Quantitation of anacetrapib in human and animal adipose by liquid chromatography with mass spectrometric detection

Abstract

Cholesteryl ester transfer protein (CETP) inhibitor is a target for both lowering low-density lipoproteins and raising high-density lipoproteins. Anacetrapib was the lead compound in our cholesteryl ester transfer protein inhibitor program. Preclinical studies were initiated to support the safety of anacetrapib deposition in adipose tissue, followed by a clinical trial to evaluate the effects of anacetrapib in people with vascular disease. An ultra-high performance liquid chromatography/tandem mass spectrometry method was developed to determine tissue anacetrapib concentrations in the adipose of three animal species and humans. The assays were validated in the concentration ranges of 5–5000 ng/ml and 0.1–100 μg/ml. The anacetrapib concentrations in adipose tissue from preclinical and clinical studies were determined.

Article highlights

Experimental

• A bioanalytical method using LC-MS/MS was developed to quantitatively determine Anacetrapib in adipose tissue. Two different calibration ranges were established for both preclinical and clinical assays based on the dose levels.

• Adipose tissue was homogenized in plasma to ensure uniformity and facilitate liquid-liquid extraction of the drug during sample preparation.

Results

• The method was validated in two different calibration ranges and was successfully implemented for the analysis of Anacetrapib in both preclinical and clinical adipose samples.

• Pharmacokinetic studies of Anacetrapib, with analysis performed on over 100 animal adipose samples from three different species and over 500 human adipose samples were supported.

Keywords: :

• adipose assay
• anacetrapib
• animal adipose tissue
• human adipose tissue
• liquid chromatography (HPLC)
• tandem mass spectrometry (MS/MS)

Financial disclosure

The work described herein was funded by Merck & Co., Rahway, NJ, USA. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Competing interests disclosure

All authors are employees of Merck & Co., Rahway, NJ, USA. The authors have no other competing interests or relevant affiliations with any organization or entity with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The clinical study was approved by National Research Ethics Service Committee South Central – Oxford B. Trial registration number: EudraCT 2010-023467, Date: 2011-01-25. Written consent was obtained from the participants. The preclinical studies were reviewed and approved by the Merck IACUC – Institutionalized Animal Care and Use Committee. The study approval numbers were #12067336020169, #13060026020172, #13020026020099, and #13020026020092.

Data availability statement

The authors certify that this manuscript reports original clinical trial data. The data will not be made publicly available.

Additional information

Funding

The work described herein was funded by Merck & Co., Rahway, NJ, USA.