| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Targets |
CARM1 (coactivator-associated arginine methyltransferase 1, also known as PRMT4). CARM1 is a type I protein arginine methyltransferase (PRMT) that catalyzes the formation of asymmetric dimethylarginine (ADMA) on histone and non-histone protein substrates. It serves as a transcriptional coactivator for nuclear receptors (e.g., androgen receptor, estrogen receptor) and other transcription factors. CARM1-IN-1 hydrochloride selectively inhibits CARM1 methyltransferase activity, reducing methylation-dependent gene expression and affecting cancer cell proliferation, differentiation, and epigenetic regulation.
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| ln Vitro |
In cell-free biochemical assays, CARM1-IN-1 hydrochloride directly inhibits CARM1 methyltransferase activity with an IC50 of 8.6 uM. It shows very low activity against other methyltransferases, with IC50 > 600 uM for PRMT1 and > 667 uM for SET7. The compound displays high selectivity for CARM1 over a panel of different PRMTs (protein arginine methyltransferases) and HKMTs (histone lysine methyltransferases). In methyltransferase assays using various substrates (PABP1, CA150, SmB, H3), CARM1-IN-1 hydrochloride inhibits the methylation of all tested substrates. The inhibition is competitive with respect to the methyl donor SAM but non-competitive with respect to the peptide substrate in some assays.
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| ln Vivo |
CARM1-IN-1 hydrochloride inhibits CARM1-mediated methylation in cells. In human LNCaP prostate cancer cells, the compound (8 uM; 42-44 hours) shows a significant dose-dependent reduction of PSA (prostate-specific antigen) promoter activity, as measured by luciferase reporter assays. PSA is an androgen-responsive gene whose transcription is coactivated by CARM1. CARM1-IN-1 hydrochloride inhibits PSA promoter activity without affecting cell viability at these concentrations. The compound reduces methylation levels of CARM1 substrates, such as PABP1 and histone H3 (at Arg17), in cells. It does not significantly affect global PRMT1-mediated methylation. No significant cytotoxicity is observed at concentrations up to 25 uM in LNCaP cells.
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| Enzyme Assay |
CARM1 enzymatic activity is measured using a radiometric methyltransferase assay or a luminescent-based assay format. Recombinant CARM1 enzyme (full-length or catalytic domain) is incubated with a biotinylated peptide substrate (e.g., histone H3 1-25 or PABP1-derived peptide), S-adenosyl-[3H]-methionine ([3H]-SAM), and test compound (0.1-200 uM) in assay buffer (Tris-HCl, pH 8.0, NaCl, MgCl2, DTT). After 30-60 minutes at room temperature, the reaction is stopped, and the reaction mixture is transferred to streptavidin-coated scintillation proximity assay (SPA) plates or filter plates. Incorporated radioactivity is counted on a scintillation counter. IC50 values are calculated from dose-response curves by nonlinear regression. Selectivity is assessed by testing against PRMT1, PRMT3, PRMT5, PRMT6, PRMT8, SET7, and other methyltransferases under similar conditions.
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| Cell Assay |
LNCaP human prostate cancer cells (or other CARM1-dependent cell lines) are cultured in RPMI 1640 medium with 10% FBS. For PSA promoter assays, cells are transiently transfected with PSA-luciferase reporter plasmid and an AR expression vector in charcoal-stripped serum-containing medium for 24 hours. Cells are then treated with CARM1-IN-1 hydrochloride (0-25 uM) in the presence of the androgen R1881 (0.1-1 nM) or DHT for 24-48 hours. Luciferase activity is measured using a luciferase assay system, and values are normalized to protein concentration or a constitutively expressed control reporter (e.g., Renilla). For methylation assays, cells are treated with CARM1-IN-1 hydrochloride (1-25 uM) for 24-48 hours, then lysed. Methylation of CARM1 substrates (e.g., PABP1, histone H3) is detected by immunoprecipitation followed by Western blot using anti-methylarginine antibodies or a specific anti-H3R17me2a antibody. Cell viability is assessed using MTT or CellTiter-Glo assays to ensure that observed effects are not due to cytotoxicity.
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| Animal Protocol |
In vivo studies for CARM1-IN-1 hydrochloride are limited. The compound has been primarily used as a biochemical and cell-based tool. For in vivo research, solubility and formulation studies have been conducted. The compound is soluble in DMSO at ≥ 38 mg/mL (64.22 mM) but has poor solubility in water (<0.1 mg/mL insoluble). For in vivo studies, a standard formulation involves sequential addition of DMSO master stock, then PEG300, then Tween 80, then ddH2O or corn oil. A typical formulation for injection uses 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline, achieving ≥ 2.5 mg/mL (4.22 mM) with a clear solution. Doses ranging from 10-50 mg/kg administered intraperitoneally or orally could be used for proof-of-concept studies, but published in vivo efficacy data are not available.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for CARM1-IN-1 hydrochloride are not extensively reported. The compound's hydrochloride salt form improves water solubility compared to the free base, which may enhance absorption. Based on its molecular weight (591.72) and lipophilicity, the compound likely has moderate oral bioavailability. Standard PK parameters (e.g., half-life, Cmax, AUC) would need to be determined in preclinical species for in vivo pharmacology studies. For research use, the compound is typically stored at -20degC in powder form and is stable for at least one year under proper storage conditions. The free acid form (not HCl salt) may have different solubility and stability properties. In vitro, the compound is stable in DMSO solution.
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| Toxicity/Toxicokinetics |
A dedicated toxicology profile for CARM1-IN-1 hydrochloride has not been published. In cell-based assays, CARM1-IN-1 hydrochloride shows no significant cytotoxicity up to 25 uM in LNCaP cells. At higher concentrations (>50 uM), some reduction in cell viability may occur. For in vivo studies, the compound is expected to be reasonably well-tolerated at doses up to 50 mg/kg based on typical small molecule inhibitor safety profiles. As a methyltransferase inhibitor, there may be potential for effects on gene expression that could be associated with long-term toxicity. Standard laboratory safety practices should be followed. The compound has not been evaluated in clinical trials and is not approved for human use.
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| References | |
| Additional Infomation |
CARM1-IN-1 hydrochloride is a research-grade chemical tool for studying CARM1 biology. The compound belongs to the 3,5-bis(bromohydroxybenzylidene)piperidin-4-one chemotype. It was originally described by Cheng et al. and is sometimes referred to as compound 7g. The hydrochloride salt is intended to improve solubility and stability compared to the free base. CARM1 is an emerging target in cancer research, particularly in prostate cancer, breast cancer, and other hormone-dependent malignancies. CARM1-IN-1 hydrochloride is not approved for clinical use and is intended for laboratory research only. In solution, the compound is somewhat unstable; it is recommended to prepare fresh stock solutions immediately before use. The product is for research use only, not for human use. Its molecular formula is C26H22Br2ClNO3·HCl (the free base is C26H22Br2ClNO3). The CAS number for the hydrochloride salt is 2070018-31-2.
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| Molecular Formula |
C26H22BR2CLNO3
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| Molecular Weight |
591.718784809113
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| Exact Mass |
590.963
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| CAS # |
2070018-31-2
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| Related CAS # |
CARM1-IN-1;1020399-49-8
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| PubChem CID |
92044367
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
33
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| Complexity |
670
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| Defined Atom Stereocenter Count |
0
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| SMILES |
BrC1=C(C=CC(=C1)/C=C1/C(/C(=C/C2C=CC(=C(C=2)Br)O)/CN(CC2C=CC=CC=2)C/1)=O)O.Cl
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| InChi Key |
QLPNYHLFFVKOAN-CUBOLJEZSA-N
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| InChi Code |
InChI=1S/C26H21Br2NO3.ClH/c27-22-12-18(6-8-24(22)30)10-20-15-29(14-17-4-2-1-3-5-17)16-21(26(20)32)11-19-7-9-25(31)23(28)13-19;/h1-13,30-31H,14-16H2;1H/b20-10+,21-11+;
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| Chemical Name |
(3E,5E)-1-benzyl-3,5-bis[(3-bromo-4-hydroxyphenyl)methylidene]piperidin-4-one;hydrochloride
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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 is not stable in solution, please use freshly prepared working solution for optimal results. (2). 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) |
DMSO : ~250 mg/mL (~422.50 mM)
H2O : < 0.1 mg/mL |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.22 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 (4.22 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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: 2.08 mg/mL (3.52 mM) in 10% DMSO + 90% Corn Oil (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 | 1.6900 mL | 8.4499 mL | 16.8999 mL | |
| 5 mM | 0.3380 mL | 1.6900 mL | 3.3800 mL | |
| 10 mM | 0.1690 mL | 0.8450 mL | 1.6900 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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