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@#Objective To develop quality control methods for the key quality properties of recombinant monoclonal antibodies against proprotein convertase subtilisin/Kexin 9(PCSK9). Methods According to the corresponding requirements of Chinese Pharmacopoeia(Volume Ⅲ,2020 edition)and International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH),the PCSK9 monoclonal antibody was identified for the identity by reducing trypsin peptide fingerprint mapping analysis,identified for the specificity by ELISA,determined for the charge heterogeneity by imaging capillary isoelectric focusing electrophoresis(icIEF),detected for the variant purity by reducing/non-reducing capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS),measured for the content of monomers,high molecular weight substances and low molecular weight substances by size-exclusion chromatography high performance liquid chromatography(SEC-HPLC),analyzed for the glycotype with N-glycan labeled with 2AB after carbohydrate cutting by Waters ACQUITY UPLC system with fluorescence detector,determined for the biological potency with HepG2 cell line and low-density lipoprotein(LDL),and detected for the content of polysorbate 20 by the liquid phase detection system(CAD detector). Results PCSK9 monoclonal antibody sample had characteristic peptide segments,and the peptide fingerprint was consistent with that of reference substance. The sample contained specific antibodies;The relative area percentages of main peak,acid peak and alkaline peak of the sample were(49. 27 ± 0. 38)%,(46. 44 ± 0. 35)% and(4. 28 ± 0. 04)%,respectively. The relative area percentages of“heavy chain + light chain”peak,non-glycosyl main peak and low molecular weight substance peak were(94. 16 ± 0. 82)%,(4. 11 ± 0. 76)% and(0. 85 ± 0. 20)%,respectively. The relative area percentages of main peak,nonglycosyl main peak and low molecular weight substance peak were(94. 27 ± 0. 22)%,(2. 85 ± 0. 08)% and(2. 88 ± 0. 15)%,respectively. The relative area percentages of monomer,high molecular weight substance and low molecular weight substance were(98. 30 ± 0. 03)%,(0. 75 ± 0. 01)% and(0. 96 ± 0. 02)%,respectively. The relative percentages of G0F,G1F,G2F and non-fucosylated(G0 + G1 + G2 + Man5)were(39. 31 ± 0. 54)%,(34. 69 ± 0. 41)%,(9. 09 ± 0. 14)% and(12. 61 ± 0. 50)%,respectively. The relative biological potency was(101. 64 ± 3. 61)% of the reference. The content of polysorbate 20 was(0. 012 ± 0. 000 3)%. Conclusion According to the key quality attribute of monoclonal antibody against PCSK9,the corresponding key quality control methods have been established,which can ensure the safety,effectiveness and quality controllability of the product.
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Resumen: La hiperlipidemia es altamente prevalente y contribuye de forma sustancial a la enfermedad cardiovascular aterosclerótica, que es una de las principales causas de morbilidad y mortalidad en Colombia. La reducción del colesterol LDL (c-LDL) produce una disminución del riesgo de enfermedad cardiovascular aterosclerótica y de eventos cardiovasculares adversos. La terapia dirigida a la proproteína convertasa subtilisina/kexina tipo 9 (PCSK9; su sigla en inglés) ha surgido como una herramienta novedosa para el tratamiento de la hiperlipidemia. Inclisiran es un ARN pequeño de doble hebra, que actúa inhibiendo la transcripción de PCSK-9 en los hepatocitos, lo que conduce a una reducción marcada y sostenida del c-LDL. En contraste con otras terapias hipolipemiantes, como estatinas, ezetimibe y anticuerpos monoclonales (MAbs; su sigla en inglés) e inhibidores de PCSK9, inclisiran propone un régimen de dosificación infrecuente de dos o tres veces al año. Su efecto prolongado representa una ventaja frente al incumplimiento del tratamiento, que es una de las principales causas por las que no se alcanzan los objetivos de c-LDL con la terapia estándar. Esta revisión tiene como objetivo presentar y discutir los datos científicos actuales con relación a la eficacia, tolerabilidad y seguridad del inclisiran en el tratamiento de la hipercolesterolemia.
Abstract: Hyperlipidemia is a highly prevalent condition and contributes substantially to atherosclerotic cardiovascular disease (ASCVD), which is one of the main causes of morbidity and mortality in Colombia. The reduction of LDL cholesterol (LDL-C) decreases the risk of ASCVD and adverse cardiovascular events. Targeted therapy for the proprotein convertase subtilisin/kexin type 9 (PCSK-9) has emerged as a novel tool for the treatment of hyperlipidemia. Inclisiran is a small double-stranded small interfering RNA that acts by blocking PCSK-9 transcription in hepatocytes, leading to a marked and sustained reduction in circulating LDL-C levels. In contrast to other lipid-lowering therapies such as statins, ezetimibe and monoclonal antibodies (MAbs) PCSK-9 inhibitors, Inclisiran proposes an infrequent dosing regimen of twice or three times a year. Its prolonged effect represents an advantage over non-compliance of the treatment, which is one of the main reasons why LDL-C goals are not achieved with standard therapy. This review aims to present and discuss current scientific data regarding the efficacy, tolerability and safety of Inclisiran in the treatment of hypercholesterolemia.
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Proprotein convertase subtilisin/kexin 9 (PCSK9) inhibitor is recently added to the list of effective lipid-lowering drugs in addition to statins, which can reduce low density lipoprotein cholesterol, treat increased low density lipoprotein cholesterol (LDL-C) and atherosclerotic cardiovascular disease (ASCVD) patients. In the past 30 years, the prevalence of dyslipidemia increased significantly. LDL-C characterized dyslipidemia is an important risk factor of ASCVD and a part of metabolic syndrome, which exists before or after or at the same time of the appearance of obesity, diabetes, and glucose intolerance, coronary heart disease and other diseases. Decreasing LDL-C level can significantly reduce the incidence and mortality risk of ASCVD. In this paper, the research on the regulation of dyslipidemia by PCSK9 inhibitors is discussed.
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Objective To assess the expression and significance of proprotein convertase subtilisin kexin 9 (PCSK9) in patients with rheumatoid arthritis (RA).Methods Sixty-five RA patients and forty-seven healthy controls were recruited in this study.The body mass index (BMI) and serum total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL),lipoprotein a,low density lipoprotein (LDL),very low density lipoprotein(VLDL),apolipoprotein A (ApoA),apolipoprotein B (ApoB) and the ratio of LDL-C/HDL-C were tested.Other parameters included disease activity score 28 (DAS28),rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP) antibody,erythrocyte sedimentation rate (ESR),c-reactive protein (CRP).Serum PCSK9 level was measured by ELISA and compared between RA patients and healthy controls.Results (1) The serum PCSK9 levels in RA patients were higher than those in healthy controls [(409.36 ±223.52) μg/L vs (292.19 ± 109.79) μg/L,P ＜ 0.05].(2) Compared with subgroup of moderate and low active disease and patients in remission,PCSK9 was significantly higher in patients with highly active disease (P ＜ 0.05).(3) The serum PCSK9 levels were positively correlated with RF,TC,TG,LDL,very low density lipoprotein (VLDL),ApoB,with r values as 0.303,0.490,0.320,0.451,0.319,0.463,respectively (P ＜ 0.05).(4) Multiple stepwise regression analysis showed that DAS28,RF,TC and LDL-C/HDL-C were relevant factors for PCSK9 in RA patients.Conclusions The serum PCSK9 level is elevated in RA patients,which is related to RF,disease activity,TC,TG,LDL,VLDL,ApoB.This suggests that PCSK9 is potentially linked to inflammatory reaction and lipid metabolism in rheumatoid arthritis.