CN111068054A - 以csf1r作为药物靶点治疗肿瘤的药剂及其制备方法 - Google Patents
以csf1r作为药物靶点治疗肿瘤的药剂及其制备方法 Download PDFInfo
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Abstract
本发明属于生物医药领域,主要是一种以CSF1R作为药物靶点治疗肿瘤的药剂及其制备方法,本发明涉及一个新的朗格汉斯细胞瘤或肿瘤相关巨噬细胞治疗位点,具体涉及以CSF1R作为靶点治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞,以CSF1R为靶点的Ki 20227口服油剂和CSF1R抑制性单抗溶剂,包括使用它们制备药物作为靶向治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞的应用。CSF1R药物靶点及其拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂在制备治疗朗格汉斯细胞瘤或抑制肿瘤相关巨噬细胞来治疗恶性肿瘤的药物中有着很大的临床运用价值,为治疗朗格汉斯细胞瘤或抑制肿瘤相关巨噬细胞来治疗恶性肿瘤提供了新的方法和药物及制备方法。
Description
技术领域
本发明属于生物医药领域,主要是一种以CSF1R作为药物靶点治疗肿瘤的药剂及其制备方法。
背景技术
恶性肿瘤是严重危害人类生命健康的疾病,我国每年癌症发病人数约380.4万人,而每年因癌症死亡的人数约229.6万人。世界卫生组织2012年《世界癌症报告》指出中国新诊断癌症病例为380.4万,占到全球总数的21.8%;而年死亡人数229.6万,占到全球癌症年死亡人数的24.0%。更为严重的是这些数据正以惊人的速度逐年增长。因此,探索癌症发生的分子机制研究及寻找其有效治疗手段对人类健康具有重大意义。
肿瘤相关巨噬细胞(Tumor-associated macrophages,TAMs)是属于巨噬细胞细胞系的一种细胞,发现于肿瘤组织附近。TAMs由淋巴循环中的单核细胞或组织中残留的巨噬细胞衍生而来,是浸润许多种肿瘤基质的白细胞的主要类型。尽管存在一些争议,但大部分的证据显示TAMs有着促进肿瘤的表型。TAMs影响了肿瘤细胞生理病理的许多方面,包括肿瘤细胞增殖、肿瘤血管发生、侵袭性和恶性转移、免疫抑制以及药物抵抗性。在许多肿瘤类型中,TAMs浸润水平已显示出重要的预后价值。TAMs与乳腺癌,卵巢癌,神经胶质瘤和淋巴瘤的预后不良有关;在结肠癌和胃癌中预后较好,在肺癌和前列腺癌中预后差。
据报道,已经开发出CSF1R抑制剂作为减少肿瘤微环境中TAMs存在的潜在途径。在临床前模型中已经测试了其他增强肿瘤对化学疗法反应的方法,包括阻止巨噬细胞募集到肿瘤部位,重新极化TAMs和促进TAMs活化。靶向TAM的其他挑战包括确定联合疗法是靶向耗竭还是复极化,以及针对哪种肿瘤类型以及TAM靶向治疗在哪种肿瘤阶段有效。通过药物治疗将TAMs从M2型重新极化为M1型已显示出控制肿瘤生长的能力,包括与检查点抑制剂疗法联合使用。
朗格汉斯细胞(Langerhans cells)是一种遍布全身的免疫细胞,特别在皮肤,淋巴结,脾脏,肺,肝脏和骨髓当中存在,其本身起到调节免疫系统的作用。主要存在于皮肤,皮肤的树突细胞(抗原呈递免疫细胞),并含有称为Birbeck颗粒的细胞器。
在朗格汉斯细胞组织瘤(Langerhans cell histiocytosis,LCH)中,过量的未成熟朗格汉斯细胞聚集形成称为肉芽瘤(granulomas)的肿瘤。发病率为1/100000~9/100000,LCH的患病率在白人中高于其他种族的人群,在男性中比在女性中更高,男女比例为2:1。LCH影响从新生儿期到成年期的患者,在0-15岁的儿童中更常见(据报道,每百万人口约4例)。骨是最常受影响的器官(80%的病例),其次是皮肤(35%的病例),然后是脑垂体(25%的病例)。病因未知,可能与基因突变与遗传有关。朗格汉斯细胞增殖可以由病毒感染,细胞间通讯缺陷(T细胞—巨噬细胞相互作用)和由肿瘤坏死因子,IL-11和白血病抑制因子介导的细胞因子驱动过程诱导。
朗格汉斯细胞瘤现有的治疗需要根据病情的不同选择不同的治疗方案,但由于不确定其发病原因,不能进行根本性的祛除病因,所以复发率较高。现有治疗方案有:手术、药物、化疗及放疗等。手术治疗:病人最初需要手术以进行活组织检查,之后需要进一步手术以移除所有朗格汉斯细胞。药物治疗:包括类固醇(泼尼松),非甾体类抗炎药(吲哚美辛),减少、消除局部炎症。化疗:通过干扰癌细胞生长或繁殖能力起作用的药物治疗,包括氮芥、长春碱、巯嘌呤等。放射疗法:使用来自专用机器的高能射线(辐射)来破坏或杀死异常细胞并缩小肿瘤,很少用于LCH治疗。
集落刺激因子1受体(CSF1R),也称为巨噬细胞集落刺激因子受体(M-CSFR),以及CD115,是一种在人类中由CSF1R基因编码的细胞表面蛋白。它是细胞因子CSF1的受体。它是单次通过I型膜蛋白,CSF1是控制巨噬细胞的产生,分化和功能的细胞因子。该受体介导该细胞因子的大部分生物效应。配体结合通过寡聚化和反式磷酸化过程激活CSF1R。CSF1R是酪氨酸激酶跨膜受体和酪氨酸蛋白激酶的CSF1/PDGF受体家族的成员。在阿尔茨海默病和脑损伤后的小胶质细胞中发现CSF1R1水平升高。受体表达增加导致小胶质细胞变得更活跃。CSF1R及其配体CSF1均在乳腺发育过程中起重要作用,可能参与乳腺癌的发生过程。CSF1R中的突变与慢性髓单核细胞白血病和M4型急性成髓细胞白血病相关。其酪氨酸激酶结构域的突变与球状遗传性弥漫性白质脑病相关。由于CSF1R在许多癌症和肿瘤相关巨噬细胞(TAM)中过表达,CSF1R抑制剂(和CSF1抑制剂)已被研究多年作为癌症或炎性疾病的可能治疗方法。截至2017年,临床试验中的CSF1R抑制剂包括:Pexidartinib,PLX7486,ARRY-382,JNJ-40346527,BLZ945,Emactuzumab,AMG820,IMC-CS4。(PD-0360324和MCS110是CSF1抑制剂)。另一种靶向、消耗TAM的CSF1R抑制剂是Cabiralizumab(cabira;FPA-008),它是一种单克隆抗体,目前正处于转移性胰腺癌的早期临床试验中。
发明内容
本发明的目的在于克服现有技术存在的不足,而提供一种以CSF1R作为药物靶点治疗肿瘤的药剂及其制备方法,涉及一个新的朗格汉斯细胞瘤或肿瘤相关巨噬细胞治疗位点,具体涉及以CSF1R作为靶点治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞,还涉及以CSF1R为靶点制备的拮抗剂:Ki 20227口服油剂及CSF1R抑制性单抗溶剂及其制备方法,包括使用它们制备药物作为靶向治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞的应用。
本发明的目的是通过如下技术方案来完成的。一种以CSF1R作为药物靶点治疗肿瘤的药剂,以CSF1R作为治疗肿瘤的药物靶点,药剂中含有CSF1R的拮抗剂。
所述的含有CSF1R的拮抗剂采用Ki 20227口服油剂,其中Ki 20227口服油剂中,Ki20227分子质量为480.54,其化学式为:C24H24N4O5S,Ki 20227口服油剂浓度为5mg/ml,结构式为:N-[4-[(6,7-Dimethoxy-4-quinolinyl)oxy]-2-methoxyphenyl]-N'-[1-(2-thiazolyl)ethyl]urea。
所述的含有CSF1R的拮抗剂是采用CSF1R抑制性单抗溶剂,含克隆号为AFS98的anti-mouseCSF1R的溶剂。
本发明同时公开了一种以CSF1R作为药物靶点治疗肿瘤的药剂的制备方法,步骤如下:
(1)、制备Ki 20227口服油剂的方法,将0.5gKi 20227溶于5ml50%的二甲基亚砜DMSO,溶解后加入到30ml的单油酸酯中,加热至40℃,充分混匀后加入65ml油酸,充分混匀,即制成5mg/ml的Ki 20227口服油剂;
(2)、制备CSF1R抑制性单抗溶剂的方法,将1mg克隆号为AFS98的anti-mouseCSF1R加入到2ml4℃含有0.1%BSA的PBS中,充分混匀后得到500μg/ml的CSF1R抑制性单抗溶剂,4℃保存。
本发明的有益效果为:与现有的技术相比,本发明应用CSF1R作为治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞或的靶点,可以使朗格汉斯细胞瘤患者或恶性肿瘤患者临床获益。本发明将CSF1R作为治疗朗格汉斯细胞瘤或抑制肿瘤相关巨噬细胞的靶点,应用其拮抗剂Ki20227口服油剂和CSF1R抑制性单抗溶剂,克服了对朗格汉斯细胞瘤或肿瘤相关巨噬细胞治疗的非选择性,达到靶向治疗朗格汉斯细胞瘤或靶向减少肿瘤相关巨噬细胞治疗恶性肿瘤、改善患者预后的目的,弥补了手术、激素药物、放化疗等治疗手段的不足,可以对微小病灶发挥作用;可以克服激素、放化疗等对机体的毒副作用。
目前的现有技术,拮抗CSF1R对朗格汉斯细胞瘤或抑制肿瘤相关巨噬细胞的功能是未知的,且以CSF1R为药物靶点治疗朗格汉斯细胞瘤或抑制肿瘤相关巨噬细胞的药剂及其制备也是未知的。
综上所述,本发明公开了两种以CSF1R为靶点的药剂及其制备方法。通过一系列的动物实验发现:CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂可以分别减少朗格汉斯细胞瘤中朗格汉斯细胞或分别减少肿瘤中肿瘤相关巨噬细胞,从而来抑制肿瘤的生长。这些实验数据均足以证明拮抗朗格汉斯细胞中的CSF1R可以显著的减少朗格汉斯细胞,具有成为肿瘤治疗有效靶点的潜能,同时这些实验数据均足以证明拮抗肿瘤相关巨噬细胞中的CSF1R可以显著的减少抗肿瘤相关巨噬细胞,具有成为肿瘤治疗有效靶点的潜能。CSF1R的拮抗剂在制备治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞的药物组合物上极具临床价值,制备的CSF1R拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂也已具备治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞的能力,其临床价值巨大,这都为朗格汉斯细胞瘤或肿瘤相关巨噬细胞的有效治疗提供了新的药物以及方法。
附图说明
图1是Ki 20227分子结构图。
图2是用分别用Ki 20227口服油剂和CSF1R抑制性单抗溶剂处理小鼠的模式图。
图3是应用流式细胞术分选朗格汉斯细胞的分选策略图,定义朗格汉斯细胞为CD45+CD11b+epCAM+。
图4是Ki20227口服油剂、anti-mouseCSF1R溶剂处理组和对照组的朗格汉斯细胞占比图。
A.Ki20227、anti-mouseCSF1R处理组和对照组的朗格汉斯细胞流式分选图,圈出的即为朗格汉斯细胞(CD45+CD11b+epCAM+)。
B.Ki20227、anti-mouseCSF1R处理组分别和对照组朗格汉斯细胞占比的t检验分析结果。(Ki20227,n=2;anti-mouseCSF1R,n=3;对照组,n=4;*P=0.0444,**P=0.0068)
图5是小鼠皮肤朗格汉斯细胞的免疫荧光实验图。
图6是小鼠肝癌模型制作流程图。
图7是小鼠结肠癌模型制作流程图。
图8是分别用Ki 20227口服油剂和CSF1R抑制性单抗溶剂处理肝癌和结肠癌模型小鼠的模式图。
图9是应用流式细胞术分选肿瘤相关巨噬细胞的分选策略图,定义抗肿瘤相关巨噬细胞为CD45+CD11b+CD14+。
图10是Ki20227口服油剂对照组分别对肝癌和结肠癌模型鼠处理后肿瘤相关巨噬细胞占比的流式图。
A.Ki20227口服油剂处理组和对照组的肿瘤相关巨噬细胞流式分选图,圈出的即为抗肿瘤相关巨噬细胞(CD45+CD11b+CD14+)。
B.Ki20227口服油剂处理组和对照组抗肿瘤相关巨噬细胞占比的t检验分析结果。(livertumor,n=2;liverantiCSF1R,n=3;****P<0.0001;colontumor,n=2;colonantiCSF1R,n=2;***P=0.0004)图11是CSF1R抑制性单抗溶剂对照组分别对肝癌和结肠癌模型鼠处理后肿瘤相关巨噬细胞占比的流式图。
A.CSF1R抑制性单抗溶剂处理组和对照组的肿瘤相关巨噬细胞流式分选图,圈出的即为抗肿瘤相关巨噬细胞(CD45+CD11b+CD14+)。
B.CSF1R抑制性单抗溶剂处理组和对照组抗肿瘤相关巨噬细胞占比的t检验分析结果。(livertumor,n=2;liverantiCSF1R,n=2;**P=0.0066;colontumor,n=2;colonantiCSF1R,n=2;*P=0.0146)
具体实施方式
为了便于理解,一下将通过具体的附图和实例进一步阐述本发明内容。需要特别指出的是,具体实例和附图仅是为了说明,本领域的普通技术人员可以根据本文说明,在本发明的范围内对本发明做出各种各样的修改和改变,这些修改和改变也纳入本发明的范围。
本发明解决的技术问题是提供一种以CSF1R作为药物靶点在朗格汉斯细胞瘤或肿瘤相关巨噬细胞治疗中的应用及提供两种采用CSF1R为靶点治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞的药物制剂及其制备方法。
为了解决上述技术问题,本发明提供一种CSF1R的用途:作为朗格汉斯细胞瘤或肿瘤相关巨噬细胞的治疗靶点,来制备药剂及制备方法。
本发明提供了CSF1R作为药物靶点在朗格汉斯细胞瘤或肿瘤相关巨噬细胞的治疗药物中的应用,所述CSF1R是酪氨酸激酶跨膜受体和酪氨酸蛋白激酶的CSF1/PDGF受体家族的成员。
CSF1R在褐家鼠(Rattus norvegicus)中具体的蛋白质序列为:
CSF1R在人中具体的蛋白序列为:
本发明提供了一种药剂以CSF1R作为药物靶点治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞,即以Ki20227口服油剂来拮抗CSF1R。Ki 20227作为CSF1R的拮抗剂。
Ki 20227分子质量为480.54,其化学式为:C24H24N4O5S,结构式为N-[4-[(6,7-Dimethoxy-4-quinolinyl)oxy]-2-methoxyphenyl]-N'-[1-(2-thiazolyl)ethyl]urea(见图1)。Ki 20227口服油剂浓度为5mg/ml,其溶剂含有:二甲基亚砜(DMSO)、单油酸酯及油酸。
为了应用Ki 20227,本发明提供了一种Ki 20227口服油剂的制备方法:将0.5gKi20227溶于5ml50%的DMSO,溶解后加入到30ml的单油酸酯中,加热至40℃,充分混匀后加入65ml油酸,充分混匀,即制成5mg/ml的Ki 20227口服油剂。
本发明提供了另一种药剂以CSF1R作为药物靶点治疗朗格汉斯细胞瘤或肿瘤相关巨噬细胞,即以CSF1R抑制性单抗溶剂来拮抗CSF1R。CSF1R抑制性单抗作为CSF1R的拮抗剂。
CSF1R抑制性单抗为克隆号为AFS98的anti-mouseCSF1R(CD115)。CSF1R抑制性单抗溶剂浓度为500μg/ml,其溶剂含有:0.1%牛血清蛋白(BSA)、磷酸盐缓冲液(PBS)。
为了应用CSF1R抑制性单抗,本发明提供了一种CSF1R抑制性单抗溶剂的制备方法:将1mg克隆号为AFS98的anti-mouseCSF1R(CD115)加入到2ml4℃含有0.1%BSA的PBS中,充分混匀后得到500μg/ml的CSF1R抑制性单抗溶剂,4℃保存。
本发明提供的方案具体如下:
实例1:本发明提供了一个新的朗格汉斯细胞瘤治疗的靶点——CSF1R。具体而言,用CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂分别处理小鼠(如图2),运用流式细胞术筛选出朗格汉斯细胞(CD45+CD11b+epCAM+)(如图3),得到朗格汉斯细胞占比和对照组进行比较显示,Ki 20227和CSF1R抑制性单抗处理的两组小鼠的朗格汉斯细胞占比均显著性减少(如图4A、4B)。
进一步,利用免疫荧光技术验证了在Ki 20227和CSF1R抑制性单抗处理后小鼠皮肤中朗格汉斯细胞较处理前均明显减少(如图5)。
本发明提供的靶向抑制朗格汉斯细胞瘤的两种药剂,该药剂靶向拮抗CSF1R,从而抑制朗格汉斯细胞。制备Ki 20227口服油剂和CSF1R抑制性单抗溶剂的方法。
1.制备Ki 20227口服油剂的方法。
实施方法:将0.5gKi 20227溶于5ml50%的DMSO,溶解后加入到30ml的单油酸酯中,加热至40℃,充分混匀后加入65ml油酸,充分混匀,即制成5mg/ml的Ki 20227口服油剂。
2.制备CSF1R抑制性单抗溶剂的方法。
实施方法:将1mg克隆号为AFS98的anti-mouseCSF1R(CD115)加入到2ml4℃含有0.1%BSA的PBS中,充分混匀后得到500μg/ml的CSF1R抑制性单抗溶剂,4℃保存。
CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂可以减少朗格汉斯细胞瘤中朗格汉斯细胞。
1.Ki 20227口服油剂和CSF1R抑制性单抗溶剂对小鼠朗格汉斯细胞影响的流式细胞术分析。
实施方法:分别用CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂处理小鼠,Ki 20227口服溶剂的处理方法为1ml的5mg/mlKi 20227口服油剂每三天灌胃一次,持续一个月;CSF1R抑制性单抗处理方法为200μl的500mg/lCSF1R抑制性单抗溶剂每三天腹腔内注射一次,持续两星期。然后分离小鼠皮肤组织细胞,利用流式细胞术计算出朗格汉斯细胞细胞(CD45+CD11b+epCAM+)占CD45+细胞的百分比。
结果分析:如图4所示,相比对照组,Ki 20227处理组的朗格汉斯细胞占CD45+细胞的百分比显著性降低;相比对照组,CSF1R抑制性单抗处理组的朗格汉斯细胞占CD45+细胞的百分比也显著性降低。以上实验结果说明,CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂能明显抑制甚至杀死小鼠皮肤中的朗格汉斯细胞。
2.Ki 20227口服油剂和CSF1R抑制性单抗溶剂对小鼠处理前后,小鼠皮肤朗格汉斯细胞的免疫荧光分析。
实施方法:应用免疫荧光技术分别对Ki 20227口服油剂和CSF1R抑制性单抗溶剂处理组小鼠的皮肤在处理前和处理后进行处理,在荧光显微镜下观察荧光强度以反映朗格汉斯细胞的数量。
结果分析:相比处理前,Ki 20227处理组的朗格汉斯细胞显著性减少;相比处理前,CSF1R抑制性单抗处理组的朗格汉斯细胞显著性减少。以上实验结果说明,CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂能明显抑制甚至杀死小鼠皮肤中的朗格汉斯细胞。
实例2:本发明提供了两种以CSF1R为治疗靶点抑制肿瘤相关巨噬细胞的药剂及其制备方法——Ki20227口服油剂和CSF1R抑制性单抗溶剂。具体而言,用CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂分别同时处理肝癌和结肠癌小鼠模型(如图8),运用流式细胞术筛选出肿瘤相关巨噬细胞(CD45+CD11b+CD14+)(如图9-11),得到肿瘤相关巨噬细胞占比和对照组进行比较显示,在肝癌和大肠癌小鼠模型中分别用Ki 20227和CSF1R抑制性单抗处理的小鼠的肿瘤相关巨噬细胞占比均显著性减少(如图10A、10B、11A、11B)。
本发明提供的两种药剂靶向抑制肿瘤相关巨噬细胞的方法,该方法靶向拮抗CSF1R,从而抑制肿瘤相关巨噬细胞。
制备Ki 20227口服油剂和CSF1R抑制性单抗溶剂的方法。
1.制备Ki 20227口服油剂的方法。
实施方法:将0.5gKi 20227溶于5ml50%的DMSO,溶解后加入到30ml的单油酸酯中,加热至40℃,充分混匀后加入65ml油酸,充分混匀,即制成5mg/ml的Ki 20227口服油剂。
2.制备CSF1R抑制性单抗溶剂的方法。
实施方法:将1mg克隆号为AFS98的anti-mouseCSF1R(CD115)加入到2ml4℃含有0.1%BSA的PBS中,充分混匀后得到500μg/ml的CSF1R抑制性单抗溶剂,4℃保存。
制备肝癌和结肠癌小鼠模型的方法。
3.制作肝癌小鼠模型的方法
实施方法:鼠源为C57BL/6J 7-9周雄性老鼠,将PT3-EF1a-C-Myc,PT/Caggs-NRas-V12两种质粒同时稀释于磷酸缓冲液中(phosphate-buffered saline,PBS),按0.1mg/g体重在5-7秒内快速从尾静脉注入。喂养3星期。
4.制作结肠癌小鼠模型的方法
实施方法:鼠源为转基因APCMin/+老鼠,用含有1.5%DSS(dextran sodium sulfate)水喂养一周,再用普通水喂养3周。
CSF1R的拮抗剂Ki 20227口服油剂和CSF1R抑制性单抗溶剂可以减少肝癌和结肠癌中肿瘤相关巨噬细胞。
1.Ki 20227口服油剂对小鼠肿瘤相关巨噬细胞影响的流式细胞术分析。
实施方法:用CSF1R的拮抗剂Ki 20227口服油剂处理小鼠,Ki 20227口服溶剂的处理方法为1ml的5mg/mlKi 20227口服油剂每三天灌胃一次,持续一个月。然后分离小鼠皮肤组织细胞,利用流式细胞术计算出肿瘤相关巨噬细胞细胞(CD45+CD11b+CD14+)占比。
结果分析:如图10所示,相比肝癌小鼠对照组,Ki 20227处理组的肝癌中肿瘤相关巨噬细胞占比显著性降低;相比结肠癌对照组,Ki 20227处理组的结肠癌中肿瘤相关巨噬细胞占比显著性降低。以上实验结果说明,CSF1R的拮抗剂Ki 20227口服油剂能明显抑制甚至杀死小鼠皮肤中的肿瘤相关巨噬细胞。
2.CSF1R抑制性单抗溶剂对小鼠肿瘤相关巨噬细胞影响的流式细胞术分析。
实施方法:用CSF1R抑制性单抗溶剂处理小鼠,CSF1R抑制性单抗处理方法为200μl的500mg/lCSF1R抑制性单抗溶剂每三天腹腔内注射一次,持续两星期。然后分离小鼠皮肤组织细胞,利用流式细胞术计算出肿瘤相关巨噬细胞细胞(CD45+CD11b+CD14+)占比。
结果分析:如图10所示,相比肝癌小鼠对照组,CSF1R抑制性单抗溶剂处理组的肝癌中肿瘤相关巨噬细胞占比显著性降低;相比结肠癌对照组,CSF1R抑制性单抗溶剂处理组的结肠癌中肿瘤相关巨噬细胞占比显著性降低。以上实验结果说明,CSF1R抑制性单抗溶剂处理组能明显抑制甚至杀死小鼠皮肤中的肿瘤相关巨噬细胞。
可以理解的是,对本领域技术人员来说,对本发明的技术方案及发明构思加以等同替换或改变都应属于本发明所附的权利要求的保护范围。
Claims (4)
1.一种以CSF1R作为药物靶点治疗肿瘤的药剂,其特征在于:以CSF1R作为治疗肿瘤的药物靶点,药剂中含有CSF1R的拮抗剂。
2.根据权利要求1所述的以CSF1R作为药物靶点治疗肿瘤的药剂,其特征在于:所述的含有CSF1R的拮抗剂采用Ki 20227口服油剂,其中Ki 20227口服油剂中,Ki 20227分子质量为480.54,其化学式为:C24H24N4O5S,Ki 20227口服油剂浓度为5mg/ml,结构式为:
N-[4-[(6,7-Dimethoxy-4-quinolinyl)oxy]-2-methoxyphenyl]-N'-[1-(2-thiazolyl)ethyl]urea。
3.根据权利要求1所述的以CSF1R作为药物靶点治疗肿瘤的药剂,其特征在于:所述的含有CSF1R的拮抗剂是采用CSF1R抑制性单抗溶剂,含克隆号为AFS98的anti-mouseCSF1R的溶剂。
4.一种制备如权利要求1所述的以CSF1R作为药物靶点治疗肿瘤的药剂的方法,其特征在于:该方法主要包括:
(1)、制备Ki 20227口服油剂的方法,将0.5gKi 20227溶于5ml50%的二甲基亚砜DMSO,溶解后加入到30ml的单油酸酯中,加热至40℃,充分混匀后加入65ml油酸,充分混匀,即制成5mg/ml的Ki 20227口服油剂;
(2)、制备CSF1R抑制性单抗溶剂的方法,将1mg克隆号为AFS98的anti-mouseCSF1R加入到2ml4℃含有0.1%BSA的PBS中,充分混匀后得到500μg/ml的CSF1R抑制性单抗溶剂,4℃保存。
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