CN118078818A - 一种半通道抑制剂在药学上的应用 - Google Patents
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Abstract
本发明提供了一种半通道抑制剂在药学上的应用;其中,所述半通道抑制剂为结构式Ⅰ所示的化合物或其药学上可接受的盐,所述半通道抑制剂用于治疗与连接蛋白半通道活性相关疾病,如抑郁症。
Description
技术领域
本发明属于抗抑郁药物领域,具体涉及一种新型半通道抑制剂及其制备方法和药学上的应用。
背景技术
抑郁障碍为一种最常见及高致残率的精神障碍,抑郁障碍的发病机制涉及多种神经递质系统功能异常、其他脑神经系统如神经免疫系统稳态失衡以及这些神经系统间的相互作用和调节失常;抗抑郁药物是临床上治疗抑郁障碍的主要方法,目前,临床存在多种抗抑郁症药物可用于干预抑郁障碍,但这些传统及新型抗抑郁症药物具有突出不足,包括起效慢,应答率低,耐药,以及不良反应等,具体例如:(1)起效慢,例如多种传统的抗抑郁症药物需要数周及更长时间才能发挥临床效应;(2)治疗效果不佳,主要表现为患者经抗抑郁药治疗无效;(3)靶向性单一,如多数抗抑郁药直接作用于神经递质系统,较少靶向于抑郁障碍密切相关的神经免疫系统。
现存的抗抑郁药主要作用于不同类型的神经递质系统,例如抑郁障碍治疗一线用药的20余种传统抗抑郁药作用于单胺类神经递质系统,如5-羟色胺;美国食品药品监督管理局刚批准用于难治性抑郁的新型快速起效NMDA受体拮抗剂艾司氯胺酮,其作用于谷氨酸能神经递质系统并能影响γ-氨基丁酸GABA能神经系统功能。发现新型有效且安全的抗抑郁药物是当今抑郁障碍研究领域的需要。最近的临床和临床前研究仍在寻找合适的抑郁障碍治疗药物靶点。
发明内容
为了解决上述问题,本发明提供了一种化合物或其药学上可接受的盐在用于制备治疗与连接蛋白半通道相关疾病的药物中的应用;其中,所述化合物具有结构式Ⅰ所示的结构,
根据本发明的具体实施方案,优选地,所述疾病为与抑制连接蛋白半通道活性相关疾病。
根据本发明的具体实施方案,优选地,所述疾病为与抑制连接蛋白半通道活性相关的神经免疫系统功能异常。
根据本发明的具体实施方案,优选地,所述疾病为与神经免疫系统功能异常相关的疾病。
根据本发明的具体实施方案,优选地,所述疾病选自抑郁症、神经炎症。
根据本发明的具体实施方案,优选地,所述化合物或其药学上可接受的盐制备成药物组合物,该药物组合物包含所述的化合物或其药学上可接受的盐作为活性成分,以及一种或多种药学可接受的载体或赋形剂。
根据本发明的具体实施方案,优选地,所述药物组合物的剂型包括混悬剂或胶囊。
根据本发明的具体实施方案,优选地,所述药物组合物剂型为硬胶囊,且单个硬胶囊包含30mg结构式Ⅰ所示的化合物。
根据本发明的具体实施方案,优选地,所述药物组合物通过口服途径或注射途径给药。
根据本发明的具体实施方案,优选地,所述药物组合物的单次口服给药剂量为5mg/kg,所述单次口服给药剂量为所述药物组合物中的结构式Ⅰ所示的化合物的给药剂量。
本发明所提供的化合物以D4为例,是一种新型半通道抑制剂,其作用于一种与抑郁障碍潜在发病机制有关的新型非传统靶点:连接蛋白半通道;小分子化合物D4阻断半通道活性,能减轻与抑郁障碍相关的脑内神经炎症、减弱神经免疫系统异常,如炎症因子表达、星形胶质细胞和小胶质细胞增殖,从而达到缓解抑郁样症状的作用;化合物D4难溶于水,可用于口服,口服后能透过消化道吸收进入血流循环;此外,化合物D4也可注射入受试者,如小鼠;化合物D4通过血脑屏障进入脑后,能特异性抑制脑半通道,通过作用于由星形胶质细胞和小胶质细胞介导的脑内神经炎症反应,化合物D4可防治与抑郁障碍相关的症状,并可避免出现与作用于神经递质系统的谷氨酸、GABA和5-羟色胺等抗抑郁药物有关的药物副作用;另外,化合物D4具有成为快速抗抑郁药物的潜能,其疗效出现于治疗后数日而非数周,并且少次给药可达到长期持续的疗效,作为一种难溶性无毒的有机小分子化合物,化合物D4无明显副作用。在以后的药物研发过程中,化合物D4的这些特性能尽可能减少安全隐患,并能结合多种不同药物递送方法发挥疗效。
本发明提供的半通道抑制剂化合物能特异性作用于脑内半通道蛋白,从而可减轻多种脑系统中与抑郁障碍相关的病理变化,如胶质细胞源性递质系统,神经递质系统,和神经免疫系统。本发明中以D4为例的半通道抑制剂靶向脑内多系统,可能为今后研发更有效的新抗抑郁药物具有重要启示作用。
附图说明
图1为化合物D4的合成路线流程图;
图2为化合物D4的HPLC图;
图3为化合物D4的1H-NMR谱图;
图4为化合物D4的13C-NMR谱图。
具体实施方式
为了对本发明的技术特征、目的和有益效果有更加清楚的理解,现对本发明的技术方案进行以下详细说明,但不能理解为对本发明的可实施范围的限定。
制备例1
本制备例合成了化合物D4,其合成路线如图1所示,由以下步骤制备而得:
(1)制备有机中间体(R)-(2-(4-氯苯)-1,3-二噻烷-2-基)(苯基)甲醇(A905)如图1所示;
(2)由中间体(R)-(2-(4-氯苯)-1,3-二噻烷-2-基)(苯基)甲醇(A905)合成目标产物化合物D4(A907);
化合物D4化学式:C24H16ClNO3;HPLC纯度大于99%,HPLC图如图2所示;
经核磁共振氢谱与核磁共振碳谱验证其结构,如图3和图4所示。图3中氢核磁共振(1H-NMR)指纹图谱显示,低场区7.17-8.4ppm为16个芳香氢信号,其中双峰提示某些氢对称存在。因此,1H-NMR光谱指纹图谱与化合物D4的分子式C24H16ClNO3中所含的16个氢原子和结构式中含有的4个苯环信息一致。图4中固体13C核磁共振谱(13C NMR)显示共有20个谱峰。13CNMR谱中对称分布的4个碳相应的谱峰重叠。13C NMR谱192.227ppm处为羰基的特征峰,164.494ppm处为酯基的特征峰,121.2887-147.83ppm区域的17个峰为苯环中碳的特征峰,78.865ppm处为碳卤(氯)键的特征峰。因此,13C NMR谱中的24个碳原子与化合物D4的分子式和结构式一致。
除特别说明外,本发明测试例所采用的实验动物均为C57BL/6成年小鼠(3-4月龄),体重29.1±0.6g;本发明测试例所采用的化合物D4均以混悬液形式经口服给药,该混悬液采用含10%乙醇的灭菌生理盐水作为溶剂制成,且每毫升含1毫克化合物D4。
测试例1
本测试例考察了在急性炎症模型中,化合物D4对连接蛋白半通道的抑制作用。
成年小鼠经腹腔注射单剂量高浓度LPS(1mg/kg),以构建急性炎症模型。LPS注射24小时后,麻醉成年小鼠并获取海马区脑片,采用CBF荧光染料摄取方法检测连接蛋白半通道开放程度;
小鼠腹腔注射单剂量LPS(1mg/kg)24小时后,经腹腔注射氯胺酮(100mg/kg)/甲苯噻嗪(10mg/kg)混合液麻醉后,经当天配制预通95%O2和5%CO2混合气饱和的冰冷人工脑脊液(artificial cerebrospinal fluid,ACSF;19mM NaCl,2.5mM KCl,2.5mM CaCl2,1.3mMMgSO4,1mM NaH2PO4,26.2mM NaHCO3,and 22mM葡萄糖)灌注后,于冰上在冰冷人工脑脊液中快速取脑,使用振动切片机(VT1200S,Leica),于当天配制预通95%O2和5%CO2混合气饱和的冰冷ACSF中,制备300μm厚度含海马的冠状脑片;随后将脑片移至预通95%O2和5%CO2混合气1h的ACSF中,LPS+空白组(Vehicle)为含10%乙醇的ACSF,LPS+D4组为含D4(0.5mM)和10%乙醇的ACSF。室温下孵育20分钟后,脑片转移至含预通95%O2和5%CO2混合气1h的含CBF(100μM)的ACSF染液中,室温孵育20分钟;孵育结束后,脑片用ACSF漂洗3次,每次漂洗时间为5分钟;漂洗结束后,将海马脑片移至4%PFA中于4℃固定过夜;次日,将海马脑片转移至30%蔗糖溶液中于4℃脱水处理36小时;脱水完成后,剥取海马,包埋后采用冰冻切片机(HM525 NX,Thermo Fisher Scientific)获取15μm厚度的海马切片;切片染色后,于共聚焦显微镜(LSM880,ZEISS)下观察拍照。采用Image J图像分析软件,测量海马齿状回(Dentategyrus,DG)的CBF阳性面积用于分析脑片海马齿状回区的CBF摄取量。
体外实验结果见表1,其中,正常对照组为小鼠经腹腔注射灭菌生理盐水,且脑片于预通95%O2和5%CO2混合气的ACSF中孵育;结果表明,经单剂量高浓度LPS注射的小鼠的海马区脑片中连接蛋白半通道开放显著增加(LPS+空白组),而小鼠经D4预处理后(LPS+D4组)能明显抑制LPS诱导的海马区脑片的连接蛋白半通道开放(每组n=4只小鼠)。
表1D4体外预处理对LPS诱导小鼠海马区脑片中连接蛋白半通道活性的影响
注:单因素方差分析最小显著性差异法多重比较(One-way ANOVA followed byLSD post hoc test)。与正常对照组比较,***p<0.001;与LPS+空白组比较,##p<0.01。
测试例2
本测试例考察了在慢性炎症模型中,化合物D4对连接蛋白半通道的抑制作用。
首先,为检测LPS诱导的慢性炎症对脑内半通道功能的影响,成年小鼠每日腹腔注射单次低剂量LPS(0.75mg/kg),为期一周,从而构建慢性炎症模型,作为LPS组;生理盐水对照组为小鼠每日腹腔注射等体积的灭菌生理盐水;小鼠经最后一次腹腔注射LPS恢复24小时后,在第9天根据测试例1中方法,麻醉后,快速获取含海马的脑片,制备300μm厚度含海马的冠状脑片;随后将脑片移至预通95%O2和5%CO2混合气1h的ACSF中,室温下孵育20分钟;孵育结束后,将脑片转移至含预通95%O2和5%CO2混合气1h的含CBF(100μM)的ACSF染液中,室温孵育20分钟;染色结束后,按照测试例1中方法漂洗脑片及后续处理;切片染色后,拍照测量CBF阳性面积,分析脑片海马齿状回区的CBF摄取量;荧光染料摄取实验结果表明,与生理盐水对照组(n=3只小鼠)相比,LPS组(n=7只小鼠)的CBF荧光染料摄取量显著增加,表明LPS诱导的神经炎症能增强脑内半通道的功能,结果见表2。
表2 LPS慢性炎性抑郁小鼠模型中海马脑片半通道活性的变化
注:曼-惠特尼U检验(Mann–Whitney U test)。与生理盐水对照组比较,***p<0.001。
接着,为检测化合物D4是否影响LPS诱导的慢性炎症中炎症因子水平,成年小鼠连续每日腹腔注射单次低剂量LPS(0.75mg/kg),为期一周;在每次LPS注射后(5min内),小鼠立即经口服给予化合物D4(5.0mg/kg)处理,作为LPS+D4组;LPS+空白组(Vehicle)为小鼠经腹腔注射较低剂量LPS(0.75mg/kg)后,口服给予等体积不添加D4的含10%乙醇的灭菌盐水;生理盐水对照组为小鼠经腹腔注射等体积的灭菌生理盐水后,口服给予等体积不添加D4的含10%乙醇的灭菌盐水;为检测化合物D4对LPS诱导慢性炎症的影响,小鼠经最后一次LPS注射和灌胃给药处理4小时后,收集全血和海马组织,分别用于提取血浆和海马蛋白样品,采用酶联免疫法(ELISA)测定血浆和脑内海马中炎症因子IL-1β水平。由表3可见,与生理盐水对照组相比较,LPS+空白组小鼠血浆和海马中IL-1β水平均有显著升高;LPS+D4组小鼠血浆和海马中IL-1β水平明显降低(每组中n=3只小鼠),提示化合物D4能改善LPS诱导的炎症反应。
表3 D4口服处理对LPS诱导慢性炎症中炎症因子的影响
注:单因素方差分析最小显著性差异法多重比较(One-way ANOVA followed byLSD post hoc test)。与生理盐水对照组比较,**p<0.01,***p<0.001;与LPS+空白组比较,##p<0.01,###p<0.001。
为进一步检测化合物D4是否影响LPS诱导的慢性炎症对脑内半通道功能异常,成年小鼠连续每日腹腔注射单次低剂量LPS(0.75mg/kg),为期一周;在每次LPS注射后(5min内),小鼠立即经口服给予不同剂量的化合物D4(0.5mg/kg、5.0mg/kg)处理,分别作为LPS+D4低剂量组(0.5mg/kg)、LPS+D4高剂量组(5.0mg/kg);LPS+空白组(Vehicle)为小鼠经腹腔注射较低剂量LPS(0.75mg/kg)后,口服给予等体积不添加D4的含10%乙醇的灭菌盐水;小鼠经最后一次LPS注射和给药处理恢复一天后,在第9天根据测试例1中方法,麻醉后,快速获取含海马的脑片,制备300μm厚度含海马的冠状脑片;随后将脑片移至预通95%O2和5%CO2混合气1h的ACSF中,室温下孵育20分钟。孵育结束后,将脑片转移至含预通95%O2和5%CO2混合气1h的含CBF(100μM)的ACSF染液中,室温孵育20分钟;染色结束后,按照测试例1中方法漂洗脑片及后续处理;切片染色后,拍照测量CBF阳性面积,分析脑片海马齿状回区的CBF摄取量。
荧光染料摄取实验结果见表4,结果表明,低剂量D4口服给药组(0.5mg/kg)能减少连接蛋白半通道的激活(LPS+D4低剂量组);此外,高剂量D4口服给药组(5.0mg/kg)对LPS诱导的连接蛋白半通道激活具有更强的抑制效果(LPS+D4高剂量组)。由此,这些实验结果证实D4可作为一种特异性新型半通道抑制剂(LPS+空白组:n=7只小鼠;LPS+D4低剂量组:n=7只小鼠;LPS+D4高剂量组:n=4只小鼠)。
表4 D4口服处理对LPS诱导小鼠海马脑片中半通道活性的影响
注:克鲁斯卡尔-沃利斯检验(Kruskal–Wallis test with Bonferronicorrection)。与LPS+空白组比较,*p<0.05,***p<0.001。
测试例3
本测试例考察了化合物D4在小鼠抑郁障碍模型中的效果。
为检测D4的行为学疗效,以下使用了两种经典的抑郁障碍小鼠模型。
首先使用了LPS慢性炎症的抑郁症模型测试化合物D4是否具有抗抑郁样作用。此模型中,成年小鼠进行了连续一周的每日单剂量低浓度LPS(0.75mg/kg)腹腔注射;每次LPS注射后,小鼠立即经口服给药给予单次的适当剂量的D4(5mg/kg)。LPS注射和给药处理在上午10:00-12:00完成。LPS+空白组为每次LPS注射后,小鼠立即经口服给药给予单次的含10%乙醇的灭菌生理盐水溶液;正常对照组为小鼠连续一周腹腔注射等体积的灭菌生理盐水,灌胃是使用的是含10%乙醇的灭菌生理盐水溶液。连续一周造模和给药结束后,小鼠恢复一天后进行行为学实验,包括旷场实验(Open field test,OFT)、悬尾实验(Tailsuspension test,TST)、强迫游泳实验(Forced swimming test,FST)和蔗糖偏好实验(Sucrose preference test,SPT)。旷场实验在白箱(50cm×50cm×50cm,400Lux)中进行,测试时间为10分钟,记录小鼠的中心区(25cm×25cm)停留时间和总距离。悬尾实验和强迫游泳实验的测试时间为6分钟,记录小鼠的绝对不动时间。蔗糖偏好实验前准备包括蔗糖适应(24小时)和饮水剥夺(36小时),测试当天记录小鼠1小时内饮用1%蔗糖水或正常饮用水的总量。参照汉密尔顿抑郁量表(Hamilton Depression Scale,HAMD)评估抑郁严重程度的主要抑郁症状,如焦虑(Anxiety)、绝望感(Hopelessness)和快感缺失(Anhedonia)等;为评价小鼠的抑郁样水平,综合行为学实验中旷场实验的中心区停留时间、悬尾实验中的绝对不动时间、强迫游泳实验中的绝对不动时间和蔗糖偏好指数为指标,进行Z标准化(标准分数,standard score或Z-score),表示小鼠的抑郁样行为。
进行Z分数标准化时,首先由每只小鼠与生理盐水对照组(正常对照组)的差除以生理盐水对照组的标准差得出各行为学实验中的Z分数,包括旷场实验中周边区域的停留时间Z分数记为ZOFT,检测小鼠的焦虑样水平,分数越高表示小鼠的焦虑样水平越高;悬尾实验和强迫游泳实验中不动时间Z分数分别记为ZTST和ZFST,分数越高表示小鼠的行为绝望程度越高;蔗糖偏好实验中普通正常饮用水Z分数记为ZSPT,分数越高表示小鼠的快感缺失程度越高。抑郁样z分数为ZOFT、ZTST、ZFST和ZSPT的平均值,分数越高表示小鼠的抑郁样水平越高。行为学检测表明,LPS能显著增加小鼠的抑郁样行为(LPS+空白组);与LPS空白组和正常对照组相比,口服给予化合物D4(LPS+D4组)能显著预防LPS诱导的小鼠抑郁样行为(正常对照组n=15只小鼠,10雄5雌;LPS+空白组n=12只小鼠,9雄3雌;LPS+D4组n=14只小鼠,9雄5雌),结果见表5。
表5 D4口服处理对LPS诱发慢性炎症导致小鼠的抑郁样行为的影响
注:单因素方差分析最小显著性差异法多重比较(One-way ANOVA followed byLSD post hoc test)。与正常对照组比较,*p<0.05,***p<0.001;与LPS+空白组比较,###p<0.001。
为进一步验证化合物D4的抗抑郁样作用,小鼠进行每日6小时的束缚处理,为期5周,束缚时间为10:00-16:00。慢性束缚造模结束后,小鼠恢复一天后进行一系列行为学实验,包括旷场实验、悬尾实验、强迫游泳实验和蔗糖偏好实验。在每种行为学检测前16-18小时,小鼠给予对照(含10%乙醇的灭菌生理盐水溶液,作为CRS+空白组)或单剂量D4(5mg/kg)的口服用药处理。正常对照组不经束缚应激处理;按照上述Z标准化方法指示小鼠的抑郁样行为,从而评价D4的治疗效果。行为学检测结果见表6,结果表明,慢性束缚应激小鼠的抑郁样水平明显增加,而D4口服给药显著改善慢性束缚应激(CRS)导致的小鼠抑郁样行为(正常对照组:n=15只小鼠,10雄5雌;CRS+空白组:n=15只小鼠,5雄10雌;CRS+D4组:n=14只小鼠,5雄10雌)。综上所述,D4具有良好的抗抑郁样疗效。
表6 D4口服处理对慢性束缚应激CRS导致小鼠的抑郁样行为的影响
注:单因素方差分析最小显著性差异法多重比较(One-way ANOVA followed byLSD post hoc test)。与正常对照组比较,**p<0.01;与CRS+空白组比较,###p<0.001。
测试例4
本测试例考察了化合物D4在小鼠中发挥抗抑郁样效果的机制。
星形胶质细胞含有丰富的连接蛋白表达;化合物D4的主要作用靶点为连接蛋白的半通道,为检测D4对胶质细胞的影响,使用免疫荧光染色方法,以胶原纤维酸性蛋白(GFAP)为标记物,观察化合物D4对两种抑郁模型小鼠中海马GFAP阳性细胞的变化。
在LPS慢性炎症抑郁模型小鼠中,为检测D4对LPS诱导的抑郁样小鼠海马区星形胶质细胞的影响,按照测试例3中方法,构建LPS慢性炎症抑郁模型,成年小鼠进行连续一周的每日单剂量低浓度LPS(0.75mg/kg)腹腔注射,每次LPS注射后,小鼠立即经口服给药给予单次的适当剂量的D4(5mg/kg),作为LPS+D4组,LPS+空白组小鼠则给予等体积的含10%乙醇的灭菌生理盐水。造模和给药结束后,小鼠恢复一天用于行为学检测;最后的蔗糖偏好实验结束后,小鼠经深度麻醉,以4%多聚甲醛灌注固定,在4%多聚甲醛于4℃中固定过夜后,于30%蔗糖溶液中脱水三天,随后制备厚度为30μm的冰冻脑切片,采用免疫荧光染色观察并分析GFAP阳性星形胶质细胞的数量。免疫荧光染色结果显示,与LPS+空白组相比,LPS+D4组小鼠海马星形胶质细胞的数量显著减少,表明5mg/kg化合物D4能够显著抑制LPS慢性炎症介导的抑郁样小鼠海马中星形胶质细胞变化,结果见表7。
表7 D4口服处理对LPS慢性炎症抑郁样小鼠海马星形胶质细胞的影响
注:曼-惠特尼U检验(Mann-Whitney U test)。与LPS+空白组比较,***p<0.001。
为进一步验证化合物D4对胶质细胞的影响,检测化合物D4对CRS模型小鼠中海马胶质细胞数量的影响,以GFAP为标记物观察星形胶质细胞数量的变化,以IBA1为标记物观察小胶质细胞数量的变化;小鼠进行每日6小时的束缚处理,为期5周,束缚时间为10:00-16:00;慢性束缚造模结束后,小鼠恢复一天后进行一系列行为学实验,包括旷场实验、悬尾实验、强迫游泳实验和蔗糖偏好实验,在每种行为学检测前16-18小时,小鼠给予对照(含10%乙醇的灭菌生理盐水溶液,作为CRS+空白组)或单剂量D4(5mg/kg)的口服用药处理。最后一次行为实验结束后,按照上述方法灌注固定并制备30μm的冰冻脑切片,采用免疫荧光染色观察并分析GFAP阳性星形胶质细胞的数量和IBA1阳性小胶质细胞的数量。与CRS+空白组相比较,CRS+D4组小鼠的海马星形胶质细胞(各组n=5只小鼠)和小胶质细胞(各组n=5只小鼠)的数量显著降低,结果分别见表8和表9。
表8 D4口服处理对CRS抑郁样小鼠海马星形胶质细胞的影响
注:独立样本t检验(Unpaired Student's t-test)。与LPS+空白组比较,*p<0.05。
表9 D4口服处理对CRS抑郁样小鼠海马小胶质细胞的影响
注:曼-惠特尼U检验(Mann-Whitney U test)。与LPS+空白组比较,*p<0.05。
本发明提供的半通道抑制剂化合物能特异性作用于脑内半通道蛋白,可用于缓解抑郁样症状,同时,小鼠经D4给药后,其活动和躯体反应,如笼内的运动、体重和体温,均未发现化合物D4处理的小鼠有异常变化,因此,化合物D4作为新型抗抑郁药具有很大应用前景。
Claims (10)
1.一种化合物或其药学上可接受的盐在用于制备治疗与连接蛋白半通道相关疾病的药物中的应用;其中,所述化合物具有结构式Ⅰ所示的结构,
2.根据权利要求1所述的应用,其中,所述疾病为与抑制连接蛋白半通道活性相关疾病。
3.根据权利要求1所述的应用,其中,所述疾病为与抑制连接蛋白半通道活性相关的神经免疫系统功能异常。
4.根据权利要求1所述的应用,其中,所述疾病为与神经免疫系统功能异常相关的疾病。
5.根据权利要求1所述的应用,其中,所述疾病选自抑郁症、神经炎症。
6.根据权利要求1-5任一项所述的应用,其中,所述化合物或其药学上可接受的盐制备成药物组合物,该药物组合物包含所述的化合物或其药学上可接受的盐作为活性成分,以及一种或多种药学可接受的载体或赋形剂。
7.根据权利要求6所述的应用,其中,所述药物组合物的剂型包括混悬剂或胶囊。
8.根据权利要求6所述的应用,其中,所述药物组合物剂型为硬胶囊,且单个硬胶囊包含30mg结构式Ⅰ所示的化合物。
9.根据权利要求6所述的应用,其中,所述药物组合物通过口服途径或注射途径给药。
10.根据权利要求6所述的应用,其中,所述药物组合物的单次口服给药剂量为5mg/kg,所述单次口服给药剂量为所述药物组合物中的结构式Ⅰ所示的化合物的给药剂量。
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