Structure-based drug discovery of novel fused-pyrazolone carboxamide derivatives as potent and selective AXL inhibitors

As a novel and promising antitumor target, AXL plays an important role in tumor growth, metastasis, immunosuppression and drug resistance of various malignancies, which has attracted extensive research interest in recent years. In this study, by employing the structure-based drug design and bioisosterism strategies, we designed and synthesized in total 54 novel AXL inhibitors featuring a fused-pyrazolone carboxamide scaffold, of which up to 20 compounds exhibited excellent AXL kinase and BaF3/TEL-AXL cell viability inhibitions. Notably, compound 59 showed a desirable AXL kinase inhibitory activity (IC50: 3.5 nmol/L) as well as good kinase selectivity, and it effectively blocked the cellular AXL signaling. In turn, compound 59 could potently inhibit BaF3/TEL-AXL cell viability (IC50: 1.5 nmol/L) and significantly suppress GAS6/AXL-mediated cancer cell invasion, migration and wound healing at the nanomolar level. More importantly, compound 59 oral administration showed good pharmacokinetic profile and in vivo antitumor efficiency, in which we observed significant AXL phosphorylation suppression, and its antitumor efficacy at 20 mg/kg (qd) was comparable to that of BGB324 at 50 mg/kg (bid), the most advanced AXL inhibitor. Taken together, this work provided a valuable lead compound as a potential AXL inhibitor for the further antitumor drug development.

Preclinical and early clinical studies of a novel compound SYHA1813 that efficiently crosses the blood-brain barrier and exhibits potent activity against glioblastoma

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults and is poorly controlled. Previous studies have shown that both macrophages and angiogenesis play significant roles in GBM progression, and co-targeting of CSF1R and VEGFR is likely to be an effective strategy for GBM treatment. Therefore, this study developed a novel and selective inhibitor of CSF1R and VEGFR, SYHA1813, possessing potent antitumor activity against GBM. SYHA1813 inhibited VEGFR and CSF1R kinase activities with high potency and selectivity and thus blocked the cell viability of HUVECs and macrophages and exhibited anti-angiogenetic effects both in vitro and in vivo. SYHA1813 also displayed potent in vivo antitumor activity against GBM in immune-competent and immune-deficient mouse models, including temozolomide (TMZ) insensitive tumors. Notably, SYHA1813 could penetrate the blood-brain barrier (BBB) and prolong the survival time of mice bearing intracranial GBM xenografts. Moreover, SYHA1813 treatment resulted in a synergistic antitumor efficacy in combination with the PD-1 antibody. As a clinical proof of concept, SYHA1813 achieved confirmed responses in patients with recurrent GBM in an ongoing first-in-human phase I trial. The data of this study support the rationale for an ongoing phase I clinical study (ChiCTR2100045380).

Pharmacokinetics,distribution,and excretion of sodium oligomannate,a recently approved anti-Alzheimer's disease drug in China / 药物分析学报

The National Medical Products Administration has authorized sodium oligomannate for treating mild-to-moderate Alzheimer's disease.In this study,an LC-MS/MS method was developed and validated to quantitate sodium oligomannate in different biomatrices.The plasma pharmacokinetics,tissue distri-bution,and excretion of sodium oligomannate in Sprague-Dawley rats and beagle dogs were system-atically investigated.Despite its complicated structural composition,the absorption,distribution,metabolism,and excretion profiles of the oligosaccharides in sodium oligomannate of different sizes and terminal derivatives were indiscriminate.Sodium oligomannate mainly crossed the gastrointestinal epithelium through paracellular transport following oral administration,with very low oral bioavail-ability in rats(0.6％-1.6％)and dogs(4.5％-9.3％).Absorbed sodium oligomannate mainly resided in circulating body fluids in free form with minimal distribution into erythrocytes and major tissues.So-dium oligomannate could penetrate the blood-cerebrospinal fluid(CSF)barrier of rats,showing a con-stant area under the concentration-time curve ratio(CSF/plasma)of approximately 5％.The cumulative urinary excretion of sodium oligomannate was commensurate with its oral bioavailability,supporting that excretion was predominantly renal,whereas no obvious biliary secretion was observed following a single oral dose to bile duct-cannulated rats.Moreover,only 33.7％(male)and 26.3％(female)of the oral dose were recovered in the rat excreta within 96 h following a single oral administration,suggesting that the intestinal flora may have ingested a portion of unabsorbed sodium oligomannate as a nutrient.

Design, synthesis and biological evaluation of pyrazolo3,4-

Fibroblast growth factor receptors (FGFRs) have emerged as promising targets for anticancer therapy. In this study, we synthesized and evaluated the biological activity of 66 pyrazolo[3,4-

Discovery of a series of dimethoxybenzene FGFR inhibitors with 5pyrrolo2,3-pyrazine scaffold: structure-activity relationship, crystal structural characterization and study

Genomic alterations are commonly found in the signaling pathways of fibroblast growth factor receptors (FGFRs). Although there is no selective FGFR inhibitors in market, several promising inhibitors have been investigated in clinical trials, and showed encouraging efficacies in patients. By designing a hybrid between the FGFR-selectivity-enhancing motif dimethoxybenzene group and our previously identified novel scaffold, we discovered a new series of potent FGFR inhibitors, with the best one showing sub-nanomolar enzymatic activity. After several round of optimization and with the solved crystal structure, detailed structure-activity relationship was elaborated. Together with metabolic stability tests and pharmacokinetic profiling, a representative compound () was selected and tested in xenograft mouse model, and the result demonstrated that inhibitor was effective against tumors with FGFR genetic alterations, exhibiting potential for further development.

Targeting ERK, an Achilles' Heel of the MAPK pathway, in cancer therapy

The mitogen-activated protein kinases (MAPK) pathway, often known as the RAS-RAF-MEK-ERK signal cascade, functions to transmit upstream signals to its downstream effectors to regulate physiological process such as cell proliferation, differentiation, survival and death. As the most frequently mutated signaling pathway in human cancer, targeting the MAPK pathway has long been considered a promising strategy for cancer therapy. Substantial efforts in the past decades have led to the clinical success of BRAF and MEK inhibitors. However, the clinical benefits of these inhibitors are compromised by the frequently occurring acquired resistance due to cancer heterogeneity and genomic instability. This review briefly introduces the key protein kinases involved in this pathway as well as their activation mechanisms. We also generalize the correlations between mutations of MAPK members and human cancers, followed by a summarization of progress made on the development of small molecule MAPK kinases inhibitors. In particular, this review highlights the potential advantages of ERK inhibitors in overcoming resistance to upstream targets and proposes that targeting ERK kinase may hold a promising prospect for cancer therapy.

Tetrahydroisoquinolines as novel histone deacetylase inhibitors for treatment of cancer

Histone acetylation is a critical process in the regulation of chromatin structure and gene expression. Histone deacetylases (HDACs) remove the acetyl group, leading to chromatin condensation and transcriptional repression. HDAC inhibitors are considered a new class of anticancer agents and have been shown to alter gene transcription and exert antitumor effects. This paper describes our work on the structural determination and structure-activity relationship (SAR) optimization of tetrahydroisoquinoline compounds as HDAC inhibitors. These compounds were tested for their ability to inhibit HDAC 1, 3, 6 and for their ability to inhibit the proliferation of a panel of cancer cell lines. Among these, compound 82 showed the greatest inhibitory activity toward HDAC 1, 3, 6 and strongly inhibited growth of the cancer cell lines, with results clearly superior to those of the reference compound, vorinostat (SAHA). Compound 82 increased the acetylation of histones H3, H4 and tubulin in a concentration-dependent manner, suggesting that it is a broad inhibitor of HDACs.

Chromopeptide A, a highly cytotoxic depsipeptide from the marine sediment-derived bacterium Chromobacterium sp. HS-13-94

A bicyclic depsipeptide, chromopeptide A (1), was isolated from a deep-sea-derived bacterium Chromobacterium sp. HS-13-94. Its structure was determined by extensive spectroscopic analysis and by comparison with a related known compound. The absolute configuration of chromopeptide A was established by X-ray diffraction analysis employing graphite monochromated Mo K α radiation (λ=0.71073 Å) with small Flack parameter 0.03. Chromopeptide A suppressed the proliferation of HL-60, K-562, and Ramos cells with average IC50 values of 7.7, 7.0, and 16.5 nmol/L, respectively.

The potential molecular targets of marine sulfated polymannuroguluronate interfering with HIV-1 entry. Interaction between SPMG and HIV-1 rgp120 and CD4 molecule.

The potential targets of marine sulfated polymannuroguluronate (SPMG) involved in inhibition of HIV-1 entry were investigated by surface plasmon resonance and flow cytometry. Results indicated that binding of SPMG either to soluble oligomeric rgp120 or to complexed rgp120-sCD4 mainly resided in V3 loop region. In addition, SPMG was shown to be less accessible for sCD4 when sCD4 had pre-interacted with rgp120, though SPMG per se multivalently bound to sCD4 with relatively low affinity. While the pre-incubation of SPMG with rgp120 caused a partial blockade of rgp120 binding to sCD4, suggesting that SPMG either shared common binding sites on gp120 with sCD4 or masked the docking sites of gp120 for sCD4. Taken together, V3 domain was demonstrated to be the major site mediating interaction of SPMG with complexed rgp120-sCD4. It seems likely that SPMG binds to both rgp120 and sCD4, but has less accessibility for sCD4 when sCD4 has already bound to rgp120. Nevertheless, addition of SPMG either prior to or after the interaction of rgp120 with sCD4 may suppress rgp120 binding to sCD4. The exact pattern of this trimolecular complex formation at the cell membrane-anchored virus level requires further clarification.

Progress in soluble A? oligomers in Alzheimer's disease and drugs targeting A? oligomers / 中国药理学通报

A? plays a crucial role in Alzheimer's Disease and the soluble A? oligomers have been recognized as the emerging neurotoxins,which ultimately cause memory impairment and neuronal loss through different mechanisms.The development of novel drugs targeting A? oligomers indicates new and promising therapy approaches for AD.The pathological roles as the proximal toxins in AD and the compounds,targeting soluble A? oligomers,which are currently in preclinical and early clinical development,are reviewed.

The signal transduction and drug development of epithelial-mesenchymal transition / 中国药理学通报

Epithelial-mesenchymal-transition(EMT) plays a key role in the formation of embryo.At present,it is believed that EMT also contribute to the metastasis of primary tumors.A lot of signaling pathways participate in EMT,such as Wnt/?-catenin,Notch,Hedgehog,TGF? and growth factor receptors.The drugs that affect these pathways may play an important part in oncotherapy.

Glycosylations and cancer cell cycle / 中国药理学通报

It is well known that oligosaccharides in glycoproteins and glycolipids play crucial roles in a variety of cellular functions, such as proliferation, differentiation, and intercellular communications. The oligosaccharides of cell are increasingly being recognized as one of the most prominent biochemical alterations associated with malignant transformation and tumorigenesis. Glycosylations in different cell cycle and cell growth periods are significantly distinct, and these differences affect the cell cycle progression. In the past decades, along with the advances in genomics and proteomics, the functional significance of cancer-associated changes in glycosylation has been revealed. Eukaryotic organisms depend on an intricate and evolutionary conserved cell cycle to control cell devision. Mistakes in cell cycle process lead to cancer. This review highlights the relations between cell cycle and glycosylation changes in cancer cells.

Integrin and tumor metastasis / 中国药理学通报

Integrin is an important cellular adhesion molecule,which mediates many biologic actions such as cell-cell adhesion or cell-extracellular matrix adhesion.The relationship between integrin and tumor metastasis,as well as the important role of integrin during tumor progress,is reviewed in this article,which gives us the theoretical base for new pathways of cancer treatment.

Investigation progress on receptors of saccharides / 中国药理学通报

More and more interest has been put on the investigations of saccharides recent years. Saccharides have a vast number of bioactivities and the immunomodulating effect of saccharides is one of the important mechanisms via which saccharides exert their bioactivities. Saccharides exert their bioactivities mainly by interacting with all kinds of saccharide receptors on cells, followed by a series of signal transduction process. In the present paper, numerous saccharide receptors and the characters of the mutual interaction between saccharides and their receptors are summarized.

Construction of PSA and NCAM differently expressing COS-7 cell line and the effects of PSA on migration and invasion of the diverse COS-7 cell / 中国药理学通报

Aim To construct a polysialic acid(PSA)and neural cell adhesion molecule(NCAM)differently expressing COS-7 cell line,and investigate the effects of PSA on the cell adhesion,migration and invasion,aiming to establish the base for further investigation of the signaling passway of the diverse celluar migration and invasion,and elucidate the molecular mechanism of PSA promoting cancer cell metastasis.Methods A polysialic acid and neural cell adhesion molecule differently expressing COS-7 cell line was constructed by transient cotransfection,and the cotransfection efficiency was determined by Western blot and flow cytometry.Adhesion assay was used to investigate the effect of PSA on cell adhesion ability;transwell assay was used to measure migration and invasion ability.Results The PSA and NCAM differently expressing COS-7 cell line was successfully constructed,which demonstrated PSA inhibited the cell adhesion to basement membrane,and promoted the migration and invasion ability.Conclusions The constructed polysialic acid and neural cell adhesion molecule differently expressing COS-7 cell line can be used to investigate molecular mechanism of promoting cancer cell metastasis induced by PSA in the future.

Polysaccharide sulfate 916 inhibits neutrophil-endothelial adhesion / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion.</p><p><b>METHODS</b>Cell adhesion was evaluated by testing neutrophil myeloperoxidase activity. Expression of adhesion molecule in human umbilical vein endothelial cell (HUVEC) was measured by ELISA. The neutrophil activation rate induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was tested by nitroblue tetrazolium (NBT) reduction.</p><p><b>RESULTS</b>Tumor necrosis factor alpha (TNFalpha, 50 - 800 U/ml) increased the adherence of neutrophil to TNFalpha-stimulated HUVEC in a concentration and time dependent manner. PS916 (0.01 - 1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFalpha-stimulated HUVEC. fMLP increased the activation rate of neutrophils independent of concentration. PS916 also inhibited the adherence of fMLP-activated neutrophils to HUVEC. Moreover, PS916 inhibited adhesion molecule expression in TNFalpha-stimulated HUVEC.</p><p><b>CONCLUSIONS</b>PS916 inhibited neutrophil-endothelial adhesion. The mechanism of its action was partially related to suppressing the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1).</p>

Effect of polysaccharide sulfate 916 on the production of nitric oxide in ECV304 cells induced by cytokines and H(2)O(2) / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of polysaccharide sulfate 916 (PS916) on the production of nitric oxide (NO) in ECV304 cells induced by tumor necrosis factor-alpha (TNF alpha), interleukin-1 beta (IL-1 beta) and H(2)O(2) in vitro.</p><p><b>METHODS</b>Production of NO in ECV304 cells was measured by the Griess method and the proliferation of cells was tested by the MTT method. The activity of NO synthase was detected spectrophotometrically.</p><p><b>RESULTS</b>Production of NO in ECV304 cells decreased after treatment with 40 ng/ml IL-1 beta and 40 ng/ml TNF alpha, but increased in the presence of H(2)O(2) 0.1 mmol/L. PS916 significantly enhanced NO production in ECV304 cells in a dose-dependent manner in the TNF alpha and IL-1 beta treated groups and decreased it in the H(2)O(2) treated group. Proliferation of ECV304 cells was inhibited by TNFalpha and H(2)O(2) and no effect was found in the IL-1 beta treated group. PS916 increased the proliferation of cells treated with TNFalpha and H(2)O(2) dose-dependently. In vitro, PS916 has no effect on the activity of NO synthase.</p><p><b>CONCLUSION</b>PS916 has a protective effect on ECV304 cells exposed to IL-1 beta, TNF alpha and H(2)O(2).</p>

INHIBITORY EFFECTS OF D-POLYMANNURONIC SULFATE ON PROLIFERATION OF RAT VASCULAR SMOOTH MUSCLE CELLS AND ITS RELATED MECHANISMS OF ACTION IN VITRO / 药学学报

AIM　To investigate the inhibitory effects of D-polymannuronic sulfate (DPS) on the proliferation of rat vascular smooth muscle cells (VSMC) induced by basic fibroblast growth factor (bFGF) or interleukin-1 (IL-1) and its related mechanisms. METHODS　Rat aortic smooth muscle cells pretreated with DPS in concentrations ranging from 0.001 μg*mL-1 up to 100 μg*mL-1 were incubated at 37℃ for 24 h, followed by addition of bFGF (50 ng*mL-1) or IL-1 (50 U*mL-1) for another 24 h. The effects of DPS on the proliferation of VSMC were evaluated by MTT assays. VSMC were pretreated with DPS in concentrations ranging from 0.001 μg*mL-1 up to 1 μg*mL-1, followed by addition of L-NAME (0.1 μg*mL-1) or bFGF (50 ng*mL-1) for 24 h. Supernatant nitric oxide (NO) was determined with NO assay kit, while supernatant angiotensin II (Ang II) and endothelin-1 (ET-1) were measured by radioimmunoassay. RESULTS　DPS exerted antiproliferative effects at concentrations ranging from 0.01 μg*mL-1 to 10 μg*mL-1, and its maximal effect was observed at the concentration of 1 μg*mL-1. Also, the suppressing actions of DPS on the proliferation of VSMC were diminished by increasing the concentrations of bFGF or IL-1. Furthermore, DPS increased NO synthesis and decreased Ang II and ET-1 contents released from VSMC in a concentration-dependent manner. CONCLUSION　DPS afforded the antiproliferative effects on bFGF- or IL-1-treated VSMC and its underlying mechanisms were associated with enhancement of NO synthesis and decrement of Ang II and ET-1 production/release in vitro.

The relationship between sulfated polysaccharides and atheroscle rosis / 中国药理学通报

The atherogensis was involved in a complex pathological process. Injury to endothelial cells of blood vessels was confirmed to be the in itial stage of this process. Migration to subendothelial layer and accumulation and proliferation of smooth muscle cells were attributed to various cytokines an d adhesive molecules secreted by activated endothelial cells, subsequently resul ting in aggregation of lymphocytes,platelets, monocytes and macrophages in the i ntima of artery. These cellular components ultimatedly led to the formation of a mature atherosclerotic plaque. Its quite acknowledged that a better understan ding of the atherogenic events might promise us the development of new chemical entities of anti-atherosclerotic therapies. A large body of evidence has demons trated that sulfated polysaccharides played a critical role in the development o f atherosclerosis. The underlying mechanisms of the anti-atherosclerotic activi ty of sulfated polysaccharides were reported to contribute to protecting against endothelial cells injury,inhibiting migration and proliferation of vascular smo oth muscle cells, and reducing the adhesion of inflammatory cells, platelets and lymphocytes. And also, the prevention of complement activation by sulfated poly saccharides could not be excluded. On the other hand,the promoting effects of su lfated polysaccharides atherosclerosis was also reported. Its therefore conclu ded that the relationships between atheriosclerosis and sulfated polysaccharides remained to be further elucidated.

Binding experiments of 911-FITC with Immunocytes / 中国海洋药物

Lymphocytes from thymus, spleen, macrophages and endothelial cells w.ere incubated with the fluorescent probe of 911-FITC and the colouring method with flow cytometry and fluorescent microscope was reviewed in this paper. The results indicated that 911-FITC could specifically bind to lymphocytes, macrophages and endothelial cells. These findings suggested that 911 could play a immunomodulatory effect by direct combination with immu-nocytes.