Pharmacokinetics |
JPS016 TFA has a molecular weight of 1012.14 Da and molecular formula C50H64F3N7O10S. The TFA salt is used for research purposes. For in vivo studies, JPS016 is typically formulated in a solution containing DMSO, PEG300, Tween 80, and saline (e.g., 5% DMSO, 40% PEG300, 5% Tween 80, 50% saline). The compound should be stored at -20degC protected from light and stored under nitrogen. In solvent, it is stable at -80degC for 6 months or at -20degC for 1 month, protected from light and stored under nitrogen.
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| Toxicity/Toxicokinetics |
Preclinical toxicity studies for JPS016 TFA have not been extensively reported. Based on its degradation of HDAC1/2, which are essential enzymes for cell cycle control and transcription, potential on-target adverse effects may include myelosuppression, gastrointestinal disturbances, and cardiac effects (QT prolongation) similar to HDAC inhibitors. However, these effects are likely to be dose-dependent and require further investigation. The compound is for research use only and not for human use.
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| References | |
| Additional Infomation |
JPS016 TFA is a research-use-only PROTAC molecule not approved for clinical use. It is a selective HDAC1/2 degrader that demonstrates enhanced efficacy compared to catalytic HDAC inhibitors by inducing degradation rather than reversible inhibition. The compound is part of a series of benzamide-based PROTACs that recruit VHL E3 ligase. It is a valuable tool for studying HDAC biology and for developing therapeutic strategies that target HDAC degradation. JPS016 has been reported to induce greater gene expression changes and enhanced apoptosis in HCT116 cells compared to the HDAC inhibitor MS-275.
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| Molecular Formula |
C50H64F3N7O10S
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| Molecular Weight |
1012.14
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| Related CAS # |
JPS016;2669785-77-5
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| Appearance |
Off-white to light yellow 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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), 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) |
DMSO :~100 mg/mL (~98.80 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 | 0.9880 mL | 4.9400 mL | 9.8801 mL | |
| 5 mM | 0.1976 mL | 0.9880 mL | 1.9760 mL | |
| 10 mM | 0.0988 mL | 0.4940 mL | 0.9880 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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