ABSTRACT
Pegfilgrastim is administered as an adjunct to chemotherapy to reduce the incidence of febrile neutropenia and associated infectious complications. Lupin's Pegfilgrastim is a proposed biosimilar to the U.S.-referenced Neulasta®. Demonstration of biosimilarity requires extensive physicochemical and functional characterization of the biosimilar, and demonstration of analytical similarity to the reference product, in addition to clinical studies. This work is a case study for demonstrating the analytical similarity of Armlupeg (Lupin's Pegfilgrastim) to Neulasta® with respect to structural and physicochemical attributes using several robust, orthogonal, and state-of-the-art techniques including high-end liquid chromatography, mass spectrometry, and spectroscopy techniques; circular dichroism; differential scanning calorimetry; nuclear magnetic resonance; analytical ultracentrifugation; and micro-flow imaging. Functional similarity was demonstrated using an in vitro cell proliferation assay to measure relative potency and surface plasmon resonance to measure receptor binding kinetics. Furthermore, comparative forced-degradation studies were performed to study the degradation of the products under stress conditions. The product attributes were ranked based on a critical quality attributes risk score according to their potential clinical impact. Based on criticality, all analyses were statistically evaluated to conclude analytical similarity. Lupin's Pegfilgrastim was comparable to Neulasta® as demonstrated via structural, functional, and purity analyses. Lupin's Pegfilgrastim complied with the quality and statistical ranges established using Neulasta®. Both products follow the same degradation pathways under stress conditions as observed in the forced-degradation studies. No new impurity or degradation product was observed in Lupin's Pegfilgrastim. These data conclusively demonstrate the analytical similarity of Lupin's Pegfilgrastim and Neulasta®.
Subject(s)
Biosimilar Pharmaceuticals , Biosimilar Pharmaceuticals/therapeutic use , Filgrastim , Polyethylene Glycols/chemistry , Research DesignABSTRACT
The predisposing factors of invasive fungal disease in COVID 19 infection are still debatable because of the limited human understanding of the virus with the current literature. In this study, we have tried to correlate the various predisposing factors influencing the clinical profile and treatment outcomes in patients with covid associated mucormycosis (CAM). It is a retrospective analysis of cases of CAM during the second wave of COVID 19 infection, which was managed in the department of Otorhinolaryngology from Dec 1, 2020, to June 10, 2021. The detailed clinical, radiological and management of patients with CAM were collected, recorded, evaluated and correlated with the predisposing factors. Of the total, 46 patients, 44(95.65%) were diabetic and 41 patients had a previous history of steroid intake. When clinical parameters were compared between blood sugar < 200 mg/dl and > 200 mg/dl, the old and newly diagnosed diabetes mellitus in patients with CAM, there was no significant differences in any of the above clinical parameters (p > 0.05), except the hospital stay (p = 0,004). Steroid intake in patients with coexisting DM associated with CAM is considered the most important factor for the development of the CAM. There was are no significant difference in any of the clinical/treatment outcomes in patients with CAM with respect to the initial blood sugar, except for the hospital stay. A large sample size with a long-term follow-up period may be needed for a better understanding of common predisposing factors for the development of CAM.
ABSTRACT
Reteplase (rPA) is a thrombolytic agent used for the treatment of acute myocardial infarction. We studied the expression of rPA and its selected asparagine mutants after integration into the Pichia genome. Though methanol induction of the native and the rPA mutants showed similar expression levels (~200-250 mg/L), the mutants displayed significant loss of protease activity. Strikingly, the clot lysis activities of these mutants were considerably different. While mutation of Asn(12) (N12P) of the Kringle 2 domain showed delayed clot lysis activity (t(1/2) = 38 min) compared to the native rPA (t(1/2) = 33 min), a faster rate of clot lysis (t(1/2) = 27 min) was observed when the Asn(278) (N278S) of the serine protease domain was mutated. Interestingly, the slowest clot lysis activity (t(1/2) = 49 min) demonstrated by the double mutant (N12P, N278S) suggests the dominant role of Asn(12) in regulating the fibrinolytic activity of rPA. The results presented in this paper indicate that the fibrinolytic and the proteolytic activities of rPA are independent of each other.
