[go: up one dir, main page]

CN118845751A - Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection - Google Patents

Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection Download PDF

Info

Publication number
CN118845751A
CN118845751A CN202410829865.2A CN202410829865A CN118845751A CN 118845751 A CN118845751 A CN 118845751A CN 202410829865 A CN202410829865 A CN 202410829865A CN 118845751 A CN118845751 A CN 118845751A
Authority
CN
China
Prior art keywords
ifn
kaempferol
virus
type
metabolites
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202410829865.2A
Other languages
Chinese (zh)
Inventor
杜瑞坤
宁云佳
崔清华
荣立军
陈子诺
陈成龙
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shandong University of Traditional Chinese Medicine
Original Assignee
Shandong University of Traditional Chinese Medicine
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shandong University of Traditional Chinese Medicine filed Critical Shandong University of Traditional Chinese Medicine
Priority to CN202410829865.2A priority Critical patent/CN118845751A/en
Publication of CN118845751A publication Critical patent/CN118845751A/en
Pending legal-status Critical Current

Links

Classifications

    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

本发明涉及山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的应用,所述病毒包括甲型流感病毒、水泡性口腔炎病毒、严重发热伴血小板减少综合征病毒或克里米亚‑刚果出血热病毒中的任意一种或至少两种的组合。本发明证明了山奈素及其代谢产物能够增强和延长I型IFN激活的JAK/STAT通路,增强固有免疫,具有广谱抗病毒活性。

The present invention relates to the use of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection, wherein the viruses include any one or a combination of at least two of influenza A virus, vesicular stomatitis virus, severe fever with thrombocytopenia syndrome virus or Crimean-Congo hemorrhagic fever virus. The present invention proves that kaempferol and its metabolites can enhance and prolong the JAK/STAT pathway activated by type I IFN, enhance innate immunity, and have broad-spectrum antiviral activity.

Description

山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的 应用Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection

技术领域Technical Field

本发明涉及生物医药领域,尤其涉及一种山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的应用。The present invention relates to the field of biomedicine, and in particular to application of kaempferol and/or kaempferol metabolites in the preparation of drugs for resisting viral infection.

背景技术Background Art

宿主抗病毒免疫,特别是先天性抗病毒免疫,是一种通用的防御机制,对所有病原体都具有非特异性作用。在复杂而丰富的抗病毒先天免疫反应中,I型干扰素(IFN-α/β)反应是第一道防线,在宿主抵御病毒入侵方面发挥着核心作用。在病毒感染后,IFN的表达被触发。随后,IFN的分泌可被感染细胞和周围未感染细胞表面的IFN受体(IFNARs)识别,通过Janus激酶信号转导子和转录激活子(JAK/STAT)途径启动快速信号转导,从而形成由磷酸化STAT1、STAT2和IRF9组成的转录复合物。然后,这种转录复合物被转运到细胞核中,并与许多干扰素刺激基因(ISG)启动子区中的特定干扰素刺激反应元件(ISRE)基序结合,其中大多数具有抗病毒特性,从而引发反应细胞的抗病毒状态。Host antiviral immunity, especially innate antiviral immunity, is a universal defense mechanism that has non-specific effects on all pathogens. Among the complex and rich antiviral innate immune responses, the type I interferon (IFN-α/β) response is the first line of defense and plays a central role in the host's defense against viral invasion. After viral infection, IFN expression is triggered. Subsequently, IFN secretion can be recognized by IFN receptors (IFNARs) on the surface of infected cells and surrounding uninfected cells, initiating rapid signal transduction through the Janus kinase signal transducer and activator of transcription (JAK/STAT) pathway, thereby forming a transcription complex composed of phosphorylated STAT1, STAT2 and IRF9. This transcription complex is then transported into the nucleus and binds to specific interferon-stimulated response element (ISRE) motifs in the promoter region of many interferon-stimulated genes (ISGs), most of which have antiviral properties, thereby inducing the antiviral state of the responding cells.

当新病毒首次出现时,对抗这些新出现的病毒感染的可用药物非常少,尽管一些现有的抗病毒药物,如核苷类似物(利巴韦林、阿昔洛韦、法匹拉韦或瑞德西韦)具有广谱抗病毒效力,但当其应用于新出现的病毒时,其治疗效果甚微。因此,迫切需要开发新的广谱抗病毒制剂,以加强我们对未来新出现病原体爆发的准备。When new viruses first appear, there are very few drugs available to fight these emerging viral infections, and although some existing antiviral drugs, such as nucleoside analogs (ribavirin, acyclovir, favipiravir, or remdesivir), have broad-spectrum antiviral efficacy, they have little therapeutic effect when applied to emerging viruses. Therefore, there is an urgent need to develop new broad-spectrum antiviral agents to enhance our preparedness for future outbreaks of emerging pathogens.

CN110403941A公开了一种广谱抗病毒药物或组合物,所述药物或组合物的有效成分为化合物RAF265或其药学上可接受的盐,首次发现RAF265具备广谱高效的抗病毒活性,可高效抑制6科类7种病毒(PRV、HSV-1、PEDV、FMDV、NDV、VSV、IBDV)感染宿主细胞及小鼠,显著减少小鼠体内病毒量,减轻感染症状。且RAF265细胞毒性较低(CC50/IC50>100),对鸡胚与小鼠无毒副作用。CN110403941A discloses a broad-spectrum antiviral drug or composition, wherein the active ingredient of the drug or composition is the compound RAF265 or a pharmaceutically acceptable salt thereof. It is the first time that RAF265 has a broad-spectrum and highly effective antiviral activity, and can effectively inhibit the infection of host cells and mice by 7 viruses of 6 families (PRV, HSV-1, PEDV, FMDV, NDV, VSV, IBDV), significantly reduce the amount of virus in mice, and alleviate infection symptoms. Moreover, RAF265 has low cytotoxicity (CC50/IC50>100) and has no toxic side effects on chicken embryos and mice.

CN115379852A公开了一种抗病毒药物,其包括2’-O-甲基化敏感RNA相关酶或2’-O-甲基化敏感RNA相关酶的编码核酸序列。还提供了包括该2’-O-甲基化敏感RNA相关酶或2’-O-甲基化敏感RNA相关酶的编码核酸序列的制药用途。该2’-O-甲基化敏感RNA相关酶为来自生殖支原体的核糖核酸酶MgR。所述抗病毒药物具有广谱抗病毒效果,并且基本上无细胞毒性,可用于防止病毒对动植物和人的感染。CN115379852A discloses an antiviral drug, which includes a 2'-O-methylation sensitive RNA related enzyme or a 2'-O-methylation sensitive RNA related enzyme encoding nucleic acid sequence. Also provided is a pharmaceutical use of the 2'-O-methylation sensitive RNA related enzyme or a 2'-O-methylation sensitive RNA related enzyme encoding nucleic acid sequence. The 2'-O-methylation sensitive RNA related enzyme is a ribonuclease MgR from Mycoplasma genitalium. The antiviral drug has a broad-spectrum antiviral effect and is substantially non-cytotoxic, and can be used to prevent viral infection of animals, plants and humans.

黄酮类化合物是一大类天然代谢产物,在植物中含量特别丰富,具有多种生物活性,如抗糖尿病、抗炎、抗菌、抗氧化、抗病毒、细胞毒性和降脂活性。迄今为止,多达19种黄酮类化合物已被批准用于处方,另有20种正在进行临床试验。Flavonoids are a large class of natural metabolites that are particularly abundant in plants and have a variety of biological activities, such as anti-diabetic, anti-inflammatory, antibacterial, antioxidant, antiviral, cytotoxic and lipid-lowering activities. To date, as many as 19 flavonoids have been approved for prescription use, and another 20 are undergoing clinical trials.

综上所述,如何开发一种增强宿主固有免疫的药物,已成为目前本领域亟待解决的问题之一。In summary, how to develop a drug that enhances host innate immunity has become one of the urgent problems to be solved in this field.

发明内容Summary of the invention

为解决上述技术问题,本发明提供了山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的应用,山奈素和/或山奈素通过增强和延长I型IFN激活的JAK/STAT通路,增强固有免疫,具有广谱抗病毒活性。In order to solve the above technical problems, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of drugs for anti-viral infection. Kaempferol and/or kaempferol enhance innate immunity by enhancing and prolonging the JAK/STAT pathway activated by type I IFN, and have broad-spectrum antiviral activity.

为达此目的,本发明采用以下技术方案:To achieve this object, the present invention adopts the following technical solutions:

第一方面,本发明提供了山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的应用。In a first aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection.

山奈素和山奈素代谢产物山奈酚的化学结构如图1所示。本发明通过研究证明,山奈素及其代谢产物是一种IFN的应答剂,能够与I型IFNs协同作用,增强固有免疫,从而达到广谱抗病毒的作用。The chemical structures of kaempferol and kaempferol metabolite are shown in Figure 1. The present invention proves through research that kaempferol and its metabolites are IFN responders, which can synergize with type I IFNs to enhance innate immunity, thereby achieving a broad-spectrum antiviral effect.

优选地,所述病毒包括DNA病毒和/或RNA病毒。Preferably, the virus comprises a DNA virus and/or an RNA virus.

本发明提供的山奈素及其代谢产物具有广谱抗病毒作用,因此对病毒的种类不做限定,除了目前已知的病毒以外,还可以应用于目前尚未出现的新生病毒的抗病毒感染作用中。The kaempferol and its metabolites provided by the present invention have a broad-spectrum antiviral effect, and therefore are not limited to the types of viruses. In addition to currently known viruses, they can also be applied to the antiviral infection effect of newly emerged viruses that have not yet appeared.

优选地,所述DNA病毒包括乙肝病毒、疱疹病毒、人乳头瘤病毒或腺病毒中的任意一种或至少两种的组合。Preferably, the DNA virus comprises any one of hepatitis B virus, herpes virus, human papillomavirus or adenovirus, or a combination of at least two thereof.

优选地,所述RNA病毒包括艾滋病病毒、SARS病毒、MERS病毒、埃博拉病毒、流感病毒、副流感病毒、狂犬病毒、腮腺炎病毒、麻疹病毒、呼吸道合胞病毒、艾柯病毒、黄病毒、甲病毒、布尼亚病毒、甲型肝炎病毒、柯萨奇病毒、鼻病毒、肠道病毒、脊髓灰质炎病毒、登革热病毒、轮状病毒、马尔堡病毒或新型冠状病毒中的任意一种或至少两种的组合。Preferably, the RNA virus comprises any one or a combination of at least two of HIV, SARS virus, MERS virus, Ebola virus, influenza virus, parainfluenza virus, rabies virus, mumps virus, measles virus, respiratory syncytial virus, echovirus, flavivirus, alphavirus, bunyavirus, hepatitis A virus, coxsackievirus, rhinovirus, enterovirus, poliovirus, dengue virus, rotavirus, Marburg virus or the new coronavirus.

优选地,所述病毒包括甲型流感病毒(Influenza AVirus,IAV)、水泡性口腔炎病毒(Vesicular Stomatitis Virus,VSV)、严重发热伴血小板减少综合征病毒(SevereFever with Thrombocytopenia Syndrome Virus,SFTSV)或克里米亚-刚果出血热病毒(Crimean-Congo Hemorrhagic Fever Virus,CCHFV)中的任意一种或至少两种的组合。Preferably, the virus comprises any one or a combination of at least two of influenza A virus (IAV), vesicular stomatitis virus (VSV), severe fever with thrombocytopenia syndrome virus (SFTSV) or Crimean-Congo hemorrhagic fever virus (CCHFV).

优选地,所述山奈素代谢产物包括山奈酚。Preferably, the kaempferol metabolite comprises kaempferol.

优选地,所述药物还包括干扰素。Preferably, the drug further comprises interferon.

本发明证明了山奈素及其代谢产物和干扰素可以协同增强固有免疫能力。The invention proves that kaempferol and its metabolites and interferon can synergistically enhance the inherent immune ability.

优选地,所述干扰素包括Ⅰ型干扰素和/或Ⅲ型干扰素。Preferably, the interferon includes type I interferon and/or type III interferon.

优选地,所述Ⅰ型干扰素包括IFN-α、IFN-β、IFN-κ、IFN-ε或IFN-ω中的任意一种或至少两种的组合。Preferably, the type I interferon includes any one of IFN-α, IFN-β, IFN-κ, IFN-ε or IFN-ω, or a combination of at least two of them.

优选地,所述Ⅲ型干扰素包括IFN-λ。Preferably, the type III interferon includes IFN-λ.

本发明中提供了Ⅰ型干扰素中的IFN-β参与增强固有免疫,Ⅲ型干扰素的抗病毒功能与I型干扰素重合,因此Ⅲ型干扰素具有与I型干扰素类似的功能是可以合理预期的。The present invention provides that IFN-β in type I interferon participates in enhancing innate immunity, and the antiviral function of type III interferon overlaps with that of type I interferon, so it is reasonable to expect that type III interferon has a function similar to that of type I interferon.

优选地,所述药物的剂型包括片剂、胶囊剂、滴丸剂、溶液剂、气雾剂、喷雾剂、软膏剂或膜剂中的任意一种。Preferably, the dosage form of the drug includes any one of tablets, capsules, pills, solutions, aerosols, sprays, ointments or films.

优选地,所述药物还包括药学上可接受的辅料。Preferably, the drug further comprises a pharmaceutically acceptable excipient.

优选地,所述辅料包括载体、稀释剂、粘合剂、润湿剂、崩解剂、乳化剂、助溶剂、增溶剂、渗透压调节剂、表面活性剂、包衣材料、着色剂、pH调节剂、抗氧剂、抑菌剂或缓冲剂中的任意一种或至少两种的组合。Preferably, the auxiliary materials include any one or a combination of at least two of a carrier, a diluent, a binder, a wetting agent, a disintegrant, an emulsifier, a solubilizer, a solubilizing agent, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, an antibacterial agent or a buffer.

第二方面,本发明提供了山奈素和/或山奈素代谢产物在制备JAK/STAT信号通路激活剂中的应用。In a second aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of a JAK/STAT signaling pathway activator.

优选地,所述JAK/STAT信号通路的激活剂包括I型IFN和/或Ⅲ型IFN。Preferably, the activator of the JAK/STAT signaling pathway includes type I IFN and/or type III IFN.

本发明中证明了山奈素及其代谢产物和干扰素可以协同增强固有免疫能力,可利用山奈素和/或山奈素代谢产物与I型IFN协同,开发活性更强的JAK/STAT信号通路激活剂。The present invention proves that kaempferol and its metabolites can synergistically enhance the innate immune ability with interferon, and kaempferol and/or kaempferol metabolites can be used in synergy with type I IFN to develop a more active JAK/STAT signaling pathway activator.

第三方面,本发明提供了山奈素和/或山奈素代谢产物在制备以非疾病诊断和/或治疗为目的的JAK/STAT信号通路激活剂中的应用。In a third aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of JAK/STAT signaling pathway activators for the purpose of non-disease diagnosis and/or treatment.

本发明中,发现可利用山奈素和/或山奈素代谢产物与I型IFN协同,开发活性更强的JAK/STAT信号通路激活剂,因此可用于制备非疾病治疗等目的的JAK/STAT信号通路激活剂,用于JAK/STAT信号通路的基础行为的研究中等。In the present invention, it is found that kaempferol and/or kaempferol metabolites can be used in conjunction with type I IFN to develop a more active JAK/STAT signaling pathway activator. Therefore, it can be used to prepare a JAK/STAT signaling pathway activator for purposes other than disease treatment, and to study the basic behavior of the JAK/STAT signaling pathway.

第四方面,本发明提供了山奈素和/或山奈素代谢产物在制备增强细胞内I型IFN和/或Ⅲ型IFN的制剂中的应用。In a fourth aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of a preparation for enhancing type I IFN and/or type III IFN in cells.

优选地,所述I型IFN包括IFN-α、IFN-β、IFN-κ、IFN-ε或IFN-ω中的任意一种或至少两种的组合。Preferably, the type I IFN includes any one of IFN-α, IFN-β, IFN-κ, IFN-ε or IFN-ω, or a combination of at least two thereof.

优选地,所述Ⅲ型IFN包括IFN-λ。Preferably, the type III IFN includes IFN-λ.

优选地,所述在细胞内增强I型IFN具体为:延长I型IFN激活JAK/STAT信号通路的时间。Preferably, the enhancement of type I IFN in cells specifically comprises: prolonging the time for type I IFN to activate the JAK/STAT signaling pathway.

第五方面,本发明提供了山奈素和/或山奈素代谢产物在制备以非疾病诊断和/或治疗为目的的增强细胞内I型IFN的制剂中的应用。In a fifth aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of a preparation for enhancing intracellular type I IFN for the purpose of non-disease diagnosis and/or treatment.

本发明中发现山奈素和/或山奈素代谢产物能够延长I型IFN激活JAK/STAT信号通路的时间,可应用于JAK/STAT信号通路的基础行为的研究中等。The present invention finds that kaempferol and/or kaempferol metabolites can prolong the time for type I IFN to activate the JAK/STAT signaling pathway, and can be applied to the study of the basic behavior of the JAK/STAT signaling pathway.

第六方面,本发明提供了山奈素和/或山奈素代谢产物在制备疫苗佐剂、药物缓释制剂、药物控释制剂或靶向给药制剂中的应用。与现有技术相比,本发明具有以下有益效果:In a sixth aspect, the present invention provides the use of kaempferol and/or kaempferol metabolites in the preparation of vaccine adjuvants, drug sustained-release preparations, drug controlled-release preparations or targeted drug delivery preparations. Compared with the prior art, the present invention has the following beneficial effects:

本发明对山奈素及其代谢产物的生理作用进行了研究,研究结果表明,山奈素及其代谢产物山奈酚是一种IFN的应答剂,能够增强和延长I型IFN激活的JAK/STAT通路,与I型IFNs协同作用,增强固有免疫,抑制IAV、VSVG、CCHFV和SFTSV病毒,显现出了广谱的抗病毒潜力。The present invention studies the physiological effects of kaempferol and its metabolites, and the research results show that kaempferol and its metabolite kaempferol are IFN responders, which can enhance and prolong the JAK/STAT pathway activated by type I IFN, synergize with type I IFNs, enhance innate immunity, inhibit IAV, VSVG, CCHFV and SFTSV viruses, and show a broad-spectrum antiviral potential.

附图说明BRIEF DESCRIPTION OF THE DRAWINGS

图1为山奈素和山奈酚的化学结构图;FIG1 is a diagram showing the chemical structures of kaempferol and kaempferol;

图2为鉴定山奈素为I型IFN激活JAK/STAT信号通路的有效增强子结果图,其中,A图为IFN-β处理后ISRE报告基因活性的剂量-反应曲线图,B图为基于ISRE报告的IFN信号调节剂筛选系统的质量控制和信噪比图,C图为黄酮类化合物库的初级筛选JAK/STAT信号通路的激活子或增强子结果图,D图为IFN激活JAK/STAT信号通路的前5个主要命中增强子的验证结果图,E图为山奈素的细胞毒性测试结果图;Figure 2 is a diagram showing the results of identifying kaempferol as an effective enhancer of the JAK/STAT signaling pathway activated by type I IFN, wherein Figure A is a dose-response curve of ISRE reporter gene activity after IFN-β treatment, Figure B is a quality control and signal-to-noise ratio diagram of the IFN signaling regulator screening system based on the ISRE reporter, Figure C is a diagram showing the results of the primary screening of the flavonoid library for activators or enhancers of the JAK/STAT signaling pathway, Figure D is a diagram showing the verification results of the top 5 major hit enhancers of the JAK/STAT signaling pathway activated by IFN, and Figure E is a diagram showing the results of the cytotoxicity test of kaempferol;

图3为山奈素增强I型IFN诱导的细胞抗病毒状态结果图,其中,A图为山奈素IFN-β诱导ISGs表达的结果图,B图为分别使用IAV-Fluc和VSV-GFP评估山奈素抗病毒能力结果图;Figure 3 is a graph showing the results of kaempferol enhancing the antiviral status of cells induced by type I IFN, wherein Figure A is a graph showing the results of kaempferol IFN-β inducing ISGs expression, and Figure B is a graph showing the results of evaluating the antiviral ability of kaempferol using IAV-Fluc and VSV-GFP respectively;

图4为山奈素抗高致病性病毒的抗病毒测定结果图,其中,A图为山奈素和IFN-β处理后SFTSV的S片段相对RNA水平图,B图为山奈素和IFN-β处理后SFTSV的N蛋白表达情况图,C图为山奈素和IFN-β处理后CCHFV的S片段相对RNA水平图,D图为山奈素和IFN-β处理后CCHFV的N蛋白表达情况图;Figure 4 is a graph showing the antiviral assay results of kaempferol against highly pathogenic viruses, wherein Figure A is a graph showing the relative RNA level of the S segment of SFTSV after treatment with kaempferol and IFN-β, Figure B is a graph showing the N protein expression of SFTSV after treatment with kaempferol and IFN-β, Figure C is a graph showing the relative RNA level of the S segment of CCHFV after treatment with kaempferol and IFN-β, and Figure D is a graph showing the N protein expression of CCHFV after treatment with kaempferol and IFN-β;

图5为山奈素对I型IFN激活JAK/STAT信号通路的作用机制分析结果图,其中,A图为山奈素对不同浓度IFN-β诱导的ISRE报告基因活性的影响结果图,B图为山奈素增强IFN-β激活ISRE报告基因活性的时相性结果图,C图为山奈素对IFN-β诱导的STAT1和STAT2磷酸化的结果图,D图为山奈素对IFN-β诱导的SOCS1和SOCS3表达的结果图;Figure 5 is a graph showing the results of the analysis of the mechanism of action of kaempferol on type I IFN activation of the JAK/STAT signaling pathway, wherein Figure A shows the effect of kaempferol on the ISRE reporter gene activity induced by different concentrations of IFN-β, Figure B shows the phase results of kaempferol enhancing the ISRE reporter gene activity activated by IFN-β, Figure C shows the effect of kaempferol on the phosphorylation of STAT1 and STAT2 induced by IFN-β, and Figure D shows the effect of kaempferol on the expression of SOCS1 and SOCS3 induced by IFN-β;

图6为山奈酚增强I型IFN诱导的细胞抗病毒状态结果图,其中,A图为山奈酚对IFN-β刺激的ISRE报告活性结果图,B图为山奈酚的细胞毒性测试结果图,C图为山奈酚对IFN-β诱导的ISG表达结果图,D图为使用IAV-Fluc评估山奈酚抗病毒能力结果图;Figure 6 is a graph showing the results of kaempferol enhancing the antiviral status of cells induced by type I IFN, wherein Figure A is a graph showing the ISRE reporter activity results of kaempferol on IFN-β stimulation, Figure B is a graph showing the cytotoxicity test results of kaempferol, Figure C is a graph showing the results of kaempferol on IFN-β-induced ISG expression, and Figure D is a graph showing the results of using IAV-Fluc to evaluate the antiviral ability of kaempferol;

图中涉及数据的均显示为3次重复的均值±标准差,*,p<0.05,**,p<0.01,***,p<0.001,t检验。The data in the figure are shown as the mean ± standard deviation of 3 replicates, *, p < 0.05, **, p < 0.01, ***, p < 0.001, t-test.

具体实施方式DETAILED DESCRIPTION

为进一步阐述本发明所采取的技术手段及其效果,以下结合实施例和附图对本发明作进一步地说明。可以理解的是,此处所描述的具体实施方式仅仅用于解释本发明,而非对本发明的限定。To further illustrate the technical means and effects of the present invention, the present invention is further described below in conjunction with the embodiments and drawings. It should be understood that the specific implementation methods described herein are only used to explain the present invention, rather than to limit the present invention.

实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可通过正规渠道商购获得的常规产品。If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

本发明中细胞、病毒和抗体等材料的培养方法或来源:The culture methods or sources of the cells, viruses, antibodies and other materials in the present invention are:

人胚胎肾细胞(HEK293T,ATCC,CRL-3216)在补充有10%胎牛血清(FBS)的Dulbecco改良Eagle培养基(DMEM)中培养。人胚胎肾细胞(HEK293,ATCC,CRL-1573)在补充有10%FBS的Eagle's Minimum Essential Medium(EMEM)中培养。所有细胞在37℃和5%CO2下培养。Human embryonic kidney cells (HEK293T, ATCC, CRL-3216) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). Human embryonic kidney cells (HEK293, ATCC, CRL-1573) were cultured in Eagle's Minimum Essential Medium (EMEM) supplemented with 10% FBS. All cells were cultured at 37°C and 5% CO2 .

报告型甲型流感病毒IAV Fluc(PR8-NSCE2-Fluc)和单周期感染性报告型水泡性口腔炎病毒VSV-GFP(VSVΔG/GFP-VG)由本实验室保存。严重发热伴血小板减少综合征病毒(SFTSV)和克里米亚-刚果出血热病毒(CCHFV)由中国科学院病毒学国家重点实验室和生物安全大科学中心提供。The reporter influenza A virus IAV Fluc (PR8-NSCE2-Fluc) and the single-cycle infectious reporter vesicular stomatitis virus VSV-GFP (VSVΔG/GFP-VG) were stored in our laboratory. Severe fever with thrombocytopenia syndrome virus (SFTSV) and Crimean-Congo hemorrhagic fever virus (CCHFV) were provided by the State Key Laboratory of Virology and the Center for Biosafety, Chinese Academy of Sciences.

抗CCHFV NP、抗SFTSV NP、抗β-肌动蛋白、抗-pSTAT1、抗STAT1、抗pSTAT2、抗STAT2、辣根过氧化物酶标记山羊抗小鼠抗体由中国科学院病毒学国家重点实验室和生物安全大科学中心提供。类黄酮文库(HY-L068)及其相关化合物购自MedChemExpress(MCE;Monmouth Junction,NJ,USA)。重组人IFN-β购自Proteintech(Rosemont,IL,USA)。Anti-CCHFV NP, anti-SFTSV NP, anti-β-actin, anti-pSTAT1, anti-STAT1, anti-pSTAT2, anti-STAT2, and horseradish peroxidase-labeled goat anti-mouse antibodies were provided by the State Key Laboratory of Virology and the Center for Biosafety, Chinese Academy of Sciences. Flavonoid library (HY-L068) and its related compounds were purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA). Recombinant human IFN-β was purchased from Proteintech (Rosemont, IL, USA).

实施例1鉴定新型干扰素激活的JAK/STAT信号增强剂Example 1 Identification of novel interferon-activated JAK/STAT signaling enhancers

(1)293T/ISRE-Fluc细胞筛选平台构建:(1) Construction of 293T/ISRE-Fluc cell screening platform:

采用Lipofectamine 2000方案将ISRE启动子荧光素酶报告质粒(pISRE-Luc,2μg/皿/10cm培养皿)转染HEK293T细胞。在5小时后,细胞洗涤和悬浮,然后铺板在2×104细胞/孔的96孔白板中。在18小时后,用不同浓度的IFN-β瞬时转染细胞,在37℃下孵育24小时。然后使用Britelite plus报告基因检测系统(PerkinElmer,Waltham,MA,USA)进行荧光素酶检测。丢弃96孔板上生长的细胞培养液,依次加入50μL PBS和50μL底物。振荡10min后,立即使用BioTek SYNERGY neo2酶标仪(BioTek,Winooski,VT,USA)检测发光。HEK293T cells were transfected with ISRE promoter luciferase reporter plasmid (pISRE-Luc, 2 μg/dish/10 cm culture dish) using Lipofectamine 2000 protocol. After 5 hours, cells were washed and suspended, and then plated in 96-well white plates at 2×10 4 cells/well. After 18 hours, cells were transiently transfected with different concentrations of IFN-β and incubated at 37°C for 24 hours. Luciferase assays were then performed using the Britelite plus reporter gene assay system (PerkinElmer, Waltham, MA, USA). The cell culture medium grown on the 96-well plate was discarded, and 50 μL PBS and 50 μL substrate were added in sequence. After shaking for 10 min, luminescence was immediately detected using a BioTek SYNERGY neo2 microplate reader (BioTek, Winooski, VT, USA).

(2)鉴定JAK/STAT激活剂:(2) Identification of JAK/STAT activators:

从黄酮类化合物库中提取的5μM化合物分别处理瞬时转染的HEK293T细胞,并在37℃孵育24小时,然后进行荧光素酶检测。Transiently transfected HEK293T cells were treated with 5 μM of compounds extracted from the flavonoid library and incubated at 37 °C for 24 h, followed by luciferase assay.

(3)鉴定I型IFN激活JAK/STAT通路增强子:(3) Identification of enhancers of JAK/STAT pathway activated by type I IFN:

用6ng/mL的IFN-β瞬时转染HEK293T细胞,并结合5μM的黄酮类化合物库,孵育24h后进行荧光素酶检测。HEK293T cells were transiently transfected with 6 ng/mL IFN-β and combined with 5 μM flavonoid library, and luciferase assay was performed after incubation for 24 h.

如图2中A图所示,IFN-β处理以剂量依赖性方式刺激ISRE介导的Fluc表达。293T/ISRE-Fluc细胞成功被改造为JAK/STAT信号通路激动剂的筛选平台。As shown in Figure 2A, IFN-β treatment stimulated ISRE-mediated Fluc expression in a dose-dependent manner. 293T/ISRE-Fluc cells were successfully transformed into a screening platform for JAK/STAT signaling pathway agonists.

如图2中B图所示,在6pg/mL的IFN-β存在的情况下293T/ISRE-Fluc细胞被改造成鉴定I型IFN激活JAK/STAT通路增强子的筛选平台,6pg/mL的IFN-β驱动可检测但远未达到饱和水平的Fluc表达,产生的理想信噪比(S/N)和Z'值分别为38和0.56。As shown in Figure 2B, 293T/ISRE-Fluc cells were transformed into a screening platform to identify enhancers of type I IFN-activated JAK/STAT pathway in the presence of 6 pg/mL IFN-β, which drove detectable but far from saturated levels of Fluc expression, producing ideal signal-to-noise ratios (S/N) and Z' values of 38 and 0.56, respectively.

如图2中C图所示,在IFN-β存在的情况下,黄酮类化合物处理的293T/ISRE-Fluc细胞的中位信号显著升高,而在IFN-β不存在的情况下,中位信号与对照组相当。这些结果表明,类黄酮增强I型IFN激活的JAK/STAT通路。As shown in Figure 2, Panel C, in the presence of IFN-β, the median signal of 293T/ISRE-Fluc cells treated with flavonoids was significantly increased, while in the absence of IFN-β, the median signal was comparable to that of the control group. These results suggest that flavonoids enhance the JAK/STAT pathway activated by type I IFN.

如图2中D图所示,选择能够增强IFN激活的ISRE报告基因表达的前5个化合物进行剂量-反应分析。结果显示5个命中物均具有剂量依赖性,其中山奈素显示出比其他更强的潜力。As shown in Figure 2D, the top five compounds that could enhance the expression of the ISRE reporter gene activated by IFN were selected for dose-response analysis. The results showed that all five hits were dose-dependent, with kaempferol showing a stronger potential than the others.

如图2中E图所示,山奈素即使在50μM浓度时也没有表现出细胞毒性。As shown in Figure 2, Panel E, kaempferol showed no cytotoxicity even at a concentration of 50 μM.

实施例2山奈素对IFN应答的典型抗病毒基因ISGs的影响Example 2 Effect of Kaempferol on Typical Antiviral Genes ISGs of IFN Response

在24孔板中,分别用6pg/mL IFN-β单独或与试验化合物联合处理HEK293T细胞。24h后,提取细胞总RNA,使用PrimeScriptTMRT Master Mix(Takara,日本)进行逆转录,进行RT-qPCR分析。RT-qPCR采用TB Green Premix Ex Taq II试剂盒(Takara,日本),CFXConnect实时PCR检测系统(Bio-Rad,德国)进行。使用以下程序分析这些样品:95℃,30s;95℃,5s,60℃,30s,40个循环。采用2-ΔΔCT法计算ISGs的相对表达量。以GAPDH为内参照。用于qPCR分析的引物列于表1。HEK293T cells were treated with 6 pg/mL IFN-β alone or in combination with test compounds in 24-well plates. After 24 h, total cellular RNA was extracted and reverse transcribed using PrimeScript TM RT Master Mix (Takara, Japan) for RT-qPCR analysis. RT-qPCR was performed using the TB Green Premix Ex Taq II Kit (Takara, Japan) and the CFXConnect Real-Time PCR Detection System (Bio-Rad, Germany). The samples were analyzed using the following program: 95°C, 30 s; 95°C, 5 s, 60°C, 30 s, 40 cycles. The relative expression of ISGs was calculated using the 2-ΔΔCT method. GAPDH was used as an internal reference. The primers used for qPCR analysis are listed in Table 1.

表1Table 1

如图3中A图所示,以6pg/mL的IFN-β处理HEK293T细胞,与DMSO对照相比,10μM的山奈素提高了多种ISGs的表达,包括ISG15、MxA、ISG20、IFIT1和IFITM3等。As shown in Figure 3A, HEK293T cells were treated with 6 pg/mL IFN-β, and 10 μM kaempferol increased the expression of multiple ISGs, including ISG15, MxA, ISG20, IFIT1, and IFITM3, compared with the DMSO control.

实施例3山奈素抗IAV和VSV病毒活性测定Example 3 Determination of the anti-IAV and VSV viral activity of kaempferol

用不同浓度的IFN-β处理HEK293T细胞,分别加入或不加入2或10μM山奈素联合处理HEK293T细胞。24小时后,去除含有IFN-β/山奈素的培养基,分别用报告病毒IAV-Fluc或VSV-GFP感染细胞。8小时后,应用荧光素酶法和荧光成像法测定IAV-Fluc和VSV-GFP的感染性。HEK293T cells were treated with different concentrations of IFN-β, with or without 2 or 10 μM kaempferol. After 24 hours, the medium containing IFN-β/kaempferol was removed and the cells were infected with the reporter virus IAV-Fluc or VSV-GFP, respectively. After 8 hours, the infectivity of IAV-Fluc and VSV-GFP was determined by luciferase assay and fluorescence imaging.

结果如图3中B图所示,预先用IFN-β处理细胞以剂量依赖性方式降低了IAV-Fluc和VSV-GFP的感染性,山奈素进一步增强了抗病毒活性。表明山奈素可以通过增强JAK/STAT信号通路特异性地增强I型IFN的抗病毒活性。The results are shown in Figure 3B. Pre-treatment of cells with IFN-β reduced the infectivity of IAV-Fluc and VSV-GFP in a dose-dependent manner, and kaempferol further enhanced the antiviral activity. This indicates that kaempferol can specifically enhance the antiviral activity of type I IFN by enhancing the JAK/STAT signaling pathway.

实施例4山奈素对高致病性病毒SFTSV和CCHFV的抗病毒活性Example 4 Antiviral activity of kaempferol against highly pathogenic viruses SFTSV and CCHFV

用不同浓度的IFN-β单独或与山奈素联合预处理HEK293细胞。用MOI为0.05TCID50/细胞的SFTSV或者CCHFV感染细胞2h后,再加入山奈素孵育24h。细胞在含有蛋白酶抑制剂(Solarbio,北京,中国)的RIPA裂解缓冲液中溶解,蛋白质样品在10%的SDS-PAGE凝胶上分离。将蛋白转移到聚偏二氟乙烯(PVDF)膜上进行Western blot分析。用5%BSA在TBST中封闭PVDF膜后,将PVDF膜依次与一抗和辣根过氧化物酶偶联的相应二抗孵育。采用增强化学发光(ECL)试剂盒(Thermo Fisher,Carlsbad,CA,USA)检测蛋白条带。RT-qPCR步骤参照实施例2进行。HEK293 cells were pretreated with different concentrations of IFN-β alone or in combination with kaempferol. Cells were infected with SFTSV or CCHFV at an MOI of 0.05 TCID50/cell for 2 h, and then kaempferol was added for incubation for 24 h. Cells were dissolved in RIPA lysis buffer containing protease inhibitors (Solarbio, Beijing, China), and protein samples were separated on 10% SDS-PAGE gels. The proteins were transferred to polyvinylidene fluoride (PVDF) membranes for Western blot analysis. After blocking the PVDF membrane with 5% BSA in TBST, the PVDF membrane was incubated with primary antibodies and corresponding secondary antibodies coupled to horseradish peroxidase in turn. Protein bands were detected using an enhanced chemiluminescence (ECL) kit (Thermo Fisher, Carlsbad, CA, USA). The RT-qPCR steps were performed with reference to Example 2.

如图4中A图和B图所示,IFN-β处理以剂量依赖性方式抑制SFTSV S段RNA的积累,而山奈素进一步增强了不同浓度IFN-β的抗病毒活性。此外,山奈素单独治疗也显示出一定的抑制作用,可能是通过增强内源性IFNs驱动的JAK/STAT信号通路在病毒感染中的作用。As shown in Figure 4A and B, IFN-β treatment inhibited the accumulation of SFTSV S segment RNA in a dose-dependent manner, and kaempferol further enhanced the antiviral activity of different concentrations of IFN-β. In addition, kaempferol treatment alone also showed a certain inhibitory effect, which may be through enhancing the role of the JAK/STAT signaling pathway driven by endogenous IFNs in viral infection.

如图4中C图和D图所示,山奈素单独使用可以在一定程度上抑制CCHFV S片段RNA的复制以及病毒NP蛋白的表达,山奈素和IFN-β联合使用显示出了协同作用。As shown in Figure 4 C and D, kaempferol alone can inhibit the replication of CCHFV S segment RNA and the expression of viral NP protein to a certain extent, and the combined use of kaempferol and IFN-β shows a synergistic effect.

实施例5山奈素增强IFN信号传导的机制研究Example 5 Study on the mechanism of kaempferol enhancing IFN signal transduction

在24孔板中,分别用不同浓度的IFN-β与10μM山奈素联合处理转染ISRE的HEK293T细胞。孵育24h后进行荧光素酶检测。采用同样处理方法,在不同时间段检测ISRE报告基因的活性。In a 24-well plate, different concentrations of IFN-β and 10 μM kaempferol were used to treat HEK293T cells transfected with ISRE. Luciferase assay was performed after 24 hours of incubation. The activity of the ISRE reporter gene was detected at different time periods using the same treatment method.

如图5中A图所示,山奈素的活性与IFN-β的浓度无关,饱和IFN-β诱导的平台期信号也显著升高,表明山奈素不可能通过增加IFN-β与IFNR的亲和力发挥作用。As shown in Figure 5A, the activity of kaempferol was independent of the concentration of IFN-β, and the plateau signal induced by saturated IFN-β was also significantly increased, indicating that kaempferol could not exert its effect by increasing the affinity of IFN-β to IFNR.

如图5中B图所示,ISRE报告基因的活性在IFN-β处理后12小时达到峰值,然后迅速下降,而山奈酚直到IFN-β处理后12小时才影响ISRE报告基因的表达。此外,山奈素处理在IFN-β处理后的12-24小时内,信号保持在相对较高的水平而没有减弱,表明山奈素延长了IFN-β刺激JAK/STAT信号的持续时间。As shown in Figure 5B, the activity of the ISRE reporter gene peaked 12 hours after IFN-β treatment and then rapidly decreased, while kaempferol did not affect the expression of the ISRE reporter gene until 12 hours after IFN-β treatment. In addition, the signal remained at a relatively high level without attenuation within 12-24 hours after IFN-β treatment, indicating that kaempferol prolonged the duration of IFN-β stimulation of JAK/STAT signaling.

实施例6山奈素抑制SOCS3介导的负反馈延长I型IFN激活的JAK/STAT信号传导Example 6 Kaempferol inhibits SOCS3-mediated negative feedback and prolongs JAK/STAT signaling activated by type I IFN

用20μM山奈素或DMSO对照处理HEK293细胞,同时加入或不加入IFN-β(5ng/mL)。在处理后6小时,收集细胞,裂解并进行蛋白质印迹分析,蛋白质印迹分析步骤参照实施例4进行。HEK293 cells were treated with 20 μM kaempferol or DMSO control, with or without the addition of IFN-β (5 ng/mL). Six hours after treatment, cells were collected, lysed and subjected to Western blot analysis, and the Western blot analysis procedure was performed as described in Example 4.

细胞因子信号转导抑制因子1和3(SOCS1和SOCS3)是两个典型的IFN信号的反馈抑制因子,检测了SOCS1和SOCS3在IFN-β联合或不联合山奈素处理下的表达水平,RT-qPCR步骤参照实施例2进行。Suppressor of cytokine signaling 1 and 3 (SOCS1 and SOCS3) are two typical feedback inhibitors of IFN signaling. The expression levels of SOCS1 and SOCS3 under IFN-β combined with or without kaempferol treatment were detected. The RT-qPCR step was performed with reference to Example 2.

如图5中C图所示,山奈酚的加入对IFN-β激活的ISRE报告基因表达没有影响,STAT1和STAT2的磷酸化程度没有显著增加。As shown in Figure 5C, the addition of kaempferol had no effect on the expression of the ISRE reporter gene activated by IFN-β, and the phosphorylation levels of STAT1 and STAT2 were not significantly increased.

如图5中D图所示,无论是在基线水平还是在IFN-β诱导水平,山奈素均不影响SOCS1的表达水平,显著下调SOCS3的表达水平。山奈素可能通过抑制SOCS3介导的负反馈延长IFN激活的JAK/STAT信号通路。As shown in Figure 5D, kaempferol did not affect the expression level of SOCS1 at both the baseline level and the IFN-β-induced level, but significantly downregulated the expression level of SOCS3. Kaempferol may prolong the IFN-activated JAK/STAT signaling pathway by inhibiting the negative feedback mediated by SOCS3.

实施例7山奈素代谢产物山奈酚的抗病毒活性测定Example 7 Determination of the antiviral activity of kaempferol, a metabolite of kaempferol

在体内给药时,山奈素可能部分被肝微粒体氧化为山奈酚,因此检测了山奈酚对I型IFN驱动的ISG表达和细胞抗病毒状态的影响,实验步骤参照实施例2、3和4进行。When administered in vivo, kaempferol may be partially oxidized to kaempferol by liver microsomes, so the effect of kaempferol on type I IFN-driven ISG expression and cellular antiviral status was detected. The experimental procedures were carried out with reference to Examples 2, 3 and 4.

如图6中A图所示,山奈酚以剂量依赖性方式显著增加了IFN-β刺激的ISRE启动子活性。As shown in Figure 6A, kaempferol significantly increased IFN-β-stimulated ISRE promoter activity in a dose-dependent manner.

如图6中B图所示,山奈酚即使在50μM浓度时也没有表现出细胞毒性。As shown in Figure 6B, kaempferol showed no cytotoxicity even at a concentration of 50 μM.

如图6中C图所示,RT-qPCR分析表明山奈酚处理增加了IFN-β诱导的经典抗病毒ISGs表达量,ISG15和MxA的表达增加了约4倍。As shown in Figure 6C, RT-qPCR analysis showed that kaempferol treatment increased the expression of IFN-β-induced classical antiviral ISGs, and the expression of ISG15 and MxA increased by about 4 times.

如图6中D图所示,山奈酚显著增强了IFN-β对IAV-Fluc的抗病毒活性。As shown in Figure 6D, kaempferol significantly enhanced the antiviral activity of IFN-β against IAV-Fluc.

综上所述,本发明对山奈素及其代谢产物的生理作用进行了研究,研究结果表明,山奈素及其代谢产物山奈酚是一种IFN的应答剂,能够增强和延长I型IFN激活的JAK/STAT通路,与I型IFNs协同作用,增强固有免疫,抑制IAV、VSVG、CCHFV和SFTSV病毒,显现出了广谱的抗病毒潜力。In summary, the present invention studies the physiological effects of kaempferol and its metabolites, and the research results show that kaempferol and its metabolite kaempferol are IFN responders, which can enhance and prolong the JAK/STAT pathway activated by type I IFN, synergize with type I IFNs, enhance innate immunity, inhibit IAV, VSVG, CCHFV and SFTSV viruses, and show a broad-spectrum antiviral potential.

申请人声明,以上所述仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,所属技术领域的技术人员应该明了,任何属于本技术领域的技术人员在本发明揭露的技术范围内,可轻易想到的变化或替换,均落在本发明的保护范围和公开范围之内。The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention shall fall within the protection scope and disclosure scope of the present invention.

Claims (10)

1.山奈素和/或山奈素代谢产物在制备抗病毒感染的药物中的应用。1. Use of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection. 2.根据权利要求1所述的应用,其特征在于,所述病毒包括DNA病毒和/或RNA病毒;2. The use according to claim 1, characterized in that the virus comprises a DNA virus and/or an RNA virus; 优选地,所述DNA病毒包括乙肝病毒、疱疹病毒、人乳头瘤病毒或腺病毒中的任意一种或至少两种的组合;Preferably, the DNA virus comprises any one or a combination of at least two of hepatitis B virus, herpes virus, human papillomavirus or adenovirus; 优选地,所述RNA病毒包括艾滋病病毒、SARS病毒、MERS病毒、埃博拉病毒、流感病毒、副流感病毒、狂犬病毒、腮腺炎病毒、麻疹病毒、呼吸道合胞病毒、艾柯病毒、黄病毒、甲病毒、布尼亚病毒、甲型肝炎病毒、柯萨奇病毒、鼻病毒、肠道病毒、脊髓灰质炎病毒、登革热病毒、轮状病毒、马尔堡病毒或新型冠状病毒中的任意一种或至少两种的组合;Preferably, the RNA virus comprises any one or a combination of at least two of HIV, SARS, MERS, Ebola, influenza, parainfluenza, rabies, mumps, measles, respiratory syncytial virus, echovirus, flavivirus, alphavirus, bunyavirus, hepatitis A, coxsackievirus, rhinovirus, enterovirus, polio, dengue, rotavirus, Marburg or novel coronavirus; 优选地,所述病毒包括甲型流感病毒、水泡性口腔炎病毒、严重发热伴血小板减少综合征病毒或克里米亚-刚果出血热病毒中的任意一种或至少两种的组合;Preferably, the virus comprises any one or a combination of at least two of influenza A virus, vesicular stomatitis virus, severe fever with thrombocytopenia syndrome virus or Crimean-Congo hemorrhagic fever virus; 优选地,所述山奈素代谢产物包括山奈酚。Preferably, the kaempferol metabolite comprises kaempferol. 3.根据权利要求1或2所述的应用,其特征在于,所述药物还包括干扰素;3. The use according to claim 1 or 2, characterized in that the drug further comprises interferon; 优选地,所述干扰素包括Ⅰ型干扰素和/或Ⅲ型干扰素;Preferably, the interferon includes type I interferon and/or type III interferon; 优选地,所述Ⅰ型干扰素包括IFN-α、IFN-β、IFN-κ、IFN-ε或IFN-ω中的任意一种或至少两种的组合;Preferably, the type I interferon includes any one of IFN-α, IFN-β, IFN-κ, IFN-ε or IFN-ω, or a combination of at least two thereof; 优选地,所述Ⅲ型干扰素包括IFN-λ。Preferably, the type III interferon includes IFN-λ. 4.根据权利要求1~3任一项所述的应用,其特征在于,所述药物的剂型包括片剂、胶囊剂、滴丸剂、溶液剂、气雾剂、喷雾剂、软膏剂或膜剂中的任意一种。4. The use according to any one of claims 1 to 3, characterized in that the dosage form of the drug includes any one of tablets, capsules, pills, solutions, aerosols, sprays, ointments or films. 5.根据权利要求1~4任一项所述的应用,其特征在于,所述药物还包括药学上可接受的辅料;5. The use according to any one of claims 1 to 4, characterized in that the drug further comprises a pharmaceutically acceptable excipient; 优选地,所述辅料包括载体、稀释剂、粘合剂、润湿剂、崩解剂、乳化剂、助溶剂、增溶剂、渗透压调节剂、表面活性剂、包衣材料、着色剂、pH调节剂、抗氧剂、抑菌剂或缓冲剂中的任意一种或至少两种的组合。Preferably, the auxiliary materials include any one or a combination of at least two of a carrier, a diluent, a binder, a wetting agent, a disintegrant, an emulsifier, a solubilizer, a solubilizing agent, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, an antibacterial agent or a buffer. 6.山奈素和/或山奈素代谢产物在制备JAK/STAT信号通路激活剂中的应用。6. Use of kaempferol and/or kaempferol metabolites in the preparation of JAK/STAT signaling pathway activators. 7.根据权利要求6所述的应用,其特征在于,所述JAK/STAT信号通路的激活剂包括I型IFN和/或Ⅲ型IFN。7. The use according to claim 6, characterized in that the activator of the JAK/STAT signaling pathway comprises type I IFN and/or type III IFN. 8.山奈素和/或山奈素代谢产物在制备增强细胞内I型IFN和/或Ⅲ型IFN的制剂中的应用。8. Use of kaempferol and/or kaempferol metabolites in the preparation of preparations for enhancing intracellular type I IFN and/or type III IFN. 9.根据权利要求8所述的应用,其特征在于,所述I型IFN包括IFN-α、IFN-β、IFN-κ、IFN-ε或IFN-ω中的任意一种或至少两种的组合;9. The use according to claim 8, characterized in that the type I IFN comprises any one of IFN-α, IFN-β, IFN-κ, IFN-ε or IFN-ω, or a combination of at least two thereof; 优选地,所述Ⅲ型IFN包括IFN-λ;Preferably, the type III IFN includes IFN-λ; 优选地,所述在细胞内增强I型IFN具体为:延长I型IFN激活JAK/STAT信号通路的时间。Preferably, the enhancement of type I IFN in cells specifically comprises: prolonging the time for type I IFN to activate the JAK/STAT signaling pathway. 10.山奈素和/或山奈素代谢产物在制备疫苗佐剂、药物缓释制剂、药物控释制剂或靶向给药制剂中的应用。10. Use of kaempferol and/or kaempferol metabolites in the preparation of vaccine adjuvants, drug sustained-release preparations, drug controlled-release preparations or targeted drug delivery preparations.
CN202410829865.2A 2024-06-25 2024-06-25 Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection Pending CN118845751A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202410829865.2A CN118845751A (en) 2024-06-25 2024-06-25 Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202410829865.2A CN118845751A (en) 2024-06-25 2024-06-25 Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection

Publications (1)

Publication Number Publication Date
CN118845751A true CN118845751A (en) 2024-10-29

Family

ID=93162035

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202410829865.2A Pending CN118845751A (en) 2024-06-25 2024-06-25 Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection

Country Status (1)

Country Link
CN (1) CN118845751A (en)

Similar Documents

Publication Publication Date Title
Liu et al. The C-terminal tail of TRIM56 dictates antiviral restriction of influenza A and B viruses by impeding viral RNA synthesis
Chan et al. Broad-spectrum antivirals for the emerging Middle East respiratory syndrome coronavirus
Liu et al. Overlapping and distinct molecular determinants dictating the antiviral activities of TRIM56 against flaviviruses and coronavirus
Pyrc et al. Inhibition of human coronavirus NL63 infection at early stages of the replication cycle
Cheung et al. Cytokine responses in severe acute respiratory syndrome coronavirus-infected macrophages in vitro: possible relevance to pathogenesis
Villenave et al. Induction and antagonism of antiviral responses in respiratory syncytial virus-infected pediatric airway epithelium
US8232240B2 (en) Inhibitors of enveloped virus infectivity
Scagnolari et al. Increased sensitivity of SARS-coronavirus to a combination of human type I and type II interferons
Haasbach et al. Low-dose interferon Type I treatment is effective against H5N1 and swine-origin H1N1 influenza A viruses in vitro and in vivo
Nakamichi et al. Rabies virus-induced activation of mitogen-activated protein kinase and NF-κB signaling pathways regulates expression of CXC and CC chemokine ligands in microglia
Lieskovska et al. Tick salivary cystatin sialostatin L2 suppresses IFN responses in mouse dendritic cells
JP2023526327A (en) Compositions and methods for enhancing drug efficacy
Bradel-Tretheway et al. Nipah and Hendra virus glycoproteins induce comparable homologous but distinct heterologous fusion phenotypes
Harris et al. The multifaceted roles of NLRP3-modulating proteins in virus infection
KR20230004636A (en) Bidofludimus for use in the treatment or prevention of viral diseases
Cinatl Jr et al. Role of interferons in the treatment of severe acute respiratory syndrome
CN108712905A (en) Anti-hepatic tumor virus agent
Bhattacharyya Inflammation during virus infection: swings and roundabouts
Li et al. IFN-λs inhibit Hantaan virus infection through the JAK-STAT pathway and expression of Mx2 protein
Ramírez-Carvajal et al. Constitutively active IRF7/IRF3 fusion protein completely protects swine against foot-and-mouth disease
CN118845751A (en) Application of kaempferol and/or kaempferol metabolites in the preparation of drugs for antiviral infection
Du et al. Kaempferide enhances type I interferon signaling as a novel broad-spectrum antiviral agent
US20220323471A1 (en) Glycosylated diphyllin as a broad-spectrum antiviral agent against zika virus and covid-19
Uddin et al. Potential drugs for the treatment of COVID-19: Synthesis, brief history and application
EP4149468A1 (en) Compound for preventing or treating a viral infection

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination