 Chemical Structure.png)
BML-210 (also known as BML210; CAY-10433) is novel and potent HDAC inhibitor (IC50 value: 5 μM) with anticancer activity. BML-210 induces growth inhibition and apoptosis and regulates HDAC and DAPC complex expression levels in cervical cancer cells. BML-210 can inhibit cell growth and induce apoptosis in cervical cancer cells, what correlates with down-regulation of HDAC class I and II and changes in the DAPC expression levels. Cell cycle analysis indicated that HeLa cell treatment with 20 and 30 μM concentration of BML-210 increased the proportion of cells in G0/G1 phase, and caused accumulation in subG1, indicating that the cells are undergoing apoptosis.
Physicochemical Properties
| Molecular Formula | C20H25N3O2 |
| Molecular Weight | 339.4314 |
| Exact Mass | 339.194 |
| Elemental Analysis | C, 70.77; H, 7.42; N, 12.38; O, 9.43 |
| CAS # | 537034-17-6 |
| PubChem CID | 9543540 |
| Appearance | White to off-white solid powder |
| Density | 1.2±0.1 g/cm3 |
| Boiling Point | 632.5±40.0 °C at 760 mmHg |
| Flash Point | 336.3±27.3 °C |
| Vapour Pressure | 0.0±1.9 mmHg at 25°C |
| Index of Refraction | 1.633 |
| LogP | 2.58 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 3 |
| Rotatable Bond Count | 9 |
| Heavy Atom Count | 25 |
| Complexity | 408 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C(N([H])C1C([H])=C([H])C([H])=C([H])C=1[H])=O)N([H])C1=C([H])C([H])=C([H])C([H])=C1N([H])[H] |
| InChi Key | RFLHBLWLFUFFDZ-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C20H25N3O2/c21-17-12-8-9-13-18(17)23-20(25)15-7-2-1-6-14-19(24)22-16-10-4-3-5-11-16/h3-5,8-13H,1-2,6-7,14-15,21H2,(H,22,24)(H,23,25) |
| Chemical Name | N-(2-aminophenyl)-N'-phenyl-octanediamide |
| Synonyms | BML-210; CAY10433; BML 210; CAY-10433; BML210; BML-210; 537034-17-6; N1-(2-aminophenyl)-N8-phenyloctanediamide; BML-210(CAY10433); N-(2-aminophenyl)-N'-phenyl-octanediamide; CHEBI:61077; Octanediamide, N-(2-aminophenyl)-N'-phenyl-; CAY 10433. |
| 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) |
Biological Activity
| Targets | HDAC4; HDAC4-VP16-driven reporter |
| ln Vitro | NB4 cells are inhibited in their growth and proliferation by BML-210 (10, 20 μM; 24, 48 hours) [2]. The amount of S phase NB4 cells was reduced by BML-210 (10, 20 μM; 24, 48 hours), and at 20 μM, G0 BML-210 (10, 20 μM; 24, 48 hours) had a cytotoxic effect on NB4 cells. A dose of 10 μM is sufficient for BML-210 to cause cell death [2]. In NBT4 cells, BML-210 (10, 20 μM; 24, 48 hours) suppresses HDAC expression and activity [2]. HDAC4-VP16 expression is not downregulated by BML-210[1].Cell lines. NB4 cells Concentration: 10, 20 μM Incubation period: 24, 48 hours Outcomes: Reduced NB4 cell growth and proliferation in a time- and dose-dependent manner. |
| ln Vivo |
BML-210 (20 mg/kg; intraperitoneal injection; three times weekly for two weeks) markedly reduced the amount of body weight and tumor growth. In immunodeficient nude mice, BML-210 has no influence on tumor growth or body weight (Nu/J).
In this study, researchers show that the epigenetic inhibitors GSK-LSD1, CUDC-101 and BML-210, identified by the screen, display antitumour activities in orthotopic mammary tumours in mice, that they upregulate antigen presentation mediated by the major histocompatibility complex class I on breast tumour cells and that treatment with BML-210 substantially sensitized breast tumours to the inhibitor of the checkpoint programmed death-1. Standardized measurements of tumour-cell killing activity facilitated by tumour-organoid-T-cell screens may help with the identification of candidate immunotherapeutics for a range of cancers[3]. |
| Enzyme Assay |
HDAC Activity Analysis[2] Proteins were isolated using ProteoJET™ Mammalian Cell Lysis Reagent (Fermentas, Vilnius, Lithuania) according to the manufacturer's instructions. HDAC activity analysis was determined using EpiQuik™ HDAC Activity/Inhibition Assay Kit according to the manufacturer’s instructions. Absorbance was measured using Tecan-Control, Infinite 200 microplate reader at 450 nm. Activity (ng/h/mL) was calculated using formula.[2] Drug treatment and luciferase assay[1] For the two-hybrid assay, cells were treated with 10 µM drug overnight unless indicated otherwise. The amount of DMSO was kept below 0.2% V/V. A luciferase assay was performed according to the manufacturer’s protocol. The luciferase response was normalized against the Renilla Luciferase as an internal control. The data are presented as a mean ± SD (n = 2) of normalized HDAC4:MEF2 luciferase response against the normalized response values for GAL4-VP16 for each condition to correct for non-specific inhibition of the luciferase signal. |
| Cell Assay |
Cell proliferation assay[2] Cell Types: NB4 cells Tested Concentrations: 10, 20 μM Incubation Duration: 24, 48 hrs (hours) Experimental Results: Inhibited cell proliferation and inhibited the growth of NB4 cells in a dose- and time-dependent manner. Cell cycle analysis [2] Cell Types: NB4 cells Tested Concentrations: 10, 20 μM Incubation Duration: 24, 48 hrs (hours) Experimental Results: The proportion of S phase of NB4 cells diminished and the proportion of G0/G1 phase increased. 10 μM increased the G0/G1 phase by up to 70% at 24 and 48 hrs (hours). Cytotoxicity assay [2] Cell Types: NB4 cells Tested Concentrations: 10, 20 μM Incubation Duration: 24, 48 hrs (hours) Experimental Results: Cytotoxic effects on NB4 cells were dose- and time-dependent. Apoptosis analysis [2] Cell Types: NB4 Cell Tested Concentrations: 10, 20 μM Incubation Duration: 24, 48 hrs (hours) Experimental Results: 10 μM dose induced apoptosis. Western Blot Analysis[2] Cell Types: NB4 Cell Tested Concentrations: 10, 20 μM Incubation Duration: 24, 48 hrs (hours) Experimental Results: After 48 hrs (hours) of treatment with 10 μM dose, HDAC1 gene expression was inhibited by up to 36% Aft |
| Animal Protocol |
Animal/Disease Models: Female C57BL/6 mice, mouse breast cancer EO771 cells [3] Doses: 20 mg/kg Route of Administration: IP ;[3]. Three times a week for two weeks. Experimental Results: Significant inhibition of tumor growth and weight. |
| References |
[1]. Inhibition of the function of class IIa HDACs by blocking their interaction with MEF2. Nucleic Acids Res. 2012 Jul; 40(12): 5378–5388. [2]. The Histone Deacetylase Inhibitor BML-210 Influences Gene and Protein Expression in Human Promyelocytic Leukemia NB4 Cells via Epigenetic Reprogramming. Int J Mol Sci. 2015 Aug; 16(8): 18252–18269. [3]. An organoid-based screen for epigenetic inhibitors that stimulate antigen presentation and potentiate T-cell-mediated cytotoxicity. Nat Biomed Eng. 2021 Nov;5(11):1320-1335. |
| Additional Infomation | BML-210 is a dicarboxylic acid diamide comprising suberic (octanedioic) acid coupled to aniline and 1,2-diaminobenzene. It has a role as an EC 3.5.1.98 (histone deacetylase) inhibitor and an antineoplastic agent. It is functionally related to a suberic acid. |
Solubility Data
| Solubility (In Vitro) | DMSO : ≥ 30 mg/mL (~88.38 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.37 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9461 mL | 14.7306 mL | 29.4612 mL | |
| 5 mM | 0.5892 mL | 2.9461 mL | 5.8922 mL | |
| 10 mM | 0.2946 mL | 1.4731 mL | 2.9461 mL |