A Study to Evaluate the Pharmacokinetics, Pharmacodynamics and Safety of ASP8232 in Subjects With Renal Impairment and in Type 2 Diabetes Mellitus Subjects With Chronic Kidney Disease

This is a two-part study. Part 1 compares the pharmacokinetics (PK), pharmacodynamics (PD) and safety and tolerability of ASP8232 in healthy subjects with subjects with different degrees of renal impairment; Part 2 is a multiple-dose, placebo-controlled study to evaluate the PK, PD and safety and tolerability of multiple doses of ASP8232 in T2DM subjects with CKD.

Part 1:

Subjects reside in the clinic for 9 days, receiving a single oral dose of ASP8232 on Day 1 under fasted conditions followed by a 168-hours blood and urine PK/PD sampling period. Subjects are discharged on Day 8 and return to the clinic on Days 10, 12, 14, 21, 28, and 42 for the collection of blood PK/PD samples. An End of Study Visit (ESV) takes place after the last PK sample is collected on Day 56.

Part 2:

Subjects are admitted to the clinic on Day -2 in order to collect PD urine samples before dosing begins on Day 1. Subjects receive multiple oral doses of ASP8232 or placebo for 28 days. They are discharged on Day 8 and return to the clinic on Days 14 and 21, and Days 27 to 29 for blood PK/PD and urine PD samples. An ESV takes place 14 to 28 days after the last PK/PD sample is collected.

Main Inclusion: Part 1

• Independent Ethics Committee (IEC)-approved written Informed Consent and privacy language as per national regulations must be obtained from the subject or legally authorized representative prior to any study-related procedures (including withdrawal of prohibited medication, if applicable).
• Male subject and his female spouse/partner who is of childbearing potential must be using highly effective contraception consisting of 2 forms of birth control (1 of which must be a barrier method) starting at screening and continue throughout the study period and for 28 days (or 5 half-lives of the study drug whichever is longer) after final study drug administration.
• Male subject must not donate sperm starting at screening and throughout the study period and for at least 90 days after final study drug administration.
• Female subject must be either:

• post-menopausal (defined as at least one year without any menses) prior to screening, or
• premenarchal prior to screening, or
• documented surgically sterile or status post hysterectomy (at least 1 month before screening), or
• if of childbearing potential, must have a negative urine pregnancy test at screening and must be using highly effective contraception. All females of childbearing potential will be required to use highly effective contraception consisting of 2 forms of birth control (1 of which must be a barrier method) starting at screening and throughout the study period and for 28 days (or 5 half-lives of the study drug whichever is longer) after final study drug administration.
• Female subject must not be lactating, and must not be breast feeding at screening or during the study period and for 28 days [or 5 half-lives of the study drug whichever is longer] after final study drug administration.
• Female subject must not donate ova starting at screening and throughout the study period and for 28 days [or 5 half-lives of the study drug whichever is longer] after final study drug administration.
• Subject agrees not to participate in another interventional study while on treatment.

Healthy Subjects:

• Subject must have pre dose eGFR values (based on the MDRD method) at screening and day -1 higher or equal to 80 mL/min/1.73 m2.

Renal Impaired Subjects:

• Subject must have pre dose eGFR values (based on the MDRD method) at screening and day -1 of 15 to < 30 mL/min/1.73 m2, 30 to

< 60 mL/min/1.73 m2 or 60 to < 80 mL/min/1.73 m2 for severe, moderate or mild renal impaired subjects, respectively.

Part 2 Inclusion:

• Independent Ethics Committee (IEC)-approved written Informed Consent and privacy language as per national regulations must be obtained from the subject or legally authorized representative prior to any study-related procedures (including withdrawal of prohibited medication, if applicable).
• Subject has either known or confirmed T2DM with CKD for at least 1 year [screening].
• Subject is ≥ 35 and ≤ 80 years of age.
• Male subject and his female spouse/partner who is of childbearing potential must be using highly effective contraception consisting of 2 forms of birth control (1 of which must be a barrier method) starting at screening and continue throughout the study period and for 28 days (or 5 half-lives of the study drug whichever is longer) after final study drug administration.
• Male subject must not donate sperm starting at screening and throughout the study period and for at least 90 days after final study drug administration.
• Female subject must be either

• post-menopausal (defined as at least one year without any menses) prior to Screening, or
• premenarchal prior to screening, or
• documented surgically sterile or status post hysterectomy (at least 1 month before screening), or
• if of childbearing potential, must have a negative urine pregnancy test at screening and must be using highly effective contraception1. All females of childbearing potential will be required to use highly effective contraception consisting of 2 forms of birth control (1 of which must be a barrier method) starting at screening and throughout the study period and for 28 days (or 5 half-lives of the study drug whichever is longer) after final study drug administration.
• Female subject must not be lactating, and must not be breast feeding at screening or during the study period and for 28 days (or 5 half-lives of the study drug whichever is longer) after final study drug administration.
• Female subject must not donate ova starting at screening and throughout the study period and for 28 days (or 5 half-lives of the study drug whichever is longer] after final study drug administration.
• Subject is on a stable therapy with ACE inhibitors or ARB for at least 3 months [screening].
• Subject is on a stable anti-hyperglycaemia therapy, e.g. with Metformin, SUD, TZD or DPP-4 inhibitor.
• Subject's eGFR is between 15-60 mL/min/1.73 m2 (based on the MDRD method).
• Subject's HbA1c level is lower than 7.5% at clinic admission on day -2.
• Subject has a stable blood pressure for at least 3 to 6 months prior to enrolment.
• Subject's UACR is higher than 30 mg/g at clinic admission on day -2.

Part 1 Exclusion:

All Subjects:

• Female subject who has been pregnant within 6 months before screening or breast feeding within 3 months before screening.
• Subject has a known or suspected hypersensitivity to ASP8232, or any components of the formulation used.
• Subject has any clinically significant history of allergic conditions (including drug allergies, asthma, eczema, or anaphylactic reactions, but excluding untreated, asymptomatic, seasonal allergies at time of dosing).
• Subject has Gilbert's syndrome.
• Subject has/had febrile illness or symptomatic, viral, bacterial (including upper respiratory infection), or fungal (non-cutaneous) infection within 1 week prior to clinic check in.
• Subject has a history of smoking more than 10 cigarettes (or equivalent amount of tobacco) per day within 3 months prior to admission to the Clinical Unit.
• Subject has a history of drinking more than 21 units of alcohol per week (1 unit = 10 g pure alcohol = 250 mL of beer [5%] or 35 mL of spirits [35%] or 100 mL of wine [12%]) (> 14 units of alcohol for female subjects) within 3 months prior to admission to the Clinical Unit.
• Subject uses drugs of abuse within 3 months prior to admission to the Clinical Unit.
• Subject regularly uses any inducer of metabolism (e.g. barbiturates, rifampin ) in the 3 months prior to admission to the Clinical Unit.
• Subject had any significant blood loss, donated one unit (450 mL) of blood or more, or received a transfusion of any blood or blood products within 60 days or donated plasma within 7 days prior to clinic admission on day -1.
• Subject has positive serology test for Hepatitis B Surface Antigen (HBsAg), anti-Hepatitis A virus (anti-HAV [IgM]), anti-Hepatitis C virus (anti-HCV) or anti- Human immunodeficiency virus 1 + 2 (anti-HIV 1+2).
• Subject participated in any interventional clinical study or has been treated with any investigational drugs within 30 days or 5 half-lives whichever is longer, prior to the initiation of screening.
• Subject is an employee of the Astellas Group or Clinical Research Organization (CRO) involved in the study.

Healthy Subjects:

• Subject has any of the liver function tests (Alanine Aminotransferase [ALT], Aspartate Aminotransferase [AST], total bilirubin [TBL]) above the upper limit of normal. In such a case the assessment may be repeated once [day -1].
• Subject has a mean QTc(F) interval of > 430 ms (for males) and > 450 ms (for females) at screening and day -1. If the mean QTc(F) exceeds the limits above, one additional triplicate electrocardiogram (ECG) can be taken. If this triplicate also gives abnormal result the subject should be excluded.
• Subject uses any prescribed or non-prescribed drugs (including vitamins, hormone replacement therapy, natural and herbal remedies, e.g. St. John's Wort) in the 2 weeks prior to study drug administration, except for occasional use of paracetamol (up to 2 g/day).

Renal Impaired Subjects:

• Subject is on or requires hemodialysis or has received a kidney transplantation.
• Subject has a supine mean systolic blood pressure < 90 or > 160 mmHg and a mean diastolic blood pressure < 50 or > 100 mmHg, or pulse rate < 40 or > 90 beats p/m, on screening and day -1. In such a case the assessment may be repeated once (day -1).
• Subject has a mean QTc(F) interval of > 450 ms (for males) and > 470 ms (for females) at screening and day -1. If the mean QTc(F) exceeds the limits above, one additional triplicate ECG can be taken. If this triplicate also gives abnormal result the subject should be excluded.
• Subject has not been on a stable dose of allowed concomitant medications for at least 2 weeks prior to day 1 and/or for whom dose changes are likely to occur during the study.
• Subject requires or is likely to require any new concomitant medications during the course of the study.
• Subject has obstructive uropathy or other causes of renal impairment not related to parenchymal renal disorder and/or disease of the kidney.
• Subject has renal disease secondary to malignancy.
• Subject has a fluctuating or rapidly deteriorating renal function within 4 weeks prior to screening, as indicated by strongly varying or worsening of clinical and/or laboratory signs of renal impairment within the screening period.
• Subject has any of the liver function tests (ALT, AST, TBL) out of range. In such a case the assessment may be repeated once [day -1].

• ALT or AST > 2 x upper limit of normal (ULN)
• TBL > 1.5 x ULN

Part 2 Exclusion

• Female subject who has been pregnant within 6 months prior to screening assessment or breast feeding within 3 months prior to screening.
• Subject has known or suspected hypersensitivity to ASP8232 or sinistrin, or any components of the formulation used.
• Subject has participated in part 1 of the 8232-CL-0002 study.
• Subject has a mean QTc(F) interval of > 450 ms (for males) and > 470 ms (for females) at screening and day -2. If the mean QTc(F) exceeds the limits above, one additional triplicate ECG can be taken. If this triplicate also gives abnormal result the subject should be excluded.
• Subject has a pulse < 40 or > 90 bpm; mean systolic blood pressure >140 mmHg; mean diastolic blood pressure > 90 mmHg (measurements taken in triplicate after subject has been resting in supine position for 5 min; pulse will be measured automatically) at screening and day -2. In such a case the assessment may be repeated once [day -2].
• Subject is on or requires hemodialysis or has received a kidney transplantation.
• Subject has not been on a stable dose of allowed concomitant medications for at least 2 weeks prior to day 1 and/or for whom dose changes are likely to occur during the study.
• Subject who requires or is likely to require any new concomitant medications during the course of the study.
• Subject who has obstructive uropathy or other causes of renal impairment not related to parenchymal renal disorder and/or disease of the kidney.
• Subject who has renal disease secondary to malignancy.
• Subject who has a fluctuating or rapidly deteriorating renal function within 4 weeks prior to the study, as indicated by strongly varying or worsening of clinical and/or laboratory signs of renal impairment within the screening period.
• Subject has type 1 diabetes mellitus.
• Subject with any of the liver function tests (ALT, AST, TBL) out of range as indicated below. In such a case the assessment may be repeated once [day -2].

• ALT or AST > 2 x ULN
• Total bilirubin > 1.5 x ULN
• Subject has had myocardial infarct or stroke within 6 months prior to screening.
• The subject has Gilbert's Syndrome.
• Subject has any clinically significant history of allergic conditions (including drug allergies, asthma, eczema, or anaphylactic reactions, but excluding untreated, asymptomatic, seasonal allergies at time of dosing).
• The subject has/had febrile illness or symptomatic, viral, bacterial (including upper respiratory infection), or fungal (non-cutaneous) infection within one week prior to clinic admission on day -2.
• Subject has a history of drinking more than 21 units of alcohol per week (1 unit = 10 g pure alcohol = 250 mL of beer [5%] or 35 mL of spirits [35%] or 100 mL of wine [12%]) (> 14 units of alcohol for female subjects) within 3 months prior to admission to the Clinical Unit.
• Subject uses of drugs of abuse within 3 months prior to admission to the Clinical Unit.
• Subject regularly uses any inducer of metabolism (e.g. barbiturates, rifampin) in the 3 months prior to admission to the Clinical Unit.
• Subject had any significant blood loss, donated one unit (450 mL) of blood or more, or received a transfusion of any blood or blood products within 60 days or donated plasma within 7 days prior to clinic admission on day -2.
• Subject has positive serology test for HBsAg, anti HAV (IgM), anti-HCV or anti-HIV 1+2.
• Subject participated in any interventional clinical study or has been treated with any investigational drugs within 30 days or 5 half-lives whichever is longer, prior to the initiation of screening.
• Subject is employee of the Astellas Group or CRO involved in the study.