Use of predicted versus measured CCS values from different instrument platforms, and isomer separation on the SELECT SERIES Cyclic IMS

Biotransformation activities require the comparison of metabolites across species and studies. In general, chromatographic retention time, accurate mass measurement and mass spectral data are used to align metabolites. Isomeric metabolite comparison may be more challenging particularly when retention times may differ depending on the analytical conditions used. Additionally, the elemental formulae as well as MS/MS spectra can be identical which significantly increases the complexity of the data interpretation and localization of the biotransformation. The use of collision cross section (CCS) values to compare metabolites analyzed using the SELECT SERIES Cyclic IMS and the SYNAPT G2-Si Q-Tof instruments located in different facilities has been shown here and demonstrates the benefit of such analyte-specific physiochemical property to align metabolites across studies.Moreover, computational prediction of CCS values may provide an additional data asset, allowing the comparison of predicted with measured CCS values. This can further provide additional insights to differentiate between isomers. The prediction can also be used to suggest when additional cyclic ion mobility separation (cIMS) would be beneficial in the separation of isomers and increase confidence in any assignment with the use of higher ion mobility resolution. Examples are given here where cIMS has been used to separate oxygenated metabolites of ranitidine and imipramine. This alternative separation mechanism adds to the separating power of UPLC and is of benefit when isomers co-elute

Potential of measured relative shifts in collision cross section values for biotransformation studies.

Ion mobility spectrometry-mass spectrometry (IMS-MS) separates gas phase ions due to differences in drift time from which reproducible and analyte-specific collision cross section (CCS) values can be derived. Internally conducted in vitro and in vivo metabolism (biotransformation) studies indicated repetitive shifts in measured CCS values (CCSmeas) between parent drugs and their metabolites. Hence, the purpose of the present article was (i) to investigate if such relative shifts in CCSmeas were biotransformation-specific and (ii) to highlight their potential benefits for biotransformation studies. First, mean CCSmeas values of 165 compounds were determined (up to n = 3) using a travelling wave IMS-MS device with nitrogen as drift gas (TWCCSN2, meas). Further comparison with their predicted values (TWCCSN2, pred, Waters CCSonDemand) resulted in a mean absolute error of 5.1%. Second, a reduced data set (n = 139) was utilized to create compound pairs (n = 86) covering eight common types of phase I and II biotransformations. Constant, discriminative, and almost non-overlapping relative shifts in mean TWCCSN2, meas were obtained for demethylation (- 6.5 ± 2.1 Å2), oxygenation (hydroxylation + 3.8 ± 1.4 Å2, N-oxidation + 3.4 ± 3.3 Å2), acetylation (+ 13.5 ± 1.9 Å2), sulfation (+ 17.9 ± 4.4 Å2), glucuronidation (N-linked: + 41.7 ± 7.5 Å2, O-linked: + 38.1 ± 8.9 Å2), and glutathione conjugation (+ 49.2 ± 13.2 Å2). Consequently, we propose to consider such relative shifts in TWCCSN2, meas (rather than absolute values) as well for metabolite assignment/confirmation complementing the conventional approach to associate changes in mass-to-charge (m/z) values between a parent drug and its metabolite(s). Moreover, the comparison of relative shifts in TWCCSN2, meas significantly simplifies the mapping of metabolites into metabolic pathways as demonstrated

Absorption, distribution, metabolism, and excretion (ADME) of 14C-sonidegib (LDE225) in healthy volunteers

Purpose: The absorption, distribution, metabolism, and excretion of the hedgehog pathway inhibitor sonidegib (LDE225) were determined in healthy male subjects. Methods: Six subjects received a single oral dose of 800 mg 14C-sonidegib (74 kBq, 2.0 μCi) under fasting conditions. Blood, plasma, urine, and fecal samples were collected predose, postdose in-house (days 1-22), and during 24-h visits (weekly, days 29-43; biweekly, days 57-99). Radioactivity was determined in all samples using accelerator mass spectrometry (AMS). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to determine concentrations of sonidegib and its main circulating metabolite in plasma. Metabolite profiles and structures were determined in pooled plasma, urine, and fecal samples using high-performance LC-AMS and LC-MS/MS, respectively. Results: A single dose of 14C-sonidegib was well tolerated in healthy subjects. Unchanged sonidegib and total radioactivity reached peak concentration in plasma by 2 and 3 h, respectively, and demonstrated similarly long half-lives of 319 and 331 h, respectively. Absorbed sonidegib (estimated 6-7 %) was extensively distributed, and the approximate terminal volume of distribution was 2,500 L. Unchanged sonidegib and a metabolite resulting from amide hydrolysis were the major circulating components (36.4 and 15.4 % of radioactivity area under the curve, respectively). Absorbed sonidegib was eliminated predominantly through oxidative metabolism of the morpholine part and amide hydrolysis. Unabsorbed sonidegib was excreted through the feces. Metabolites in excreta accounted for 4.49 % of the dose (1.20 % in urine, 3.29 % in feces). The recovery of radioactivity in urine and feces was essentially complete (95.3 ± 1.93 % of the dose in five subjects; 56.9 % of the dose in one subject with incomplete feces collection suspected). Conclusions: Sonidegib exhibited low absorption, was extensively distributed, and was slowly metabolized. Elimination of absorbed sonidegib occurred largely by oxidative and hydrolytic metabolism. © 2014 Springer-Verlag

GAS MEASUREMENTS AT LEA Performance and analysis of labelled biomedical compounds

Few data regarding performance and analysis of labelled biomedical compounds for the internal journal of the ETH Zurich university capturing the collaboration between ETH and Novartis

Evaluation of combustion AMS “MICADAS” for ADME studies

Today, combustion AMS (accelerator mass spectrometry) technology, such as "MICADAS", is considered to enable completion of the human ADME study much earlier and, as a consequence, might represent an opportunity to change the current drug development paradigm. To evaluate the applicability of this method, we successfully performed: a) the validation of MICADAS for radioactivity quantification in biomatrices and, b) a rat ADME study of Leniolisib (CDZ173), where conventional methodology was assessed against a microtracer and “MICADAS” approach

ADME studies of [5-3H]-2′-O-methyluridine nucleoside in mice: A building block in siRNA therapeutics

The chemical modification 2′-O-methyl of nucleosides is often used to increase siRNA stability towards nuclease activities. However, the metabolic fate of modified nucleosides remains unclear. Therefore, the aim of this study was to determine the mass balance, pharmacokinetic, and absorption, distribution, metabolism, and excretion (ADME)-properties of tritium-labeled 2′-O-methyluridine, following a single intravenous dose to male CD-1 mice. The single intravenous administration of [5-3H]-2′-O-methyluridine was well tolerated in mice. Radioactivity was rapidly and widely distributed throughout the body and remained detectable in all tissues investigated throughout the observation period of 48 h. After an initial rapid decline, blood concentrations of total radiolabeled components declined at a much slower rate. [3H]-2′-O-Methyluridine represented a minor component of the radioactivity in plasma (5.89% of [3H]-AUC0-48 h). Three [3H]-2′-O-methyluridine metabolites namely uridine (M1), cytidine (M2), and uracil (M3) were the major circulating components representing 32.8%, 8.11%, and 23.6% of radioactivity area under the curve, respectively. The highest concentrations of total radiolabeled components and exposures were observed in kidney, spleen, pineal body, and lymph nodes. The mass balance, which is the sum of external recovery of radioactivity in excreta and remaining radioactivity in carcass and cage wash, was complete. Renal excretion accounted for about 52.7% of the dose with direct renal excretion of the parent in combination with metabolism to the endogenous compounds cytidine, uracil, cytosine, and cytidine

JDQ443, a Structurally Novel, Pyrazole-Based, Covalent Inhibitor of KRASG12C for the Treatment of Solid Tumors.

Rapid emergence of tumor resistance via RAS pathway reactivation has been reported from clinical studies of covalent KRASG12C inhibitors. Thus, inhibitors with broad potential for combination treatment and distinct binding modes to overcome resistance mutations may prove beneficial. JDQ443 is an investigational covalent KRASG12C inhibitor derived from structure-based drug design followed by extensive optimization of two dissimilar prototypes. JDQ443 is a stable atropisomer containing a unique 5-methylpyrazole core and a spiro-azetidine linker designed to position the electrophilic acrylamide for optimal engagement with KRASG12C C12. A substituted indazole at pyrazole position 3 results in novel interactions with the binding pocket that do not involve residue H95. JDQ443 showed PK/PD activity in vivo and dose-dependent antitumor activity in mouse xenograft models. JDQ443 is now in clinical development, with encouraging early phase data reported from an ongoing Phase Ib/II clinical trial (NCT04699188)