Discovery of Novel Cholinesterase Inhibitors Easily Crossing the Blood-Brain Barrier via Structure-Property Relationship Investigation: Methylenedioxy-Cinnamicamide Containing Tertiary Amine Side Chain.

In the present investigation, a series of dimethoxy or methylenedioxy substituted-cinnamamide derivatives containing tertiary amine moiety (N. N-Dimethyl, N, N-diethyl, Pyrrolidine, Piperidine, Morpholine) were synthesized and evaluated for cholinesterase inhibition and blood-brain barrier (BBB) permeability. Although their chemical structures are similar, their biological activities exhibit diversity. The results showed that all compounds except for those containing morpholine group exhibited moderate to potent acetylcholinesterase inhibition. Preliminary screening of BBB permeability shows that methylenedioxy substituted compounds have better brain permeability than the others. Compound 10c, containing methylenedioxy and pyrrolidine side chain, showed a better acetylcholinesterase inhibition (IC50: 1.52±0.19 µmol/L) and good blood-brain barrier permeability. Further pharmacokinetic investigation of compound 10c using ultra high performance liquid chromatography-mass/mass spectrometry (UPLC-MS/MS) in mice showed that compound 10c in brain tissue reached its peak concentration (857.72±93.56 ng/g) after dosing 30 min. Its half-life in the serum is 331 min (5.52 h), and the CBrain/CSerum at various sampling points is ranged from 1.65 to 4.71(Mean: 2.76) within 24 hours. This investigation provides valuable information on the chemistry and pharmacological diversity of cinnamic acid derivatives and may be beneficial for the discovery of central nervous system drugs.

Corrigendum: Tree-shrub-grass composite woodland better facilitates emotional recovery in college students emotion better than other plant communities.

[This corrects the article DOI: 10.3389/fpsyg.2024.1285792.].

Tree-shrub-grass composite woodland better facilitates emotional recovery in college students emotion better than other plant communities.

Previous research has indicated that natural landscapes exhibit a greater capacity for ameliorating negative emotional states in individuals when compared to urban landscapes. Nevertheless, significant scientific inquiries, such as the uniformity of the rejuvenating effect across distinct categories of natural landscapes on college students and the choice of the optimal plant community for achieving the most potent restorative effect, remain unexplored. This study aimed to address these questions by selecting four plant communities (single-layer grassland, single-layer woodland, tree-grass composite woodland, tree-shrub-grass composite woodland) and using an electroencephalography method to capture the neuroelectric activity of the participants in combination with the Positive and Negative Affect Schedule score to explore the effects of plant community types on emotional recovery. The results showed that all four plant communities significantly increased positive emotions and significantly reduced negative emotions. There was no significant difference in the recovery effect of positive emotions among the four plant community types, but there was a significant difference in the recovery effect of negative emotions. The effect of tree-shrub-grass composite woodland on the negative emotion recovery effect is the best; the EEG results found that the alpha wave amplitude induced by the tree-shrub-grass composite woodland was significantly higher than that of the other three groups of plant communities, and the EEG and behavioral results were consistent. The results show that the tree-shrub-grass composite woodland has the best restoration effect and has stronger planning and design significance.

Design, synthesis and the evaluation of cholinesterase inhibition and blood-brain permeability of daidzein derivatives or analogs.

In the present study, a series of derivatives and analogs of daidzein were designed and synthesized to investigate cholinesterase inhibition and blood-brain barrier permeability. The enzyme assay showed that most of the compounds containing a tertiary amine group exhibit moderate cholinesterase inhibition, 7-hydroxychromone derivatives (absence of B ring of daidzein scaffold) only have a weaker bioactivity, while those compounds without the tertiary amine group have no bioactivity. Among them compound 15a (4'-N,N-dimethylaminoethoxy-7-methoxyisoflavone) appeared the best inhibitory activity (IC50 : 2.14 ± 0.31 µmol/L) and higher selectivity for AChE over BuChE (Ratio: 7.07). It was selected for the further investigation by UPLC-MS/MS. The results show that CBrain/Serum of compound 15a in mice was more than 2.87 within 240 min. This discovery may provide worthy information for the future development of central nervous drugs including but not limited to cholinesterase inhibitors.

Bio-based poly(hexamethylene 2,5-furandicarboxylate-co-2,6-naphthalate) copolyesters: a study of thermal, mechanical, and gas-barrier properties.

A series of poly(hexamethylene 2,5-furandicarboxylate-co-2,6-naphthalate) copolyesters were synthesized using various amounts of poly(hexylene 2,5-furandicarboxylate) (PHF) and poly(hexylene 2,6-naphthalate) (PHN) via melt polymerization. The effects of introducing 2,6-naphthalene dicarboxylic acid (NDCA) on the thermal, mechanical, and gas-barrier properties were investigated. When the NDCA content was less than 30 mol%, the temperatures of crystallization (Tc) and melting (Tm) decreased as the amount of NDCA was increased owing to disturbance of the polymer-chain regularity. When the NDCA content was above 50 mol%, the Tc and Tm of the materials increased as the NDCA content was increased, showing that the dominant crystallization behavior varied from 2,5-furandicarboxylic acid to NDCA. Hence, the glass transition temperature (Tg) increased as the NDCA content was increased, which was attributed to the incorporation of NDCA with a more rigid naphthalate structure compared with the furan ring. The gas-barrier properties of the samples were observed to improve with the introduction of NDCA; this tendency could be explained by the ß-relaxation behavior and free volume values of the samples in the amorphous state. The activation energy (Ea) of ß-relaxation increased with the NDCA content, indicating that higher amounts of energy were needed to trigger the onset of long-range molecular motions. Free-volume calculations of the polymer structure showed that the introduction of NDCA hindered the space for gas penetration. For these reasons, the gas-barrier properties were improved and evaluated.

A Rapid Thermal Absorption Rate and High Latent Heat Enthalpy Phase Change Fiber Derived from Bio-Based Low Melting Point Copolyesters.

A series of poly(butylene adipate-co-hexamethylene adipate) (PBHA) copolymers with different content of 1,4-cyclohexanedimethanol (CHDM) was synthesized via one-step melt polymerization. The PBHA copolymer with 5 mol% CHDM (PBHA-C5) exhibited a low melting point (Tm) and high enthalpy of fusion (∆Hm) of 35.7 °C and 43.9 J g-1, respectively, making it a potential candidate for an ambient temperature adjustment textile phase change material (PCM). Polybutylene terephthalate (PBT) was selected as the matrix and blended at different weight ratios of PBHA-C5, and the blended samples showed comparable Tm and ∆Hm after three cycles of cooling and reheating, indicating good maintenance of their phase changing ability. Samples were then processed via melt spinning with a take-up speed of 200 m min-1 at draw ratios (DR) of 1.0 to 3.0 at 50 °C. The fiber's mechanical strength could be enhanced to 2.35 g den-1 by increasing the DR and lowering the PBHA-C5 content. Infrared thermography showed that a significant difference of more than 5 °C between PBT and other samples was achieved within 1 min of heating, indicating the ability of PBHA-C5 to adjust the temperature. After heating for 30 min, the temperatures of neat PBT, blended samples with 27, 30, and 33 wt% PBHA-C5, and neat PBHA-C5 were 53.8, 50.2, 48.3, 47.2, and 46.5 °C, respectively, and reached an equilibrium state, confirming the temperature adjustment ability of PBHA-C5 and suggesting that it can be utilized in thermoregulating applications.

Low-Mass Liquid Crystalline Materials Blended in Recycled Thermoplastic Polyester Elastomer for Corrosion Inhibitor Application.

In this work, the development and application of multicomponents obtained from recycled polyethylene terephthalate (r-PET) waste and monotropic liquid crystals as anticorrosion coatings are reported. The r-PET raw material was alcoholyzed and reproduced as a thermoplastic polyester elastomer (TPEE) with different amounts (n%, n = 0, 1, 3, and 5) of 1,6-hexanediamine (HDA). Then, a fluorine-containing liquid crystal (4-cyano-3-fluorophenyl 4-ethylbenzoate (4CFE)) was incorporated into the TPEE mixture via solvent blending to modify and enhance the water resistance. The adhesion behavior of the coating on glass and iron substrates was evaluated by cross-cut tests and immersion tests in aqueous NaCl. In the corrosion resistance measurements, all of the coating samples fabricated with 10 ± 1 mm thickness were less active toward electrochemical corrosion (PEF% > 99%) than the bare iron plate, indicating that our work provided better protection against corrosion of the iron plate.

Handgrip strength, tumor necrosis factor-α, interlukin-6, and visfatin levels in oldest elderly patients with cognitive impairment.

INTRODUCTION: Handgrip strength is associated with mild cognitive impairment. Tumor necrosis factor [TNF]-α and interleukin [IL]-6 were pro-inflammatory cytokines influencing the severity of initial neurological deficit. Visfatin is a novel adipokine and has a strong correlation with inflammation. The relationships of TNF-α, IL-6 and visfatin are not consistent, and no study has investigated them in the elderly patients with cognitive impairment. METHODS: This study included patients aged ≥75 years at the emergency department from August 2018 to February 2019. All patients underwent comprehensive geriatric assessment and blood tests for fasting plasma TNF-α, IL-6 and visfatin levels. RESULTS: We enrolled 106 elderly patients with a mean age of 87.3 years, including 62 (58.4%) patients in cognitive impairment group (Mini-Mental State Examination [MMSE] < 24) and 44 (41.5%) patients in the non-cognitive impairment group. Compared to the non-cognitive impairment group, the cognitive impairment group had significantly lower handgrip strength, and significantly higher TNF-α, IL-6 and visfatin levels. TNF-α positively correlated with IL-6. Both TNF-α and IL-6 negatively correlated with Barthel index and MMSE. Handgrip strength negatively correlated with TNF-α but positively correlated with Barthel index and MMSE scores. Backward and stepwise multiple logistic regression analyses showed that the independent predictor for cognitive impairment was handgrip strength and age. CONCLUSION: The cognitive impairment group had significantly higher serum TNF-α, IL-6, and visfatin levels. The independent predictors of cognitive impairment were handgrip strength and age. Handgrip strength negatively correlated with TNF-α and IL-6 but positively with Barthel index and MMSE scores.

Synthesis of Bio-Based Poly(Butylene Adipate-co-Butylene Itaconate) Copolyesters with Pentaerythritol: A Thermal, Mechanical, Rheological, and Molecular Dynamics Simulation Study.

Bio-based unsaturated poly(butylene adipate-co-butylene itaconate) (PBABI) aliphatic copolyesters were synthesized with pentaerythritol (PE) as a modifier, observing the melting point, crystallization, and glass transition temperatures were decreased from 59.5 to 19.5 °C and 28.2 to -9.1 °C as an increase of itaconate concentration, and Tg ranged from -54.6 to -48.1 °C. PBABI copolyesters tend to the amorphous state by the existence of the BI unit above 40 mol%. The yield strength, elongation, and Young's modulus at different BA/BI ratios were valued in a range of 13.2-13.8 MPa, 575.2-838.5%, and 65.1-83.8 MPa, respectively. Shear-thinning behavior was obtained in all BA/BI ratios of PBABI copolyesters around an angular frequency range of 20-30 rad s-1. Furthermore, the thermal and mechanical properties of PBABI copolyesters can be well regulated via controlling the itaconic acid contents and adding the modifier. PBABI copolyesters can be coated on a 3D air mesh polyester fabric to reinforce the mechanical property for replacing traditional plaster applications.

Influence of asymmetric substituent group 2-methyl-1,3-propanediol on bio-based poly(propylene furandicarboxylate) copolyesters.

A series of bio-based poly(propylene-co-2-methyl-1,3-propanediol 2,5-furandicarboxylate) (PPMF) copolyesters, with various compositions from poly(propylene 2,5-furandicarboxylate) (PPF) to poly(2-methyl-1,3-propylene 2,5-furandicarboxylate) (PMePF), were synthesized by conventional melt polymerization. The effects of the substituent group to PPF on the thermal properties, mechanical properties, and gas barrier properties were analyzed by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic mechanical analysis (DMA), tensile testing, and the oxygen permeation test. The introduction of the methyl group with a hydrogen atom altered the thermal behavior and gas barrier properties of copolyesters, suggesting that the glass temperature (Tg) and the melting temperature (Tm) were decreased as the 2-methyl-1,3-propanediol (MPO) content increased. PPF exhibited the highest Tm and Tg of 175.9 °C and 83.0 °C with a melting enthalpy (ΔHm) of 38.6 J g-1, and poly(2-methyl-1,3-propylene 2,5-furandicarboxylate) formed as an amorphous polyester. Moreover, the effect of a substituent methyl group on the barrier properties was attributed to ß relaxation and fraction free volume, which could be raised by replacing the methyl group with a hydrogen atom for PPF, weakening the barrier properties.

A modified multiscale peak alignment method combined with trilinear decomposition to study the volatile/heat-labile components in Ligusticum chuanxiong Hort - Cyperus rotundus rhizomes by HS-SPME-GC/MS.

Head Space/Solid Phase Micro-Extraction (HS-SPME) coupled with Gas Chromatography/Mass Spectrometer (GC/MS) was used to determine the volatile/heat-labile components in Ligusticum chuanxiong Hort - Cyperus rotundus rhizomes. Facing co-eluting peaks in k samples, a trilinear structure was reconstructed to obtain the second-order advantage. The retention time (RT) shift with multi-channel detection signals for different samples has been vital in maintaining the trilinear structure, thus a modified multiscale peak alignment (mMSPA) method was proposed in this paper. The peak position and peak width of representative ion profile were firstly detected by mMSPA using Continuous Wavelet Transform with Haar wavelet as the mother wavelet (Haar CWT). Then, the raw shift was confirmed by Fast Fourier Transform (FFT) cross correlation calculation. To obtain the optimal shift, Haar CWT was again used to detect the subtle deviations and be amalgamated in calculation. Here, to ensure there is no peaks shape alternation, the alignment was performed in local domains of data matrices, and all data points in the peak zone were moved via linear interpolation in non-peak parts. Finally, chemical components of interest in Ligusticum chuanxiong Hort - Cyperus rotundus rhizomes were analyzed by HS-SPME-GCMS and mMSPA-alternating trilinear decomposition (ATLD) resolution. As a result, the concentration variation between herbs and their pharmaceutical products can provide a scientific basic for the quality standard establishment of traditional Chinese medicines.

Accurate recognition and feature qualify for flavonoid extracts from Liang-wai Gan Cao by liquid chromatography-high resolution-mass spectrometry and computational MS/MS fragmentation.

In this study, Liquid Chromatography (LC) separation combined with quadrupole-Time-Of-Flight Mass Spectrometry (qTOF-MS) detection was used to analyze the characteristic ions of the flavonoids from Liang-wai Gan Cao (Radix Glycyrrhizae uralensis). First, accurate mass measurement and isotope curve optimization could provide reliable molecular prediction after noise deduction, baseline calibration and "ghost peak recognition". Thus, some spectral features in the LC-MS data could be clearly explained. Secondly, the chemical structure of flavonoids was deduced by MS/MS fragment ions, and the in-silico spectra by MS-FINDER program provided strong support for overcoming the bottleneck of phytochemical identification. For a predicted formula and experimental MS/MS spectrum, the MS-FINDER program could sort the candidate compounds in the public database based on a comprehensive weighted score, and we took the first 20 reliable compounds to seek the target compound in an in-house database. Certainly, those fragmentation pathways could also be deduced and described as Retro-Diels-Alder (RDA) fragmentation reaction, losses of C4H8, C5H8, CH3, CO, CO2 and others. Accordingly, 63 flavonoids were identified, and their in-silico bioactivity were clearly disclosed by some bioinformatics tools. In this experiment, the flavonoids obtained by the four extraction processes were tested by LC-qTOF-MS. We looked for possible Q-markers from these data matrices and then quantified them; their similarities/differences were also described. The results also indicated that the Macroporous Adsorption Resins (MARs) purification is a low cost, environmentally friendly and effective approach.

Chemometrics-enhanced one-dimensional/comprehensive two-dimensional gas chromatographic analysis for bioactive terpenoids and phthalides in Chaihu Shugan San essential oils.

Chemometrics-enhanced one-dimensional/comprehensive two-dimensional gas chromatographic (GC/GC×GC) technologies, were used to explore the compositions of Chaihu Shugan San essential oils, that were extracted from the herbal formulae by different schemes. We have shown that chemometric resolution using gas chromatographic- mass spectrometry (GC-MS) could be used for the qualitative and quantitative analysis of the majority of Terpenoids or Phthalides from herb formulae and single herbs. A GC×GC system was further optimized to achieve the increased peak capacity and the enhanced signal of the hydro-distillation sample (CSSh). When hardware bottleneck resulted from very complex sample, chemometric tools were once again applied to recover the stained information in the second dimension (2D) matrix data. Heuristic evolving latent projections (HELP) could be used for two dimensional (2D) sub-matrixes Xi at n spectral detection channels, after three dimensional (3D) data splitting. For a real 3D data matrix, alternating trilinear decomposition (ATLD) algorithm could conduct regularization for an iterative trilinear decomposition procedure, by Moore-Penrose pseudoinverse computations based on singular value decomposition. After retention indices (RI) confirmation, 216 target analytes (terpenoids or phthalides) could be elucidated both in CSSh and in supercritical fluid extract (CSSs). Based on the obtained data, some potential quality markers (Q-markers) were identified which may affect the quality of the products. Finally, a "connectivity map" was plotted to describe the unique mechanisms of tradition Chinese medicine (TCM).

Obeticholic acid improves hepatic steatosis and inflammation by inhibiting NLRP3 inflammasome activation.

BACKGROUND AND AIM: Several pre-clinical and clinical researches have proved that obeticholic acid (OCA)has a potential therapeutic effect on non-alcoholic steatohepatitis (NASH). Our aim was to investigate whether the therapeutic effect of OCA on NASH was attributed to its inhibition effect on cytosolic sensor NLR family pyrin domain containing 3 (NLRP3) inflammasome activation. METHODS: We used mice model of methionine-choline-deficient (MCD) diet induced NASH. At different fibrosis stages, the expressions of NLRP3, caspase-1 and IL-1ß were analyzed by means of immunohistochemistry and western blot respectively. After daily gavage of 0.4 mg of OCA or vehicle for 24 days, we evaluated the direct effect of OCA on NLRP3 inflammasome activation by analyzing the expressions of NLRP3 and IL-1ß. Additionally, liver function and liver histology of mice were assessed. The expressions of NLRP3 and IL-1ß above and the expressions of fibrosis-related genes were analyzed by quantitative real-time polymerase chain reaction (PCR). RESULTS: NLRP3 inflammasome activation could be observed in liver fibrosis, and we found that the expressions of NLRP3, caspase-1 and IL-1ß gradually increased to peak at stage 2-3 but decreased significantly at stage 4 of liver fibrosis in MCD mice model. We also found that short-term OCA treatment could significantly down-regulate the expressions of NLRP3 and IL-1ß and therefore improved NASH-associated steatosis and inflammation. CONCLUSIONS: NLRP3 inflammasome could be activated and might have an essential role in NASH progression, and short-term OCA treatment could have a potential therapeutic effect on NASH-associated steatosis and inflammation by inhibiting NLRP3 inflammasome activation.

GC-MS Analysis and Volatile Profile Comparison for the Characteristic Smell from Liang-wai Gan Cao (Glycyrrhiza uralensis) and Honey-Roasting Products.

The characteristic smell of Liang-wai Gan Cao (Glycyrrhiza uralensis) and honey-roasting products was comprehensively analyzed using gas chromatography-mass spectrometry (GC-MS). Steam distillation and headspace solid-phase microextraction (HS-SPME) were used to extract volatile organic compounds (VOCs). Multiple fibers of SPME may reflect the samples' comprehensive information to the greatest extent, depending on their chemical characters. After chemometric resolution and spectra interpretation, many aroma compounds could be identified from GC-MS data. As a result, principal component analysis was set for the differentiation of several G. uralensis samples in different regions, and some important peaks could be found. Next, VOCs' profiles of honey-roasting products suggested that the flavors could be influenced by honey and pharmaceutical technologies.

Development and characterization of a novel fusion protein of a mutated granulocyte colony-stimulating factor and human serum albumin in Pichia pastoris.

The purpose of the present work was to develop a novel, long-acting and potent human serum albumin/granulocyte colony stimulating factor (HSA/G-CSF) therapeutic fusion protein. The novel fusion protein, called HMG, was constructed by genetically fusing mutated human derived G-CSF (mG-CSF) to the C-terminal of HSA and then prepared in Pichia pastoris. The molecular mass of HMG was about 85 kDa and the isoelectric point was 5.3. Circular dichroism spectroscopy suggested that mG-CSF retained nearly all of its native secondary structure, regardless of fusion. The binding capabilities of mG-CSF moiety to G-CSF receptor and HSA moiety to warfarin showed very little change after fusing. The bioactivity of HMG (11.0×10(6) IU/mg) was more than twice that of rHSA/G-CSF (4.6×10(6) IU/mg). A mutation was made at the 718th amino acid of HMG, substituting Ala for Thr, to investigate the glycosylation of HMG expressed in P. pastoris. Data indicated that HMG was modified at Thr718, speculatively with the addition of a mannose chain. In conclusion, a novel HSA/G-CSF fusion protein was successfully constructed based on a mutated G-CSF. This protein showed more potent bioactivity than rHSA/G-CSF and thus may be a suitable long-acting G-CSF.

Controllable growth of CNTs on graphene as high-performance electrode material for supercapacitors.

Design and synthesis of three-dimensional (3D) structured carbon materials are crucial for achieving high-performance supercapacitors (SC) for energy storage. Here, we report the preparation of 3D architectured GN-CNT hybrid as SC electrodes. Controllable growth of carbon nanotubes on graphene sheets was realized through a facile one-pot pyrolysis strategy. The length of the carbon nanotubes could be rationally tuned by adjusting the amount of precursors. Correspondingly, the resulted GN-CNT hybrid showed adjustable electrochemical performance as an SC electrode. Importantly, the GN-CNT exhibited a high specific surface area of 903 m(2) g(-1) and maximum specific capacitance of 413 F g(-1) as SC electrodes at a scan rate of 5 mV s(-1) in 6 M KOH aqueous solution. This work paves a feasible pathway to prepare carbon electrode materials with favorable 3D architecture and high performance, for use in energy storage and conversion.

Engineering a pharmacologically superior form of granulocyte-colony-stimulating factor by fusion with gelatin-like-protein polymer.

The plasma half-life of therapeutic proteins is a critical factor in many clinical applications. Therefore, new strategies to prolong plasma half-life of long-acting peptides and protein drugs are in high demand. Here, we designed an artificial gelatin-like protein (GLK) and fused this hydrophilic GLK polymer to granulocyte-colony-stimulating factor (G-CSF) to generate a chimeric GLK/G-CSF fusion protein. The genetically engineered recombinant GLK/G-CSF (rGLK/G-CSF) fusion protein was purified from Pichia pastoris. In vitro studies demonstrated that rGLK/G-CSF possessed an enlarged hydrodynamic radius, improved thermal stability and retained full bioactivity compared to unfused G-CSF. Following a single subcutaneous administration to rats, the rGLK/G-CSF fusion protein displayed a slower plasma clearance rate and stimulated greater and longer lasting increases in circulating white blood cells than G-CSF. Our findings indicate that fusion with this artificial, hydrophilic, GLK polymer provides many advantages in the construction of a potent hematopoietic factor with extended plasma half-life. This approach could be easily applied to other therapeutic proteins and have important clinical applications.

Preparation and characterization of a potent, long-lasting recombinant human serum albumin-interferon-alpha2b fusion protein expressed in Pichia pastoris.

A long-lasting recombinant human serum albumin-interferon-alpha2b fusion protein (rHSA/IFNalpha2b) was prepared and its structure and biological activities were studied. rHSA/IFNalpha2b was expressed in methylotrophic yeast Pichia pastoris with HSA's natural signal peptide and purified by dye affinity chromatography, hydrophobic interaction chromatography, ion exchange chromatography and Sephadex G25. Purity of the prepared rHSA/IFNalpha2b was greater than 97% analyzed by non-reduced SDS-PAGE and RP-HPLC. Structure and biological activities of the prepared rHSA/IFNalpha2b were characterized by physical, chemical and biological methods. Its pI was 5.3 and showed a single band on IEF gel. Molecular weight determined by MALDI-TOF was 86004.3+/-29.2. Amino-terminal and carboxyl-terminal amino acid sequences were identical to predicted sequence. Its specific activity in vitro was 6.3+/-0.8x10(5) IU/mg fusion protein, retaining about 1.4% of that of unmodified rIFNalpha on a molar basis. After administered in monkeys, significant increases of 2',5'-oligoadenylate synthetase activity relative to IFN-alpha were maintained for 14 days in serum and the rHSA/IFNalpha2b showed more potent biological activity than IFN-alpha on a molar basis. Therefore, markedly improved in vivo biological activity of rHSA/IFNalpha2b could exhibit more potent antiviral activity than IFNalpha2b in future clinical trials.

[Expression in Pichia pastoris and properties of human serum albumin-interferon alpha2b chimera].

To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.