ABSTRACT
We designed and synthesized eighteen lycorine derivatives with five different structural types, and evaluated their antiviral activities on a HCoV-OC43-infected H460 cell model. Structure-activity relationships suggested that the introduction of appropriate substituents on the 6N atom of lycorine was beneficial to activity. Compound 6a gave a good activity with the half effective concentration (EC50) and selectivity index (SI) values of 2.36 μmol·L-1 and 16.52, respectively. Surface plasmon resonance (SPR) result indicated that 6a might target the non-structural protein 12 (NSP12) subunit in RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with the dissociation constant (KD) value of 1.36 μmol·L-1. Molecular docking indicated that 6a might act on nidovirus RdRp-associated nucleotidyltransferase (NiRAN) catalytic center of NSP12, distinct from the mechanism of nucleoside-like drugs such as remdesivir. This study provides scientific data for the development of lycorine derivatives into a new class of anti-SARS-CoV-2 small molecule inhibitors.
ABSTRACT
Chimeric antigen receptor T-cell (CAR-T) immunotherapy has achieved good efficacy in treatment of hematological malignancies. As a precise and individualized treatment method, CAR-T is gradually moving towards commercialization. In addition to the introduction of corresponding policies and guiding principles, the related detection protocols should also be updated and improved to maximize its effect and achieve precise individualization. This article introduces and expands the concept of "companion diagnostics" that first appeared in targeted drugs, and introduces the significances of various detection technologies and biomarkers for patient screening, safety monitoring and evaluation of efficacy and CAR-T function in the whole process of CAR-T treatment.
ABSTRACT
Objective: Murine CD19 chimeric antigen receptor T-cell (CAR-T) products have been approved for the treatment of refractory/relapsed (R/R) B-cell acute lymphocytic leukemia (B-ALL) ; moreover, humanized products are also undergoing clinical trials. This study aimed to explore the differences in safety and short- and long-term follow-up efficacy between humanized and murine CD19 CAR-T-cells for treating relapsed and refractory B-ALL. Methods: Clinical data of 80 patients with R/R B-ALL treated with CD19-targeted CAR-T-cells at the Union Hospital of Tongji Medical College of Huazhong University of Science and Technology between May 2016 and March 2023 were analyzed, which included 31 patients with murine CAR-T and 49 with humanized products. Results: The proportion of patients with cytokine-release syndrome (CRS) in the murine and humanized groups was 63.1% and 65.3%, respectively. Moreover, a higher proportion of patients suffered from severe CRS in the murine group than in the humanized CAR-T group (19.4% vs 8.2%, P=0.174). Furthermore, one patient per group died of grade 5 CRS. The incidence of grade 1-2 immune effector cell-associated neurotoxicity syndrome (ICANS) was 12.9% and 6.1%, respectively; severe ICANS were not observed. Among patients receiving murine CAR-T-cells, an overall response (OR) was observed in 74.2%. Conversely, the OR rate of patients receiving humanized CAR-T-cells was 87.8%. During the median follow-up time of 10.5 months, the median recurrence-free survival (RFS) of patients with murine CAR-T-cells was 12 months, which was as long as that of patients with humanized CAR-T-cells. The median overall survival (OS) were not reached in both groups. Of the 45 patients with a bone marrow burden over 20% at baseline, humanized CAR-T therapy was associated with a significantly improved RFS (43.25% vs 33.33%, P=0.027). Bridging transplantation was an independent factor in prolonging OS (χ(2)=8.017, P=0.005) and PFS (χ(2)=6.584, P=0.010). Common risk factors, such as age, high proportion of bone marrow blasts, and BCR-ABL fusion gene expression, had no significant effect on patients' long-term follow-up outcomes. Three patients reached complete remission after reinfusion of humanized CAR-T-cells. However, one patient relapsed one month after his second infusion of murine CAR-T-cells. Conclusions: The results indicate that humanized CAR-T therapy showed durable efficacy in patients with a higher tumor burden in the bone marrow without any influence on safety. Moreover, it could overcome immunogenicity-induced CAR-T resistance, providing treatment options for patients who were not treated successfully with CAR-T therapies.
Subject(s)
Animals , Humans , Mice , Antigens, CD19 , Burkitt Lymphoma/drug therapy , Cell- and Tissue-Based Therapy , Follow-Up Studies , Immunotherapy, Adoptive , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Receptors, Chimeric AntigenABSTRACT
Immunotherapy has been one of the most promising approaches in tumor-treating fields, including immune checkpoint inhibitors, bispecific antibodies, and CAR-T therapy, etc. It has achieved major breakthroughs in the treatment of hematological and other malignancies. However, related safety management issues are becoming increasingly prominent, especially the diagnosis and treatment of coagulopathy deserves the attention of clinical and laboratory physicians. Therefore, this review summarizes immunotherapy-related coagulopathy from the perspectives of epidemiology, pathogenesis and laboratory indicators and provides guidance for early clinical identification, diagnosis and intervention.
ABSTRACT
Chimeric antigen receptor T-cell (CAR-T) has made a breakthrough in the treatment of hematologic malignancies, but drug resistance and recurrence have limited its wide application. And at the 63rd annual meeting of the American Society of Hematology, a series of related reports on the mechanism and prevention strategies of drug resistance and recurrence after CAR-T therapy were carried out. These reports provide important indications for improving the clinical efficacy of CAR-T therapy and reducing relapse.
ABSTRACT
Chimeric antigen receptor T cells (CAR-T) therapy is progressing rapidly, and its safety has been widely concerned. At the 62nd American Society of Hematology Annual Meeting, a series of reports on the mechanism, predictive indicators and treatment strategies of major adverse reactions were carried out. These reports have certain guiding significance for comprehensively improving the safety of CAR-T therapy.
ABSTRACT
Abstract Chimetic antigen receptor T cells (CAR-T) are a kind of novel killer cells derived from autogenous or allogeneic T cells targeting the tumor specific antigens by gene engineering transpormation. The CAR-T cells possess the advantages of high specificity, high efficiency and non-MHC restriction, thus achieving good therapeutic effects in relapsed/refractory hematological tumors and some solid tumors. The process of CAR-T cell preparation includes cell isolation and purification, activation and differentiation, gene modification, in vitro amplification, phenotypic quality control, preservation and reinfusion. In recent years, with the rapid development of flow cytometry, this technology has been widely used in all aspects of CAR-T cell therapy, including pre-treatment screening, in vitro preparation and post-transfusion monitoring, which plays an important role in its innovative optimization.
ABSTRACT
Thrombosis and hemostasis are clinical interdisciplinary subjects involving multiple specialties. Related thrombus and hemorrhagic diseases seriously endanger health. Since the founding of the People's Republic of China, the diagnosis and treatment level of thrombus and hemorrhagic diseases in China have been continuously improved, the theoretical research has been continuously deepened, and a series of fruitful results have been achieved in the basic and clinical research on platelets, coagulation factors, anticoagulant and fibrinolytic systems. This paper summarizes the current situation and future development direction of related representative work.
ABSTRACT
The efficacy and safety of recombinant tissue plasminogen activator (rtPA) need to be improved due to its low bioavailability and requirement of large dose administration.The purpose of this study was to develop a fibrin-targeted nanoparticle (NP) drug delivery system for thrombosis combination therapy.We conjugated rtPA to poly(ethylene glycol)-poly(ε-caprolactone) (PEG-PCL) nanoparticles (rtPA-NP) and investigated its physicochemical characteristics such as particle size,zeta potential,enzyme activity of conjugated rtPA and its storage stability at 4℃.The thrombolytic activity of rtPA-NP was evaluated in vitro and in vivo as well as the half-life of rtPA-NP,the properties to fibrin targeting and its influences on systemic hemostasis in vivo.The results showed that rtPA-NP equivalent to 10％ of a typical dose of rtPA could dissolve fibrin clots and were demonstrated to have a neuroprotective effect after focal cerebral ischemia as evidenced by decreased infarct volume and improved neurological deficit (P＜0.001).RtPA-NP did not influence the in vivo hemostasis or coagulation system.The half-life of conjugated rtPA was shown to be approximately 18 times longer than that of free rtPA.These experiments suggested that rtPA-conjugated PEG-PCL nanoparticles might be a promising fibrin-targeted delivery system for a combination treatment of thrombosis.
ABSTRACT
The efficacy and safety of recombinant tissue plasminogen activator (rtPA) need to be improved due to its low bioavailability and requirement of large dose administration.The purpose of this study was to develop a fibrin-targeted nanoparticle (NP) drug delivery system for thrombosis combination therapy.We conjugated rtPA to poly(ethylene glycol)-poly(ε-caprolactone) (PEG-PCL) nanoparticles (rtPA-NP) and investigated its physicochemical characteristics such as particle size,zeta potential,enzyme activity of conjugated rtPA and its storage stability at 4℃.The thrombolytic activity of rtPA-NP was evaluated in vitro and in vivo as well as the half-life of rtPA-NP,the properties to fibrin targeting and its influences on systemic hemostasis in vivo.The results showed that rtPA-NP equivalent to 10％ of a typical dose of rtPA could dissolve fibrin clots and were demonstrated to have a neuroprotective effect after focal cerebral ischemia as evidenced by decreased infarct volume and improved neurological deficit (P＜0.001).RtPA-NP did not influence the in vivo hemostasis or coagulation system.The half-life of conjugated rtPA was shown to be approximately 18 times longer than that of free rtPA.These experiments suggested that rtPA-conjugated PEG-PCL nanoparticles might be a promising fibrin-targeted delivery system for a combination treatment of thrombosis.
ABSTRACT
This study examined the association of a common polymorphic allele (25G) of the low-density lipoprotein receptor-related protein1 (LRP1) gene with myocardial infarction (MI). The genotypes of LRP1 25CG (rs35282763) were determined in 347 MI patients and 347 age- and sex-frequency-matched controls from an unrelated Chinese Han population. Factor VIII (FVIII) levels were measured in the MI patients and controls by chromogenic assay and enzyme-linked immunosorbent assay (ELISA). The results showed that LRP1 25CG (rs35282763) genotype distribution did not differ significantly between patients (n=206 for 25CC, n=122 for 25CG) and controls (n=191 for 25CC, n=126 for 25CG; P>0.05). The 25G allele was not associated with a reduced risk of MI (P>0.05). Further stratifications for age, sex, and other cardiovascular risk factors did not affect the negative findings. It was concluded that the presence of the G allele at the 25CG (rs35282763) polymorphism of the LRP1 is not associated with a reduced risk of MI, and genotyping for LRP1 25CG (rs35282763) polymorphism is not useful in assessing the individual risk of MI.
ABSTRACT
DNA repair processes play a role in the development of drug resistance which represents a huge obstacle to leukemia chemotherapy. Histone H2AX phosphorylation (ser139) (γH2AX) occurs rapidly at the onset of DNA double strand break (DSB) and is critical to the regulation of DSB repair. If DNA repair is successful, cells exposed to anti-neoplastic drugs will keep entering the cycle and develop resistance to the drugs. In this study, we investigated whether γH2AX can be used as an indicator of tumor chemosensitivity and a potential target for enhancing chemotherapy. K562 and multi-drug resistant cell line K562/A02 were exposed to adriamycin (ADR) and γH2AX formed. Flow cytometry revealed that percentage of cells expressing γH2AX was increased in a dose-dependent manner and the percentage of K562/A02 cells was lower than that of K562 cells when treated with the same concentration of ADR. In order to test the potential of γH2AX to reverse drug resistance, K562/A02 cells were treated with PI3K inhibitor LY294002. It was found that LY249002 decreased ADR-induced γH2AX expression and increased the sensitivity of K562/A02 cells to ADR. Additionally, the single-cell gel electrophoresis assay and the Western blotting showed that LY249002 enhanced DSBs and decreased the expression of repair factor BRCA1. These results illustrate chemosensitivity can partly be measured by detecting γH2AX and drug resistance can be reversed by inhibiting γH2AX.
ABSTRACT
The binding function of EGF1 domain peptide with tissue factor (TF) and its ability of triggering coagulation were explored. The TF expression model in vitro was established by lipopolysaccharide induction. The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry (FCM). The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance (SPR). The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay. The FCM results showed recombinant factor VII (rFVII) could definitely depress the integration of EGFP-EGF1 with recombinant TF (rTF) (68.65%+/-3.86% vs 57.98%+/-4.71%, P<0.01). The SPR results indicated the association constant ka of EGFP-EGF1 proteins was higher than rFVII (8.29+/-1.39 vs 3.75+/-0.32, P<0.01). However, the EGFP-EGF1 protein lost the activity of triggering coagulation as compared with blood plasma of normal SD rats (56.8+/-3.2 s vs 17.8+/-3.4 s, P<0.01). It was concluded that the rat EGF1 peptide could specifically bind to TF without the ability of triggering coagulation. EGF1 peptide may be a good target head for delivering drugs to TF in anticoagulation therapy.