Safety and Pharmacokinetics of Single and Multiple Ascending Doses of Avibactam Alone and in Combination with Ceftazidime in Healthy Male Volunteers: Results of Two Randomized, Placebo-Controlled Studies

Henri Merdjan, Manickam Rangaraju, Antoine Tarral

Supplemental Material

Measurement of vital signs

Systolic and diastolic blood pressure and pulse rate were measured using an automated bloodpressure monitoring (dynamap) recorder after 10minutes rest in supine position and after 3 minutes in standing position (except during infusion timewhere the vital signs were measured in supine position only). Where vital signmeasurements were scheduled at the same time as collection of laboratory or pharmacokinetic samples,the blood samplewas withdrawn prior to measurement of vital signs.

Safety and tolerability reporting

Adverse events were classified as mild, moderate or severe according to the following criteria:

  • Mild – awareness of sign or symptom, but easily tolerated.
  • Moderate – discomfort sufficient to cause interference with normal activities.
  • Severe – incapacitating, with inability to perform normal activities.

Analysis Methods

Ceftazidime and avibactam concentrations in the plasma were measured as follows: samples were precipitated with acetonitrile with 1% acetic acid, evaporated to dryness, reconstituted in 1% acetic acid and then injected into the liquid chromatography-tandem mass spectrometry (LC-MS/MS) system, which consisted of a HP1100 binary solvent delivery system (Agilent Technologies, Waldbronn, Germany) with an Atlantis dC18 3 µm analytical column (Waters Corp., Milford, MA, USA) and an API4000 mass spectrometer (Applied Biosystems, Ontario, Canada) with a pneumatic assisted electrospray (TurboIonSprayTM) set to negative mode. Urine samples were first stabilized with 0.01% acetic acid then washed with ethyl acetate. The solvent layer was removed before injecting samples into the LC-MS/MS system. LC for urine samples used an Acquity Binary Solvent Manager (Waters Corp., Milford, MA, USA) with an Atlantis dC18 5 µm column; MS/MS was carried out as for plasma samples. The lower limits of quantitation (LLOQ) in plasma and urine were 0.01 and 0.1 µg/mL, respectively. Analytical performance of the method was assessed using quality control (QC) samples in each analysis batch. Seventy-five percent of the calibration samples were ±15% (±20 % at LLOQ) of their target concentration and at least four of the six QC samples were ±15% of their respective target values. Interbatch precision (coefficient of variation) for avibactam at the low QC level of 25 ng/mL was 6.7% and 6.0%, at the medium QC level of 750 ng/mL it was 5.6% and 7.8%, and at the high QC level of 4,000 ng/mL it was 6.5% and it 6.8%, in plasma and urine QC samples, respectively. These results were considered within the acceptance criteria.

Supplemental Table 1 Observed changes from baseline to End-of-Studyfor serum creatinine and hepatic function laboratory values in the single ascending dose study

Avibactam
dose (mg) / Ceftazidime
dose (mg) / N / Mean change (SD)
Creatinine (µmol/L)
Placebo / 0 / 14 / 3.66 (6.27)
50 / 0 / 8 / 4.68 (5.20)
100 / 0 / 8 / 1.23 (4.27)
250 / 1,000 / 8 / 0.86 (7.94)
500 / 2,000 / 8 / -2.21 (7.90)
1,000 / 0 / 8 / 7.73 (8.42)
1,500 / 0 / 8 / 0.11 (4.63)
2,000 / 0 / 8 / 0.97 (7.47)
Total bilirubin (µmol/L)
Placebo / 0 / 14 / -3.07 (4.40)
50 / 0 / 8 / 0.21 (3.72)
100 / 0 / 8 / -1.47 (4.35)
250 / 1,000 / 8 / -6.23 (8.20)
500 / 2,000 / 8 / -0.68 (6.26)
1,000 / 0 / 8 / 0.86 (5.54)
1,500 / 0 / 8 / -1.28 (3.11)
2,000 / 0 / 8 / -3.44 (3.19)
AST (IU/L)
Placebo / 0 / 14 / -0.9 (6.6)
50 / 0 / 8 / -0.8 (7.2)
100 / 0 / 8 / -0.1 (3.3)
250 / 1,000 / 8 / -0.8 (3.0)
500 / 2,000 / 8 / 1.1 (3.1)
1,000 / 0 / 8 / 1.0 (6.0)
1,500 / 0 / 8 / -3.9 (7.2)
2,000 / 0 / 8 / -2.1 (3.8)
ALT (IU/L)
Placebo / 0 / 14 / 2.6 (4.8)
50 / 0 / 8 / -5.5 (8.2)
100 / 0 / 8 / 0.0 (4.6)
250 / 1,000 / 8 / 0.6 (3.5)
500 / 2,000 / 8 / -1.1 (4.3)
1,000 / 0 / 8 / 0.4 (7.2)
1,500 / 0 / 8 / -2.3 (4.3)
2,000 / 0 / 8 / -2.0 (4.5)
Alkaline phosphatase (IU/L)
Placebo / 0 / 14 / 1.6 (6.2)
50 / 0 / 8 / -0.3 (1.9)
100 / 0 / 8 / 1.8 (6.4)
250 / 1,000 / 8 / 4.0 (6.8)
500 / 2,000 / 8 / -3.8 (8.6)
1,000 / 0 / 8 / 4.4 (5.6)
1,500 / 0 / 8 / 1.8 (11.7)
2,000 / 0 / 8 / 1.6 (6.1)

ALTalanine aminotransferase;ASTaspartate aminotransferase; IUInternational Units; SD, standard deviation

Supplemental Table 2Observed changes from baseline to End-of-Study for serum creatinine and hepatic function laboratory values in the multiple ascending dose part of the second study

Avibactam
dose (mg) / Ceftazidime
dose (mg) / N / Mean (SD)
Creatinine (µmol/L)
Placebo / 0 / 8 / 2.11 (7.45)
500 / 0 / 8 / 1.22 (12.76)
500 / 2000 / 8 / 1.54 (5.97)
750 / 0 / 8 / -1.89 (8.00)
1000 / 0 / 8 / 2.78 (10.36)
Total bilirubin (µmol/L)
Placebo / 0 / 8 / -1.09 (5.55)
500 / 0 / 8 / -2.13 (7.14)
500 / 2000 / 8 / 3.84 (8.40)
750 / 0 / 8 / -1.53 (4.86)
1000 / 0 / 8 / -1.51 (4.55)
AST (IU/L)
Placebo / 0 / 8 / -0.1 (4.1)
500 / 0 / 8 / 3.5 (6.5)
500 / 2000 / 8 / 10.1 (11.2)
750 / 0 / 8 / 7.1 (10.5)
1000 / 0 / 8 / -4.4 (4.9)
ALT (IU/L)
Placebo / 0 / 8 / 7.1 (10.6)
500 / 0 / 8 / 10.4 (12.4)
500 / 2000 / 8 / 26.1 (18.9)
750 / 0 / 8 / 9.0 (11.6)
1000 / 0 / 8 / 2 (10.3)
Alkaline phosphatase (IU/L)
Placebo / 0 / 8 / 2.8 (5.7)
500 / 0 / 8 / -1.6 (5.4)
500 / 2000 / 8 / -1.8 (6.7)
750 / 0 / 8 / -1.5 (1.8)
1000 / 0 / 8 / 0.3 (5.1)

ALTalanine aminotransferase;ASTaspartate aminotransferase; IUInternational Units; SD, standard deviation

Supplemental Table 3Observed serum creatinine and hepatic function laboratory values in the absolute oral bioavailability part of the second study (n = 8)

Laboratory parameter / Mean (SD)
Creatinine (µmol/L)
Screening / 85.41 (11.08)
Period1 Day-1 / 85.06 (12.01)
Period2 Day-1 / 86.31 (10.73)
End-Of-Study / 88.40 (14.78)
Total bilirubin (µmol/L)
Screening / 9.63 (2.76)
Period1 Day-1 / 9.44 (2.05)
Period2 Day-1 / 9.20 (2.61)
End-Of-Study / 8.56 (4.50)
AST (IU/L)
Screening / 25.9 (6.3)
Period1 Day-1 / 22.0 (5.8)
Period2 Day-1 / 22.5 (5.1)
End-Of-Study / 23.0 (4.5)
ALT (IU/L)
Screening / 20.6 (7.6)
Period1 Day-1 / 17.9 (7.4)
Period2 Day-1 / 18.9 (6.4)
End-Of-Study / 19.3 (6.1)

ALTalanine aminotransferase;ASTaspartate aminotransferase; IUInternational Units; SD, standard deviation.

References ranges: creatinine = 44.2–115.1 µmol/L; total bilirubin ≤22.2 µmol/L; AST = 5–50 IU/L;
ALT 5–50IU/L.

Supplemental Figure 1.Study design of the single ascending dose (SAD) study.

AVI, avibactam; CAZ, ceftazidime

Supplemental Figure 2.Study design of the multiple ascending dose (MAD) part of the second study.

AVI, avibactam; CAZ, ceftazidime; IV, intravenous; q8h, every 8 hours

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