Photodegradation Pathways of Protein Disulfides: Human Growth Hormone

Steinmann, Daniel; Mozziconacci, Olivier; Bommana, Rupesh; Stobaugh, John F.; Wang, Y. John; Schöneich, Christian

doi:10.1007/s11095-017-2256-6

Cited by 18 publications

(28 citation statements)

References 76 publications

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“…Disulfide scrambling induced by UV light exposure has also been observed in a monoclonal antibody and human growth hormone. 18,19 However, it has not previously been reported that photoproducts with a thioether can participate in scrambling due to the initial presence of free thiols from the other sulfur atom in the original disulfide group.…”

Section: Resultsmentioning

confidence: 99%

“…The formation of the dithiohemiacetal group upon photoexcitation of a disulfide bond has previously been identified in a 12 residue model peptide, IgG1 antibody, and human growth hormone. 18,41,42 This diversity in the molecular formats emphasizes that the photoconversion of the disulfide group to a dithiohemiacetal is not a molecule-specific photomodification but relies on fundamental reaction mechanisms induced by UV light. The photoreaction was proposed to be initiated by a direct photoexcitation of the disulfide group which dissociates to thiyl radicals (−S • ) that cause the atomic rearrangements.…”

Section: Resultsmentioning

confidence: 99%

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Characterization of Ultraviolet Photoreactions in Therapeutic Peptides by Femtosecond Laser Catalysis and Mass Spectrometry

et al. 2019

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Peptides and proteins have diverse ultraviolet (UV) photoreaction pathways that can be activated by the energy of the UV photons absorbed. Simple light sources such as lamps are conventionally used to study these photoreactions in solution. This work provides a proof of concept that femtosecond laser technology can function as a highly potent UV source in rapidly conducting UV photostability studies of peptides. Correspondingly, sufficient quantities of photoproducts were generated in 1 min or less, allowing for identification of known and new photomodifications in the therapeutic peptides somatostatin-14 and arginine vasopressin. Identical photoproducts were also generated with a conventional continuous source. The major modifications included N-formylkynurenine, a cross-link between Trp and Phe, a Tyr product with an NH3 loss, and disruption of an unstable disulfide bond into a complex mixture of a trisulfide bond and multiple scrambled dimeric products. In conclusion, femtosecond lasers are extremely useful to drive fast UV-induced reactions for high throughput screening of photostability and modifications in amino acid polymers.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Characterization of Ultraviolet Photoreactions in Therapeutic Peptides by Femtosecond Laser Catalysis and Mass Spectrometry

et al. 2019

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“…A specific improvement for the resolution and detection of these modifications is the use of very long columns (!1m), coupled to mass spectrometry detection. 164,165…”

Section: Biological Oxidation Of Protein Therapeuticsmentioning

confidence: 99%

Proteolysis and Oxidation of Therapeutic Proteins After Intradermal or Subcutaneous Administration

Varkhede

Bommana

Schöneich

et al. 2020

Journal of Pharmaceutical Sciences

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Keywords:intradermal subcutaneous therapeutic proteins PBPK modeling reactive oxygen species proteolysis oxidation lymphatic system a b s t r a c tThe intradermal (ID) and subcutaneous (SC) routes are commonly used for therapeutic proteins (TPs) and vaccines; however, the bioavailability of TPs is typically less than small molecule drugs given via the same routes. Proteolytic enzymes in the dermal, SC, and lymphatic tissues may be responsible for the loss of TPs. In addition, the TPs may be exposed to reactive oxygen species generated in the SC tissue and the lymphatic system in response to injection-related trauma and impurities within the formulation. The reactive oxygen species can oxidize TPs to alter their efficacy and immunogenicity potential. Mechanistic understandings of the dominant proteolysis and oxidative routes are useful in the drug discovery process, formulation development, and to assess the potential for immunogenicity and altered pharmacokinetics (PK). Furthermore, in vitro tools representing the ID or SC and lymphatic system can be used to evaluate the extent of proteolysis of the TPs after the injection and before systemic entry. The in vitro clearance data may be included in physiologically based pharmacokinetic models for improved PK predictions. In this review, we have summarized various physiological factors responsible for proteolysis and oxidation of TPs after ID and SC administration.

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“…While many more spontaneous reactions of amino-acid residues are known than those mentioned, like reversible hydrogen-transfer reactions involving thiyl radicals in Cys-containing peptides and proteins (Mozziconacci et al 2010(Mozziconacci et al , 2011Steinmann et al 2017), still many others are expected to occur, which will need to be detected and explored both mechanistically and biologically (Schöneich 2017). Future research in this field will enormously support the development of safe and potent peptide and protein drugs.…”

Section: Discussionmentioning

confidence: 99%

Susceptibility of protein therapeutics to spontaneous chemical modifications by oxidation, cyclization, and elimination reactions

Grassi

Cabrele

2019

Amino Acids

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Peptides and proteins are preponderantly emerging in the drug market, as shown by the increasing number of biopharmaceutics already approved or under development. Biomolecules like recombinant monoclonal antibodies have high therapeutic efficacy and offer a valuable alternative to small-molecule drugs. However, due to their complex three-dimensional structure and the presence of many functional groups, the occurrence of spontaneous conformational and chemical changes is much higher for peptides and proteins than for small molecules. The characterization of biotherapeutics with modern and sophisticated analytical methods has revealed the presence of contaminants that mainly arise from oxidation-and elimination-prone amino-acid side chains. This review focuses on protein chemical modifications that may take place during storage due to (1) oxidation (methionine, cysteine, histidine, tyrosine, tryptophan, and phenylalanine), (2) intra-and inter-residue cyclization (aspartic and glutamic acid, asparagine, glutamine, N-terminal dipeptidyl motifs), and (3) β-elimination (serine, threonine, cysteine, cystine) reactions. It also includes some examples of the impact of such modifications on protein structure and function.

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Photodegradation Pathways of Protein Disulfides: Human Growth Hormone

Abstract: A comprehensive analysis of photodegradation products revealed a variety of novel photo-products, including cross-links, originating from disulfide degradation. The mechanisms of product formation are discussed.

Cited by 18 publications

References 76 publications

Characterization of Ultraviolet Photoreactions in Therapeutic Peptides by Femtosecond Laser Catalysis and Mass Spectrometry

Characterization of Ultraviolet Photoreactions in Therapeutic Peptides by Femtosecond Laser Catalysis and Mass Spectrometry

Proteolysis and Oxidation of Therapeutic Proteins After Intradermal or Subcutaneous Administration

Susceptibility of protein therapeutics to spontaneous chemical modifications by oxidation, cyclization, and elimination reactions

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