DoE approach for development of localized controlled release microspheres of Vancomycin for treatment of septic arthritis

Background Septic arthritis is a worse condition of RA that is associated with significant morbidity and mortality. Septic arthritis develops due to direct introduction or invasion of pathogens. The objective of the present study was to formulate Vancomycin hydrochloride-loaded microspheres (VMS) based on Box–Behnken design (BBD) and evaluate its efficacy against septic arthritis. The intraarticular administration of optimized Vancomycin hydrochloride-loaded microspheres (OVMS) can reduce dose size, dosing frequency and systemic exposure with local targeted delivery. Results OVMS was further characterized for its drug–polymer compatibility using differential scanning calorimetry and Fourier transmission infrared spectroscopy. In vitro antibacterial activity was determined using the cup–plate method and in vivo anti-arthritic efficacy was evaluated by gross examination of septic arthritis. DSC and FTIR studies exhibited no interaction or incompatibilities between the drug and polymer. SEM images revealed that OVMS were spherical. It followed the first-order release rate according to Fick's law. The micromeritic properties indicated good flow property of OVMS. The zone of inhibition by OVMS was 1.5 cm against S. aureus. In vivo antibacterial study revealed that OVMS was significant in reducing septic arthritis and bacterial load, i.e., 110.1 CFU/ml in comparison with the control group (850 CFU/ml). Conclusions Thus, OVMS may be used as an effective formulation for the treatment of septic arthritis as compared to marketed IV vancomycin injection after clinical studies.

Liposomes as Vancomycin Hydrochloride Delivery System

Liposomes are being used as unique drug delivery systems due to their ability to encapsulate both hydrophilic and hydrophobic drugs, as well as for the fact that they improve the disadvantages of free drug administration. However, liposomes have a significant disadvantage - low encapsulation efficiency. In the research carried out, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol (Chol), in the ratio (n/n) of 2:1, 3:1 and 4:1 respectively, were used to prepare the liposomes. Blank liposomes (LIP) and vancomycin hydrochloride (VANKA) containing liposomes (VANKA-LIP) were prepared for each of the DSPC and Chol compositions. The aim of our study was to evaluate the effect of liposome composition on the VANKA encapsulation efficiency and release kinetics.

Determination of Vancomycin B and Vancomycin Impurities by Liquid Chromatography

The preferred test methods for control of product-related impurities in medicinal products are high-performance liquid chromatography (HPLC) with a fine sorbent, and ultra-performance liquid chromatography (UPLC), which allow for better chromatographic separation of active substances and related impurities, reduction of time costs, and saving of material resources. The aim of the study was to develop HPLC and UPLC test procedures and assess the chromatographic separation capacity and efficiency in order to improve determination of the main vancomycin component and related impurities. Materials and methods: vancomycin hydrochloride lyophilisate for oral solution and solution for injection, and vancomycin hydrochloride reference standard (USP RS) were used as test objects. Agilent 1290 Infinity liquid chromatography system, and Chromolith® Performance RP-18e, Kinetex C18, Nucleodur C18 Isis, Zorbax RRHD Eclipse Plus C18, and LiChrospher® RP-18 columns were used for the testing. Results: HPLC analysis using a Chromolith® column (100×4.6 mm) reduces the testing time by 10 minutes compared to the USP test procedure, and by 15 minutes compared to the British Pharmacopoeia procedure. The proposed test procedure requires less eluent and increases chromatographic separation efficiency. UPLC analysis using a Kinetex C18 column (50×4.6 mm, 2.6 μm) made it possible to reduce the testing time by two thirds compared to the British Pharmacopoeia procedure. The use of isocratic elution greatly simplified the testing. The testing time under the proposed chromatographic conditions was 10 minutes. Conclusions: the selected HPLC and UPLC test conditions made it possible to significantly reduce the time of testing, minimise the use of expensive reagents, and increase efficiency of chromatographic separation in the determination of vancomycin impurities and the main component Vancomycin B.

Spectral Data Analysis and Identification of Vancomycin Hydrochloride

Objective: To establish a method for the determination of the chemical structure of vancomycin hydrochloride.Methods: Nuclear magnetic resonance spectroscopy and mass spectrometry were conducted to analyze the chemical structure of vancomycin hydrochloride.Results: In this study, the target compound (1) was identified as (Sα)-(3S, 6R, 7R, 22R, 23S, 26S, 36R, 38αR)-44-[[2-O-(3-amino-2, 3, 6-trideoxy-3-C-methyl-α-L-lyso-hexopyranosyl)-β-D-glucopyranosyl] oxy]-3-(carbamoylmethyl)-10, 19-dichloro-7, 22, 28, 30, 32-pentahydroxy-6-[[(2R)-4-methyl-2-(methylamino) pentanoyl] amino]-2, 5, 24, 38, 39-pentaoxo-2, 3, 4, 5, 6, 7, 23, 24, 25, 26, 36, 37, 38, 38α-tetradecahydro-22H-8, 11: 18, 21-dietheno-23, 36-(iminomethano)-13, 16: 31, 35-dimetheno-1H, 13H-[1, 6, 9] oxadiazacyclohexadecino [4, 5-m] [10, 2, 16]-benzoxadiazacyclotetracosine-26-carboxylic acid hydrochloride.Conclusion: The method used in this study is accurate and can be used for the production and structural elucidation of vancomycin hydrochloride.