Synthesis and biological evaluation of sophocarpinic acid derivatives as anti-HCV agents

Chronic hepatitis C virus(HCV)infection has become a major public health burden worldwide.Twenty-two sophocarpinic acid or matrine derivatives were synthesized and their anti-HCV activities were evaluated in vitro.The structure-activity analysis revealed that(i)sophocarpinic acids with a D-seco 3-ring structure scaffold were more favorable than matrines with a 4-ring scaffold;(ii)the introduction of an electron-withdrawing group on the phenyl ring in 12-N-benzenesulfonylΔβγsophocarpinic acids was beneficial for the antiviral activity against HCV.Among them,compounds 9h and 9j exhibited the most potent inhibitory activities on HCV replication with selectivity indies of 70.3 and 30.9,respectively.Therefore,both were selected as antiviral candidates for further investigation.

Quality Standard of Qiang Herb Sophora davidii var.chuansiensis

[Objectives]To establish the quality standard of common clinical folk herb Sophora davidii var.chuansiensis.[Methods]The qualitative identification was performed by microscopic method and thin-layer chromatography(TLC).The contents of moisture,total ash,acid insoluble ash and extract were determined according to the methods of the Chinese Pharmacopoeia(2020).High performance liquid chromatography(HPLC)was used to determine the contents of oxymatrine and oxysophocarpine.[Results]The microscopic characteristics were obvious,including crystal sheath fiber,calcium oxalate square crystal,non glandular hair,stone cells,epidermal cells,stomata,cork cells,vessels with marginal pits and so on.TLC spots were clear,and the resolution was good.The contents of moisture,total ash,acid insoluble ash and extract from 10 batches of samples were 4.70%-8.33%,3.43%-4.19%,0.65%-1.02%and 14.67%-22.04%respectively.The determination results of oxymatrine were between 0.10%-0.33%,with an average value of 0.19%.The determination results of sophocarpine oxide were between 0.30%-0.38%,with an average value of 0.34%.[Conclusions]The established quality standard of S.davidii var.chuansiensis had good specificity and accuracy,and could be used for the quality control of herb S.davidii var.chuansiensis.

Sophocarpine attenuates liver fibrosis by inhibiting the TLR4 signaling pathway in rats

AIM:To explore the effect of sophocarpine on experimental liver fibrosis and the potential mechanism involved.METHODS:Sophocarpine was injected intraperitoneally in two distinct rat hepatic fibrosis models induced either by dimethylnitrosamine or bile duct ligation.Masson’s trichrome staining,Sirius red staining and hepatic hydroxyproline level were used for collagen determination.Primary hepatic stellate cells(HSCs)were isolated and treated with different concentrations of sophocarpine.Real-time reverse transcription-polymerase chain reaction was used to detect the mRNA levels of fibrotic markers and cytokines.The expression of pathway proteins was measured by Western blot.The Cell Counting Kit-8 test was used to detect the proliferation rate of activated HSCs treated with a gradient concentration of sophocarpine.RESULTS:Sophocarpine decreased serum levels of aminotransferases and total bilirubin in rats under chronic insult.Moreover,administration of sophocarpine suppressed extracellular matrix deposition and prevented the development of hepatic fibrosis.Furthermore,sophocarpine inhibited the expression ofα-smooth muscle actin(SMA),interleukin(IL)-6,transforming growth factor-β1(TGF-β1),Toll-like receptor 4(TLR4),and extracellular-related kinase(ERK)in rats.Sophocarpine also down-regulated the mRNA expression ofα-SMA,collagenⅠ,collagenⅢ,TGF-β1,IL-6,tumor necrosis factor-αand monocyte chemoattractant protein-1,and decreased protein levels of TLR4,p-ERK,p-JNK,p-P38 and p-IKK in vitro after Lipopolysaccharide induction.In addition,sophocarpine inhibited the proliferation of HSCs accompanied by a decrease in the expression of Cyclin D1.The protein level of proliferating cell nuclear antigen was decreased in activated HSCs following a gradient concentration of sophocarpine.CONCLUSION:Sophocarpine can alleviate liver fibrosis mainly by inhibiting the TLR4 pathway.Sophocarpine may be a potential chemotherapeutic agent for chronic liver diseases.

Protection of Sophocarpine on Colonic Barrier in DSS-induced Acute Colitis in Mice by Increasing Expression of HNF4α

Objective To investigate the possible protective effects of sophocarpine on mucosal injury and epithelial barrier disruption on dextran sulfate sodium （DSS）-induced acute colitis. Methods Male BALB/c mice were randomly divided into three groups. The mice in normal group were given normal water, and those in model and sophocarpine- treated groups were given 2.5% DSS for 6 d to induce acute colitis. Sophocarpine （30 mg/kg） was ip administered once daily during the study period. Severity of colitis was evaluated by disease activity index （DAI）, histological injury and inflammatory cytokine production including tumor necrosis factor-α （TNF-α）, interleukin-lβ （IL-Iβ）, and monocyte chemoattractant protein-1 （MCP-1）. The colonic barrier disruption was assessed by testing the expression of junctional adhesion molecule-1 （JAM-1）, E-cadherin （E-CAD）, and desmocollin-2 （DSC-2） in colon mucosa. Expression of HNF4~ in colon mucosa was detected by immunohistochemistry staining and real-time RT-PCR, respectively. Results Compared with normal group, DAI, colonic shortening, and histopathological injury in model group were elevated （P 〈 0.05）, but reduced in sophocarpine-treated group （P 〈 0.05）. Compared with model group, the mRNA expression of inflammatory cytokines （TNF-α, IL-16, MCP-1） were obviously lower in sophocarpine-treated group （P 〈 0.05）, while the cellular junction proteins （E-CAD, JAM-l, and DSC-2） were higher （P 〈 0.05）. The expression of HNF4α at mRNA and protein levels was decreased significantly in model group, but increased apparently in sophocarpine-treated group. Conclusion Sophocarpine can enhance the expression of HNF4α, promote the expression of colonic intrecellular junctions, thus, maintain the integrity of the colonic barrier and inhibit the colitis process. We suggest that sophocarpine could enhance the production of cellular junction proteins to protect the intestinal barrier fuction, at least partly, in HNF4α-dependent pathway.