WO2023197484A1 - 马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用 - Google Patents

马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用 Download PDF

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WO2023197484A1
WO2023197484A1 PCT/CN2022/111333 CN2022111333W WO2023197484A1 WO 2023197484 A1 WO2023197484 A1 WO 2023197484A1 CN 2022111333 W CN2022111333 W CN 2022111333W WO 2023197484 A1 WO2023197484 A1 WO 2023197484A1
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histamine
pneumonia
iva
aristolochic acid
application according
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PCT/CN2022/111333
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French (fr)
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梁爱华
王连嵋
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中国中医科学院中药研究所
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Priority to US18/226,226 priority Critical patent/US20230372288A1/en
Publication of WO2023197484A1 publication Critical patent/WO2023197484A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A61K31/36—Compounds containing methylenedioxyphenyl groups, e.g. sesamin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • Y—GENERAL 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • the invention belongs to the technical field of new medical uses, and specifically relates to the application of aristolochic acid IVa in the preparation of antihistamine drugs or drugs for treating pneumonia.
  • Histamine is an allergic and inflammatory mediator. Many tissues, especially mast cells in the skin, lungs, and intestinal mucosa, contain large amounts of histamine. Histamine is released when tissue is damaged or when inflammation and allergic reactions occur. Histamine has a strong vasodilatory effect and can increase the permeability of the walls of capillaries and venules, causing plasma to leak into tissues, causing local tissue edema. Clinically, many diseases are related to histamine release, such as skin allergies, rhinitis, asthma, anaphylactic shock, etc.
  • Pneumonia is caused by a variety of pathogens (such as viruses, bacteria, fungi, parasites), radiation, chemicals, allergies and other factors.
  • Typical pneumonia is lobar pneumonia or bronchopneumonia caused by common bacteria such as Streptococcus pneumoniae.
  • SARS is a pneumonia caused by mycoplasma, chlamydia, legionella, rickettsia, viruses and other unknown microorganisms. Among them, SARS virus and Middle East respiratory syndrome virus (MERS) can cause severe pneumonia. Edema and multiple organ failure.
  • Acute pulmonary edema is an important cause of patient death caused by acute lung infection and chemical lung injury (chlorine or phosgene poisoning).
  • Aristolochic acids are nitrophenanthrenecarboxylic acid compounds.
  • aristolochic acid was considered to be nephrotoxic and carcinogenic.
  • the toxic aristolochic acids are mainly AA-I and AA-II.
  • AA-IVa aristolochic acid IVa
  • the object of the present invention is to provide a new medical use of aristolochic acid IVa, specifically to provide a method for preparing antihistamine drugs or drugs for treating pneumonia using aristolochic acid IVa and its pharmaceutically acceptable salts as active components. applications in.
  • aristolochic acid IVa in the preparation of antihistamine drugs or drugs for the treatment of pneumonia.
  • the molecular formula of aristolochic acid IVa is: C17H11NO8, and the chemical structural formula is:
  • the use of the antihistamine drug includes use for skin allergies, eczema, dermatitis, rhinitis or asthma.
  • the pneumonia includes pneumonia caused by pathogens, radiation, chemicals, and allergic factors and atypical pneumonia.
  • the pathogen includes at least one of viruses, bacteria, fungi and parasites.
  • the atypical pneumonia is caused by at least one of SARS, MERS virus, novel coronavirus, adenovirus, influenza virus and Legionella.
  • the medicine contains aristolochic acid IVa as the active ingredient and a pharmaceutically acceptable carrier.
  • the carrier includes at least one of a diluent, a binder, an absorbent, a disintegrant, a dispersant, a wetting agent, a co-solvent, a buffer and a surfactant.
  • the drug is administered by at least one of oral, sublingual, oral mucosa, intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, nasal and rectal routes.
  • the drug is in solid, liquid or gas form.
  • the solid form is a powder, tablet, granule, pill, hard or soft capsule, cream, ointment, plaster, gel, paste, powder or patch.
  • the liquid form is a solution, suspension, injection, syrup, liniment, emulsion, tincture or elixir.
  • the gas form is: aerosol or spray.
  • Another object of the present invention is to provide an antihistamine drug containing the active ingredient aristolochic acid IVa or a medically acceptable salt thereof.
  • Another object of the present invention is to provide a medicine for treating pneumonia, which contains the active ingredient aristolochic acid IVa or a medically acceptable salt thereof.
  • the aristolochic acid IVa of the present invention can significantly inhibit the increase in vascular permeability induced by histamine and exhibit significant antihistamine effects. At the same time, the aristolochic acid IVa can significantly reduce the amount of inflammatory exudation in lung tissue.
  • the aristolochic acid IVa of the present invention achieves antihistamine effects by inhibiting vascular permeability, and at the same time, the aristolochic acid IVa treats pneumonia by reducing the amount of tissue inflammation exuded.
  • aristolochic acid IVa in the present invention has no obvious toxicity and is safe, which provides a new direction for the application of traditional Chinese medicines containing aristolochic acid.
  • Figure 1 shows that AA-IVa reduces the increase in vascular permeability induced by histamine (shown as auricularia staining).
  • Figure 2 shows that AA-IVa reduces inflammatory exudation in lung tissue (HE pathology picture).
  • Figure 3 shows the effects of AA-I and AA-II on histamine-induced increase in vascular permeability.
  • Figure 4 shows the effect of AA-I and AA-II on inflammatory exudation in lung tissue (HE pathology picture).
  • Figure 5 is a comparison of the renal toxicity of AA-I, AA-II and AA-IVa (kidney pathology pictures).
  • A is the control group
  • B is the histamine group
  • C is the histamine+AA-IVa/low-dose group
  • D is the histamine+AA-IVa/high-dose group
  • E is the histamine+AA-I group
  • F It is the histamine + AA-II group.
  • Example 1 Effect of AA-IVa on histamine-induced increase in vascular permeability in mice
  • Animal weight Select 23-25g animals, and start administering the test substance after 1 day of adaptive breeding.
  • Source Beijing Weitonglihua Experimental Animal Technology Co., Ltd.
  • Animal requirements no special pathogenic bacteria.
  • the animal breeding location during the experiment Use the barrier environment animal laboratory of the Institute of Traditional Chinese Medicine, Chinese Academy of Chinese Medical Sciences.
  • Raising conditions barrier system, temperature 20-26°C, relative humidity 40-70%, fresh air. Adopt artificial lighting with a 12-hour light-dark cycle. Animals were housed in polycarbonate mouse cages with 5 mice per cage.
  • Feed Use standard mouse pellet feed, provided by Beijing Keao Xieli Feed Co., Ltd.
  • Drinking water Drink purified water and replace autoclaved drinking bottles twice a week.
  • Microplate reader Thermo, USA, model, VARIOSKAN LUX.
  • ICR mice were divided into 6 groups, including the negative control group (control): they were given normal saline by gavage for 3 days, and normal saline was given tail vein injection on the 4th day; histamine model group (histamine): they were given normal saline by gavage for 3 days.
  • AA-IVa low-dose treatment group (histamine + AA-IVa/L): give AA-IVa 5mg/kg orally for 3 days, and give histamine on the 4th day 5 mg/kg tail vein injection
  • AA-IVa high-dose treatment group (histamine + AA-IVa/H): AA-IVa 10 mg/kg was given by gavage for 3 days, and histamine 5 mg/kg was given by tail vein injection on the 4th day.
  • AA-IVa Use 0.5wt% CMC-Na configuration, the concentrations are 0.5mg/ml and 0.25mg/ml respectively.
  • Histamine Prepare a 0.5 mg/ml histamine solution in physiological saline.
  • Histamine can cause the permeability of the walls of capillaries and venules to increase, causing plasma to leak into tissues, leading to local tissue edema.
  • Evans blue (EB) is an indicator commonly used to detect vascular permeability. When mice were intravenously injected with a solution of histamine and EB, due to the increase in vascular permeability caused by histamine, it was observed that the vascular permeability of the mouse auricle increased (the auricle was stained blue).
  • Auricle vascular permeability test (EB leakage test): After the above-mentioned auricle blue dye observation and photography, cut off both ears and mince them into pieces. After soaking in 2 ml of formamide for 48 hours at room temperature, filter with a 200 mesh stainless steel mesh. Take the filtrate and measure the absorbance (A) at 610nm. According to the Evans blue standard curve, calculate the amount of Evans blue dye exudation ( ⁇ g).
  • EB exudation reduction rate (%) (average EB exudation in the histamine group - average EB exudation in the administration group)/average EB exudation in the histamine group ⁇ 100%.
  • ICR mice were divided into 4 groups, including (1) negative control group (control): given normal saline intragastrically for 3 days, and given normal saline tail vein injection on the 4th day; (2) histamine model group (histamine): given Normal saline was administered by gavage for 3 days, and histamine 5 mg/kg was injected into the tail vein on the fourth day; (3) AA-I treatment group (histamine + AA-I): AA-I 10 mg/kg was administered by gavage for 3 days, and the Histamine 5 mg/kg was injected into the tail vein on the 4th day; (4) AA-II treatment group (histamine + AA-II): AA-II 10 mg/kg was given by gavage for 3 days, and histamine 5 mg/kg was given on the 4th day.
  • Control negative control
  • histamine model group histamine
  • AA-I treatment group AA-I 10 mg/kg was administered by gavage for 3 days, and the Histamine 5
  • Preparation of AA-I and AA-II Use 0.5wt% CMC-Na to prepare AA-I and AA-II to 0.5mg/ml respectively.
  • Preparation of histamine Use physiological saline to prepare a 0.5mg/ml histamine solution.
  • Example 2 AA-IVa relieves histamine-induced pneumonia
  • Figure 2A shows the normal control group, showing that the alveolar and tracheal structures of the lung tissue are normal.
  • Figure 2B shows the model control group, which shows thickening of the alveolar septa and increased infiltration of inflammatory cells.
  • Figure 2C shows the histamine + AA-IVa/L group, which shows thickening of the alveolar septa and increased infiltration of inflammatory cells, but this is significantly reduced compared with the model group.
  • Figure 2D shows the histamine + AA-IVa/H group, which shows thickening of the alveolar septa and increased infiltration of inflammatory cells, but this is significantly reduced compared with the model group.
  • Figure 4 shows the pathological results of lung tissue:
  • Figure 4A shows the normal control group, showing that the alveolar and tracheal structures of the lung tissue are normal.
  • Figure 4B shows the histamine model group, which shows thickening of the alveolar septa and increased infiltration of inflammatory cells, indicating obvious pneumonia.
  • Figure 4E shows the AA-I treatment group, which shows thickening of the alveolar septa and a significant increase in inflammatory cell infiltration. There is no significant difference compared with the histamine model group, suggesting that AA-I treatment has no significant improvement in histamine-induced pneumonia.
  • Figure 4F shows the AA-II group, which shows thickening of the alveolar septa and increased infiltration of inflammatory cells. There is no significant difference compared with the histamine model group, suggesting that AA-II treatment has no significant improvement in histamine-induced pneumonia.
  • mice were randomly divided into 4 groups: (1) control group (administered with normal saline once by gavage); (2) AA-I group (administrated with AA-I 40 mg/kg by gavage once); (3) AA- Group II (AA-II 40 mg/kg was administered by gavage once); (4) AA-IVa group (AA-IVa 40 mg/kg was administered by gavage once).
  • AA-I is highly toxic, and a single intragastric administration can cause the death of many animals. 7 out of 10 animals died, with a mortality rate of 70%; renal pathological examination revealed obvious renal insufficiency. Toxicity, showing diffuse tubular necrosis. AA-II also has obvious toxicity and can cause death of animals, with 1 out of 10 animals dying (mortality rate 10%); renal pathological examination shows patchy renal tubular necrosis, renal tubular swelling, etc. AA-IVa had no obvious toxicity and no animals died; renal pathological examination showed no toxic damage. The results showed that AA-I and AA-II were lethal and nephrotoxic, while AA-IVa had no obvious toxicity and no nephrotoxicity. It shows that the toxicity of AA-IVa is completely different from the toxicity of AA-I and AA-II. The pathological examination results are shown in Figure 5.

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Abstract

本发明提供马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用，其以马兜铃酸IVa及其药学上可接受的盐作为活性组分，一方面可以通过抑制血管通透性从而达到抗组胺作用，另一方面能通过降低组织炎症渗出量从而实现治疗肺炎。

Description

马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用 技术领域
本发明属于医药新用途技术领域，具体涉及马兜铃酸IVa在制备抗组胺药物或治疗肺炎药物中的应用。
背景技术
组胺是一种过敏和炎性介质。许多组织，特别是皮肤、肺和肠粘膜的肥大细胞中含有大量的组胺。当组织受到损伤或发生炎症和过敏反应时，都可释放组胺。组胺有强烈的舒血管作用，并能使毛细血管和微静脉的管壁通透性增加，血浆漏入组织，导致局部组织水肿。临床上很多疾病与组胺释放有关，如皮肤过敏、鼻炎、哮喘、过敏性休克等。
肺炎是由多种病原体(如病毒、细菌、真菌、寄生虫)、放射线、化学、过敏等因素引起。典型性肺炎是由肺炎链球菌等常见细菌引起的大叶性肺炎或支气管肺炎。非典型性肺炎是由支原体、衣原体、军团菌、立克次体、病毒以及其他一些不明微生物引起的肺炎，其中SARS病毒、中东呼吸综合征病毒(MERS)引起的非典型肺炎可引起严重的肺水肿及多脏器功能衰竭。新型冠状病毒患者中少数重型或危重型患者(约20％)的肺部器官会出现病毒增殖，引起肺泡上皮细胞损伤，继而引起肺炎，甚至免疫级联反应和炎症风暴，最终导致严重的非典型双侧间质性肺炎。
急性肺炎症肺部的主要病理变化为急性炎症渗出，严重时可引起肺水肿，急性肺水肿是肺部急性感染和化学性肺损伤(氯气或光气中毒)导致病人死亡的重要原因。
马兜铃酸类是硝基菲羧酸类化合物，以往认为马兜铃酸有肾毒性和致癌性。但马兜铃酸类有很多种类，不同的马兜铃酸毒性差别很大，有毒的马兜铃酸主要为AA-I、AA-II。有研究证明马兜铃酸IVa(AA-IVa)没有明显的毒性，因此其安全性好，有良好的药用用途的前景。经检索，目前尚未发现马兜铃酸IVa在制备抗组胺药物或治疗肺炎药物的相关实验研究。
发明内容
本发明的目的在于提供马兜铃酸IVa的医药新用途，具体地提供了一种以马兜铃酸IVa及其药学上可接受的盐作为活性组分在制备抗组胺药物或治疗肺炎药物中的应用。
为了实现上述目的，本发明采用以下的技术方案：
马兜铃酸IVa在制备抗组胺药物或治疗肺炎药物中的应用。
优选地，所述马兜铃酸IVa的分子式为：C17H11NO8，化学结构式为：
Figure PCTCN2022111333-appb-000001
优选地，所述抗组胺药物的用途，包括用于皮肤过敏、湿疹、皮炎、鼻炎或哮喘。
优选地，所述肺炎包括病原体、放射线、化学、过敏因素引起的肺炎和非典型性肺炎。
进一步优选地，所述病原体包括病毒、细菌、真菌和寄生虫中的至少一种。
进一步优选地，所述非典型性肺炎为SARS、MERS病毒、新型冠状病毒、腺病毒、流感病毒和军团菌中的至少一种引起的。
优选地，所述药物，包含活性成分的马兜铃酸IVa和药学可接受的载体。
优选地，所述载体包括稀释剂、粘合剂、吸收剂、崩解剂、分散剂、湿润剂、助溶剂、缓冲剂和表面活性剂中的至少一种。
优选地，所述药物的给药方式为：口服、舌下、口腔粘膜、静脉内、肌肉内、腹腔内、皮下、经皮、鼻腔和直肠途径中的至少一种。
优选地，所述药物为固体、液体或气体形式。
进一步优选地，所述固体形式为粉末剂、片剂、颗粒剂、丸剂、硬胶囊或软胶囊、乳膏剂、软膏剂、硬膏剂、凝胶剂、糊剂、散剂或贴剂。
进一步优选地，所述液体形式为溶液剂、混悬剂、注射剂、糖浆剂、搽剂、乳剂、酊剂或酏剂。
进一步优选地，所述气体形式为：气雾剂或喷雾剂。
本发明的另一目的在于提供一种抗组胺药物，包含活性成分马兜铃酸IVa或其医学上可接受的盐。
本发明的另一目的还在于提供一种治疗肺炎的药物，包含活性成分马兜铃酸IVa或其医学上可接受的盐。
本发明具有以下有益效果：
本发明所述马兜铃酸IVa能显著抑制组胺诱导的血管通透性增高，表现出显著的抗组胺作用。同时，所述马兜铃酸IVa能够显著降低肺组织炎症渗出量。
通过上述试验可知，本发明所述的马兜铃酸IVa通过抑制血管通透性从而达到抗组胺作用，同时，所述的马兜铃酸IVa通过降低组织炎症渗出量从而实现治疗肺炎。
含马兜铃酸中药具有肾毒性，相比现有技术，本发明活性成分马兜铃酸IVa没有明显的毒性，安全性好，为含马兜铃酸中药的应用提供了新的方向。
附图说明
图1为AA-IVa减轻组胺诱导的血管通透性增高(表现为耳蓝染)。
图2为AA-IVa减轻肺组织炎性渗出(HE病理图片)。
图3为AA-I和AA-II对组胺诱导的血管通透性增高的影响。
图4为AA-I和AA-II对肺组织炎性渗出的影响(HE病理图片)。
图5为AA-I、AA-II和AA-IVa的肾脏毒性比较(肾脏病理图片)。
其中，A为对照组，B为组胺组，C为组胺+AA-IVa/低剂量组，D为组胺+AA-IVa/高剂量组，E为组胺+AA-I组，F为组胺+AA-II组。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚，下面结合具体实施例对本发明的技术方 案做进一步详述。除非另外定义，本文中使用的所有技术和科学术语具有与本发明所属技术领域的普通技术人员通常理解的相同意义。
实施例1：AA-IVa对组胺诱导的小鼠血管通透性增高的影响
1、试验试剂
伊文思蓝(EB)：国药集团化学试剂有限公司，批号：20180125；0.9％氯化钠注射液：辰欣药业股份有限公司，批号：1801022725；甲酰胺：天津市大茂化学试剂厂，批号：20180416；组胺：国药集团化学试剂有限公司，批号20160711；AA-IVa(分子式为：C17H11NO8，分子量为293.27)，源于中药细辛，购自北京赛百草科技有限公司，批号：SH18121005。
2、试验材料
小鼠品系：ICR；性别：雄性。
动物体重：选用23-25g动物，适应性饲养1天后开始给予受试物。来源：北京维通利华实验动物技术有限公司。动物要求：无特殊病原菌。
试验期间动物饲养地点：使用中国中医科学院中药研究所屏障环境动物实验室。
饲养条件：屏障系统，温度20～26℃，相对湿度为40～70％，全新风。采取人工光照，12小时明暗周期。动物饲养于聚碳酸酯小鼠饲养笼，每笼5只小鼠。
饲料：使用标准小鼠颗粒饲料，由北京市科澳协力饲料有限公司提供。饮用水：饮用纯净水，每周更换两次高压灭菌的饮水瓶。
3、试验仪器
动物天平：Sartorius，德国，型号，BSA3202s-CW、BSA224S-CW；
酶标仪：Thermo，美国，型号，VARIOSKAN LUX。
4、试验方法
4.1动物分组
ICR小鼠分为6组，包括阴性对照组(对照)：给予生理盐水灌胃3天，第4天给予生 理盐水尾静脉注射；组胺模型组(组胺)：给予生理盐水灌胃3天，第4天给予组胺5mg/kg尾静脉注射；AA-IVa低剂量治疗组(组胺+AA-IVa/L)：给予AA-IVa 5mg/kg灌胃3天，第4天给予组胺5mg/kg尾静脉注射；AA-IVa高剂量治疗组(组胺+AA-IVa/H)：给予AA-IVa10mg/kg灌胃3天，第4天给予组胺5mg/kg尾静脉注射。
4.2药物配制
AA-IVa：使用0.5wt％CMC-Na配置，浓度分别为0.5mg/ml和0.25mg/ml。组胺：生理盐水配置0.5mg/ml的组胺溶液。
4.3造模方法
(1)原理：组胺可以引起毛细血管和微静脉的管壁通透性增加，血浆漏入组织，导致局部组织水肿。伊文斯兰(EB)是常用于检测血管通透性的指示剂。当小鼠静脉注射组胺与EB混合的溶液后，由于组胺导致血管通透性增高，可以观察到小鼠耳廓血管通透性增高(耳廓染成蓝色)。
(2)组胺诱导耳廓血管通透性增高模型建立的方法：组胺(0.5mg/ml)与0.8％EB溶液以1:1(体积)混合后，根据各只小鼠体重，按20ml/kg体积进行尾静脉注射。给予组胺后观察30min内小鼠的耳廓蓝染情况，拍耳廓蓝染照片。
(3)耳廓血管通透性检测(EB渗出检测)：上述耳廓蓝染观察和拍照后，剪取双耳，剪碎。在室温下用2ml甲酰胺浸泡48h后，用200目不锈钢筛网过滤。取滤液，于610nm处测吸光度(A)。根据伊文思蓝标准曲线，计算伊文思蓝染料渗出量(μg)。
EB渗出降低率(％)＝(组胺组EB渗出平均值—给药组EB渗出平均值)/组胺组EB渗出平均值×100％。
5、统计方法
用SPSS 20.0软件进行统计分析。耳廓组织EB渗出量采用ANOVA程序进行方差分析。数据以均数±标准差
Figure PCTCN2022111333-appb-000002
表示，组胺模型组与对照组比较，各给药组与组胺模型组进行比较，P<0.05为有显著差异。
6、试验结果
组胺为模型组，给予组胺后，小鼠耳廓出现显著的血管通透性增高，耳廓明显蓝染(见图1B)。
不同剂量AA-IVa组动物耳蓝染与组胺模型组相比均明显减轻(见图1C、图D)，EB渗出量较组胺模型组显著降低(P<0.001)，说明AA-IVa有明显的抗组胺作用。
将每组各只小鼠的耳朵泡入甲酰胺溶液中，将EB溶出，再使用酶标仪610nm进行测量，可得到小鼠耳朵中渗出的EB量，体现了血管通透性增高的程度。结果如表1所示，组胺模型组与对照组比较EB渗出量显著增高(与对照组比较P<0.001)；不同剂量AA-IVa组与组胺模型组相比均显著降低(与组胺模型组比较P<0.001)。结果表明，AA-IVa能显著抑制组胺诱导的血管通透性增高，即具有显著的抗组胺作用。
表1不同剂量AA-IVa缓解组胺诱导的耳EB渗出(n＝10)
Figure PCTCN2022111333-appb-000003
注：*代表与对照组比较***P<0.001；#与组胺模型组比较，###P<0.001。
对比例1马兜铃酸I(AA-I)和马兜铃酸II(AA-II)对组胺诱导的小鼠血管通透性的影响
1、试验试剂
伊文思蓝(EB)：国药集团化学试剂有限公司，批号：20180125；0.9％氯化钠注射液：辰欣药业股份有限公司，批号：1801022725；甲酰胺：天津市大茂化学试剂厂，批号：20180416；组胺：国药集团化学试剂有限公司，批号20160711；AA-I(分子式C17H11NO7，分子量：341.27)、AA-Ⅱ(分子式为：C16H9NO6，CAS号：475-80-9，分子量为311.25)，购自北京赛百草科技有限公司。
2、试验材料
同实施例1。
3、试验仪器
同实施例1。
4、试验方法
4.1动物分组
ICR小鼠分为4组，包括(1)阴性对照组(对照)：给予生理盐水灌胃3天，第4天给予生理盐水尾静脉注射；(2)组胺模型组(组胺)：给予生理盐水灌胃3天，第4天给予组胺5mg/kg尾静脉注射；(3)AA-I治疗组(组胺+AA-I)：给予AA-I 10mg/kg灌胃3天，第4天给予组胺5mg/kg尾静脉注射；(4)AA-Ⅱ治疗组(组胺+AA-II)：给予AA-Ⅱ 10mg/kg灌胃3天，第4天给予组胺5mg/kg尾静脉注射。
4.2药物配制
AA-I、AA-Ⅱ配制：采用0.5wt％CMC-Na分别将AA-I和AA-II配成0.5mg/ml。
组胺配制：采用生理盐水配成0.5mg/ml的组胺溶液。
4.3造模方法
同实施例1。
5、统计方法
同实施例1。
6、试验结果
组胺为模型组，给予组胺后，小鼠耳廓出现显著的血管通透性增高，耳廓明显蓝染(见图3B)。
AA-I、AA-Ⅱ组动物耳蓝染与组胺模型组相比无明显差异(见图3E、图3F)，说明AA-I和AA-Ⅱ无明显的抗组胺作用。
将每组各只小鼠的耳朵泡入甲酰胺溶液中，将EB溶出，再使用酶标仪610nm进行测量，可得到小鼠耳朵中渗出的EB量，体现了血管通透性增高的程度。结果如表2所示，组胺组与对照组相比EB渗出量明显升高(P<0.001)；AA-I、AA-Ⅱ组与组胺模型组相比无明显差异，说明AA-I、AA-Ⅱ无显著的抗组胺作用。
表2 AA-I和AA-II对组胺诱导的耳EB渗出无明显改善(n＝10)
Figure PCTCN2022111333-appb-000004
Figure PCTCN2022111333-appb-000005
注：(1)*代表与对照组比较，***P<0.001。(2)AA-I组以及AA-II组与组胺模型组比较均无统计学显著性差异。
实施例2：AA-IVa缓解组胺诱导的肺炎
1、实验方法
(1)模型的建立以及动物分组及给药同实施例1。
(2)肺组织的病理检查：脱椎处死小鼠后，解剖，将肺组织取出。用10％的中性福尔马林将肺组织固定备用。送检组织经甲醛充分固定后，逐级乙醇脱水，二甲苯透明，石蜡包埋，常规制备3μm石蜡切片。分别用HE、Masson、PAS染色，光学显微镜(DP71型，OLYMPUS，放大400倍)检查组织炎症渗出情况。
2、实验结果
肺组织病理结果显示：图2A为正常对照组，显示肺组织肺泡与气管结构正常。图2B为模型对照组，显示肺泡间隔增厚，炎性细胞渗出增多。图2C为组胺+AA-IVa/L组，显示肺泡间隔增厚，炎性细胞渗出增多，但是与模型组相比有明显减轻。图2D为组胺+AA-IVa/H组，显示肺泡间隔增厚，炎性细胞渗出增多，但是与模型组相比有明显减轻。
对比例2AA-I、AA-Ⅱ对组胺诱导肺炎的影响
1、实验方法
(1)模型的建立以及动物分组及给药同对比例1
(2)肺组织的病理检查：同实施例2。
2、实验结果
图4肺组织病理结果显示：图4A为正常对照组，显示肺组织肺泡与气管结构正常。图4B为组胺模型组，显示肺泡间隔增厚，炎性细胞渗出增多，提示肺炎明显。图4E为AA-I治疗组，显示肺泡间隔增厚，炎性细胞渗出明显增多，与组胺模型组相比无明显差异，提示AA-I治疗对组胺诱导的肺炎无明显改善作用。
图4F为AA-Ⅱ组，显示肺泡间隔增厚，炎性细胞渗出增多，与组胺模型组相比无明显差异，提示AA-II治疗对组胺诱导的肺炎无明显改善作用。
实施例3毒性试验
1、试验方法
采用SPF级ICR小鼠，18-22g，雌雄各半。动物来源于北京维通利华实验动物技术有限公司。适应性饲养1天后开始给予受试物。
将小鼠随机分为4组：(1)对照组(灌胃给予生理盐水1次)；(2)AA-I组(灌胃给予AA-I 40mg/kg 1次)；(3)AA-II组(灌胃给予AA-II 40mg/kg 1次)；(4)AA-IVa组(灌胃给予AA-IVa 40mg/kg 1次)。
动物灌胃给药前，均禁食不禁水12h。灌胃给药后，记录灌胃给药后14天内动物死亡情况。存活的动物解剖，取肾脏进行病理检查。
2、结果
如表3结果显示，AA-I具有强烈的毒性，一次灌胃给药就可引起较多的动物死亡，10只动物中死亡了7只，死亡率70％；肾脏病理检查发现有明显的肾脏毒性，呈现弥漫性肾小管坏死。AA-II也有明显的毒性，可引起动物死亡，10只动物中有1只死亡(死亡率10％)；肾脏病理检查可见片状肾小管坏死，肾小管肿胀等。AA-IVa无明显毒性，无动物死亡；肾脏病理检查未见毒性损伤。结果表明，AA-I和AA-II具有致死性和肾毒性，而AA-IVa无明显毒性，无肾脏毒性。说明AA-IVa的毒性与AA-I和AA-II的毒性完全不同。病理检查结果见图5。
表3 AA-IVa与AA-I、AA-II的毒性比较
Figure PCTCN2022111333-appb-000006
需要强调的是，本发明所述的实施例是说明性的，而不是限定性的，因此本发明包括并 不限于具体实施方式中所述的实施例，凡是由本领域技术人员根据本发明的技术方案得出的其他实施方式，同样属于本发明保护的范围。

Claims (9)

  1. 马兜铃酸IVa在制备抗组胺药物或治疗肺炎药物中的应用。
  2. 根据权利要求1所述的应用，其特征在于，所述马兜铃酸IVa的分子式为：C 17H 11NO 8，化学结构式为：
    Figure PCTCN2022111333-appb-100001
  3. 根据权利要求1所述的应用，其特征在于，所述抗组胺药物的用途，包括用于皮肤过敏、湿疹、皮炎、鼻炎或哮喘。
  4. 根据权利要求1所述的应用，其特征在于，所述肺炎，包括病原体、放射线、化学、过敏因素引起的肺炎和非典型性肺炎。
  5. 根据权利要求1所述的应用，其特征在于，所述非典型性肺炎为SARS、MERS病毒、新型冠状病毒、腺病毒、流感病毒和军团菌中的至少一种引起的。
  6. 根据权利要求1所述的应用，其特征在于，所述药物，包含马兜铃酸IVa和药学可接受的载体。
  7. 根据权利要求6所述的应用，其特征在于，所述载体，包括稀释剂、粘合剂、吸收剂、崩解剂、分散剂、湿润剂、助溶剂、缓冲剂和表面活性剂中的至少一种。
  8. 一种抗组胺药物，其特征在于，包含马兜铃酸IVa或其医学上可接受的盐。
  9. 一种治疗肺炎的药物，其特征在于，包含活性成分马兜铃酸IVa或其医学上可接受的盐。
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103387532A (zh) * 2012-05-11 2013-11-13 北京大学 一种抗马兜铃内酰胺FI和马兜铃酸IVa单克隆抗体的制备方法及其应用
CN110194758A (zh) * 2019-04-29 2019-09-03 山东省分析测试中心 一种从关木通中快速分离纯化马兜铃酸类化合物的方法
CN112209948A (zh) * 2020-09-21 2021-01-12 西北农林科技大学 一种马兜铃酸衍生物及应用
CN112237168A (zh) * 2019-07-18 2021-01-19 上海交通大学 用马兜铃酸i或其联合四氯化碳构建小鼠肝癌模型的方法
CN114469930A (zh) * 2022-04-12 2022-05-13 中国中医科学院中药研究所 马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103989670A (zh) * 2013-02-20 2014-08-20 孔徐生 马兜铃酸在制备治疗类风湿性关节炎的药物中的应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103387532A (zh) * 2012-05-11 2013-11-13 北京大学 一种抗马兜铃内酰胺FI和马兜铃酸IVa单克隆抗体的制备方法及其应用
CN110194758A (zh) * 2019-04-29 2019-09-03 山东省分析测试中心 一种从关木通中快速分离纯化马兜铃酸类化合物的方法
CN112237168A (zh) * 2019-07-18 2021-01-19 上海交通大学 用马兜铃酸i或其联合四氯化碳构建小鼠肝癌模型的方法
CN112209948A (zh) * 2020-09-21 2021-01-12 西北农林科技大学 一种马兜铃酸衍生物及应用
CN114469930A (zh) * 2022-04-12 2022-05-13 中国中医科学院中药研究所 马兜铃酸IVa在制备抗组胺或治疗肺炎药物中的应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JING CHEN, SUN BO-HANG, HUANG JIAN, GAO HUI-YUAN, WANG JIAN-QIANG, WU LI-JUN: "Determination of aristolochic acid IVA in the different parts of Asarum heterotropoides Fr.Schmidt var.mandshuricum(Maxim.", JOURNAL OF SHENYANG PHARMACEUTICAL UNIVERSITY, vol. 28, no. 10, 20 October 2011 (2011-10-20), pages 797 - 800, XP093099641, DOI: 10.14066/j.cnki.cn21-1349/r.2011.10.003 *
XIAN ZHONG; TIAN JINGZHUO; ZHANG YUSHI; MENG JING; ZHAO YONG; LI CHUNYING; YI YAN; HAN JIAYIN; LIU SUYAN; WANG LIANMEI; PAN CHEN; : "Study on the potential nephrotoxicity and mutagenicity of aristolochic acid IVa and its mechanism", BIOMEDICINE & PHARMACOTHERAPY, ELSEVIER, FR, vol. 142, 25 August 2021 (2021-08-25), FR , XP086785695, ISSN: 0753-3322, DOI: 10.1016/j.biopha.2021.112081 *

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