An SPR-based method for Hill coefficient measurements: the case of insulin-degrading enzyme

Distefano, Alessia; Zingale, Gabriele Antonio; Grasso, Giuseppe

doi:10.1007/s00216-022-04122-3

Cited by 4 publications

(4 citation statements)

References 38 publications

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“…Given the lack of binding assay data with varying concentrations of MK-2640 at a fixed [G], h GRI of 1.5 was instead inferred by fitting with the Krug et al’s clinical clamp studies of escalating MK-2640 infusion rates, as seen in Supporting Information, Figure S1. The need for a cooperative h GRI larger than unity is clear, however, even before we attempted to quantitatively fit the model, being (i) evident from the significant initial rise in MK-2640 clearance with increasing concentrations (Supporting Information, Figure S1), and (ii) consistent with literature on insulin-receptor binding . Finally, the mechanistic model outputs a half maximal inhibitory concentration (IC 50 ) of 7.5 mM for glucose, which is in good agreement with the 8 mM reported in experiments.…”

Section: Methodssupporting

confidence: 80%

“…The need for a cooperative h GRI larger than unity is clear, however, even before we attempted to quantitatively fit the model, being (i) evident from the significant initial rise in MK-2640 clearance with increasing concentrations (Supporting Information, Figure S1), and (ii) consistent with literature on insulin-receptor binding. 38 Finally, the mechanistic model outputs a half maximal inhibitory concentration (IC 50 ) of 7.5 mM for glucose, which is in good agreement with the 8 mM reported in experiments.…”

Section: Mechanistic Model Of Competitive Clearance Grissupporting

confidence: 72%

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In Silico Investigation of the Clinical Translatability of Competitive Clearance Glucose-Responsive Insulins

Yang,

Gong

et al. 2023

ACS Pharmacol. Transl. Sci.

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The glucose-responsive insulin (GRI) MK-2640 from Merck was a pioneer in its class to enter the clinical stage, having demonstrated promising responsiveness in in vitro and preclinical studies via a novel competitive clearance mechanism (CCM). The smaller pharmacokinetic response in humans motivates the development of new predictive, computational tools that can improve the design of therapeutics such as GRIs. Herein, we develop and use a new computational model, IM3PACT, based on the intersection of human and animal model glucoregulatory systems, to investigate the clinical translatability of CCM GRIs based on existing preclinical and clinical data of MK-2640 and regular human insulin (RHI). Simulated multi-glycemic clamps not only validated the earlier hypothesis of insufficient glucose-responsive clearance capacity in humans but also uncovered an equally important mismatch between the in vivo competitiveness profile and the physiological glycemic range, which was not observed in animals. Removing the inter-species gap increases the glucose-dependent GRI clearance from 13.0% to beyond 20% for humans and up to 33.3% when both factors were corrected. The intrinsic clearance rate, potency, and distribution volume did not apparently compromise the translation. The analysis also confirms a responsive pharmacokinetics local to the liver. By scanning a large design space for CCM GRIs, we found that the mannose receptor physiology in humans remains limiting even for the most optimally designed candidate. Overall, we show that this computational approach is able to extract quantitative and mechanistic information of value from a posteriori analysis of preclinical and clinical data to assist future therapeutic discovery and development.

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Section: Methodssupporting

confidence: 80%

Section: Mechanistic Model Of Competitive Clearance Grissupporting

confidence: 72%

In Silico Investigation of the Clinical Translatability of Competitive Clearance Glucose-Responsive Insulins

Yang,

Gong

et al. 2023

ACS Pharmacol. Transl. Sci.

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“…We used both wavelengths for our purpose, whereas the two channels could be used independently for parallel experiments, as no subtraction with references is needed in the SPR diffusion experiments (Zingale et al, 2023). The latter were run on a surface which has been previously functionalized with a BSA layer to avoid any interaction of insulin with the gold surface (Distefano et al, 2022). The SPR102‐ AU gold sensor chip is made up of 50 nm of gold and 2 nm of chromium on a glass substrate.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, searching for antifouling surfaces is a very active field of research in the SPR community (Liu et al, 2016). For biomolecules with an isoelectric point (IP) low enough to have a net negative charge on their surfaces at physiological pH, bovine serum albumin (BSA) is often used to minimize nonspecific interactions (Distefano et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Detection of insulin oligomeric forms by a novel surface plasmon resonance‐diffusion coefficient based approach

Calcagno,

Perina,

Zingale

et al. 2024

Protein Science

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Insulin is commonly used to treat diabetes and undergoes aggregation at the site of repeated injections in diabetic patients. Moreover, aggregation is also observed during its industrial production and transport and should be avoided to preserve its bioavailability to correctly adjust glucose levels in diabetic patients. However, monitoring the effect of various parameters (pH, protein concentration, metal ions, etc.) on the insulin aggregation and oligomerization state is very challenging. In this work, we have applied a novel Surface Plasmon Resonance (SPR)‐based experimental approach to insulin solutions at various experimental conditions, monitoring how its diffusion coefficient is affected by pH and the presence of metal ions (copper and zinc) with unprecedented sensitivity, precision, and reproducibility. The reported SPR method, hereby applied to a protein for the first time, besides giving insight into the insulin oligomerization and aggregation phenomena, proved to be very robust for determining the diffusion coefficient of any biomolecule. A theoretical background is given together with the software description, specially designed to fit the experimental data. This new way of applying SPR represents an innovation in the bio‐sensing field and expanding the potentiality of commonly used SPR instruments well over the canonical investigation of biomolecular interactions.

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The Use of Surface Plasmon Resonance to Study the Interactions of Proteins Involved in Conformational Diseases: Experimental Approaches for New Therapeutical Perspectives

Zingale

Distefano

Grasso

2023

CMC

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In recent years, the scientific community has been trying to tackle different diseases by using unifying and holistic approaches based on the concept that it is possible to target apparently very different diseases under a comprehensive general scheme. In other words, various different diseases have been grouped together under the label of “conformational diseases”, because the triggering cause for each malady is the misfolding of a specific protein, whose dyshomeostasis and accumulation cause all the other downhill biomolecular events characteristic of each different disease. In a parallel manner, analytical techniques have developed to investigate protein misfolding and accumulation, so as to give a valid technical support to the investigation of conformational diseases. In this scenario, surface plasmon resonance (SPR) has widely contributed to study many different aspects correlated to conformational diseases, offering the advantages of real time investigations, use of small amounts of biological materials and possibility to mimic the cellular environments without recurring to the use of fluorescent tags. In this review, after a brief introduction about conformational diseases and the SPR technique, a thorough description of the various uses of SPR to investigate the biomolecular mechanisms involved in these diseases is given in order to provide the reader with an exhaustive list as well as a critical perspective of the use of SPR for such topic. The case of Alzheimer’s disease is discussed at a deeper level. We hope that this work will make the reader aware of all the possible SPR experimental approaches, which can be used to develop new possible therapeutic strategies to tackle conformational diseases.

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An SPR-based method for Hill coefficient measurements: the case of insulin-degrading enzyme

Cited by 4 publications

References 38 publications

In Silico Investigation of the Clinical Translatability of Competitive Clearance Glucose-Responsive Insulins

In Silico Investigation of the Clinical Translatability of Competitive Clearance Glucose-Responsive Insulins

Detection of insulin oligomeric forms by a novel surface plasmon resonance‐diffusion coefficient based approach

The Use of Surface Plasmon Resonance to Study the Interactions of Proteins Involved in Conformational Diseases: Experimental Approaches for New Therapeutical Perspectives

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