Biological Activity

Bioactivity / Mechanism

ITF2357 inhibited maize HD2, HD1-B and HD1-A preparations with reported IC50 values of 10, 7.5 and 16 nM, respectively. These cell-free measurements do not establish selectivity among human HDAC isoforms. [1]

In stimulated primary human PBMCs, 100 nM ITF2357 increased acetyl-lysine western-blot signal in a band within the histone-H4 molecular-weight range, with signal retained at 24 h. This provides cellular acetylation evidence without identifying an individual HDAC target. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

Target & Activity

Target	Activity	Value
HD1-B (Cell-free assay)		7.5 nM
HD2 (Cell-free assay)		10 nM
HD1-A (Cell-free assay)		16 nM

In Vitro Studies

Primary human skeletal myoblast cultures exposed to 150 nmol/L givinostat for 24 h showed larger MyHC-positive multinucleated myotubes in representative images than control cultures. The figure provides qualitative morphology evidence; no numerical size effect or replicate count is given. [2]

Inhibition of maize HDAC preparations

Biochemical assay · Cell-free maize HD2, HD1-B and HD1-A preparations [1]

ITF2357 inhibited three maize HDAC preparations; these are not human isoform-selectivity measurements.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Maize HD2 inhibition	IC50 10 nM	[1]
Maize HD1-B inhibition	IC50 7.5 nM	[1]
Maize HD1-A inhibition	IC50 16 nM	[1]

Literature-Reported Treatment Conditions

Method: Maize HDAC inhibition using the method cited to Koelle et al.; Table 1 reports IC50 in nM.
Reported limits: Detailed maize assay implementation and replication are not supplied in Table 1; the cited method paper was not retrieved. The separate PBMC crude-extract protocol is not assigned to this table.

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: ITF2357

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Results: Inhibition of HDAC Activity by ITF2357; Table 1 and footnotes

Maize HD2 inhibition: Table 1 enzyme activity inhibition — Table 1, ITF2357 row, HD2 column; footnote a

Maize HD1-B inhibition: Table 1 enzyme activity inhibition — Table 1, ITF2357 row, HD1-B column; footnote a

Maize HD1-A inhibition: Table 1 enzyme activity inhibition — Table 1, ITF2357 row, HD1-A column; footnote a

Protein acetylation in primary human PBMCs

Cell-based study · Resting human PBMCs from healthy donors, stimulated with LPS [1]

At 100 nM, ITF2357 increased acetyl-lysine immunoreactivity in a protein band in the histone-H4 molecular-weight range. Signal persisted at 24 h; the blot does not identify a particular HDAC isoform.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Acetyl-lysine western-blot signal	Qualitative Increased signal with ITF2357, persisting at 24 h; representative of 1 of 3 donors with similar observations	[1]

Literature-Reported Treatment Conditions

Treatment concentration: 100 nM
Exposure duration: 1 h inhibitor preincubation; sampled after 3, 5, 6 and 24 h of LPS stimulation
Method: Acetyl-lysine western blot of total cellular extracts
Vehicle: Methods specify 0.05% DMSO
Co-treatment: LPS stimulus; SAHA 100 nM comparator
Reported limits: No quantitative acetylation fold change reported for Figure 1.

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: ITF2357

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Figure 1 image/caption; Results: Inhibition of HDAC Activity by ITF2357; Cellular Crude Extract and Acetylation of Proteins methods

Acetyl-lysine western-blot signal: Anti-acetyl-lysine blot; LPS-only lanes and SAHA comparator — Figure 1 image/caption and adjacent Results

Myotube morphology in human skeletal myoblast cultures

Cell-based study · Cultured primary human skeletal myoblasts [2]

Representative MyHC staining showed larger multinucleated myotubes after givinostat exposure than in the control condition; this is qualitative morphology evidence.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
MyHC-positive multinucleated myotube size	Qualitative Larger myotubes than control in representative images	[2]

Literature-Reported Treatment Conditions

Treatment concentration: 150 nmol/L
Exposure duration: 24 h
Method: MyHC immunostaining with DAPI nuclear counterstain
Co-treatment: TSA 50 nmol/L is a separate comparator
Reported limits: Cell supplier, donor number, cell-culture vehicle, replicate count and quantitative size effect are not specified in the read main-text methods/caption.

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: ITF2357

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Results opening paragraph; Figure 1 image/caption

MyHC-positive multinucleated myotube size: MyHC staining; control and separate TSA 50 nmol/L comparator — Figure 1 image/caption; opening Results paragraph

In Vivo Studies

In a 105-day mdx mouse study, daily oral givinostat reduced muscle fibrosis. Functional responses depended on dose and endpoint: 5 mg/kg/day improved treadmill performance at months 1 and 2, whereas a significant reduction in post-exercise diaphragm Evans blue uptake was reported only at 10 mg/kg/day. [2]

In mdx mice receiving an exon-23-targeted palm-tcDNA antisense oligonucleotide, the givinostat co-treatment group had 1.44-fold greater dystrophin protein restoration than the ASO-alone group (p = 0.0076). The 1.21-fold increase in exon skipping was not statistically significant (p = 0.1916). These are combination-treatment comparisons. [3]

In the more severe D2.B10 mouse model, the highest tested givinostat dose in drinking water improved day-105 running distance: 473 ± 22 m versus 296 ± 17 m with vehicle (mean ± s.e.). Histological benefit was time dependent: the authors reported fibrosis reductions at T8 in selected muscles, but no significant fibrosis benefit at T16. [4]

Chronic dose-response in dystrophic mdx mice

Animal study · C57Bl6J mdx mice; treatment began at 1.5 months of age [2]

The 105-day study showed reduced fibrotic area, but functional responses differed by dose and endpoint. Treadmill improvement was most evident at 5 mg/kg/day at months 1 and 2; significant reduction in diaphragm Evans blue uptake was reported only at 10 mg/kg/day.

Conditions shared by the study groups

Route: Oral gavage
Frequency: Daily
Treatment course: 105 d
Vehicle: 0.01% methylcellulose

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
Givinostat 1 mg/kg/day	Methylcellulose alone	Fibrotic area was reduced less than at higher doses. Treadmill performance showed a trend; Evans blue reduction was not statistically significant.	[2]
Givinostat 5 mg/kg/day	Methylcellulose alone	Reduced fibrosis; significantly better treadmill performance at months 1 and 2. Diaphragm Evans blue uptake did not decrease significantly.	[2]
Givinostat 10 mg/kg/day	Methylcellulose alone	Reduced fibrosis and significantly reduced diaphragm Evans blue uptake after exercise; treadmill benefit was less pronounced than at 5 mg/kg/day.	[2]

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: ITF2357

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Animals and In Vivo Treatments; Results associated with Figures 4, 7 and 8; those images/captions; Discussion

Givinostat 1 mg/kg/day · Animals and In Vivo Treatments; Results associated with Figures 4, 7 and 8; those images/captions; Discussion

Givinostat 5 mg/kg/day · Animals and In Vivo Treatments; Results associated with Figures 4, 7 and 8; those images/captions; Discussion

Givinostat 10 mg/kg/day · Animals and In Vivo Treatments; Results associated with Figures 4, 7 and 8; those images/captions; Discussion

Givinostat combined with exon-skipping ASO

Animal study · 8–10-week-old mdx mice (C57BL/10ScSc-Dmdmdx/J); n=7 mice/group [3]

Adding givinostat to palm-tcDNA ASO increased dystrophin protein restoration across muscles, but did not significantly increase exon 23 skipping. The protein response must be attributed to combination treatment.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Dystrophin protein restoration relative to ASO alone across muscles	Fold change 1.44-fold; p = 0.0076 (two-way ANOVA); n=7 mice/group	[3]
Exon 23 skipping relative to ASO alone across muscles	Fold change 1.21-fold; p = 0.1916 (two-way ANOVA); n=7 mice/group	[3]

Conditions shared by the study groups

Route: Givinostat oral gavage; ASO intravenous
Frequency: Givinostat 5 days/week; ASO once/week
Treatment course: 5 weeks HDAC-inhibitor treatment; 4 ASO administrations after first week; tissue harvest 2 weeks after last ASO
Vehicle: Givinostat in water/2% DMSO

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
Givinostat + palm-tcDNA ASO Givinostat 10 mg/kg/day; ASO 25 mg/kg/week	ASO alone; saline-treated mdx and C57BL/10 WT controls also included	Dystrophin protein increased 1.44-fold relative to ASO alone; exon-skipping increase of 1.21-fold was nonsignificant.	[3]

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: Givinostat

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Results: Effect of HDAC inhibitors on Dmd mRNA levels and dystrophin protein levels; Figure 1C–D, Table 1, Figure 2A; Antisense oligonucleotides and animal experiments; Western blot analysis; Statistical analysis

Dystrophin protein restoration relative to ASO alone across muscles: Western blot, normalized to vinculin and tissue-specific WT/mdx calibration; Figure 2 plotted as mean ± SEM — Results: Effect of HDAC inhibitors on dystrophin protein levels; Figure 2A image/table/caption

Exon 23 skipping relative to ASO alone across muscles: TaqMan qRT-PCR; nonsignificant givinostat combination effect — Results: Effect of HDAC inhibitors on Dmd mRNA levels; Table 1; Figure 1D

Givinostat + palm-tcDNA ASO · Results: Effect of HDAC inhibitors on Dmd mRNA levels and dystrophin protein levels; Figure 1C–D, Table 1, Figure 2A; Antisense oligonucleotides and animal experiments; Western blot analysis; Statistical analysis

Chronic treatment in the more severe D2.B10 model

Animal study · Male D2.B10-Dmdmdx/J mice, 6–7 weeks old at purchase; initial groups 12–13 mice; DBA/2J WT comparator [4]

Givinostat at the highest tested drinking-water dose improved day-105 treadmill performance. Histological effects were time dependent: reduced fibrosis was reported in selected muscles at T8, with no significant fibrosis benefit at T16.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Distance run at day 105	Mean ± s.e. 473 ± 22 m (givinostat 37.5 mg/kg/day); 296 ± 17 m (D2.B10 vehicle); 648 ± 31 m (WT)	[4]
Muscle fibrosis at T16	Statistical outcome No significant effect on fibrosis in gastrocnemius, tibialis anterior or diaphragm at T16	[4]

Conditions shared by the study groups

Route: Drinking water
Frequency: Continuous water access; consumption monitored weekly
Treatment course: 105 d
Vehicle: Filtered tap water

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
Givinostat highest tested dose Nominal 37.5 mg/kg/day; estimated ingested range 27.5–41.5 mg/kg/day	Filtered tap water; DBA/2J healthy mice also tested	At day 105, distance run was 473 ± 22 m versus 296 ± 17 m for vehicle (mean ± s.e.). Top dose significantly prolonged time to exhaustion. Fibrosis benefit reported at T8 was absent at T16.	[4]

Givinostat highest tested dose: Reported limits: Nominal dose derived from estimated intake, not a precisely delivered daily amount.

Methods and source locations

Compound reported in the paper: Givinostat (ITF2357); Compound label in the paper: Givinostat

Results apply to the reported experimental material and conditions. No salt conversion, human efficacy inference or SKU lot validation.

Materials and methods: Animals and study design, Drug treatments, Treadmill exercise, Histological analysis, Statistical analysis; Results: Treatment efficacy evaluation in D2.B10 mouse model, Functional activity measurement, Histopathological analysis; Figures 8 and 9

Distance run at day 105: Treadmill exhaustion test; two-way ANOVA with Bonferroni comparisons; text reports significant top-dose benefit; lower doses/steroids nonsignificant — Results: Functional activity measurement, Treadmill exhaustion test; Figure 8A

Muscle fibrosis at T16: Histological fibrosis assessment; 5 mice/group/time point; T8 context in Figure 9 — Results: Histopathological analysis (D2.B10), T16 statements; Histological analysis methods; Figure 9 caption (T8)

Givinostat highest tested dose · Materials and methods: Animals and study design, Drug treatments, Treadmill exercise, Histological analysis, Statistical analysis; Results: Treatment efficacy evaluation in D2.B10 mouse model, Functional activity measurement, Histopathological analysis; Figures 8 and 9

Specifications

Manufacturer	Adooq Bioscience
Actions	Inhibitor
CAS Number	732302-99-7
Applications	HDAC inhibitor
Formula	C₂₄H₂₇N₃O₄.HCl.H₂O
Molecular Weight	476
Synonyms	Gavinostat
SMILES	CC[NH+](CC)CC1=CC2=C(C=C1)C=C(C=C2)COC(=O)NC3=CC=C(C=C3)C(=O)NO.O.[Cl-]

Storage & Solubility

In Vitro Solubility

Reported Solubility

Solvent	Reported Solubility
DMSO	94 mg/mL (197.49 mM)
Water	Insoluble
Ethanol	3 mg/mL

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

In Vivo Preparation

Concentration: 4 mg/mL
Vehicle: 5%DMSO+40%PEG300++5%Tween80+50%ddH2O

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

[1] Leoni F, Fossati G, Lewis EC, et al. The histone deacetylase inhibitor ITF2357 reduces production of pro-inflammatory cytokines in vitro and systemic inflammation in vivo. Molecular medicine (Cambridge, Mass.) (2005). PMID: 16557334 DOI: 10.2119/2006-00005.dinarello Source material
[2] Consalvi S, Mozzetta C, Bettica P, et al. Preclinical studies in the mdx mouse model of duchenne muscular dystrophy with the histone deacetylase inhibitor givinostat. Molecular medicine (Cambridge, Mass.) (2013). PMID: 23552722 DOI: 10.2119/molmed.2013.00011 Source material
[3] Bizot F, Goossens R, Tensorer T, et al. Histone deacetylase inhibitors improve antisense-mediated exon-skipping efficacy in mdx mice. Molecular therapy. Nucleic acids (2022). PMID: 36514350 DOI: 10.1016/j.omtn.2022.11.017 Source material
[4] Licandro SA, Crippa L, Pomarico R, et al. The pan HDAC inhibitor Givinostat improves muscle function and histological parameters in two Duchenne muscular dystrophy murine models expressing different haplotypes of the LTBP4 gene. Skeletal muscle (2021). PMID: 34294164 DOI: 10.1186/s13395-021-00273-6 Source material

Publications

1 publication citing this product

Flavien Bizot, .et al. , Mol Ther Nucleic Acids, 2022, Nov 21;30:606-620 PMID: 36514350

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ITF2357 (Givinostat)

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