Abstract
Peptides are small polymers composed of 40 or fewer amino acids and are an increasingly important class of drugs. Therapeutic peptides are less immunogenic and more economical than biologics while offering greater safety, selectivity, efficacy, and specificity than small molecule drugs. Despite this, they are challenging to mold in a stable formulation due to their susceptibility to degradation. By understanding the degradation behavior of such peptide drugs, researchers and pharmaceutical manufacturers can design safe, effective, and stable peptide formulations. From a scientific standpoint, forced degradation studies are an indispensable tool to forecast the stability of any molecule during its development phase. Being structurally diverse, the degradation of peptide drugs is different from the small molecules. This review provides a practical summary of strategies adopted to perform the stress stability testing for different peptide therapeutics including a selection of stress conditions, degradation products formed, and an analytical methodology used for the characterization of degradation products. Hence, it will help in developing a protocol for performing forced degradation studies on peptide therapeutics of interest. In-depth discussions of the different peptide degradation mechanisms are also included, along with preventive measures. The information presented here in the form of case studies on the degradation profile of existing peptide drugs will help in designing more resistant peptide drugs by chemical modifications, as well as aid in the advancement of generic peptide drug product development. In brief, this review presents the way of controlling peptide degradation from synthesis to formulation development based on their constituted amino acids.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Ala:
-
Alanine
- ANDA:
-
Abbreviated New Drug Application
- APCI:
-
Atmospheric Pressure Chemical Ionization
- Asn:
-
Asparagine
- Asp:
-
Aspartic acid
- D:
-
Dextrorotary
- Cys:
-
Cysteine
- DESI:
-
Desorption Electrospray Ionization
- EMA:
-
European Medicines Agency
- ELDI:
-
Electrospray Laser Desorption Ionization
- ESI:
-
Electro Spray Ionization
- FDA:
-
Food and Drug Administration
- FRET:
-
Fluorescence Resonance Energy Transfer
- FTICR:
-
Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry
- Gln:
-
Glutamine
- GLP-1:
-
Glucagon-like peptide 1
- Glu:
-
Glutamic acid
- Gly:
-
Glycine
- His:
-
Histidine
- HIV:
-
Human Immunodeficiency Virus
- HPLC:
-
High-Performance Liquid Chromatography
- HRMS:
-
High-Resolution Mass Spectrometry
- HAS:
-
Human Serum Albumin
- ICH:
-
International Council for Harmonisation
- IEX:
-
Ion Exchange Chromatography
- Ile:
-
Isoleucine
- L:
-
Levorotary
- Lys:
-
Lysine
- MALDI:
-
Matrix-Assisted Laser Desorption/Ionization
- MS:
-
Mass Spectrometry
- Met:
-
Methionine
- NDA:
-
New Drug Application
- NFK:
-
N-formylkynurenine
- Nle:
-
Norleucine
- NMR:
-
Nuclear Magnetic Resonance
- OGD:
-
Office of Generic Drugs
- PEG:
-
Polyethylene Glycol
- PEO:
-
Polyethylene Oxide
- Phe:
-
Phenylalanine
- Pro:
-
Proline
- QTOF:
-
Quadrupole Time-of-Flight Mass Spectrometry
- rDNA:
-
Recombinant DNA
- RLD:
-
Reference Listed Drug
- SEC:
-
Size-Exclusion Chromatography
- Ser:
-
Serine
- TOF:
-
Time-of-Flight
- Trp:
-
Tryptophan
- Tyr:
-
Tyrosine
- UHPLC:
-
Ultra-Performance Liquid Chromatography
- Xaa:
-
Any amino acid
References
Abrams C (2009) New advances in the treatment of adult chronic immune thrombocytopenic purpura: role of thrombopoietin receptor-stimulating agents. Biol Targets Ther. https://doi.org/10.2147/BTT.2009.3803
Aeschbach R, Amadoò R, Neukom H (1976) Formation of dityrosine cross-links in proteins by oxidation of tyrosine residues. Biochim Biophys Acta - Protein Struct 439:292–301. https://doi.org/10.1016/0005-2795(76)90064-7
Agon VV, Bubb WA, Wright A, Hawkins CL, Davies MJ (2006) Sensitizer-mediated photooxidation of histidine residues: Evidence for the formation of reactive side-chain peroxides. Free Radic Biol Med 40:698–710. https://doi.org/10.1016/j.freeradbiomed.2005.09.039
Akbarian M, Khani A, Eghbalpour S, Uversky VN (2022) Bioactive Peptides: Synthesis, Sources, Applications, and Proposed Mechanisms of Action. Int J Mol Sci 23:1445. https://doi.org/10.3390/ijms23031445
Alsante K, Ando A, Brown R, Ensing J, Hatajik T, Kong W, Tsuda Y (2007) The role of degradant profiling in active pharmaceutical ingredients and drug products☆. Adv Drug Deliv Rev 59:29–37. https://doi.org/10.1016/j.addr.2006.10.006
Angelova A, Drechsler M, Garamus VM, Angelov B (2019) Pep-Lipid Cubosomes and Vesicles Compartmentalized by Micelles from Self-Assembly of Multiple Neuroprotective Building Blocks Including a Large Peptide Hormone PACAP-DHA. ChemNanoMat 5:1381–1389. https://doi.org/10.1002/cnma.201900468
Ashraf CM, Ahmad I, Lugemwa FKN (1980) Kinetics and mechanism of the oxidation of L-phenylalanine by hydrogen peroxide in the presence of ferrous sulfate as a catalyst. J Org Chem 45:3202–3204. https://doi.org/10.1021/jo01304a012
Barany G, Merrifield RB (1977) A new amino protecting group removable by reduction. Chemistry of the dithiasuccinoyl (Dts) function. J Am Chem Soc 99:7363–7365. https://doi.org/10.1021/ja00464a050
Bellmaine S, Schnellbaecher A, Zimmer A (2020) Reactivity and degradation products of tryptophan in solution and proteins. Free Radic Biol Med 160:696–718. https://doi.org/10.1016/j.freeradbiomed.2020.09.002
Benet A, Halseth T, Kang J, Kim A, Ackermann R, Srinivasan S, Schwendeman S, Schwendeman A (2021) The Effects of pH and Excipients on Exenatide Stability in Solution. Pharmaceutics 13:1263. https://doi.org/10.3390/pharmaceutics13081263
Benham CJ, Saleet Jafri M (1993) Disulfide bonding patterns and protein topologies. Protein Sci 2:41–54. https://doi.org/10.1002/pro.5560020105
Bhavsar S, Mudaliar S, Cherrington A (2013) Evolution of Exenatide as a Diabetes Therapeutic. Curr Diabetes Rev 9:161–193. https://doi.org/10.2174/1573399811309020007
Bijarnia RK, Bachtler M, Chandak PG, van Goor H, Pasch A (2015) Sodium thiosulfate ameliorates oxidative stress and preserves renal function in hyperoxaluric rats. PLoS ONE 10:e0124881. https://doi.org/10.1371/journal.pone.0124881
Bonifačić M, Hug GL, Schöneich C (2000) Kinetics of the Reactions between Sulfide Radical Cation Complexes, [S∴S] + and [S∴N] +, and Superoxide or Carbon Dioxide Radical Anions. J Phys Chem A 104:1240–1245. https://doi.org/10.1021/jp9934578
Böttger R, Hoffmann R, Knappe D (2017) Differential stability of therapeutic peptides with different proteolytic cleavage sites in blood, plasma and serum. PLoS ONE 12:e0178943. https://doi.org/10.1371/journal.pone.0178943
Broberg A, Nord C, Levenfors JJ, Bjerketorp J, Guss B, Öberg B (2021) In-peptide amino acid racemization via inter-residue oxazoline intermediates during acidic hydrolysis. Amino Acids 53:323–331. https://doi.org/10.1007/s00726-021-02951-7
Buckley ST, Hubálek F, Rahbek UL (2016) Chemically modified peptides and proteins - critical considerations for oral delivery. Tissue Barriers 4:e1156805. https://doi.org/10.1080/21688370.2016.1156805
Castaño C, Oliveros E, Thomas AH, Lorente C (2015) Journal of photochemistry & photobiology, b : biology histidine oxidation photosensitized by pterin : pH dependent mechanism. JPB 153:483–489. https://doi.org/10.1016/j.jphotobiol.2015.10.026
Castaño C, Vignoni M, Vicendo P, Oliveros E, Thomas AH (2016) Degradation of tyrosine and tryptophan residues of peptides by type I photosensitized oxidation. J Photochem Photobiol B Biol 164:226–235. https://doi.org/10.1016/j.jphotobiol.2016.09.024
Catak S, Monard G, Aviyente V, Ruiz-López MF (2009) Deamidation of asparagine residues: direct hydrolysis versus succinimide-mediated deamidation mechanisms. J Phys Chem A 113:1111–1120. https://doi.org/10.1021/jp808597v
Cavaco M, Andreu D, Castanho MARB (2021) The challenge of peptide proteolytic stability studies: scarce data, difficult readability, and the need for harmonization. Angew Chemie 133:1710–1712. https://doi.org/10.1002/ange.202006372
Cervantes C, Mora JR, Rincón L, Rodríguez V (2020) Theoretical study of the mechanism of 2,5-diketopiperazine formation during pyrolysis of proline. Mol Phys 118:e1594422. https://doi.org/10.1080/00268976.2019.1594422
Chelius D, Jing K, Lueras A, Rehder DS, Dillon TM, Vizel A, Rajan RS, Li T, Treuheit MJ, Bondarenko PV (2006) Formation of Pyroglutamic Acid from N-Terminal Glutamic Acid in Immunoglobulin Gamma Antibodies. Anal Chem 78:2370–2376. https://doi.org/10.1021/ac051827k
Coohill TP, Sagripanti J-L (2008) Overview of the inactivation by 254nm ultraviolet radiation of bacteria with particular relevance to biodefense. Photochem Photobiol. https://doi.org/10.1111/j.1751-1097.2008.00387.x
Costantino HR, Langer R, Klibanov AM (1994) Moisture-induced aggregation of lyophilized insulin. Pharm Res. https://doi.org/10.1023/A:1018981208076
Costantino HR, Culley H, Chen L, Morris D, Houston M, Roth S, Phoenix MJ, Foerder C, Philo JS, Arakawa T, Eidenschink L, Andersen NH, Brandt G, Quay SC (2009) Development of calcitonin salmon nasal spray: similarity of peptide formulated in chlorobutanol compared to benzalkonium chloride as preservative. J Pharm Sci 98:3691–3706. https://doi.org/10.1002/jps.21690
Cromwell MEM, Hilario E, Jacobson F (2006) Protein aggregation and bioprocessing. AAPS J 8:E572–E579. https://doi.org/10.1208/aapsj080366
D’Hondt M, Fedorova M, Peng C-Y, Gevaert B, Taevernier L, Hoffmann R, De Spiegeleer B (2014) Dry heat forced degradation of buserelin peptide: kinetics and degradant profiling. Int J Pharm 467:48–59. https://doi.org/10.1016/j.ijpharm.2014.03.043
Dang T, Süssmuth RD (2017) Bioactive peptide natural products as lead structures for medicinal use. Acc Chem Res 50:1566–1576. https://doi.org/10.1021/acs.accounts.7b00159
Davies MJ (2016) Protein oxidation and peroxidation. Biochem J 473:805–825. https://doi.org/10.1042/BJ20151227
Debnath, D., Cheriyath, P., 2022. Octreotide, StatPearls.
Delmar JA, Buehler E, Chetty AK, Das A, Quesada GM, Wang J, Chen X (2021) Machine learning prediction of methionine and tryptophan photooxidation susceptibility. Mol Ther - Methods Clin Dev 21:466–477. https://doi.org/10.1016/j.omtm.2021.03.023
Dion MZ, Leiske D, Sharma VK, Zuch de Zafra CL, Salisbury CM (2018) Mitigation of oxidation in therapeutic antibody formulations: a biochemical efficacy and safety evaluation of N-acetyl-tryptophan and L-methionine. Pharm Res 35:222. https://doi.org/10.1007/s11095-018-2467-5
Doulou E, Kalomiraki M, Parla A, Thermos K, Chaniotakis NA, Panderi I (2021) Hydrophilic interaction liquid chromatography coupled with fluorescence detection (hilic-fl) for the quantitation of octreotide in injection forms. Analytica 2:121–129. https://doi.org/10.3390/analytica2040012
Dyson HJ, Wright PE (1993) Peptide conformation and protein folding. Curr Opin Struct Biol 3:60–65. https://doi.org/10.1016/0959-440X(93)90203-W
Eronina TB, Chebotareva NA, Kleymenov SY, Roman SG, Makeeva VF, Kurganov BI (2010) Effect of 2-hydroxypropyl-β-cyclodextrin on thermal stability and aggregation of glycogen phosphorylase b from rabbit skeletal muscle. Biopolymers 93:986–993. https://doi.org/10.1002/bip.21508
Evans RM, Krahn N, Murphy BJ, Lee H, Armstrong FA, Söll D (2021) Selective cysteine-to-selenocysteine changes in a [NiFe]-hydrogenase confirm a special position for catalysis and oxygen tolerance. Proc Natl Acad Sci USA 118:e2100921118. https://doi.org/10.1073/pnas.2100921118
Felix A, Moroder L, Toniolo C (2004) 6.7 Pyroglutamic Acid Peptides. In: Weyl H (ed) Methods of Organic Chemistry, 4th edn. Georg Thieme Verlag, Stuttgart
U.S. Food and Drug Administration, C. for drug evaluation, 2021. ANDAs for Certain Highly Purified Synthetic Peptide Drug Products That Refer to Listed Drugs of rDNA Origin Guidance for Industry ANDAs for Certain Highly Purified Synthetic Peptide Drug Products That Refer to Listed Drugs of rDNA Origin Guidance for Indu.
Fransson JR (1997) Oxidation of human insulin-like growth factor i in formulation studies. 3. Factorial experiments of the effects of ferric ions, EDTA, and visible light on methionine oxidation and covalent aggregation in aqueous solution. J Pharm Sci 86:1046–1050. https://doi.org/10.1021/js960484q
Fricker L (2007) Carboxypeptidases. In: Enna SJ, Bylund DB (eds) Elsevier Science (Firm), XPharm: the comprehensive pharmacology reference. Elsevier, New York, pp 1–4
Galande AK, Trent JO, Spatola AF (2003) Understanding base-assisted desulfurization using a variety of disulfide-bridged peptides. Biopolymers 71:534–551. https://doi.org/10.1002/bip.10532
Gammelgaard SK, Petersen SB, Haselmann KF, Nielsen PK (2019) Characterization of ultraviolet photoreactions in therapeutic peptides by femtosecond laser catalysis and mass spectrometry. ACS Omega 4:14517–14525. https://doi.org/10.1021/acsomega.9b01749
Garton M, Nim S, Stone TA, Wang KE, Deber CM, Kim PM (2018) Method to generate highly stable D-amino acid analogs of bioactive helical peptides using a mirror image of the entire PDB. Proc Natl Acad Sci USA 115:1505–1510. https://doi.org/10.1073/pnas.1711837115
Geiger T, Clarke S (1987) Deamidation, isomerization, and racemization at asparaginyl and aspartyl residues in peptides. Succinimide-linked reactions that contribute to protein degradation. J Biol Chem 262:785–794. https://doi.org/10.1016/s0021-9258(19)75855-4
Gentilucci L, De Marco R, Cerisoli L (2010) Chemical modifications designed to improve peptide stability: incorporation of non-natural amino acids, pseudo-peptide bonds, and cyclization. Curr Pharm Des 16:3185–3203. https://doi.org/10.2174/138161210793292555
Girija KS, Kasimala BB, Anna VR (2021) A new high-performance liquid chromatography method for the separation and simultaneous quantification of eptifibatide and its impurities in pharmaceutical injection formulation. Int J Appl Pharm 13:165–172
Gorris HH, Bade S, Röckendorf N, Albers E, Schmidt MA, Fránek M, Frey A (2009) Rapid Profiling of Peptide Stability in Proteolytic Environments. Anal Chem 81:1580–1586. https://doi.org/10.1021/ac802324f
Gracanin M, Hawkins CL, Pattison DI, Davies MJ (2009) Singlet-oxygen-mediated amino acid and protein oxidation: formation of tryptophan peroxides and decomposition products. Free Radic Biol Med 47:92–102. https://doi.org/10.1016/j.freeradbiomed.2009.04.015
Grassi L, Cabrele C (2019) Susceptibility of protein therapeutics to spontaneous chemical modifications by oxidation, cyclization, and elimination reactions. Amino Acids 51:1409–1431. https://doi.org/10.1007/s00726-019-02787-2
Guan Z, Yates NA, Bakhtiar R (2003) Detection and characterization of methionine oxidation in peptides by collision-induced dissociation and electron capture dissociation. J Am Soc Mass Spectrom 14:605–613. https://doi.org/10.1016/S1044-0305(03)00201-0
Gupta V (2013) Glucagon-like peptide-1 analogues: an overview. Indian J Endocrinol Metab 17:413. https://doi.org/10.4103/2230-8210.111625
Hada S, Kim NA, Lim DG, Lim JY, Kim KH, Adhikary P, Jeong SH (2016) Evaluation of antioxidants in protein formulation against oxidative stress using various biophysical methods. Int J Biol Macromol 82:192–200. https://doi.org/10.1016/j.ijbiomac.2015.10.048
Haggag YA (2018) Peptides as drug candidates: limitations and recent development perspectives. Biomed J Sci Tech Res. https://doi.org/10.26717/BJSTR.2018.08.001694
Haile LA, Puig M, Kelley-Baker L, Verthelyi D (2015) Detection of Innate Immune Response Modulating Impurities in Therapeutic Proteins. PLoS ONE 10:e0125078. https://doi.org/10.1371/journal.pone.0125078
Harris AG (1994) Somatostatin and somatostatin analogues: pharmacokinetics and pharmacodynamic effects. Gut 35:S1–S4. https://doi.org/10.1136/gut.35.3_Suppl.S1
Härtl E, Winter G, Besheer A (2013) Influence of Hydroxypropyl-Beta-Cyclodextrin on the Stability of Dilute and Highly Concentrated Immunoglobulin G Formulations. J Pharm Sci 102:4121–4131. https://doi.org/10.1002/jps.23729
Hartmann J, Christel Brand M, Dose K (1981) Formation of specific amino acid sequences during thermal polymerization of amino acids. Biosystems 13:141–147. https://doi.org/10.1016/0303-2647(81)90055-1
Hinterholzer A, Stanojlovic V, Regl C, Huber CG, Cabrele C, Schubert M (2021) Detecting aspartate isomerization and backbone cleavage after aspartate in intact proteins by NMR spectroscopy. J Biomol NMR 75:71–82. https://doi.org/10.1007/s10858-020-00356-4
Hjalte J, Hossain S, Hugerth A, Sjögren H, Wahlgren M, Larsson P, Lundberg D (2022) Aggregation behavior of structurally similar therapeutic peptides investigated by 1H NMR and All-atom molecular dynamics simulations. Mol Pharm 19(3) 904–917. https://doi.org/10.1021/acs.molpharmaceut.1c00883
Huvaere K, Skibsted LH (2009) Light-induced oxidation of tryptophan and histidine. Reactivity of aromatic N -heterocycles toward triplet-excited flavins. J Am Chem Soc 131:8049–8060. https://doi.org/10.1021/ja809039u
Jahn TR, Makin OS, Morris KL, Marshall KE, Tian P, Sikorski P, Serpell LC (2010) The common architecture of cross-β amyloid. J Mol Biol 395:717–727. https://doi.org/10.1016/j.jmb.2009.09.039
Jawa V, Cousens LP, Awwad M, Wakshull E, Kropshofer H, De Groot AS (2013) T-cell dependent immunogenicity of protein therapeutics: preclinical assessment and mitigation. Clin Immunol 149:534–555. https://doi.org/10.1016/j.clim.2013.09.006
Jia L, Sun Y (2017) Protein asparagine deamidation prediction based on structures with machine learning methods. PLoS ONE 12:e0181347. https://doi.org/10.1371/journal.pone.0181347
Jia F, Wang J, Peng J, Zhao P, Kong Z, Wang K, Yan W, Wang R (2017) D-amino acid substitution enhances the stability of antimicrobial peptide polybia-CP. Acta Biochim Biophys Sin (shanghai) 49:916–925. https://doi.org/10.1093/abbs/gmx091
Kang H, Tolbert TJ, Schöneich C (2019) Photoinduced tyrosine side chain fragmentation in IgG4-Fc: mechanisms and solvent isotope effects. Mol Pharm 16:258–272. https://doi.org/10.1021/acs.molpharmaceut.8b00979
Kato K, Nakayoshi T, Kurimoto E, Oda A (2019) Computational studies on the nonenzymatic deamidation mechanisms of glutamine residues. ACS Omega 4:3508–3513. https://doi.org/10.1021/acsomega.8b03199
Kim YH, Berry AH, Spencer DS, Stites WE (2001) Comparing the effect on protein stability of methionine oxidation versus mutagenesis: steps toward engineering oxidative resistance in proteins. Protein Eng Des Sel 14:343–347. https://doi.org/10.1093/protein/14.5.343
Kondejewski LH, Jelokhani-Niaraki M, Farmer SW, Lix B, Kay CM, Sykes BD, Hancock REW, Hodges RS (1999) Dissociation of antimicrobial and hemolytic activities in cyclic peptide diastereomers by systematic alterations in amphipathicity. J Biol Chem 274:13181–13192. https://doi.org/10.1074/jbc.274.19.13181
Kumar KYK, Dama VR, Suchitra C, Maringanti TC (2020) A simple, sensitive, high-resolution, customized, reverse phase ultra-high performance liquid chromatographic method for related substances of a therapeutic peptide (bivalirudin trifluoroacetate) using the quality by design approach. Anal Methods 12:304–316. https://doi.org/10.1039/C9AY01998G
Kuna AK, Ganapathi S, Radha GV (2019) A novel RP- HPLC method development and forced degradation studies for semaglutide in active pharmaceutical ingredients and pharmaceutical dosage form. Int J Res Pharm Sci. https://doi.org/10.26452/ijrps.v10i2.263
Lai X, Tang J, ElSayed MEH (2021) Recent advances in proteolytic stability for peptide, protein, and antibody drug discovery. Expert Opin Drug Discov 16:1467–1482. https://doi.org/10.1080/17460441.2021.1942837
Lajin B, Steiner O, Fasshold L, Zangger K, Goessler W (2019) The identification and chromatographic separation of a new highly analogous impurity of the active pharmaceutical ingredient icatibant. Eur J Pharm Sci 132:121–124. https://doi.org/10.1016/j.ejps.2019.03.003
Lau JL, Dunn MK (2018) Therapeutic peptides: historical perspectives, current development trends, and future directions. Bioorg Med Chem 26:2700–2707. https://doi.org/10.1016/j.bmc.2017.06.052
Lei M, Carcelen T, Walters BT, Zamiri C, Quan C, Hu Y, Nishihara J, Yip H, Woon N, Zhang T, Kao Y-H, Schöneich C (2017) Structure-Based correlation of light-induced histidine reactivity in a model protein. Anal Chem 89:7225–7231. https://doi.org/10.1021/acs.analchem.7b01457
Li B, Borchardt RT, Topp EM, VanderVelde D, Schowen RL (2003) Racemization of an asparagine residue during peptide deamidation. J Am Chem Soc 125:11486–11487. https://doi.org/10.1021/ja0360992
Li X, Lin C, O’Connor PB (2010) Glutamine deamidation: differentiation of glutamic acid and γ-glutamic acid in peptides by electron capture dissociation. Anal Chem 82:3606–3615. https://doi.org/10.1021/ac9028467
Li Y, Clark KA, Tan Z (2018) Methods for engineering therapeutic peptides. Chinese Chem Lett 29:1074–1078. https://doi.org/10.1016/j.cclet.2018.05.027
Li Q, Huang Y, Zhang Y, Cui H, Yin L, Li Y, Yuan A, Hu Y, Wu J (2020) A novel HPLC method for analysis of atosiban and its five related substances in atosiban acetate injection. J Pharm Biomed Anal. https://doi.org/10.1016/j.jpba.2019.112808
Liu F, Lu W, Yin X, Liu J (2016) Mechanistic and kinetic study of singlet O2 oxidation of methionine by on-line electrospray ionization mass spectrometry. J Am Soc Mass Spectrom 27:59–72. https://doi.org/10.1007/s13361-015-1237-4
Liu D, Angelova A, Liu J, Garamus VM, Angelov B, Zhang X, Li Y, Feger G, Li N, Zou A (2019) Self-assembly of mitochondria-specific peptide amphiphiles amplifying lung cancer cell death through targeting the VDAC1–hexokinase-II complex. J Mater Chem B 7:4706–4716. https://doi.org/10.1039/C9TB00629J
Lu J, Xu H, Xia J, Ma J, Xu J, Li Y, Feng J (2020) D- and unnatural amino acid substituted antimicrobial peptides with improved proteolytic resistance and their proteolytic degradation characteristics. Front Microbiol. https://doi.org/10.3389/fmicb.2020.563030
Ma W, Tang C, Lai L (2005) Specificity of trypsin and chymotrypsin: loop-motion-controlled dynamic correlation as a determinant. Biophys J 89:1183–1193. https://doi.org/10.1529/biophysj.104.057158
Mahjoubi N, Fazeli A, Dinarvand R, Khoshayand MR, Shekarchi M, Fazeli MR (2017) Effect of nonionic surfactants (dodecyl maltoside and polysorbate 20) on prevention of aggregation and conformational changes of recombinant human IFNβ_1b induced by light. Iran J Pharm Res IJPR 16:103–111
Mammoser C, Brown B, Dhar S, Rowe L (2017) Analysis of the stability of natural and unnatural amino acids in extraterrestrial conditions. FASEB J. https://doi.org/10.1096/fasebj.31.1_supplement.610.6
Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS (2010) Stability of protein pharmaceuticals: an update. Pharm Res 27:544–575. https://doi.org/10.1007/s11095-009-0045-6
Mant CT, Chen Y, Yan Z, Popa TV, Kovacs JM, Mills JB, Tripet BP, Hodges RS (2007) HPLC analysis and purification of peptides. Peptide characterization and application protocols. Humana Press, Totowa, pp 3–55
MARCUS, F., (2009) Preferential cleavage at aspartyl-prolyl peptide bonds in dilute acid*. Int J Pept Protein Res 25:542–546. https://doi.org/10.1111/j.1399-3011.1985.tb02208.x
Martin S, Mauro D, Sylvia G, Stephen M et al (2019) The benefits of combining UHPLC-UV and MS for peptide impurity profiling. BioPharm Int 32:35–37
McCudden CR, Kraus VB (2006) Biochemistry of amino acid racemization and clinical application to musculoskeletal disease. Clin Biochem 39:1112–1130. https://doi.org/10.1016/j.clinbiochem.2006.07.009
Mensink MA, Frijlink HW, van der Voort Maarschalk K, Hinrichs WLJ (2017) How sugars protect proteins in the solid state and during drying (review): mechanisms of stabilization in relation to stress conditions. Eur J Pharm Biopharm 114:288–295. https://doi.org/10.1016/j.ejpb.2017.01.024
Merutka G, Murphy BM, Payne RW, Wilson GA, Matsuura JE, Henry CS, Manning MC (2016) Stability of lyophilized teriparatide PTH(1-34) after reconstitution. Eur J Pharm Biopharm 99:84–93. https://doi.org/10.1016/j.ejpb.2015.11.012
Meyer JD, Nayar R, Manning MC (2009) Impact of bulking agents on the stability of a lyophilized monoclonal antibody. Eur J Pharm Sci 38:29–38. https://doi.org/10.1016/j.ejps.2009.05.008
Miyamoto T, Takahashi N, Sekine M, Ogawa T, Hidaka M, Homma H, Masaki H (2015) Transition of serine residues to the d-form during the conversion of ovalbumin into heat stable S-ovalbumin. J Pharm Biomed Anal 116:145–149. https://doi.org/10.1016/j.jpba.2015.04.030
Möller MN, Hatch DM, Kim H-YH, Porter NA (2012) Superoxide reaction with tyrosyl radicals generates para -hydroperoxy and para -hydroxy derivatives of tyrosine. J Am Chem Soc 134:16773–16780. https://doi.org/10.1021/ja307215z
Mozziconacci O, Haywood J, Gorman EM, Munson E, Schöneich C (2012) Photolysis of recombinant human insulin in the solid state: formation of a dithiohemiacetal product at the C-Terminal disulfide bond. Pharm Res 29:121–133. https://doi.org/10.1007/s11095-011-0519-1
Muttenthaler M, King GF, Adams DJ, Alewood PF (2021) Trends in peptide drug discovery. Nat Rev Drug Discov 20:309–325. https://doi.org/10.1038/s41573-020-00135-8
Nabuchi Y, Fujiwara E, Ueno K (1995) Oxidation of recombinant human parathyroid hormone: effect of oxidized position on the biological activity. Pharm Res 12:2049–2052. https://doi.org/10.1023/A:1016281031373
Nadler WM, Waidelich D, Kerner A, Hanke S, Berg R, Trumpp A, Rösli C (2017) MALDI versus ESI: the Impact of the ion source on peptide identification. J Proteome Res 16:1207–1215. https://doi.org/10.1021/acs.jproteome.6b00805
Niazi S (2016) Biosimilars and Interchangeable Biologics Tactical Elements, 3rd edn. CRC Press
Nikolaev EN, Popov IA, Kononikhin AS, Indeykina MI, Kukaev EN (2012) High-resolution mass-spectrometry analysis of peptides and proteins. Russ Chem Rev 81:1051–1070. https://doi.org/10.1070/RC2012v081n11ABEH004321
Ohtake S, Kita Y, Arakawa T (2011) Interactions of formulation excipients with proteins in solution and in the dried state. Adv Drug Deliv Rev 63:1053–1073. https://doi.org/10.1016/j.addr.2011.06.011
Ohtake S, Feng S, Shalaev E (2018) Effect of water on the chemical stability of amorphous pharmaceuticals: 2. Deamidation of peptides and proteins. J Pharm Sci 107:42–56. https://doi.org/10.1016/j.xphs.2017.09.003
Oshima H, Kinoshita M (2013) Effects of sugars on the thermal stability of a protein. J Chem Phys 138:245101. https://doi.org/10.1063/1.4811287
Pace AL, Wong RL, Zhang YT, Kao Y-H, Wang YJ (2013) Asparagine Deamidation Dependence on Buffer Type, pH, and Temperature. J Pharm Sci 102:1712–1723. https://doi.org/10.1002/jps.23529
Patel, Jalpa ; Kothari, R., 2011. Stability Considerations for Biopharmaceuticals: Overview of Protein and Peptide Degradation Pathways. Bioprocess Int.
Patel K, Borchardt RT (1990) Chemical pathways of peptide degradation. II. Kinetics of deamidation of an asparaginyl residue in a model hexapeptide. Pharm Res 7:703–711. https://doi.org/10.1023/A:1015807303766
Pedaprolu JN, Bonthu M, Vatchavai B, Kamatham S, Kolli S, Kapuganti AN (2016) A new stability-indicating and validated rp-hplc method for the estimation of liraglutide in bulk and pharmaceutical dosage forms. Eurasian J Anal Chem 12:31–44
Peng IX, Shiea J, Loo RRO, Loo JA (2007) Electrospray-assisted laser desorption/ionization and tandem mass spectrometry of peptides and proteins. Rapid Commun Mass Spectrom 21:2541–2546. https://doi.org/10.1002/rcm.3154
Piszkiewicz D, Landon M, Smith EL (1970) Anomalous cleavage of aspartyl-proline peptide bonds during amino acid sequence determinations. Biochem Biophys Res Commun 40:1173–1178. https://doi.org/10.1016/0006-291X(70)90918-6
Rathore N, Rajan RS (2008) Current Perspectives on stability of protein drug products during formulation, fill and finish operations. Biotechnol Prog 24:504–514. https://doi.org/10.1021/bp070462h
Rehder DS, Borges CR (2010) Cysteine sulfenic acid as an intermediate in disulfide bond formation and nonenzymatic protein folding. Biochemistry 49:7748–7755. https://doi.org/10.1021/bi1008694
Riek R, Eisenberg DS (2016) The activities of amyloids from a structural perspective. Nature 539:227–235. https://doi.org/10.1038/nature20416
Robert J, Sorrieul J, Andrieu A, Mounsef F, Dupoiron D, Devys C (2020) Study of physicochemical stability of ziconotide in medication cassette reservoir. Neuromodulation Technol Neural Interface 23:1034–1041. https://doi.org/10.1111/ner.13218
Ronsein GE, de Oliveira MCB, de Medeiros MHG, Di Mascio P (2011) Mechanism of dioxindolylalanine formation by singlet molecular oxygen-mediated oxidation of tryptophan residues. Photochem Photobiol Sci 10:1727. https://doi.org/10.1039/c1pp05181d
Ryle AP, Sanger F (1955) Disulphide interchange reactions. Biochem J 60:535–540
Sahakijpijarn S, Moon C, Koleng JJ, Williams RO (2019) Formulation composition and process affect counterion for csp7 peptide. Pharmaceutics 11:498. https://doi.org/10.3390/pharmaceutics11100498
Sampson JS, Hawkridge AM, Muddiman DC (2006) Generation and detection of multiply-charged peptides and proteins by matrix-assisted laser desorption electrospray ionization (MALDESI) fourier transform ion cyclotron resonance mass spectrometry. J Am Soc Mass Spectrom 17:1712–1716. https://doi.org/10.1016/j.jasms.2006.08.003
Schoneich C, Zhao F, Madden KP, Bobrowski K (1994) Side chain fragmentation of N-terminal threonine or serine residue induced through intramolecular proton transfer to hydroxy sulfuranyl radical formed at neighboring methionine in dipeptides. J Am Chem Soc 116:4641–4652. https://doi.org/10.1021/ja00090a012
Schöneich C (2005) Methionine oxidation by reactive oxygen species: reaction mechanisms and relevance to Alzheimer’s disease. Biochim Biophys Acta—Proteins Proteomics 1703:111–119. https://doi.org/10.1016/j.bbapap.2004.09.009
Schöneich C (2016) Thiyl radicals and induction of protein degradation. Free Radic Res 50:143–149. https://doi.org/10.3109/10715762.2015.1077385
Schöneich C (2020) Photo-degradation of therapeutic proteins: mechanistic aspects. Pharm Res 37:45. https://doi.org/10.1007/s11095-020-2763-8
Schöneich C (2012) Radical-Based Damage of Sulfur-Containing Amino Acid Residues. In: Smith J (ed) Encyclopedia of radicals in chemistry, biology and materials. John Wiley & Sons Ltd, Chichester UK
Selvin PR (2000) The renaissance of fluorescence resonance energy transfer. Nat Struct Mol Biol. https://doi.org/10.1038/78948
Sestak V, Roh J, Klepalova L, Kovarikova P (2016) A UHPLC-UV-QTOF study on the stability of carfilzomib, a novel proteasome inhibitor. J Pharm Biomed Anal 124:365–373. https://doi.org/10.1016/j.jpba.2016.03.015
Sevilla MD, Yan M, Becker D (1988) Thiol peroxyl radical formation from the reaction of cysteine thiyl radical with molecular oxygen: an ESR investigation. Biochem Biophys Res Commun 155:405–410. https://doi.org/10.1016/S0006-291X(88)81100-8
Shioiri T, Ninomiya K, Yamada S (1972) Diphenylphosphoryl azide. New convenient reagent for a modified curtius reaction and for peptide synthesis. J Am Chem Soc 94:6203–6205. https://doi.org/10.1021/ja00772a052
Sikora K, Jaśkiewicz M, Neubauer D, Migoń D, Kamysz W (2020) The role of counter-ions in peptides—an overview. Pharmaceuticals 13:442. https://doi.org/10.3390/ph13120442
Singh S, Junwal M, Modhe G, Tiwari H, Kurmi M, Parashar N, Sidduri P (2013) Forced degradation studies to assess the stability of drugs and products. TrAC Trends Anal Chem 49:71–88. https://doi.org/10.1016/j.trac.2013.05.006
Sjögren H, Ericsson CA, Evenäs J, Ulvenlund S (2005) Interactions between charged polypeptides and nonionic surfactants. Biophys J 89:4219–4233. https://doi.org/10.1529/biophysj.105.065342
Song Y, Buettner GR (2010) Thermodynamic and kinetic considerations for the reaction of semiquinone radicals to form superoxide and hydrogen peroxide. Free Radic Biol Med 49:919–962. https://doi.org/10.1016/j.freeradbiomed.2010.05.009
Srinivasulu K, Naidu MN, Rajasekhar K, Veerender M, Suryanarayana MV (2012) Development and validation of a stability indicating LC method for the assay and related substances determination of a proteasome inhibitor bortezomib. Chromatogr Res Int 2012:1–13. https://doi.org/10.1155/2012/801720
Stadtman ER (2006) Protein oxidation in aging and age-related diseases. Ann N Y Acad Sci 928:22–38. https://doi.org/10.1111/j.1749-6632.2001.tb05632.x
Steinmann D, Mozziconacci O, Bommana R, Stobaugh JF, Wang YJ, Schöneich C (2017) Photodegradation pathways of protein disulfides: human growth hormone. Pharm Res 34:2756–2778. https://doi.org/10.1007/s11095-017-2256-6
Sun C, Chen L, Shen Z (2019) Mechanisms of gastrointestinal microflora on drug metabolism in clinical practice. Saudi Pharm J 27:1146–1156. https://doi.org/10.1016/j.jsps.2019.09.011
Taddei M (2008) Bicyclic 5–6 Systems with One Bridgehead (Ring Junction) Nitrogen Atom: One Extra Heteroatom 0:1. Comprehensive Heterocyclic Chemistry III. Elsevier, Amsterdam, pp 501–549
Takahashi O, Kirikoshi R (2014) Intramolecular cyclization of aspartic acid residues assisted by three water molecules: a density functional theory study. Comput Sci Discov 7:015005. https://doi.org/10.1088/1749-4699/7/1/015005
Takahashi O, Kirikoshi R, Manabe N (2017) Racemization of serine residues catalyzed by dihydrogen phosphate ion: a computational study. Catalysts 7:363. https://doi.org/10.3390/catal7120363
Takano Y, Oba Y, Furota S, Naraoka H, Ogawa NO, Blattmann TM, Ohkouchi N (2021) Analytical development of seamless procedures on cation-exchange chromatography and ion-pair chromatography with high-precision mass spectrometry for short-chain peptides. Int J Mass Spectrom 463:116529. https://doi.org/10.1016/j.ijms.2021.116529
Tamizi E, Jouyban A (2016) Forced degradation studies of biopharmaceuticals: selection of stress conditions. Eur J Pharm Biopharm 98:26–46. https://doi.org/10.1016/j.ejpb.2015.10.016
Tamizi E, Kenndler E, Jouyban A (2014) A stability indicating capillary electrophoresis method for analysis of buserelin. Iran J Pharm Res IJPR 13:797–807
Tamizi E, Yang Y, Jouyban A, Kelso GF, Boysen RI, Hearn MTW (2016) A capillary electrophoretic–mass spectrometric method for the assessment of octreotide stability under stress conditions. J Chromatogr A 1429:354–363. https://doi.org/10.1016/j.chroma.2015.12.039
Thapa RK, Winther-Larsen HC, Diep DB, Tønnesen HH (2021) Photostability studies of GarKS peptides for topical formulation development. Eur J Pharm Sci 158:105652. https://doi.org/10.1016/j.ejps.2020.105652
Tonie Wright H, Urry DW (1991) Nonenzymatic deamidation of asparaginyl and glutaminyl residues in protein. Crit Rev Biochem Mol Biol 26:1–52. https://doi.org/10.3109/10409239109081719
Trainor K, Broom A, Meiering EM (2017) Exploring the relationships between protein sequence, structure and solubility. Curr Opin Struct Biol 42:136–146. https://doi.org/10.1016/j.sbi.2017.01.004
Trent A, Marullo R, Lin B, Black M, Tirrell M (2011) Structural properties of soluble peptide amphiphile micelles. Soft Matter 7:9572. https://doi.org/10.1039/c1sm05862b
Turell L, Zeida A, Trujillo M (2020) Mechanisms and consequences of protein cysteine oxidation: the role of the initial short-lived intermediates. Essays Biochem 64:55–66. https://doi.org/10.1042/EBC20190053
Ummiti K, Shanmukha Kumar J (2021) Establishment of validated stability indicating purity method based on the stress degradation behavior of gonadotropin-releasing hormone antagonist (ganirelix) in an injectable formulation using HPLC and LC-MS-QTOF. Eur J Mass Spectrom. https://doi.org/10.1177/14690667211005335
Vlasak J, Ionescu R (2011) Fragmentation of Monoclonal Antibodies. Mabs 3:253–263. https://doi.org/10.4161/mabs.3.3.15608
Wakankar AA, Borchardt RT (2006) Formulation considerations for proteins susceptible to asparagine deamidation and aspartate isomerization. J Pharm Sci 95:2321–2336. https://doi.org/10.1002/jps.20740
Wang Z, Rejtar T, Zhou ZS, Karger BL (2010) Desulfurization of cysteine-containing peptides resulting from sample preparation for protein characterization by mass spectrometry. Rapid Commun Mass Spectrom 24:267–275. https://doi.org/10.1002/rcm.4383
Wang Y, Lomakin A, Kanai S, Alex R, Benedek GB (2015) Transformation of oligomers of lipidated peptide induced by change in pH. Mol Pharm 12:411–419. https://doi.org/10.1021/mp500519s
Wang L, Wang N, Zhang W, Cheng X, Yan Z, Shao G, Wang X, Wang R, Fu C (2022) Therapeutic peptides: current applications and future directions. Signal Transduct Target Ther 7:48. https://doi.org/10.1038/s41392-022-00904-4
Windisch V, Deluccia F, Duhau L, Herman F, Mencel JJ, Tang S-Y, Vuilhorgne M (1997) Degradation pathways of salmon calcitonin in aqueous solution. J Pharm Sci 86:359–364. https://doi.org/10.1021/js9602305
Witt M, Fuchser J, Baykut G (2003) Fourier transform ion cyclotron resonance mass spectrometry with NanoLC/microelectrospray ionization and matrix-assisted laser desorption/ionization: Analytical performance in peptide mass fingerprint analysis. J Am Soc Mass Spectrom 14:553–561. https://doi.org/10.1016/S1044-0305(03)00138-7
Wright A, Bubb WA, Hawkins CL, Davies MJ (2007) Singlet oxygen-mediated protein oxidation: evidence for the formation of reactive side chain peroxides on tyrosine residues. Photochem Photobiol 76:35–46. https://doi.org/10.1562/0031-8655(2002)0760035SOMPOE2.0.CO2
Wu LC, Chen F, Lee SL, Raw A, Yu LX (2020) Building parity between brand and generic peptide products : regulatory and scienti fi c considerations for quality of synthetic peptides. Int J Pharm 518:320–334. https://doi.org/10.1016/j.ijpharm.2016.12.051
Yang Y (2016) β-Elimination Side Reactions. Side Reactions in Peptide Synthesis. Elsevier, Amsterdam, pp 33–42
Yang H, Zubarev RA (2010) Mass spectrometric analysis of asparagine deamidation and aspartate isomerization in polypeptides. Electrophoresis 31:1764–1772. https://doi.org/10.1002/elps.201000027
Yashiro H, White RC, Yurkovskaya AV, Forbes MDE (2005) Methionine radical cation: structural studies as a function of pH using X- and Q-band time-resolved electron paramagnetic resonance spectroscopy. J Phys Chem A 109:5855–5864. https://doi.org/10.1021/jp051551k
Yazıcı, E.Y., Deveci, H., 2010. Factors Affecting Decomposition of Hydrogen Peroxide. https://doi.org/10.13140/RG.2.1.1530.0648
Yin L, Huang Y, Wang C, Dong H, Ding Y, Huang B, Wu J (2021) Development of a novel and stability indicating RP-HPLC-UV method for simultaneous analysis of carbetocin and ten impurities in carbetocin injection products. Chromatographia 84:967–978. https://doi.org/10.1007/s10337-021-04083-2
Yuan Y, Li Y-B, Tai Z-F, Xie Y-P, Pu X-F, Gao J (2018) Study of forced degradation behavior of pramlintide acetate by HPLC and LC–MS. J Food Drug Anal 26:409–415. https://doi.org/10.1016/j.jfda.2017.07.009
Zapadka KL, Becher FJ, Gomes dos Santos AL, Jackson SE (2017) Factors affecting the physical stability (aggregation) of peptide therapeutics. Interface Focus 7:20170030. https://doi.org/10.1098/rsfs.2017.0030
Zbacnik TJ, Holcomb RE, Katayama DS, Murphy BM, Payne RW, Coccaro RC, Evans GJ, Matsuura JE, Henry CS, Manning MC (2017) Role of buffers in protein formulations. J Pharm Sci 106:713–733. https://doi.org/10.1016/j.xphs.2016.11.014
Zeng K, Geerlof-Vidavisky I, Gucinski A, Jiang X, Boyne MT (2015) Liquid chromatography-high resolution mass spectrometry for peptide drug quality control. AAPS J 17:643–651. https://doi.org/10.1208/s12248-015-9730-z
Zhang J, Kalonia DS (2007) The effect of neighboring amino acid residues and solution environment on the oxidative stability of tyrosine in small peptides. AAPS PharmSciTech 8:176. https://doi.org/10.1208/pt0804102
Zhang F, Angelova A, Garamus VM, Angelov B, Tu S, Kong L, Zhang X, Li N, Zou A (2021a) Mitochondrial voltage-dependent anion channel 1–hexokinase-II complex-targeted strategy for melanoma inhibition using designed multiblock peptide amphiphiles. ACS Appl Mater Interfaces 13:35281–35293. https://doi.org/10.1021/acsami.1c04385
Zhang X, Belousoff MJ, Liang Y-L, Danev R, Sexton PM, Wootten D (2021b) Structure and dynamics of semaglutide- and taspoglutide-bound GLP-1R-Gs complexes. Cell Rep 36:109374. https://doi.org/10.1016/j.celrep.2021.109374
Acknowledgements
The authors would like to thank Institute of Pharmacy, Nirma University, Ahmedabad, India for providing the necessary facilities.
Author information
Authors and Affiliations
Contributions
1—Conceptualization; Visualization, Investigation, Writing the manuscript. 2—Conceptualization, Writing the manuscript. 3 - Conceptualization, Visualization, Validation, Supervision, Writing—review.
Corresponding author
Ethics declarations
Competing interests
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Patel, S., Vyas, V.K. & Mehta, P.J. A Review on Forced Degradation Strategies to Establish the Stability of Therapeutic Peptide Formulations. Int J Pept Res Ther 29, 22 (2023). https://doi.org/10.1007/s10989-023-10492-8
Accepted:
Published:
DOI: https://doi.org/10.1007/s10989-023-10492-8