| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
SUV39H2/KMT1B
OTS186935 hydrochloride specifically targets SUV39H2 (suppressor of variegation 3-9 homolog 2), a lysine methyltransferase that catalyzes the di- and tri-methylation of histone H3 at lysine 9 (H3K9me3). H3K9me3 is a repressive histone mark associated with heterochromatin formation, transcriptional silencing, and genome stability. SUV39H2 is often overexpressed in various human cancers, including lung, colon, and breast cancers, where it contributes to tumor progression and poor prognosis. By inhibiting SUV39H2, OTS186935 hydrochloride reduces H3K9me3 levels, leading to transcriptional reactivation of silenced tumor suppressor genes and induction of DNA damage responses. |
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| ln Vitro |
At an IC50 of 0.67 μM, OTS186935 hydrochloride inhibits the proliferation of A549 cells[1].
In vitro, OTS186935 hydrochloride potently inhibits the proliferation of A549 human lung adenocarcinoma cells with an IC50 of 0.67 microM. The compound reduces global H3K9me3 levels in a dose-dependent manner as measured by Western blotting and immunofluorescence. Treatment of cancer cells with OTS186935 hydrochloride (0.1-10 microM, 72 hours) induces cell cycle arrest at G2/M phase and promotes apoptosis, as indicated by increased cleaved PARP and caspase-3. The compound also regulates the phosphorylation of H2AX (gamma-H2AX), a marker of DNA double-strand breaks, suggesting that SUV39H2 inhibition leads to genomic instability in cancer cells. The IC50 for SUV39H2 enzyme inhibition is 6.49 nM, demonstrating high potency for the target. |
| ln Vivo |
In xenograft models created from human cancer cell lines, OTS186935 hydrochloride (10 mg/kg or 25 mg/kg; intravenously; once daily for 14 days) shows growth inhibition [1].
In vivo, OTS186935 hydrochloride shows significant tumor growth inhibition in mouse xenograft models without detectable toxicity. In MDA-MB-231 triple-negative breast cancer xenografts in female NOD.CB17-Prkdcscid/J mice, treatment with OTS186935 hydrochloride (10 mg/kg, intravenous, once daily for 14 days) results in 42.6% tumor growth inhibition on day 14 compared to vehicle control. In A549 lung cancer xenografts in female BALB/cAJcl-nu/nu nude mice, a higher dose of 25 mg/kg (intravenous, once daily for 14 days) yields 60.8% tumor growth inhibition without significant body weight loss or signs of toxicity. The compound also inhibits tumor growth in other xenograft models derived from human cancer cell lines. Tumor tissues from treated animals show reduced H3K9me3 levels by immunohistochemistry. |
| Enzyme Assay |
Cell-free SUV39H2 enzyme inhibition assays are performed using recombinant human SUV39H2 enzyme. The reaction mixture (50 microL) contains 50 mM Tris-HCl (pH 8.5), 10 mM DTT, 0.01% Tween-20, 1 microM biotinylated histone H3 peptide (residues 1-21), 1 microM [3H]-SAM (S-adenosylmethionine), and varying concentrations of OTS186935 hydrochloride (0.01-100 nM). The reaction is initiated by adding 10 ng SUV39H2 enzyme and incubated at 30degC for 60 minutes. The reaction is stopped by adding 50 microL of 7.5 M guanidine hydrochloride. The reaction mixture is transferred to a Streptavidin-coated FlashPlate, incubated for 60 minutes at 30degC, and counted on a scintillation counter. Alternatively, the amount of radiolabeled methyl groups incorporated into the histone peptide is quantitated by filtration onto filter mats. The IC50 (6.49 nM) is calculated from the dose-response curve.
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| Cell Assay |
A549 human lung adenocarcinoma cells are used for in vitro proliferation and mechanistic studies. Cells are seeded in 96-well plates at 5×103 cells/well and allowed to attach overnight. OTS186935 hydrochloride (0.01-100 microM) is added, and cells are incubated for 48-72 hours. Cell viability is assessed using CellTiter-Glo luminescent assay or MTT assay. IC50 (0.67 microM) is calculated by nonlinear regression. For flow cytometry analysis of cell cycle, cells are seeded in 6-well plates (2×10⁵ cells/well), treated with compound (0.1-10 microM) for 48 hours, fixed in 70% ethanol, stained with propidium iodide (50 microg/mL) containing RNase A (100 microg/mL), and analyzed on a flow cytometer. For gamma-H2AX analysis, treated cells are fixed, permeabilized, stained with anti-gamma-H2AX antibody conjugated to Alexa Fluor 488, and analyzed by flow cytometry. Western blotting is performed for H3K9me3, H3, cleaved PARP, and cleaved caspase-3.
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| Animal Protocol |
Animal/Disease Models: Female NOD.CB17-Prkdcscid/J mice (bearing MDA-MB-231 cells )[1]
Doses: 10 mg/kg Route of Administration: intravenously (iv); one time/day for 14 days Experimental Results: Tumor growth inhibition of 42.6% on day 14 . Animal/Disease Models: Female BALB/cAJcl-nu/nu (nude) mice (bearing A549 cells)[1] Doses: 25 mg/kg Route of Administration: intravenously (iv); one time/day for 14 days Experimental Results: Yielded a tumor growth inhibition of 60.8% without significant body weight loss or toxicity. For in vivo efficacy studies, 5-6 week old female NOD.CB17-Prkdcscid/J mice (for MDA-MB-231) or female BALB/cAJcl-nu/nu nude mice (for A549) are used. 5×10⁶ cancer cells in 100 microL PBS are injected subcutaneously into the right flank. When tumors reach an average volume of 150-250 mm3 (approximately day 14-21 post-inoculation), mice are randomized into treatment groups (n=8-10 per group). OTS186935 hydrochloride is formulated in a vehicle of 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline or 10% DMSO + 90% (20% SBE-beta-CD in saline). The compound is administered intravenously via the tail vein once daily at doses of 10 mg/kg (MDA-MB-231 model) or 25 mg/kg (A549 model) for 14 consecutive days. Control groups receive vehicle alone. Tumor volume is measured twice weekly with calipers and calculated as (length × width2)/2. Body weight is monitored as a surrogate marker of toxicity. At study termination (day 14 or day 28), tumors are excised, weighed, and processed for immunohistochemistry (H3K9me3, Ki-67) and Western blotting (H3K9me3, gamma-H2AX). |
| ADME/Pharmacokinetics |
OTS186935 hydrochloride has a molecular weight of 522.31 g/mol (free base) and is supplied as a light yellow to yellow solid powder. The compound should be stored as a powder at -20degC (3 years) or 4degC (2 years) in sealed, moisture-protected containers. For solution storage, it should be kept at -80degC for 6 months or -20degC for 1 month. The compound is soluble in water (∼50 mg/mL, ∼95.73 mM) and DMSO (∼25 mg/mL, ∼47.86 mM). For intravenous administration, working solutions are prepared using the sequential addition method: 100 microL of a 20.8 mg/mL DMSO stock solution is added to 400 microL PEG300, mixed well; 50 microL Tween-80 is added and mixed; then 450 microL normal saline is added to achieve a final concentration of ∼2.08 mg/mL (3.98 mM). For formulations with SBE-beta-CD, dissolve in 10% DMSO + 90% (20% SBE-beta-CD in saline).
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| Toxicity/Toxicokinetics |
This product is for research use only and is not for human therapeutic applications. No significant toxicity was observed in mouse xenograft studies at the tested doses (10 mg/kg and 25 mg/kg). Body weight loss was not significant, and no overt signs of toxicity (behavioral changes, reduced activity, ruffled fur, or diarrhea) were reported. The compound's safety profile is favorable in these preclinical models. However, as a histone methyltransferase inhibitor, off-target epigenetic effects may occur. Standard laboratory safety practices (gloves, lab coat, safety glasses) should be followed. Avoid inhalation of powder and contact with skin. The compound should be handled in a chemical fume hood. Not for human use.
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| References | |
| Additional Infomation |
SUV39H2 is a histone methyltransferase that is frequently overexpressed in various cancers and is associated with poor prognosis. It is part of the SUV39 family, which also includes SUV39H1. OTS186935 hydrochloride is a first-in-class SUV39H2 inhibitor that shows specificity for SUV39H2 over other histone methyltransferases. The regulation of gamma-H2AX production in cancer cells by SUV39H2 inhibition represents a novel mechanism for inducing cancer cell death through accumulation of DNA damage. This compound is a useful chemical probe for studying the function of SUV39H2 in cellular senescence, tumor suppression, and chromatin dynamics. The compound may also be referred to as OTS186935 trihydrochloride. OTS186935 is covered by patent literature. This is not a clinically approved drug. Supplier information must not be included.
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| Molecular Formula |
C25H26CLN5O2.8/5HCL
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| Molecular Weight |
522.31
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| Related CAS # |
OTS186935;2093400-18-9;OTS186935 trihydrochloride;2093401-85-3
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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, 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 :~50 mg/mL (~95.73 mM)
DMSO :~25 mg/mL (~47.86 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.08 mg/mL (3.98 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (3.98 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 20.8 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: ≥ 2.08 mg/mL (3.98 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 (191.46 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 | 1.9146 mL | 9.5729 mL | 19.1457 mL | |
| 5 mM | 0.3829 mL | 1.9146 mL | 3.8291 mL | |
| 10 mM | 0.1915 mL | 0.9573 mL | 1.9146 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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