CN114886913A - 香菇多糖在治疗新型冠状病毒感染中的应用 - Google Patents
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Abstract
本发明涉及医药技术领域，关于香菇多糖在在治疗新型冠状病毒感染中的应用。所述香菇多糖在治疗新型冠状病毒感染中的应用，是以香菇多糖为唯一的有效成分，或包含香菇多糖在内的复合成分，通过滴鼻或鼻腔喷雾途径接种，治疗新型冠状病毒感染的制剂。将新型冠状病毒滴鼻感染叙利亚金黄地鼠，1天以后每天给予滴鼻香菇多糖，结果显示，香菇多糖滴鼻可有效降低地鼠上呼吸道中的病毒载量，减轻地鼠体重降低幅度，并加快地鼠体重恢复，具有应用前景。
Description
技术领域
本发明属于医药技术领域，具体涉及香菇多糖在治疗新型冠状病毒感染中的应用。
背景技术
新型冠状病毒(SARS-CoV-2)，下文中简称为新冠病毒，具有极高的传染性和快速的人间传播能力，可导致部分感染者出现病毒性肺炎(Coronavirus Disease 2019，COVID-19)。这些突变株的共同特征是具备显著的免疫逃逸能力，能突破人群因先前感染病毒或接种疫苗而建立的免疫保护屏障。尤其是2021年11月在南非首次被发现的奥密克戎突变株，具有极强的传染性、传播力和免疫逃逸能力，迅速在全球范围内取代先前流行的主要突变株-德尔塔突变株。
目前针对新冠病毒感染的药物包括针对病毒蛋白酶、RNA聚合酶的小分子抑制剂，以及病毒中和抗体。这些药物虽然有效，但价格较贵，市场供应较为紧张。针对一些局限于上呼吸道的轻症感染，使用这类药物并不必要。但是，如果有廉价且有效的药物可抑制上呼吸道局部新冠病毒的复制，不仅能缓解症状，而且可减轻感染者作为传染源传播病毒的危害。
香菇多糖(Lentinan，LNT)是从优质香菇子实体中提取的有效活性成分，是一种免疫调节剂，尤其是可有效增强T细胞活性。临床与药理研究表明，香菇多糖具有抗病毒、抗肿瘤、调节免疫功能和刺激干扰素形成等作用。其活性成分是具有分支的β-(1-3)-D-葡聚糖，主链由β-(1-3)-连接的葡萄糖基组成，沿主链随机分布着由β-(1-6)连接的葡萄糖基，呈梳状结构。香菇多糖注射液和口服香菇多糖片在包括我国在内的多个国家被临床用于慢性肝炎和肿瘤的辅助治疗，可有效增强患者的抗病毒和抗肿瘤免疫应答，安全性好。
发明内容
本发明旨在提供一种经呼吸道粘膜给药治疗新冠病毒感染的药物。
本发明将新冠病毒滴鼻感染叙利亚金黄地鼠，1天以后每天给予滴鼻香菇多糖，结果显示香菇多糖滴鼻可有效降低地鼠上呼吸道中的病毒载量，减轻地鼠体重降低幅度，并加快其体重恢复，具有应用前景。
本发明提供了香菇多糖在治疗新型冠状病毒感染中的应用。
此外，本发明提供的应用，其特征在于：香菇多糖作为一种有效的免疫调节剂，通过滴鼻或鼻腔喷雾给药，可抑制呼吸道上皮细胞中新冠病毒的复制，促进呼吸道上皮细胞中新冠病毒的清除。
此外，本发明提供的应用，其特征在于：香菇多糖作为一种有效的免疫调节剂，通过滴鼻或鼻腔喷雾给药，可减轻新冠病毒感染引起的疾病症状，缩短疾病时间。
1.此外，本发明提供的应用，，其特征在于：所述香菇多糖应用于治疗新型冠状病毒感染的给药途径包括滴鼻、鼻腔喷雾。
附图说明
图1.新冠病毒攻击后第3、5天叙利亚金黄地鼠鼻甲中新冠病毒N基因相对水平
在病毒攻击后第3和第5天，分别处死感染对照组和香菇多糖滴鼻给药组3 只地鼠，取鼻甲，研磨，抽提组织细胞RNA，用逆转录-定量PCR检测新冠病毒N基因相对表达水平。
图2.新冠病毒感染后叙利亚金黄地鼠的体重变化
空白对照组(未感染病毒，也未滴鼻溶剂或香菇多糖溶液)、感染对照组、香菇多糖滴鼻给药组，每组各6只地鼠在病毒攻击以后每天的体重变化。
具体实施方式
为了更清楚地说明本发明，下面结合优选实施例对本发明做进一步的说明。本领域技术人员应当理解，下面所具体描述的内容是说明性的而非限制性的，不应以此限制本发明的保护范围。
本发明实施例所用的伊沙佐米可以通过市售方式购买获得。
一、病毒、实验动物、药物、试剂
1.病毒：SARS-CoV-2病毒由海军军医大学生物医学防护教研室从COVID-19患者鼻咽拭子样本中分离培养出，其基因序列参见GenBank Accession No.MT622319，用Vero E6细胞培养病毒。培养的病毒感染VeroE6细胞，18 小时以后用免疫荧光染色检测病毒的滴度(focus forming units,FFU)，即每毫升病毒液中感染性病毒颗粒的数量。涉及病毒感染的实验操作均在海军军医大学P3实验室中进行。
2.叙利亚金黄，雄性、8周龄，购自北京维通利华实验动物技术有限公司。
3.香菇多糖(LNT)，购自MCE(MedChemExpress)公司，目录号：HY-N6653，用二甲基亚砜溶解为储存液。
4.RNA抽提试剂Trizol LS购自Thermo Fisher，目录号：10296010。
5.新冠病毒核酸荧光定量PCR检测试剂，购自上海伯杰生物科技有限公司。
二、实验方法：
共24只叙利亚金黄，以滴鼻方式感染新冠病毒，先用异氟烷吸入方式麻醉动物，随后滴鼻攻毒，剂量1.8*108FFU，体积50微升。从病毒攻击当天(滴鼻病毒前)开始称量地鼠体重，此后每天称量体重。在病毒攻击后第1天(攻毒 24小时以后)，将地鼠随机分为2组，每组12只。一组为感染对照组，滴鼻溶剂，为含1％二甲基亚砜的磷酸盐缓冲液，双侧鼻孔滴鼻，25微升/每侧鼻孔；另一组为香菇多糖滴鼻给药组，双侧鼻孔滴鼻，25微升/每侧鼻孔，共含LNT 200 微克。另6只地鼠作为空白对照组，不感染病毒，也不予以滴鼻溶剂或香菇多糖溶液。
滴鼻后第3天和第5天，感染对照组和香菇多糖滴鼻给药组每组取3只地鼠，以异氟烷麻醉后处死，剪切取鼻甲，用匀浆器研磨，用Trizol LS试剂抽提组织细胞RNA，用逆转录实时荧光定量技术检测新冠病毒核衣壳N基因核酸含量，计算各组地鼠鼻甲中病毒核酸的相对水平。两组其余6只地鼠继续观察体重变化，至第14天。
各组在病毒攻击后第3天和第5天的鼻甲中新冠病毒N基因的相对水平如图1所示：将阴性对照组3只地鼠第3天N基因平均水平设置为1，则第5天相对水平为0.974；LNT滴鼻组3只地鼠第3天N基因相对水平为0.445，第5 天为0.228。在第3天和第5天，LNT滴鼻组的N基因相对水平都显著低于阴性对照组(p＜0.001,t检验)。
地鼠体重变化如图2所示：阴性对照组6只地鼠在病毒攻击以后体重持续下降，最大降低幅度达15.4％，至第7天以后开始回升；而LNT滴鼻组，体重下降幅度相对较低，最大降低幅度为8.9％，且提前2天开始回升。
以上实验结果表明，新冠病毒感染地鼠以后，对地鼠用滴鼻途径给药香菇多糖，可有效抑制地鼠鼻甲中的病毒复制，并且减轻地鼠体重降低幅度，缩短发地鼠病时间，发挥治疗新冠病毒感染的效应。
以上所述仅是本发明的较佳实施例而已，并非对本发明作任何形式上的限制，虽然本发明已以较佳实施例揭露如上，然而并非用以限定本发明，任何熟悉本专利的技术人员在不脱离本发明技术方案范围内，当可利用上述提示的技术内容作出些许更动或修饰为等同变化的等效实施例，但凡是未脱离本发明技术方案的内容，依据本发明的技术实质对以上实施例所作的任何简单修改、等同变化与修饰，均仍属于本发明方案的范围内。本发明要求保护范围由所附的权利要求书及其等同物界定。
Claims (4)
1.香菇多糖在治疗新型冠状病毒感染中的应用。
2.根据权利要求1所述的应用，其特征在于：香菇多糖作为一种有效的免疫调节剂，通过滴鼻或鼻腔喷雾给药，可抑制呼吸道上皮细胞中新冠病毒的复制，促进呼吸道上皮细胞中新冠病毒的清除。
3.根据权利要求1所述的应用，其特征在于：香菇多糖作为一种有效的免疫调节剂，通过滴鼻或鼻腔喷雾给药，可减轻新冠病毒感染引起的疾病症状，缩短疾病时间。
4.根据权利要求1-3任一所述的应用，其特征在于：所述香菇多糖应用于治疗新型冠状病毒感染的给药途径包括滴鼻、鼻腔喷雾。
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| PCT/CN2022/101513 WO2023201882A1 (zh) | 2022-04-20 | 2022-06-27 | 香菇多糖在治疗新型冠状病毒感染中的应用 |
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| US20210393681A1 (en) * | 2020-06-22 | 2021-12-23 | Therapeutic Solutions International, Inc. | Treatment of SARS-CoV-2 with Dendritic Cells for Innate and/or Adaptive Immunity |
| US20220054601A1 (en) * | 2020-08-18 | 2022-02-24 | Morehouse School Of Medicine | Methods and compositions for treating coronavirus infections |
| CN114344457A (zh) * | 2021-11-13 | 2022-04-15 | 暨南大学 | 新型冠状病毒蛋白质抗原纳米疫苗及其制备方法和应用 |
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| US20230233596A1 (en) * | 2020-06-16 | 2023-07-27 | Gn Corporation Co Ltd | Beta-glucan for immuno-enhancement and/or immuno-balancing, and for adjuvant use |
| CN111588732A (zh) * | 2020-06-18 | 2020-08-28 | 大连工业大学 | 岩藻多糖在抗新型冠状病毒中的应用 |
| CN111803515A (zh) * | 2020-07-23 | 2020-10-23 | 广东工业大学 | 海藻多糖及其衍生物在制备预防和/或治疗新型冠状病毒感染药物中的应用 |
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| US20210393681A1 (en) * | 2020-06-22 | 2021-12-23 | Therapeutic Solutions International, Inc. | Treatment of SARS-CoV-2 with Dendritic Cells for Innate and/or Adaptive Immunity |
| US20220054601A1 (en) * | 2020-08-18 | 2022-02-24 | Morehouse School Of Medicine | Methods and compositions for treating coronavirus infections |
| CN114344457A (zh) * | 2021-11-13 | 2022-04-15 | 暨南大学 | 新型冠状病毒蛋白质抗原纳米疫苗及其制备方法和应用 |
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| EMMA J. MURPHY: "β-Glucan extracts from the same edible shiitake mushroom Lentinus edodes produce differential in-vitro immunomodulatory and pulmonary cytoprotective effects— Implications for coronavirus disease (COVID-19) immunotherapies", SCIENCE OF THE TOTAL ENVIRONMENT, vol. 732, pages 1 - 10 * |
| SECHUL CHUN: "Antioxidant Activity of Mushroom Extracts/Polysaccharides—Their Antiviral Properties and Plausible AntiCOVID-19 Properties", ANTIOXIDANTS, vol. 10, no. 12, pages 1 - 30 * |
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