ACY-775

Alias: ACY 775; ACY775; ACY-775

Cat No.:V3772 Purity: ≥98%

COA Product Data Instructions for use SDS

ACY-775, anovel pyrimidine hydroxyl amide small molecule, is a brain bioavailable, highly potent and isoform-selective inhibitor of HDAC6 with IC50of 7.5 nM.

ACY-775 Chemical Structure CAS No.: 1375466-18-4
Product category: HDAC

This product is for research use only, not for human use. We do not sell to patients.

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Citations by Top Journals: Nature, Cell, Science, etc.

Top Publications Citing lnvivochem Products

Nature 2021, 597(7874):119-125.

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

Nature 597, 119–125 (2021)

Cell Stem Cell 2020,26(6):845-861.e12.

Science Trans Med 2023,15(701):eadd7872.

STTT 2023,8(1):91.

Cell Reports 2023,42(4):112313

Science Trans Med 2023, 15: eadd7872.

Cancer Cell 2021,39(4):529-547.e7.

Cancer Discov 2022,12(4):1106-1127.

Immunity 2023,56(3):620-634.e11.

J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

Biomaterial 2023 Sep16:302:122333.

Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

ACY-775, a novel pyrimidine hydroxyl amide small molecule, is a brain bioavailable, highly potent and isoform-selective inhibitor of HDAC6 with IC50 of 7.5 nM. ACY-738 and ACY-775 exhibit a selectivity of 60–1500 times greater than class I HDACs while inhibiting HDAC6 with low nanomolar potency. When it comes to stimulating mouse exploratory behaviors in unfamiliar but novel environments, ACY-738 and ACY-775 cause dramatic increases in α-tubulin acetylation in the brain, unlike tubastatin A, a reference HDAC6 inhibitor with similar potency and peripheral activity but more limited brain bioavailability. Remarkably, ACY-738 and ACY-775 exhibit antidepressant-like effects in the tail suspension test and social defeat paradigm, similar to other HDAC inhibitors like SAHA and MS-275, despite having no discernible effect on histone acetylation. The central inhibition of HDAC6 is directly responsible for the effects of ACY-738 and ACY-775, as these effects are completely abolished in mice that have a loss of function of HDAC6 that is specific to the neurons. Moreover, a behaviorally inactive dosage of ACY-738 combined with a subactive dose of the selective serotonin reuptake inhibitor citalopram significantly enhances the anti-immobility effect. These findings support the viability of this HDAC isoform as a possible target for the development of antidepressants and validate novel isoform-selective probes for in vivo pharmacological investigations of HDAC6 in the CNS.

Biological Activity I Assay Protocols (From Reference)

Targets

HDAC6 (IC50 = 7.5 nM); HDAC1 (IC50 = 2123 nM); HDAC2 (IC50 = 2570 nM); HDAC3 (IC50 = 11223 nM)

ln Vitro

In cells treated with the vehicle, histone 3 is obviously acetylated, but α-tubulin is primarily seen in its deacetylated state. ACY-775 treatment results in a definite increase in α-tubulin acetylation, but not in histone acetylation. Through the use of immunofluorescence, α-tubulin acetylation can be seen, and the intensity of this signal in the neuronal neurites can be measured and compared to its length. The DRG neurons treated with vehicles already have acetylated α-tubulin. Acetylated α-tubulin significantly increases in signal intensity after treatment with ACY-775. When comparing the neurites to DRG neurons treated with a vehicle, a notable increase in both the motility and total number of mitochondria was noted. Compared to cells treated with a vehicle, DRG neurons treated with ACY-775 exhibit a noticeably higher number of retrogradely transport mitochondria[1].

ln Vivo

ACY-738, ACY-775, and tubastatin A biodistribution profiles are investigated following acute dosing at 5 or 50 mg/kg over a 2-hour period. The plasma levels of ACY-738 and ACY-775 at t=30 min following an acute 50 mg/kg injection are 515 ng/mL (1.9 μM) and 1359 ng/mL (4.1 μM), respectively. It is rapidly eliminated from plasma; after two hours, the concentration falls below 10 ng/mL, with a plasmatic half-life of 12 minutes. However, ratios greater than one result from areas under concentration time curves for brain and plasm computed over a 2-hour period for both ACY-738 and ACY-775. In wild-type mice, significant increases in α-tubulin acetylation are observed in all tested brain regions when ACY-738 (5 mg/kg) or ACY-775 (50 mg/kg) are administered repeatedly at 24 h, 4 h, and 30 min before killing[2].

Cell Assay

The line of immortalized rat raphe neuronal precursors, known as undifferentiated RN46A-B14 cells, is cultured. For four hours, they are given either 2.5 μM ACY-738, ACY-775, tubastatin A, 0.6 μM TSA, or 0.1% DMSO as a vehicle. Samples are prepared with a histone extraction kit, and protein assay is used to quantify the results.

Animal Protocol

5, 50 mg/kg; i.p.

In the TST, mice are tested for immobility. A combination of the previous, or vehicle, mice are attached to the test rig at 30 minutes or 2 hours after intraperitoneal injection of ACY-738 (5, 50 mg/kg), ACY-775 (5, 50 mg/kg), and citalopram (0.5, 2, 20 mg/kg). The amount of time the mice remain immobile for 6 minutes is recorded. Mice are injected with ACY-738 or ACY-775 at 5, 10, or 50 mg/kg or a vehicle, and then given free rein to explore in an open-field environment. Recorded activity exists [2].

References

[1]. Development of Improved HDAC6 Inhibitors as Pharmacological Therapy for Axonal Charcot-Marie-Tooth Disease. Neurotherapeutics. 2017 Apr; 14(2): 417-428.

[2]. Antidepressant-Like Properties of Novel HDAC6-Selective Inhibitors with Improved Brain Bioavailability. Neuropsychopharmacology. 2014 Jan; 39(2): 389-400.

[3]. Target deconvolution of HDAC pharmacopoeia reveals MBLAC2 as common off-target. Nat Chem Biol. 2022 Apr 28.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula

C17H19FN4O2

Molecular Weight

330.36

Exact Mass

330.15

Elemental Analysis

C, 61.81; H, 5.80; F, 5.75; N, 16.96; O, 9.69

CAS #

1375466-18-4

Related CAS #

1375466-18-4

PubChem CID

57382288

Appearance

White to off-white solid powder

Density

1.1±0.1 g/cm3

Boiling Point

589.7±45.0 °C at 760 mmHg

Flash Point

310.4±28.7 °C

Vapour Pressure

0.0±1.7 mmHg at 25°C

Index of Refraction

1.566

LogP

1.54

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

423

Defined Atom Stereocenter Count

SMILES

FC1=CC=CC(=C1)C1(CCCCC1)NC1N=CC(C(NO)=O)=CN=1

InChi Key

IYBURCQQEUNLDL-UHFFFAOYSA-N

InChi Code

InChI=1S/C17H19FN4O2/c18-14-6-4-5-13(9-14)17(7-2-1-3-8-17)21-16-19-10-12(11-20-16)15(23)22-24/h4-6,9-11,24H,1-3,7-8H2,(H,22,23)(H,19,20,21)

Chemical Name

2-[[1-(3-fluorophenyl)cyclohexyl]amino]-N-hydroxypyrimidine-5-carboxamide

Synonyms

ACY 775; ACY775; ACY-775

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)

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL

Water: N/A

Ethanol: N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.57 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 (7.57 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.57 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	3.0270 mL	15.1350 mL	30.2700 mL
	5 mM	0.6054 mL	3.0270 mL	6.0540 mL
	10 mM	0.3027 mL	1.5135 mL	3.0270 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.

Calculator

Mass

Concentration

Volume

Molecular Weight*

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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See Example

An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?

Enter 350.26 in the Molecular Weight (MW) box
Enter 10 in the Concentration box and choose the correct unit (mM)
Enter 5 in the Volume box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Concentration (Start)

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Concentration (End)

Volume (End)

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:

Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
Enter 25 into the Concentration (End) box and select the correct unit (mM)
Enter 25 into the Volume (End) box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

Molecular formula

Total molecular weight

g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4 c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Volume (to add to vial)

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Desired Reconstitution Concentration

Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
Click the “Calculate” button
The answer appears in the Volume (to add to vial) box

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)

Dosage mg/kg

Average weight of animals g

Dosing volume per animal μL

Number of animals

Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)

% DMSO

% Tween 80

% ddH₂O

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.

Note： (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.

Biological Data

Selectivity, potency, and pharmacokinetic properties of HDAC6 inhibitors used in this study.Neuropsychopharmacology.2014 Jan;39(2):389-400.
Effects of HDAC6 inhibitors onα-tubulin acetylation at lysine 40 (K40) and histone H3 acetylation at lysine 9 (H3K9).Neuropsychopharmacology.2014 Jan;39(2):389-400.
HDAC6 inhibitors ACY-738 and ACY-775 have antidepressant-like properties.Neuropsychopharmacology.2014 Jan;39(2):389-400.
ACY-738 and ACY-775 are potent and selective histone deacetylase 6 inhibitors.Neurotherapeutics.2017 Apr;14(2):417-428.

Histone deacetylase 6 (HDAC6) inhibition using ACY-738 and ACY-775 rescued the mitochondrial defects in dorsal root ganglion (DRG) neurons cultured from symptomatic HSPB1S135Fmice.Neurotherapeutics.2017 Apr;14(2):417-428.

Histone deacetylase 6 (HDAC6) inhibition using ACY-738, ACY-775, or ACY-1215 reversed the axonal deficits in motor and sensory nerves and induced reinnervation of neuromuscular junction.Neurotherapeutics.2017 Apr;14(2):417-428.