Guo Cheng

In vitro permeability analysis, pharmacokinetic and brain distribution study in mice of imperatorin, isoimperatorin and cnidilin in Radix Angelicae Dahuricae.

Coumarins are important constituents of Radix Angelicae Dahuricae, a well-known traditional Chinese medicine possess several known bioactivities with potentials in the treatment of central nervous system diseases. By using an HPLC-MS/MS method, we analyzed the in vivo plasma and brain pharmacokinetics of three ingredients of coumarins, including imperatorin, isoimperatorin and cnidilin in mice after oral administration of Dahuricae extract at doses of 800mg/kg. The biosamples were prepared using acetonitrile precipitation and the separation was achieved on an XDB-C18 column by gradient elution. The BBB permeability and P-gp-mediated efflux were further examined in Madin Canine kidney cells transfected with full length cDNA for human multidrug resistance gene1 (MDCKII-MDR1). Our results demonstrate that the method has excellent and satisfactory selectivity, sensitivity, linearity, precision, and accuracy for simultaneous determination of imperatorin, isoimperatorin and cnidilin. The pharmacokinetics parameters were determined by using noncompartmental analyses, including the AUC(0-t) in plasma (1695.22, 1326.45 and 636.98mg*h/L), the AUC(0-t) in brain (1812.35, 2125.17 and 1145.83ng*h/g) as well as the T1/2 in plasma (0.66, 0.82, 0.97h) and brain (0.96, 1.1, 0.99h) for imperatorin, isoimperatorin and cnidilin, respectively, suggesting that the three coumarins could easily pass through the BBB in vivo. In the in vitro model we observed high permeability of imperatorin and isoimperatorin with the P-gp-mediated efflux ratios of 0.53 and 0.06, as well as medium permeability of cnidilin with 0.82. All data suggest that these three coumarins have high BBB permeability and have pharmacokinetic potentials for the treatment of central nervous system diseases.

Parthenolide from Parthenium integrifolium reduces tumor burden and alleviate cachexia symptoms in the murine CT-26 model of colorectal carcinoma

Excessive elaboration of proinflammatory cytokines are involved in cachexia-related hypercatabolism. Parthenolide, as a potential anti-inflammatory active agent, could effectively inhibit nuclear factor-kappa B, and has the potential for the treatment of cancer cachexia. In this study, the cancer cachexia model was established by subcutaneous transplantation CT26 tumor fragment. Parthenolide or placebo was intraperitoneally given daily from the next day. Parthenolide treatment could effectively preserve the body weight, improve the mass of gastrocnemius and tibialis anterior muscles, and alleviate tumor burden. Sizes of muscle fibers and myosin heavy chain were also increasing. The serum proinflammatory cytokine TNF-α level was lower than placebo treatment mice measure by ELISA. To investigate the possible mechanism, MuRF1 and Fbx32 was subjected to Western blot analysis and expression of MURF1 was inhibited in gastrocnemius muscle. Collectively, parthenolide treatment could effectively alleviate tumor burden of cachexia, preserve the body weight and improve skeletal muscle characteristics.

NMR-based metabolomics reveals distinct pathways mediated by curcumin in cachexia mice bearing CT26 tumor

Background: cachexia is common in cancer patients, with profound metabolic abnormalities in response to malignant growth of cancer and progressive catabolism of host. Previous studies showed pharmacodynamics efficacy of curcumin in the prevention and treatment of cancer cachexia. However, the metabolic regulation effect is still unknown. Methods: we employed a proton NMR-metabonomics method to investigate the metabolic features of cancer cachexia and the contribution of curcumin to serum metabolites in a mouse model bearing CT26 tumor. Results: curcumin treatment (200 mg per kg per day) resulted in 13.9% less body weight loss and conserved mass of epididymal fat, muscle gastrocnemius and muscle tibialis anterior 91.4%, 11.5%, and 13.7% respectively in cancer cachexia mice. Proton NMR-based metabolomics revealed the altered metabolic profile and found 25 sensitive metabolites associated with cancer cachexia. Moreover, curcumin treatment resulted in metabolic reprogramming including decrease of phenylalanine, alanine, carnosine, carnitine, taurine, S-sulfocysteine, citrate, malate, glucose, and increase of citrulline, valine, isoleucine, methionine, glycine, acetoacetate and lactate. The pathway analysis showed that the main metabolic regulation of curcumin involved the metabolism of valine, leucine and phenylanaline, and synthesis and degradation of ketone bodies. Conclusions: these altered metabolic pathways imply a highly specific metabolism regulation of curcumin and raise the possibility for its therapeutic effect on alleviating cachexia hypermetabolism.

Selumetinib attenuate skeletal muscle wasting in murine cachexia model through ERK inhibition and AKT activation.

Cancer cachexia is a multifactorial syndrome affecting the skeletal muscle. Previous clinical trials showed that treatment with MEK inhibitor selumetinib resulted in skeletal muscle anabolism. However, it is conflicting that MAPK/ERK pathway controls the mass of the skeletal muscle. The current study investigated the therapeutic effect and mechanisms of selumetinib in amelioration of cancer cachexia. The classical cancer cachexia model was established via transplantation of CT26 colon adenocarcinoma cells into BALB/c mice. The effect of selumetinib on body weight, tumor growth, skeletal muscle, food intake, serum proinflammatory cytokines, E3 ligases, and MEK/ERK–related pathways was analyzed. Two independent experiments showed that 30 mg/kg/d selumetinib prevented the loss of body weight in murine cachexia mice. Muscle wasting was attenuated and the expression of E3 ligases, MuRF1 and Fbx32, was inhibited following selumetinib treatment of the gastrocnemius muscle. Furthermore, selumetinib efficiently reduced tumor burden without influencing the cancer cell proliferation, cumulative food intake, and serum cytokines. These results indicated that the role of selumetinib in attenuating muscle wasting was independent of cancer burden. Detailed analysis of the mechanism revealed AKT and mTOR were activated, while ERK, FoxO3a, and GSK3β were inhibited in the selumetinib -treated cachexia group. These indicated that selumetinib effectively prevented skeletal muscle wasting in cancer cachexia model through ERK inhibition and AKT activation in gastrocnemius muscle via cross-inhibition. The study not only elucidated the mechanism of MEK/ERK inhibition in skeletal muscle anabolism, but also validated selumetinib therapy as an effective intervention against cancer cachexia. Mol Cancer Ther; 16(2); 334–43. ©2016 AACR.

Protective Effects of Dexrazoxane against Doxorubicin-Induced Cardiotoxicity: A Metabolomic Study.

Cardioprotection of dexrazoxane (DZR) against doxorubicin (DOX)-induced cardiotoxicity is contentious and the indicator is controversial. A pairwise comparative metabolomics approach was used to delineate the potential metabolic processes in the present study. Ninety-six BALB/c mice were randomly divided into two supergroups: tumor and control groups. Each supergroup was divided into control, DOX, DZR, and DOX plus DZR treatment groups. DOX treatment resulted in a steady increase in 5-hydroxylysine, 2-hydroxybutyrate, 2-oxoglutarate, 3-hydroxybutyrate, and decrease in glucose, glutamate, cysteine, acetone, methionine, asparate, isoleucine, and glycylproline.DZR treatment led to increase in lactate, 3-hydroxybutyrate, glutamate, alanine, and decrease in glucose, trimethylamine N-oxide and carnosine levels. These metabolites represent potential biomarkers for early prediction of cardiotoxicity of DOX and the cardioprotective evaluation of DZR.

Steady-state plasma concentration of donepezil enantiomers and its stereoselective metabolism and transport in vitro.

The aim of the present study was to elucidate the differences in the plasma concentration of two enantiomers of donepezil in Chinese patients with Alzheimers disease (AD) and investigate in vitro stereoselective metabolism and transport. Donepezil enantiomers were separated and determined by LC-MS/MS using D5-donepezil as an internal standard on a Sepax Chiralomix SB-5 column. In vitro stereoselective metabolism and transport of donepezil were investigated in human liver microsomes and MDCKII-MDR1 cell monolayer. Pre-dose (Css-min) plasma concentrations were determined in 52 patients. The mean plasma level of (R)-donepezil was 14.94 ng/ml and that of (S)-donepezil was 23.37 ng/ml. One patients plasma concentration of (R)-donepezil was higher than (S)-donepezil and the ratio is 1.51. The mean plasma levels of (S)-donepezil were found to be higher than those of (R)-donepezil in 51 patients and the ratio of plasma (R)- to (S)-donepezil varies from 0.34 to 0.85. In the in vitro microsomal system, (R)-donepezil degraded faster than (S)-donepezil. V(max) of (R)-donepezil was significantly higher than (S)-donepezil. The P-gp inhibition experiment shown that the P(app) of the two enantiomers was higher than 200 and the efflux ratios were 1.11 and 0.99. The results of the P-gp inhibition identification experiment showed IC50 values of 35.5 and 20.4 μM, respectively, for the two enantiomers. The results indicate that donepezil exhibits stereoselective hepatic metabolism that may explain the differences in the steady-state plasma concentrations observed. Neither (R)- nor (S)-donepezil was a P-gp substance and the two enantiomers are highly permeable through the blood-brain barrier.

Distinct Metabolic Profile of Inhaled Budesonide and Salbutamol in Asthmatic Children during Acute Exacerbation

Inhaled budesonide and salbutamol represent the most important and frequently used drugs in asthmatic children during acute exacerbation. However, there is still no consensus about their resulting metabolic derangements; thus, this study was conducted to determine the distinct metabolic profiles of these two drugs. A total of 69 children with asthma during acute exacerbation were included, and their serum and urine were investigated using high‐resolution nuclear magnetic resonance (NMR). A metabolomics analysis was performed using a principal component analysis and orthogonal signal correction–partial least squares using SIMCA‐P. The different metabolites were identified, and the distinct metabolic profiles were analysed using MetPA. A high‐resolution NMR‐based serum and urine metabolomics approach was established to study the overall metabolic changes after inhaled budesonide and salbutamol in asthmatic children during acute exacerbation. The perturbed metabolites included 22 different metabolites in the serum and 21 metabolites in the urine. Based on an integrated analysis, the changed metabolites included the following: increased 4‐hydroxybutyrate, lactate, cis‐aconitate, 5‐hydroxyindoleacetate, taurine, trans‐4‐hydroxy‐l‐proline, tiglylglycine, 3‐hydroxybutyrate, 3‐methylhistidine, glucose, cis‐aconitate, 2‐deoxyinosine and 2‐aminoadipate; and decreased alanine, glycerol, arginine, glycylproline, 2‐hydroxy‐3‐methylvalerate, creatine, citrulline, glutamate, asparagine, 2‐hydroxyvalerate, citrate, homoserine, histamine, sn‐glycero‐3‐phosphocholine, sarcosine, ornithine, creatinine, glycine, isoleucine and trimethylamine N‐oxide. The MetPA analysis revealed seven involved metabolic pathways: arginine and proline metabolism; taurine and hypotaurine metabolism; glycine, serine and threonine metabolism; glyoxylate and dicarboxylate metabolism; methane metabolism; citrate cycle; and pyruvate metabolism. The perturbed metabolic profiles suggest potential metabolic reprogramming associated with a combination treatment of inhaled budesonide and salbutamol in asthmatic children.

Integrated analysis of serum and intact muscle metabonomics identify metabolic profiles of cancer cachexia in a dynamic mouse model

Cancer cachexia is a multifactorial metabolic syndrome characterized by a severe loss of body weight and lean body mass. Metabolic dysfunction is the primary hallmark of muscle atrophy. Herein, we studied dynamic metabolic profiles in serum and intact muscle. High-resolution magic angle spinning was employed for intact gastrocnemius muscle analysis and a dynamic metabolic model was established using C26 colon carcinoma-bearing mice from procachexia to the refractory cachexia period. When an integrated analysis of the 13 metabolites from the intact muscle gastrocnemius and 43 metabolites from the serum was performed, five distinguishable metabolic features were identified, including low blood glucose, elevated ketone bodies, decreased branched-chain amino acids, increased neutral amino acids, and high 3-methylhistidine and creatine. The metabolic hubs reveal potential biomarkers for the early detection of cachexia and indicate the underlying metabolic pathway reprogramming of muscle atrophy.