CN115850264A - 新型protac化合物及其在抗癌药物中的应用 - Google Patents
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Abstract
本发明公开了新型PROTAC化合物及其在抗癌药物中的应用。本发明中构建的蛋白降解靶向嵌合体(PROTAC)可使靶蛋白BCR‑ABL泛素化,通过泛素‑蛋白酶体降解靶蛋白,避免由于靶点突变产生的耐药性。本发明为构建蛋白降解靶向嵌合体提供了新选择,构建的蛋白降解靶向嵌合体在制备预防和/或治疗癌症药物中有良好的应用。
Description
技术领域
本发明属于生物医药领域,具体涉及以达沙布韦(Dasabuvir)衍生物/结构类似物作为E3连接酶新型配体构建的系列新型蛋白降解靶向嵌合体(PROTAC)在制备预防和/或治疗癌症药物中的用途。
背景技术
蛋白降解靶向嵌合体(proteolysis targeting chimeric molecule, PROTAC)作为一类新的药物可降解多种与人类疾病相关的蛋白质,具有高效能,高选择性,以及靶向“不可成药”蛋白等优势。PROTAC技术可用于靶向多种癌症靶点,包括对抗实体瘤和恶性血癌的不同靶点,且以靶向依赖的方式显示出对某些肿瘤细胞的高效杀伤力,例如靶向BRD4、BTK、BCR-ABL和CDK-6等靶点的PROTACs已经显示了治疗白血病的潜能;而靶向AR、ER、FAK、P38的ROTACs正在被开发用于治疗多种不同的实体瘤;靶向BCL-XL和ALK 的PROTACs还显示出了广谱的抗肿瘤活性,在体外和异种移植模型中均能有效杀伤白血病和实体瘤细胞。
科学家在人类基因组中发现了600多个E3连接酶,包括CRBN、VHL、MDM2、IAPs、DCAF15、DCAF16、RNF4和RNF114 等。文献报道的应用到PROTAC中的E3连接酶主要有CRBN、VHL、cIAP和MDM2,而效果较好、使用频次最高的E3连接酶主要是CRBN和VHL两种。2010年,Handa及其同事的研究发现E3连接酶Cereblon(CRBN)为沙利度胺(thalidomide)的主要蛋白质靶点;Cereblon是由人类CRBN基因编码的蛋白,CRBN同源基因是高度保守的,这表明它在生理学中的重要性。
目前,常用的CRBN E3连接酶的配体为结构高度相似的沙利度胺(thalidomide)、来那度胺(lenalidomide)和泊马度胺(pomalidomide),而沙利度胺是一种致畸药物“反应停”,妊娠女性服用后可能导致胎儿出生缺陷,甚至死亡,具有较大的毒副作用。达沙布韦(Dasabuvir)(在我国上市的商品名为“易奇瑞”)属于非核苷类聚合酶抑制剂,具有抑制丙肝病毒NS5B聚合酶的作用,主要用来抑制丙肝病毒RNA的复制,本发明以达沙布韦(Dasabuvir)衍生物/结构类似物作为E3连接酶新型配体构建的系列新型蛋白降解靶向嵌合体(PROTAC),可以靶向降解BCR-ABL等癌蛋白,一定程度上起到抗癌(包括但不局限性慢性髓系白血病等)的效果。
发明内容
本发明的目的在于提供一类小分子化合物达沙布韦(Dasabuvir)的衍生物/结构类似物作为E3连接酶CRBN新型配体构建蛋白降解靶向嵌合体(PROTAC)在制备预防和/或治疗癌症药物中的用途。
本发明的目的通过如下技术方案实现:
筛选获得达沙布韦作为E3连接酶新型配体,所述达沙布韦分子如式Ⅲ,基于达沙布韦,我们优化合成了可适合于构建PROTAC的新型E3连接酶新型配体结构单元,分子式如Ⅳ,Ⅴ:
基于结构式Ⅳ,Ⅴ,合成了一种新型PROTAC化合物,靶向降解BCR-ABL蛋白,其分子结构分别如式Ⅰ,Ⅱ:
式Ⅰ
式Ⅱ
所述构建PROTAC可降解蛋白包括但不局限于癌症相关突变癌蛋白如BCR-ABL等。BCR-ABL是一种融合癌蛋白, 具有高度酪氨酸激酶活性,细胞过度增殖而使细胞调控发生紊乱,进而致癌。
本发明还公开了一类抗癌药物组合物,含有所述构建蛋白降解靶向嵌合体和药学上可接受的辅料。
本发明具有以下优点:
1、 本发明提供了一类与市场及科研领域常用的CRBN的配体沙利度胺(thalidomide)、来那度胺(lenalidomide)、泊马度胺(pomalidomide)等化学结构不同的E3连接酶新型配体达沙布韦(Dasabuvir)而衍生出的适合于构建PROTAC的新型CRBN配体。
2、本发明提供了一类基于达沙布韦(Dasabuvir)的衍生物/结构类似物构建双功能小分子(如使BCR-ABL蛋白靶向降解的化合物Bcr-abl-PROTAC-014, Bcr-abl-PROTAC-020),分子一端结合靶蛋白BCR-ABL,另一端通过达沙布韦(Dasabuvir)衍生物/结构类似物结合 E3连接酶CRBN,将靶蛋白和E3连接酶拉近使靶蛋白泛素化,通过泛素-蛋白酶体途径降解靶蛋白,可避免由于靶点突变产生的耐药性。
3、本发明提供了一类新的PROTAC设计及合成路线为构建蛋白降解靶向嵌合体(PROTAC)提供了新选择。
附图说明
图1为 Bcr-abl-PROTAC-014用药24小时处理K562癌细胞后(40 μM)靶蛋白降解图;
图2为 Bcr-abl-PROTAC-020用药24小时处理K562癌细胞后(40 μM)靶蛋白降解图;
图3为Bcr-abl-PROTAC-014用药处理48小时后K562癌细胞(10 μM)细胞活力图;
图4为Bcr-abl-PROTAC-020用药处理48小时后K562癌细胞(10 μM)细胞活力图。
实施方式
本发明的目的在于提供一种小分子化合物达沙布韦(Dasabuvir)的衍生物/结构类似物作为E3连接酶CRBN新型配体构建新型蛋白降解靶向嵌合体(PROTAC)在制备预防和/或治疗癌症药物中的用途。
本发明的目的通过如下技术方案实现,下述实施例将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅是本发明的部分实施例,而不是全部。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
Bcr-abl-PROTAC-014的制备
N-(2-氯-6-甲基苯基)-2-((6-(4-(5-(5-(2,4-二氧代-3,4-二氢嘧啶-1(2H)-基)吡啶-2-基)氨基)-5-氧代戊酰基)哌嗪-1-基)-2-甲基嘧啶-4-基)氨基氨基)噻唑-5-甲酰胺合成路线:
步骤一:称取原料2-氨基-5-碘吡啶(2.0 g, 9.1 mmol)于100 mL单颈瓶中,室温下加入环戊二酸酐(1.14 g, 10.0 mmol),无水四氢呋喃(50 mL),升温至回流反应12 h。TLC监测反应完全后,冷却至室温,抽滤,收集滤饼,干燥后得到目标产物中间体1-1。(2.7g, 88.6%)HRMS (ESI): m/z [M+H]+.C10H12IN2O2计算值334.9887,实测值334.9896。
步骤二:称取中间体1-1(1.0 g, 3.0 mmol)于100 mL单颈瓶中,室温下加入达沙替尼中间体原料(1.45 g, 3.3 mmol),HATU(1.25 g, 3.3 mmol),DIPEA(0.85 g, 6.6mmol),无水DMF(20 mL),室温搅拌过夜,TLC监测反应完全后,加水并使用二氯甲烷萃取2-3次,合并有机层,水洗,饱和食盐水洗,真空浓缩后将残余物拌入硅胶制砂,柱层析(DCM:MeOH = 30:1)得到中间体1-2纯品。(0.7 g, 30.7%)HRMS (ESI): m/z [M+H]+.C30H32ClIN9O3S计算值760.1077,实测值760.1081。
步骤三:称取中间体1-2(0.6 g, 0.79 mmol)于50 mL单颈瓶中,室温下加入尿嘧啶(0.13 g, 1.19 mmol),碘化亚铜(0.23 g, 1.19 mmol),L-脯氨酸(0.14 g, 1.19mmol),磷酸三钾(0.25 g, 1.19 mmol),无水DMSO(10 mL),N2气保护下60 oC搅拌反应过夜,TLC监测反应完全后,加水并使用二氯甲烷萃取2-3次,合并有机层,水洗,饱和食盐水洗,真空浓缩后将残余物拌入硅胶制砂,柱层析(DCM:MeOH = 20:1)得到目标产物bcr-abl-PROTAC-014纯品。(0.1 g, 17.0%)1HNMR(400 MHz, CDCl3): δ = 11.10 (s, 1H), 9.23-9.11 (m, 3H), 8.34 (d, J = 8.0 Hz, 1H), 8.10 (s, 1H), 7.55 (s, 1H), 7.44 (d,J = 8.0 Hz, 1H), 7.29-7.01 (m, 3H), 5.76 (s, 1H), 5.28 (1, 1H), 3.82-3.77 (m,4H), 3.68-3.55 (m, 4H), 2.44 (s, 3H), 2.44-2.31 (m, 4H), 2.13 (s, 3H), 2.03-1.91 (m, 2H) ppm; HRMS (ESI): m/z [M+H]+. C34H35ClN11O5S计算值744.2226,实测值744.2238。
实施例2
Bcr-abl-PROTAC-020的制备
2-((6-(4-(2-(2-(叔丁基)-4-(2,4-二氧-3,4-二氢嘧啶-1(2H)-基)苯氧基)乙基)哌嗪-1-基)-2-甲基嘧啶-4-基)氨基)-N-(2-氯-6-甲基苯基)噻唑-5-甲酰胺合成路线:
步骤一:称取原料2-(哌嗪-1-基)乙醇(2.0 g, 15.4 mmol)于100 mL单颈瓶中,加入无水四氢呋喃(40 mL)溶解上述原料,继而加入氯化亚砜(3.67 g, 30.8 mmol)后,升温至回流反应过夜。次日,停止反应待反应液降至室温后,真空浓缩得到中间体3-1,无需纯化即可用于下一步反应。(2.28 g, 100%)HRMS (ESI): m/z [M+H]+. C6H14ClN2计算值149.0840,实测值149.0851。
步骤二:将上一步产物(1.0 g, 6.76 mmol)溶于正丁醇(30 mL)中,加入达沙替尼中间体原料(3.2 g, 8.10 mmol),DIPEA(1.74 g, 13.52 mmol)后,升温至回流反应过夜,TLC监测反应完全后,加水并使用二氯甲烷萃取2-3次,合并有机层,水洗,饱和食盐水洗,真空浓缩后将残余物拌入硅胶制砂,柱层析(DCM:MeOH = 40:1)得到中间体3-2。(1.1 g,32.2%)HRMS (ESI): m/z [M+H]+. C22H26Cl2N7OS计算值506.1291,实测值506.1299。
步骤三:称取中间体3-2(1.0 g, 1.98 mmol)于100 mL单颈瓶中,室温下加入2-叔丁基-4-碘苯酚(0.65 g, 2.38 mmol),无水碳酸钾(0.55 g, 3.96 mmol)及无水DMF(20mL),升温至80 oC搅拌反应过夜,TLC监测反应完全后,加水并使用二氯甲烷萃取2-3次,合并有机层,水洗,饱和食盐水洗,真空浓缩后将残余物拌入硅胶制砂,柱层析(DCM:MeOH =30:1)得到中间体3-3。(0.6 g, 40.7%)HRMS (ESI): m/z [M+H]+.C32H38ClIN7O2S计算值746.1535,实测值746.1544。
步骤四:称取中间体3-3(0.5 g, 0.67 mmol)于50 mL单颈瓶中,室温下加入尿嘧啶(0.11 g, 1.0 mmol),碘化亚铜(0.2 g, 1.0 mmol),L-脯氨酸(0.12 g, 1.0 mmol),磷酸三钾(0.21 g, 1.0 mmol),无水DMSO(10 mL),N2气保护下60 oC搅拌反应过夜,TLC监测反应完全后,加水并使用二氯甲烷萃取2-3次,合并有机层,水洗,饱和食盐水洗,真空浓缩后将残余物拌入硅胶制砂,柱层析(DCM:MeOH = 20:1)得到目标产物bcr-abl-PROTAC-020纯品。(0.1 g, 20.5%)1HNMR(400 MHz, CDCl3): δ = 10.60 (s, 1H), 9.27-9.15 (m, 2H),8.68 (s, 1H), 8.15 (s, 1H), 7.47 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 8.0 Hz,1H), 7.02-6.86 (m, 3H), 5.81 (s, 1H), 5.33 (1, 1H), 4.15 (d, J = 8.0 Hz, 2H),3.62-3.54 (m, 4H), 3.44-3.32 (m, 4H), 2.76-2.54 (m, 2H), 2.47 (s, 3H), 2.15(s, 3H), 1.38 (s, 9H) ppm;HRMS (ESI): m/z [M+H]+. C36H41ClN9O4S计算值730.2685,实测值730.2688。
实施例3
本发明公开了一种新型PROTAC化合物及其在癌症药物中的用途,本发明中构建的蛋白降解靶向嵌合体(PROTAC)可使靶蛋白泛素化,通过泛素-蛋白酶体降解靶蛋白,避免由于靶点突变产生的耐药性。本发明为构建蛋白降解靶向嵌合体提供了新选择,构建的蛋白降解靶向嵌合体在制备预防和/或治疗免疫疾病和癌症药物中有良好的应用,具体的应用实施例包括但不限于以下:
以达沙布韦(Dasabuvir)的衍生物/结构类似物(式Ⅳ,式Ⅴ)为E3连接酶Cereblon(CRBN)的新型配体,临床用药达沙替尼 (Dasatinib)等为BCR-ABL靶蛋白配体,构建得到了靶向癌蛋白BCR-ABL的PROTAC化合物Bcr-abl-PROTAC-No.14(式Ⅰ),Bcr-abl- PROTAC-No.20(式Ⅱ),两种化合物均可有效降解K562癌细胞中的BCR-ABL癌蛋白,同时显著降低K562癌细胞的细胞增殖。
具体地,内源性表达BCR-ABL的白血病细胞系K562进行常规细胞培养(培养基为含有10%FBS的RPMI1640培养基,Giboco公司所产),分别给与DMSO(对照), 阳性对照药(GMB-475),Bcr-abl-PROTAC-014以及Bcr-abl-PROTAC-020化合物处理24小时(将药物溶解于RPMI1640培养基中,终浓度为40 μM)。目标蛋白BCR-ABL蛋白以及内参蛋白beta-Actin 通过常规蛋白质印记法Western Blot方法检测,如图1,图2所示,实验中所用目标蛋白和内参蛋白的识别抗体分别为ab187831及ab8227,均来自Abcam公司。此外,K562癌细胞系用DMSO(对照),阳性对照药(GMB-475),Bcr-abl-PROTAC-014以及Bcr-abl-PROTAC-020化合物处理48小时(同上,将药物溶解于RPMI1640培养基中,终浓度为10 μM),通过Cell-Titer Glo方法检测细胞活力,如图3,图4所示。由上述实验结果可知, Bcr-abl-PROTAC-014以及Bcr-abl-PROTAC-020化合物对K562癌细胞均具有较好的抑制作用。由此,以上实验提示Bcr-abl-PROTAC-014以及Bcr-abl-PROTAC-020化合物在抗癌症药性方面有独特的优势,是一个有开发前景的潜在药物。
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US20180215731A1 (en) * | 2017-01-31 | 2018-08-02 | Arvinas, Inc. | Cereblon ligands and bifunctional compounds comprising the same |
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US20180215731A1 (en) * | 2017-01-31 | 2018-08-02 | Arvinas, Inc. | Cereblon ligands and bifunctional compounds comprising the same |
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ASHTON C LAI ET AL: """Modular PROTAC Design for the Degradation of Oncogenic BCR-ABL"" * |
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