CN115068613A - 泛tki在治疗hr阳性her2低表达乳腺癌的应用 - Google Patents
泛tki在治疗hr阳性her2低表达乳腺癌的应用 Download PDFInfo
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Abstract
本发明提供了泛HER受体酪氨酸激酶抑制剂在制备治疗HR+HER2low乳腺癌的药物中的应用。本发明发现泛HER受体酪氨酸激酶抑制剂可以明显增加HR+HER2low乳腺癌细胞对ET加CDK4/6i治疗的敏感性，通过阻滞G1期抑HR+HER2low乳腺癌细胞的增殖，且明显抑制HR+HER2low人乳腺癌异种移植小鼠肿瘤的生长和转移。说明泛HER受体酪氨酸激酶抑制剂可使HR+HER2low乳腺癌患者在ET加CDK4/6i疗法中受益，可用于HR+HER2low乳腺癌患者的治疗。
Description
技术领域
本发明属于生物医药技术领域，具体涉及泛TKI在治疗HR阳性HER2低表达乳腺癌的应用。
背景技术
我国使用免疫组化(IHC)和荧光原位杂交技术(FISH)将乳腺癌中HER2的表达情况分为HER2-和HER2+，其中HR+HER2-乳腺癌是最常见的乳腺癌亚型。根据2018年ASCO/CAP指南，根据染色强度和完整细胞膜染色数的百分比，将HER2检测的IHC结果分为IHC 0、IHC 1+、IHC 2+和IHC 3+；其中IHC 0代表无染色或微弱染色的不完整细胞膜染色且染色肿瘤细胞小于等于10％；IHC 1+代表微弱染色的不完整细胞膜染色且染色肿瘤细胞大于10％；IHC2+代表微弱至中度完整细胞膜染色，且染色肿瘤细胞大于10％；IHC 3+代表高度完整细胞膜染色，且染色肿瘤细胞大于10％。HER2+包括IHC 2+且FISH+及IHC 3+，HER2-包括IHC 0或IHC 1+或IHC 2+且FISH阴性。而根据IHC和FISH检测结果可将HER2-进一步划分为HER2low和HER2-0。HER2low定义为：IHC 1+或IHC 2+且FISH阴性；HER2-0定义为：IHC 0。临床数据表明，超过50％-70％的HR+HER2-患者呈现HER2low状态。与HR+HER2-0乳腺癌患者相比，HR+HER2low乳腺癌患者表现出更大的肿瘤体积、更高的组织病理学分级和更高的Ki67表达，且更长出现腋窝***转移。
内分泌治疗(ET)+CDK4/6抑制剂(CDK4/6i)是目前HR+HER2-乳腺癌的一、二线标准治疗方案。但实际临床治疗中发现，部分HR+HER2-乳腺癌患者会出现ET耐药和CDK4/6i敏感性降低，目前并不清楚具体的原因和解决的方案。
发明内容
基于此，本发明的目的在于提供泛HER受体酪氨酸激酶抑制剂在治疗HR阳性HER2低表达(HR+HER2low)乳腺癌的应用。
为达到上述目的，本发明采用如下技术方案。
泛HER受体酪氨酸激酶抑制剂(泛TKI)在制备治疗HR+HER2low乳腺癌的药物中的应用，所述HER2low判断标准为ⅰ或ⅱ：ⅰ：IHC 1+；ⅱ：IHC 2+且FISH阴性。
在一些实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种。
在一些优选的实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼。
本发明还提供了药物组合物在制备治疗HR+HER2low乳腺癌的药物中的应用，所述药物组合物包含泛HER受体酪氨酸激酶抑制剂、CDK4/6抑制剂和***受体拮抗剂；所述HER2low判断标准为ⅰ或ⅱ：ⅰ：IHC 1+；ⅱ：IHC 2+且FISH阴性。
在一些实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种。
在一些优选的实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼。
在一些实施例中，所述***受体拮抗剂选自氟维司群。
在一些实施例中，所述CDK4/6抑制剂选自帕博西尼。
本发明还提供了一种治疗HR+HER2low乳腺癌的药物，包含泛HER受体酪氨酸激酶抑制剂、CDK4/6抑制剂、***受体拮抗剂和药物学上可接受的载体。
在一些实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种；和/或，所述CDK4/6抑制剂选自帕博西尼；和/或，所述***受体拮抗剂选自氟维司群。
在一些优选的实施例中，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼。
本发明发现对ET耐药和CDK4/6i敏感性降低的HR+HER2-乳腺癌患者主要为HR+HER2low乳腺癌患者，并进一步经过研究发现泛HER受体酪氨酸激酶抑制剂可以明显增加HR+HER2low乳腺癌细胞对ET加CDK4/6i治疗的敏感性，通过阻滞G1期抑HR+HER2low乳腺癌细胞的增殖，且明显抑制HR+HER2low人乳腺癌异种移植小鼠肿瘤的生长和转移。说明泛HER受体酪氨酸激酶抑制剂可使HR+HER2low乳腺癌患者在ET加CDK4/6i疗法中受益，可用于HR+HER2low乳腺癌患者的治疗。
附图说明
图1为ZR-75-1、T47D、BT474细胞中HER2的表达结果图。
图2为ET、CDK4/6i、奈拉替尼对ZR-75-1细胞增殖影响结果图。
图3为奈拉替尼加ET加CDK4/6i对ZR-75-1细胞增殖影响结果图。
图4为奈拉替尼加ET加CDK4/6i对ZR-75-1细胞周期影响结果图。
图5为奈拉替尼加ET加CDK4/6i抑制人乳腺癌异种移植小鼠肿瘤生长。
具体实施方式
本发明下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。实施例中所用到的各种常用化学试剂，均为市售产品。
除非另有定义，本发明所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本发明的说明书中所使用的术语只是为了描述具体的实施例的目的，不用于限制本发明。
本发明的术语“包括”和“具有”以及它们任何变形，意图在于覆盖不排他的包含。例如包含了一系列步骤的过程、方法、装置、产品或设备没有限定于已列出的步骤或模块，而是可选地还包括没有列出的步骤，或可选地还包括对于这些过程、方法、产品或设备固有的其它步骤。
在本发明中提及的“多个”是指两个或两个以上。“和/或”，描述关联对象的关联关系，表示可以存在三种关系，例如，A和/或B，可以表示：单独存在A，同时存在A和B，单独存在B这三种情况。字符“/”一般表示前后关联对象是一种“或”的关系。
下面结合具体实施例进行说明。以下实施例中使用的泛HER受体酪氨酸激酶抑制剂为奈拉替尼(Neratinib)，ET为氟维司群(Fulvestrant)，CDK4/6i为帕博西尼(Palbociclib)。
实施例1
本实施例研究泛HER受体酪氨酸激酶抑制剂奈拉替尼联合ET加CDK4/6i对HR+HER2low乳腺癌的疗效。
一、实验方法
1ZR-75-1细胞中HER2的表达
设阳性对照组HR+HER2+(BT474细胞)、阴性对照组HR+HER2-0(T47D细胞)和实验组HR+HER2low(ZR-75-1细胞)，使用细胞免疫荧光检测3种细胞HER2的表达情况。具体操作：在96孔板每孔种5×103个细胞于细胞培养箱中培养；24h后取出PBS洗两次，加4％PFA多聚甲醛室温固定20min；PBS洗两次，加0.5％TX-100室温通透细胞膜20min；PBS洗两次，加5％山羊血清封闭1h；甩干后加稀释在5％BSA中的一抗HER2(稀释比1：200)4℃过夜；复温45min，PBS洗两次，加稀释在PBS中的荧光二抗兔抗(稀释比1：1000)避光37℃30min；PBS洗两次，加DAPI染细胞核5min；抽掉上清，加抗荧光淬灭剂在荧光显微镜下观察。
2奈拉替尼对ZR-75-1细胞增殖实验
分别检测奈拉替尼、ET和CDK4/6i对ZR-75-1细胞增殖的影响：设奈拉替尼浓度0、0.1nM、1nM、10nM、100nM、1000nM、10000nM七组，设ET 0、0.1nM、1nM、10nM、100nM、1000nM、10000nM、100000nM八组，设CDK4/6i 0、25nM、50nM、100nM、200nM、400nM、800nM、1600nM、3200nM、10000nM十组。使用CCK8实验检测不同时间不同浓度的奈拉替尼对ZR-75-1细胞增殖的影响。具体操作：在96孔板每孔种1×103个细胞于细胞培养箱中培养；24h后按照以上分组进行药物干预ZR-75-1细胞，分别24h、48h、72h于多功能酶标仪波长450nm处检测OD值。
3奈拉替尼联合ET加CDK4/6i后对ZR-75-1细胞增殖实验
ET(Fulvestrant)标准浓度100nM下，设浓度梯度比例的CDK4/6i和奈拉替尼，使用CCK8实验检测24h、48h、72h使用ET、ET加CDK4/6i以及奈拉替尼联合ET加CDK4/6i对ZR-75-1细胞增殖的影响。实验结果发现48h的结果能够显著体现联合用药方案与其他方案的差异。随后使用同样的分组，在48h检测各组用药对不同亚型细胞T47D和BT474细胞增殖的影响。具体操作与步骤2相同。
4奈拉替尼联合ET加CDK4/6i后对ZR-75-1细胞周期实验
根据以上实验计算联合指数，寻找最合适的联合用药浓度，确定联合ET(100nM)+CDK4/6i(125nM)后的奈拉替尼最佳浓度是125nM，设Control组(药物溶剂DMSO)、ET(100nM)+CDK4/6i(100nM)组、ET(100nM)+CDK4/6i(100nM)+奈拉替尼(125nM)组共三组，使用流式细胞术检测不同组对ZR-75-1细胞周期的影响。具体操作：在6孔板每孔种5×105个细胞于细胞培养箱中培养；24h后按照以上分组进行药物干预ZR-75-1细胞；24h后取出PBS洗一次，加不含EDTA胰酶消化细胞，离心；加70％冷乙醇(PBS稀释)4℃固定过夜；PBS洗两次，加细胞周期染色试剂避光室温孵育40min，以标准程序用流式细胞仪检测，结果用细胞周期拟和软件ModFit分析。
5奈拉替尼联合ET加CDK4/6i后对乳腺癌小鼠模型实验
进行小鼠去卵巢手术后，建立HR+HER2low人乳腺癌细胞异种移植小鼠模型，随机分组：Control组、ET加CDK4/6i组、ET加CDK4/6i+奈拉替尼组，每组8只。参考药物临床前研究，小鼠的给药剂量分别为：ET(Fulvestrant，100mg/kg，肌注，14天一次)；CDK4/6i(Palbociclib，150mg/kg，灌胃，1天一次)；奈拉替尼(Neratinib，30mg/kg，灌胃，1天一次)；Control组使用药物溶剂生理盐水，给药方式如前所述。研究ET加CDK4/6i及ET加CDK4/6i+奈拉替尼对小鼠状态(体重、存活率)肿瘤状况(瘤体数目、体积、重量)、转移情况(转移瘤数目、转移部位)的改变。当肿瘤体积达到1500mm3时，按照动物伦理指南处死小鼠并收集小鼠肿瘤，免疫组化及western blot检测ET加CDK4/6i及ET加CDK4/6i+奈拉替尼组小鼠肿瘤组织中Ki67、HER2、CDK4的表达水平变化。
二、实验结果
使用细胞免疫荧光检测了三组乳腺癌细胞系(ZR-75-1、T47D、BT474)中HER2的表达，发现BT474细胞系表现出高HER2表达，三组乳腺癌细胞荧光强度值从强到弱为T47D细胞<ZR-75-1细胞<BT474细胞(图1)。
奈拉替尼、ET和CDK4/6i对ZR-75-1细胞增殖的影响结果如图2所示。奈拉替尼加ET加CDK4/6i组有效抑制HR+HER2low乳腺癌细胞的增殖，且最佳联合浓度为：ET(Fulvestrant:100nM)+CDK4/6i(Palbociclib:100nM)+TKI(Neratinib:125nM)。检测结果表明在ET加CDK4/6i联合奈拉替尼后能明显抑制ZR-75-1的增殖，但对T47D的ET加CDK4/6i增效作用不明显(图3)。
流式细胞周期结果显示奈拉替尼(125nM)+ET(100nM)+CDK4/6i(100nM)组是通过阻滞G1期抑制ZR-75-1细胞增殖，从而抑制乳腺癌的生长(图4)。
HR+HER2low人乳腺癌细胞异种移植小鼠模型结果(图5)显示，与Control组相比，奈拉替尼加ET加CDK4/6i组小鼠肿瘤体积明显小于Control组和ET加CDK4/6i组，脾、肝等器官未发现肿瘤转移。表明奈拉替尼加ET加CDK4/6i可明显抑制人乳腺癌异种移植小鼠肿瘤生长，且对人乳腺癌异种移植小鼠肿瘤的抑制作用优于ET加CDK4/6i组。
上述结果表明，奈拉替尼可以有效提升ET加CDK4/6i疗法对HR+HER2low乳腺癌的治疗效果。
以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对以上实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。
Claims (10)
1.泛HER受体酪氨酸激酶抑制剂在制备治疗HR+HER2low乳腺癌的药物中的应用，其特征在于，所述HER2low判断标准为ⅰ或ⅱ：ⅰ：IHC 1+；ⅱ：IHC 2+且FISH阴性。
2.如权利要求1所述的应用，其特征在于，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种。
3.如权利要求1所述的应用，其特征在于，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼。
4.药物组合物在制备治疗HR+HER2low乳腺癌的药物中的应用，其特征在于，所述药物组合物包含泛HER受体酪氨酸激酶抑制剂、CDK4/6抑制剂和***受体拮抗剂；所述HER2low判断标准为ⅰ或ⅱ：ⅰ：IHC 1+；ⅱ：IHC 2+且FISH阴性。
5.如权利要求4所述的应用，其特征在于，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种。
6.如权利要求4所述的应用，其特征在于，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼。
7.如权利要求4所述的应用，其特征在于，所述***受体拮抗剂选自氟维司群。
8.如权利要求4所述的应用，其特征在于，所述CDK4/6抑制剂选自帕博西尼。
9.一种治疗HR+HER2low乳腺癌的药物，其特征在于，包含泛HER受体酪氨酸激酶抑制剂、CDK4/6抑制剂、***受体拮抗剂和药物学上可接受的载体。
10.如权利要求9所述的药物，其特征在于，所述泛HER受体酪氨酸激酶抑制剂选自奈拉替尼和吡咯替尼中的至少一种；和/或，所述CDK4/6抑制剂选自帕博西尼；和/或，所述***受体拮抗剂选自氟维司群。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170363630A1 (en) * | 2014-12-08 | 2017-12-21 | George Mason University | Methods for breast cancer treatment |
| CN109310754A (zh) * | 2016-03-15 | 2019-02-05 | 梅里麦克制药股份有限公司 | 使用包含抗erbb3抗体的联合疗法治疗er+、her2-、hrg+乳腺癌的方法 |
| CN111184863A (zh) * | 2018-11-15 | 2020-05-22 | 江苏恒瑞医药股份有限公司 | 酪氨酸激酶抑制剂、cdk4/6抑制剂、serd联合在制备***的药物中的用途 |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170363630A1 (en) * | 2014-12-08 | 2017-12-21 | George Mason University | Methods for breast cancer treatment |
| CN109310754A (zh) * | 2016-03-15 | 2019-02-05 | 梅里麦克制药股份有限公司 | 使用包含抗erbb3抗体的联合疗法治疗er+、her2-、hrg+乳腺癌的方法 |
| CN111184863A (zh) * | 2018-11-15 | 2020-05-22 | 江苏恒瑞医药股份有限公司 | 酪氨酸激酶抑制剂、cdk4/6抑制剂、serd联合在制备***的药物中的用途 |
Non-Patent Citations (3)
| Title |
|---|
| C DENKERT等: "Clinical and molecular characteristics of HER2-low-positive breast cancer: pooled analysis of individual patient data from four prospective, neoadjuvant clinical trials", 《LANCET ONCOL》, vol. 2021, no. 22, pages 1151 - 1161 * |
| ONSUM 等: "Single-Cell Quantitative HER2 Measurement Identifies Heterogeneity and Distinct Subgroups within Traditionally Defined HER2-Positive Patients", 《AM J PATHOL》, vol. 183, no. 5, pages 1446 - 1460, XP055238425, DOI: 10.1016/j.ajpath.2013.07.015 * |
| 萨拉•赫维茨等, 辽宁科学技术出版社 * |
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