Potential Clinically Relevant Effects of Sialylation on Human Serum AAG-Drug Interactions Assessed by Isothermal Titration Calorimetry: Insight into Pharmacoglycomics?

Kerep, Robert; Šeba, Tino; Borko, Valentina; Weitner, Tin; Keser, Toma; Lauc, Gordan; Gabričević, Mario

doi:10.3390/ijms24108472

Cited by 3 publications

(1 citation statement)

References 58 publications

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“…Full details of the protein characterization by UPLC-MS have been described elsewhere [ 37 ]. The results were assigned based on N-glycan retention times previously obtained by UPLC-MS/MS analysis [ 38 ]. The assigned peaks were integrated using a custom Python script.…”

Section: Methodsmentioning

confidence: 99%

Influence of Desialylation on the Drug Binding Affinity of Human Alpha-1-Acid Glycoprotein Assessed by Microscale Thermophoresis

Šeba,

Kerep,

Weitner

et al. 2024

Pharmaceutics

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Human serum alpha-1-acid glycoprotein (AAG) is an acute-phase plasma protein involved in the binding and transport of many drugs, especially basic and lipophilic substances. The sialic acid groups that terminate the N-glycan chains of AAG have been reported to change in response to numerous health conditions and may have an impact on the binding of drugs to AAG. In this study, we quantified the binding between native and desialylated AAG and seven drugs from different pharmacotherapeutic groups (carvedilol, diltiazem, dipyridamole, imipramine, lidocaine, propranolol, vinblastine) using microscale thermophoresis (MST). This method was chosen due to its robustness and high sensitivity, allowing precise quantification of molecular interactions based on the thermophoretic movement of fluorescent molecules. Detailed glycan analysis of native and desialylated AAG showed over 98% reduction in sialic acid content for the enzymatically desialylated AAG. The MST results indicate that desialylation generally alters the binding affinity between AAG and drugs, leading to either an increase or decrease in Kd values, probably due to conformational changes of AAG caused by the different sialic acid content. This effect is also reflected in an increased denaturation temperature of desialylated AAG. Our findings indicate that desialylation impacts free drug concentrations differently, depending on the binding affinity of the drug with AAG relative to human serum albumin (HSA). For drugs such as dipyridamole, lidocaine, and carvedilol, which have a higher affinity for AAG, desialylation significantly changes free drug concentrations. In contrast, drugs such as propranolol, imipramine, and vinblastine, which have a strong albumin binding, show only minimal changes. It is noteworthy that the free drug concentration of dipyridamole is particularly sensitive to changes in AAG concentration and glycosylation, with a decrease of up to 15% being observed, underscoring the need for dosage adjustments in personalized medicine.

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Section: Methodsmentioning

confidence: 99%

Influence of Desialylation on the Drug Binding Affinity of Human Alpha-1-Acid Glycoprotein Assessed by Microscale Thermophoresis

Šeba,

Kerep,

Weitner

et al. 2024

Pharmaceutics

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Hollow Fiber-in-Syringe Equilibrium Sampling Through Supported-Liquid Membrane for Evaluation of Drug-Plasma Binding

Barri,

Ramzi,

Idkaidek

et al. 2024

Bioanalysis

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Insights into interspecies protein binding variability using clindamycin as an example

Ahmed,

Böhmdorfer,

Jäger

et al. 2024

Journal of Antimicrobial Chemotherapy

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Background In the preclinical development of new drugs, animal models are often employed to predict their efficacy in humans, relying on translational pharmacokinetic/pharmacodynamic (PK/PD) studies. We performed in vitro experiments focusing on the comparison of plasma protein binding (PPB) and bacterial growth dynamics of clindamycin, a commonly used antimicrobial agent, across a range of drug concentrations and plasma environments. Methods Human, bovine and rat plasma were used for determining PPB of clindamycin at various antibiotic concentrations in buffer and media containing 20% to 70% plasma or pure plasma using ultrafiltration (UF) and equilibrium dialysis (ED). Also bacterial growth and time–kill assays were performed in Mueller–Hinton broth (MHB) containing various percentages of plasma. Results Protein binding of clindamycin correlated well between UF and ED. Notably, clindamycin exhibited substantially lower protein binding to rat plasma compared with human and bovine plasma. Staphylococcus aureus growth was significantly reduced in 70% human, bovine, and rat plasma after 4, 8 and 24 h compared with standard MHB. Time–kill data demonstrated that bacterial counts at both 20% and 70% plasmas were less when compared with MHB at drug concentrations lower than MIC after 4 and 8 h of incubation. For rat plasma, the difference was maintained over 24 h of incubation. Furthermore, a complete bacterial killing at 16 mg/L was observed after 24 h in 20% and 70% human and bovine plasma, but not for rat plasma. Conclusions Recognizing interspecies differences in PB might be essential for optimizing the translational relevance of preclinical studies.

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Potential Clinically Relevant Effects of Sialylation on Human Serum AAG-Drug Interactions Assessed by Isothermal Titration Calorimetry: Insight into Pharmacoglycomics?

Cited by 3 publications

References 58 publications

Influence of Desialylation on the Drug Binding Affinity of Human Alpha-1-Acid Glycoprotein Assessed by Microscale Thermophoresis

Influence of Desialylation on the Drug Binding Affinity of Human Alpha-1-Acid Glycoprotein Assessed by Microscale Thermophoresis

Hollow Fiber-in-Syringe Equilibrium Sampling Through Supported-Liquid Membrane for Evaluation of Drug-Plasma Binding

Insights into interspecies protein binding variability using clindamycin as an example

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