In recent years therapeutic proteins products including therapeutic antibodies have become a major driving force for the modern biopharmaceutical industry. However, they have complex product quality attributes (PQAs) which limit product development and quality control (QC). Recent advances in high resolution mass spectrometry (MS) have led to the use of an MS-based multi-attribute method (MAM) for quality control testing of therapeutic proteins, which allows for direct measurement of multiple PQAs and identification of impurities. MAM helps to promote the improvement of product quality and QC and a reduction in manufacturing cost. To explore the application of MAM in QC, we discuss generic MAM workflow, the current state of MAM application in product development and QC, identify points to consider for use of MAM as a QC test, and summarize MAM's advantages and challenges in this article. The future application of MAM for therapeutic antibodies and the opportunities for its further development, use, and substitution for conventional methods is presented.

The treatment plan for chronic pain often proceeds from a single drug to drug combination therapy. Sinomenine and ligustrazine, natural alkaline substances derived from traditional Chinese medicines, are expected to provide a new choice for combination analgesic therapy strategies. Here we establish a microdialysis sampling and HPLC-MS/MS quantification method for sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rat blood and brain extracellular fluid. Blood and brain microdialysis probes were implanted in the jugular vein toward the right atrium and left corpus striatum zone (AP +0.2 mm, ML 3.0 mm, DV 3.5 mm) in rats. The blood and brain microdialysis probes were perfused with citric acid buffer solution and Ringer's solution, respectively. Blood and brain extracellular fluid microdialysate were collected at intervals of 20 min at a perfusion rate of 1.5 μL·min^-1, and continuously collected for 24 h after administration. The liquid chromatographic separation used a C18-reversed phase chromatographic column (HSS T3 2.5 μm, 2.1 mm×50 mm), the mobile phase was methanol/water (containing 0.05‰ formic acid), and gradient elution was carried out at a flow rate of 0.3 mL·min^-1. Mass spectrometric detection used an electrospray ion source, positive ion mode and multi-reaction monitoring method. The selected quantitative ions for sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin, amitriptyline and internal standard naloxone were 330/181, 137/80, 172/154, 152/110, 160/142, 278/233 and 328/310 respectively. The specificity, linear range, matrix effect, accuracy, precision, stability and probe recovery were investigated and confirmed to be suitable for the determination of the above drugs in rat blood and brain extracellular fluid microdialysate. The calculated in vivo recovery of microdialysis probes ranged from 19.38% to 25.88%. After intravenous administration of sinomenine (50 mg·kg^-1), ligustrazine (50 mg·kg^-1), gabapentin (50 mg·kg^-1), paracetamol (50 mg·kg^-1), pregabalin (50 mg·kg^-1) and amitriptyline (40 mg·kg^-1) to rats, the peak concentration in the blood microdialysate was in the range of 0.2-10 μg·mL^-1. Drug concentrations could also be detected in brain extracellular fluid microdialysate, however with lower levels (peak concentration:0.1-6 μg·mL^-1) than those of blood microdialysates at each time point. In conclusion, this method can be applied to microdialysis sampling and quantification of sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rats. The method will promote research in identifying herb-drug pharmacokinetic interactions, as well as safety concerns in combination-therapy strategies.

This research explored the synergistic effects and the mechanism of parthenolide (PTL) and vorinostat (suberoylanilide hydroxamic acid, SAHA) on the proliferation of A549 non-small cell lung cancer cells. The combination effect of PTL and SAHA was detected by cell counting kit-8 (CCK-8) and colony formation assays. Scratch test was performed to detect cell migration. Annexin V-fluorescein isothiocyanate isomer/propidium iodide (FITC/PI) flow cytometry and Western blot analyses were used to determine cell apoptosis and its mechanism. The results showed that combination of PTL and SAHA inhibited the proliferation and migration of A549 with a synergistic effect compared to the single-drug groups. The combination of PTL and SAHA had synergistic effect to induce cell apoptosis by regulating p53 and c-myc pathways, and affected the expression levels of poly ADP-ribose polymerase (PARP), cysteinyl aspartate specific proteinase (caspase)-9, and caspase-3. Taken together, this study shows that combination of PTL and SAHA has synergistic effect, induces cell apoptosis and inhibits A549 proliferation, which is likely to be a novel strategy for the treatment of non-small cell lung cancer.

Cisplatin is one of the most commonly used chemotherapeutic drugs in clinic and has good therapeutic effect on various cancers, but the development of drug resistance limits its clinical treatment. The development of cisplatin resistance is caused by many factors, including the decrease of intracellular cisplatin accumulation, the inactivation of cisplatin by mercaptan proteins, the increase of DNA damage repair, apoptosis inhibition, tumor microenvironment and cancer stem cells. In recent years, traditional Chinese medicine (TCM) has been favored for its remarkable effect of reversing cisplatin resistance. This review will explore the mechanisms of cisplatin resistance and the combined modality treatment strategy of TCM to reverse cisplatin resistance, hoping to provide reference for clinical and scientific research.

Tanshinones and salvianolic acids are important materials in the treatment of coronary heart disease, myocardial infarction, hypertension, hyperlipidemia, stroke and others illnesses. In recent years, with the development of genomics, transcriptome, metabolomics and bioinformatics, many advances have been made in the biosynthesis and transcriptional regulation of tanshinones and salvianolic acids. This paper summarizes these advances and suggests further study on the downstream synthesis pathways and transcriptional regulatory mechanisms to reveal new molecular mechanism of synthesis, transport, regulation and modification. Additionally, we discuss the design and construction of new biological pathways to increase the expression of biosynthesis genes and the production of secondary components, is a newly developing research field.

New candidate targets, biological mechanisms as well as small-molecules are significant factors in the research and development of first-in-class drugs, which is a challenging process with a large amount of time and money devoted as well as high risks. A successful first-in-class drug can not only become a new strategy for disease treatment but can also offer innovative research ideas for the design of drugs. The Food and Drug Administration (FDA) approved 48 new drugs to the market in 2019, among which small-molecule drugs still predominated, containing several first-in-class drugs. Brexanolone, for example, is the first positive modulator of GABA_A receptor for the treatment of postpartum depression; Selinexor is the first small-molecule drug to treat recurrent refractory multiple myeloma by inhibiting exportin (XPO1); Tenapanor is the first sodium/proton exchanger type 3 (NHE3) inhibitor that can treat irritable bowel syndrome; Lasmiditan is the first approved agonist with selectivity for 5-HT_1F, treating migraines. The research and development processes of the first-in-class drugs mentioned above are distinctive from each other with uniqueness and innovation. In this review, we briefly analyze the background and process of the research and development of three typical cases as well as their therapeutic applications in an attempt to offer some help for the future development of first-in-class drugs.

In order to achieve rapid proliferation and adapt to the complex microenvironment, tumor cells have dominant characteristics such as unique metabolic patterns and the ability to escape from immunoregulation. Tumor cells reprogram multiple metabolic pathways to promote immune escape, which impacts tumor diagnosis, treatment and prognosis. Based on the effect of metabolic changes on tumor immune escape and its molecular mechanism, metabolic regulation provides new approaches to enhance immunotherapy. We review recent advances in tumor immuno-escape and immunotherapy based on metabolic regulation. Cutting-edge analytical techniques and methods for tumor metabolism research such as metabolomics, mass spectrometry imaging-based spatially-resolved metabolomics and metabolic flow analysis are also discussed.

Metabonomics techniques were used to investigate the mechanism and metabolic pathways of total extract of Amygdalus mongolicus against renal fibrosis in rats. Rats were randomly divided into a model group (MOD), a sham surgery group (SDG), a benazepril hydrochloride-treated group (BHT) and three groups treated with the total extract of Amygdalus mongolicus:low-dose group (TOT-L), middle-dose group (TOT-M) and high-dose group (TOT-H), with 10 rats in each group. The rats were given intragastric administration for 3 weeks and kidney and blood samples were taken. Pharmacodynamic studies and ultra performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-Q-TOF/MS) analysis were used to show that the total extract of Amygdalus mongolicus has an anti-fibrotic effect in rats. Compared with the MOD group, rats in the TOT-L, TOT-M and TOT-H groups showed a reversal in 67, 69, and 70 biomarkers, respectively, and shared 62 biomarkers. Reversal was observed for 7 key biomarkers related to renal fibrosis, including S-adenosy-L-methioninamine, ornithine, diketogulonic acid, and others, and changes in 5 metabolic pathways, including arginine and proline metabolism, pentose and glucuronate interconversions. These results give evidence of the metabolic pathways and the mechanism of action of Amygdalus mongolicus to prevent renal fibrosis in rats. The animal experiments were approved by the Medical Ethics Committee of Baotou Medical College (No. 20190314).

In scientific research, it is often needed to knock in, knock out, knock down, or overexpress a specific gene in model organisms or specific types of cells to achieve precise regulation of experimental independent variables. In this case, various transgenic mice are required. The cyclization recombinase (Cre) can directly interact with different loxP (locus X over P1) DNA sequences without any cofactors to perform specific gene knock-in or knock-out at specific targets. Because of its advantages of simple action principles, high spatial specificity, and high reorganization efficiency, the Cre-loxP system is widely used in scientific research. Furthermore, the CreERT2 system (mutant of the fusion protein of Cre and estrogen receptor ligand binding domain) and the tetracycline (Tet)-on/off system, derived from the Cre-loxP system, have made the recombination of the target gene occur in temporal-specificity on the basis of spatial-specificity. This dual specificity of time and space is indispensable for research in specific directions such as fear memory and engram cells on the basis of reducing the impacts on experimental animals. Therefore, these derived systems have broad application prospects.

This study was designed to investigate the effect of dihydromyricetin (DHM) on inducing apoptosis of ovarian cancer cells A2780 through endoplasmic reticulum stress (ERS) pathway and the mechanisms involved in vitro and in vivo. A2780 cells were treated with different concentrations of DHM, and the protein expression levels of glucose-regulated protein 78 (GRP78) which is related to ERS increased, apoptotic proteins C/EBP-homologous protein (CHOP), and cysteinyl aspartate specific proteinase-12 (caspase-12) elevated. After pretreatment with ERS inhibitor, 4-phenyl butyric acid (4-PBA), following the intervention with DHM, the A2780 cell viability decreased and apoptotic rate increased. All animal welfare and experimental procedures were approved by the Animal Ethics Committee of Chongqing Medical University. Intraperitoneal injection of DHM suspension into nude mice with ovarian cancer could significantly inhibit the growth of transplanted tumor in vivo, increase the protein expression levels of GRP78, CHOP, and caspase-3. Moreover, swollen and broken endoplasmic reticulum could be observed in tumor tissues, suggesting that DHM intervention induces apoptosis mediated by ERS. The results indicated that DHM could induce apoptosis of ovarian cancer cells and inhibit the growth of transplanted tumors in nude mice, which might be related to the activation of ERS pathway.