Comprehensive Physicochemical and Biological Characterization of the Proposed Biosimilar Darbepoetin Alfa, LBDE, and Its Originator Darbepoetin Alfa, NESP®.

BACKGROUND: For regulatory approval, the comparability of a biosimilar product to an originator product should be ensured through thorough physicochemical and biological characterization. OBJECTIVE: To evaluate the biosimilarity between LBDE, the proposed biosimilar darbepoetin alfa, and NESP®, its originator, we performed a comprehensive physicochemical and biological characterization study. METHODS: Primary and higher-order protein structures were analyzed using Lys-C peptide mapping with liquid chromatography-mass spectrometry (LC-MS), disulfide bond identification, circular dichroism, and fluorescence spectroscopy. Glycosylation and isoform distribution were analyzed using MS, LC, and capillary zone electrophoresis. Size variants were evaluated with size-exclusion chromatography-high-performance liquid chromatography (SEC-HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Biological characterization included binding affinity for human erythropoietin receptor, in vitro cell proliferation, and in vivo potency. Pharmacokinetics (PK) were evaluated using rats through two injection routes. RESULTS: Non-reducing and reducing Lys-C peptide mapping showed a highly similar peak profile, confirming that LBDE and NESP® have the same primary structure and disulfide bonds. Glycosylation and isoform analyses showed that the attached N-glycan and O-glycan structures were the same and their relative contents were similar. Spectroscopic analysis of LBDE showed indistinguishable spectra with NESP®. For both LBDE and NESP®, a very small amount of size variants was found in SEC-HPLC, and no minor bands were detected in SDS-PAGE. Furthermore, LBDE did not show any difference with NESP® in the in vitro and in vivo functional analyses. PK parameters of LBDE were in good agreement with those of NESP®. CONCLUSION: LBDE shows high similarity to NESP® with regard to structure and function.

The effect of sacroiliac joint mobilization on pelvic deformation and the static balance ability of female university students with si joint dysfunction.

[Purpose] The present study aimed to determine the effect of an 8-week program of joint mobilization on changes in pelvic obliquity and pain level in seventeen female university students aged in their 20's with sacroiliac joint dysfunction by dividing them into two groups: a joint mobilization group (MWM) and a control group. [Subjects] Seventeen subjects were selected from female university students aged in their 20's attending N University in Cheon-An City, Korea, The subjects had sacroiliac joint syndrome, but experienced no problems with daily living and had no previous experience of joint mobilization exercise. The subjects were randomly assigned to a joint mobilization group of eight and a control group of nine who performed joint mobilization exercise. [Methods] Body fat and lean body mass were measured using InBody 7.0 (Biospace, Korea). The Direct Segmental Multi-frequency Bioelectrical Impedance Analysis Method (DSM-BIA) was used for body composition measurement. A pressure footstool (Pedoscan, DIERS, Germany) and a trunk measurement system (Formetric 4D, DIERS, Germany), a 3D image processing apparatus with high resolution for vertebrae, were used to measure 3D trunk images of the vertebrae and pelvis obliquity, as well as static balance ability. [Result] The MWM group showed a significantly better Balance than the control group. In addition, the results of the left/right and the front/rear balance abilities were significantly better than those of the control group. [Conclusion] This study proved that a combination of mobilization with movement and functional training was effective in reducing pelvis malposition and pain, and improving static stability control.

Sterols isolated from seeds of Panax ginseng and their antiinflammatory activities.

BACKGROUND: Panax ginseng C. A. Meyer, a perennial herb from the Araliaceae family, is a commonly used medicinal plant. Many studies have been conducted on the biologically active constituents of whole parts of P. ginseng (i.e., roots, leaves, flower buds, and fruits). However, the seeds of P. ginseng have not been intensively investigated. A new sterol glucoside,3-O-b-d-glucopyranosyl-5,22,24-stigmastatrienol (1), and a known sterol, 5,22-stigmastadienol (2), were isolated from seeds of P. ginseng and were evaluated for their inhibitory activities on tumor necrosis factor (TNF)α-induced nuclear factor (NF)-κB and inducible nitric oxide synthase (iNOS) transcription in transfected HepG2 cells. The present work deals with the isolation, identification, and antiinflammatory activities of the two compounds. MATERIALS AND METHODS: The compounds were isolated by a combination of silica gel and YMC R-18 column chromatography, and their structures were identified by analysis of spectroscopic data (1D, 2D-NMR, and MS). The antiinflammatory activities of the isolated compounds 1 and 2 were evaluated by luciferase reporter gene assays. RESULTS: Two sterols have been isolated from the seeds of P. ginseng. Compound 1 is a previously unreported glucosidyl sterol. Compounds 1 and 2 both inhibited NFκB-luciferase activity, with IC50 values of 8.1 and 4.8΅M, respectively. They also inhibited iNOS-luciferase activity in TNFα-induced HepG2 cells, with IC50 values of 2.2 and 2.9΅M, respectively. CONCLUSION: The two isolatedsterols have inhibitory effects on inflammation-related factors in HepG2 cells, as determined by luciferase reporter gene assays. Thus, seeds of P. ginseng are worthy of consideration for the development and research of antiinflammatory agents.

A new lupane-type triterpene from the seeds of Panax ginseng with its inhibition of NF-κB.

A new lupane-triterpene, 3ß-cis-feruloyloxy-16ß-hydroxylup-20(29)-ene (1) were isolated from the ethyl acetate extract of Panax ginseng seeds along with the known compound, 3ß-transferuloyloxy-16ß-hydroxylup-20(29)-ene (2). Compound 2 was isolated from this plant for the first time. Their chemical structures were determined by mass spectroscopy and one-dimensional and two-dimensional magnetic resonance spectra. The bioactive effects of these compounds on TNF-α-induced NF-κB transcription were evaluated in transfected HepG2 cells. Effects on the expression of NF-κB target genes were also examined using a reverse transcription-polymerase chain reaction. Both compounds 1 and 2 were inhibited NF-κB activity in HepG2 cells by decreasing the cellular concentrations of inflammatory factors iNOS and COX2.

Isoconiferoside, a new phenolic glucoside from seeds of Panax ginseng.

A new phenolic glucoside, isoconiferoside (1), was isolated from the seeds of Panax ginseng (Araliaceae). The structure was determined to be 9-O-[ß-D-glucopyranosyl-(1 --> 6)-ß-D-glucopyranosyl]-trans-coniferyl alcohol based on spectroscopic analyses (1H- and 13C-NMR, DEPT, COSY, HMQC, and HMBC) and acid hydrolysis.

Inhibitory effect of ginsenosides from steamed ginseng-leaves and flowers on the LPS-stimulated IL-12 production in bone marrow-derived dendritic cells.

Interleukin-12, a heterodimeric cytokine comprising p40 and p35 subunits, plays an essential role in the regulating the differentiation of Th cells, which establish and maximize the capabilities of the immune system. The aim of present study is to screen the effect of 21 ginsenosides from steamed ginseng-leaves and flowers on IL-12 production in bone marrow-derived dendritic cells induced by lipopolysaccharide. Noticeably, ginsenoside Rg(6) (12) and ginsenoside F(4) (13) exhibited particularly inhibitory effect on LPS-induced IL-12 production with the inhibition values of 80 and 82%; and ginsenoside ST(1) (4), ginsenoside SL(2) (8), ginsenoside SL(3) (9), ginsenoside Rh(3) (14), ginsenoside Rk(2) (15), and ginsenoside Rs(4) (18) showed moderate effects with inhibition rates of 63, 65, 67, 68, 71, 73, and 67%, respectively. These results warrant further studies concerning potential of saponin extracts of steamed ginseng-leaves and flowers for medicinal uses.