| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg | |||
| Other Sizes |
| Targets |
CHDI-390576 has an affinity (Kd) of 80 nM for the immobilised HDAC4 catalytic domain [1]. CHDI-390576 inhibits class I HDAC (1, 3, 8) and class IIb HDAC6 subtypes at IC50s of 39.7 μM, 25.8 μM, 9.1 μM, and 6.2 μM, respectively [1].
|
|---|---|
| ln Vitro |
CHDI-390576 has an affinity (Kd) of 80 nM for the immobilised HDAC4 catalytic domain [1]. CHDI-390576 inhibits class I HDAC (1, 3, 8) and class IIb HDAC6 subtypes at IC50s of 39.7 μM, 25.8 μM, 9.1 μM, and 6.2 μM, respectively [1].
CHDI-390576 inhibited Class IIa HDAC activity in a cellular assay using Jurkat cells with a Boc-Lys(TFA) substrate, showing an IC₅₀ of 69 nM. It showed no observable activity (IC₅₀ > 50 µM) against Class I/IIb HDACs in a parallel cellular assay using Boc-Lys(Ac) as the exogenous substrate. [1] In A549 cells, CHDI-390576 did not show activity against the canonical Class I substrate histone H4K12 acetylation (IC₅₀ > 30 µM) and showed weak activity against the Class IIb substrate tubulin acetylation with an IC₅₀ of 17 µM. [1] The binding affinity (Kd) and kinetics to the catalytic domain of HDAC4 were determined by surface plasmon resonance (SPR), with the Kd value in good agreement with the biochemical IC₅₀. The dissociation half-life of binding to HDAC4 was approximately 3 minutes. [1] At a test concentration of 10 µM, CHDI-390576 exhibited a clean off-target profile, showing <25% displacement of selective radioligands against an extensive panel of CNS and peripheral receptors, ion channels, enzymes, and transporters. [1] CHDI-390576 was stable in mouse and human plasma and blood, and in simulated gastric fluid. [1] |
| ln Vivo |
In a tolerability study in R6/2 transgenic mice (a model of Huntington's disease) and their wild-type littermates, oral administration of CHDI-390576 (10, 30, or 100 mg/kg, b.i.d. for 15 days) was well-tolerated with no significant effects on body weight, body temperature, or observational parameters. [1]
Behavioral effects were noted in the open field test. Compared to vehicle control, CHDI-390576 at 30 and 100 mg/kg caused a significant increase in total distance traveled, center distance, and rearing events in both wild-type and R6/2 mice. Average velocity was unchanged. These effects are tentatively attributed to Class IIa HDAC inhibition. [1] |
| Enzyme Assay |
For biochemical HDAC inhibition assays, the catalytic domains of recombinant human HDACs were used. Enzyme activity was measured using fluorogenic peptide substrates: Boc-Lys(TFA)-AMC for Class IIa HDACs (HDAC4, 5, 7, 9) and HDAC8; Ac-Arg-Gly-Lys(Ac)-AMC for HDAC1/2; and Boc-Lys(Ac)-AMC for HDAC3 and HDAC6. Compounds were serially diluted in DMSO and incubated with enzyme and substrate. After a defined period, the reaction was stopped with a developer solution containing trypsin and a deacetylation detection reagent. Fluorescence was measured, and IC₅₀ values were determined from dose-response curves. [1]
Surface plasmon resonance (SPR) was used to determine binding kinetics. The catalytic domain of human HDAC4 was immobilized on a sensor chip. Solutions of the compound at various concentrations were flowed over the chip to measure association and dissociation rates, from which the equilibrium dissociation constant (Kd) and half-life were calculated. [1] |
| Cell Assay |
Cellular Class IIa HDAC inhibition was assessed in Jurkat T cells. Cells were treated with serially diluted compounds for a specified duration. Cellular HDAC activity was measured by adding a cell-permeable, fluorogenic substrate, Boc-Lys(TFA)-AMC. After incubation, fluorescence was quantified to determine the IC₅₀ for intracellular Class IIa HDAC inhibition. [1]
A counter-screen for Class I/IIb HDAC activity was performed in parallel using the substrate Boc-Lys(Ac)-AMC, which is selective for these classes over Class IIa HDACs in the cellular context. [1] Activity against endogenous HDAC substrates was evaluated in A549 cells. Cells were treated with compound, lysed, and protein extracts were analyzed by western blot using antibodies specific for acetylated histone H4 (Lys12) to assess Class I HDAC inhibition, and for acetylated α-tubulin to assess HDAC6 (Class IIb) inhibition. Band intensity was quantified to determine IC₅₀ values. [1] |
| Animal Protocol |
For pharmacokinetic (PK) studies, fed male C57BL/6 mice were administered CHDI-390576 via intravenous injection (5 mg/kg) or oral gavage (10, 30, or 100 mg/kg). For oral administration, the compound was formulated in a suitable vehicle (specific vehicle not detailed in the main text). Blood (for plasma), brain, and muscle tissues were collected at various time points post-dose for bioanalysis. [1]
For the tolerability and behavioral study in R6/2 mice, transgenic mice and wild-type littermates were dosed orally twice daily (b.i.d.) with CHDI-390576 at 10, 30, or 100 mg/kg, or with vehicle, for 15 days. Body weight and temperature were monitored. Open field activity was assessed to measure locomotor and exploratory behavior. [1] |
| ADME/Pharmacokinetics |
After a single oral dose of 10 mg/kg CHDI-390576 in mice, the total plasma exposure (AUC) was 1960 nM·h, the total brain tissue exposure (AUC) was 293 nM·h, and the oral bioavailability was 44% [1]. The volume of distribution (Vss) was 7.3 L/kg, indicating its extensive tissue distribution. The plasma clearance (CL) was 3.1 L/h/kg [1]. The plasma free drug concentration (fu, plasma) was 4.1%, and the brain homogenate free drug concentration (fu, brain) was 0.5%. The brain/plasma free drug partition ratio (Kp, uu, brain) at peak plasma concentration (Cmax) was 0.14 [1]. In the oral dose escalation studies (10, 30, 100 mg/kg), the increase in plasma and brain tissue exposure was approximately proportional to the dose. At doses ≥30 mg/kg, total exposure in plasma and brain tissue exceeded the IC₅₀ of cellular class IIa HDACs within approximately 8 hours. [1]
After oral administration of a 100 mg/kg dose, the estimated free brain tissue Cmax was comparable to the IC₃₅ (35 nM) of cellular class IIa HDACs. [1] The compound showed a high intrinsic clearance rate (383 mL/min/kg) in mouse hepatocytes. [1] Its distribution in blood and plasma was approximately equal. [1] |
| Toxicity/Toxicokinetics |
In IonWorks assays, CHDI-390576 did not show significant inhibition of hERG potassium channels (IC₅₀ > 10 µM, maximum inhibition rate 30%). [1] At a concentration of 50 µM, CHDI-390576 also did not show inhibition of major human CYP450 isoenzymes (1A2, 2C8, 2C9, 2C19, 2D6, 3A4). [1] The compound was negative in the Ames assay, indicating that it does not have potential genotoxicity. [1] In a 15-day repeated-dose study in mice, CHDI-390576 was well tolerated at doses up to 100 mg/kg twice daily without adverse effects on body weight, body temperature or general health. [1]
|
| References | |
| Additional Infomation |
CHDI-390576 is a blood-brain barrier-penetrating diphenylmethylhydroxamic acid inhibitor designed to selectively occupy the catalytic sites of class IIa histone deacetylases (HDAC4, 5, 7, 9). [1] The compound was designed to improve its physicochemical properties (reduced topological polarity surface area, reduced hydrogen bond donors) to minimize the efflux of P-glycoproteins (P-gp) and enhance their exposure in the brain, as demonstrated by its low effective efflux ratio (EER of 1.2) in MDCK-MDR1 cells. [1] The compound was used as a tool compound to evaluate the effects of class IIa HDAC catalytic site occupancy in vitro and in vivo, and to explore its potential applications in the treatment of neurodegenerative diseases such as Huntington's disease and amyotrophic lateral sclerosis (ALS), neuromuscular diseases, heart disease, and cancer. [1]
The co-crystal structure of compound 16 with the catalytic domain of HDAC4 shows that the hydroxamic acid chelates the zinc ion at the active site. The key interactions include the π-π stacking interaction between the pyrimidine end-capping group and Phe871 of HDAC4. [1] |
| Molecular Formula |
C19H13F4N3O2
|
|---|---|
| Molecular Weight |
391.319038152695
|
| Exact Mass |
391.094
|
| CAS # |
1629729-98-1
|
| PubChem CID |
81689842
|
| Appearance |
White to off-white solid powder
|
| LogP |
3.3
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
28
|
| Complexity |
518
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
FC1C=CC=CC=1C(C(NO)=O)C1C=CC(C2N=CC(C(F)(F)F)=CN=2)=CC=1
|
| InChi Key |
LMGDHGQJJLEAPQ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H13F4N3O2/c20-15-4-2-1-3-14(15)16(18(27)26-28)11-5-7-12(8-6-11)17-24-9-13(10-25-17)19(21,22)23/h1-10,16,28H,(H,26,27)
|
| Chemical Name |
2-(2-fluorophenyl)-N-hydroxy-2-[4-[5-(trifluoromethyl)pyrimidin-2-yl]phenyl]acetamide
|
| 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 (In Vitro) |
DMSO : ~250 mg/mL (~638.86 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.32 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 20.8 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: ≥ 2.08 mg/mL (5.32 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 20.8 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 (5.32 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5555 mL | 12.7773 mL | 25.5545 mL | |
| 5 mM | 0.5111 mL | 2.5555 mL | 5.1109 mL | |
| 10 mM | 0.2555 mL | 1.2777 mL | 2.5555 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
