Skip to Content
Merck
  • A comparison of in vitro ADME properties and pharmacokinetics of azithromycin and selected 15-membered ring macrolides in rodents.

A comparison of in vitro ADME properties and pharmacokinetics of azithromycin and selected 15-membered ring macrolides in rodents.

European journal of drug metabolism and pharmacokinetics (2013-10-12)
Astrid Milić, Vlatka Bencetić Mihaljević, Jovica Ralić, Ana Bokulić, Danijela Nožinić, Branka Tavčar, Boris Mildner, Vesna Munić, Ivica Malnar, Jasna Padovan
ABSTRACT

The purpose of this study was to evaluate the impact of structural modifications on the 15-membered macrolactone ring and/or substituents on the in vitro ADME properties and in vivo pharmacokinetic (PK) profile for selected derivatives in rodents in comparison to azithromycin. Azithromycin and seven selected 15-membered macrolide derivatives, modified either by removal of the sugar moieties, replacement of the amine with a lactam, or addition of lipophilic substituents, were screened in several in vitro ADME assays and in vivo PK studies in rodents. In vitro ADME profiling included assessment of passive permeability and P-gp substrate, metabolic stability in liver microsomes and hepatocytes, as well as CYP direct inhibition measurements. In vivo PK studies were performed in rats (Sprague-Dawley), mice (Balb/c), and P-gp wild-type and deficient mice (CF-1™). Different structural modifications on the azithromycin scaffold resulted in substantial changes in disposition kinetics and oral bioavailability in both rodent species. However, these differences in vivo cannot be predicted based on in vitro results since most of these molecules are classified in the same category. Therefore, in the case of 15-membered ring macrolides, the in vitro ADME screens presented here seem to have low predictive value for in vivo prediction, making their use as routine in vitro screens prior to PK assessments questionable.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Resorufin ethyl ether, ≥98% (TLC), powder
Sigma-Aldrich
Resorufin ethyl ether, suitable for fluorescence, ≥95% (UV)
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
Sigma-Aldrich
Magnesium chloride solution, 0.1 M
Sigma-Aldrich
Methanol, suitable for HPLC, gradient grade, ≥99.9%
Azithromycin, European Pharmacopoeia (EP) Reference Standard
USP
Azithromycin Identity, United States Pharmacopeia (USP) Reference Standard
USP
Azithromycin, United States Pharmacopeia (USP) Reference Standard
Azithromycin for system suitability, European Pharmacopoeia (EP) Reference Standard
Roxithromycin, European Pharmacopoeia (EP) Reference Standard
Azithromycin for peak identification, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Testosterone, European Pharmacopoeia (EP) Reference Standard
Supelco
Testosterone solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Methanol, suitable for HPLC, gradient grade, suitable as ACS-grade LC reagent, ≥99.9%
Sigma-Aldrich
Methanol, suitable for HPLC, ≥99.9%
USP
Ethylene glycol, United States Pharmacopeia (USP) Reference Standard
USP
Sodium bicarbonate, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Magnesium chloride solution, BioUltra, Molecular Biology, ~0.025 M in H2O
Sigma-Aldrich
Testosterone, ≥98%
Supelco
Testosterone, VETRANAL®, analytical standard
Sigma-Aldrich
Ethylene glycol, anhydrous, 99.8%
Sigma-Aldrich
Ethylene glycol, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Ethylene glycol, ReagentPlus®, ≥99%
Sigma-Aldrich
Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol, Absolute - Acetone free
Sigma-Aldrich
Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)