Primary objective:

To assess the potential effect of oral Clarithromycin on the single-dose pharmacokinetics of Givinostat.

Secondary objective:

To assess the safety and tolerability of concomitant administration of Givinostat plus Clarithromycin.

This study was planned as a phase I, open-label, 3-part, fixed-sequence, non-randomized study in healthy male and female subjects. The study (Part 2) aimed at assessing the potential effect of Clarythromycin on the single dose pharmacokynetics of Givinostat.

The total duration of Part 2 was divided as follows:

• Screening: up to 21 days.
• Treatment Period: Days 1 to 11.
• Safety follow-up visit: 12±2 days.

Subjects were confined at site from Day -1 to Day 11. On Days 1 and 8, a single dose of 50 mg Givinostat, as oral suspension, was administered 1 hour after the planned morning time of Clarithromycin administration.

From Day 4 to Day 10, clarithromycin 500 mg film-coated tablets were administered twice a day, in the morning and in the evening.

The following assessments were performed:

• Blood collection for pharmacokinetic analysis on Days 1 to 4 and 8 to 11.
• Vital signs measurements on Days 1 and 4 to 10.
• 12-lead ECG on Days 1, 3, 7 and 8.
• Blood collection for laboratory tests (hematology and biochemistry) on Day 3.

Subjects were discharged in the morning of Day 11 after completing end of study procedures.

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 tobacco or 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 (see Section 8.5.3).

  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 exclusion criteria:

At Screening

1. Previous use of Givinostat.

2. History of anaphylaxis reaction or clinically significant drug hypersensitivity reaction (e.g., angioedema, Stevens-Johnson syndrome, acute generalized exanthematous pustulosis, drug-induced hypersensitivity syndrome, druginduced neutropenia).

3. Known history of hypersensitivity and/or allergic reactions to Givinostat, histone deacetylases (HDAC) inhibitors or to any excipient in the formulation.

4. History of sorbitol intolerance, sorbitol malabsorption or fructose intolerance.

5. Any medical condition (e.g. gastrointestinal, renal or hepatic, including peptic ulcer, inflammatory bowel disease or pancreatitis) or surgical condition (e.g. cholecystectomy, gastrectomy) that may affect drug pharmacokinetics (absorption, distribution, metabolism or excretion) or subject safety.

6. Systolic blood pressure lower than 90 or over 140 mmHg, diastolic blood pressure lower than 60 or over 90 mmHg, or pulse rate lower than 50 or over 100 bpm.

7. QTcF ˃450 msec.

8. Subjects with history of cardiac arrhythmias (documented), family history of sudden cardiac death or history of additional risk factors for torsades-depointes (e.g. heart failure, hypokalemia, long QT syndrome).

9. Having an estimated glomerular filtration (eGFR) < 90 mL/min, based on creatinine clearance calculation by the Cockcroft-Gault formula and normalized to an average surface area of 1.73 m2.

10. Any of the following abnormal laboratory test values:

    1. Platelet count below the lower limit of the normal range (LLN).
    2. Total white blood cells count below the LLN.
    3. Hemoglobin below the LLN.
    4. Triglycerides above the upper limit of normal range (ULN).
    5. Potassium or magnesium below the LLN.

11. Positive urine alcohol, drugs-of-abuse or cotinine screen tests.

12. Positive serum pregnancy test.

13. If woman, she is breast-feeding.

14. History of alcohol abuse within 1 year prior to screening or regular use of alcohol within 6 months prior to the screening visit (i.e. more than 14 units of alcohol per week for males or more than 7 units for females).

15. History of drug abuse within 1 year prior to screening or use of soft drugs (such as marijuana) within 3 months prior to the screening visit or hard drugs [such as cocaine, phencyclidine (PCP), crack, opioid derivatives including heroin, and amphetamine derivatives] within 1 year prior to screening.

16. Participation in any clinical trial within the previous 2 months.

17. Participation in more than 2 clinical trials within the previous 12 months.

18. Blood donation or significant blood loss (≥ 450 mL) due to any reason or had plasmapheresis within the previous 2 months.

19. Veins unsuitable for intravenous puncture on either arm.

20. Difficulty in swallowing capsules, tablets or suspensions.

21. Any reason which, in the opinion of the Investigator, would prevent the subject from participating in the study.

22. Known history of hypersensitivity and/or allergic reactions to clarithromycin, other macrolides or to any excipient in the formulation.

    At Admission to Treatment Period

23. Any clinically relevant abnormalities on clinical laboratory tests.

24. Positive urine alcohol, drugs-of-abuse or cotinine screen tests.

25. Positive urine pregnancy test.

26. Positive or inconclusive SARS-CoV-2 test prior to admission.

27. Use of prescription or non-prescription medicinal products within the previous 28 days or within 5-half-lives of the medicinal product, whichever is longer.

Twenty (20) subjects received at least one dose of an IMP in the study. They constitute the Safety Analysis Population.

Throughout the study and 10-14 days after the End of Study (follow-up visit), i.e. up to day 30-34, all TEAEs, considered related or not related to IMPs, were collected.

End-of-study was defined as the last study procedure on Day 11 or at early discontinuation of subjects administered at least one dose of investigational product.

Early termination procedures were performed as soon as possible after subject withdrawal, within 10-14 days after last participation of the subject in the study, whenever possible

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