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Bicyclic alcohol on immune hepatic fibrosis in rats and its mechanism research oral 2.Beagle dog fumarate tenofovir topiramate furosemide esters and BP0018 plasma pharmacokinetics and bioavailability studies

Author: GuYu
Tutor: GuoShunXing；LiYan；ChengGuiFang；ChenXiaoGuang；ShenZhuFang
School: Peking Union Medical College , China
Course: Pharmacognosy
Keywords: Liver Fibrosis Bovine serum albumin (BSA) Bicyclol Cytokines Smad 2 / 3 p38 MAPK MMP-2 TIMP-1 BP0018 The fumarate tenofovir dipyrazole , furosemide esters Tenofovir Anti - virus Pharmacokinetics Bioavailability
CLC: R965
Type: PhD thesis
Year: 2010
Downloads: 295
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Abstract

The first part of the bicyclol protective effect on immune hepatic fibrosis in rats and mechanism of liver fibrosis (liver fibrosis,) liver cell necrosis and inflammation the liver abnormal proliferation of fibrous connective tissue in the pathological process, not only is One of the most important pathological features of chronic liver disease, also chronic hepatitis, cirrhosis and further development of the deterioration of the important reasons. Although liver fibrosis is reversible pathological process, but if not timely and effective treatment, and ultimately may develop irreversible liver cirrhosis and even liver cancer. Therefore, the study of the formation mechanism of liver fibrosis and the prevention and treatment of liver fibrosis has become one of the hot topics in the world of medicine and pharmacology research. Is used to study the mechanism of liver fibrosis commonly used animal model of including CCl4, two subunit nitrosamines, bile duct ligation, immune hepatic fibrosis model, in which the immune hepatic fibrosis model with clinical chronic hepatitis due to hepatic fibrosis of closer in the pathogenesis of immune mechanisms of the characteristics of the study for chronic hepatitis. Bicyclol anti-hepatitis drug developed by the Chinese Academy of Medical Sciences, has been used in the treatment of chronic viral hepatitis. Clinical data show that the bicyclic alcohol can significantly improve chronic hepatitis B, hepatitis C patients with clinical symptoms and liver injury, both must inhibit the replication of hepatitis B virus, but also has low rebound, adverse reactions taking convenient features. Previous studies show that bicyclol on a variety of experimental liver injury have a significant protective effect, protection mechanisms and scavenging reactive oxygen free radicals, regulation of cytokine secretion, inhibition of immune injury-induced apoptosis. In this paper, bovine serum albumin-induced rat immune hepatic fibrosis model Learn more about bicyclol features a protective effect on liver fibrosis, and nuclear transcription factors, cytokines, regulation explore bicyclol protective effect mechanism for the clinical application to provide scientific and reliable experimental basis. The occurrence mechanism bicyclol protective effect on the autoimmune liver fibrosis liver injury in rats and mechanism of bovine serum albumin (Bovine Serum Albumin, BSA) induced rat liver fibrosis model is based on heterologous serum albumin into the body can stimulate body to produce antibodies, followed by the formation of immune complexes activate complement, triggering a type III hypersensitivity reaction caused by the the vasculitis, perivascular go far, local immune and inflammatory response, further stimulate the proliferation of collagen and the formation of liver fibrosis. In this study, the BSA-induced liver fibrosis in rats visible elevated levels of liver hydroxyproline (Hydroxyproline, Hyp), hyaluronic acid (Hyaluronic Acid HA) and III procollagen peptide (Procollagen Type III, P III P) content a significant increase in interstitial collagen fibers in the liver and an increase in collagen, reticular fibers and collagen fibers hyperplasia periportal fibrosis pathological changes, in line with previous studies. Administration group at the start of the attack phase four weeks after the start giving bicyclol (100, 200, 300 mg / ml) once a day for 5 wks after administration can significantly reduce the BSA-induced liver fibrosis in rat liver HYP, serum HA and PIIIP level rise, the infiltration of inflammatory cells, fibrous tissue, and liver pathological changes also significantly reduced. Regulation of the promoter activity of the known process of liver fibrosis and liver extracellular matrix deposition adds and changes are closely related to the main features of the hepatic stellate cell activation, transforming growth factor (Transforming growth factorβ-1, TGF-β1) and its downstream cell changed. TGF-beta1 can regulate extracellular matrix (Extracellularmatrix ECM) the mesh system components such as fiber collagen protein and fiber links, as well as protease inhibitors, including matrix metalloproteinase inhibitors (Tissue inhibitor of metalloproteinase, TIMPs) to adjust. Previous studies have shown that TGF-beta1 signal transduction pathways prevention has become an important target for the treatment of liver fibrosis. The results of this study show that bicyclol inhibition of hepatic overexpression of TGF-beta1 mRNA and protein levels significantly, suggesting bicyclol thereby preventing the formation of liver fibrosis by down-regulating hepatic TGF-beta1. Liver damage TGF-beta-through the intracellular TGF-β/Smad signal pathway and accelerate hepatic stellate cells to differentiate into myofibroblasts. Addition, TGF-β/Smad passage may also interact with other signaling pathways, to increase or inhibit the phosphorylation of Smad proteins, of which the most important is activated protein kinase pathway (MAPK) and p38 MAPK. Studies have demonstrated Smad3 phosphorylation-dependent p38 MAPK, to promote myofibroblast in vitro and in vivo generation of ECM, P38 MAPK multiple stimuli can be various MAPKKKs kinase activation, and thus at different levels to promote myoblast differentiation . The visible of Smad2 / 3 phosphorylation level of bicyclic alcohols administration group was significantly reduced, and p38 MAPK phosphorylation was inhibited due, at least prompt bicyclol inhibition of the expression of TGF-beta and Smad2 / 3 and p38 MAPK signal pathways related to the regulation. IL-1 is one of the most important inflammatory cytokines by promoting liver fibrosis mechanism mainly includes the following two aspects: 1) the inflammatory response generated by IL-1 in liver tissue can be activated p38, JNK, resulting in hepatic fibrosis generated ; 2) IL-1 to increase TIMP-1 mRNA expression, and promote ECM accumulation and liver fibrosis. Seen in this study to BSA induced plasma interleukin prime-1beta (Interleukin-1beta, IL-1beta) mRNA levels were significantly increased, bicyclic alcohol can inhibit IL-1beta increased, may bicyclol, by inhibiting p38MAPK thereby reducing the inflammatory response of the liver relevant. In addition, in the process of liver fibrosis, TNF-alpha on the proliferation of HSC activation and ECM synthesis, matrix metalloproteinases, and tissue inhibitor of release plays an important role. BSA-induced rat liver fibrosis induced TNF-alpha mRNA levels were significantly increased, bicyclol can significantly reduce its expression. Interleukin -10 (Interleukin-10, IL-10) is important in vivo anti-inflammatory cytokine binding to its receptor can inhibit a variety of inflammatory cytokines and pro-fibrotic factors including IL-1beta. The liver immune inflammatory process seen in the Kupffer cells, HSCs and hepatic infiltration of T cells in IL-10 mRNA expression. Previous studies have shown that exogenous IL-10 can antagonize CCl4-induced hepatic fibrosis in rats, and can inhibit the in vitro the culture activation HSCTGF-beta 1 and basic fibroblast growth factor mRNA expression and promote apoptosis of HSC. This study found that the BSA attack rat liver IL-10 reactivity increased, while the the bicyclic alcohol may reduce the over-expression of IL-10. Speculate because bicyclol significantly down-regulated the expression of pro-inflammatory cytokines in the liver, inhibit liver inflammation, feedback reduces the increase in IL-10. Previous studies have shown that liver fibrosis deposition and degradation of collagen and matrix metalloproteinase (Matrix metalloproteinase, MMPs) and TIMPs is closely related. In a healthy liver, the the ECM steady-state environment by MMPs guidance precise regulation. In chronic liver injury, hepatic stellate cells are activated and differentiate into fibroblast-like phenotype. At the same time, the increase in the inhibition of MMP activity and subsequent extracellular space matrix protein accumulation in activated HSCs in the expression of TIMP-1. The visible bicyclol BSA-induced MMP-2 and TIMP-1 mRNA overexpression significantly reduced. Although not yet determine whether the increase in collagen activity is bicyclol inhibit the expression of TIMP-1, TIMP-1 reduction and increased collagenase activity is often associated with the reversal of liver fibrosis. Reports indicate that IL-1 activation of p38 MAPK can lead to upregulation of TIMP-1. The above results, the the bicyclic alcohol may inhibit the expression of inflammatory cytokines (inhibition of IL-1beta), inhibition of phosphorylation of p38MAPK, and interfere with the TIMP-1 thereby inhibiting the expression of collagen, to prevent the occurrence of liver fibrosis by regulating TGF-beta1. Thus, the role of the mechanisms to prevent the occurrence of inflammation reactions, so that the level of IL-10 reduced feedback Meanwhile, the bicyclic alcohol treatment can significantly reduce the expression of MMP-2 and TIMP-1, the analysis is by inhibition of the p38 MAPK phosphorylation, interference TIMP-1 inhibition of collagen expression, and ultimately prevent the occurrence of liver fibrosis. In summary, bicyclol BSA-induced liver fibrosis has a protective effect and its mechanism of action can be summarized as: 1) inhibition of hepatic cells through Smad2 / 3 and p38 MAPK signal pathway regulating TGF-beta1 and IL-1beta expression, deposition of extracellular matrix; 2) inhibition of inflammatory cytokines TNF-alpha and IL-1 expression, thereby improving the inflammatory factor / anti-inflammatory cytokine balance; 3) through the regulation of MMP-2 and TIMP-1 inhibition of collagen expression and treatment of liver fibrosis. Learn more about the research of bicyclol liver protective role of the characteristics used in clinical treatment of liver fibrosis provide the experimental basis of the reference value. The second part of the Beagle dog oral fumarate the tenofovir dipyrazole furosemide esters and BP0018 plasma pharmacokinetics Pharmacokinetics and bioavailability degree of liver disease, chronic hepatitis B, HBsAg to carry the number of the total population has reached 10 %, an annual increase of about 50 million hepatitis B patients, the number of people die each year from hepatitis B is about 28 million. At the same time, according to the latest statistics, the world's 25 million people died of HIV infection, HIV infection there are 33 million people in our country, existing HIV virus infection and the patient has more than 70 million people. Anti-HIV and anti-HBV drug development for the treatment of viral infection has great social and economic significance. Viread a highly resistant HBV and HIV activity of the novel nucleoside reverse transcriptase inhibitor, has been approved in the United States and Europe, the treatment of human immunodeficiency virus (HIV) infection, and is also an effective drug for anti-chronic hepatitis B drug, by interfering with the function of the hepatitis B virus DNA polymerase, inhibiting the replication of hepatitis B virus, and reduce the viral load in the serum and liver tissue. BP0018 is the Pharmaceutical JCTT developed anti-viral drugs, Viread, are prodrug of tenofovir (PMPA). The paper aims to study the Beagle dogs single oral dose viread and BP0018 plasma drug pharmacokinetic characteristics, and calculate the bioavailability, provide the basis for the BP0018 clinical rational drug safety and efficacy. This study is based chemical drugs bioavailability and bioequivalence studies technical guiding principles, a single oral dose of Beagle dogs BP0018 and reference vivo plasma pharmacokinetics of the preparation viread dynamics and biological utilization study. The results show that: (1) established in this study for the determination of biological samples Tenofovir, BP0018 and tenofovir topiramate furosemide esters HPLC-MS/MS method free from impurities interference, intra-day precision RSD was less than 2.8%, day RSD less than 10.1%, the recovery rate in the range of 80.9 to 93.1%, RSD were less than 8.2%, and stability to meet the in vivo pharmacokinetic studies requirements. (2) Beagle dogs single oral dose of BP0018 and reference formulation prototype drug concentration at each time point were lower than the limit of detection (5ng/mL), but can be detected in the blood to tenofovir (PMPA), its concentration with at different times after administration changes; (3) BP0018 and the reference preparation PMPA peak time were 0.25 h and 0.25-2H, peak concentration were 328.06-65.74ng/mL and 331.21 ± 22.74ng/mL of body mean residence time (MRT) were 6.87 ± 1.52h and 4.35 ± 1.61h; (4) and compared to the reference formulation BP0018 mean residence time (6.876.87 ± 1.52h) and elimination half-life (14.95 ± 9.49h) are extended, and AUC (0-t), AUC (0 - ∞), Cmax and Tmax was no significant difference. (5) Beagle dogs after a single intravenous injection of PMPA, the plasma drug concentration after administration a 2min rapid peak 24h still detects a certain concentration of PMPA and the 36h plasma concentrations below the detection limit PMPA. (6) the in vivo bioavailability of the Beagle dog a single oral administration BP0018 to 20.7 ± 4.3%, and the bioavailability was 16.8 ± 6.2% fumaric acid the tenofovir dipyrazole furosemide esters reference preparation. Summary, this paper Beagle dogs single oral dose of tenofovir fumarate two topiramate furosemide esters and BP0018 plasma drug pharmacokinetic characteristics, and calculate its bioavailability, the results showed the two drugs drugs no significant difference in pharmacokinetic parameters, the bioavailability results suggest that the basic equivalent, to provide experimental basis for the BP0018 clinical rational drug safety and efficacy.

Bicyclic alcohol on immune hepatic fibrosis in rats and its mechanism research oral 2.Beagle dog fumarate tenofovir topiramate furosemide esters and BP0018 plasma pharmacokinetics and bioavailability studies

Abstract

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