Chemical Instability in Peptide and Protein Pharmaceuticals

Topp, Elizabeth M.; Zhang, Lei; Zhao, Hong; Payne, R. W.; Evans, Gabriel J.; Manning, Mark C.

doi:10.1002/9780470595886.ch2

Cited by 15 publications

(18 citation statements)

References 167 publications

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“…Proteins are prone to alterations in their tertiary fold (physical instabilities, e.g., unfolding and aggregation) and/or modifications in their primary structures (chemical degradation, e.g., oxidation, deamidation, disulfide scrambling, and proteolysis). [19][20][21][22] The risk of such changes is pronounced when the environmental conditions are different from the physiological conditions where the protein has evolved to function. Furthermore, the product protein concentration generally differs markedly from physiological conditions, bringing about additional stress to the protein.…”

Section: Pharmaceutical Protein Formulationsmentioning

confidence: 99%

“…Redox‐active transition metal ions such as Fe, Cu, Mn, and Cr have the potential to initiate chemical degradation, oxidation in particular . These metal ions are often found in pharmaceutical preparations, originated from impurities in excipients (especially in sugars and polymers), or from storage and processing containers (e.g., stainless steel manufacturing vessels) . Phosphate is not generally named as a metal‐chelating excipient for protein formulations, but it might prevent oxidative damage of proteins by binding iron…”

Section: Introductionmentioning

confidence: 99%

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Formulation and Stability of Cytokine Therapeutics

Lipiäinen

Peltoniemi

Sarkhel

et al. 2015

Journal of Pharmaceutical Sciences

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Section: Pharmaceutical Protein Formulationsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Formulation and Stability of Cytokine Therapeutics

Lipiäinen

Peltoniemi

Sarkhel

et al. 2015

Journal of Pharmaceutical Sciences

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“…Conjugation of the fluorophore destabilized the conformational integrity of SM4, and oxidation of the SM4‐AC increased the polarity of the protein and had a more substantial destabilizing influence on the overall stability of the proteins' secondary structure relative to nonoxidized SM4 and SM4‐AC. These observations were not necessarily unexpected, since AC was conjugated to one site on the protein while oxidation is less specific and known to have substantial impacts on the conformation and stability of some proteins, depending on its location within the protein molecule . Furthermore, the heterogeneity of SM4‐AC‐Ox by RP‐HPLC suggests that the measured overall secondary and tertiary structure T m values potentially represented an average of multiple conformations/chemical compositions of the protein within the sample and therefore the stability of each species of oxidized SM4‐AC is potentially different.…”

Section: Discussionmentioning

confidence: 99%

Effect of acrylodan conjugation and forced oxidation on the structural integrity, conformational stability, and binding activity of a glucose binding protein SM4 used in a prototype continuous glucose monitor

et al. 2017

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Continuous glucose monitoring (CGM) devices offer diabetes patients a convenient approach to assist in controlling blood glucose levels. A prototype CGM has been developed that uses the emission profile of a polarity-sensitive fluorophore (acrylodan) conjugated to a glucose/ galactose-binding protein (SM4-AC) to measure the concentration of glucose in vivo. During development, a decrease in the devices signal intensity was observed in vivo over time, which was postulated to be result of oxidative degradation of SM4-AC. A comprehensive physicochemical analysis of SM4-AC was pursued to identify potential mechanisms of signal intensity loss in this CGM during in vitro forced oxidation studies. An assessment of the structural integrity and conformational stability of SM4-AC indicated a relatively decreased polarity and lower tertiary structure stability compared to unconjugated protein (SM4). The stability and polarity of SM4-AC was also altered in the presence of H 2 O 2 . Furthermore, a time-dependent loss in the fluorescence signal of SM4-AC was observed when incubated with H 2 O 2 . An LC-MS peptide mapping analysis of these protein samples indicated that primarily two Met residues in SM4-AC were susceptible to oxidation. When these two residues were genetically altered to an amino acid not prone to Abbreviations: AC, acrylodan; CD, circular dichroism; CGM, Continuous glucose monitor; DPBS, Dulbecco phosphate buffered saline; Fl, Fluorescence; LC-MS, liquid chromatography mass spectrometry; Ox,, Oxidation; RP-HPLC, reversed-phase high performance liquid chromatography; SM4, Glucose/galactose binding protein; T m , thermal melting temperature JMH and NS contributed equally to this work Importance Statement: A comprehensive physicochemical analysis of a sensor (glucose binding protein) used in a prototype continuous glucose monitor was pursued to identify potential mechanisms of signal intensity loss in vivo. The results of this study indicate that genetic alteration of two residues within the protein could increase the long-term stability of this device and therefore improve a patients' diabetes management. oxidation, the glucose binding ability of the protein was retained and no loss of acrylodan fluorescence was observed in the presence of H 2 O 2 . Genetic alteration of these two residues is proposed as an effective approach to increase the long-term stability of SM4-AC within this prototype CGM in vivo.

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“…The presence of water can also facilitate chemical degradation reactions which change the covalent structure of the protein [28]. Hydrolytic processes including deamidation, oxidation and aspartate isomerization may lead to aggregation, denaturation and loss of activity [11,29]. Ideally, the development of stable liquid formulations would be the optimum solution to these problems.…”

Section: Strategies To Improve Protein Stability In the Solid Statementioning

confidence: 97%

Solid-state Protein Formulations

et al. 2015

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When formulated as liquid dosage forms, therapeutic proteins and peptides often show instability during handling as a result of chemical degradation. Solid formulations are frequently required to maintain protein stability during storage, transport and upon administration. Herein we highlight current strategies used to formulate pharmaceutical proteins in the solid form. An overview of the physical instabilities which can arise with proteins is first described. The key solidification techniques of crystallization, freeze-drying and particle forming technologies are then discussed. Examples of current commercial products that are formulated in the solid state are provided and include neutral protamine Hagedorn - insulin crystal suspensions, freeze-dried monoclonal antibodies and leuproride polylactide-co-glycolide microparticles. Finally, future perspectives in solid-state protein formulation are described.

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Chemical Instability in Peptide and Protein Pharmaceuticals

Cited by 15 publications

References 167 publications

Formulation and Stability of Cytokine Therapeutics

Formulation and Stability of Cytokine Therapeutics

Effect of acrylodan conjugation and forced oxidation on the structural integrity, conformational stability, and binding activity of a glucose binding protein SM4 used in a prototype continuous glucose monitor

Solid-state Protein Formulations

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