Over the past four decades, monoclonal antibodies (MAbs) have evolved from bioscience research tools to powerful biopharmaceutical MAbs products for multiple diseases treatment. More than 50 therapeutic MAbs have been approved by FDA, widely used in cancer, autoimmune diseases and other diseases in current market. This article reviews the current progress of MAbs development technology, key molecules for cancer-targeted therapy and immunotherapy, and emphasizes the importance of MAbs for disease diagnosis and treatment.

The Hedgehog signaling pathway plays a key role in mammalian embryogenesis, while it is quiescent in adult tissues. The aberrant activation of Hedgehog signaling pathway has been linked to multiple types of malignant tumors, which makes it an attractive target for cancer therapy in recent years. Up to now, two Hedgehog inhibitors (vismodegib and sonidegib) have been proved by FDA for the treatment of tumors. However, drug resistance and severe adverse reactions represent the problems in their clinical application. Novel inhibitors targeting Smo, such as taladegib, NVP-LEQ506, MRT-92, and downstream or upstream of Smo receptor such as Shh ligand and Gli have been developed to overcome the drug resistance and adverse reactions. The current Hedgehog inhibitors are used for treatment of basal cell carcinoma only, while many ongoing clinical trials are conducted to investigate the antitumor effect of Hedgehog inhibitors in other malignancies. Here we reviewed the research progress of the new anticancer drugs targeting the Hedgehog signaling pathway and their prospect in variety of cancers therapy.

Cryptococcal meningitis, caused by fungus Cryptococcus neoformans, is responsible for over a million infections and 600 000 deaths annually. Largely due to the limited treatment options and the intrinsic drawbacks coupled with drug resistance to current therapies, it is urgent to discover novel antifungal agents against cryptococcosis. An ideal antifungal drug should at least satisfy the following criteria: fungicidal, fungus-specific, permeable for the host barriers such as cell membranes of phagocytes and the blood-CNS barrier. Both discovery of candidates with novel mode of action and repurposing existing molecules with potent anti-cryptococcal activity are effective ways in discovery of new anti-cryptococcal agents. Here, we summarized recent advances in the study of anti-fungal activities, mechanisms of action, and clinical developments of new anti-cryptococcal drugs.

Drug transporters play vital roles in absorption, distribution and excretion of drugs. Understanding the transport activity can improve the effectiveness and safety of drugs and guide clinical rational use of drugs. Metformin is a first-line drug in the treatment of type 2 diabetes mellitus, of which the pharmacokinetics involves several transporters. The changes in expression and function of these transporters affect directly the pharmacokinetics/pharmacodynamics of metformin. This paper reviews the research progress of pharmacokinetics of metformin based on transporters, and these transporters are organic cation transporters (OCTs), multidrug and toxin extrusion proteins (MATE), plasma membrane monoamine transporter protein (PMAT), serotonin reuptake transporter (SERT), thiamine transporter 2 (THTR-2), and carnitine/organic cation 1 (OCTN1).

The radiation is regarded as the fourth biggest pollution following the water, air and noise pollution, which generates a broad impact on human physiology and healthy. The radiation mainly comes from medical rays, industrial rays, nuclear wastes and atmospheric ultraviolet rays. The universality makes people pay more and more attention to the damage effect and mechanism of radiation. The radiation is divided into ionizing and non-ionizing radiation. It is a good idea to study the effect of ionizing and non-ionizing radiation on drug's metabolism, which may give a guidance to clinical medication to avoid adverse reactions and to support personalized medicine. This article reviews the effect of ionizing radiation (γ-rays, X-rays, uranium and cesium) and non-ionizing radiation (ultraviolet rays) on drug metabolism, their impact on metabolic characteristics of some drugs and the impact on expression of enzymes and transporters in drug metabolism, which is conducted with a focus on clinical significance.

Mitochondrion is an important organelle in cells and plays a crucial role in tumor development. Therefore, improvement of the targeting ability of anticancer drugs to mitochondria will increase the treatment efficacy for tumor and reduce the side effect on normal tissues. Here, research progresses in mitochondrial targeting have been reviewed in three aspects for tumors treatment: the potential, permeability and translocase of mitochondrial membrane. The review provides a reference for the development of mitochondria targeted therapeutic systems.

Although multiple studies have shown that matrine can inhibit the proliferation of hepatoma cells, its mechanism of action has not been systematically investigated. In this study, the effects of matrine on the proliferation and migration of human hepatoma SMMC-7721 cells were investigated. Based on this result, anti-hepatoma target-functionally related protein interaction network of matrine was constructed, and topological analysis and clustering analysis were performed to predict the crucial targets of matrine for the anti-hepatoma effects. Pathway enrichment analysis was performed on the validated targets to predict the crucial pathways of matrine. Parts of the crucial proteins were examined by Western blot. Cellular experiments showed that matrine at concentrations of 1, 2 and 4 mg·mL^-1 significantly inhibited the proliferation of SMMC-7721 cells, and matrine at concentrations of 0.5, 1 and 2 mg·mL^-1 significantly inhibited the migration of SMMC-7721 cells. The results of network pharmacology suggest that matrine exerts its anti-hepatoma effects through acting on the key validated targets of heparanase (HPSE), caspase 3 (CASP3), Myc proto-oncogene protein (MYC), matrix metalloproteinases 2 (MMP2) and predicted targets of carbonic anhydrase 1 (CA1), lithostathine 1 alpha precursor (REG1A), carboxylesterases 1 (CES1) and acetaldehyde dehydrogenase 2 (ALDH2), and invasion and migration associated pathways. Western blot results suggest that matrine can down-regulate the expression of MMP2 and up-regulate the expression of CASP3. In this paper, we applied network pharmacology to explain the targets and pathways of matrine against hepatoma. The results provide a scientific basis for elucidation of the mechanisms of matrine against hepatoma.

The purpose of this study was to develop a screening method to determine the activity and selectivity of SGLT2 inhibitor. Human SGLT1/SGLT2 cDNA was inserted into the pMSCVpuro mammalian expression vector and the plasmid was transfected into HEK293 cells. Stably transfected clones were selected in puromycin containing medium. To evaluate the expression of human SGLT1 and SGLT2 in stable transfected cells, RT-PCR, Western blot and immunofluorescence analysis were performed. 1-[N-(7-Nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]-1-deoxy-D-glucose (1-NBDG) was used as a substrate in the uptake assay to evaluate the Na⁺ dependent glucose transport activities of SGLT1/2. The inhibitory activity and selectivity of dapagliflozin/phloridzin were also determined, respectively. The hypoglycemic efficacy of dapagliflozin was evaluated in mice with normal blood glucose and mice with alloxan-induced T1DM. The result showed that SGLT1 was overexpressed in pMSCVpuro-SGLT1 transfected HEK293 cells. SGLT2 protein was overexpressed in pMSCVpuro-SGLT2 transfected HEK293 cells and located in both cytoplasm and membrane. The Na⁺ dependent 1-NBDG uptake was significantly increased in pMSCVpuro-SGLT1/SGLT2 transfected cells compared to that in pMSCVpuro-null transfected cells. The selectivity of dapagliflozin, whose half maximal inhibitory concentration (IC₅₀) for SGLT2 (2.24×10^-10 mol·L^-1) was far lower than that for SGLT1 (6.20×10^-7 mol·L^-1), was better than that of phloridzin. The oral glucose tolerance was elevated after a single dose of dapagliflozin in normal mice. In T1DM mice, compared with model group, no-fasting glucose level was decreased at 1 h after administration and maintained at a lower level for 24 h in a dose-dependent manner. A 20-day administration with dapagliflozin dose-dependently improved the hyperglycemia status. Taken together, a system to evaluate the activity and selectivity of SGLT2 inhibitors was established using 1-NBDG in vitro and the hypoglycemic efficacy in vivo in this study. The advantages of this system include non-radioactivity, high efficiency, and good stability which may provide a technique platform for development of novel SGLT2 inhibitors.

Annexin is a protein of evolutionarily conserved polygene family that binds to cell membrane phosphatidylserine (PS). PS is closely related to many diseases with a potential as a new drug target. Annexin has a good value in drug discovery and new drug development. Annexin A4 is a member of the annexins family. Annexin A4 involves in a number of cellular functions, such as exocytosis and coagulation. These functions are related to binding of annexin to acidic phospholipids. However, the detail function(s) of annexin A4 has not been fully uncovered. Production of annexin A4 in large quantity is prerequisite for indepth investigation of the structure-function relationship of annexin A4. Human annexin A4 was originally purified from the natural resource at a low yield due to the complex procedure. In the present study, annexin A4 was expressed in a prokaryotic system with a high yield of soluble protein. The plasmid pET28a-annexin A4-EGFP was constructed for the expression. Recombinant annexin A4-EGFP was purified using two methods. Affinity chromatography approach gave a protein yield at purity of 80%. While, the membrane absorption method produced the protein with the purity over 90%. Flow cytometric analysis showed that the annexin A4-EGFP fusion protein could recognize and bind to the apoptotic cells with an affinity PS at 79.58±11.68 nmol·L^-1, which is at the same order of magnitude as A5-EGFP. We successfully achieved the efficient expression of annexin A4-EGFP in prokaryotic system, and provided an easy and convenient method for purifying a large amount of annexin A4-EGFP with a high purity. This study has laid a solid foundation for our study of the function of annexin A4 in the future.

L-Proline-m-bis (2-chloroethyl) amino-L-phenylalanyl-L-norvaline ethyl ester hydrochloride (MF13) is a new anticancer tripeptide. Our previous study in vitro and in vivo showed that MF13 had anti-proliferative activities in a panel of human hepatocellular carcinoma (HCC) cell lines from different origin. In the present study, we focused on the inhibition effect on HCC of MF13 combined with other anti-cancer drugs. The results of combination chemotherapy in vitro indicated that the combination of MF13 with mitomycin C (MMC) at appropriate concentrations led to a synergistic effect; however, the combination of MF13 with vincristine (VCR) showed no synergistic effect. In the Bel-7402 tumor bearing nude mice, the antitumor effect of the groups of 2 mg·kg^-1 MF13 + 2 mg·kg^-1 MMC or 2 mg·kg^-1 MF13 + 50 mg·kg^-1 cyclophosphamide (CTX) exhibited synergistic anticancer efficacies while the group of 2 mg·kg^-1 MF13 + 0.3 mg·kg^-1 VCR did not have the same effect. Based on our data, we believe that MF13 can be considered as a potential agent against human hepatocellular carcinoma no matter how treated, alone or combined with other drugs.

IG-105, N-(2, 6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide, a novel antimicrotubule agent, showed potent anticancer activity in a variety of human tumor cells in vitro and in vivo. In order to characterize the metabolism and the possible drug-drug interaction of IG-105, we carried out a series of experiments. Drug metabolizing enzymes involved in IG-105 metabolism were investigated by using pooled human liver microsomes (HLMs) and recombinant cytochrome P450 isoforms (rP450s) respectively. The possible metabolites were analyzed by liquid chromatography-orbitrap-mass spectrometry (LC-Orbitrap-MS). The inhibitory effect of IG-105 on main P450 enzymes was also evaluated. The results showed that IG-105 can be metabolized by a series of rP450s, including CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4 and CYP3A5, with the major contribution enzymes being CYP1A2, CYP2B6, CYP2C19, and CYP3A. Three metabolites (M1-M3) were identified and demethylation was the major phase I metabolic reaction for IG-105. IG-105 moderately inhibited CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A enzyme activities with IC₅₀ values of 6.42, 23.64, 0.39, 1.4, and 3.14 μmol·L^-1, respectively. Since the biotransformation of IG-105 involves multiple enzymatic pathways, the compound is less likely to be a victim of a concomitantly used medicine which inhibits activity of one of the CYPs. However, as IG-105 showed medium to strong inhibition on CYP1A2, CYP2D6, CYP3A, and CYP2C19, caution is particularly needed when IG-105 is co-administrated with other anticancer drugs which are mainly metabolized by the above enzymes.

Studies have shown that acetylcholinesterase inhibitors donepezil and galantamine have effects of reducing neuronal damage caused by glucose deprivation and reducing the cerebral infarction volume of cerebral ischemic animals, but their effects may not be entirely dependent on its inhibition of cholinesterase activity. In order to study the effects of donepezil and galantamine on neuronal injury of cerebral ischemia, the rat neuron-astrocyte co-culture model was successfully established in this study. In this model, we studied the effects of donepezil and galantamine on neuron apoptosis induced by oxygen-glucose deprivation/reoxygenation (OGD/R) and investigated the mechanism. The results showed that donepezil and galantamine significantly reduced the neuron apoptosis, and promoted the synthesis and secretion of BDNF and NGF in astrocytes in the co-culture system. Donepezil and galantamine activated the PI3K/Akt pathway and ERK pathway, and promoted the phosphorylation of the nuclear transcription factor CREB. These results suggest that donepezil and galantamine exhibit protective effects on neuronal damage induced by OGD/R. The mechanism may be related to activation of PI3K/Akt pathway and ERK pathway in astrocytes and promote phosphorylation of CREB, which lead to the synthesis and secretion of BDNF and NGF from astrocytes.

Cathepsin K (CTSK) is considered a critical pharmaceutical target in the treatment of osteoporosis. CTSK exerts proteolytic activities against regulatory proteins besides its collagenase function, which may account for some of the adverse reactions when blocked by active site-directed inhibitors. Exosite inhibitors that can discriminate between the therapeutic collagenase and other biological activities of CTSK specifically inhibit the collagenase activity of CTSK without interfering with the other proteolytic activities of the protease. Active recombinant CTSK was expressed in Pichia pastoris, and purified by n-butyl sepharose and SP sepharose column chromatography. Herba Ecliptae is a common traditional Chinese medicine in the treatment of bone diseases. Collagenase assay and benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) substrate assay based on CTSK are applied to verify the exosite inhibitors. n-Butanol extract of Herba Ecliptae are the most active fraction and eclalbasaponin Ⅸ isolated from n-butanol fraction is the potential exosite inhibitor of CTSK.

This study was designed to investigate the chemical constituents of the anti-osteoporotic part of Lepidium meyenii Walp. (maca) produced in Heqing, Yunnan. Seven compounds were isolated from the n-BuOH extract of maca using combination of column chromatographies on MCI resin, silica gel, C₁₈ bonded silica gel, and Sephadex LH-20, followed by semi-preparative HPLC and recrystallization. The purified compounds were identified on the basis of their physicochemical properties and spectral data as macaolidine (1), tryptophan (2), daucosterol (3), (3S)-1, 2, 3, 4-tetrahydro-β-carboline-3-carboxylicacid (4), chlorogenic acid (5), luteolin (6), and hyperoside (7). Compound 1 is a new phenylacetamide alkaloid, and compounds 4-7 were isolated from Lepidium meyenii for the first time.

Two flavanone glucosides were isolated from the 80% ethanol extract of Glycyrrhiza uralensis using various chromatographic techniques, including macroporous adsorbent resin, RP-C₁₈, Sephadex LH-20, MCI and preparative HPLC. On the basis of chemical properties and spectra data, these compounds were identified as (2S)-liquiritigenin-4´-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranoside (1), (2R)-liquiritigenin-4´-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranoside (2), respectively. Compounds 1 and 2 are new compounds, and their aglycones are enantiomers.

Xanthine oxidase (XO) is a key enzyme in the synthesis of uric acid. Therefore, XO inhibitors play an important role in the antihyperuricemic therapy. Based on the template structures of febuxostat and topiroxostat, 18 amide derivatives were designed and synthesized. Among them, six showed apparent inhibitory activity against XO under the concentration of 10 μmol·L^-1. Molecular docking revealed the possible interaction mode of this compound class, which may provide a clue for further molecular design.

This study was designed to establish a high performance liquid chromatography (diode array detector, DAD)-tandem mass chromatography (HPLC-DAD-MS/MS) method for the simultaneously screening 12 kinds of cough-relieving chemical drugs illegally added in anti-cough and antiasthmatic traditional Chinese medicines. This method involved liquid chromatography-tandem mass spectrometry. The separation was conducted by Kromasil100-5C18 (250 mm × 4.6 mm, 5 μm) and the mobile phase was consisted of 0.3% ammonium formate (pH 2.97) and methanol. After separated by HPLC, the suspected components were analyzed by MS/MS and DAD and ultra scan was used to identify these illegally added drugs. A fast and sensitive HPLC-DAD-MS/MS method for the simultaneously screening for illegal chemical compositions was established. The LOD of these substances were below 50 ng. The method was sufficiently selective and sensitive to detect illegal chemical compositions in anti-cough and antiasthmatic traditional Chinese medicines.

This study was designed to establish an ultra-liquid chromatography-mass spectrometry method for the reliable identification the multiple chemical components in Huangqi Jianzhong Tang (HQJZ). The ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry method was applied to identify the chemical constituents in the HQJZ rapidly. A total of 71 compounds including two major categories of saponins and flavonoids were identified or tentatively deduced on the basis of their retention behaviors, fragments of multistage mass spectrometry or by comparing with reference substances and literatures. Among them, 20 compounds were from Astragali Radix, 14 were from Paeoniae Radix Alba, 37 were from Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, 3 were from Rhizoma Zingiberis Recens, 2 were from Cinnamomi Ramulus and Jujubae Fructus, respectively. The LC-MS method was used to qualitatively analyze the chemical constituents of HQJZ, which provides a scientific basis for the quality control of HQJZ.

A method of ultra flow liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was developed to elucidate the impurity of linezolid tablets. Linezolid was subjected to forced degradation under hydrolytic (acid, base and neutral), oxidative, photolytic and thermal. The structure identification of the degradation products and the fragmentation patterns for the related impurities were analyzed. A total of four degradation impurities were characterized, impurity 1 is (S)-1-amino-3-((3-fluoro-4-morpholinophenyl)amino)propan-2-ol, impurity 2 is (S)-4-(4-(5-(acetamidomethyl)-2-oxo-oxazolidin-3-yl)-2-fluorophenyl)morpholine 4-oxide, impurity 3 is (S)-5-(aminomethyl)-3-(3-fluoro-4-morpholinophenyl)oxazolidin-2-one, impurity 4 is (R)-N-(3-((3-fluoro-4-morpholinophenyl)amino)-2-hydroxypropyl)acetamide. Acid degradation induced impurity 3 and impurity 4, base degradation induced impurity 1 and impurity 4, oxidation degradation induced impurity 2, hydrolysis degradation induced impurity 4. The study also determined calibration factor using impurity references, and the calibration factors were found to be 1.3, 1.4, 0.9 and 1.1, respectively. The toxicity of the degradation impurities was predicted by web-based prediction system. The results from this study provide an important reference in quality control and evaluation of linezolid.

This study aims to prepare lipid bilayer-coated calcium phosphate core-shell nanoparticles (LCAPNs), which can dissolve in an acidic environment to improve the tumor cell toxicity of antitumor drug. Paclitaxel (PTX) loaded lipid coated calcium phosphate nanoparticles (PTX-LCAPNs) were prepared by thin-film dispersion method. The morphology, particle size and in vitro release behavior were characterized. Meanwhile, the intracellular uptake, intracellular dissolution, cell toxicity of PTX-LCAPNs and intracellular accumulation of PTX were evaluated in human HCC cell line (Huh-7). The results suggested that the mean diameter of the spherical LCAPNs was 124.73±6.41 nm. The PTX-LCAPNs demonstrated little drug leakage in simulated normal physiological conditions, while a rapid release was observed in simulated intracellular condition in vitro. Moreover, the PTX-LCAPNs achieved 1.7 fold improvement in the intracellular PTX concentration leading to 5-fold reduction in half maximal inhibitory concentration (IC₅₀) values of PTX compared with calcium phosphate nanoparticles loaded with PTX (PTX-CAPNs), demonstrating a stronger cancer cell lethality.

The mesoporous silica nanoparticles (MSN) in different pore size and sirolimus (SRL) loaded self-microemulsifying drug delivery system (SMEDDS) were prepared. The results in morphology were collected by scanning electron microscope, transmission electron microscope, small-angle X-ray diffraction, and N₂ adsorption-desorption. The results showed that the prepared MSN has ordered nanochannels with a pore size of 6.3, 8.1, 10.8 nm, respectively. The particle size of SRL-SMEDDS were measured by particle sizing system, which was 20.6±1.3 nm. The stirring method was developed to prepare SRL-SMEDDS-MSN. It was found that the optimal ratio of SRL-SMEDDS to MSN was 2:1, while the drug loading rate was near 0.83%, and the flow properties of SRL-SMEDDS-MSN were of good condition. The differential scanning calorimetry results proving a molecular or amorphous dispersed state of SRL in MSN while the suspension experiment has shown great reconstitution properties of SRL-SMEDDS-MSN. There is no significant influence on maximum drug release rate of different pore size of SRL-SMEDDS-MSN in 250 mL water within 2 h, while the results of the first 40 min have an obvious difference. Above all, MSN might provide a new strategy for the solidification of SMEDDS.

Flexible liposomes are an excellent drug delivery nanocarrier, however, the leakage of drugs from liposomes has become common technical obstacle in the industry and also hindered its further application seriously. It is very urgent and necessary to avoid or reduce the leakage of drugs from liposomes. In this work, five kinds of essential oils such as Folium Artemisiae Argyi oil (FA), Folium Eucalypti oil (FE), Arabian Jasmine oil (AJ), Syzygium Aromaticum oil (SA) and Fructus Forsythiae oil (FF) were encapsulated in the lipid bilayer of palmatine chloride (PC) loaded flexible nano-liposomes (PFL), then the optimal essential oil and its dosage level were determined by the external leakage curve of PC. The female Japanese white rabbits were used to evaluate the vaginal irritancy potential of liposomes samples. The pharmaceutical properties such as encapsulation efficiency, particle size, zeta potential, deformability and structure of liposomes samples were evaluated. In order to investigate the permeability of liposomes samples to deliver PC across skin and mucous membrane in vitro, the side-by-side diffusion cells were used. The results showed that the leakage of hydrosoluble PC from PFL was reduced at different degrees by the essential oils in the lipid bilayer of PFL, however, the reduction in leakage degree was obviously higher for FA than thoses of FE, AJ, SA and FF (P < 0.05), and the highest reduction in leakage degree was obtained when the FA and lipid mass ratio was 1:6. The encapsulation efficiency, particle size, zeta potential and deformability of PFL were not significantly changed after FA was encapsulated in the lipid bilayer of the PFL (P > 0.05), so did the lamellar structure of PFL. In addition, the transdermal and transmucosal permeability of PC were also enhanced obviously by encapsulating FA in the lipid bilayer of PFL, and there was no vaginal/vulvar irritation observed in the rabbits. In summary, the drug leakage was reduced by encapsulating suitable essential oil (such as FA) in the lipid bilayer of flexible liposomes, and the vaginal mucosa permeability were improved for the drug. These results provide a novel technique in the improvement of flexible nano-liposomes for drug delivery.

This study was designed to establish a multiplex allele-specific polymerase chain reaction method for simultaneous identification of Dendrobium huoshanense, D. officinale and D. devonianum, which may resolve identification problems of caulis dendrobii. Internal transcribed spacer sequences and trnL-trnF sequences of the Dendrobium species were aligned by BioEdit software, then specific SNPs of the three species were analyzed for designing allele-specific primers and the multiplex allele-specific PCR reaction system was established. The different origin of Dendrobium huoshanense, D. officinale and D. devonianum was amplified and identified by the sizes of respective band. The results showed that 584 bp, 397 bp and 211 bp bands could be amplified by D. devonianum, Dendrobium officinale and Dendrobium huoshanense respectively, when the annealing temperature was 61 ℃ and the number of cycles was 35. The limit of detection (LOD) of D. devonianum and D. huoshanense were both 1.2 ng, while D. officinale was low than 0.24 ng. The detection limit of adulterates in D. devonianum, D. devonianum and D. huoshanense mixture sample was 1%, 1% and 5% respectively. This result suggests that the method of multiplex allele-specific PCR is useful to identify D. huoshanense, D. officinale and D. devonianum is accurate and specific.

Scorpion toxin BmK AngM1 has been reported to have a strong analgesic effect. However, its anti-inflammatory activity was unknown. In this study, the recombinant BmK AngM1 (rBmK AngM1) was expressed in Escherichia coli BL21 trxB (DE3). The purified rBmK AngM1 was obtained efficiently through the IMPACT^TM-TWIN system. The anti-inflammatory activity of the recombinant protein was investigated. In order to improve the anti-inflammatory activity of rBmK AngM1, the potential active sites (Y5, Y42, R58) were substituted with different amino acids. The results showed that rBmK AngM1 and its mutants all have significant anti-inflammatory activity. The activities were significantly increased in the single mutant R58N and mutants Y5F/R58N, Y42F/R58N over the wild type protein. The data suggest that position 58 in BmK AngM1 plays a functional role in the anti-inflammatory activity. This study lays a foundation for the protein engineering design of BmK AngM1 to improve its pharmacological activity.