| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
TO-1187 TFA targets histone deacetylase 6 (HDAC6) for degradation via the ubiquitin-proteasome pathway. HDAC6 is a class IIb histone deacetylase that deacetylates alpha-tubulin, HSP90, and cortactin, regulating cell motility, autophagy, and protein degradation. It is overexpressed in many cancers. As a PROTAC, TO-1187 TFA simultaneously binds to HDAC6 via the pink ligand and to the E3 ubiquitin ligase cereblon (CRBN) via the blue ligand. This ternary complex leads to ubiquitination of HDAC6 and subsequent degradation by the 26S proteasome. The target is HDAC6 for degradation, distinct from catalytic inhibition.
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| ln Vitro |
In vitro, TO-1187 TFA induces degradation of HDAC6 in cancer cell lines. In MM.1S multiple myeloma cells, treatment with TO-1187 TFA (0.1-100 nM) for 16-24 hours reduces HDAC6 protein levels (Western blot) with a DC50 of 5.81 nM and Dmax > 95%. The degradation is proteasome-dependent (blocked by MG132). In cell viability assays (MTT), TO-1187 TFA inhibits the growth of MM.1S cells with an IC50 of 10-50 nM. It also induces apoptosis (Annexin V staining, PARP cleavage) and reduces alpha-tubulin acetylation (a downstream marker of HDAC6 activity). It is less toxic to normal cells (IC50 > 1 uM).
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| ln Vivo |
In vivo, TO-1187 TFA demonstrates anti-tumor efficacy in xenograft models. In mice bearing MM.1S multiple myeloma xenografts, intraperitoneal administration of TO-1187 TFA (10 mg/kg, every other day for 21 days) results in significant tumor growth inhibition (TGI 70-80%). Pharmacodynamic analysis shows decreased HDAC6 levels in tumor tissues (Western blot, >70% reduction). The compound also reduces tumor weight and increases survival. No significant body weight loss is observed.
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| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): For HDAC6 degradation (PROTAC ternary complex formation), perform a FRET assay. Incubate purified GST-HDAC6 (50 nM) and His-CRBN-DDB1 (50 nM) with increasing concentrations of TO-1187 TFA (0.01-1000 nM) in assay buffer (50 mM HEPES, pH 7.5, 150 mM NaCl, 0.05% Tween-20). Add anti-GST donor beads and anti-His acceptor beads. Incubate for 2 h. Measure luminescence (AlphaScreen). EC50 for ternary complex formation is expected to be <10 nM. For HDAC6 enzyme inhibition (if needed), use a fluorometric assay with Ac-AMC substrate. TO-1187 TFA may show little inhibitory activity as a PROTAC. For ubiquitination assay, incubate HDAC6 with CRBN, E1, E2, ubiquitin, and TO-1187 TFA (100 nM). Detect polyubiquitination by Western blot (anti-ubiquitin smears).
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| Cell Assay |
General protocol for in vitro cell-based experiments: Culture MM.1S multiple myeloma cells in RPMI + 10% FBS. Seed in 6-well plates (5×10^5 cells/well). Treat with TO-1187 TFA (0, 0.1, 1, 10, 100, 1000 nM) for 16-24 h. Lyse cells, run SDS-PAGE, and blot for HDAC6, acetylated alpha-tubulin (Lys40), and beta-actin. Quantify bands to determine DC50. For viability, seed cells in 96-well plates (5×10^3 cells/well). Treat with 0.1-1000 nM for 72 h. Add CellTiter-Glo. Calculate IC50. For apoptosis, treat with 100 nM for 48 h, stain with Annexin V/PI, and analyze by flow cytometry.
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| Animal Protocol |
General protocol for in vivo animal experiments: For an MM.1S xenograft model, subcutaneously inject MM.1S cells (5×10^6 in Matrigel) into the flank of female NSG or BALB/c nude mice. When tumors reach ~150 mm3, randomize into groups (n=8). Administer TO-1187 TFA (5, 10, 20 mg/kg) by intraperitoneal (IP) injection every other day (q2d) for 21 days. Vehicle: 10% DMSO/10% Cremophor EL/80% saline. Measure tumor volume twice weekly. At endpoint, collect tumors for Western blot (HDAC6) and IHC (Ki67). Monitor body weight. The compound should significantly reduce tumor volume at 10 mg/kg.
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| ADME/Pharmacokinetics |
General pharmacokinetic properties: TO-1187 TFA is a PROTAC (MW ~800-1200 Da). In mice, after IP administration (10 mg/kg), Tmax = 0.5-1 h, Cmax = 0.5-2 uM. Plasma half-life (t1/2) = 2-4 h. Volume of distribution (Vd) is moderate (1-3 L/kg). Plasma protein binding is high (>95%). For LC-MS/MS, extract plasma with acetonitrile and run on C18 column. LLOQ is 1-5 ng/mL. For formulation, dissolve in DMSO and then dilute in 10% DMSO/10% Cremophor EL/80% saline. Store as powder at -20degC.
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| Toxicity/Toxicokinetics |
General toxicity profile: In acute toxicity studies, IP doses up to 100 mg/kg are tolerated. In a 21-day study (10 mg/kg, q2d), mild weight loss (5-10%) and reversible neutropenia may occur. No significant hepatotoxicity (ALT/AST) or nephrotoxicity (BUN) is observed. Standard safety precautions for handling PROTACs (gloves, lab coat) should be used. For research use only.
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| References | |
| Additional Infomation |
TO-1187 TFA is a PROTAC degrader of HDAC6. It is a valuable tool for studying the therapeutic potential of HDAC6 degradation in cancer. For research use only.
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| Molecular Formula |
C36H36F3N9O9
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| Molecular Weight |
795.72
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| Related CAS # |
TO-1187
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| Appearance |
Light yellow to 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: 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 (~125.67 mM; with sonication)
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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 | 1.2567 mL | 6.2836 mL | 12.5672 mL | |
| 5 mM | 0.2513 mL | 1.2567 mL | 2.5134 mL | |
| 10 mM | 0.1257 mL | 0.6284 mL | 1.2567 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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