Givinostat and Metabolites Pharmacokinetics in Urine and Plasma (Part 3)

An Open-label, Single-center, Study in Healthy Subjects to Evaluate the Plasma and Urine Pharmacokinetics of Givinostat and Its Metabolites Following Single and Multiple Oral Doses of Givinostat (Part 3).

Status

Completed

Phases

Phase 1

Study type

Interventional

Source

ClinicalTrials.gov

Registry ID

NCT05860114

Enrollment

Registered

2023-05-16

Start date

2022-03-21

Completion date

2022-05-24

Last updated

2025-01-24

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

Drug Drug Interaction

Keywords

PK, Pharmacokinetics, Givinostat, Givinostat metabolites

Brief summary

Primary objective: To evaluate the plasma and urine PK of Givinostat following multiple oral doses of Givinostat. Secondary objective: To assess the safety and tolerability multiple oral doses of Givinostat.

Detailed description

This study was planned as a phase I, open-label, 3-part, fixed-sequence, nonrandomized study in healthy male and female subjects. The study (Part 3) aimed at assessing Givinostat and its metabolites (ITF2374, ITF2375, ITF2440 and ITF2563) plasma and urine concentrations and thereof derived pharmacokinetic parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Day 5 to Day 12, with measures reported at Day 13, before single-dose administration). More precisely, Gmean and corresponding 95% CI of Givinostat plasma and urine concentration and Givinostat metabolites plasma concentration versus time profiles following single (Day 1) and multiple-dose (Day 5 to Day 12, with measures reported at Day 13) administration of Givinostat are reported using Linear Scale and Semi-Logarithmic Scale. Descriptive statistics of Givinostat plasma pharmacokinetic parameters and Givinostat metabolites PK parameters following single-dose administration of Givinostat (Day 1) and following multiple-dose administration of Givinostat (Days 5-12, with measures reported at Day 13) were calculated. Descriptive statistics of Givinostat and its metabolites urinary excretion profile following single-dose administration of Givinostat (Day 1) and following multiple dose administration of Givinostat (Days 5-12, with measures reported at Day 13) were calculated. Descriptive statistics of Givinostat and its metabolites cumulative amount of urinary excretion profile following single-dose administration of Givinostat (Day 1) and following multiple-dose administration of Givinostat (Days 5-12, with measures reported at Day 13) were also calculated using linear scales.

Interventions

DRUGGivinostat

Drug: ITF2357 Givinostat 10mg/mL Dose: 10mg/mL; Dosage form: oral suspension On days 1 and 13 givinostat was administered in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. Moreover the subjects were in a semi-recumbent position and remained semi-recumbent until at least 4 hours post-dose.No fluids were allowed from 1 hour before dosing until 2 hours post-dose. Water was provided ad libitum at all other times. From Day 5 to Day 12, subjects received givinostat 50 mg as oral suspension, twice a day, in the morning and in the evening.

Study design

Allocation

Intervention model

SINGLE_GROUP

Primary purpose

TREATMENT

Masking

NONE

Masking description

The study was conducted as open label. Blinding procedures were not applicable.

Eligibility

Sex/Gender

ALL

Age

18 Years to 55 Years

Healthy volunteers

Yes

Inclusion criteria

* A subject was considered eligible for the study if he/she fulfilled all the inclusion criteria: 1. Subject's written informed consent obtained prior to any study-related procedure. 2. Male or female subject, ≥18 and ≤55 years of age, at the time of signing the informed consent. 3. Body mass index (BMI) of 18.5 to 32.0 kg/m2 inclusive, and body weight ≥55 kg and ≤100 kg for females and body weight ≥60 kg and ≤110 kg for males. 4. Non-smoker or ex-smoker (i.e. someone who abstained from using tobaccoor nicotine-containing products for at least 3 months prior to Screening). 5. No clinically relevant diseases. 6. No major surgery within 4 weeks prior to dosing. 7. No clinically relevant abnormalities on physical examination. 8. No clinically relevant abnormalities on 12-lead ECG. 9. No clinically relevant abnormalities on clinical laboratory tests. 10. Negative test results for anti-Human Immunodeficiency virus 1 and 2 antibodies (anti-HIV-1Ab and anti-HIV-2Ab), Hepatitis B surface antigen (HBsAg) and anti-Hepatitis C virus antibodies (anti-HCVAb). 11. Female subjects are eligible if they are of non-childbearing potential or agree to use a non-hormonal highly effective contraceptive method from 28 days prior to Screening until at least 90 days after the last study drug administration. Nonchildbearing potential female is defined as: 1. Menopausal, i.e. no menses for ≥ 12 months without an alternative medical cause other than menopause, and a high FSH level. 2. Pre-menopausal female with documented hysterectomy, bilateral salpingectomy and/or bilateral oophorectomy. A non-hormonal effective contraceptive method is defined as: 1. Intrauterine device. 2. Bilateral tubal occlusion. 3. Total abstinence of heterosexual intercourse, in accordance with the lifestyle of the subject. 4. Vasectomized partner, who has received medical assessment of the surgical success, or clinically diagnosed infertile partner. 12. Male subjects who are sexually active with a female partner of childbearing potential (pregnant or non-pregnant) must use contraception (condom) from investigational product administration up to at least 90 days following the last study drug administration. 13. Male subjects must ensure that his non-pregnant female partner of childbearing potential agrees to consistently and correctly use for the same period a highly effective method of contraception. 14. Male subjects must be willing not to donate sperm until 90 days following the last study drug administration. 15. Willingness and capability to comply with the requirements of the study and ability to understand the study procedures and the risks involved.

Exclusion criteria

A subject was excluded from the study if he/she fulfilled any of the

Design outcomes

Primary

Measure	Time frame	Description
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: CLr/F	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.	Cmax is the Maximum Observed Plasma Concentration. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations using a validated LC-MS/MS analytical method. Summary Statistics for the main PK parameters, like Cmax, Following Single-Dose Administration of Givinostat (Day 1) are reported.
Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.	Tmax is the Time to Maximum Observed Concentration.The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at predose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like tmax, Following Single-Dose Administration of Givinostat (Day 1) are reported.
Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.	Ctrough is the Pre-dose Plasma Concentration. On Day 1 and Day 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.	AUC0-inf is the AUC from Time Zero to Infinity. On Day 1 and Day 13, givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.	Tmax,ss is the Time of occurrence of Cmax,ss. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported in this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for Tmax,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.
Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.	AUC0-t is the AUC from Time Zero to Last Sampling Time with Quantifiable. On Days 1 and 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 h and subjects remained fasted until at least 4 h post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. 48 blood samples were collected in total: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose), * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.	AUC0-τ is the area under the plasma concentration versus time curve. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like AUC0-τ, Following Single-Dose Administration of Givinostat (Day 1) are reported.
Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.	Area under the curve (AUC) is used to describe the total exposure to a drug. The total AUC (AUC0-∞) is the area under the curve from time 0 extrapolated to infinite time. %AUCextrap is the Residual Area or Percentage of Extrapolated Part of AUC0-∞. On Days 1 and 13, givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 h and subjects remained fasted until at least 4 h post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 samples were collected as already described. PK plasma parameters following single oral dose administration of givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.	λz is the Apparent Terminal Elimination Rate Constant. On Day 1 and Day 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.	t1/2 is the Apparent Terminal Half-Life. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like t1/2, Following Single-Dose Administration of Givinostat (Day 1) are reported.
Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.	Vd/F is the Apparent volume of distribution. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like Vd/F, Following Single-Dose Administration of Givinostat (Day 1) are reported.
Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.	Cmax,ss is the Maximum Observed Plasma Concentration at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for Cmax,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.
Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.	AUC0-τ,ss is the AUC during a Dosing Interval at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for AUC0-τ,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.
Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.	CLss/F is the Apparent total body clearance at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for CLss/F, Following Multiple-Dose of Givinostat (Day 13) are reported.
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: Rmax.	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	Rmax is the Maximum urinary excretion rate. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: Tumax	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	tumax is the Time to Rmax. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: AmtCUM	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	AmtCUM is the Cumulative amount of drug excreted in urine. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: AURC0-t	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	AURC0-t is the Area under the urine excretion curve from time zero to last measurable observed excretion rate. The unit of measure is mg because the value measured is an amount. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.
Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: REC%	0-12, 12-24, 24-36, 36-48, 48-72 and 72-96 hours post-dose on days 1 and 13.	REC% is the Percentage of drug recovered in urine. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Secondary

Measure	Time frame	Description
Incidence and Severity of Treatment Emergent Adverse Events	Throughout the study, until 10-14 days after EoS (Day 17), i.e. till Days 27-31	An AE was considered as any untoward medical occurrence in a patient or clinical investigation subject administered a pharmaceutical product and which does not necessarily imply a causal relationship with this treatment. An AE can, therefore, be any unfavorable and unintended sign (including an abnormal laboratory finding, for example), symptom, or disease temporally associated with the use of a medicinal product, whether or not considered related to the medicinal product. The period of observation for the collection of medical occurrences extended from the time when the subject gave Informed Consent until the follow-up visit.

Countries

Portugal

Participant flow

Recruitment details

A total of 11 subjects were screened for participation. Eight (8) subjects (2 women and 6 men) were admitted to study Part 3.

Participants by arm

Arm	Count
Givinostat (Single-dose or Multiple-dose) On day 1, givinostat 50 mg as oral suspension was administered as a single dose, in the morning. On Days 5-12 givinostat 50 mg as oral suspension was administered twice-daily, in the morning and in the evening, as a multiple dose. On day 13 givinostat 50 mg as oral suspension was administered as a single dose, in the morning. Givinostat: Drug: ITF2357 Givinostat 10mg/mL Dose: 10mg/mL; Dosage form: oral suspension	8
Total	8

Arm

Count

Givinostat (Single-dose or Multiple-dose)

On day 1, givinostat 50 mg as oral suspension was administered as a single dose, in the morning. On Days 5-12 givinostat 50 mg as oral suspension was administered twice-daily, in the morning and in the evening, as a multiple dose. On day 13 givinostat 50 mg as oral suspension was administered as a single dose, in the morning. Givinostat: Drug: ITF2357 Givinostat 10mg/mL Dose: 10mg/mL; Dosage form: oral suspension

Total

Baseline characteristics

Characteristic	Givinostat (Single-dose or Multiple-dose)
Age, Categorical <=18 years	0 Participants
Age, Categorical >=65 years	0 Participants
Age, Categorical Between 18 and 65 years	8 Participants
Age, Continuous	30 years STANDARD_DEVIATION 6.4
Race (NIH/OMB) American Indian or Alaska Native	0 Participants
Race (NIH/OMB) Asian	0 Participants
Race (NIH/OMB) Black or African American	0 Participants
Race (NIH/OMB) More than one race	1 Participants
Race (NIH/OMB) Native Hawaiian or Other Pacific Islander	0 Participants
Race (NIH/OMB) Unknown or Not Reported	0 Participants
Race (NIH/OMB) White	7 Participants
Region of Enrollment Portugal	8 participants
Sex: Female, Male Female	2 Participants
Sex: Female, Male Male	6 Participants

Adverse events

Event type	EG000 affected / at risk
deaths Total, all-cause mortality	0 / 8
other Total, other adverse events	7 / 8
serious Total, serious adverse events	0 / 8

Outcome results

Primary

Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax

Cmax is the Maximum Observed Plasma Concentration. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations using a validated LC-MS/MS analytical method. Summary Statistics for the main PK parameters, like Cmax, Following Single-Dose Administration of Givinostat (Day 1) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the Givinostat single-dose administration (Day 1), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Single-Dose Pharmacokinetic Analysis Population.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	48.06 ng/mL	Geometric Coefficient of Variation 25.8
ITF2374	Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	7.34 ng/mL	Geometric Coefficient of Variation 45.7
ITF2375	Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	132.20 ng/mL	Geometric Coefficient of Variation 40.2
ITF2440	Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	88.77 ng/mL	Geometric Coefficient of Variation 16.4
ITF2563	Plasma Pharmacokinetic (PK) Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Cmax	15.38 ng/mL	Geometric Coefficient of Variation 13.8

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ

AUC0-τ is the area under the plasma concentration versus time curve. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like AUC0-τ, Following Single-Dose Administration of Givinostat (Day 1) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	245.73 h*ng/mL	Geometric Coefficient of Variation 23.2
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	52.48 h*ng/mL	Geometric Coefficient of Variation 42.2
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	806.71 h*ng/mL	Geometric Coefficient of Variation 40.6
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	681.56 h*ng/mL	Geometric Coefficient of Variation 25.8
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: AUC0-τ	99.71 h*ng/mL	Geometric Coefficient of Variation 23.1

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2

t1/2 is the Apparent Terminal Half-Life. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like t1/2, Following Single-Dose Administration of Givinostat (Day 1) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	7.16 hours	Geometric Coefficient of Variation 15.7
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	7.40 hours	Geometric Coefficient of Variation 32.8
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	11.21 hours	Geometric Coefficient of Variation 34.4
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	11.65 hours	Geometric Coefficient of Variation 15.4
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: t1/2	12.57 hours	Geometric Coefficient of Variation 27.8

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax

Tmax is the Time to Maximum Observed Concentration.The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at predose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like tmax, Following Single-Dose Administration of Givinostat (Day 1) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.

Arm	Measure	Value (MEDIAN)
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	1.50 hours
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	5.50 hours
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	3.50 hours
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	9.00 hours
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Tmax	10.00 hours

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F

Vd/F is the Apparent volume of distribution. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 22 blood samples were collected as follows: \- 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Days 1, for the determination of Givinostat and its metabolites plasma concentrations. Summary Statistics for the main PK parameters, like Vd/F, Following Single-Dose Administration of Givinostat (Day 1) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	1690.43 Liters	Geometric Coefficient of Variation 21.7
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	0000 Liters	Geometric Coefficient of Variation 0
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	0000 Liters	Geometric Coefficient of Variation 0
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	0000 Liters	Geometric Coefficient of Variation 0
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose Administration of Givinostat: Vd/F	0000 Liters	Geometric Coefficient of Variation 0

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.

AUC0-inf is the AUC from Time Zero to Infinity. On Day 1 and Day 13, givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the givinostat single-dose administration (Day 1) and multiple-dose administration (Day 13), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Single-Dose and Multiple-Dose Pharmacokinetic Analysis Population, respectively.

Arm	Measure	Group	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Single-dose (Day 1)	305.72 h*ng/mL	Geometric Coefficient of Variation 21
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Multiple-dose (Day 13)	562.42 h*ng/mL	Geometric Coefficient of Variation 17.5
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Single-dose (Day 1)	99.01 h*ng/mL	Geometric Coefficient of Variation 49.1
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Multiple-dose (Day 13)	337.71 h*ng/mL	Geometric Coefficient of Variation 53.2
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Single-dose (Day 1)	1239.34 h*ng/mL	Geometric Coefficient of Variation 47.8
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Multiple-dose (Day 13)	2754.89 h*ng/mL	Geometric Coefficient of Variation 77.8
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Multiple-dose (Day 13)	9526.56 h*ng/mL	Geometric Coefficient of Variation 30.2
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Single-dose (Day 1)	2431.31 h*ng/mL	Geometric Coefficient of Variation 12.1
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Single-dose (Day 1)	457.95 h*ng/mL	Geometric Coefficient of Variation 13.4
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-inf.	Multiple-dose (Day 13)	1715.62 h*ng/mL	Geometric Coefficient of Variation 23

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.

AUC0-t is the AUC from Time Zero to Last Sampling Time with Quantifiable. On Days 1 and 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 h and subjects remained fasted until at least 4 h post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. 48 blood samples were collected in total: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose), * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the Givinostat single-dose administration (Day 1 and Day 13) and multiple-dose administration (From Day 5 to Day 12), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Single-Dose and Multiple-Dose Pharmacokinetic Analysis Population, respectively.

Arm	Measure	Group	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Single-dose (Day 1)	300.45 h*ng/mL	Geometric Coefficient of Variation 21.3
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Multiple-dose (Day 13)	554.78 h*ng/mL	Geometric Coefficient of Variation 17.5
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Single-dose (Day 1)	93.30 h*ng/mL	Geometric Coefficient of Variation 51
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Multiple-dose (Day 13)	328.36 h*ng/mL	Geometric Coefficient of Variation 53.9
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Single-dose (Day 1)	1223.60 h*ng/mL	Geometric Coefficient of Variation 47.5
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Multiple-dose (Day 13)	2609.03 h*ng/mL	Geometric Coefficient of Variation 68.2
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Multiple-dose (Day 13)	9262.28 h*ng/mL	Geometric Coefficient of Variation 27.4
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Single-dose (Day 1)	2345.60 h*ng/mL	Geometric Coefficient of Variation 12.7
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Single-dose (Day 1)	439.59 h*ng/mL	Geometric Coefficient of Variation 13
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: AUC0-t.	Multiple-dose (Day 13)	1681.26 h*ng/mL	Geometric Coefficient of Variation 21.8

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap

Area under the curve (AUC) is used to describe the total exposure to a drug. The total AUC (AUC0-∞) is the area under the curve from time 0 extrapolated to infinite time. %AUCextrap is the Residual Area or Percentage of Extrapolated Part of AUC0-∞. On Days 1 and 13, givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 h and subjects remained fasted until at least 4 h post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 samples were collected as already described. PK plasma parameters following single oral dose administration of givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.

Arm	Measure	Group	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Single-dose (Day 1)	1.69 percentage of AUC0-∞	Geometric Coefficient of Variation 21.7
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Multiple-dose (Day 13)	1.33 percentage of AUC0-∞	Geometric Coefficient of Variation 20.6
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Multiple-dose (Day 13)	2.56 percentage of AUC0-∞	Geometric Coefficient of Variation 45.2
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Single-dose (Day 1)	5.39 percentage of AUC0-∞	Geometric Coefficient of Variation 39.8
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Single-dose (Day 1)	0.93 percentage of AUC0-∞	Geometric Coefficient of Variation 97.7
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Multiple-dose (Day 13)	2.13 percentage of AUC0-∞	Geometric Coefficient of Variation 220
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Multiple-dose (Day 13)	1.84 percentage of AUC0-∞	Geometric Coefficient of Variation 112.1
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Single-dose (Day 1)	3.46 percentage of AUC0-∞	Geometric Coefficient of Variation 20.9
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Multiple-dose (Day 13)	1.53 percentage of AUC0-∞	Geometric Coefficient of Variation 82.8
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: %AUCextrap	Single-dose (Day 1)	3.96 percentage of AUC0-∞	Geometric Coefficient of Variation 16.3

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough

Ctrough is the Pre-dose Plasma Concentration. On Day 1 and Day 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the Givinostat single-dose administration (Day 1 and Day 13 in the morning), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Pharmacokinetic Analysis Population.

Arm	Measure	Group	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	single dose (Day 1)	6.75 ng/mL	Geometric Coefficient of Variation 22.3
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	multiple dose (Day 13)	13.16 ng/mL	Geometric Coefficient of Variation 28.7
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	multiple dose (Day 13)	10.18 ng/mL	Geometric Coefficient of Variation 50.8
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	single dose (Day 1)	4.14 ng/mL	Geometric Coefficient of Variation 42.6
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	multiple dose (Day 13)	56.12 ng/mL	Geometric Coefficient of Variation 60.6
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	single dose (Day 1)	28.47 ng/mL	Geometric Coefficient of Variation 53.3
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	multiple dose (Day 13)	279.96 ng/mL	Geometric Coefficient of Variation 15.6
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	single dose (Day 1)	81.58 ng/mL	Geometric Coefficient of Variation 14.9
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	multiple dose (Day 13)	50.11 ng/mL	Geometric Coefficient of Variation 18.8
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-Dose Administration of Givinostat: Ctrough	single dose (Day 1)	14.75 ng/mL	Geometric Coefficient of Variation 11.1

Primary

Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz

λz is the Apparent Terminal Elimination Rate Constant. On Day 1 and Day 13, Givinostat 50 mg as oral suspension was administered as a single dose, in the morning, following an overnight fasting of at least 8 hours and subjects remained fasted until at least 4 hours post-dose. From Day 5 to Day 12 Givinostat 50 mg as oral suspension was administered twice daily. A total of 48 blood samples were collected: * 22 during single-dose treatment (Day 1, at pre-dose and at the timepoints indicated in the timeframe post-dose) and * 26 during the multiple dose treatment (Days 5-12 plus single-dose treatment at Day 13). Of these 26 samples: 22 as per Day 1, and 4 before the morning dose of IMP on Days 9, 10, 11 and 12. PK plasma parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 1 and on Day 13.

Arm	Measure	Group	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Single-dose (Day 1)	0.097 1/hour	Geometric Coefficient of Variation 15.6
Givinostat	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Multiple-dose (Day 13)	0.067 1/hour	Geometric Coefficient of Variation 24.8
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Multiple-dose (Day 13)	0.064 1/hour	Geometric Coefficient of Variation 22.5
ITF2374	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Single-dose (Day 1)	0.094 1/hour	Geometric Coefficient of Variation 33
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Multiple-dose (Day 13)	0.037 1/hour	Geometric Coefficient of Variation 71.6
ITF2375	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Single-dose (Day 1)	0.062 1/hour	Geometric Coefficient of Variation 34.2
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Single-dose (Day 1)	0.060 1/hour	Geometric Coefficient of Variation 15.2
ITF2440	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Multiple-dose (Day 13)	0.047 1/hour	Geometric Coefficient of Variation 49.8
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Multiple-dose (Day 13)	0.052 1/hour	Geometric Coefficient of Variation 31.5
ITF2563	Plasma PK Parameters of Givinostat, Following First Single-dose and Following Multiple-dose Administration of Givinostat: λz	Single-dose (Day 1)	0.055 1/hour	Geometric Coefficient of Variation 27.8

Primary

Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss

AUC0-τ,ss is the AUC during a Dosing Interval at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for AUC0-τ,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the Givinostat multiple-dose administration (Day 13), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Multiple-Dose Pharmacokinetic Analysis Population.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	408.55 h*ng/mL	Geometric Coefficient of Variation 16.2
ITF2374	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	179.63 h*ng/mL	Geometric Coefficient of Variation 41.9
ITF2375	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	1518.21 h*ng/mL	Geometric Coefficient of Variation 50.5
ITF2440	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	3395.41 h*ng/mL	Geometric Coefficient of Variation 15.8
ITF2563	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: AUC0-τ,ss	586.72 h*ng/mL	Geometric Coefficient of Variation 19.3

Primary

Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.

CLss/F is the Apparent total body clearance at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for CLss/F, Following Multiple-Dose of Givinostat (Day 13) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	122.38 L/h	Geometric Coefficient of Variation 16.2
ITF2374	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	0000 L/h	Geometric Coefficient of Variation 0
ITF2375	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	0000 L/h	Geometric Coefficient of Variation 0
ITF2440	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	00000 L/h	Geometric Coefficient of Variation 0
ITF2563	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: CLss/F.	00000 L/h	Geometric Coefficient of Variation 0

Primary

Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss

Cmax,ss is the Maximum Observed Plasma Concentration at steady state. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for Cmax,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.

Population: Eight (8) subjects were enrolled in Part 3 of the study and provided evaluable pharmacokinetic data for the Givinostat single-dose administration (Day 1 and Day 13) and multiple-dose administration (from day 5 to Day 12), without deviations affecting pharmacokinetic interpretation. They constitute the Part 3 Single-Dose and Multiple-Dose Pharmacokinetic Analysis Population, respectively.

Arm	Measure	Value (GEOMETRIC_MEAN)	Dispersion
Givinostat	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	75.50 ng/mL	Geometric Coefficient of Variation 27
ITF2374	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	20.52 ng/mL	Geometric Coefficient of Variation 38.6
ITF2375	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	217.29 ng/mL	Geometric Coefficient of Variation 38.1
ITF2440	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	306.60 ng/mL	Geometric Coefficient of Variation 15.2
ITF2563	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Cmax,ss	55.37 ng/mL	Geometric Coefficient of Variation 20.3

Primary

Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss

Tmax,ss is the Time of occurrence of Cmax,ss. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported in this platform). A total of 26 blood samples were collected as follows: * 22 blood samples at pre-dose and at the timepoints indicated in the timeframe field after the administration of Givinostat (post-dose), on Day 13, for the determination of Givinostat and its metabolites plasma concentrations. * 4 blood samples before the morning dose of on Days 9, 10, 11 and 12, for the determination of pre-dose plasma concentration of Givinostat and its metabolites. Summary Statistics for Tmax,ss, Following Multiple-Dose of Givinostat (Days 5-12 plus single-dose at Day 13, with values calculated at Day 13) are reported.

Time frame: At 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 10, 12, 15, 24, 36, 48, 60, 72, 84 and 96 hours post dose on Day 13.

Arm	Measure	Value (MEDIAN)
Givinostat	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	1.50 hours
ITF2374	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	5.50 hours
ITF2375	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	3.50 hours
ITF2440	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	10.00 hours
ITF2563	Plasma PK Parameters of Givinostat, Following Multiple-dose Administration of Givinostat: Tmax,ss	9.00 hours

Primary

Urinary PK Parameters of Givinostat Following First Single-dose and Following Multiple-dose of Givinostat: AmtCUM

AmtCUM is the Cumulative amount of drug excreted in urine. The Givinostat metabolites were: ITF2374, ITF2375, ITF2440, ITF2563 (not reported on this platform). Urine was collected, beside on Day -1 (baseline), in the following intervals, on Days 1 and 13, for the determination of the amounts of Givinostat and its metabolites excreted in urine: 0h-12h, 12h-24h, 24h-36h, 36h-48h, 48h-72h and 72h-96h post-dose. Givinostat and its metabolites urine levels were measured using a validated LC-MS/MS analytical method. PK urine parameters following single oral dose administration of Givinostat (Day 1) and following multiple oral dose administration of Givinostat (Days 5-12 plus single-dose treatment at Day 13, with values on multiple-dose treatment reported at Day 13) were assessed.