Chemical Analysis of Solid-State Irradiated Human Insulin

Terryn, Herman; Vanhelleputte, Jean-Paul; Maquille, Aubert; Tilquin, Bernard

doi:10.1007/s11095-006-9053-y

Cited by 9 publications

(6 citation statements)

References 25 publications

(17 reference statements)

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“…The CD profile of insulin in the far UV range is shown in Figure 9. As it is evident from this figure, two minima around 208 and 222 nm, which is typical of predominant α‐helix structure of proteins such as insulin110 that was also reported by other researchers,2, 111 was unaffected by the formulation/process and superimposed almost entirely to that of standard insulin profile. This suggested that the protein maintained its α‐helical structure and hence it's secondary structure and physical stability.…”

Section: Resultssupporting

confidence: 81%

A Novel Approach to Prepare Insulin-Loaded Poly (Lactic-Co-Glycolic Acid) Microcapsules and the Protein Stability Study

Emami

Hamishehkar

Najafabadi

et al. 2009

Journal of Pharmaceutical Sciences

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Section: Resultssupporting

confidence: 81%

A Novel Approach to Prepare Insulin-Loaded Poly (Lactic-Co-Glycolic Acid) Microcapsules and the Protein Stability Study

Emami

Hamishehkar

Najafabadi

et al. 2009

Journal of Pharmaceutical Sciences

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“…Dityrosine formation has been observed previously in insulin upon oxidation with H 2 O 2 /peroxidase [15], metal-catalyzed oxidation with H 2 O 2 /Cu [52], ozone [53], carbon electrodes [54], and exposure to γ-radiation [17]. Inter-molecular dityrosine cross-linking is indicated by two of these studies [15], [52], while carbon electrode oxidation suggests intra-molecular dityrosine cross-linking of insulin [54].…”

Section: Discussionsupporting

confidence: 60%

“…Dityrosine is formed upon radical isomerization followed by diradical reaction, and finally enolization [12], [13]. Dityrosine is found in numerous proteins as a result of aging [13], exposure to oxygen free radicals [13], [14], nitrogen dioxide, peroxynitrite, and lipid hydroperoxides [13], enzymatic reaction with peroxidases [13], [15], [16], γ-irradiation [17], and UV-irradiation [18]–[20]. In these cases dityrosine cross-linking (C ortho -C ortho ) can be either intramolecular or intermolecular [2], [12], [20] (see Figure 1B).…”

Section: Introductionmentioning

confidence: 99%

UV-Light Exposure of Insulin: Pharmaceutical Implications upon Covalent Insulin Dityrosine Dimerization and Disulphide Bond Photolysis

et al. 2012

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In this work we report the effects of continuous UV-light (276 nm, ∼2.20 W.m−2) excitation of human insulin on its absorption and fluorescence properties, structure and functionality. Continuous UV-excitation of the peptide hormone in solution leads to the progressive formation of tyrosine photo-product dityrosine, formed upon tyrosine radical cross-linkage. Absorbance, fluorescence emission and excitation data confirm dityrosine formation, leading to covalent insulin dimerization. Furthermore, UV-excitation of insulin induces disulphide bridge breakage. Near- and far-UV-CD spectroscopy shows that UV-excitation of insulin induces secondary and tertiary structure losses. In native insulin, the A and B chains are held together by two disulphide bridges. Disruption of either of these bonds is likely to affect insulin’s structure. The UV-light induced structural changes impair its antibody binding capability and in vitro hormonal function. After 1.5 and 3.5 h of 276 nm excitation there is a 33.7% and 62.1% decrease in concentration of insulin recognized by guinea pig anti-insulin antibodies, respectively. Glucose uptake by human skeletal muscle cells decreases 61.7% when the cells are incubated with pre UV-illuminated insulin during 1.5 h. The observations presented in this work highlight the importance of protecting insulin and other drugs from UV-light exposure, which is of outmost relevance to the pharmaceutical industry. Several drug formulations containing insulin in hexameric, dimeric and monomeric forms can be exposed to natural and artificial UV-light during their production, packaging, storage or administration phases. We can estimate that direct long-term exposure of insulin to sunlight and common light sources for indoors lighting and UV-sterilization in industries can be sufficient to induce irreversible changes to human insulin structure. Routine fluorescence and absorption measurements in laboratory experiments may also induce changes in protein structure. Structural damage includes insulin dimerization via dityrosine cross-linking or disulphide bond disruption, which affects the hormone’s structure and bioactivity.

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“…In a recent paper on the sterilisation of human insulin, protection of the protein was attempted largely in aqueous solution without resort to either lyophilisation or reduction in temperature (Terryn et al 2007) and can be compared with a previous study by the same author in which solid-state insulin was irradiated at radiosterilisation doses (Terryn et al 2006). At 10 kGy, the recovery of insulin was 96.8%.…”

Section: Sterilisation Of Sensitive Biomolecules In Aqueous Solutionmentioning

confidence: 99%

Free radical studies of components of the extracellular matrix: contributions to protection of biomolecules and biomaterials from sterilising doses of ionising radiation

Parsons

2017

Cell Tissue Bank

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The purpose of the current review is show how the principles and techniques of radiation chemistry have enabled the direct reactions of free radicals with biomolecules and biomaterials to be investigated at the molecular level. In particular, the review focusses on the free radical-induced fragmentation of glycosaminoglycans. Glycosaminoglycans are large linear polysaccharides consisting of repeating disaccharide units and are important components of the extracellular matrix (ECM) either in free form (hyaluronan) or as a component of proteoglycans. Oxidative damage of the extracellular matrix components by either enzymatic or non-enzymatic pathways may have implications for the initiation and progression of a range of human diseases. These include arthritis, kidney disease, cardiovascular disease, lung disease, periodontal disease and chronic inflammation. Oxidative damage to hyaluronan by reactive oxidative species and thus the potential mechanism of damage to the ECM and its role in human pathologies is reviewed with particular focus on damage initiated by potential in vivo free radicals such as superoxide, carbonate and hydroxyl radicals. Such knowledge has also allowed radiation protecting systems to be developed so that sterilising doses of radiation can be delivered to sensitive biomolecules such as proteins and glycosaminoglycans, and also to sensitive biomaterials such as tissue allografts.

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Chemical Analysis of Solid-State Irradiated Human Insulin

Abstract: Human insulin samples irradiated in the solid-state at 10 kGy (gamma rays) and 14 kGy (electron-beam) meet the European Pharmacopoeia requirements and can be considered as quite stable towards radiation from a chemical analysis viewpoint.

Cited by 9 publications

References 25 publications

A Novel Approach to Prepare Insulin-Loaded Poly (Lactic-Co-Glycolic Acid) Microcapsules and the Protein Stability Study

A Novel Approach to Prepare Insulin-Loaded Poly (Lactic-Co-Glycolic Acid) Microcapsules and the Protein Stability Study

UV-Light Exposure of Insulin: Pharmaceutical Implications upon Covalent Insulin Dityrosine Dimerization and Disulphide Bond Photolysis

Free radical studies of components of the extracellular matrix: contributions to protection of biomolecules and biomaterials from sterilising doses of ionising radiation

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