ABSTRACT
Glycosite-specific antibody‒drug conjugatess (gsADCs), harnessing Asn297 N-glycan of IgG Fc as the conjugation site for drug payloads, usually require multi-step glycoengineering with two or more enzymes, which limits the substrate diversification and complicates the preparation process. Herein, we report a series of novel disaccharide-based substrates, which reprogram the IgG glycoengineering to one-step synthesis of gsADCs, catalyzed by an endo-N-acetylglucosaminidase (ENGase) of Endo-S2. IgG glycoengineering via ENGases usually has two steps: deglycosylation by wild-type (WT) ENGases and transglycosylation by mutated ENGases. But in the current method, we have found that disaccharide LacNAc oxazoline can be efficiently assembled onto IgG by WT Endo-S2 without hydrolysis of the product, which enables the one-step glycoengineering directly from native antibodies. Further studies on substrate specificity revealed that this approach has excellent tolerance on various modification of 6-Gal motif of LacNAc. Within 1 h, one-step synthesis of gsADC was achieved using the LacNAc-toxin substrates including structures free of bioorthogonal groups. These gsADCs demonstrated good homogeneity, buffer stability, in vitro and in vivo anti-tumor activity. This work presents a novel strategy using LacNAc-based substrates to reprogram the multi-step IgG glycoengineering to a one-step manner for highly efficient synthesis of gsADCs.
ABSTRACT
Aim To discuss the influence of endosome/ lysosome transport proteins SNX10 on macrophage, providing new potential targets for the treatment of variety of related immune diseases . Methods The genotype of mice was identified by PCR. The role of SNX10 in phagocytosis of bacterial components and sterilization by macrophages were assessed. The levels of TNF-α、IL-12 / 23 p40 and IL-6 were measured by q-PCR and ELISA assay. Finally, the NF-κB signaling pathway was evaluated by Western blot and immuno-fluorescence staining. Results Ex vivo experiments showed that SNX10 knockout could enhance bactericid-al ability and inhibit the expression and production of TNF-α, IL-12 / 23 p40 and IL-6 of macrophages.These effects might attribute to the inhibition of NF-κB signaling pathway activation. Conclusion SNX10 knockout could enhance bactericidal ability and inhibit the inflammatory response of macrophages, and its mechanism may be achieved through the NF-κB signa-ling pathway.
ABSTRACT
Objective To investigate the clinical effect and the prospect of Infliximab in treatment of intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients.Methods Clinical features,inflammatory markers and coronary changes were observed in 2 cases of IVIG-resistant KD patients hospitalized in Peking University First Hospital,who were treated effectively by Infliximab.Relevant researches on the mechanism and progress of the Infliximab treatment for IVIG-resistant KD in the last 10 years were reviewed at the same time.Results Two KD patients hospitalized in Peking University First Hospital had been treated with 2 g/kg IVIG for 2 times and followed by methylprednisolone treatment.However,fever and other clinical manifestations occurred again after 2 days and 6 days when temperature returned normal.They both defervesced and all the symptoms were improved after 1 dose of Infliximab (5 mg/kg) by laboratory examinations.Four published literatures of the basic research and 9 retrospective or prospective clinical researches of Infliximab treatment of KD showed that Infliximab alleviated the inflammatory level in the KD patients significantly.Complete remission was up to 72.73％-92.11％.Those KD patients defervesced within 12 h,with dramatic improvement of symptoms and signs.Arthralgia also disappeared in the patients with arthritis.Only 1 case was complicated with hepatitis in the acute phase and cholecystitis in recovery time.A phase 3 randomised,double-blinded,placebo-controlled trial had been done to assess the addition of Infliximab to the standard therapy.Conclusions Infliximab is a feasible choice for IVIG-resistant KD patients.Efficacy and safety of Infliximab for KD treatment have been proved in the literature.However,Infliximab for KD treatment has not been indicated in the drug instruction,so the informed consent from the guardians and Ethics Committee is needed.
ABSTRACT
Objective To assess the wall elasticity of the common carotid artery using echo-tracking in healthy children and children with hyperlipidemia.Methods The study included 68 normal healthy children and 25 children with hyperlipidemia.All the subjects were 3-18 years old.They were divided into three groups according to ages 3-6 years old group,7-13 years old group and 14-18 years old group.Echo-tracking was used to measure the pressure-strain elastic modulus (E?),stiffness parameter(?)and arterial compliance (AC) of the common carotid artery.Results The average values of E? were different among the three age groups in normal healthy children (all P