Highly sensitive liquid chromatography-tandem mass spectrometry method for quantification of TAK-448 in human plasma

Abstract

TAK-448 is a nonapeptide analogue and a novel metastin receptor agonist. The aim of this study was to develop a bioanalytical method for TAK-448F (the free base of TAK-448) in human plasma with LC/MS/MS that is sensitive and applicable for the clinical PK studies, and to evaluate the reliability and robustness of the developed method through a validation study in accordance with the regulatory guidance/guideline. The bioanalytical method developed in this study can be outlined as follows. The structural analogue, TAK-683, was used as the internal standard (IS). TAK-448F and the IS were extracted from human plasma using solid phase extraction (SPE) with a polymer-based weak cationic exchanger. After evaporating, the residue was reconstituted and injected into a LC-MS/MS system with ESI probe and analyzed by the selected reaction monitoring (SRM) in the positive ion mode. Separation was performed through an UPLC BEH Phenyl column with the mobile phase of water/methanol/formic acid mixture at a flow rate of 0.2 mL/min. The total run time was 10 minutes. The LLOQ was achieved to be 5 pg/mL with 0.5 mL of human plasma sample. All the validation results met the acceptance criteria in accordance with the regulatory guidance/guideline proving its reliability and robustness. As a result of the clinical study, the human PK profiles of TAK-448F were successfully obtained with this method.

Design and Synthesis of an Investigational Nonapeptide KISS1 Receptor (KISS1R) Agonist, Ac-d-Tyr-Hydroxyproline (Hyp)-Asn-Thr-Phe-azaGly-Leu-Arg(Me)-Trp-NH2 (TAK-448), with Highly Potent Testosterone-Suppressive Activity and Excellent Water Solubility

“…The extremely high pharmacological activity and the excellent solubility of 24 are expected to promote development of sustained-release devices for a novel class of drugs for sex hormone-dependent diseases, for example, prostate cancer. These results of in vitro and in vivo studies strongly suggested that a controlled release of KISS1R agonists suppress the HPG axis to achieve suppression of testosterone concentrations and prompted us to select 24 (Ac- d -Tyr-Hyp-Asn-Thr-Phe-azaGly-Leu-Arg­(Me)-Trp-NH 2 , TAK-448) as a clinical candidate of metastin analogue. − …”

“…Mean pharmacokinetic variable estimates of 1 were as follows: C 5min , 2509 ng/mL; V dss , 1225 mL/kg; and CL total , 819 mL/(h•kg) (Table 3). Area under the plasma concentration−time curve (AUC 0−24h ) values after intravenous injections improved slightly with the replacement of D-Trp 47 in 1 with Hyp (24) or Glu (32). The V dss values for 24 and 32 were 0.3-fold lower than that of 1, probably because of the reduced hydrophobicity.…”

“…Furthermore, 24 was completely dissolved in water up to the concentration of 500 mg/mL. Meanwhile, the Trp 47 (6) and Glu 47 (32) analogues exhibited poor solubility, and the solution turned into gel as with 1. The remarkable difference in water solubility between 1 and 24 with only one amino acid substitution was assumed to be due to the difference in total hydrophobicity and global conformation of these peptides.…”