An LC-MS/MS method was developed for the simultaneous determination of fosaprepitant and aprepitant in human plasma, and applied to a pharmacokinetic study of 150 mg fosaprepitant dimeglumine injection to 12 Chinese healthy volunteers. The analytes and internal standards were extracted from plasma by protein precipitation with acetonitrile and separated on a Cortex C18+ (50 mm×2.1 mm, 2.7 μm) column using a gradient elution procedure. Mass spectrometry was performed in negative MRM mode, and parent-to-produce transitions were as follows:m/z 613.1→78.9 for fosaprepitant, m/z 617.0→78.9 for d₄-fosaprepitant, m/z 533.2→275.1 for aprepitant and m/z 537.2→279.1 for d₄-aprepitant. Plasma sample was basified to stabilize fosaprepitant. The standard curves were demonstrated to be liner in the range of 15.0 to 6 000 ng·mL^-1 for fosaprepitant and 10.0 to 4 000 ng·mL^-1 for aprepitant. The intra-day precisions and inter-day precisions and accuracy were within the acceptable limits for all concentrations.

The Chinese herbal Sophora alopecuroides is widely used to clean intestine and eliminate dampness, and it has good therapeutic effects on treating bacillary dysentery and inflammatory bowel disease, etc. in clinics. However, the mechanism of treatment is not yet well understood. The present study was aimed to explore the mechanism of Sophora alopecuroides treatment of large intestine dampness-heat syndrome (LIDHS). The LIDHS model was performed by the comprehensive factors, including high temperature and humidity environment, high-sugar and high-fat diet, and intraperitoneal injection of Escherichia coli. The blood routine, serum proinflammatory cytokine levels and histopathological changes of intestine were detected and observed. Meanwhile, the serum metabolomic approach was conducted using the method of ultra performance liquid chromatography coupled to quadrupole time-of-flight mass/mass spectrometry (UHPLC-Q/TOF-MS/MS). The results showed that Sophora alopecuroides has good therapeutic effects on the LIDHS rat models. After treatment with Sophora alopecuroides, the abnormality of blood routine indexes as well as proinflammatory cytokines, including IL-1β, IL-2, IL-6 and TNF-α in vivo, tended to be normal, and the histopathological changes of intestine were improved. Through metabolic profiling and protocol analysis, 9 potential metabolic markers may be closely related with the treatment mechanism of Sophora alopecuroides on this disease, including taurine, L-tryptophan, LysoPE, LysoPC, LPA, DG, chenodeoxycholic acid disulfate, traumatic acid and 7-ketodeoxycholic acid, which were involved in taurine and hypotaurine metabolism, glycerophospholipid metabolism, glycerolipid metabolism, tryptophan metabolism and primary bile acid biosynthesis etc. The serum metabolomic approach can be applied to clarify the therapeutic mechanism of Sophora alopecuroides on LIDHS, and provide the theoretical basis for Sophora alopecuroides in clinical practice.

This study was designed to explore the mechanism of Coix seed oil (Coix) impact on the drug resistance, bioluminescence imaging (BLI) and the efflux of D-luciferin potassium salt, the substrate of ABC transporters, in doxorubicin-resistant breast cancer cells. Multidrug resistance (MDR) gene and protein expression were analyzed in the cells by q-PCR and Western blot. First, in order to investigate the effect of the efflux function by ABC protein, a cell line with overexpressed luciferase was established in MCF-7 cell line. BLI was used to monitor the efflux kinetics of D-luciferin potassium salt before and after Coix treament. The results showed that the efflux of D-fluorescein potassium from MCF-7/DOX^Fluc was lessened when pretreated with Coix, which means that Coix may decrease the efflux of other chemotherapies using ABC transporters. Both of the results of q-PCR and Western blot showed that gene and protein expression of ABC transporters such as ABCG2, ABCC1 and ABCB1 were down-regulated by Coix treatment. The efficacy of Coix reversing MDR was verified with the chemotherapy medication doxorubicin (DOX). MTT assay showed that Coix increased the inhibitory effect of DOX on proliferation of MCF-7/DOX, and the optimal combination of ratio was 25 times that of DOX. The results suggest that Coix may reverse MDR of the substrate of ABC transporters from two aspects, one is to cut down the ABC protein efflux function, and the other is to decrease the quantity of ABC gene and protein expression.

Compared with the racemate of chiral drugs, enantiopure chiral drugs have been the hot spot of drug research because of their higher selectivity and lower side-effects. Although remarkable progress of asymmetric synthesis has been achieved in the last decades, chiral resolution is regarded as an important approach to obtain chiral drugs. Recent research advancements in the field of chiral resolution of racemic drugs and intermediates are reviewed here. It is clear that combination of chiral separation and racemization to improve the resolution efficiency has become a trend of chiral resolution. In addition, we also introduce some novel resolution methods, such as chiral extraction, membrane resolution, and resolution using nanoparticles.

CRISPR/Cas9 system, consisting of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins, is a prokaryotic immune system that confers resistance to foreign genetic elements such as those present within plasmids and phages. A simple version of the CRISPR/Cas system, type Ⅱ CRISPR, has been modified to edit genomes. By delivering the Cas9 nuclease together with a synthetic guide RNA (sgRNA) into cells, genome can be edited at desired loci site. CRISPR/Cas genome editing techniques have been widely implemented in various species and research areas. In this review, we summarize the several applications of CRISPR/Cas9 in the field of drug discovery and development, which include target gene screening and editing, drug target screening and validation, generation of animal models and treatment of genetic disease, etc. The defects and improvements of CRISPR/Cas9 technology is discussed as well.

Currently, the specification grading standard for Astragali Radix can not accurately reflect growth years. The aim of this study is to identify the growth ring number of different parts of 1 to 6 year Hengshan imitative wild culture Astragali Radix, in order to get a different absolute growth years, to classify the accumulation rules of the content of flavonoids and saponins, and to lay the foundation for evaluating quality of Astragali Radix. Observing growth ring numbers of 1-6 years Astragali Radix by means of hand sections and paraffin sections in the study, and analyzing the number of different growth years and different diameter. At the same time, HPLC-UV-ELSD was used to analyze the 12 index components of the samples with absolute growth years of 2 to 6 years. The results indicated that the growth ring number excepting hollow part is consistent with the actual growth period of Astragali Radix and the number of growth rings gradually decreased from the upper to lower. The results of HPLC-UV-ELSD determination showed that the saponins content of 3-year-old Astragali Radix was the highest while the flavonoids content of the 4-year-old reached the maximum. The study provided the basis for foundation of the specification grading standard for Astragali Radix and clinical rational use drug.

Safflower is a dried flower of the annual herbaceous plant safflower (Carthamus tinctorius L.). As a traditional Chinese medicine, it was widely used in the regulation of blood circulation. Flavonoids are the main active ingredients in safflower. MYB transcription factors are involved in the regulation of flavonoids. The cloning and expression analysis of MYB transcription factor genes in safflower is of great significance, not only for clarifying the regulation mechanism of flavonoids biosynthesis in safflower, but also for the artificial regulation of flavonoid biosynthesis in safflower. Based on the transcriptome data, we used iTAK to annotate the MYB transcription factors in safflower. The MYB transcription factors were cloned and their sequences were analyzed. Besides, their expressions were analyzed by a Real-time PCR. In the experiment, eight long fragment MYB transcription factors were screened and six MYB transcription factors was successfully cloned, named CtMYB-TF1, CtMYB-TF2, CtMYB-TF4, CtMYB-TF5, CtMYB-TF6 and CtMYB-TF7, respectively. The six MYB transcription factors had the core domain of MYB transcription factor family, and evolutionary analysis showed that the CtMYB-TF7 transcription factor was closely related to the factors AtMYBL2 and AtMYB12. Expression analysis showed that the expression of CtMYB-TF5, CtMYB-TF6 and CtMYB-TF7 was low in roots, stems and leaves, and was high in the flower. The results provide a foundation for study of mechanism of molecular regulation of safflower flavonoids.

Honokiol (HK) have extensive pharmacological activities, but its poor solubility and instability restricted its clinical application and efficacy exertion. HK nanosuspensions (HK-NSps) were designed in this study in order to solve the problems. HK-NSps were prepared by antisolvent precipitation method, using poly-vinylpyrrolidone (PVP) and bovine serum albumin (BSA) as a combined stabilizer. The particle size was measured using dynamic light scattering method, the morphology was observed by transmission electron microscopy. The size change and drug content of HK-NSps in various physiological media during the storage at ambient temperature was examined to evaluate their storage stability. Dialysis method was used to study their drug release in vitro. MTT assay was used to assess their in vitro cytotoxicity against 4T1 breast cancer cell line. Anti-tumor effect in vivo was also investigated in 4T1 tumor-bearing mice. HK-NSps were prepared with high drug loading content of 48.62%, nearly spherical shape and good storage stability. The average particle size was (83.40 ±1.042) nm, the polydispersity index (PDI) value was 0.223 ±0.011, the zeta potential was (-42.2 ±1.2) mV. HK-NSps showed sustained in vitro drug release and enhanced cytotoxicity in contrast to free HK against 4T1 cells (IC₅₀, 8.36 μg·mL^-1 vs 37.58 μg·mL^-1, P < 0.05). The in vivo study on 4T1 tumor-bearing mice demonstrated that HK-NSps showed good dose-dependent tumor inhibition rate (TIR). In contrast to 4 mg·kg^-1 of PTX injection (TIR, 47.9%), medium and high dose of HK-NSps displayed improved therapeutic efficacy (TIR, 55.67% for 40 mg·kg^-1, 67.28% for 60 mg·kg^-1, P < 0.05). In contrast, the high dose of HK crude drug (60 mg·kg^-1) had TIR of only 54.13% even administrated every day. In conclusion, HK-NSps were prepared with small size, high drug-loading capacity, and good stability. The improved in vitro and in vivo antitumor efficacy demonstrated that HK can be a promising antitumor drug in combination with nanosuspensions technology.

Ginsenoside Rg1 is one of the main active components of ginseng with various pharmacological activities including anti-inflammatory, anti-oxidation, anti-aging, anti-tumor and anti-apoptosis. Ginsenoside Rg1 plays a protective role in multiple tissues and organs, which shows the multiple targeting properties of the pharmacological effects. Recently, a number of studies have demonstrated that ginsenoside Rg1 has a protective role in the liver due to its multiple pharmacological effects. In chemical liver injury models, or in other liver injury models, ginsenoside Rg1 can alleviate liver necrosis induced by oxidative stress and inflam-mation. This article provides a review of the recent studies on the efficacy of ginsenoside Rg1 in the treatment of various liver damage and the molecular mechanism.

This study was carried out to investigate the pharmacokinetics/bioequivalence of levornidazole disodium phosphate by using stable isotope labeled drug, evaluated the pharmacokinetic profile and confirmed the prodrug characteristics of levornidazole disodium phosphate in monkey. Levornidazole (Drug A) and stable isotope ¹⁵N labeled levornidazole disodium phosphate (Drug B) were mixed with equal mole amount (experiment Ⅰ); stable isotope ¹⁵N labeled levornidazole disodium phosphate (Drug B) and levornidazole disodium phosphate (Drug C) were mixed with equal mole amount, respectively. After giving the mixed drugs to the monkey, the concentration of ¹⁵N-levornidazole disodium phosphate, levornidazole disodium phosphate, ¹⁵N-levornidazole and levornidazole in plasma samples of pre-dosing and 24 h after administration were analyzed by a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method. Pharmacokinetic calculations were performed through non-compartmental analysis using WinNonlin software. Two-sided 90% confidence intervals (CI) were used to evaluate the bioequivalence of two drugs. The results showed that levornidazole disodium phosphate was metabolized to levornidazole rapidly after administration, the body exposure were increased with the dosage. The method of bioequivalence used in this study was different from the traditional two periods, crossover design. By using the method of this study, the effects of administration period, intra-individual variability, and sequence of administration on bioequivalence were avoided. The results of this study had successfully supported the pharmacokinetic and bioequivalence study of this drug in human using the same approach.