Ricolinostat (ACY-1215)

Catalog No.S8001 Synonyms: Rocilinostat

Ricolinostat (ACY-1215) Chemical Structure

Molecular Weight(MW): 433.5

Ricolinostat (ACY-1215) is a selective HDAC6 inhibitor with IC50 of 5 nM in a cell-free assay. It is >10-fold more selective for HDAC6 than HDAC1/2/3 (class I HDACs) with slight activity against HDAC8, minimal activity against HDAC4/5/7/9/11, Sirtuin1, and Sirtuin2. Phase 2.

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Cited by 15 Publications

6 Customer Reviews

  • (a) Representative images from hTERT RPE-1 cells transiently transfected with siControl (siC), treated with vehicle (DMSO), alisertib (MLN8237) or rocilinostat (ACY1215) at the time of serum withdrawal for 48 h. Ciliation monitored by immunofluorescent staining using acetylated α-tubulin (cilia marker) and pericentrin (basal body marker). Nuclei counterstained using DAPI. Highlighted boxes show magnified cilia. Scale bar, 3 μM.

    Oncogene, 2017, 36(24):3450-3463. Ricolinostat (ACY-1215) purchased from Selleck.

  • Immunoblot of MB99-1 cells treated with the indicated compound concentrations for 48 hours. Note that the HDAC6-selective inhibitors affect tubulin acetylation, but not histone H3 acetylation, whereas the pan-HDAC inhibitor TSA increases the acetylation of both.

    Mol Cancer Ther, 2015, 14(3):727-39.. Ricolinostat (ACY-1215) purchased from Selleck.

  • Western blot analysis of PARP, C-caspase-3, total MET and phosphorylated MET (p-MET) in NuDUL-1 and Toledo cells treated as metioned. GAPDH served as a loading control. NC=negative control.

    J Pathol, 2018, 246(2):141-153. Ricolinostat (ACY-1215) purchased from Selleck.

  • Effect of 9 h treatment with bortezomib (Bort, 5 nM) alone or in combination with C1A (2 µM) or ACY-1215 (ACY, 2 µM) on caspase-3/7 activity in OPM-2 cells.

    Br J Cancer, 2018, 119(10):1278-1287. Ricolinostat (ACY-1215) purchased from Selleck.

  • WB analysis of PARP and acetylated-α-tubulin in CRC cells following treatment with ACY-1215 for 24h.

    Mol Cancer Ther, 2018, 17(6):1280-1290. Ricolinostat (ACY-1215) purchased from Selleck.

  • E. The expression levels of TP and TS mRNA were evaluated by qRealTime-PCR in untreated MCF-7 cells and in MCF-7 cells that were untreated or treated with selective HDACi at the indicated concentrations for 24 hours. F. The expression of TP, TS, acetylated H3 and acetylated α-tubulin proteins was determined by western blot in untreated cells and in MCF-7 cells that were treated for 48 hours with different HDACi at the indicated concentrations. α-tubulin was used as the protein loading control

    Oncotarget, 2016, 7(7):7715-31. Ricolinostat (ACY-1215) purchased from Selleck.

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Biological Activity

Description Ricolinostat (ACY-1215) is a selective HDAC6 inhibitor with IC50 of 5 nM in a cell-free assay. It is >10-fold more selective for HDAC6 than HDAC1/2/3 (class I HDACs) with slight activity against HDAC8, minimal activity against HDAC4/5/7/9/11, Sirtuin1, and Sirtuin2. Phase 2.
Features Induced less cytotoxicity in PHA-stimulated PBMCs from 4 healthy donors compared with the pan-HDAC inhibitor SAHA.
Targets
HDAC6 [1]
(Cell-free assay)		 HDAC2 [1]
(Cell-free assay)		 HDAC3 [1]
(Cell-free assay)		 HDAC1 [1]
(Cell-free assay)		 HDAC8 [1]
(Cell-free assay)
4.7 nM 48 nM 51 nM 58 nM 100 nM
In vitro

ACY-1215 is a hydroxamic acid derivative. ACY-1215 is 12-, 10-, and 11-fold less active against HDAC1, HDAC2, and HDAC3 (class I HDACs), respectively. ACY-1215 has minimal activity (IC50 > 1μM) against HDAC4, HDAC5, HDAC7, HDAC9, HDAC11, Sirtuin1, and Sirtuin2, and has slight activity against HDAC8 (IC50 = 0.1μM). The IC50 values for ACY-1215 for T-cell toxicity is 2.5μM. ACY-1215 overcomes tumor cell growth and survival conferred by BMSCs and cytokines in the BM milieu. ACY-1215 in combination with bortezomib induces synergistic anti-MM activity. ACY-1215 induces potent acetylation of α-tubulin at very low doses and triggers acetylation of lysine on histone H3 and histone H4 only at higher doses, confirming its specific inhibitory effect on HDAC6 activity. [1]
Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
RPMI8226 NIixUoZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M{\YXWlEPTB;MUS2PEDDuSB|MUCgcm0> M4ezb|I3PDR|MEe4
A-172 MWHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NXrLNnVtOTEEoH7N MmDQNlQwPDhiaB?= NV\5cW01cW6qaXLpeJMh[2WubDDndo94fGhidHnt[UBl\XCnbnTlcpRtgQ>? NFLzR4kzPjF3MEO0NC=>
U87MG MUTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MoPMNVDDqG6P Mmm1NlQwPDhiaB?= NGTxOnNqdmirYnn0d{Bk\WyuIHfyc5d1cCC2aX3lJIRmeGWwZHXueIx6 M4Xm[lI3OTVyM{Sw
Hbl-1 NXzldW1nT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NUHs[lJOPDhiaB?= MWnJR|UxRTFwNjFOwG0> MVKyOlEyPjJ5MB?=
OCI-Ly10 NIfXPFJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M1LrcVQ5KGh? MXPJR|UxRTBwOTFOwG0> MnuzNlYyOTZ{N{C=
Riva MkXOS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NFXlPXg1QCCq MYnJR|UxRTJwMjFOwG0> MWOyOlEyPjJ5MB?=
Su-DHL2 MlnPS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NF3M[pM1QCCq NHjRcoRKSzVyPUOuN{DPxE1? NF3uT4IzPjFzNkK3NC=>
OCI-Ly1 NX[xUINrT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NE\YTZM1QCCq MojNTWM2OD1{LkSg{txO MlL2NlYyOTZ{N{C=
OCI-Ly7 NXHUVGZUT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MWW0PEBp NICxe3JKSzVyPUGuNkDPxE1? MkO1NlYyOTZ{N{C=
Su-DHL4 NX:4XIxIT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NFzERWc1QCCq MUnJR|UxRTRwNzFOwG0> NEeyNGIzPjFzNkK3NC=>
Su-DHL6 MnjIS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NGP5WGQ1QCCq M{XvbWlEPTB;Mz6yJO69VQ>? MmniNlYyOTZ{N{C=
Hbl-2 M1TzXWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MUW0PEBp MVHJR|UxRTFwOTFOwG0> MXGyOlEyPjJ5MB?=
Jeko-1 NVqwW4FwT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NFi2XnM1QCCq NFjrTHNKSzVyPUGuOUDPxE1? MoHKNlYyOTZ{N{C=
Jvm-2 MlGyS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MnH0OFghcA>? MnfOTWM2OD12LkCg{txO Mle2NlYyOTZ{N{C=
Rec-1  NFfoOZZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NVv1bIhzPDhiaB?= M1HnPGlEPTB;Mj6zJO69VQ>? NILyT4czPjFzNkK3NC=>
CCL-119 MYjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MX20PEBp NVvG[nlsUUN3ME2xMlch|ryP MVyyOlEyPjJ5MB?=
H9 M4fqUGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NIjaZYg1QCCq NVSzTHZNUUN3ME2xMlIh|ryP MoLyNlYyOTZ{N{C=
HH NXrOSGpIT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MYi0PEBp MlT6TWM2OD1{LkWg{txO MVyyOlEyPjJ5MB?=
Sup-T1 Ml3TS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M2XGXVQ5KGh? M37LR2lEPTB;MT62JO69VQ>? M1Lp[|I3OTF4Mkew
MM.1S M1PHfWZ2dmO2aX;uJGF{e2G7 NYjzWZh{OC13zszN MYm2JIg> NYXyToVucW6lcnXhd4V{KGGlZYT5cIF1\WRizsGteJVjfWyrbh?= M1fmblIzOjZ{N{[w
MM.1S MkmySpVv[3Srb36gRZN{[Xl? MV[wMlI2NzIQvF2= MoL6NVghcA>? MkPhbY5kemWjc3XzJIFk\XS7bHH0[YQh|rFvdIXieYxqdg>? NGTSVFYzOjJ4Mke2NC=>
MM.1R NILhdJVHfW6ldHnvckBCe3OjeR?= Mn;ZNE4zPS9zzszN M1n4VFE5KGh? MY\pcoNz\WG|ZYOgZYNmfHmuYYTl[EDPuS22dXL1cIlv MYWyNlI3Ojd4MB?=
RPMI8226  NV25PZp6TnWwY4Tpc44hSXO|YYm= MlLzNE4zPS9zzszN M{XkfFE5KGh? Ml\4bY5kemWjc3XzJIFk\XS7bHH0[YQh|rFvdIXieYxqdg>? NHrTdWszOjJ4Mke2NC=>
MM.1S NVLCb|J6S2WubDDWbYFjcWyrdImgRZN{[Xl? M1r5PFAuQM7:TR?= NFLV[Jg1QCCq NELwNWxl\WO{ZXHz[ZMhVU1vY3XscEB3cWGkaXzpeJkhcW5iYTDkc5NmNWSncHXu[IVvfCCvYX7u[ZI> NXPkbVlJOjJ{NkK3OlA>
OPM1 NUPvNlRoS2WubDDWbYFjcWyrdImgRZN{[Xl? MWiwMVjPxE1? MnWzOFghcA>? M3XtUIRm[3KnYYPld{BOVS2lZXzsJJZq[WKrbHn0fUBqdiCjIHTvd4Uu\GWyZX7k[Y51KG2jbn7ldi=> NV\Hc2dxOjJ{NkK3OlA>
RPMI NXXr[4pzS2WubDDWbYFjcWyrdImgRZN{[Xl? Mlj4NE05|ryP NUnSXZdLPDhiaB?= MWPk[YNz\WG|ZYOgUW0u[2WubDD2bYFjcWyrdImgbY4h[SCmb4PlMYRmeGWwZHXueEBu[W6wZYK= MofhNlIzPjJ5NkC=
MM.1R MXHD[YxtKF[rYXLpcIl1gSCDc4PhfS=> M3\mflAuQM7:TR?= Ml7KOFghcA>? MoXS[IVkemWjc3XzJG1ONWOnbHygeoli[mmuaYT5JIlvKGFiZH;z[U1l\XCnbnTlcpQhdWGwbnXy M{mwR|IzOjZ{N{[w
LR5 MWHD[YxtKF[rYXLpcIl1gSCDc4PhfS=> M2\OSFAuQM7:TR?= M{[yUVQ5KGh? NIHJV3hl\WO{ZXHz[ZMhVU1vY3XscEB3cWGkaXzpeJkhcW5iYTDkc5NmNWSncHXu[IVvfCCvYX7u[ZI> NVLST3JoOjJ{NkK3OlA>
OPM2 NFXOWWNE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MnLDNE05|ryP NUD6Xmh1PDhiaB?= NHm4RXhl\WO{ZXHz[ZMhVU1vY3XscEB3cWGkaXzpeJkhcW5iYTDkc5NmNWSncHXu[IVvfCCvYX7u[ZI> MnT4NlIzPjJ5NkC=

... Click to View More Cell Line Experimental Data
Assay
Methods Test Index PMID
Western blot
Ac-α-tubulin / Ac-Histone H4 ; 

PubMed: 31015208     


Ricolinostat promotes the accumulation of acetylated (Ac) α-tubulin. MM cells were treated with the indicated concentrations of ricolinostat for 24 hours. Protein expression was measured by immunoblotting. 

Survivin / P21 / CDC2 / p53 / p-p53(S392) / Cyclin A2 / Cyclin B1; 

PubMed: 30050135     


Western blot analysis of G2/M phase cell cycle regulatory-proteins for 48 h

Bax / Bim / Bcl2 / Cleaved caspase-3 / Cleaved caspase-9 / Cleaved PARP; 

PubMed: 30050135     


Western blot analysis of apoptosis regulatory-proteins for 48 h

PI3K(p85) / AKT / p-AKT(S473) / PRAS40 / Rag C / mTOR / p-mTOR / ERK / p-ERK; 

PubMed: 30050135     


ACY-1215 treatment inhibited PI3K/AKT/mTOR and ERK signaling protein levels in ESCC EC109 and TE-1 cell lines for 48 h.

Ac-β-catenin(K49) / p-β-catenin / β-catenin; 

PubMed: 25546293     


Time-course immunoblot analysis of human NPCs treated with an HDAC6 inhibitor. Human NPCs were treated with HDAC6 inhibitor ACY-1215 at 5 μM for the times points indicated and the lysates immunoblotted with antibodies against β-catenin, Ac-Lys49-β-catenin, phos-Ser45 and phos-Ser33, Ser37, and Thr41. β-actin is using shown as loading control.

31015208 30050135 25546293
Immunofluorescence
β-tubulin / β-catenin; 

PubMed: 25546293     


Immunofluorescence staining for β-catenin treated with HDAC6 inhibitor in human NPCs. Cells were treated with 5 μM ACY-1215 or DMSO for 18 h and imaged with anti-β-catenin antibody (green), anti-β-tubulin antibody (red), and Hoechst 33342 (blue). Scale bar, 20 μm.  Quantification of β-catenin levels at the membrane represent two independent experiments with three fields of view each at 20× magnification. Error bars indicate standard deviation. ** and **** denote significance at p < 0.01 and p < 0.0001, respectively (unpaired t test).

25546293
Growth inhibition assay
Cell viability; 

PubMed: 31015208     


Ricolinostat decreases MM cell viability. A panel of MM cell lines was treated with the indicated concentrations of ricolinostat for 72 hours. Cell viability was measured by MTT assay.

31015208
In vivo ACY-1215 in combination with bortezomib triggered more significant anti-MM activity than either agent alone in suppressing tumor growth and prolonging survival in both plasmacytoma model and disseminated MM model without significant adverse effects. ACY-1215 is readily absorbed by tumor tissue. Moreover, the drug does not accumulate in tumor tissue, as evidenced by the parallel decline of acetylated α-tubulin in blood cells and tumor tissue by 24 hours after dose. [1]

Protocol

Kinase Assay:

[1]
+ Expand

HDAC enzymatic assays:

ACY-1215 is dissolved and subsequently diluted 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] to 6-fold the final concentration. HDAC enzymes are diluted to 1.5-fold of the final concentration in assay buffer and pre-incubated with ACY-1215 for 10 minutes before the addition of the substrate. The amount of FTS (HDAC1, HDAC2, HDAC3, and HDAC6) or MAZ-1675 (HDAC4, HDAC5, HDAC7, HDAC8, and HDAC9) used for each enzyme is equal to the Michaelis constant (Km), as determined by a titration curve. FTS or MAZ-1675 is diluted in assay buffer to 6-fold the final concentration with 0.3μM sequencing grade trypsin. The substrate/trypsin mix is added to the enzyme/compound mix and the plate is shaken for 60 seconds and then placed into a SpectraMax M5 microtiter plate reader. The enzymatic reaction is monitored for release of 7-amino-4-methoxy-coumarin over 30 minutes, after deacetylation of the lysine side chain in the peptide substrate, and the linear rate of the reaction is calculated.
Cell Research:

[1]
+ Expand
  • Cell lines: MM cell lines, patient MM cells, and PBMCs
  • Concentrations: ~8 μM
  • Incubation Time: 48 hours
  • Method:

    PBMCs from healthy donors are isolated and stimulated with 2.5 μg/mL of phytohemagglutinin (PHA) for 48 hours in the presence of increasing concentrations of ACY-1215. DNA synthesis is measured by tritiated thymidine uptake. CD4+T cells are purified from human blood with the Rosette Sep negative-selection kit. Cells are stimulated by CD3/CD28 Dynabeads for 7 days in the presence of compounds. Cell viability is assessed using alamarBlue. MM cells (2-3 × 104cells/well) are incubated in 96-well culture plates with medium and 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) for 24 hours at 37°C, and tritiated thymidine incorporation is measured.


    (Only for Reference)
Animal Research:

[1]
+ Expand
  • Animal Models: MM xenograft SCID mouse model
  • Formulation: 10% DMSO in 5% dextrose in water
  • Dosages: 50 mg/kg
  • Administration: ip
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 86 mg/mL (198.38 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+ddH2O
For best results, use promptly after mixing. 		5mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 433.5
Formula

C24H27N5O3
CAS No. 1316214-52-4
Storage powder
in solvent
Synonyms Rocilinostat

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

  • Question 1:

    What would you suggest to obtain a clear solution?

  • Answer:

    S8001 ACY-1215 can be dissolved in 2% DMSO/30% PEG 300/ddH2O at 5 mg/ml clearly while it in 1% DMSO/30% polyethylene glycol/1% Tween 80 at 30 mg/ml is a suspension for oral administration. Please note that the precipitation will go out from the clear solution after stayed for about half an hour, So it is recommended to prepare the solution just before use.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID