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PKCαinhibitors promote breast cancer immune evasion by maintaining PD-L1 stability

Acta Pharmaceutica Sinica B 2024年第10期14卷 4378-4395页

作者： Jiaojiao Yu Yujin Xiang Yuzhen Gao Shan Chang Ren Kong Xiaoxi Lv Jinmei Yu Yunjie Jin Chenxi Li Yiran Ma Zhenhe Wang Jichao Zhou Hongyu Yuan Shuang Shang Fang Hua Xiaowei Zhang Bing Cui Pingping Li State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China CAMS Key Laboratory of Molecular Mechanism and Target Discovery of Metabolic Disorder and Tumorigenesis Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Institute of Bioinformatics and Medical Engineering School of Electrical and Information EngineeringJiangsu University of TechnologyChangzhou 213001China

Protein kinase Cα(PKCα)regulates diverse biological functions of cancer cells and is a promising therapeutic ***,clinical trials of PKC-targeted therapies have not yielded satisfactory *** studies have also indicated a tumor-suppressive role of PKCs via unclear molecular *** this study,we found that PKCαinhibition enhances CD8+T-cell-mediated tumor evasion and abolishes antitumor activity in immunocompetent *** further identified PKCαas a critical regulator of programmed cell death-ligand 1(PD-L1)and found that it enhances T-cell-dependent antitumor immunity in breast cancer by interacting with PD-L1 and suppressing PD-L1 *** demonstrated that PKCα-mediated PD-L1 phosphorylation promotes PD-L1 degradation throughβtransducin repeat-containing ***,the efficacy of PKCαinhibitors was intensified by synergizing with anti-PD-L1 mAb therapy to boost antitumor T-cell immunity in *** analysis revealed that PKCαexpression is positively correlated with T-cell function and the interferon-gamma signature in patients with breast *** study demonstrated the antitumor capability of PKCα,identified potential therapeutic strategies to avoid tumor evasion via PKC-targeted therapies,and provided a proof of concept for targeting PKCαin combination with anti-PD-L1 mAb therapy as a potential therapeutic approach against breast cancer,especially TNBC.

关键词： Protein kinase C PD-L1 β-TRCP Degradation Immune evasion Immunotherapy Combination strategies Breast cancer

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Spatial metabolomics highlights metabolic reprogramming in acute myeloid leukemia mice through creatine pathway

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Acta Pharmaceutica Sinica B 2024年第10期14卷 4461-4477页

作者： Yucheng Bao Jing Qiao Wenjie Gong Ruihong Zhang Yanting Zhou Yinyin Xie Yuan Xie Jiuming He Tong Yin Shanghai Institute of Hematology State Key Laboratory of Medical GenomicsNational Research Centre for Translational Medicine at ShanghaiRuijin Hospital affiliated to Shanghai Jiao Tong University School of MedicineShanghai 200025China College of Osteopathic Medicine Kansas City UniversityKansas CityMO 64106USA State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China NMPA Key Laboratory for Safety Research and Evaluation of Innovative Drug Beijing 100050China

Acute myeloid leukemia(AML)is recognized as an aggressive cancer that is characterized by significant metabolic ***,we applied spatial metabolomics to achieve high-throughput,in situ identification of metabolites within the liver metastases of AML *** at metabolite and protein levels were further mapped out and validated by integrating untargeted metabolomics and *** study showed a downregulation in arginine's contribution to polyamine biosynthesis and urea cycle,coupled with an upregulation of the creatine *** upregulation of creatine synthetases Gatm and Gamt,as well as the creatine transporter Slc6a8,resulted in a marked accumulation of creatine within tumor *** process further enhances oxidative phosphorylation and glycolysis of leukemia cells,thereby boosting ATP production to foster proliferation and ***,we discovered that inhibiting Slc6a8 can counter these detrimental effects,offering a new strategy for treating AML by targeting metabolic pathways.

关键词： Spatial metabolomics Acute myeloid leukemia Metabolic reprogramming Creatine Slc6a8 Oxidative phosphorylation Glycolysis Metastasis

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Pharmacometabolomics and mass spectrometry imaging approach to reveal the neurochemical mechanisms of Polygala tenuifolia

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Journal of Pharmaceutical Analysis 2024年第7期14卷 1035-1046页

作者： Qian Li Jinpeng Bai Yuxue Ma Yu Sun Wenbin Zhou Zhaoying Wang Zhi Zhou Zhonghua Wang Yanhua Chen Zeper Abliz Key Laboratory of Mass Spectrometry Imaging and Metabolomics(National Ethnic Affairs Commission) Minzu University of ChinaBeijing100081China Key Laboratory of Ethnomedicine of Ministry of Education School of PharmacyMinzu University of ChinaBeijing100081China Center for Imaging and Systems Biology College of Life and Environmental SciencesMinzu University of ChinaBeijing100081China State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical SciencesPeking Union Medical CollegeBeijing100050China

Polygala tenuifolia,commonly known as Yuanzhi(YZ)in Chinese,has been shown to possess antiinsomnia ***,the material basis and the mechanism underlying its sedative-hypnotic effects remain ***,we investigated the active components and neurochemical mechanism of YZ extracts using liquid chromatography tandem mass spectrometry(LC-MS/MS)-based pharmacometabolomics and mass spectrometry imaging(MSI)-based spatial resolved *** to the results,17 prototypes out of 101 ingredients in the YZ extract were detected in both the plasma and brain,which might be the major components contributing to the sedative-hypnotic *** pharmacology analysis revealed that these prototypes may exert their effects through neuroactive ligand-receptor interaction,serotonergic synapse,dopaminergic synapse,and dopaminergic synapse,among other ***-MS/MS-based targeted metabolomics and Western blot(WB)revealed that tryptophan-serotonin-melatonin(Trp-5-HT-Mel)and tyrosine-norepinephrine-adrenaline(Tyr-Ne-Ad)are the key regulated *** decarboxylase(DDC)upregulation and phenylethanolamine Nmethyltransferase(PNMT)downregulation further confirmed these ***,MSI-based spatially resolved metabolomics revealed notable alterations in 5-HT in the pineal gland(PG),and Ad in the brainstem,including the middle brain(MB),pons(PN),and hypothalamus(HY).In summary,this study illustrates the efficacy of an integrated multidimensional metabolomics approach in unraveling the sedative-hypnotic effects and neurochemical mechanisms of a Chinese herbal medicine,YZ.

关键词： Polygala tenuifolia Pharmaceutical metabolomics Spatially resolved metabolome Sedative-hypnotic Neurochemical mechanisms

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Spatiotemporally resolved metabolomics and isotope tracing reveal CNS drug targets

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Acta Pharmaceutica Sinica B 2023年第4期13卷 1699-1710页

作者： Bo Jin Xuechao Pang Qingce Zang Man Ga Jing Xu Zhigang Luo Ruiping Zhang Jiangong Shi Jiuming He Zeper Abliz State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China NMPA Key Laboratory for Safety Research and Evaluation of Innovative Drug Beijing 100050China Key Laboratory of Mass Spectrometry Imaging and Metabolomics(Minzu University of China) National Ethnic Affairs CommissionBeijing 100081China Key Laboratory of Ethnomedicine of Ministry of Education School of PharmacyMinzu University of ChinaBeijing 100081China

Deconvolution of potential drug targets of the central nervous system(CNS)is particularly challenging because of the complicated structure and function of the ***,a spatiotemporally resolved metabolomics and isotope tracing strategy was proposed and demonstrated to be powerful for deconvoluting and localizing potential targets of CNS drugs by using ambient mass spectrometry *** strategy can map various substances including exogenous drugs,isotopically labeled metabolites,and various types of endogenous metabolites in the brain tissue sections to illustrate their microregional distribution pattern in the brain and locate drug action-related metabolic nodes and *** strategy revealed that the sedative-hypnotic drug candidate YZG-331 was prominently distributed in the pineal gland and entered the thalamus and hypothalamus in relatively small amounts,and can increase glutamate decarboxylase activity to elevateγ-aminobutyric acid(GABA)levels in the hypothalamus,agonize organic cation transporter 3 to release extracellular histamine into peripheral *** findings emphasize the promising capability of spatiotemporally resolved metabolomics and isotope tracing to help elucidate the multiple targets and the mechanisms of action of CNS drugs.

关键词： Ambient mass spectrometry imaging Spatiotemporally resolved metabolomics Isotope tracing Drug targets Central nervous system Drug candidate

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Pyk2 inhibition attenuates hypoxic-ischemic brain injury in neonatal mice

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中国药理学报（英文版） 2022年第4期43卷 797-810页

作者： Jie Zhu Shi-feng Chu Ye Peng Dan-dan Liu Chen Chen Wen-xuan Jian Hong-shuo Sun Zhong-ping Feng Zhao Zhang Nai-hong Chen State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia Medica&Neuroscience CenterChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia Medica&Neuroscience CenterChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China College of Pharmacy Hunan University of Chinese MedicineChangsha 410208China State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia Medica&Neuroscience CenterChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Institute of Clinical Pharmacology Guangzhou University of Chinese MedicineGuangzhou 510720China Department of Physiology Faculty of MedicineUniversity of TorontoTorontoONCanada State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia Medica&Neuroscience CenterChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China College of Pharmacy Hunan University of Chinese MedicineChangsha 410208China Institute of Clinical Pharmacology Guangzhou University of Chinese MedicineGuangzhou 510720China

Newborns suffering from hypoxia-ischemia(HI)brain injury still lack effective ***-rich tyrosine kinase 2(Pyk2)is a non-receptor tyrosine kinase,which is highly correlated with transient ischemic brain injury in *** this study,we investigated the role of Pyk2 in neonatal HI brain *** was induced in postnatal day 7 mouse pups by unilateral common carotid artery ligation followed by hypoxic ***2 interference lentivirus(LV-Pyk2 shRNA)was constructed and injected into unilateral cerebral ventricle of neonatal mice before *** volume,pathological changes,and neurological behaviors were assessed on postnatal day *** showed that the phosphorylation level of Pyk2 was significantly increased in neonatal brain after HI,whereas LV-Pyk2 shRNA injection significantly attenuated acute HI brain damage and improved neurobehavioral *** oxygen-glucose deprivation-treated cultured cortical neurons,Pyk2 inhibition significantly alleviated NMDA receptor-mediated excitotoxicity;similar results were also observed in neonatal HI brain *** demonstrated that Pyk2 inhibition contributes to the long-term cerebrovascular recovery assessed by laser speckle contrast imaging,but cognitive function was not obviously improved as evaluated in Morris water maze and novel object recognition ***,we constructed lentiviral LV-HIF-Pyk2 shRNA,through which HIF-1 α promoter-mediated interference of Pyk2 would occur during the anoxic *** injection of LV-HIF-Pyk2 shRNA significantly improved long-term recovery of cognitive function in HI-treated neonatal *** conclusion,this study demonstrates that Pyk2 interference protects neonatal brain from hypoxic-ischemic ***-1 α promoter-mediated hypoxia conditional control is a useful tool to distinguish between hypoxic period and normal ***2 is a promising drug target for potential treatment of neonatal HI brain injury.

关键词： neonatal mice hypoxic-ischemic brain injury Pyk2 NMDA receptor excitotoxicity HIF-1α

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Identification of the cytochrome P450s responsible for the biosynthesis of two types of aporphine alkaloids and their de novo biosynthesis in yeast

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Journal of Integrative Plant Biology 2024年第8期66卷 1703-1717页

作者： Qishuang Li Xiang Jiao Xinyi Li Wenlong Shi Ying Ma Xiangmei Tan Jingyi Gan Jimei Liu Jian Yang Jian Wang Baolong Jin Tong Chen Ping Su Yujun Zhao Yifeng Zhang Jinfu Tang Guanghong Cui Yun Chen Juan Guo Luqi Huang School of Traditional Chinese Pharmacy China Pharmaceutical UniversityNanjing 211198China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs National Resource Center for Chinese Materia MedicaChina Academy of Chinese Medical SciencesBeijing 100700China Department of Life Sciences Chalmers University of TechnologyGothenburg SE-41296Sweden State Key Laboratory of Natural and Biomimetic Drugs School of Pharmaceutical SciencesPeking UniversityBeijing 100191China State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaThe Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China

Aporphine alkaloids have diverse pharmacological activities;however,our understanding of their biosynthesis is relatively *** studies have classified aporphine alkaloids into two categories based on the configuration and number of substituents of the D-ring and have proposed preliminary biosynthetic pathways for each *** this study,we identified two specific cytochrome P450 enzymes(CYP80G6 and CYP80Q5)with distinct activities toward(S)-configured and(R)-configured substrates from the herbaceous perennial vine Stephania tetrandra,shedding light on the biosynthetic mechanisms and stereochemical features of these two aporphine alkaloid ***,we characterized two CYP719C enzymes(CYP719C3 and CYP719C4)that catalyzed the formation of the methylenedioxy bridge,an essential pharmacophoric group,on the A-and D-rings,respectively,of aporphine *** the functional characterization of these crucial cytochrome P450 enzymes,we reconstructed the biosynthetic pathways for the two types of aporphine alkaloids in budding yeast(Saccharomyces cerevisiae)for the de novo production of compounds such as(R)-glaziovine,(S)-glaziovine,and *** study provides key insight into the biosynthesis of aporphine alkaloids and lays a foundation for producing these valuable compounds through synthetic biology.

关键词： aporphine alkaloids biosynthesis CYP719C CYP80 engineered yeast

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Baicalein attenuates rotenone-induced SH-SY5Y cell apoptosis through binding to SUR1 and activating ATP-sensitive potassium channels

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中国药理学报（英文版） 2024年第3期45卷 480-489页

作者： De-wen Kong Li-da Du Run-zhe Liu Tian-yi Yuan Shou-bao Wang Yue-hua Wang Yang Lu Lian-hua Fang Guan-hua Du State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Beijing Key Laboratory of Drug Targets Identification and Drug Screening National Center for Pharmaceutical ScreeningInstitute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Shandong Soteria Pharmaceutical Co Ltd. Jinan 250022China State Key Laboratory of Bioactive Substances and Functions of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Beijing Key Laboratory of Drug Targets Identification and Drug Screening National Center for Pharmaceutical ScreeningInstitute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Beijing Key Laboratory of Polymorphic Drugs Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China

Dopaminergic neurons in the substantia nigra(SN)expressing SUR1/Kir6.2 type ATP-sensitive potassium channels(K-ATP)are more vulnerable to rotenone or metabolic stress,which may be an important reason for the selective degeneration of neurons in Parkinson's disease(PD).Baicalein has shown neuroprotective effects in PD animal *** this study,we investigated the effect of baicalein on K-ATP channels and the underlying mechanisms in rotenone-induced apoptosis of SH-SY5Y cells.K-ATP currents were recorded from SH-SY5Y cells using whole-cell voltage-clamp *** dissolved in the external solution at the final concentration were directly pipetted onto the *** showed that rotenone and baicalein opened K-ATP channels and increased the current amplitudes with EC50 values of 0.438 μM and 6.159 μM,respectively.K-ATP channel blockers glibenclamide(50 μM)or 5-hydroxydecanoate(5-HD,250 μM)attenuated the protective effects of baicalein in reducing reactive oxygen species(ROS)content and increasing mitochondrial membrane potential and ATP levels in rotenone-injured SH-SY5Y cells,suggesting that baicalein protected against the apoptosis of SH-SY5Y cells by regulating the effect of rotenone on opening K-ATP *** of baicalein(150,300 mg·kg-1·d-1,i.g.)significantly inhibited rotenone-induced overexpression of SUR1 in SN and striatum of *** conducted surface plasmon resonance assay and molecular docking,and found that baicalein had a higher affinity with SUR1 protein(KD=10.39μM)than glibenclamide(KD=24.32 μM),thus reducing the sensitivity of K-ATP channels to *** of SUR1 subunit reduced rotenone-induced apoptosis and damage of SH-SY5Y cells,confirming that SURI was an important target for slowing dopaminergic neuronal degeneration in *** together,we demonstrate for the first time that baicalein attenuates rotenone-induced SH-SY5Y cell apoptosis through binding to SUR1 and activating K-ATP channels.

关键词： Parkinson’s disease baicalein SH-SY5Y cells K-ATP channels SUR1 subunit rotenone

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Design and characterizing of robust probes for enhanced mass spectrometry imaging and spatially resolved metabolomics

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Chinese Chemical Letters 2023年第5期34卷 397-402页

作者： Jianpeng Huang Shanshan Gao Kai Wang Jin Zhang Xuechao Pang Junwen Shi Jiuming He State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China NMPA Key Laboratory of Safety Research and Evaluation of Innovative Drug Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Viktor Technology Co. Ltd.Beijing 101100China

As for the emerging and cut edge spatially resolved metabolomics,mass spectrometry imaging(MSI)is a powerful tool that can map thousands of metabolites from bio-tissue sections without chemical ***,the stability,sensitivity and spatial resolution of MSI are always limited by the performance of its ionization ***,two types of probes(fine probe(P-100)and large probe(P-200))were designed and characterized to perform air-flow assisted desorption electrospray ionization(AFA-DESI)MSI analysis for spatially resolved *** was determined that the spray introduced by P-100 was homogenous and stable under the spray solvent at a flow rate of 5-10μL/min,while P-200 can endure a high flow rate of up to 10-30μL/***,the MSI images were acquired by AFA-DESI-MSI with P-100 from rat brain tissue section and with P-200 from whole-body tissue section of mouse,and these results presented unambiguous tissue structure with the distribution information of numerous ***,the spatially resolved metabolomic analysis of tumor tissue was successfully realized to discover the tumor associated *** the key parts of AFA-DESI-MSI system,it has been demonstrated that the designed probs have excellent performance for spatially resolved metabolomics,and it will further promote its application in life science,and drug research and development.

关键词： Air-flow assisted desorption electrospray ionization Mass spectrometry imaging Molecular imaging Spatially resolved metabolomics Brain

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Bone-derived MSCs encapsulated in alginate hydrogel prevent collagen-induced arthritis in mice through the activation of adenosine A_(2A/2B)receptors in tolerogenic dendritic cells

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Acta Pharmaceutica Sinica B 2023年第6期13卷 2778-2794页

作者： Gaona Shi Yu Zhou Wenshuai Liu Chengjuan Chen Yazi Wei Xinlong Yan Lei Wu Weiwei Wang Lan Sun Tiantai Zhang State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Tianjin Key Laboratory of Biomaterial Research Institute of Biomedical EngineeringChinese Academy of Medical Sciences&Peking Union Medical CollegeTianjin 300192China Faculty of Environmental and Life Sciences Beijing University of TechnologyBeijing 100124China

Tolerogenic dendritic cells(tol DCs)facilitate the suppression of autoimmune responses by differentiating regulatory T cells(Treg).The dysfunction of immunotolerance results in the development of autoimmune diseases,such as rheumatoid arthritis(RA).As multipotent progenitor cells,mesenchymal stem cells(MSCs),can regulate dendritic cells(DCs)to restore their immunosuppressive function and prevent disease ***,the underlying mechanisms of MSCs in regulating DCs still need to be better ***,the delivery system for MSCs also influences their ***,MSCs are encapsulated in alginate hydrogel to improve cell survival and retention in situ,maximizing efficacy in *** three-dimensional co-culture of encapsulated MSCs with DCs demonstrates that MSCs can inhibit the maturation of DCs and the secretion of pro-inflammatory *** the collagen-induced arthritis(CIA)mice model,alginate hydrogel encapsulated MSCs induce a significantly higher expression of CD39^(+)CD73^(+)on *** enzymes hydrolyze ATP to adenosine and activate A_(2A/2B)receptors on immature DCs,further promoting the phenotypic transformation of DCs to tol DCs and regulating naive T cells to ***,encapsulated MSCs obviously alleviate the inflammatory response and prevent CIA *** finding clarifies the mechanism of MSCs-DCs crosstalk in eliciting the immunosuppression effect and provides insights into hydrogel-promoted stem cell therapy for autoimmune diseases.

关键词： Mesenchymal stem cells Alginate hydrogel Tolerogenic dendritic cells Immunotolerance Adenosine A_(2A/2B)receptor CD39/CD73 Treg Rheumatoid arthritis

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Spatiotemporal pharmacometabolomics based on ambient mass spectrometry imaging to evaluate the metabolism and hepatotoxicity of amiodarone in HepG2 spheroids

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Journal of Pharmaceutical Analysis 2023年第5期13卷 483-493页

作者： Limei Li Qingce Zang Xinzhu Li Ying Zhu Shanjing Wen Jiuming He Ruiping Zhang Zeper Abliz State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing100050China Key Laboratory of Mass Spectrometry Imaging and Metabolomics(Minzu University of China) National Ethnic Affairs CommissionBeijing100081China Center for Imaging and Systems Biology College of Life and Environmental SciencesMinzu University of ChinaBeijing100081China

Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and pathological ***,airflow-assisted desorption electrospray ionization-MSI(AFADESI-MSI)was coupled with 3D HepG2 spheroids to assess the metabolism and hepatotoxicity of amiodarone(AMI).High-coverage imaging of>1100 endogenous metabolites in hepatocyte spheroids was achieved using *** AMI treatment at different times,15 metabolites of AMI involved in Ndesethylation,hydroxylation,deiodination,and desaturation metabolic reactions were identified,and according to their spatiotemporal dynamics features,the metabolic pathways of AMI were ***,the temporal and spatial changes in metabolic disturbance within spheroids caused by drug exposure were obtained via metabolomic *** main dysregulated metabolic pathways included arachidonic acid and glycerophospholipid metabolism,providing considerable evidence for the mechanism of AMI *** addition,a biomarker group of eight fatty acids was selected that provided improved indication of cell viability and could characterize the hepatotoxicity of *** combination of AFADESI-MSI and HepG2 spheroids can simultaneously obtain spatiotemporal information for drugs,drug metabolites,and endogenous metabolites after AMI treatment,providing an effective tool for in vitro drug hepatotoxicity evaluation.

关键词： Mass spectrometry imaging HepG2 spheroids Hepatotoxicity Drug metabolism Amiodarone

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