Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis

García-Artalejo, Judey Aymed; Mancera-Arteu, Montserrat; Sanz‐Nebot, Victoria; Rodríguez, Teresita; Giménez, Estela

doi:10.1016/j.jpba.2022.114686

Cited by 3 publications

(8 citation statements)

References 28 publications

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“…EPOCIM is a biosimilar commercialized for the treatment of anemias, and its production may result in a slightly different glycosylation pattern from that of EPO-CRS. NeuroEPO plus is a basic rhEPO under investigation for the treatment of neurodegenerative diseases Table shows the glycoforms of the O 126 and N 83 glycopeptides of EPOCIM and NeuroEPO plus detected by PBA-SPE-CE-MS at 50 mg·L –1 digested protein.…”

Section: Resultsmentioning

confidence: 99%

“…NeuroEPO plus is a basic rhEPO under investigation for the treatment of neurodegenerative diseases. 4 Table 3 shows the glycoforms of the O 126 and N 83 glycopeptides of EPOCIM and NeuroEPO plus detected by PBA-SPE-CE-MS at 50 mg·L –1 digested protein. NeuroEPO plus showed a superior amount of less sialylated glycoforms in both O 126 and N 83 glycopeptides, while the glycoform composition of EPOCIM was similar to that of EPO-CRS (compare Tables 2 and 3 for N 83 glycopeptide).…”

Section: Resultsmentioning

confidence: 99%

Section: Application To Rhepo Biosimilarsmentioning

confidence: 99%

“…However, it is still necessary to develop novel analytical platforms based on mass spectrometry (MS) not only to improve the quality control of the existing rhEPO biosimilars but also to deeply characterize those products that are under investigation for other clinical applications. This is the case of NeuroEPO plus, a recently developed rhEPO with a low sialic acid content that is currently in phase II–III clinical trials in Parkinson’s and Alzheimer’s diseases. , Among the different MS-based strategies to analyze protein glycosylation, the bottom-up analysis of the glycopeptides obtained after enzymatic digestion of the target glycoprotein offers important advantages. Indeed, it provides information not only about the glycan structures but also about the amino acids to which they are attached and hence about the glycosites of the carrier protein …”

Section: Introductionmentioning

confidence: 99%

“…This is the case of NeuroEPO plus, a recently developed rhEPO with a low sialic acid content that is currently in phase II–III clinical trials in Parkinson’s and Alzheimer’s diseases. 3 , 4 Among the different MS-based strategies to analyze protein glycosylation, the bottom-up analysis of the glycopeptides obtained after enzymatic digestion of the target glycoprotein offers important advantages. Indeed, it provides information not only about the glycan structures but also about the amino acids to which they are attached and hence about the glycosites of the carrier protein.…”

Section: Introductionmentioning

confidence: 99%

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Sensitive Analysis of Recombinant Human Erythropoietin Glycopeptides by On-Line Phenylboronic Acid Solid-Phase Extraction Capillary Electrophoresis Mass Spectrometry

et al. 2023

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In this study, several chromatographic sorbents: porous graphitic carbon (PGC), aminopropyl hydrophilic interaction (aminopropyl-HILIC), and phenylboronic acid (PBA) were assessed for the analysis of glycopeptides by on-line solid-phase extraction capillary electrophoresis mass spectrometry (SPE-CE-MS). As the PBA sorbent provided the most promising results, a PBA-SPE-CE-MS method was developed for the selective and sensitive preconcentration of glycopeptides from enzymatic digests of glycoproteins. Recombinant human erythropoietin (rhEPO) was selected as the model glycoprotein and subjected to enzymatic digestion with several proteases. The tryptic O126 and N83 glycopeptides from rhEPO were targeted to optimize the methodology. Under the optimized conditions, intraday precision, linearity, limits of detection (LODs), and microcartridge lifetime were evaluated, obtaining improved results compared to that from a previously reported TiO2-SPE-CE-MS method, especially for LODs of N-glycopeptides (up to 500 times lower than by CE-MS and up to 200 times lower than by TiO2-SPE-CE-MS). Moreover, rhEPO Glu-C digests were also analyzed by PBA-SPE-CE-MS to better characterize N24 and N38 glycopeptides. Finally, the established method was used to analyze two rhEPO products (EPOCIM and NeuroEPO plus), demonstrating its applicability in biopharmaceutical analysis. The sensitivity of the proposed PBA-SPE-CE-MS method improves the existing CE-MS methodologies for glycopeptide analysis and shows a great potential in glycoprotein analysis to deeply characterize protein glycosites even at low concentrations of the protein digest.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Application To Rhepo Biosimilarsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Sensitive Analysis of Recombinant Human Erythropoietin Glycopeptides by On-Line Phenylboronic Acid Solid-Phase Extraction Capillary Electrophoresis Mass Spectrometry

et al. 2023

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NeuroEPO plus (NeuralCIM®) in mild-to-moderate Alzheimer’s clinical syndrome: the ATHENEA randomized clinical trial

Sosa,

Bringas,

Urrutia

et al. 2023

Alz Res Therapy

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Background NeuroEPO plus is a recombinant human erythropoietin without erythropoietic activity and shorter plasma half-life due to its low sialic acid content. NeuroEPO plus prevents oxidative damage, neuroinflammation, apoptosis and cognitive deficit in an Alzheimer’s disease (AD) models. The aim of this study was to assess efficacy and safety of neuroEPO plus. Methods This was a double-blind, randomized, placebo-controlled, phase 2–3 trial involving participants ≥ 50 years of age with mild-to-moderate AD clinical syndrome. Participants were randomized in a 1:1:1 ratio to receive 0.5 or 1.0 mg of neuroEPO plus or placebo intranasally 3 times/week for 48 weeks. The primary outcome was change in the 11-item cognitive subscale of the AD Assessment Scale (ADAS-Cog11) score from baseline to 48 weeks (range, 0 to 70; higher scores indicate greater impairment). Secondary outcomes included CIBIC+, GDS, MoCA, NPI, Activities of Daily Living Scales, cerebral perfusion, and hippocampal volume. Results A total of 174 participants were enrolled and 170 were treated (57 in neuroEPO plus 0.5 mg, 56 in neuroEPO plus 1.0 mg and 57 in placebo group). Mean age, 74.0 years; 121 (71.2%) women and 85% completed the trial. The median change in ADAS-Cog11 score at 48 weeks was −3.0 (95% CI, −4.3 to −1.7) in the 0.5 mg neuroEPO plus group, −4.0 (95% CI, −5.9 to −2.1) in the 1.0 mg neuroEPO plus group and 4.0 (95% CI, 1.9 to 6.1) in the placebo group. The difference of neuroEPO plus 0.5 mg vs. placebo was 7.0 points (95% CI, 4.5–9.5) P = 0.000 and between the neuroEPO plus 1.0 mg vs. placebo was 8.0 points (95% CI, 5.2–10.8) P = 0.000. NeuroEPO plus treatment induced a statistically significant improvement in some of clinical secondary outcomes vs. placebo including CIBIC+, GDS, MoCA, NPI, and the brain perfusion. Conclusions Among participants with mild-moderate Alzheimer’s disease clinical syndrome, neuroEPO plus improved the cognitive evaluation at 48 weeks, with a very good safety profile. Larger trials are warranted to determine the efficacy and safety of neuroEPO plus in Alzheimer’s disease. Trial registration https://rpcec.sld.cu Identifier: RPCEC00000232.

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Evidence of the neuroprotective potential of NeuroEPO in preclinical studies and clinical trials: A systematic review

Fernández,

Pérez,

Román

et al. 2023

Preprint

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Introduction Neuroprotection is an essential purpose in the treatment of neurological disorders and several investigations have evaluated the usefulness of NeuroEPO in preclinical studies and clinical trials. Objective To evaluate the neuroprotective potential of NeuroEPO in neurological disorders. Material and Methods A systematic review of original articles in Spanish and English was conducted using PRISMA methodology, guided by the question: Is NeuroEPO a safe and effective therapeutic candidate for neuroprotection in neurological disorders? LILACS and PubMed databases, and Google Scholar search engine were used. Results With the strategy followed, a total of 20 articles were included for synthesis and review. Preclinical studies were conducted in biomodels of cerebral ischemia and Alzheimer's disease, while clinical trials were conducted in patients with neurodegenerative diseases. Safety, tolerability, and efficacy of NeuroEPO were found when administered intranasally with varying doses, frequency of administration, and duration of intervention. Neuroprotective effects were observed in all studies, and some molecular mechanisms mediating these effects were evidenced. Conclusions NeuroEPO has been reported to be a safe product, without hematopoietic effects, and well tolerated intranasally, which provides benefits in conditions of ischemic or degenerative brain damage by stimulating endogenous neuroprotection mechanisms with antioxidant, anti-inflammatory, and antiapoptotic action. The results support the continuity of studies aimed at enriching the scientific evidence of the potential of NeuroEPO for the treatment of neurological disorders.

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Characterizing a novel hyposialylated erythropoietin by intact glycoprotein and glycan analysis

Cited by 3 publications

References 28 publications

Sensitive Analysis of Recombinant Human Erythropoietin Glycopeptides by On-Line Phenylboronic Acid Solid-Phase Extraction Capillary Electrophoresis Mass Spectrometry

Sensitive Analysis of Recombinant Human Erythropoietin Glycopeptides by On-Line Phenylboronic Acid Solid-Phase Extraction Capillary Electrophoresis Mass Spectrometry

NeuroEPO plus (NeuralCIM®) in mild-to-moderate Alzheimer’s clinical syndrome: the ATHENEA randomized clinical trial

Evidence of the neuroprotective potential of NeuroEPO in preclinical studies and clinical trials: A systematic review

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