Reproductive toxicity of JJH201501 in rats: Perinatal study.

INTRODUCTION: In this study, JJH201501 was examined for reproductive toxicity during the perinatal period to support its safety as a novel serotonergic agent (5-HT) antidepressant. Pregnant Sprague-Dawley rats (F0, n = 24/group) were continuously exposed to 0 (control), 6, 18, and 60 mg/kg body weight/day of JJH201501 by intragastric administration from gestation day 15 to lactation day 21. METHODS: During this period, maternal toxicity was evaluated based on clinical signs, body weight, feed intake, delivery condition, litter parameters, and necropsy, with body weight, sex ratios, malformation incidence, physical, and neurodevelopmental assessments conducted on all offspring rats. Ten pups (male:female 1:1) from each dam within each dose group on postnatal day 4 (PND4) were randomly selected. One pair was evaluated for behavior evaluations (F1a) after PND35, one for reproduction performance (F1b) after 10 weeks, and three for organ weight and deformities (F1c) on PND35. After successful mating, F1b male rats were weighed and dissected to assess reproductive organ weight and sperm motility. Pregnant F1b rats were weighed and monitored for food intake twice weekly until laparotomy on GD14, which recorded live/dead fetuses, resorptions, implantations, corpora lutea, and uterine weight. Some statistical differences were found between the JJH-treated and control groups in maternal weight, food consumption, and F1 body weight and water maze performance. RESULTS: Autopsy results showed that JJH201501 had a low cardiac index effect in F0, with no significant histopathological changes detected. Only one F1 offspring died in the high-dose group throughout the experiment. Due to the lack of dose-dependent effects and the consistent growth pattern of these alterations, the study findings do not suggest any toxicological significance for the observed results. CONCLUSION: In conclusion, the no-observed-adverse-effect level of JJH201501 for perinatal rats is about 60 mg/kg b.w./day.

A novel LC-MS/MS approach to [19 F]-alfatide and alfatide for application in preclinical pharmacokinetics.

For the further study of [19 F]-alfatide, the development of highly sensitive analytical methods for its determination is an urgent issue. In this paper, a method for simultaneously determining [19 F]-alfatide and alfatide using liquid chromatography/tandem mass spectrometry was created and validated. The plasma samples were pretreated using the protein precipitation method. Poroshell 120 EC-C18 (4.6 × 50 mm, 2.7 µm, Agilent) was used for the separation of the analytes to be measured. The mobile phase, consisting of acetonitrile and water (0.1% acetic acid, 50:50, v/v), was delivered at a flow rate of 0.60 ml/min for sample analysis. Positive electrospray ionization was performed using multiple-reaction monitoring with transitions of m/z 715.2 → 636.6 for [19 F]-alfatide, the ion pairs 700.3 → 851.5 for alfatide and the ion pairs 237.1 → 194.2 for carbamazepine (internal standard). According to the results, the method had high specificity, precision and accuracy as well as an extended linear range. The matrix effect and extraction recovery were also acceptable. The method was successfully applied to the pharmacokinetic study of [19 F]-alfatide in rats.

Embryo-fetal developmental toxicity and toxicokinetics of loxoprofen tromethamine intravenously administered to pregnant rats.

BACKGROUND: Loxoprofen tromethamine is a novel structural compound related to loxoprofen. It has been used for the treatment of pain and inflammation. However, the embryo-fetal developmental toxicity (EFDT) of loxoprofen tromethamine has not been evaluated in detail in vivo. This study investigated the EFDT and toxicokinetics of loxoprofen tromethamine in rats. METHODS: The aim of this study was to investigate the potential reproductive toxicity on embryo-fetal development of loxoprofen tromethamine (0, 1, 3, and 10 mg/kg/day) and sodium cyclophosphamide (CP) (2.8 mg/kg/day) administered by intravenous injection to pregnant rats during gestation days (GDs) 6-15. Pregnant rats were euthanized on GD20. The numbers of live/dead fetuses, resorptions, implantations, and corpora lutea, gravid uterus mass, placenta mass, fetal gender ratios, body weight, and skeletal development were evaluated. In a concomitant toxicokinetic (TK) study (10 pregnant rats per group), plasma TK parameters and the tissue distribution of loxoprofen tromethamine were tested. RESULTS: On GD20, rats were anesthetized and dissected by caesarean section. The appearance, internal organs, gravid uterus weight, embryo implantation number, and implantation loss rate in maternal rats of each group did not reveal any lesions. In fetuses, there were no significant differences in the fetus weight, embryo resorption number, stillbirth number, or fetal visceral examination in all test groups compared to the negative control group. However, in the high-dose group, the fetuses showed significant differences in the anomalies of the bones compared to the negative control group. The TK study showed that in the dose range of 1-10 mg/kg, the Cmax and AUC(0-t) of loxoprofen tromethamine in animals after the first administration increased proportionally to the dose, showing linear kinetic characteristics; after the last administration, the Cmax and AUC(0-t) increased disproportionately to the dose, showing nonlinear kinetic characteristics. The results of tissue distribution show that loxoprofen tromethamine was mainly distributed in the placenta and lung after the intravenous administration to pregnant rats; the content in the liver was lower and increased sharply in the heart with increasing doses; the content in all tissues was lower than that in the plasma. Loxoprofen tromethamine in fetal tissues and organs was mainly distributed in fetal lungs, liver and heart, and the lowest content was in amniotic fluid. CONCLUSIONS: In conclusion, the no-observed-adverse-effect level (NOAEL) and lowest-observed-adverse-effect level (LOAEL) of loxoprofen tromethamine were considered to be 1 and 10 mg/kg/day, respectively.

Simultaneous Determination of Moxifloxacin Hydrochloride and Dexamethasone Sodium Phosphate in Rabbit Ocular Tissues and Plasma by LC-MS/MS: Application for Pharmacokinetics Studies.

Treatment of ocular infection involves pharmacotherapy with steroids and antibiotic drops, such as moxifloxacin hydrochloride (MFH) and dexamethasone sodium phosphate (DSP). To characterize the pharmacokinetics of these two compounds, we performed and validated a liquid chromatography-mass spectrometry (LC-MS/MS) method to quantify them in rabbit ocular tissues and plasma. We used protein precipitation to extract the compounds. The analyte and internal standard (IS) were separated using a Shim-pack Scepter C18 column. The mobile phase was composed of 0.1% formic acid water (A) and methanol (B). MFH and DSP were detected using positive ion electrostatic ionization (ESI) in multiple reaction monitoring mode (MRM). The calibration curves for both compounds showed good linearity over concentrations ranging from 0.5 to 200 ng/mL in rabbit ocular tissues and plasma. The lower limit of quantification for both MFH and DSP was 0.5 ng/mL. We validated this method for selectivity, linearity (r2 > 0.99), precision, accuracy, matrix effects, and stability. Thus, we used this method to assess the pharmacokinetic (PK) characteristics of MFH and DSP in rabbit ocular tissues and plasma after single doses. Our results indicate that this method can be used for the simultaneous analysis of moxifloxacin hydrochloride and dexamethasone sodium phosphate in clinical samples.

Study on the intestinal permeability of lamivudine using Caco-2 cells monolayer and Single-pass intestinal perfusion.

Background: The aim of this work is to investigate the intestinal permeability of lamivudine and explore its absorption mechanism. Method: Caco-2 cells monolayer and single-pass intestinal perfusion (SPIP) were selected for the investigation of lamivudine under different conditions, such as different concentration, absorption time, bidirectional transportation, and transportation with efflux transporters inhibitor. The concentration of lamivudine both in Caco-2 cells monolayer samples and SPIP samples was detected by HPLC-UV. Then the permeability parameters were calculated. Results: The established HPLC-UV method reach the requirements for detection. There is no statistically difference between absorption parameters of lamivudine both in Caco-2 cells monolayer and SPIP (P > 0.05) under different dose groups. After transportation with efflux transporters inhibitor, the efflux rate of lamivudine in three dose groups was significantly decreased from 2.67, 2.59 and 2.59 to 1.78, 1.61, and 1.81 respectively. Lamivudine exhibits an absorption mechanism of passive diffusion. Conclusion: The absorption of lamivudine may be related to efflux transporters. In addition, lamivudine is a moderate-permeability drug in Biopharmaceutics Classification System.

Fertility and early embryonic development toxicity and toxicokinetic study of KFP-H008 in Sprague-Dawley rats.

The novel potassium competitive acid blocker, KFP-H008, developed to overcome the shortcomings of proton pump inhibitors. In this study, KFP-H008 was administered by oral gavage at doses of 0 (control), 15, 45, and 150 mg/kg to Sprague-Dawley rats (24 animals per sex per group). Body weight, food consumption, mortality, sexual cycle, mating behavior, pregnancy, sperm motility, and relative organ weights were evaluated. In a concomitant toxicokinetic (TK) study (10 animals per sex per group), plasma TK parameters and tissue distribution of KFP-H008 were tested. There were obvious effects at the dosage of 150 mg/kg to male rats with decreases of prostate weight and lower weight gain; to female rats with lower weight gain, hair off, and lower corpus luteum count. There were no effects on litter parameters. Time to reach maximum plasma concentrations for KFP-H008 was between 0.5 and 1.0 hr for the 15 and 45 mg/kg groups, while for the 150 mg/kg group it had a delay to 2 hr. Overall, the NOAEL of KFP-H008 was considered to be 45 mg/kg in both genders and the TK study shows that exposure (area under the curve) of male rats was about 1,091.0 ± 530.8 µg/Lhr and to female rats was 1,030.5 ± 512.5 µg/Lhr. Administration of KFP-H008although reaches the reproductive organs, it demonstrated no significant effects on reproductive organs, it did not affect mating, fertility, pregnancy, growth, or morphologic development.

Flow synthesis, characterization, anticoagulant activity of xylan sulfate from sugarcane bagasse.

High-value utilization of hemicellulose is critical to improve the append value of integrated biorefineries. In this research, the alkali-soluble sugarcane bagasse hemicellulose was sulfated using chlorosulfonic acid and N,N-dimethylformamide/LiCl under homogeneous conditions. With the aid of flow technique, a rapid, mild, and efficient method for the synthesis of xylan sulfate with high molecular weight and controllable degree of substitution was achieved. The results showed that the reaction time and the degradation of xylan chain were drastically reduced compared to the "in flask" batch conditions. High molecular weight of the product (Mw = 148,217) with a reasonable degree of substitution (DS = 1.49) could be obtained even at room temperature in 10 min under the present flow system. Anticoagulant experiments showed good anticoagulant activity of the resultant xylan sulfate, which could significantly prolong the activated partial thromboplastin time and thrombin time. This work not only provides a novel method for the synthesis of xylan sulfate, but also offers new opportunities for the production of other functional polysaccharide derivatives under the flow reaction conditions.

An embryo-fetal development toxicity study with dimethylaminoethyl ginkgolide B in rats and rabbits.

Ginkgo biloba (a herbal product) has long been used in traditional Chinese medicine to treat cerebrovascular and cardiovascular diseases. Ginkgolide B is one of the important pharmacologically active components of Ginkgo biloba. Dimethylaminoethyl ginkgolide B (a novel ginkgolide B derivative) has been developed to overcome the poor water-solubility of natural drugs and to achieve higher bioavailability. This study investigated the potential effects of dimethylaminoethyl ginkgolide B on pregnant dams and embryo-fetal development in Sprague-Dawley rats and New Zealand white rabbits following maternal exposure on gestational day (GD) 6-15 and GD 6-18, respectively. Dimethylaminoethyl ginkgolide B was administered by intravenous injection to pregnant rats (0, 10, 30 and 100 mg kg-1 d-1) and rabbits (0, 6, 18 and 60 mg kg-1 d-1). Maternal toxicity signs, such as lower maternal body weight gain and food consumption, were observed at 100 mg kg-1 d-1 in rats and 60 mg kg-1 d-1 in rabbits. The developmental toxic effects included a decrease in fetal and placental weights, increased incidences of skeletal variations and delay in fetal ossification. Fetal growth and development were affected by dimethylaminoethyl ginkgolide B in the high-dose group in rabbits. The no-observed-adverse-effect level of dimethylaminoethyl ginkgolide B is considered to be 30 mg kg-1 d-1 for rats and 18 mg kg-1 d-1 for rabbits.

Embryo-fetal toxicity assessment of vonoprazan in rats and rabbits.

Vonoprazan is a new potassium-competitive acid blocker to treat acid-related diseases. However, its safety during pregnancy is unclear. The aim of the study was to investigate the potential reproductive toxicity on the embryo-fetal development of vonoprazan. Vonoprazan acetate was administered by intravenous injection to pregnant rats (0, 2, 6 and 20 mg kg-1 day-1 ) and rabbits (0, 1.2, 3.6 and 12 mg kg-1 day-1 ) during the organogenetic period (gestation day 6-15 [rats] and 6-18 [rabbits]). Maternal reproductive endpoints were evaluated, together with effects on fetal growth and morphological development. In rats, no treatment-related effects were found in the highest dose group (20 mg kg-1 ) and the maternal plasma exposure was ≥50-fold the expected clinical human exposure. However, in rabbits, dose-related clinical signs (soft or liquid feces) occurred in the 12 mg kg-1 group, which was regarded as a maternal toxicity. Besides, decreased maternal weight gain also was considered as a minimal maternal toxicity. At 12 mg kg-1 , delayed fetal ossification was found as evidence of embryo-fetal growth retardation, which was related to decreased fetal and placental weights. There was no maternal and developmental toxicity in the 1.2 and 3.6 mg kg-1 groups. Thus, the no-observed-adverse-effect levels of vonoprazan acetate in rabbits are considered 3.6 mg kg-1 day-1 , which produced plasma exposure that was about 18-fold human clinical exposure.

[Immunogenicity of porcine fibrinogen].

OBJECTIVE: To investigate the immunogenicity and immune effect of porcine fibrinogen and provide experimental evidence for safety evaluation and further optimization of the drug. METHODS: An improved ABC-ELISA system of biotin and conjugated streptomyces avidin was used for detecting antibodies. The serum antibody titer and its dynamics were studied by means of the injury experiment in New Zealand rabbits. Effects of porcine fibrinogen on immune system of mice were evaluated by analyzing the weight gain and indexes of immune organs, the proliferation of spleen lymphocytes by thiazole blue (MTT) chromatometry and the neutrophil phagocytosis by nitro-tetrazolium chloride blue (NTB) reduction method. RESULTS: The antibodies could be detected in New Zealand white rabbit serum 1 week after immunization, and antibody levels decreased to a more stable level after it peaked at the second week, at the same time, the antibody titer also showed the same trend. The weight and indexes of immune organs were not changed significantly in mice. Compared with the control group, the proliferation rate of lymphocytes in each experimental group didn't show statistical difference, but neutrophil phagocytosis of the experimental groups was enhanced significantly. CONCLUSION: The humoral immune response was induced in New Zealand rabbits after stimulated with porcine fibrinogen, but no specific immune response was found in mice.

Past, present, and future industrial biotechnology in China.

Fossil resources, i.e. concentrated carbon from biomass, have been irrecoverably exhausted through modern industrial civilization in the last two hundred years. Serious consequences including crises in resources, environment and energy, as well as the pressing need for direct and indirect exploitation of solar energy, pose challenges to the science and technology community of today. Bioenergy, bulk chemicals, and biomaterials could be produced from renewable biomass in a biorefinery via biocatalysis. These sustainable industries will match the global mass cycle, creating a new form of civilization with new industries and agriculture driven by solar energy. Industrial biotechnology is the dynamo of a bioeconomy, leading to a new protocol for production of energy, bulk chemicals, and materials. This new mode of innovation will place the industry at center stage supported by universities and research institutes. Creativity in industrial biotechnology will be promoted and China will successfully follow the road to green modernization. China's rapid economic development and its traditional capacity in fermentation will place it in an advantageous position in the industrial biotechnology revolution. The development and current status of industrial biotechnology in China are summarized herein.

Simultaneous Determination of Four Active Components in Tobacco Wastes by LC.

A liquid chromatographic method was developed for the simultaneous quantification of four major active components in tobacco (Nicotiana tobaccum L.) wastes. Samples were extracted with 70% v/v aqueous methanol, four compounds including chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid and caffeic acid were identified and determined by using LC coupled to electrospray tandem mass spectrometry and LC-UV method, respectively. Separation in LC-UV was on an Alltima C18 column (250 mm × 4.6 mm i.d.; 5 µm) with a mobile phase consisting acetonitrile: ammonium acetate buffer (pH 4.5) (5:95 v/v), at a flow rate of 1.0 mL min-1, detected at 327 nm. Four regression equations showed good linear relationships (r 2 > 0.999) between the peak area of each marker and concentration. The method has good repeatability and precision, the intra-day and inter-day RSD for both retention time and peak area was less than 1.0%. The recoveries, measured at three concentration levels, varied from 96.33 to 101.10%. The LOD (S/N = 3) and LOQ (S/N = 6) were less than 0.010 and 0.795 µg·mL-1, respectively. This assay was successfully applied to the determination of four active compounds in ten samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quantitative analysis method for various of tobacco wastes.