Sex-related differences in safety profiles, pharmacokinetics and tissue distribution of sinomenine hydrochloride in rats.

Sinomenine is a bioactive alkaloid isolated from the Chinese medicinal plant Sinomenium acutum (Thunb.) Rehd. et Wils which exhibits significant analgesic, anti-inflammatory, and immunosuppressive effects. Sinomenine hydrochloride (SH) preparations, classified as natural disease-modifying antirheumatic drugs, are currently available for the treatment of rheumatoid arthritis and other rheumatic diseases. Our toxicity evaluation demonstrated that the median lethal dose of SH in female Sprague-Dawley (SD) rats was over 11 times greater than that in male SD rats, revealing striking sex-linked differences in the safety profile of SH. The present study was designed to investigate differences in the pharmacokinetics (PKs) and tissue distribution of SH between male and female SD rats after a single oral dose of 25 mg/kg. PK and tissue distribution studies were performed using a validated UPLC-MS/MS method. The results showed that SH-treated SD female rats displayed markedly greater drug exposure, and SH exhibited a longer half-life and slower clearance rate than comparable studies in male rats. Moreover, the tissue distribution study confirmed that the sinomenine concentration in female rats was considerably greater in the internal organs than in male rats. Our study demonstrates, for the first time, significant sex-related differences in the safety profile and PKs of SH, which may be associated with a distinct sex-dependent metabolic mechanism of sinomenine.

Role of Postoperative Radiotherapy in Nonmetastatic Head and Neck Adenoid Cystic Carcinoma.

BACKGROUND: Head and neck adenoid cystic carcinoma (ACC) is a rare malignant tumor that is prone to local recurrence. The NCCN Guidelines for Head and Neck Cancers recommend that all patients with ACC receive postoperative radiotherapy (PORT). However, whether PORT can improve local control and which patients can benefit from PORT are unknown. This study aimed to assess the role of PORT and provide individualized suggestions for postoperative therapy in patients with ACC. PATIENTS AND METHODS: We retrospectively reviewed patients with nonmetastatic head and neck ACC who underwent surgery with or without PORT. Recursive partitioning analysis (RPA) was performed to categorize the patients and predict local recurrence-free survival (LRFS). The survival outcome was compared between non-PORT and PORT groups. RESULTS: A total of 319 patients were included. PORT was identified as a prognostic factor for LRFS in univariate (P=.01) and multivariate analysis (P<.01). However, it did not improve distant metastasis-free survival, disease-free survival, or overall survival in univariate analysis. RPA categorized patients into 3 prognostic groups: low-risk (negative margin, T1-T2, primary location = major or minor salivary gland), intermediate-risk (negative margin, T1-T2, primary location = other locations instead of a major or minor salivary gland; negative margin, T3-T4; positive margin, without bone invasion), and high-risk (positive margin, with bone invasion). Significant LRFS improvements in the PORT group were observed among intermediate-risk (P<.01) and high-risk patients (P<.05). LRFS improvements among low-risk patients were relatively insignificant (P=.10). CONCLUSIONS: PORT was shown to be a positive prognostic factor for improved LRFS in ACC. Furthermore, PORT could significantly improve LRFS in intermediate-risk and high-risk patients with ACC, but whether low-risk patients could benefit from PORT needs further study.

Nitrogen-containing flavonoid and their analogs with diverse B-ring in acetylcholinesterase and butyrylcholinesterase inhibition.

In this study, a series of new flavones (2-phenyl-chromone), 2-naphthyl chromone, 2-anthryl-chromone, or 2-biphenyl-chromone derivatives containing 6 or 7-substituted tertiary amine side chain were designed, synthesized, and evaluated in acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition. The results indicated that the alteration of aromatic ring connecting to chromone scaffold brings about a significant impact on biological activity. Compared with flavones, the inhibitory activity of 2-naphthyl chromone, 2-anthryl-chromone derivatives against AChE significantly decreased, while that of 2-biphenyl chromone derivatives with 7-substituted tertiary amine side chain is better than relative flavones derivatives. For all new synthesized compounds, the position of tertiary amine side chain obviously influenced the activity of inhibiting AChE. The results above provide great worthy information for the further development of new AChE inhibitors. Among the newly synthesized compounds, compound 5a is potent in AChE inhibition (IC50 = 1.29 ± 0.10 µmol/L) with high selectivity for AChE over BChE (selectivity ratio: 27.96). An enzyme kinetic study of compound 5a suggests that it produces a mixed-type inhibitory effect against AChE.

Structure-activity relationship investigation of coumarin-chalcone hybrids with diverse side-chains as acetylcholinesterase and butyrylcholinesterase inhibitors.

Chalcones containing tertiary amine side-chains have potent activity as acetylcholinesterase (AChE) inhibitors. However, the effects of the location of the tertiary amine groups as well as of other groups on AChE and butyrylcholinesterase (BChE) activity have not been reported. Here, we report the synthesis and testing of 36 new coumarin-chalcone hybrids (5d-7j, 9d-11f, 12k-13m) against AChE and BChE. The nature and position of the chalcone substituents had major effects on inhibitory activity as well as selectivity for AChE over BChE. Compounds with para-substituted chalcone fragments in which the substituents were choline-like had potent activity against AChE and poor activity against BChE, while ortho-substituted analogs exhibited an opposite effect. Replacement of the terminal amine groups by amide, alkyl or alkenyl groups abrogated activity. Compound 5e showed potent inhibitory activity [Formula: see text]) and good selectivity for AChE over BChE (ratio 27.4), and a kinetic study showed that 5e exhibited mixed-type inhibition against AChE. Computational docking results indicate that 5e binds to Trp 279, Tyr334 and Trp 84 in AChE, but only to Trp 82 in BChE. Overall, the results show that coumarin-chalcone hybrids with choline-like side-chains have promising activity and selectivity against AChE and be promising therapeutic leads for Alzheimer's disease.

[Analytical Methods with Qualitative HPLC Fingerprint and Quantitative Measurement of Effective Components of Processed Asari Radix et Rhizoma].

OBJECTIVE: In order to provide the basis for quality standard and processing principle of processed Asari Radix et Rhizoma, an HPLC fingerprint of processed Asari Radix et Rhizoma was established and the contents of methyl eugenol and asarinin were determined. METHODS: The analytical column was Agligent Tc-C18 (250 mm x 4. 6 mm, 5 µm); A mixture of acetonitrile-0. 2% acetic acid solution was used as the mobile phase with gradient elution at the flow rate of 1. 0 mL/min. The wavelength was set at 285 nm and the column temperature was 30 °C. RESULTS: The fingerprint of processed Asari Radix et Rhizoma was established. The asarinin peak was taken as the reference peak. 22 common peaks were assigned, and two peaks were confirmed by comparing with the reference standards. The difference of components was not significant among the various processed products except ginger, honey and fried coke products, but the contents of effective constituents were different among the processed products. The retention rate of methyl eugenol in processed products was in a descending order as follows: wine > vinegar > liquorice > alkali-vinegar > fried coke > rice water system > honey > ginger > salt system > alkali. Methyl eugenol was increased 10% ~ 20% with wine processing and retained more than 95% with vinegar. The retention rate of asarinin in processed products was in declining as: rice water system > liquorice > alkali-vinegar > honey > salt system > wine > ginger > vinegar > alkali > fried coke. The processing techniques increased the content of asarinin except the alkali and fried coke products, and asarinin was increased more than 35% with rice water, alkali-vinegar or liquorice processing. CONCLUSION: The method is accurate and reliable, which can be used for the quality control of processed products of Asari Radix et Rhizoma.

[Study on pharmacokinetics-pharmacodynamics correlation of yin teng gu bi kang prescription].

OBJECTIVE: To study pharmacokinetics-pharmacodynamics (PK-PD) correlation of Yin Teng Gu Bi Kang (YTGBK) prescription through determination of Tanshinone II(A) concentration and the level of Malondialdehyde (MDA) in plasma in normal and blood stasis rats treated with YTGBK prescription. METHODS: The concentration of Tanshinone II(A) in the plasma was measured by HPLC-UV and loratadine was used as internal standard; Thiobarbituric acid reactive substance assay (TBARS) was adopted to determine the concentration of MDA in the plasma Area under the concentration-time curve (AUC) and area under the effect-time curve (AUE) were calculated using linear trapezoid rule. The correlation and regression analysis was performed by plotting AUE (Y) versus lgAUC (X) using linear regression. RESULTS: YTGBK prescription could significantly decrease MDA level in the plasma in above two different physiological rats at the analyzed time point (P < 0.05). Scatter plots of AUE-lgAUC showed an upward trend. The results of the correlation and regression analysis were as follows: Y = 53.367 X -30.780, r = 0. 822, P = 0.007 for normal rats and Y = 61.091 X -39.863, r = 0.777, P = 0.003 for model rats, respectively. CONCLUSION: There is a positive correlation between Tanshinone II(A) level in plasma and the antioxidant activity of YTGBK prescription in decreasing MDA level, which indicates that Tanshinone II(A) is the antioxidant effective substance of YTGBK prescription.

[Study on material basis of essential oil from Yin Teng Gu Bi Kang prescription on activating blood circulation].

OBJECTIVE: To explore the component difference of the serum containing essential oil from Yin Teng Gu Bi Kang prescription in pathologic and physiologic rat models, and to reveal the material basis of its efficacy of activating blood circulation. METHODS: The essential oils were obtained by CO2 supercritical fluid extraction and the ingredients of the essential oils in vitro and in vivo (under physiological and pathological status) were analyzed by GC-MS to compare differences of the essential oil under physiological and pathological status in rats. RESULTS: 32 components were identified with the main components of Z-ligustilide (39.23%) and d-limonene (21.7%) in the essential oil. In vivo analysis on the essential oil indicated that 16 components were identified, 7 existed originally in essential oil and 9 were metabolites under physiological status; while 22 components were identified, 10 existed originally in essential oil and 12 were metabolites under pathological status (acute blood stasis). There were 7 common prototypes and 8 common metabolites under different physiological status. CONCLUSION: The absorption and metabolism of essential oils were affected by blood stasis and the compounds migrating to blood may be the effective substance in activating blood circulation.

[Comparison of pharmaceutical chemistry difference of effective parts from yin teng gu bi kang prescription in normal rats and rats with acute blood stasis].

OBJECTIVE: To elucidate the material basis of Yin Teng Gu Bi Kang Prescription (YTGBKP) for efficacy of promoting blood circulation by means of comparing the pharmaceutical chemistry difference of effective parts in normal rats and rats with acute blood stasis. METHODS: The pharmaceutical chemistry fingerprints of effective parts under physiological and pathological status (acute blood stasis) were established by HPLC,and the in vitro and in vivo chromatographic peaks were compared and analyzed. RESULTS: Five batches of drug-containing plasma samples had 14 chromatographic peaks under normal physiological status,among which 3 rooted in plasma, 9 existed originally in YTGBKP,2 were metabolites. The compound with retention lime at 12 min was identified as ferulic acid by comparing with reference standard; While under pathological status (acute blood stasis), five batches of the drug-containing plasma samples had 14 chromatographic peaks, among which 3 rooted in plasma, 9 existed originally in YTGBKP, 2 were metabolites. The compounds with retention time at 12 min and 32 min were identified as ferulic acid and icariin respectively by comparing with reference standards. There were 10 common peaks under normal physiological and pathological status (acute bloodl stasis) excluding peaks in blank plasma. The intensity of the common peaks produced under pathological status was stronger than that under normal physiological status significantly; Variance analysis showed that there were significant differences (P < 0.05) in peak areas of 3 peaks. CONCLUSION: Blood stasis has influence on the absorption and metabolism of most ingredients from YTGBKPi; Prototypes and metabolites may be the effective substance on promoting blood circulation.

Celastrol ameliorates vascular neointimal hyperplasia through Wnt5a-involved autophagy.

Neointimal hyperplasia caused by the excessive proliferation of vascular smooth muscle cells (VSMCs) is the pathological basis of restenosis. However, there are few effective strategies to prevent restenosis. Celastrol, a pentacyclic triterpene, has been recently documented to be beneficial to certain cardiovascular diseases. Based on its significant effect on autophagy, we proposed that celastrol could attenuate restenosis through enhancing autophagy of VSMCs. In the present study, we found that celastrol effectively inhibited the intimal hyperplasia and hyperproliferation of VSMCs by inducing autophagy. It was revealed that autophagy promoted by celastrol could induce the lysosomal degradation of c-MYC, which might be a possible mechanism contributing to the reduction of VSMCs proliferation. The Wnt5a/PKC/mTOR signaling pathway was found to be an underlying mechanism for celastrol to induce autophagy and inhibit the VSMCs proliferation. These observations indicate that celastrol may be a novel drug with a great potential to prevent restenosis.