Design, Synthesis, and Structure-Activity Relationship of Novel Pyridazinone-Based PARP7/HDACs Dual Inhibitors for Elucidating the Relationship between Antitumor Immunity and HDACs Inhibition.

Histone deacetylases (HDACs) inhibitors such as vorinostat (SAHA) has been used to treat hematologic malignancies (rather than solid tumors) and have been found to suppress the JAK/STAT, a critical signal pathway for antitumor immunity, while PARP7 inhibitor RBN-2397 could activate the type I interferons (IFN-I) pathway, facilitating downstream effects such as STAT1 phosphorylation and immune activation. To elucidate whether simultaneous inhibition of these two targets could interfere with these two signal pathways, a series of pyridazinone-based PARP7/HDACs dual inhibitors have been designed, synthesized, and evaluated in vitro and in vivo experiments. Compound 9l was identified as a potent and balanced dual inhibitor for the first time, exhibiting excellent antitumor capabilities both in vitro and in vivo. This suggests that 9l can be used as a valuable tool molecule for investigating the relationship between anticancer immunity and HDAC inhibition.

Design, synthesis and biological evaluation of novel 1,2,4a,5-tetrahydro-4H-benzo[b][1,4]oxazino[4,3-d][1,4]oxazine-based AAK1 inhibitors with anti-viral property against SARS-CoV-2.

Coronavirus entry into host cells hinges on the interaction between the spike glycoprotein of the virus and the cell-surface receptor angiotensin-converting enzyme 2 (ACE2), initiating the subsequent clathrin-mediated endocytosis (CME) pathway. AP-2-associated protein kinase 1 (AAK1) holds a pivotal role in this pathway, regulating CME by modulating the phosphorylation of the µ subunit of adaptor protein 2 (AP2M1). Herein, we report a series of novel AAK1 inhibitors based on previously reported 1,2,4a,5-tetrahydro-4H-benzo[b] [1,4]oxazino[4,3-d] [1,4]oxazine scaffold. Among 23 synthesized compounds, compound 12e is the most potent one with an IC50 value of 9.38 ± 0.34 nM against AAK1. The in vitro antiviral activity of 12e against SARS-CoV-2 was evaluated using a model involving SARS-CoV-2 pseudovirus infecting hACE2-HEK293 host cells. The results revealed that 12e was superior in vitro antiviral activity against SARS-CoV-2 entry into host cells when compared to SGC-AAK1-1 and LX9211, and its activity was comparable to that of a related and reference compound 8. Mechanistically, all AAK1 inhibitors attenuated AAK1-induced phosphorylation of AP2M1 threonine 156 and disrupted the direct interaction between AP2M1 and ACE2, ultimately inhibiting SARS-CoV-2 infection. Notably, compounds 8 and 12e exhibited a more potent effect in suppressing the phosphorylation of AP2M1 T156 and the interaction between AP2M1 and ACE2. In conclusion, novel AAK1 inhibitor 12e demonstrates significant efficacy in suppressing SARS-CoV-2 infection, and holds promise as a potential candidate for developing novel antiviral drugs against SARS-CoV-2 and other coronavirus infections.

Recent progress in discovery of novel AAK1 inhibitors: from pain therapy to potential anti-viral agents.

Adaptor associated kinase 1 (AAK1), a member of the Ark1/Prk1 family of Ser/Thr kinases, is a specific key kinase regulating Thr156 phosphorylation at the µ2 subunit of the adapter complex-2 (AP-2) protein. Due to their important biological functions, AAK1 systems have been validated in clinics for neuropathic pain therapy, and are being explored as potential therapeutic targets for diseases caused by various viruses such as Hepatitis C (HCV), Dengue, Ebola, and COVID-19 viruses and for amyotrophic lateral sclerosis (ALS). Centreing on the advances of drug discovery programs in this field up to 2023, AAK1 inhibitors are discussed from the aspects of the structure-based rational molecular design, pharmacology, toxicology and synthetic routes for the compounds of interest in this review. The aim is to provide the medicinal chemistry community with up-to-date information and to accelerate the drug discovery programs in the field of AAK1 small molecule inhibitors.

Design, Synthesis and Biological Evaluation of Histone Deacetylase Inhibitors Based on Pyrrolo[2,3-d]pyrimidine and Pyrrolo[2,3-b]pyridine Scaffolds.

Histone deacetylases (HDACs) are validated targets for the development of anticancer drugs in epigenetics. We have designed and synthesized a series of novel HDAC inhibitors based on pyrrolo[2,3-d]pyrimidine and pyrrolo[2,3-b]pyridine scaffolds. Compound B3 {(E)-3-(4-(((1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-yl)amino)methyl)phenyl)-N-hydroxyacrylamide} exhibits potent inhibitory activity against HDACs 1, 2, 3, 6, and 8 with IC50 values of 5.2, 6.0, 8.8, 4.4, and 173.0 nM, respectively. It exhibited potent antiproliferative effects against three tumour cell lines (IC50 values of 0.13, 0.37, and 1.11 µM, against MV-4-11, K562, and WSU-DLCL-2 cells, respectively) with two- to sixfold improvement relative to suberoylanilide hydroxamic acid (SAHA). Mechanistic studies on WSU-DLCL-2 cells revealed that B3 exhibits anticancer effects through the induction of G0 /G1 -phase arrest and promotion of apoptosis. The results of this study warrant further investigation of this compound series for the treatment of hematological malignancy.

Recent progress in nickel-catalyzed carboboration of alkenes.

Alkenes represent one of the most useful building blocks for organic synthesis, owing to their abundance and versatile reactivity. Transition metal (Pd, Cu, Co, Ni, Fe, etc.) catalyzed difunctionalization of alkenes provides efficient access to substituted molecules from readily available alkenes by installing functional groups across their carbon-carbon double bonds. Particularly, Nickel-based catalytic complexes have attracted a great deal of attention. This is because they are prone to undergoing oxidative addition and slow ß-hydride elimination, and can access both two-electron and radical pathways. Numerous elegant Ni-catalyzed cross-coupling methods, e.g., (hetero)arylboration, alkenylboration, alkylboration and alkynylboration of alkenes, have been developed with broad scopes and a high tolerance to a variety of functional groups. Therefore, the Ni-catalyzed carboboration of alkenes has become an efficient synthetic protocol to deliver substituted compounds by the cross-coupling of alkenes, electrophiles, and B2Pin2. Despite this progress, a number of challenging issues remaining in the field include broadening the types of carboboration reactions, especially the asymmetric ones, diversifying electrophile types (which is limited to halogens for now) and gaining profound insight into the reaction mechanisms. This review summarizes the recent progress in this emerging field from the literature published since 2018. It will provide the scientific community with convenience to access collective information and to accelerate their further research in order to broaden the scope of methodology and application in drug discovery programs.

Novel hydroxyl carboximates derived from ß-elemene: design, synthesis and anti-tumour activities evaluation.

A series of novel N-alkyl-N-hydroxyl carboximates derived from ß-elemene were fortuitously discovered. Most of them showed more potent anti-proliferative activities than their lead compound ß-elemene (1). Notably, compound 11i exhibited significant inhibitory effects on the proliferation of three lung cell lines (H1975, A549 and H460) and several other tumour cell lines (H1299, U87MG, MV4-11, and SU-DHL-2). Preliminary mechanistic studies revealed that compound 11i could significantly induce cell apoptosis. Further in vivo study in the H460 xenograft mouse model validated the anti-tumour activities of 11i with a greater tumour growth inhibition (TGI, 68.3%) than ß-elemene and SAHA (50.1% and 55.9% respectively) at 60 mg/kg ip dosing, without obvious body weight loss and toxicity. Thus, such N-alkyl-N-hydroxyl carboximate class of compounds exemplified as 11i demonstrated potent anticancer activities both in vitro and in vivo, and should warrant further investigation for potential anticancer therapy.

Recent Progress in Fragmentation of Katritzky Salts Enabling Formation of C-C, C-B, and C-S Bonds.

Since their discovery in 1970s, Katritzky salts have emerged as one of the most important classes of building blocks for use in organic synthesis and drug discovery. These bulky pyridinium salts derived from alkylamine can readily generate alkyl radical and undergo a variety of organic transformation reactions such as alkylation, arylation, alkenylation, alkynylation, carbonylation, sulfonylation, and borylation. Through these transformations, complexed molecules bearing new C-C, C-B, or C-S bonds can be constructed in easy ways and in simple steps. This review aims to summarize recent advances in these versatile building blocks in well-classified categories. Representative examples and their reaction mechanisms are discussed. The hope is to provide the scientific community with convenient access to collective information and accelerate further research.

Recent Advances in Synergistic Antitumor Effects Exploited from the Inhibition of Ataxia Telangiectasia and RAD3-Related Protein Kinase (ATR).

As one of the key phosphatidylinositol 3-kinase-related kinases (PIKKs) family members, ataxia telangiectasia and RAD3-related protein kinase (ATR) is crucial in maintaining mammalian cell genomic integrity in DNA damage response (DDR) and repair pathways. Dysregulation of ATR has been found across different cancer types. In recent years, the inhibition of ATR has been proven to be effective in cancer therapy in preclinical and clinical studies. Importantly, tumor-specific alterations such as ATM loss and Cyclin E1 (CCNE1) amplification are more sensitive to ATR inhibition and are being exploited in synthetic lethality (SL) strategy. Besides SL, synergistic anticancer effects involving ATRi have been reported in an increasing number in recent years. This review focuses on the recent advances in different forms of synergistic antitumor effects, summarizes the pharmacological benefits and ongoing clinical trials behind the biological mechanism, and provides perspectives for future challenges and opportunities. The hope is to draw awareness to the community that targeting ATR should have great potential in developing effective anticancer medicines.

Hounsfield Unit Values in ACR TI-RADS 4-5 Thyroid Nodules with Coarse Calcifications: An Important Imaging Feature Helpful for Diagnosis.

PURPOSE: The purpose of this study is to investigate the diagnostic role of Hounsfield unit (HU) values on noncontrast computed tomography (CT) for differentiating benignity from malignancy in the American College of Radiology (ACR) Thyroid Imaging Reporting and Data System (TI-RADS) 4-5 nodules with coarse calcifications. PATIENTS AND METHODS: CT images of 216 ACR TI-RADS 4-5 nodules with coarse calcifications from 207 patients who underwent surgery in our hospital between 2017 and 2019 were retrospectively reviewed. The average HU values (AHUVs) and maximum HU values (MHUVs) of the nodules were measured on noncontrast CT. The distribution of AHUVs and MHUVs in benign and malignant nodules with coarse calcifications was analyzed using the Mann-Whitney test. Receiver operating characteristic (ROC) curves were used to identify the best cut-off values. Diagnostic performances were assessed according to the area under the ROC curve (AUC), sensitivity and specificity. RESULTS: Of the 216 ACR TI-RADS 4-5 nodules with coarse calcifications, 170 were benign and 46 were malignant. The AHUVs of benign and malignant nodules were 791 HU [interquartile range (IQR), 543-1025 HU] and 486 HU (IQR, 406-670 HU), respectively (P < 0.001). The MHUVs of benign and malignant nodules were 1084 HU (IQR, 717-1477 HU) and 677 HU (IQR, 441-986 HU), respectively (P < 0.001). The AUCs for AHUVs and MHUVs for predicting benign nodules with coarse calcifications were 0.759 and 0.732, and the cut-off values were 627.5 HU and 806.0 HU, with sensitivities of 67.6% and 68.8% and specificities of 73.9% and 67.4%, respectively. The sensitivity and specificity of the combination were 68.8% and 76.1%. CONCLUSION: AHUVs and MHUVs were helpful in differentiating benignity from malignancy in ACR TI-RADS 4-5 nodules with coarse calcifications. This may provide an important basis for reducing misdiagnosis and unnecessary aspiration or surgical trauma.

Optimization of Protein-Protein Interaction Measurements for Drug Discovery Using AFM Force Spectroscopy.

Increasingly targeted in drug discovery, protein-protein interactions challenge current high throughput screening technologies in the pharmaceutical industry. Developing an effective and efficient method for screening small molecules or compounds is critical to accelerate the discovery of ligands for enzymes, receptors and other pharmaceutical targets. Here, we report developments of methods to increase the signal-to-noise ratio (SNR) for screening protein-protein interactions using atomic force microscopy (AFM) force spectroscopy. We have demonstrated the effectiveness of these developments on detecting the binding process between focal adhesion kinases (FAK) with protein kinase B (Akt1), which is a target for potential cancer drugs. These developments include optimized probe and substrate functionalization processes and redesigned probe-substrate contact regimes. Furthermore, a statistical-based data processing method was developed to enhance the contrast of the experimental data. Collectively, these results demonstrate the potential of the AFM force spectroscopy in automating drug screening with high throughput.

Regulatory effect of NOX-4 on PI3K signaling pathway in TGF-β1-in-duced collagen Ⅰ synthesis from lung cancer cells / 中国病理生理杂志

AIM:To investigate the regulatory effect of NADPH oxidase-4 (NOX-4) on PI3K signaling path-way in transforming growth factor-β1 (TGF-β1)-induced collagen type Ⅰ (collagen Ⅰ) synthesis from lung cancer cells and the mechanisms. METHODS:Human lung cancer A549 cells were cultured in vitro and stimulated with TGF-β1. The ex-pression of NOX family and collagen family at mRNA and protein levels as well as the PI3K class Ⅰ catalytic subunits and the activation of PI3K signaling pathway was measured. A549 cells were pre-treated with NOX-4 inhibitor diphenyleneiodo-nium (DPI), and the expression of collagen Ⅰ at mRNA level as well as the PI3K class Ⅰ catalytic subunits and the activa-tion of PI3K signaling pathway was measured upon TGF-β1 stimulation. RESULTS:TGF-β1 stimulated the expression of NOX-4 and collagen Ⅰ at mRNA and protein levels as well as the expression of PIK3CD and the activation of PI3K signaling pathway at a dose- and time-dependent manner. NOX-4 inhibitor DPI partly reversed TGF-β1-induced collagen Ⅰ expres-sion. Inhibition of NOX-4 down-regulated the degree of TGF-β1-stimulated activation of PI3K signaling pathway without effect on the expression of PIK3CD. CONCLUSION:NOX-4 participates in TGF-β1-induced collagen Ⅰ synthesis from lung cancer cells via regulating the activation of PI3K signaling pathway. TGF-β1/NOX-4/PI3K signaling pathway axis acts as a regulatory role in collagen Ⅰ synthesis from lung cancer cells.

Progress on role of CCN1 in pulmonary diseases / 中国病理生理杂志

Cysteine-rich angiogenic inducer 61(Cyr61/CCN1)is an extracellular matrix-associated signaling protein consisting of 381 amino-acid residues ,which has the regulatory function for a multitude of cellular responses.The pleiotropic effects of CCN 1 on the initiation and resolution of inflammation as well as oncogenesis and development of tumor were reported.According to the numerous data from experimental and clinical studies ,this article provides an overview on CCN1 and summarizes the latest understanding of the role of CCN 1 in pulmonary diseases.

The biological research brogress in evaluation of lymph Node Metastasis in bladder cancer / 医学研究生学报

Invasion and metastasis are the main behavior of malignant tumors. Lymph node metastasis is an early metastasis mode of bladder cancer, which is an important risk factor for poor prognosis . It is also an important index to decide the treatment, especially to determine the operation mode and strategy. Therefore, searching for the evaluation index of lymph node metastasis of malignant tumor is the focus of tumor research. In recent years, the literature of molecular biology for predicting the staging of bladder cancer has been reviewed, but the literature on biological assessment of lymph node metastasis hasn′t. At present, many evaluation indexes related to lymph node metastasis of bladder cancer have been reported in addition to the new imaging Methods , from gene to protein, and then to cell and tissue. In this paper,we try to look into the biological research progress in the evaluation of lymph node metastasis of bladder cancer.

Calcifying Pseudoneoplasm of the Neuraxis Arising From the Sacral Canal: Case Report and Review of the Literature.

BACKGROUND: Calcifying pseudoneoplasm of the spine is an extremely rare lesion and has not yet been reported in the sacral canal. CASE DESCRIPTION: We present the first known case of a calcifying pseudoneoplasm of the neuraxis arising from the sacral canal and discuss the clinicopathological features and neuroradiological findings of this lesion. CONCLUSIONS: Calcifying pseudoneoplasm of the neuraxis can occur throughout the spine, including the sacral canal, and should be included in the differential diagnosis of intraspinal calcified lesions.

Assessment of pressure-volume loop in patients with pulmonary hypertension and the correlation with New York Heart Function Classification / 中国介入心脏病学杂志

Objective To analyze the correlation between the pressure volume parameters and cardiac function in terms of New York Heart Association(NYHA) classification in patients with pulmonary arterial hypertension. Methods Among 36 patients with pulmonary hypertension admitted in our center between April 2015 to June 2016, right heart catheterization recording right atrial pressure curve,right ventricular pressure curve,pulmonary arterial pressure and oxygen saturation curve in different parts was performal.All patients recived underwent cardiac MRI examination to obtain a single cardiac cycle and the use of Mass software to measure right ventricular volume continuously and right ventricular pressure-volume loop parameters were then obtained. Patients were divided into different groups according to the NYHA functional classification, and the correlation between the parameters of each group and the cardiac function classes a were analyzed. Results Compare to patients with NYHA class Ⅰheart function,patients with NYHA heart function class Ⅱ and class Ⅲ had significantly higher right ventricular end diastolic pressure(P < 0.05)and higher right ventricular systolic pressure (P < 0.01). Spearman correlation analysis showed that cardiac function of NYHA classⅠ,Ⅱ and Ⅲhad position correlation with RVESV,RVEDP and RVESP wheras negative correlation with RVEDV. ROC curve analysis showed that,when the patients were divided into 2 groups as NYHAⅠvs. NYHA class Ⅱ + Ⅲ,NYHA classification for predicting the outcome of the NYHA class Ⅱ + Ⅲ level, pulmonary artery elasticity and right ventricular end systolic pressure had larger area under curves respectively. Conclusions Pressure-volume parameters of right ventricles are more objective indicators for cardiac function assessment for pulmonary hypertension patients and evaluation of disease progression especialy in patients with mild symptoms.

Assessment of pressure-volume loop in patients with pulmonary hypertension and the correlation with New York Heart Function Classification / 中国介入心脏病学杂志

Objective To analyze the correlation between the pressure volume parameters and cardiac function in terms of New York Heart Association(NYHA) classification in patients with pulmonary arterial hypertension. Methods Among 36 patients with pulmonary hypertension admitted in our center between April 2015 to June 2016, right heart catheterization recording right atrial pressure curve,right ventricular pressure curve,pulmonary arterial pressure and oxygen saturation curve in different parts was performal.All patients recived underwent cardiac MRI examination to obtain a single cardiac cycle and the use of Mass software to measure right ventricular volume continuously and right ventricular pressure-volume loop parameters were then obtained. Patients were divided into different groups according to the NYHA functional classification, and the correlation between the parameters of each group and the cardiac function classes a were analyzed. Results Compare to patients with NYHA class Ⅰheart function,patients with NYHA heart function class Ⅱ and class Ⅲ had significantly higher right ventricular end diastolic pressure(P < 0.05)and higher right ventricular systolic pressure (P < 0.01). Spearman correlation analysis showed that cardiac function of NYHA classⅠ,Ⅱ and Ⅲhad position correlation with RVESV,RVEDP and RVESP wheras negative correlation with RVEDV. ROC curve analysis showed that,when the patients were divided into 2 groups as NYHAⅠvs. NYHA class Ⅱ + Ⅲ,NYHA classification for predicting the outcome of the NYHA class Ⅱ + Ⅲ level, pulmonary artery elasticity and right ventricular end systolic pressure had larger area under curves respectively. Conclusions Pressure-volume parameters of right ventricles are more objective indicators for cardiac function assessment for pulmonary hypertension patients and evaluation of disease progression especialy in patients with mild symptoms.

Deferoxamine released from poly(lactic-co-glycolic acid) promotes healing of osteoporotic bone defect via enhanced angiogenesis and osteogenesis.

The regeneration capacity of osteoporotic bones is generally lower than that of normal bones. Current methods of osteoporotic bone defect treatment are not always satisfactory. Recent studies demonstrate that activation of the hypoxia inducible factor-1α (HIF-1α) pathway, by genetic methods or hypoxia-mimicking agents, could accelerate bone regeneration. However, little is known as to whether modulating the HIF-1α pathway promotes osteoporotic defect healing. To address this problem in the present study, we first demonstrated that HIF-1α and vascular endothelial growth factor expression levels are lower in osteoporotic bones than in normal bones. Second, we loaded poly(Lactic-co-glycolic acid) (PLGA) with the hypoxia-mimetic agent deferoxamine (DFO). DFO released from PLGA had no significant effect on the proliferation of mesenchymal stem cells (MSCs); however, DFO did enhance the osteogenic differentiation of MSCs. In addition, DFO upregulated the mRNA expression levels of angiogenic factors in MSCs. Endothelial tubule formation assays demonstrate that DFO promoted angiogenesis in human umbilical vein endothelial cells. Third, untreated PLGA scaffolds (PLGA group) or DFO-containing PLGA (PLGA + DFO group) were implanted into critically sized osteoporotic femur defects in ovariectomized rats. After treatment periods of 14 or 28 days, micro-CT, histological, CD31 immunohistochemical, and dynamic bone histomorphometric analyses showed that DFO dramatically stimulated bone formation and angiogenesis in a critically sized osteoporotic femur defect model. Our in vitro and in vivo results demonstrate that DFO may promote the healing of osteoporotic bone defects due to enhanced angiogenesis and osteogenesis. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2515-2527, 2016.

Metastatic Adenocarcinoma of Unknown Primary in the Bilateral Cerebellopontine Angles: Case Report and Review of the Literature.

Metastatic adenocarcinoma in bilateral cerebellopontine angles (CPA) is rare. We report a case and review the current literature in order to enhance recognition of metastatic adenocarcinoma in the cerebellopontine angle. A 44-year-old man was referred to the hospital with rightsided diminished hearing for 7 weeks, left-sided facial palsy for 2 weeks, and left-sided sensorineural hearing loss for 1 week. On Magnetic Resonance Imaging (MRI) two tumors in bilateral CPAs were detected. The left-sided tumor was resected and histopathological examination revealed an adenocarcinoma. Many investigations could not find the primary tumor. One should be careful with middle-aged or elderly patients with sudden progressive deficits in the VIII < sup > th < /sup > or VII < sup > th < /sup > cranial nerves, particularly in bilateral CPA.

Activation of HIFa pathway in mature osteoblasts disrupts the integrity of the osteocyte/canalicular network.

The hypoxia-inducible factors (HIFs), HIF-1α and HIF-2α, are the central mediators of the homeostatic response that enables cells to survive and differentiate in low-oxygen conditions. Previous studies indicated that disruption of the von Hippel-Lindau gene (Vhl) coincides with the activation of HIFα signaling. Here we show that inactivation of Vhl in mature osteoblasts/osteocytes induces their apoptosis and disrupts the cell/canalicular network. VHL-deficient (ΔVHL) mice exhibited a significantly increased cortical bone area resulting from enhanced proliferation and osteogenic differentiation of the bone marrow stromal cells (BMSCs) by inducing the expression of ß-catenin in the BMSC. Our data suggest that the VHL/HIFα pathway in mature osteoblasts/osteocytes plays a critical role in the bone cell/canalicular network and that the changes of osteocyte morphology/function and cell/canalicular network may unleash the bone formation, The underlying mechanism of which was the accumulation of ß-catenin in the osteoblasts/osteoprogenitors of the bone marrow.