Polymer selection for amorphous solid dispersion of a new drug candidate by investigation of drug polymer molecular interactions.

The antitumor drug candidate X-05 is being developed as an innovative anti-lung cancer drug candidate due to its excellent antitumour activity. A Caco-2 cell permeability study and solubility study confirmed that X-05 belonged to BCS class or compounds. Therefore, the main challenge is to develop appropriate preparations for preclinical studies and further clinical phase research. By evaluating the preliminary results of kinetic solubility in biorelevant media and the structural analysis of X-05 and polymers, three polymers PVP K30, PVP VA 64 and HPMCAS, which may have intermolecular interactions with X-05, were chosen to select the optimal carrier for X-05 to prepare amorphous solid dispersions (ASDs). ASD X-05-PVP VA 64 was selected as the optimal polymer by evaluating its kinetic solubility in biorelevant media and solid stability. The physical and chemical properties of ASD X-05-PVP VA 64 remain stable when the drug loading is as high as 50%. The drug-polymer interactions of ASD X-05-PVP VA 64 were studied by ultraviolet spectrophotometry, nuclear magnetic resonance spectrometry, infrared and Raman spectrophotometry, and the results indicated that the intermolecular hydrogen bond interaction between the drug and polymer was the foundation of the solubilization and stabilization of X-05 in PVP VA 64.

A Review of Liposomes as a Drug Delivery System: Current Status of Approved Products, Regulatory Environments, and Future Perspectives.

Liposomes have been considered promising and versatile drug vesicles. Compared with traditional drug delivery systems, liposomes exhibit better properties, including site-targeting, sustained or controlled release, protection of drugs from degradation and clearance, superior therapeutic effects, and lower toxic side effects. Given these merits, several liposomal drug products have been successfully approved and used in clinics over the last couple of decades. In this review, the liposomal drug products approved by the U.S. Food and Drug Administration (FDA) and European Medicines Agency (EMA) are discussed. Based on the published approval package in the FDA and European public assessment report (EPAR) in EMA, the critical chemistry information and mature pharmaceutical technologies applied in the marketed liposomal products, including the lipid excipient, manufacturing methods, nanosizing technique, drug loading methods, as well as critical quality attributions (CQAs) of products, are introduced. Additionally, the current regulatory guidance and future perspectives related to liposomal products are summarized. This knowledge can be used for research and development of the liposomal drug candidates under various pipelines, including the laboratory bench, pilot plant, and commercial manufacturing.

Comparative anti-inflammatory effect of curcumin at air-liquid interface and submerged conditions using lipopolysaccharide stimulated human lung epithelial A549 cells.

BACKGROUND: Inhalation of aerosolized drugs is a promising entry route for rapid and non-invasive therapeutics delivery to the lung. Curcumin exhibits potent anti-inflammatory properties, which are effective for use in lung diseases. The anti-inflammatory properties of curcumin have been widely studied in vitro with cells cultured in submerged conditions, however, the effectiveness using air-liquid interface (ALI) exposure is currently unknown. METHODS: The anti-inflammatory effect of curcumin under both ALI and submerged conditions was investigated in the present study. Lipopolysaccharide (LPS) stimulated A549 cells were exposed to curcumin under ALI (10-100 µM) using a dose-controlled air-liquid interface cell exposure (ALICE)-CLOUD system and submerged cell culture conditions (1-20 µM). The expression of pro-inflammatory cytokines (interleukin (IL)-6, IL-8), cell viability and cytotoxicity were studied for each exposure scenario. The cellular uptake behaviour of curcumin was studied with an equipotent cell-based dose (200 pmol/106 cell) at various time points up to 24 h. RESULTS: The ALI delivery profile proved to be rapid, efficient and reproducible. For the doses studied, no significant effect on cell viability and cytotoxicity were observed. ALI exposure of curcumin was more effective in reducing pro-inflammatory cytokines expression in lung epithelial cells compared with submerged cell cultures. Furthermore, rapid cellular uptake and higher intracellular doses were achieved by ALI conditions. CONCLUSIONS: The ALICE-CLOUD system combined with lung epithelial cells cultured under ALI conditions offers a reliable and relevant in vitro method for preclinical aerosolized drug screening. Curcumin might be a promising anti-inflammatory candidate drug for inhalation therapy of lung diseases.

Simultaneous chiral analysis of amphetamine-type stimulants and ephedrine by capillary electrophoresis coupled to time-of-flight mass spectrometry.

This study focused on the chiral characteristics of methamphetamine seizures in Shanghai for inferring the synthetic pathways of drugs. Capillary electrophoresis coupled to time-of-flight mass spectrometry was used for simultaneous chiral separation of amphetamine-type stimulants and ephedrine, including S(+)-amphetamine/R(-)-amphetamine, S(+)-methamphetamine/R(-)-methamphetamine, (±)-MDA (3,4-methylenedioxyamphetamine), (±)-MDMA (3,4-methylenedioxymethamphetamine), (±)-MDEA (3,4-methylenedioxy-N-ethylamphetamine), d,l-N-ethylamphetamine, methylephedrine/methylpseudoephedrine, and 1S,2R(+)-ephedrine/(-)-ephedrine. The running buffer was 50-mM ammonium formate (pH 2.2 was adjusted by 1-M formic acid) containing 0.26% highly sulfated γ-cyclodextrin as the chiral selector. All enantiomers were well resolved within 40 minutes by capillary electrophoresis at 20 kV in an uncoated fused-silica capillary (50-µm I.D. × 375-µm O.D. × 90-cm length) and detected by micro time-of-flight mass spectrometry. Twenty seized methamphetamine samples were determined by the established method. They were classified into two groups through their chiral characteristics.

A simple and rapid infrared-assisted self enzymolysis extraction method for total flavonoid aglycones extraction from Scutellariae Radix and mechanism exploration.

A new, simple, and fast infrared-assisted self enzymolysis extraction (IRASEE) approach for the extraction of total flavonoid aglycones (TFA) mainly including baicalein, wogonin, and oroxylin A from Scutellariae Radix is presented to enhance extraction yield. Extraction enzymolysis temperature, enzymolysis liquid-to-solid ratio, enzymolysis pH, enzymolysis time and infrared power, the factors affecting IRASEE procedure, were investigated in a newly designed, temperature-controlled infrared-assisted extraction (TC-IRAE) system to acquire the optimum analysis conditions. The results illustrated that IRASEE possessed great advantages in terms of efficiency and time compared with other conventional extraction techniques. Furthermore, the mechanism of IRASEE was preliminarily explored by observing the microscopic change of the samples surface structures, studying the main chemical compositions change of the samples before and after extraction and investigating the kinetics and thermodynamics at three temperature levels during the IRASEE process. These findings revealed that IRASEE can destroy the surface microstructures to accelerate the mass transfer and reduce the activation energy to intensify the chemical process. This integrative study presents a simple, rapid, efficient, and environmental IRASEE method for TFA extraction which has promising prospects for other similar herbal medicines. Graphical Abstract ᅟ.

A comprehensive technology strategy for microbial identification and contamination investigation in the sterile drug manufacturing facility-a case study.

Objective: A comprehensive strategy for microbial identification and contamination investigation during sterile drug manufacturing was innovatively established in this study, mainly based on MALDI-TOF MS for the identification and complemented by sequencing technology on strain typing. Methods: It was implemented to monitor the bacterial contamination of a sterile drug manufacturing facility, including its bacterial distribution features and patterns. In three months, two hundred ninety-two samples were collected covering multiple critical components of raw materials, personnel, environment, and production water. Results: Based on our strategy, the bacterial profile across the production process was determined: 241/292 bacterial identities were obtained, and Staphylococcus spp. (40.25%), Micrococcus spp.(11.20%), Bacillus spp. (8.30%), Actinobacteria (5.81%), and Paenibacillus spp. (4.56%) are shown to be the most dominant microbial contaminants. With 75.8% species-level and 95.4% genus-level identification capability, MALDI-TOF MS was promising to be a first-line tool for environmental monitoring routine. Furthermore, to determine the source of the most frequently occurring Staphylococcus cohnii, which evidenced a widespread presence in the entire process, a more discriminating S. cohnii whole-genome SNP typing method was developed to track the transmission routes. Phylogenetic analysis based on SNP results indicated critical environment contamination is highly relevant to personnel flow in this case. The strain typing results provide robust and accurate information for the following risk assessment step and support effective preventive and corrective measures. Conclusion: In general, the strategy presented in this research will facilitate the development of improved production and environmental control processes for the pharmaceutical industry, and give insights about how to provide more sound and reliable evidence for the optimization of its control program.

[Study on the proficiency testing program of pharmaceutical preparation analysis].

This proficiency testing program is established to evaluate the pharmaceutical preparation analysis capacity of laboratories recommended by 18 countries and economies. It was authorized by Asia Pacific Laboratory Accreditation Cooperation (APLAC), and organized by Shanghai Institute for Food and Drug Control (SIFDC) and China National Accreditation Service for Conformity Assessment (CNAS). The 0.3sigma test is used to evaluate the homogeneity and stability of the proficiency testing sample. The results of the laboratories were assessed by Z-score. The robust average and the robust standard deviation of the participants' results were calculated as assigned value and standard deviation for performance assessment of hydrochlorothiazide and captopril using robust statistics. Thirty-three of 38 laboratories recommended by 18 countries and economies sent their results back. Twenty-four laboratories' results were observed as satisfactory. Five laboratories were identified as having reported at least one questionable result. Four laboratories were identified as having reported at least one unsatisfactory result.

[Local straight line screening method for the detection of Chinese proprietary medicines containing undeclared prescription drugs].

The present paper constructs a new approach named local straight-line screening (LSLS) to detect Chinese proprietary medicines (CPM) containing undeclared prescription drugs (UPD). Different from traditional methods used in analysis of multi-component spectrum, LSLS is proposed according to the characteristics of original infrared spectra of the UPD and suspected CPM, without any pattern recognition or concentration model establishment. Spectrum-subtraction leads to the variance in local straight line, which serves as a key in discrimination of whether suspected CPD is adulterated or not. Sibutramine hydrochloride, fenfluramine hydrochloride, sildenafil citrate and lovastatin were used as reference substances of UPD to analyze 16 suspected CPM samples. The results show that LSLS can obtain an accurate quantitative and qualitative analysis of suspected CPM. It is possible for the method to be potentially used in the preliminary screening of CPM containing possible UPD.

Hybridization-initiated exonuclease resistance strategy for simultaneous detection of multiple microRNAs.

Multiple miRNA detection is often limited by sample, time-consuming, and complicated procedures. To address such challenges, we present a relatively simple and amplification-free fluorescent strategy based on hybridization-initiated exonuclease resistance for simultaneous detection of multiple miRNAs in a single tube. Single-stranded linear DNA probes were designed with dual roles of capture and reporter DNA, i.e. each DNA probe was labeled with biotin at the 3'-end for signal readout and amino at the 5'-end for probe immobilization. After target miRNAs specifically hybridized with the corresponding probes, the formation of double-stranded probe-target duplex protected the biotin on the probe from Exonuclease I digestion, thus resulting in high fluorescent intensities through the reaction between biotin and streptavidin-phycoerythrin. Coupling with the 3-plex fluorescent microsphere-based assay system, herein we successfully demonstrated the simultaneous and quantitative measurement of three sequence-specific miRNAs at concentration range of 2.5 pM to 1.25 nM and detection limits of 2 pM. To meet high throughput and rapid turnaround time requirements, we have also exemplified that, even shortening the total reaction time within 1 h, wider linear response range and lower detection limit were guaranteed. We further applied this assay to detect endogenous target miRNA levels from five kinds of cancer cell line and one normal cell line HEK 293T. This simple and rapid strategy will hold great potential for monitoring of multiple miRNAs biomarkers in biomedical research and early clinical diagnosis.

[Discrimination of adulterated traditional Chinese medicines by infrared spectroscopy-two dimensional correlation analysis].

A new method was established, based on infrared spectroscopy two dimensional (2D) correlation analysis, for the discriminative analysis of adulteration in traditional Chinese medicines (TCM). Fenfluramine hydrochloride (FH) and sibutramine hydrochloride (SH) were taken as examples of synthetic drugs (adulterant), and the correlative peaks of their synchronous 2D correlation spectra were found. Then the characteristics of the synchronous 2D correlation spectrum of the suspected TCM were compared with those of FH. Since the correlative peaks in the synchronous spectrum of the suspected TCM coincide well with those of FH, a positive conclusion could be drawn after further investigation of the asynchronous spectra of TCM, which could provide the information about the source of correlative peaks. On the contrary, the dissimilarity of the synchronous spectra of SH and TCM directly implies that the suspected TCM is not adulterated with SH. The method can be used for a correct discrimination on whether the TCM is adulterated with the synthetic drugs, it does not rely on sample separation, and provides a new simple and cost-effective alternative to test the adulteration of TCM.

A simple and sensitive GC method for determination of N-methylpyrrolidine in cefepime and its preparation.

A new, simple and rapid gas chromatographic method was developed for the determination of N-methylpyrrolidine (NMP) in cefepime and its preparation. NMP was extracted with chloroform from cefepime and its preparation. An HP-1 column was maintained at 100 degrees C. Both the injector and the FI detector were set 250 degrees C. Pyridine was used as an internal standard. The detector response was linear up to 135 ng. The detection limit was 0.3 ng. The recoveries were 100.2-103.0% at three concentration levels. No interference from organic solvents presented in the synthesis was observed. The proposed method has a potential for application in quality control for cefepime and its preparation.

The LC-MS/MS methods for the determination of specific antibiotics residues in food matrices.

This chapter describes the LC-MS/MS methods for the determination of antibiotics residues in food matrices. The types of antibiotics include ß-lactam antibiotics, sulfonamides, tetracyclines, fluoroquinolones, nitrofurans, and chloramphenicol (CAP). The food matrices are mainly from animal origin, such as animal tissues, fishes (marine products), eggs, milk, honey, and so on. The methods and procedures are covered, including three parts: (1) Liquid chromatographic conditions, (2) mass spectrometer conditions, including ionization source, analyzer, and acquisition, and (3) extraction and clean-up methods. In each case, the standard operating procedures (SOPs) for analysis are given with sensitivity, linearity, precision, and recovery. Some criteria of maximum residue limits (MRLs) from the legislation are listed.

A new method for testing synthetic drugs adulterated in herbal medicines based on infrared spectroscopy.

Developing a simple and fast method to analyze possibly adulterated synthetic drugs in suspected herbal medicines (HM) is both methodologically and commercially significant. This paper constructs a new approach named local straight-line screening (LSLS), to the solution of the problem, after carefully observing the characteristics of the spectral line shapes. LSLS can be applied to both the qualitative and quantitative analysis of suspected HM, based solely on infrared spectrum of the possible synthetic drug and the suspected HM without any sample pretreatment. The concept and the application of the method are exemplified by analysis of sibutramine hydrochloride (SH), an anti-obesity medicine, adulterated in some commercially available HM-based diet pills, and compared with MSFDSS, and SNICA methods. The results show that, despite highly overlapping spectra and unresolved data, it is possible with LSLS to obtain an accurate discrimination and determination of SH in the HM-based diet pills. This allows the method to be potentially used in the primary screening of other synthetic drugs suspiciously adulterated in HM, with high rapidity, accuracy and cost-effectiveness.