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Rocilinostat (formerly Ricolinostat; ACY-1215), an investigational drug, is a potent, orally bioavailable and selective HDAC6 inhibitor with potential anticancer activity. In a cell-free experiment, it inhibits HDAC6 with an IC50 of 5 nM, exhibits over ten-fold selectivity for HDAC6 over HDAC1/2/3, and exhibits negligible or no activity against HDAC8
Physicochemical Properties
| Molecular Formula | C24H27N5O3 | |
| Molecular Weight | 433.5 | |
| Exact Mass | 433.211 | |
| Elemental Analysis | C, 66.50; H, 6.28; N, 16.16; O, 11.07 | |
| CAS # | 1316214-52-4 | |
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| PubChem CID | 53340666 | |
| Appearance | White to pink pale peach fluffy powder | |
| Density | 1.2±0.1 g/cm3 | |
| Index of Refraction | 1.620 | |
| LogP | 1.41 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 6 | |
| Rotatable Bond Count | 11 | |
| Heavy Atom Count | 32 | |
| Complexity | 538 | |
| 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])N([H])C(C1C([H])=NC(=NC=1[H])N(C1C([H])=C([H])C([H])=C([H])C=1[H])C1C([H])=C([H])C([H])=C([H])C=1[H])=O)N([H])O[H] |
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| InChi Key | QGZYDVAGYRLSKP-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C24H27N5O3/c30-22(28-32)15-9-1-2-10-16-25-23(31)19-17-26-24(27-18-19)29(20-11-5-3-6-12-20)21-13-7-4-8-14-21/h3-8,11-14,17-18,32H,1-2,9-10,15-16H2,(H,25,31)(H,28,30) | |
| Chemical Name | N-[7-(hydroxyamino)-7-oxoheptyl]-2-(N-phenylanilino)pyrimidine-5-carboxamide | |
| Synonyms | Ricolinostat; Rocilinostat; ACY 1215; ACY1215; ACY-1215 | |
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
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| 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 | HDAC6 ( IC50 = 4.7 nM ); HDAC2 ( IC50 = 48 nM ); HDAC3 ( IC50 = 51 nM ); HDAC1 ( IC50 = 58 nM ); HDAC8 ( IC50 = 100 nM ); HDAC7 ( IC50 = 1400 nM ); HDAC5 ( IC50 = 5000 nM ); HDAC4 ( IC50 = 7000 nM ) | |
| ln Vitro |
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| ln Vivo |
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| Enzyme Assay | ACY-1215 is dissolves and is then diluted to six times the final concentration in assay buffer (50 mM HEPES, pH 7.4, 100 mM KCl, 0.001% Tween-20, 0.05% BSA, and 20 μM tris(2-carboxyethyl)phosphine). HDAC enzymes are pre-incubated with ACY-1215 for 10 minutes prior to the addition of the substrate, and they are diluted to 1.5 times the final concentration in assay buffer. The Michaelis constant (Km), as ascertained by a titration curve, is equivalent to the quantity of FTS (HDAC1, HDAC2, HDAC3, and HDAC6) or MAZ-1675 (HDAC4, HDAC5, HDAC7, HDAC8, and HDAC9) used for each enzyme. FTS or MAZ-1675 is diluted with 0.3μM sequencing grade trypsin in assay buffer to six times the final concentration. The plate is put into a SpectraMax M5 microtiter plate reader after the substrate/trypsin mix has been added to the enzyme/compound mix and shaken for 60 seconds. Following the peptide substrate's lysine side chain's deacetylation, the enzymatic reaction is watched for the release of 7-amino-4-methoxy-coumarin over a 30-minute period. The reaction's linear rate is then computed. | |
| Cell Assay | Peripheral blood mononuclear cells (PBMCs) with immunoblots from healthy donors are isolated and incubated with increasing doses of ACY-1215 for 48 hours while 2.5 μg/mL phytohemagglutinin (PHA) is present. DNA synthesis is measured using thymidine uptake that has been tritium-tagged. Human blood is used to isolate CD4+T cells using the Rosette Sep negative-selection kit. Cells are stimulated by CD3/CD28 Dynabeads for seven days while the compounds are present. AlamarBlue is a tool for viability analysis of cells. 96-well culture plates containing medium, different concentrations of ACY-1215, bortezomib, and/or recombinant IL-6 (10 ng/mL) or insulin-like growth factor-1 (IGF-1; 50 ng/mL) are used to cultivate MM cells (2-4 × 104 cells/well) for a full day at 37°C. We then verify that tritiated thymidine has been incorporated. | |
| Animal Protocol |
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| References |
[1]. Preclinical activity, pharmacodynamic, and pharmacokinetic properties of a selective HDAC6 inhibitor, ACY-1215, in combination with PS-341 in multiple myeloma. Blood. 2012 Mar 15;119(11):2579-89. |
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| Additional Infomation |
N-[7-(hydroxyamino)-7-oxoheptyl]-2-(N-phenylanilino)-5-pyrimidinecarboxamide is a pyrimidinecarboxylic acid. Ricolinostat is under investigation for the treatment of Breast Carcinoma and Metastatic Breast Cancer. Ricolinostat is an orally bioavailable, specific inhibitor of histone deacetylase 6 (HDAC6) with potential antineoplastic activity. Ricolinostat selectively targets and binds to HDAC6, thereby disrupting the Hsp90 protein chaperone system through hyperacetylation of Hsp90 and preventing the subsequent aggresomal protein degradation. This leads to an accumulation of unfolded and misfolded ubiquitinated proteins and may eventually induce cancer cell apoptosis, and inhibition of cancer cell growth. HDAC6, a class II HDAC deacetylase located in the cytoplasm, appears to play a key role in the formation and activation of the aggresomes needed for degradation of misfolded proteins. Compared to non-selective HDAC inhibitor, ACY-1215 is able to reduce the toxic effects on normal, healthy cells. |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.77 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 (5.77 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 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.77 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. Solubility in Formulation 4: 2% DMSO+30% PEG 300+ddH2O: 5mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3068 mL | 11.5340 mL | 23.0681 mL | |
| 5 mM | 0.4614 mL | 2.3068 mL | 4.6136 mL | |
| 10 mM | 0.2307 mL | 1.1534 mL | 2.3068 mL |