一类靶向降解雄激素受体的双功能嵌合体杂环化合物及其
用途
技术领域
本发明属于医药领域,具体而言,本发明涉及一类靶向降解雄激素受体的双功能嵌合体杂环化合物及其用途。
背景技术
在全球人口不断增长和老龄化的情况下,前列腺癌发病率持续地增长,目前其主要的治疗手段为雄激素剥夺治疗。雄激素受体(androgen receptor,AR)属于核受体家族,是一类配体依赖的转录因子。AR信号通路的异常调节对前列腺癌的发生、发展有重要作用,研究表明去势抵抗型前列腺癌(castration-resistant prostate cancer,CRPC)仍然依赖AR的作用。雄激素受体包含918个氨基酸,与其他核受体具有相似的结构和功能,它由三个重要的结构域组成,分别是DNA结合域(DNA binding domain,DBD)、配体结合域(ligandbinding domain,LBD)和氮端结合域(N-terminal domain,NTD),DBD和LBD之间通过一个铰链区(Hinge)相连。存在于AR碳端的LBD是AR与配体结合的位点,决定了配体与AR结合的特异性,配体与LBD结合从而激活AR。在AR中已确定了两个转录激活功能区,即NTD结构域中的激活功能区1(activation function 1,AF1)和LBD结构域中高度保守的疏水口袋激活功能区2(AF2)。2010年以前,以多西紫杉醇为基础的化疗是唯一能延长转移性CRPC患者生存期的治疗方法。
蛋白降解靶向嵌合体(PROTACs)作为能够诱导靶蛋白降解的小分子受到了广泛关注。PROTACs作为双功能分子,包括一个能够与靶蛋白(protein of interest,POI)结合的小分子化合物,在其合适位置上引入连接基团,再与一个能够与泛素蛋白酶结合的小分子化合物连接。得到的小分子探针可以同时与靶蛋白和泛素蛋白酶结合,从而促使靶蛋白泛素化,被多重泛素化的蛋白可以被蛋白酶体识别并降解。
利用PROTACs策略,制备出能够靶向识别/结合雄激素受体的蛋白降解靶向嵌合体,能够通过细胞内泛素-蛋白酶体降解系统而调控雄激素受体的水平,诱导雄激素受体降解,从而达到治疗前列腺癌等受雄激素受体调控的相关疾病的效果。
所以,研发出一种能够与雄激素受体靶向结合、有效降解雄激素受体的双功能嵌合体分子,在治疗受雄激素受体调控的相关疾病上具有良好的应用前景。
发明内容
本发明的目的在于提供一种与雄激素受体靶向结合能力更强、降解雄激素受体的活性更高的蛋白降解靶向嵌合体。
本发明提供了式(I)所示的化合物、或其同位素化合物、或其光学异构体、或其互变异构体、或其药学上可接受的盐、或其前药、或其溶剂合物:
其中,ARB为雄激素受体识别/结合部分,L为链接部分,U为泛素蛋白酶识别/结合部分;这三个部分通过化学键相连接;
其中,上述ARB选自式(I-A)所示的结构:
其中,W1选自取代或未取代的芳基或杂芳基,所述W1上的取代基各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;
Y1,Y2,Y3,Y4各自独立地选自一个键,O,S,NR1,CR2R3,C=O,C=S,SO或SO2;上述R1,R2,R3分别独立地选自H,C1-6烷基或其卤代物或氘代物,含0-2个杂原子的3-8元环烷基或杂环基,或者R2与R3链接起来形成含0-2个杂原子的3-8元环;
Q选自被0~6个Rq取代的饱和环烷基、饱和杂环基、或含0-4个杂原子的芳基或杂芳基,所述Rq各自独立地选自H,D,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者2个取代基连接起来构成含有0-2个杂原子的3-8元环;
W2选自一个键,或被0~4个R4取代的下述基团:烯基、炔基、C1-6烷基、C1-6烷氧基、单环烷基、单杂环基、芳基、杂芳基、桥环烷基、杂桥环基、螺环烷基、杂螺环基、稠环烷基、杂稠环基;
上述R
4各自独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF
3,C
1-6烷基或其卤代物或其氘代物,C
3-6环烷基,C
1-6烷氧基或其卤代物或其氘代物,C
1-6烷胺基,C
2-6炔基,C
2-6烯基,或R
4选自
R
1c选自O或S;
或,
ARB选自式(I-B)所示的结构:
其中,W1选自取代或未取代的芳基或杂芳基,所述W1上的取代基各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;
Y1,Y5,Y6各自独立地选自一个键,O,S,NR1,CR2R3,C=O,C=S,SO,SO2;上述R1,R2,R3分别独立地选自H,C1-6烷基或其卤代物或氘代物,含0-2个杂原子的3-8元环烷基或杂环基,或者R2与R3链接起来形成含0-2个杂原子的3-8元环;
Q选自被0~6个Rq取代的饱和环烷基、饱和杂环基、或含0-4个杂原子的芳基或杂芳基,所述Rq各自独立地选自H,D,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者2个取代基连接起来构成含有0-2个杂原子的3-8元环;
W2选自一个键,或被0~3个R4取代的烯基、炔基、C1-6烷基、C1-6烷氧基、单环烷基、单杂环基、芳基、杂芳基、桥环烷基、杂桥环基、螺环烷基、杂螺环基、稠环烷基、杂稠环基;
其中R4各自独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6炔基,C2-6烯基;
或,
ARB选自式(I-C)所示的结构:
其中,W1选自取代或未取代的芳基或杂芳基,所述W1上的取代基各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;
Y1选自一个键,O,S,NR1,CR2R3,C=O,C=S,SO,SO2;
Y7选自N,CR2;Y8选自一个键,O,S,NR1,CR2R3,C=O,C=S,SO,SO2;
Y7和W2相连接,与Y8一起形成4-7元环,所述环被0~4个氘、卤素取代;
上述R1,R2,R3分别独立地选自H,C1-6烷基或其卤代物或其氘代物,含0-2个杂原子的3-8元环烷基或杂环基,或者R2与R3链接起来形成含0-2个杂原子的3-8元环;
Q选自被0~6个Rq取代的饱和环烷基、饱和杂环基、或含0-4个杂原子的芳基或杂芳基,所述Rq各自独立地选自H,D,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者2个取代基连接起来构成含有0-2个杂原子的3-8元环;
W2选自一个键,或被0~3个R4取代的烯基、炔基、C1-6烷基、C1-6烷氧基、单环烷基、单杂环基、芳基、杂芳基、桥环烷基、杂桥环基、螺环烷基、杂螺环基、稠环烷基、杂稠环基;
其中R4各自独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6炔基,C2-6烯基;
或,
ARB选自式(I-D)所示的结构:
其中,W1选自取代或未取代的芳基或杂芳基,所述W1上的取代基各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;
Y9,Y10,Y11分别独立地选自CH,O,S;
Y12选自一个键,或CO,CO2,O,S,NR1e,NR1eCO,NR1eSO2;所述R1e选自H,C1-6烷基或其卤代物或其氘代物,含0-2个杂原子的3-8元环烷基或杂环基;
Rq各自独立地选自H,D,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者2个Rq连接起来构成含有0-2个杂原子的3-8元环;
W2选自一个键,或被0~3个R4取代的烯基、炔基、C1-6烷基、C1-6烷氧基、单环烷基、单杂环基、芳基、杂芳基、桥环烷基、杂桥环基、螺环烷基、杂螺环基、稠环烷基、杂稠环基;R4各自独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6炔基,C2-6烯基。
进一步地,
所述式(I-A)、式(I-B)或式(I-C)中的
结构如下述式(II-A)、式(IV-B)、(IV-C)、(IV-D)、(IV-E)、(IV-F)、(IV-G)、或(IV-H)所示:
其中,Rq各自独立地选自H,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者2个Rq连接起来构成含有0-2个杂原子的3-8元环;
a,b,c,d分别独立地选自0-3的整数。
进一步地,所述式(I-A)、式(I-B)或式(I-C)中的
结构如下述式(III-A)或式(III-B)或式(III-C)或式(III-D)所示:
或,所述式(I-A)、式(I-B)或式(I-C)中的
结构如下述式(III-E)、式(III-F)、式(III-G)、式(III-H)、式(III-I)、式(III-J)、式(III-K)、式(III-L)、式(III-M)或式(III-N)所示:
其中,Rw1,Rw2,Rw3,Rw4,Rw5各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;所述卤素优选为氯或溴,所述C1-6烷基优选为甲基,所述C1-6烷氧基优选为甲氧基或乙氧基;
Rq各自独立地选自H,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者任意两个Rq连接起来构成含有0-2个杂原子的3-8元环。
进一步地,:
进一步地,所述式(I-D)结构如下述式(V-D)所示:
其中,W1、W2、Y12、Rq如上述式(I-D)中所述。
进一步地,所述ARB选自如下式的结构:
其中,Rw1,Rw2,Rw3,Rw4,Rw5各自独立地选自卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,杂环基,CF3,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6烯基或C2-6炔基;所述卤素优选为氯或溴,所述C1-6烷基优选为甲基,所述C1-6烷氧基优选为甲氧基或乙氧基;
Rw6选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,C1-6烷基或其卤代物或其氘代物,C3-6环烷基,C1-6烷氧基或其卤代物或其氘代物,C1-6烷胺基,C2-6炔基,C2-6烯基;
Rq各自独立地选自H,OH,卤素,C1-6烷基或其卤代物,C1-6烷氧基或其卤代物,或者任意两个Rq连接起来构成含有0-2个杂原子的3-8元环;
Y12选自一个键,或CO,CO2,O,S,NR1e,NR1eCO,NR1eSO2;所述R1e选自H,C1-6烷基或其卤代物或其氘代物。
进一步地,所述ARB选自如下式的结构:
或,所述ARB选自如下式的结构:
进一步地,所述L选自如下式(VIII-A)的结构:
其中:L1,L2,L3,L4,L5,L6分别独立地选自无,一个键,O,S,NRL1,CRL2RL3,C=O,C=S,SO,SO2,取代或未取代的烯基,取代或未取代的炔基,取代或未取代的单环烷基,取代或未取代的单杂环基,取代或未取代的芳基,取代或未取代的杂芳基,取代或未取代的桥环烷基,取代或未取代的杂桥环基,取代或未取代的螺环烷基,取代或未取代的杂螺环基,取代或未取代的稠环烷基,取代或未取代的杂稠环基;
上述取代基选自C1-6烷基、-L7-OH、卤素,L7选自0~6个亚甲基;
RL1,RL2,RL3分别独立地选自H,C1-6烷基或其卤代物或其氘代物,含0-2个杂原子的3-8元环烷基或杂环基,或者RL2,RL3链接起来形成含0-2个杂原子的3-8元环;
a,b,c,d,e,f分别独立地选自0-5的整数;
L1和L6可以分别自由地和ARB或U连接;
或者,所述L选自式(VIII-B)所示的结构:
其中环A,环B各自独立地选自卤代或未卤代的如下结构:
X0选自:无,O,S,SO,SO2,NRX1,CRX1RX2;RX1,RX2分别独立地选自H,卤素,C1-C6烷基,C3-C6环烷基,卤素或羟基或氨基取代的C1-C6烷基,C1-C6烷基主链中的碳原子被氧或氮原子替换后得到的基团,杂环基,芳基,羟基,氨基,或者RX1和RX2相连接形成3-7元环;
环A和环B可以分别自由地和ARB或U连接。
进一步地,所述式(VIII-A)选自如下所示的结构:
其中,X选自H或卤素,m、n各自独立地选自0~5的整数。
进一步地,所述L选自如下所示的结构:
优选地,所述L选自如下结构:
进一步地,所述U选自如下式(X-A)的结构:
其中:T,Y分别独立地选自一个键,O,S,NRT1或CRT2RT3;
V,J分别独立地选自一个键,C=O,-SO-,-SO2-或CR2R3;
RT1,RT2,RT3分别独立地选自H,C1-6烷基或其卤代物或其氘代物,含0-2个杂原子的3-8元环烷基或杂环基,或者RT2与RT3链接起来形成含0-2个杂原子的3-8元环;
Rv选自H,C1-6烷基或其卤代物或其氘代物,含0-3个杂原子的环烷基或杂环基、或其卤代物,或者Rx与Ry链接起来形成含0-2个杂原子的3-8元环;
g,h分别独立地选自0-3的整数,且g和h不同时为0;
Z选自H,羟基,氨基,C1-6烷基,C3-6环烷基,氧或卤素取代的C1-6烷基,-ORZ1,-NRZ1RZ2,-CORZ3,-CO2RZ3,-OCORZ3,-NHCORZ3,-CONHRZ3,-SO2RZ3;RZ1,RZ2分别独立地选自H,C1-6烷基或其卤代物或其氘代物,含0-2个杂原子的3-8元环烷基或杂环基;
上述RZ3选自取代或未取代的C1-6烷基,取代或未取代的C3-6环烷基,取代或未取代的C3-6杂环基,取代或未取代的芳基,取代或未取代的杂芳基;所述RZ3上的取代基选自卤素、C1-3烷基;
Rx,Ry分别独立地选自H,C1-6烷基,卤代C1-6烷基,被含杂原子的基团取代的C1-6烷基,-Ly-OH,含0-3个杂原子的环烷基或杂环基、或其卤代物,或者Rx与Ry链接起来形成含0-2个杂原子的3-8元环;其中,Ly选自0~5个亚甲基;
W4、W5分别独立地选自被0~3个取代基取代的芳基、杂芳基,所述取代基各自独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基,C3-6环烷基,C1-6烷氧基,C1-6烷胺基,C2-6烯基,C2-6炔基;
或,
所述U选自如下式(X-B)的结构:
其中,M选自O,S,NR
m;其中R
m选自H,C
1-6烷基,C
3-6环烷基,C
3-6杂环基,
上述Rm1选自H,C1-6烷基,C3-6环烷基;Xm选自无,O,S,NRm3;
R
m2,R
m3分别独立地选自H,C
1-6烷基,C
3-6环烷基,C
3-6杂环基,
上述i选自0~12的整数,R
m4选自H、C
1-6烷基,L
m选自0~5个亚甲基,M
a选自N、CH,M
b选自O、S、CH
2、NH;
E、F各自独立地选自CO,CS,NR
e1,O,S,SO
2,CH
2,CD
2,CR
e2R
e3,
R
e1,R
e2,R
e3分别独立地选自C
1-6烷基,H,卤素,羟基,氨基;
Y15,Y13,Y14分别独立地选自H,O,S,C1-3烷基;
j,k分别独立地选自0-3的整数,且j,k不同时为0;
G1,G2,G3,G4分别独立地选自O,S,N,CRg1,CRg2,CRg3,CRg4,其中Rg1,Rg2,Rg3,Rg4分别独立地选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基,C3-6环烷基,C1-6烷氧基,C1-6烷胺基,C2-6烯基,C2-6炔基;
Ru1选自H,C1-C6烷基;
或,
所述U选自如下式(X-C)的结构:
进一步地,所述式(X-A)选自如下式(XI-A)所示的结构:
其中,Rv、Z、g、h、Rx、Ry、W4、W5的选择范围与上述式(II-A)中相同;
或,
所述式(X-B)中
选自如下式(XI-B)、(XI-C)、(XI-D)、(XI-E)、或(XI-F)所示的结构:
其中,G1、G2、G3、G4的选择范围与上述式(X-B)中相同。
进一步地,所述式(XI-A)选自如下式(XII-A)所示的结构:
其中,Rw7选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基,C3-6环烷基,C1-6烷氧基,C1-6烷胺基,C2-6烯基,C2-6炔基;
M1,M2,M3,M4分别独立地选自O,S,N R12,C(R12)2,其中R12选自H,卤素,羟基,氨基,巯基,砜基,亚砜基,硝基,氰基,CF3,杂环基,C1-6烷基,C3-6环烷基,C1-6烷氧基,C1-6烷胺基,C2-6烯基,C2-6炔基,
Rv、Z、Rx、Ry的选择范围与上述式(XI-A)中相同。
进一步地,所述式(XI-A)中W5选自下列结构:
进一步地,所述U选自下列结构:
进一步地,所述化合物选自以下化合物之一:
本发明还提供了上所述的化合物、或其同位素化合物、或其光学异构体、或其互变异构体、或其药学上可接受的盐、或其前药、或其溶剂合物在制备雄激素受体的蛋白降解靶向嵌合体中的用途。
进一步地,所述蛋白降解靶向嵌合体能够靶向识别和/或结合雄激素受体。
进一步地,所述蛋白降解靶向嵌合体能够降解和/或下调雄激素受体。
进一步地,所述蛋白降解靶向嵌合体为治疗受雄激素受体调控的相关疾病药物的活性成分。
进一步地,所述疾病选自前列腺癌、乳腺癌、肯尼迪氏病。
本发明还提供了一种治疗受雄激素受体调控的相关疾病的药物,所述药物是以上述的化合物、或其同位素化合物、或其光学异构体、或其互变异构体、或其药学上可接受的盐、或其前药、或其溶剂合物为活性成分,加上药学上可接受的辅料制得的制剂。
通过实验证明,本发明提供的式(I)所示的化合物能够靶向降解前列腺癌细胞中的雄激素受体,并且抑制前列腺癌细胞的增殖,同时还显示了良好的代谢稳定性和药代动力学性质。本发明化合物在制备雄激素受体的蛋白降解靶向嵌合体,以及制备治疗受雄激素受体调控的相关疾病(包括前列腺癌、乳腺癌、肯尼迪氏病)的药物中具有良好的应用前景。
关于本发明的使用术语的定义:除非另有说明,本文中基团或者术语提供的初始定义适用于整篇说明书的该基团或者术语;对于本文没有具体定义的术语,应该根据公开内容和上下文,给出本领域技术人员能够给予它们的含义。
碳氢基团中碳原子含量的最小值和最大值通过前缀表示,例如,前缀Ca~b烷基表示任何含“a”至“b”个碳原子的烷基。例如,C1~6烷基是指包含1~6个碳原子的直链或支链的烷基。
本文“取代”是指分子中的1个、2个或多个氢原子被其它不同的原子或分子所替换,包括该分子中同位原子或异位原子上的1个、2个或多个取代。
本文碳氢基团中碳原子含量的最小值和最大值通过前缀表示,例如,C1-6烷基表示任何含1-6个碳原子的直链或支链烷基;C1-6烷氧基表示任何含1-6个碳原子的直链或支链烷氧基。
本发明中,“芳基”指具有共轭的π电子体系的全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团,例如苯基和萘基。所述芳基环可以稠合于其它环状基团(包括饱和和不饱和环),但不能含有杂原子如氮,氧,或硫,同时连接母体的点必须在具有共轭的π电子体系的环上的碳原子上。芳基可以是取代的或未取代的。
“杂芳基”指包含一个到多个杂原子的杂芳族基团。这里所指的杂原子包括氧、硫和氮。例如呋喃基、噻吩基、吡啶基、吡唑基、吡咯基、N-烷基吡咯基、嘧啶基、吡嗪基、咪唑基、四唑基等。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上,其中与母体结构连接在一起的环为杂芳基环。杂芳基可以是任选取代的或未取代的。
“烷基”是烷烃分子中少掉一个氢原子而成的烃基,例如甲基-CH3,乙基-CH3CH2等。
“烷胺基”是烷基中的一个或多个氢原子被胺基取代后得到的基团。
“炔基”是指具有至少一个碳-碳三键的脂肪族碳氢基团。所述的炔基可以是直链或支链的。当炔基前具有碳原子数限定时,例如,“C2-6炔基”指具有2-6个碳原子的直链或支链炔基。
“烯基”是指具有至少一个碳-碳双键的脂肪族碳氢基团。所述的烯基可以是直链或支链的。当烯基前具有碳原子数限定时,例如,“C2-6烯基”指具有2-6个碳原子的直链或支链烯基。
“取代或未取代的烯基”指烯基可以是被取代的,也可以没有取代基的。
“环烷基”指饱和或不饱和的环状烃取代基;环状烃可以是单环也可以是多环。例如,“3-8元环烷基”指碳原子数为3~8的环烷基。
“饱和环烷基”指饱和的环烷基。
“单环烷基”指该环烷基是单环的。
“桥环烷基”指多环的环烷基,且该多环的环烷基中有两个环共用两个不相邻的碳原子。
“螺环烷基”指多环的环烷基,且该多环的环烷基中有两个环共用一个碳原子。
“稠环烷基”指多环的环烷基,且该多环的环烷基中有两个环共用两个相邻的碳原子。
“杂环基”指饱和或不饱和的环状烃取代基;环状烃可以是单环也可以是多环,且携带至少一个环杂原子(包括但不限于O、S或N)。例如,“3~8元杂环基”指碳原子数为3~8的杂环基。
“饱和杂环基”指饱和的杂环基。
“单杂环基”指该杂环基是单环的。
“杂桥环基”多环的杂环基,且该多环的杂环基中有两个环共用两个不相邻的碳原子或杂原子。
“杂螺环基”多环的杂环基,且该多环的杂环基中有两个环共用一个碳原子或杂原子。
“杂稠环基”多环的杂环基,且该多环的杂环基中有两个环共用两个相邻的碳原子或杂原子。
卤素为氟、氯、溴或碘。
“同位素化合物”指化合物中的一个或两个以上的原子被其对应的同位素替换后得到的化合物。比如化合物中的一个或两个以上的氢(H)被氘(D)或氚(T)替换后得到的化合物;比如化合物中的一个或两个以上的碳12被碳11或碳13替换后得到的化合物。
“药学上可接受的”是指某载体、运载物、稀释剂、辅料,和/或所形成的盐通常在化学上或物理上与构成某药物剂型的其它成分相兼容,并在生理上与受体相兼容。
“盐”是将化合物或其立体异构体,与无机和/或有机酸和/或碱形成的酸式和/或碱式盐,也包括两性离子盐(内盐),还包括季铵盐,例如烷基铵盐。这些盐可以是在化合物的最后分离和纯化中直接得到。也可以是通过将化合物,或其立体异构体,与一定数量的酸或碱适当(例如等当量)进行混合而得到。这些盐可能在溶液中形成沉淀而以过滤方法收集,或在溶剂蒸发后回收而得到,或在水介质中反应后冷冻干燥制得。本发明中所述盐可以是化合物的盐酸盐、硫酸盐、枸橼酸盐、苯磺酸盐、氢溴酸盐、氢氟酸盐、磷酸盐、乙酸盐、丙酸盐、丁二酸盐、草酸盐、苹果酸盐、琥珀酸盐、富马酸盐、马来酸盐、酒石酸盐或三氟乙酸盐。
“其溶剂合物”指本发明化合物与溶剂形成溶剂合物,其中,所述溶剂包括(但并不限于):水、乙醇、甲醇、异丙醇、丙二醇、四氢呋喃、二氯甲烷。
本发明的化合物中,ARB为雄激素受体识别/结合部分,在化合物中发挥雄激素受体的配体的作用。
本发明中“选自一个键”指所述基团或原子为无,其两端的连接位点直接相连,比如式(I-A)
中,Y
3选自一个键时,所述结构为
本发明式(XII-A)结构中的
表示由CH、M
1、M
2、M
3、M
4组成的,具有共轭结构的五元环。
本发明式(VIII-A)中“L1和L6可以分别自由地和ARB或U连接”表示L1与ARB连接、同时L6与U连接,或者L1与U连接、同时L6与ARB连接。
同样的,本发明式(VIII-B)中“环A和环B可以分别自由地和ARB或U连接”表示环A与ARB连接、同时环B与U连接,或者环A与U连接、同时环B与ARB连接。
本发明中,“D”表示氘。
本发明的具体化合物命名方式有多种:(1)数字编号,比如化合物“315”、“3”;(2)化合物命名,如(3R,5S)-1-((S)-2-(2-((5-((4-(3-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基))苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯。虽然命名方式不同,但是本发明的各个具体化合物根据其结构可唯一确定。
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为本发明化合物99在不同浓度下的蛋白免疫印迹实验结果。
具体实施方式
本发明具体实施方式中使用的原料、设备均为已知产品,通过购买市售产品获得。
首先,合成中间体:
SM-A-1:4-((1r,3r)-3-(2-溴-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈
1.化合物3-(甲氧羰基)-2-甲基吡啶-1-氧代物
将化合物2-甲基烟酸甲酯(30.0g,199.0mmol)溶入二氯甲烷(500mL),然后分批加入间氯过氧苯甲酸(68.7g,398.0mmol)。室温搅拌过夜,过滤,滤饼用二氯甲烷洗涤。滤液加入5%亚硫酸钠溶液(200mL)搅拌30分钟,分液,并用二氯甲烷(100mL×3)萃取,滤液合并,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化(二氯甲烷:甲醇=100:1至20:1),得到白色固体化合物3-(甲氧羰基)-2-甲基吡啶-1-氧代物(18g,107.8mmol),收率:54%。MS:calcd.for C8H9NO3[M+H]+:168.0;found:168.1.
2.化合物6-氯-2-甲基烟酸甲酯
将化合物3-(甲氧羰基)-2-甲基吡啶-1-氧代物(36.5g,218.6mmol)在冰浴下分批加入到三氯氧磷(300mL)中。随后缓慢升温回流反应3小时。反应液减压除去大部分溶剂,用乙酸乙酯(500mL)稀释,然后用10%Na2CO3水溶液洗涤(100mL×3),再用饱和食盐水洗涤(100mL×3)。有机层用无水硫酸钠干燥,旋干,硅胶柱层析(石油醚:乙酸乙酯=10:1至5:1)纯化得到白色固体6-氯-2-甲基烟酸甲酯(8g,43.2mmol)。收率:20%。MS:calcd.forC8H8ClNO2[M+H]+:186.0;found:186.1.
3.化合物6-溴-2-甲基烟酸甲酯
将化合物6-氯-2-甲基烟酸甲酯(8.0g,43.2mmol)溶入乙腈(80mL),然后加入三甲基溴硅烷(19.8g,129.7mmol),反应液加热回流过夜。反应液减压除去溶剂,残渣用硅胶柱层析(石油醚:乙酸乙酯=30:1至10:1)纯化得到黄色固体6-溴-2-甲基烟酸甲酯(7.0g,30.6mmol)。收率:71%。MS:calcd.for C8H8BrNO2[M+H]+:230.0;found:230.1.
4.化合物2-(溴甲基)-6-溴烟酸甲酯
将化合物6-溴-2-甲基烟酸甲酯(7.0g,30.6mmol)和N-溴代丁二酰亚胺(8.16g,45.9mmol)加入到四氯化碳(100mL),随后加入偶氮二异丁腈(566mg,3.06mmol),反应液回流过夜。反应液减压除去溶剂,残渣用硅胶柱层析(石油醚:乙酸乙酯=30:1至10:1)得到2-(溴甲基)-6-溴烟酸甲酯(A-1-5)粗品。
粗品溶入二氯甲烷(60mL),加入N,N-二异丙基乙胺(2.36g,18.3mmol),冰浴滴入亚磷酸二乙酯(1.21g,8.76mmol),室温搅拌过夜。反应液加水(100mL)稀释,二氯甲烷萃取(40mL×3),饱和食盐水洗涤(50mL×3),有机相合并加入无水硫酸钠干燥,旋干,硅胶柱层析(石油醚:乙酸乙酯=30:1至10:1)纯化得到白色固体2-(溴甲基)-6-溴烟酸甲酯(6.0g,19.5mmol),收率:64%。MS:calcd.for C8H7Br2NO2[M+H]+:307.9;found:308.1.
5.化合物3-羰基-2,2,4,4-四甲基环丁酮肟
将盐酸羟胺(26.0g,374.5mmol)溶入水(40mL)和乙醇(250mL)中,然后加入1,3-四甲基环丁二酮(50g,356.7mmol)和醋酸钠(29.3g,356.7mmol)。反应液加热回流2小时。旋蒸除去乙醇和水。残渣加入甲苯(300mL)后回流3小时,然后热过滤,滤饼用甲苯(100mL)洗涤。滤液旋干得到白色固体3-羰基-2,2,4,4-四甲基环丁酮肟(32.0g,206.5mmol),收率:59%。
MS:calcd.for C8H13NO2[M+H]+:156.1;found:156.1.
6.化合物3-羟基-2,2,4,4-四甲基环丁酮肟
将3-羰基-2,2,4,4-四甲基环丁酮肟(110.9g,714.6mmol)溶入异丙醇(715mL),然后分批加入硼氢化钠(18.9g,500.2mmol)。反应液室温搅拌过夜。反应液控制在5℃用氢氧化钠(2000mL)淬灭。然后用乙酸乙酯(700mL×3)萃取,有机相合并用硫酸钠干燥,旋干得到白色固体3-羟基-2,2,4,4-四甲基环丁酮肟(100.8g,642.0mmol),收率:90%。
MS:calcd.for C8H15NO2[M+H]+:158.1;found:158.1.
7.化合物3-氨基四甲基环丁酮
将3-羟基-2,2,4,4-四甲基环丁酮肟(A-1-7)(35.1g,223.3mmol)溶入四氢呋喃(300mL),然后在氮气保护下加入镍铝合金(76.5g,893.3mmol)。反应液回流30分钟后,加入15%氢氧化钠(300mL)并保持回流状态,加完后继续回流2小时。反应完成后过滤,滤饼用四氢呋喃(100mL×3)洗涤,滤液用乙酸乙酯(200mL×3)萃取,有机相合并用饱和食盐水(200mL×3)洗涤,硫酸钠干燥,旋干得到淡黄色固体3-氨基四甲基环丁酮(24.5g,171.3mmol),收率:77%。
MS:calcd.for C8H17NO[M+H]+:144.1;found:144.1.
8.化合物3-羟基-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯
将3-氨基四甲基环丁酮(26.9g,188.1mmol)溶入二氯甲烷(500mL),然后加入二碳酸二叔丁酯(41.4g,190.0mmol)和三乙胺(38.0g,376.2mmol)。反应液室温搅拌过夜。反应液用水(300mL)稀释,二氯甲烷(100mL×3)萃取。有机相合并用饱和食盐水(200mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至1:1)纯化得到淡黄色固体3-羟基-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯(30.1g,123.9mmol),收率:66%。
MS:calcd.for C8H17NO[M+H]+:244.2;found:244.2.
9.化合物(1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯
将3-羟基-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯(25.0g,102.9mmol)溶入四氢呋喃(250mL)中,冰浴下分批加入氢化钠(60%在矿物油中)(8.23g,205.8mmol)并在冰浴中搅拌30分钟,然后将2-氯-4-氟苯腈(17.6g,113.2mmol)溶于四氢呋喃(50mL)中,慢慢滴加到反应中。反应液加热到60℃反应3小时。反应液用水(300mL)淬灭,乙酸乙酯(100mL×3)萃取。有机相合并用饱和食盐水(200mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=50:1至30:1)纯化得到白色固体(1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯(15.0g,39.7mmol),收率:39%。MS:calcd.for C20H27ClN2O3[M+H]+:379.2;found:279.2.
10.化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈三氟乙酸盐
将(1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基氨基甲酸叔丁酯(5.0g,13.2mmol)溶入二氯甲烷(50mL)中,在冰浴下加入三氟乙酸(50mL)。反应液在室温搅拌1小时,旋干得到4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈三氟乙酸盐粗品,直接用于下一步反应。MS:calcd.for C17H19ClF3N2O2[M+H]+:376.1;found:279.1.
11.化合物4-((1r,3r)-3-(2-溴-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈
将4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈三氟乙酸盐(13.2mmol,粗品来源于上一步)溶入乙腈(240mL)中,然后加入N,N-二异丙基乙胺(8.51g,66.0mmol)和化合物2-(溴甲基)-6-溴烟酸甲酯(4.0g,13.2mmol)。反应也加热回流过夜。反应也旋干后,加入甲苯(50mL)回流过夜。反应液冷却过滤,滤饼用甲苯洗涤,滤饼干燥得到目标化合物为白色固体(3.95g,8.3mmol),两步收率:63%。MS:calcd.for C22H21BrClN3O2[M+H]+:474.0;found:474.0.
SM-A-2,SM-A-3,SM-A-4,SM-A-5用类似于SM-A-1的方法制备得到。
SM-A-2:2-氯-4-((1r,3r)-3-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(527mg,1.89mmol)、碳酸钾(392mg,2.84mmol)和10mL DMF依次加入,氮气保护,搅拌均匀后滴入2-(溴甲基)-6-氯烟酸甲酯(500mg,1.89mmol)的DMF溶液,室温搅拌2h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到中间体。中间体加入DIEA(734mg,1.48mmol)和5mL 1,4二氧六环,100℃反应2d,TLC确定反应终点硅胶柱层析分离得到中间体2-氯-4-((1r,3r)-3-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(536mg,1.25mmol)。收率:66%。LC/MS(ESI+)calcd for C22H21Cl2N3O2(M+H+)m/z,430.1;found,430.1.
SM-A-3:4-((1r,3r)-3-(5-溴-1-异吲哚啉-2-基)-2,2,4,4-四甲基环丁醚)-2-氯苯腈的合成
化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁基醚)-2-氯苯腈(1.0g,3.59mmol)和K2CO3(743.0mg,5.38mmol)加入到20mL DMF中,将4-溴-2-(溴甲基)苯甲酸甲脂(1.10g,3.59mmol)分批加入到反应液中。室温搅拌1小时,TLC监测反应完全,生成一个新点。加入乙酸乙酯和饱和食盐水萃取,有机层饱和食盐水洗涤2次,无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到油状液体1.2g,加入甲苯10mL,三乙胺1mL,加热回流,搅拌16h。溶剂旋干,加入少量石油醚和乙酸乙酯(4:1),固体抽滤,干燥,得到白色固体4-((1r,3r)-3-(5-溴-1-异吲哚啉-2-基)-2,2,4,4-四甲基环丁基醚)-2-氯苯腈(900.0mg,1.9mmol)。收率:52.9%。
SM-A-4:4-(((1r,4r)-4-(5-溴-1-氧异吲哚啉-2-基)环己基)氧基)-2-氯苯甲腈
第一步:合成4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈
化合物反-对氨基环己醇盐酸盐(26.8g,177mmol)溶于DMF 0.8L,冰浴,N2保护,分批加入NaH(22.3g,531mmol),0℃反应1h。化合物2-氯-4-氟-苯甲腈溶于200ml DMF,缓慢滴加到反应中,0℃保温0.5h,回复至常温,搅拌3h,点板反应完全。加入EA和水萃取,EA再用饱和食盐水洗涤2次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体产物4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(27.7g)。
第二步:
化合物4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(1g,4mmol)溶于20mLDMF,加入K2CO3(0.82g,6mmol)。分批加入化合物4-溴-2-(溴甲基l)苯甲酸甲酯(1.2g,4mmol)。室温搅拌过夜。加水,析出白色固体,抽滤,固体干燥,得到化合物SM-A-3(1.3g,2.9mmol)。
SM-A-5:2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
将化合物4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(2.37g,9.45mmol)、碳酸钾(1.96g,14.18mmol)和40ml DMF依次加入,氮气保护,搅拌均匀后滴入2-(溴甲基)-6-氯烟酸甲酯(2.5g,9.45mmol)的DMF溶液,室温搅拌2h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到中间体。中间体加入DIEA(4.73mg,36.61mmol)和20ml 1,4二氧六环,100℃反应2d,TLC确定反应终点硅胶柱层析分离得到中间体2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(1.85g,4.60mmol)。收率:49%。
LC/MS(ESI+)calcd for C20H17Cl2N3O2(M+H+)m/z,402.1;found,402.1.
SM-A-6:2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲硫基)-5-氧-5,7-二氢-6H-吡咯[3,4-d]嘧啶-6-基)环丁氧基)苯甲腈
1.4-(溴甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯
将4-甲基-2-(甲硫基)嘧啶-5-羧酸乙酯(10.6g,50.0mmol)溶入醋酸(30mL)中,在冰浴中向反应液中滴加溴素(9.6g,60.0mmol)。反应液加热到60℃反应3小时。反应液倒入冰水(100mL)中,然后用乙酸乙酯(50mL×3)萃取。合并有机相用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=50:1至30:1)纯化得到黄色油状物4-(溴甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯(8.3g,28.6mmol),收率:57%。MS:calcd.forC9H11BrN2O2S[M+H]+:291.0;found:291.0.
2.4-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁胺基)甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯
将4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈三氟乙酸盐(11.4mmol,粗品来源于先前的路线)溶入乙腈(120mL)中,然后加入N,N-二异丙基乙胺(7.35g,11.4mmol)和4-(溴甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯(4.3g,11.4mmol)。反应液室温搅拌过夜。反应液用水(100mL)稀释,乙酸乙酯(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=50:1至10:1)纯化得到类白色固体4-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁胺基)甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯(3.4g,7.0mmol),收率:61%。MS:calcd.for C24H29ClN4O3S[M+H]+:489.2;found:489.2.
3.2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲硫基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(SM-A-6)
将4-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁胺基)甲基)-2-(甲硫基)嘧啶-5-羧酸乙酯(2.0g,4.1mmol)溶入甲苯(100mL)中,然后置换氮气,并在冰浴中滴加三甲基铝的正己烷溶液(2.0M)(4.1mL,8.2mmol)。反应液加热到110℃反应20小时。反应液用水(100mL)稀释,乙酸乙酯(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=50:1至30:1)纯化得到黄色固体2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲硫基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(0.8g,1.8mmol),收率:49%。MS:calcd.for C22H23ClN4O2S[M+H]+:443.1;found:443.1.
其他内酰胺类中间体用与上述路线类似的方法合成得到。
以下酰胺类中间体化合物物用文献记载的方法(US20180099940,US20170327469)或与其类似的方法合成得到。
以下SM-E中间体化合物用文献记载的方法(US20180099940,US20170327469)或与其类似的方法合成得到。
中间体SM-L-1:4-乙炔基-1,4'-二哌啶-1'-羧酸叔丁酯
1.化合物4-(对甲基苯磺酰氧基)哌啶-1-羧酸叔丁酯
将4-羟基哌啶-1-羧酸叔丁酯(10.0g,49.7mmol),溶入二氯甲烷中,然后加入对甲基苯磺酰氯(10.8g,56.6mmol),三乙胺(7.5g,74.5mmol)和4-二甲氨基吡啶(183mg,1.5mmol)。反应液室温搅拌过夜。反应液用水(100mL)稀释,二氯甲烷(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至3:1)纯化得到白色固体4-(对甲基苯磺酰氧基)哌啶-1-羧酸叔丁酯(13.2g,37.2mmol),收率:83%。MS:calcd.for C17H25NO5S[M+H]+:356.1;found:356.1.
2.化合物4-乙炔基-1,4'-二哌啶-1'-羧酸叔丁酯(SM-L-1)的合成
将4-(对甲基苯磺酰氧基)哌啶-1-羧酸叔丁酯(1.2g,3.4mmol)溶入乙腈中,然后加入4-乙炔哌啶盐酸盐(490mg,3.4mmol),碳酸钾(1.0g,7.5mmol)和碘化钾(113mg,0.68mmol)。反应液加热回流36小时。反应液用水(50mL)稀释,乙酸乙酯(20mL×3)萃取。有机相合并用饱和食盐水(20mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至纯乙酸乙酯)纯化得到白色固体4-乙炔基-1,4'-二哌啶-1'-羧酸叔丁酯(0.6g,2.1mmol),收率:60%。MS:calcd.for C17H28N2O2[M+H]+:293.1;found:293.1.
用与合成SM-L-1类似的方法制备得到SM-L-3:
中间体SM-L-2:3-(4-乙炔基哌啶-1-基)氮杂环丁烷-1-羧酸叔丁酯
将4-乙炔哌啶盐酸盐(2.0g,13.8mmol)和3-羧酸叔丁酯3-氮杂环丁酮(2.35g,13.8mmol)溶入1,2-二氯乙烷(40mL),随后加入乙酸(1mL),最后加入三乙酰氧基硼氢化钠(5.85g,27.6mmol)。反应液室温搅拌过夜。反应液用水(50mL)稀释,二氯甲烷(30mL×3)萃取。有机相合并用饱和食盐水(30mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至纯乙酸乙酯)纯化得到无色油状物3-(4-乙炔基哌啶-1-基)氮杂环丁烷-1-羧酸叔丁酯(1.0g,3.8mmol),收率:27%。MS:calcd.for C15H24N2O2[M+H]+:265.2;found:265.2.
中间体SM-L-4,SM-L-8,SM-L-9和SM-L-10的合成与SM-L-2类似。
中间体SM-L-11:4-(2-甲基-3-炔-2-基)哌嗪-1-羧酸叔丁酯
将哌嗪-1-羧酸叔丁酯(3.3g,17.7mmol),三乙胺(3.57g,35.4mmol)和3-氯-3-甲基丁炔(1.8g,17.7mmol)溶入四氢呋喃(30mL),然后加入氯化亚铜(179mg,1.8mmol)。反应液室温搅拌10分钟。反应液用水(100mL)稀释,乙酸乙酯(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至2:1)纯化得到无色油状物4-(2-甲基-3-炔-2-基)哌嗪-1-羧酸叔丁酯(3.0g,11.9mmol),收率:67%。MS:calcd.for C14H24N2O2[M+H]+:253.2;found:253.2.
中间体SM-L-13:4-(2-甲基-3-炔-2-基氨基)哌啶-1-羧酸叔丁酯(
将4-氨基哌啶-1-羧酸叔丁酯(2.6g,13.0mmol),三乙胺(1.51g,15.0mmol)和溴化亚铜(144mg,1.0mmol)加入乙腈(30mL)中,然后向反应液中滴加3-氯-3-甲基丁炔(1.0g,10.0mmol)的乙腈(10mL)溶液。反应液室温搅拌1小时。反应液用水(100mL)稀释,乙酸乙酯(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至2:1)纯化得到无色油状物4-(2-甲基-3-炔-2-基氨基)哌啶-1-羧酸叔丁酯(0.6g,2.3mmol),收率:22%。MS:calcd.for C15H26N2O2[M+H]+:267.2;found:267.2.
中间体SM-L-14:4-(4-乙炔基苯基)哌嗪-1-羧酸叔丁酯
1.化合物4-(4-甲酰苯基)哌嗪-1-羧酸叔丁酯
将4-氟苯甲醛(5.0g,40.3mmol)和哌嗪-1-羧酸叔丁酯(7.5g,40.3mmol)溶入N,N-二甲基甲酰胺(100mL)中,然后加入碳酸钾(11.2g,80.6mmol)。反应液在110℃搅拌2天。反应液用水(150mL)稀释,乙酸乙酯(100mL×3)萃取。有机相合并用饱和食盐水(100mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=20:1至5:1)纯化得到白色固体4-(4-甲酰苯基)哌嗪-1-羧酸叔丁酯(7.1g,24.5mmol),收率:61%。
MS:calcd.for C16H23O2N3[M+H]+:290.2;found:290.0.
2.化合物4-(4-乙炔基苯基)哌嗪-1-羧酸叔丁酯
将4-(4-甲酰苯基)哌嗪-1-羧酸叔丁酯(B-14-1)(2.9g,10.0mmol)和1-重氮-2-氧丙基膦酸二甲酯(3.8g,20.0mmol)溶入甲醇(50mL),在冰浴中加入碳酸钾(5.5g,40mmol)。反应液缓慢升温至室温过夜。反应液旋干,硅胶柱层析(石油醚:乙酸乙酯=50:1至15:1)纯化得到白色固体4-(4-乙炔基苯基)哌嗪-1-羧酸叔丁酯(1.5g,5.2mmol),收率:54%。MS:calcd.for C17H23O2N2[M+H]+:287.2;found:287.2.
中间体SM-L-18:4-((1s,3s)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18Q)和4-((1r,3r)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18H)
1.化合物4-(3-(甲氧羰基)环丁基)哌嗪-1-羧酸叔丁酯
将3-氧环丁烷羧酸甲酯(5.12g,40.0mmol)和哌嗪-1-羧酸叔丁酯(8.9g,48.0mmol)溶入1,2-二氯乙烷(100mL,随后加入乙酸(4.8g,80.0mmol),最后加入三乙酰氧基硼氢化钠(21.2g,100mmol)。反应液室温搅拌过夜。反应液用水(100mL)稀释,二氯甲烷(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=10:1至纯乙酸乙酯)纯化得到白色固体4-(3-(甲氧羰基)环丁基)哌嗪-1-羧酸叔丁酯(6.0g,20.8mmol),收率:52%。MS:calcd.for C15H26N2O4[M+H]+:299.2;found:299.4.
2.化合物4-(3-(羟甲基)环丁基)哌嗪-1-羧酸叔丁酯
将4-(3-(甲氧羰基)环丁基)哌嗪-1-羧酸叔丁酯(6.0g,20.8mmol)溶入四氢呋喃(120mL),在冰浴中分批加入四氢锂铝(4.8g,80.0mmol)。反应液在冰浴中继续搅拌1小时。反应液用四氢呋喃稀释(100mL),保持在冰浴中,然后滴加水(4.8mL)搅拌10分钟,随后加入15%的氢氧化钠溶液(4.8mL)继续搅拌10分钟,再加入水(14.4mL)继续搅拌20分钟,最后加入无水硫酸镁搅拌20分钟。反应混合物过滤,滤液旋干得到无色油状物4-(3-(羟甲基)环丁基)哌嗪-1-羧酸叔丁酯(4.4g,14.8mmol),收率:78%。MS:calcd.for C14H26N2O3[M+H]+:271.2;found:271.4.
3.化合物4-(3-甲酰基环丁基)哌嗪-1-羧酸叔丁酯
将4-(3-(羟甲基)环丁基)哌嗪-1-羧酸叔丁酯(4.0g,14.8mmol)溶入二氯甲烷(80mL)中,然后加入1,1',1'-(3-氧代-1λ5-1,2-苯碘代-1(3H)-叶立酮)乙酰乙酸酯(9.4g,22.2mmol)。反应液室温搅拌24小时。然后过滤,滤液旋干得到4-(3-甲酰基环丁基)哌嗪-1-羧酸叔丁酯粗品(5.3g),直接用于下一步反应。MS:calcd.for C14H24N2O3[M+H]+:269.2;found:269.4.
4.化合物4-((1s,3s)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18Q)和4-((1r,3r)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18H)的合成
将4-(3-甲酰基环丁基)哌嗪-1-羧酸叔丁酯(14.8mmol,粗品来源于上一步)和1-重氮-2-氧丙基膦酸二甲酯(5.68g,29.6mmol)溶入甲醇(100mL),在冰浴中加入碳酸钾(10.2g,74.0mmol)。反应液缓慢升温至室温过夜。反应液用水(200mL)稀释,乙酸乙酯(80mL×3)萃取。有机相合并用饱和食盐水(80mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=20:1至12:1)纯化得到类白色固体4-((1s,3s)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18Q)(0.84g,3.2mmol),两步收率:16%。MS:calcd.for C15H24N2O2[M+H]+:265.2;found:265.4。同时得到类白色固体4-((1r,3r)-3-乙炔基环丁基)哌嗪-1-羧酸叔丁酯(SM-L-18H)(0.36g,1.4mmol),两步收率:7%。MS:calcd.for C15H24N2O2[M+H]+:265.2;found:265.4.
中间体SM-L-5的合成
1.化合物4-(4-(甲氧羰基)环己基)哌嗪-1-羧酸叔丁酯的合成
将4-氧代环己基甲酸甲酯(10.0g,64.0mmol)和哌嗪-1-羧酸叔丁酯(13.0g,70.0mmol)溶入1,2-二氯乙烷(200mL,随后加入乙酸(7.6g,128.0mmol),最后加入三乙酰氧基硼氢化钠(34.0g,160mmol)。反应液室温搅拌过夜。反应液用水(200mL)稀释,二氯甲烷(100mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=1:1)纯化得到白色固体4-(4-(甲氧羰基)环己基)哌嗪-1-羧酸叔丁酯(9.6g,29.4mmol),收率:46%。MS:calcd.for C17H30N2O4[M+H]+:327.2;found:327.4.
2.化合物4-((1r,4r)-4-(羟甲基)环己基)哌嗪-1-羧酸叔丁酯的合成
将4-(4-(甲氧羰基)环己基)哌嗪-1-羧酸叔丁酯(9.6g,29.4mmol)溶入四氢呋喃(200mL),在冰浴中分批加入四氢锂铝(2.23g,58.8mmol)。反应液在冰浴中继续搅拌1小时。反应液用四氢呋喃稀释(100mL),保持在冰浴中,然后滴加水(2.3mL)搅拌10分钟,随后加入15%的氢氧化钠溶液(2.3mL)继续搅拌10分钟,再加入水(6.9mL)继续搅拌20分钟,最后加入无水硫酸镁搅拌20分钟。反应混合物过滤,滤液旋干,硅胶柱层析(石油醚:乙酸乙酯=1:1至乙酸乙酯)纯化得到浅黄色油状物4-((1r,4r)-4-(羟甲基)环己基)哌嗪-1-羧酸叔丁酯(1.2g,4.0mmol),收率:14%。构型参考专利WO2012145361确定。MS:calcd.for C14H26N2O3[M+H]+:299.2;found:299.4.1H NMR(400MHz,CDCl3):δ3.46–3.33(m,6H),2.51–2.38(m,4H),2.28–2.17(m,1H),1.91–1.84(m,4H),1.62–1.48(m,1H),1.42(s,9H),1.27–1.13(m,3H),1.02–0.88(m,2H).
同时得到浅黄色油状物4-((1s,4s)-4-(羟甲基)环己基)哌嗪-1-羧酸叔丁酯(3.0g,10.0mmol),收率:34%。MS:calcd.for C14H26N2O3[M+H]+:299.2;found:299.4.
3.化合物4-((1r,4r)-4-(甲酰基)环己基)哌嗪-1-羧酸叔丁酯的合成
将4-((1r,4r)-4-(羟甲基)环己基)哌嗪-1-羧酸叔丁酯(1.2g,4.0mmol)溶入二氯甲烷(50mL)中,然后加入1,1',1'-(3-氧代-1λ5-1,2-苯碘代-1(3H)-叶立酮)乙酰乙酸酯(2.0g,4.8mmol)。反应液室温搅拌24小时。然后过滤,滤液旋干得到4-((1r,4r)-4-(甲酰基)环己基)哌嗪-1-羧酸叔丁酯粗品(1.4g),直接用于下一步反应。MS:calcd.forC16H28N2O3[M+H]+:297.2;found:297.4.
4.化合物4-((1r,3r)-4-乙炔基环己基)哌嗪-1-羧酸叔丁酯的合成
将4-((1r,4r)-4-(甲酰基)环己基)哌嗪-1-羧酸叔丁酯(4.0mmol,粗品来源于上一步方法一)和1-重氮-2-氧丙基膦酸二甲酯(7.2g,6.0mmol)溶入甲醇(50mL),在冰浴中加入碳酸钾(1.1g,8.0mmol)。反应液缓慢升温至室温过夜。反应液用水(50mL)稀释,乙酸乙酯(50mL×3)萃取。有机相合并用饱和食盐水(50mL×3)洗涤,硫酸钠干燥,硅胶柱层析(石油醚:乙酸乙酯=3:1至2:1)纯化得到浅黄色固体4-((1r,3r)-3-乙炔基环己基)哌嗪-1-羧酸叔丁酯(0.67g,2.3mmol),两步收率:57%。MS:calcd.for C17H28N2O2[M+H]+:293.2;found:293.2.1H NMR(400MHz,CDCl3):δ3.46–3.36(m,4H),2.53–2.44(m,4H),2.35–2.23(m,1H),2.22–2.11(m,1H),2.09–2.00(m,3H),1.93–1.83(m,2H),1.45(s,9H),1.43–1.34(m,2H),1.28–1.20(m,2H).
中间体SM-L-6,SM-L-7,SM-L-15,SM-L-16,SM-L-17,SM-L-18,SM-L-19,SM-L-20,SM-L-21,SM-L-22,SM-L-23和SM-L-24的合成与SM-L-18类似。
以下为本发明提供的具体化合物的制备实例:
1:(3R,5S)-1-((S)-2-(2-((5-((4-(3-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基))苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯
1.合成2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)乙基)戊基)氧基)氨基叔丁酯
将1-(4-氨基苯基)-3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲(100.0mg,0.24mmol),2-((5-氧代戊基)氧基)乙酸叔丁酯(52.4mg,0.24mmol)溶于5ml DCM,降温至0℃,加入一滴乙酸,然后分批次加入氰基硼氢化钠(45.6mg,0.73mmol),恢复至室温搅拌过夜。加入水,DCM萃取,干燥旋干,Pre-TLC纯化得到化合物2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)乙基)戊基)氧基)氨基叔丁酯73.4mg。收率:54.6%。LC/MS(ESI+)Calcd for C33H45ClN4O5(M-56+H+)m/z,612.3;Found,557.3。
2.2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧)乙酸
将2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)乙基)戊基)氧基)氨基叔丁酯(70.0mg,0.09mmol)溶于2ml DCM,加入5ml TFA,室温搅拌1h,TLC检测原料反应完全。旋干体系,加入DCM溶解,用饱和碳酸氢钠水溶液洗至PH约为6。干燥有机相,过滤浓缩。得到2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧)乙酸21.3mg。收率:32.6%。
3.合成(3R,5S)-1-((S)-2-(2-((5-((4-(3-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基))苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯
将2-((5-((4-(3-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧)乙酸(20.0mg,0.09mmol)溶于2ml DMF,降温至0℃左右,加入0.5mlDIEA,以及HATU(29.1mg,0.13mmol),搅拌5min。然后加入(3R,5S)-1-((S)-2-氨基-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-3-乙酸酯(26.0mg,0.10mmol),恢复至室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(3R,5S)-1-((S)-2-(2-((5-((4-(3-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)脲基)苯基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基))苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯8.2
mg。收率:37.2%。LC/MS(ESI+)Calcd for C54H69ClN8O8S(M+H+)m/z,1024.5;Found,1024.4。1H NMR(400MHz,DMSO-d6)δ9.21(m,1H),9.11(m,1H),8.87-8.82(d,J=7.6Hz,1H),8.13(m,1H),7.90(d,J=8.8Hz,1H),7.38(dt,J=8.7,7.5Hz,6H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),6.85(d,J=8.3Hz,1H),6.77(s,1H),5.87(s,1H),5.20(s,1H),4.93–4.88(m,1H),4.60(s,2H),4.53(s,1H),4.45(dd,J=14.4,8.7Hz,2H),4.29(s,1H),3.90(dd,J=16.2,6.4Hz,3H),3.77(dd,J=11.8,3.9Hz,1H),3.49(t,J=6.4Hz,2H),3.05(s,2H),2.45(s,3H),2.29–2.23(m,1H),2.09(s,1H),1.99(s,3H),1.98(d,J=3.1Hz,1H),1.62–1.56(m,4H),1.45(d,J=4.9Hz,3H),1.39(s,6H),1.31(s,1H),1.21(s,6H),0.93(s,9H).
2:(3R,5S)-1-((S)-2-(2-(2-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯
1.化合物2-(2-(4-(2–((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉)哌嗪-1-基)乙氧基)-5-乙酸叔丁酯的合成
将4-((1r,3r)-3-(5-溴-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(100.0mg,0.21mmol),2-(2-(哌嗪-1-基)乙氧基)乙酸叔丁酯(154.5mg,0.63mmol),BINAP(39.0mg,0.633mmol),Pb2(dba)3(19.1mg,0.02mmol),Cs2CO3(137.5mg,0.42mmol)溶于10ml甲苯,氮气置换后升温至100℃,反应过夜。垫硅藻土过滤,浓缩滤液,加入水,二氯甲烷萃取。干燥有机相,过滤浓缩,Pre-TLC纯化得到化合物2-(2-(4-(2–((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉)哌嗪-1-基)乙氧基)-5-乙酸叔丁酯94.7mg。收率:49.3%。LC/MS(ESI+)Calcd for C35H45ClN4O5(M-56+H+)m/z,636.3;Found,581.2。
2.化合物2-(2-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁烷)-1-二氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酸的合成
将2-(2-(4-(2–((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉)哌嗪-1-基)乙氧基)-5-乙酸叔丁酯(90.0mg,0.19mmol)溶于1ml二氯甲烷,然后加入5ml三氟乙酸室温搅拌30min。TLC监测反应完毕直接旋干,加入二氯甲烷溶解,用饱和碳酸氢钠调PH=6,分液,用无水硫酸钠干燥有机相,过滤旋干得到化合物2-(2-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁烷)-1-二氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酸63.3mg。收率:89.3%。
3.(3R,5S)-1-((S)-2-(2-(2-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯的合成
将2-(2-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁烷)-1-二氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酸(30.0mg,0.09mmol)溶于2ml DMF,降温至0℃左右,加入0.5ml DIEA,以及HATU(29.1mg,0.13mmol),搅拌5min。然后加入(3R,5S)-1-((S)-2-氨基-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-3-乙酸酯(26.0mg,0.10mmol),恢复至室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(3R,5S)-1-((S)-2-(2-(2-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯11.6mg。收率:33.5%。LC/MS(ESI+)Calcdfor C56H69ClN8O8S(M+H+)m/z,1048.5;Found,1048.5。1H NMR(400MHz,DMSO-d6)δ9.02(m,1H),8.98–8.93(m,1H),8.47(d,=7.6Hz,1H),7.97(d,J=8.8Hz,1H),7.38(dt,J=8.7,7.5Hz,6H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),6.85(d,J=8.3Hz,1H),6.77(s,1H),5.87(s,1H),5.20(s,1H),4.93–4.88(m,1H),4.60(s,2H),4.53(s,1H),4.45(dd,J=14.4,8.7Hz,2H),4.29(s,1H),4.22–4.05(m,4H),3.90(dd,J=16.2,6.4Hz,3H),3.77(dd,J=11.8,3.9Hz,1H),3.49(t,J=6.4Hz,2H),3.05(s,2H),2.29–2.23(m,1H),2.09(s,1H),1.99(s,3H),1.98(d,J=3.1Hz,1H),1.45(d,J=4.9Hz,3H),1.37(s,6H),1.35(s,1H),1.14(s,6H),0.95(s,9H).
3:化合物(2S,4R)-1-((S)-2-(2-(2-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将2-(2-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁烷)-1-二氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酸(30.0mg,0.09mmol)溶于2ml DMF,降温至0℃左右,加入0.5ml DIEA,以及HATU(29.1mg,0.13mmol),搅拌5min。然后加入(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(24.0mg,0.10mmol),恢复至室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(3R,5S)-1-((S)-2-(2-(2-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)乙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯8.2mg。收率:31.7%。LC/MS(ESI+)Calcd for C54H67ClN8O7S(M+H+)m/z,1007.5;Found,1007.5。1H NMR(400MHz,DMSO-d6)δ9.14(m,1H),9.02(m,1H),8.47(d,J=7.6Hz,1H),7.97(d,J=8.8Hz,1H),7.38(dt,J=8.7,7.5Hz,6H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),6.85(d,J=8.3Hz,1H),6.77(s,1H),5.87(s,1H),5.20(s,1H),4.93–4.88(m,1H),4.60(s,2H),4.53(s,1H),4.45(dd,J=14.4,8.7Hz,2H),4.29(s,1H),4.22–4.05(m,4H),3.90(dd,J=16.2,6.4Hz,3H),3.77(dd,J=11.8,3.9Hz,1H),3.49(t,J=6.4Hz,2H),3.05(s,2H),2.45(s,3H),2.29–2.23(m,1H),2.09(s,1H),1.99(s,3H),1.98(d,J=3.1Hz,1H),1.45(d,J=4.9Hz,3H),1.37(s,6H),1.35(s,1H),1.14(s,6H),0.95(s,9H).
4:(3R,5R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯
1.化合物2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸叔丁酯的合成
将4-((1r,3r)-3-)-6-溴-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(85.0mg,0.18mmol),PbdppfCl2(26.5mg,0.04mmol),CuI(17.5mg,0.09mmol),2-(己-5-炔-1-基氧基)乙酸叔丁酯(38.5mg,0.18mmol),依次加入反应瓶。氮气置换,注射器加入甲苯1.2ml,三乙胺0.4ml,再次氮气置换。110℃反应过夜,TLC检测反应完全。将体系冷却至室温,垫硅藻土过滤,滤液浓缩。用二氯甲烷溶解,分别0.5M HCl,饱和食盐水洗有机相,干燥过滤,真空浓缩。Pre-TLC纯化,得到2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸叔丁酯97.3mg。收率:78.8%。LC/MS(ESI+)Calcd for C35H41ClN2O5(M-56+H+)m/z,604.3;Found,549.2。
2.2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸的合成
将2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸叔丁酯(95.0mg,0.15mmol)溶于1ml DCM,加入5ml TFA,室温反应1h,TLC检测原料反应完全。旋干体系,加入DCM溶解,用饱和碳酸氢钠水溶液洗至PH约为6。干燥有机相,过滤浓缩。得到2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸81.2mg。收率:92.6%。
3.(3R,5R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(4)
将2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)己-5-炔-1-基)氧基)乙酸(80.0mg,0.12mmol)溶于2ml DMF,加入DIEA(98.5mg,0.76mmol),降温至0℃左右,加入HATU(96.0mg,0.18mmol),搅拌10min后加入(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(130.1mg,0.22mmol),恢复至室温反应2h。TLC检测反应完毕。将体系缓慢加入5ml H2O中,用乙酸乙酯萃取,有机相再用水洗涤5次,然后分别用0.5MHCl,饱和食盐水洗。干燥过滤,真空浓缩。Pre-TLC纯化得到(3R,5R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯38.0mg。收率:53.8%。1H NMR(400MHz,CDCl3)δ8.69(s,1H),7.88(s,1H),7.59-7.54(m,2H),7.38(dd,J=16.0,8.3Hz,6H),7.19(d,J=9.2Hz,1H),6.99(d,J=2.4Hz,1H),6.83(dd,J=8.7,2.4Hz,1H),5.35(s,1H),5.10-5.05(m,1H),4.75(dd,J=8.3,6.5Hz,1H),4.65(s,2H),4.59(d,J=9.3Hz,1H),4.39(s,1H),4.31(s,1H),4.05(d,J=13.0Hz,1H),3.96(d,J=6.2Hz,2H),3.83(dd,J=11.6,4.9Hz,1H),3.59(t,J=6.2Hz,2H),2.74-2.68(m,1H),2.53(s,3H),2.50(s,1H),2.16-2.10(m,1H),2.03(d,J=8.6Hz,4H),1.86-1.80(m,2H),1.73(dd,J=11.5,4.6Hz,2H),1.47(d,J=6.9Hz,3H),1.44(t,J=3.1Hz,6H),1.25(s,6H),1.05(s,9H)。LC/MS(ESI+)Calcd forC56H65ClN6O8S(M/2+H+)m/z,1016.5;Found,509.3。
5:(2R,4R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(5)
将(3R,5R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(5.0mg,4.54μmol)溶于1ml甲醇,缓慢加入LiOH.H2O(0.9mg,0.02mmol)与0.5ml H2O的混合体系。大约30min TLC监测反应完毕。加入水,DCM萃取。Pre-TLC纯化得到化合物(2R,4R)-1-((S)-2-(2-((6-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺3.0mg。收率:84.3%。1H NMR(400MHz,CDCl3)δ8.71(s,1H),7.86(s,1H),7.57(dd,J=7.6,4.6,2H),7.53-7.46(m,1H),7.38(q,J=8.5,5H),7.23(d,J=9.0,1H),6.99(d,J=2.3,1H),6.83(dd,J=8.7,2.3,1H),5.11-5.03(m,1H),4.78(t,J=7.9,1H),4.65(s,2H),4.54(d,J=8.4,2H),4.40(s,1H),4.31(s,1H),4.15(d,J=11.3,1H),3.95(t,J=11.5,2H),3.60(t,J=6.1,3H),2.54(s,4H),2.50(t,J=6.7,2H),2.13-1.98(m,2H),1.82(d,J=6.6,2H),1.78-1.73(m,2H),1.47(d,J=6.9,3H),1.44(s,6H),1.25(s,6H),1.07(s,9H)。LC/MS(ESI+)Calcd for C54H63ClN6O7S(M+H+)m/z,975.4;Found,975.5。
6:(3R,5S)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5)-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(6)
1.2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸叔丁酯的合成
将4-((1r,3r)-3-(6-氨基-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(150.0mg,0.37mmol),2-((5-氧代戊基)氧基)乙酸叔丁酯(79.1mg,0.35mmol)溶于5ml DCM,降温至0℃,加入一滴乙酸,然后分批次加入氰基硼氢化钠(58.6mg,0.92mmol),恢复至室温搅拌过夜。加入水,DCM萃取,干燥旋干,Pre-TLC纯化得到化合物2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸叔丁酯74.4mg。收率:67.5%。LC/MS(ESI+)Calcd forC34H44ClN3O5(M-56+H+)m/z,609.3;Found,554.2。
2.2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸的合成
将2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸叔丁酯(74.0mg,0.12mmol)溶于1ml DCM,加入5mlTFA,室温搅拌1h。TLC检测原料反应完全。旋干体系,加入DCM溶解,用饱和碳酸氢钠水溶液洗至PH约为6。干燥有机相,过滤浓缩。得到2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸70.3mg。收率:96.3%。
3.(3R,5S)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5)-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯的合成
将2-((2-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚-5-基)氨基)戊基)氧基)乙酸(70.0mg,0.13mmol)溶于4ml DMF,加入DIEA(81.5mg,0.63mmol),降温至0℃左右,加入HATU(72.0mg,0.19mmol),搅拌10min后加入(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(92.1mg,0.19mmol),恢复至室温反应2h。TLC检测反应完毕。将体系缓慢加入5ml H2O中,用乙酸乙酯萃取,有机相再用水洗涤5次,然后分别用0.5MHCl,饱和食盐水洗。干燥过滤,真空浓缩。Pre-TLC纯化得到(3R,5S)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5)-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯31.3mg。收率:38.8%。1H NMR(400MHz,DMSO-d6)δ8.98(m,1H),8.47(d,J=7.6Hz,1H),7.90(d,J=8.8Hz,1H),7.38(dt,J=8.7,7.5Hz,6H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),6.85(d,J=8.3Hz,1H),6.77(s,1H),5.87(s,1H),5.20(s,1H),4.93-4.88(m,1H),4.60(s,2H),4.53(s,1H),4.45(dd,J=14.4,8.7Hz,2H),4.29(s,1H),3.90(dd,J=16.2,6.4Hz,3H),3.77(dd,J=11.8,3.9Hz,1H),3.49(t,J=6.4Hz,2H),3.05(s,2H),2.45(s,3H),2.29-2.23(m,1H),2.09(s,1H),1.99(s,3H),1.98(d,J=3.1Hz,1H),1.62-1.56(m,4H),1.45(d,J=4.9Hz,3H),1.37(s,6H),1.35(s,1H),1.14(s,6H),0.95(s,9H)。LC/MS(ESI+)Calcd for C55H68ClN7O8S(M+H+)m/z,1021.5;Found,1022.5。
7:(2S,4R)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(7)
将(3R,5S)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5)-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(5.0mg,4.30μmol)溶于1ml甲醇,缓慢加入LiOH.H2O(0.9mg,0.02mmol)与0.5ml H2O的混合体系。大约30minTLC监测反应完毕。加入水,DCM萃取。Prep-TLC纯化得到化合物(2S,4R)-1-((S)-2-(2-((5-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代异吲哚啉-5-基)氨基)戊基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺3.3mg。收率:87.4%。1H NMR(400MHz,DMSO-d6)δ8.97(m,1H),8.46(d,J=7.6Hz,1H),7.89(d,J=8.8Hz,1H),7.38(dt,J=8.7,7.5Hz,6H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),6.85(d,J=8.3Hz,1H),6.77(s,1H),5.87(s,1H),5.20(s,1H),4.93-4.88(m,1H),4.60(s,2H),4.53(s,1H),4.45(dd,J=14.4,8.7Hz,2H),4.29(s,1H),3.90(dd,J=16.2,6.4Hz,3H),3.77(dd,J=11.8,3.9Hz,1H),3.49(t,J=6.4Hz,2H),3.05(s,2H),2.45(s,3H),2.28-2.22(m,1H),2.08(s,1H),1.98(d,J=3.1Hz,1H),1.62-1.54(m,4H),1.43(d,J=4.9Hz,3H),1.36(s,6H),1.35(s,1H),1.13(s,6H),0.95(s,9H)。LC/MS(ESI+)Calcd for C53H66ClN7O7S(M+H+)m/z,979.4;Found,980.3。
8:(3R,5R)-1-((S)-2-(2-((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4-,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((1-(4-(4-甲基噻唑-5-基)苯基)环丙基)氨基甲酰基)吡咯烷-3-基乙酸酯(8)
1.N-((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘苯甲酰胺的合成
将对碘苯甲酸(103.0mg,0.42mmol)溶于2ml DCM,降温至0℃左右,依次加入DIEA(266.6mg,2.08mmol),HATU(237.2mg,0.62mmol)。低温下搅拌10min,加入4–((1R,3R)-3-氨基-2,2,4,4-四甲基环丁基)-2-氯苄腈三氟乙酸盐(200.0mg,0.46mmol),撤去冰浴,室温反应2h,TLC检测反应完毕。反应体系用0.5M HCl水溶液洗,饱和食盐水洗,干燥,真空浓缩至不滴,通过柱纯化得到N-((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘苯甲酰胺221.0mg。收率:73.7%。LC/MS(ESI+)Calcd for C22H22BrIN2O2(M+H+)m/z,554.0;Found,555.1。
2.2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸叔丁酯的合成
将N-((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘苯甲酰胺(100.0mg,0.18mmol),PbdppfCl2(26.5mg,0.04mmol),CuI(17.5mg,0.09mmol),2-(己-5-炔-1-基氧基)乙酸叔丁酯(38.5mg,0.18mmol),依次加入反应瓶。氮气置换,注射器加入甲苯1.2ml,三乙胺0.4ml,再次氮气置换。90℃反应3h,TLC检测反应完全。将体系冷却至室温,垫硅藻土过滤,滤液浓缩。用二氯甲烷溶解,分别0.5M HCl,饱和食盐水洗有机相,干燥过滤,真空浓缩。Pre-TLC纯化,得到2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸叔丁酯121.3mg。收率:91.6%。LC/MS(ESI+)Calcd for C34H41BrN2O5(M-56+H+)m/z,638.2;Found,583.2。
3.2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸的合成
将2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸叔丁酯(120.0mg,0.19mmol)溶于1ml DCM,加入5mlTFA,室温反应1h,TLC检测原料反应完全。旋干体系,加入DCM溶解,用饱和碳酸氢钠水溶液洗至PH约为6。干燥有机相,过滤浓缩。得到2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸95.2mg。收率:92.4%。
4.(3R,5R)-1-((S)-2-(2-((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4-,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((1-(4-(4-甲基噻唑-5-基)苯基)环丙基)氨基甲酰基)吡咯烷-3-基乙酸酯的合成
将2-(6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基-苯基)己-5-炔-1-基氧基)乙酸(95.0mg,0.16mmol)溶于2ml DMF,加入DIEA(113.5mg,0.89mmol),降温至0℃左右,加入HATU(101.0mg,0.24mmol),搅拌10min后加入(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-环丙酰胺(130.1mg,0.22mmol),恢复至室温反应2h。TLC检测反应完毕。将体系缓慢加入5ml H2O中,用乙酸乙酯萃取,有机相再用水洗涤5次,然后分别用0.5MHCl,饱和食盐水洗。干燥过滤,真空浓缩。Pre-TLC纯化得到(3R,5R)-1-((S)-2-(2-((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((1-(4-(4-甲基噻唑-5-基)苯基)环丙基)氨基甲酰基)吡咯烷-3-基乙酸酯40.0mg。收率:46.7%。1H NMR(400MHz,CDCl3)δ8.81(s,1H),7.70(d,J=8.1Hz,2H),7.56(d,J=8.7Hz,1H),7.45(d,J=8.4Hz,3H),7.33(d,J=3.6Hz,4H),7.15(d,J=2.3Hz,2H),6.85(dd,J=8.7,2.3Hz,1H),6.27(d,J=8.3Hz,1H),5.37(s,1H),4.64(t,J=7.4Hz,1H),4.54(d,J=9.3Hz,1H),4.15(d,J=8.2Hz,1H),4.06(s,1H),4.01-3.92(m,2H),3.80(dd,J=11.6,4.5Hz,1H),3.60(t,J=6.1Hz,3H),2.68-2.61(m,1H),2.52(d,J=3.4Hz,3H),2.50(s,1H),2.18(d,J=11.2Hz,1H),2.05(s,3H),1.83(d,J=7.3Hz,2H),1.75(dd,J=13.6,6.7Hz,4H),1.27(s,6H),1.25(s,4H),1.23(s,6H),0.97(s,9H)。LC/MS(ESI+)Calcd for C56H65BrN6O8S(M+H+)m/z,1062.4;Found,1063.4。
9:化合物(2R,4R)-1-((S)-2-(2–((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-(1-(4-(4-甲基-5-基)苯基)环丙基)吡咯烷-2-甲酰胺(9)。
将(3R,5R)-1-((S)-2-(2-((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((1-(4-(4-甲基噻唑-5-基)苯基)环丙基)氨基甲酰基)吡咯烷-3-基乙酸酯(5.0mg,4.70μmol)溶于1ml甲醇,缓慢加入LiOH.H2O(0.9mg,0.02mmol)与0.5ml H2O的混合体系。大约30minTLC监测反应完毕。加入水,DCM萃取。Pre-TLC纯化得到化合物(2R,4R)-1-((S)-2-(2–((6-(4-(((1R,3R)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)己-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-(1-(4-(4-甲基-5-基)苯基)环丙基)吡咯烷-2-甲酰胺3.0mg。收率:84.3%。LC/MS(ESI+)Calcd for C56H65BrN6O8S(M+H+)m/z,1020.4;Found,1021.4。1H NMR(400MHz,CDCl3)δ8.96(s,1H),7.70(s,1H),7.59–7.49(m,2H),7.43(s,1H),7.35(s,2H),7.31(s,1H),7.21(d,J=8.5Hz,1H),7.15(s,1H),6.85(d,J=8.0Hz,1H),6.44–6.34(m,1H),4.66(s,1H),4.58–4.43(m,2H),4.23–4.06(m,3H),4.03–3.89(m,2H),3.62(d,J=16.3Hz,3H),2.53(d,J=13.9Hz,3H),2.47–2.39(m,1H),2.38–2.29(m,1H),2.14(s,2H),2.04(s,1H),1.82(s,4H),1.75(s,10H),1.29(s,6H),1.25(s,9H),1.23(s,6H),1.01(s,7H),0.93–0.78(m,5H).
10:(3R,5S)-1-((S)-2-(2-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基乙炔基)-1H-吡唑-1-基)乙酰氨基)-3,3-二甲基丁酰基)-5-((((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(10)
LC/MS(ESI+)Calcd for C55H59ClN8O7S(M+H+)m/z,1011.4;found 1011.4。1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.47(d,J=7.6Hz,1H),8.32(d,J=8.5Hz,1H),8.13(d,J=6.3Hz,1H),7.91(d,J=8.8Hz,1H),7.76(s,1H),7.70(d,J=7.1Hz,2H),7.59(d,J=8.1Hz,1H),7.41(dd,J=25.1,8.2Hz,4H),7.30(d,J=2.4Hz,1H),7.08(dd,J=8.8,2.4Hz,1H),5.76(s,1H),5.17(s,1H),5.02–4.90(m,3H),4.79(d,J=15.3Hz,2H),4.55(s,1H),4.48(t,J=8.3Hz,1H),4.41–4.31(m,2H),3.90(d,J=11.8Hz,1H),3.74(dd,J=11.6,3.9Hz,1H),3.32(s,1H),2.46(s,3H),1.93(s,3H),1.43–1.34(m,9H),1.16(s,6H),0.98(s,9H).
11:(2S,4R)-1-((S)-2-(2-(4-((2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基乙炔基)-1H-吡唑-1-基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺(11)
LC/MS(ESI+)Calcd for C53H57ClN8O6S(M+H+)m/z,969.4;found 969.4。1H NMR(400MHz,DMSO-d6)δ8.99(d,J=3.0Hz,1H),8.45(d,J=7.6Hz,1H),8.27(d,J=8.8Hz,1H),8.14(d,J=2.8Hz,1H),7.91(dd,J=8.7,3.1Hz,1H),7.77(d,J=2.9Hz,1H),7.73–7.67(m,2H),7.59(d,J=7.9Hz,1H),7.46–7.35(m,4H),7.30(d,J=2.5Hz,1H),7.08(dd,J=8.8,2.5Hz,1H),5.76(d,J=3.1Hz,0H),5.14(d,J=3.3Hz,1H),4.96(dd,J=21.1,4.4Hz,3H),4.81(s,2H),4.58–4.49(m,2H),4.44(d,J=7.9Hz,1H),4.33(s,1H),4.28(s,1H),3.66–3.51(m,2H),2.45(d,J=3.1Hz,3H),2.03(s,1H),1.77(d,J=8.8Hz,1H),1.44–1.36(m,9H),1.16(d,J=2.7Hz,6H),0.96(s,9H).
12:(3R,5S)-1-((S)-2-(2-(2-((3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)丙-2-yn-1-基)氧基)丙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((((S)-1-乙酸(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(12)
LC/MS(ESI+)Calcd for C55H63ClN6O9S(M+H+)m/z,1019.4;found 1019.4。1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.47(d,J=7.6Hz,1H),7.91(d,J=8.8Hz,1H),7.69(d,J=2.9Hz,2H),7.57(d,J=7.9Hz,1H),7.43(d,J=8.3Hz,3H),7.37(s,2H),7.29(d,J=2.3Hz,1H),7.07(dd,J=8.7,2.4Hz,1H),5.20(s,1H),4.90(s,1H),4.79(s,2H),4.54(s,1H),4.49(s,5H),4.32(s,1H),3.99(s,3H),3.75–3.67(m,4H),3.38(s,1H),3.32(s,2H),2.45(s,3H),2.00(s,5H),1.37(dd,J=19.0,4.4Hz,9H),1.23(s,1H),1.15(s,6H),0.95(d,J=7.4Hz,9H).
13:(2S,4R)-1-((S)-2-(2-(2-((3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)丙-2-yn-1-基)氧基)丙氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺(13)
LC/MS(ESI+)Calcd for C53H61ClN6O8S(M+H+)m/z,977.4;found 977.4。1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.47(d,J=7.6Hz,1H),8.32(d,J=8.5Hz,1H),8.13(d,J=6.3Hz,1H),7.91(d,J=8.8Hz,1H),7.76(s,1H),7.70(d,J=7.1Hz,2H),7.59(d,J=8.1Hz,1H),7.41(dd,J=25.1,8.2Hz,4H),7.30(d,J=2.4Hz,1H),7.08(dd,J=8.8,2.4Hz,1H),5.76(s,1H),5.17(s,1H),5.02–4.90(m,3H),4.79(d,J=15.3Hz,2H),4.55(s,1H),4.48(t,J=8.3Hz,1H),4.41–4.31(m,2H),3.90(d,J=11.8Hz,1H),3.74(dd,J=11.6,3.9Hz,1H),3.32(s,1H),2.46(s,3H),1.93(s,3H),1.43–1.34(m,9H),1.16(s,6H),0.98(s,9H).
14:(3R,5S)-1-((2S)-2-(2-(2-((3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)丙-2-yn-1-基)氧基)丙氧基)乙酰氨基)-3,3-二甲基丁酰基)-5-((((S)-1-乙酸(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯(14);
LC/MS(ESI+)Calcd for C56H65ClN6O9S(M+H+)m/z,1033.4;found 1033.4。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.48(d,J=6.7Hz,1H),7.91(d,J=8.8Hz,1H),7.70(d,J=7.6Hz,2H),7.58(t,J=6.0Hz,1H),7.40(dd,J=27.2,7.6Hz,4H),7.29(d,J=2.3Hz,1H),7.07(dd,J=8.8,2.3Hz,1H),5.20(s,1H),4.89(s,1H),4.79(s,2H),4.56–4.42(m,5H),4.32(s,1H),4.00(d,J=6.7Hz,2H),3.93–3.85(m,1H),3.81–3.70(m,2H),3.57(s,3H),3.41–3.37(m,1H),3.20–3.10(m,1H),2.69(s,9H),2.45(s,3H),2.31–2.21(m,1H),2.00(s,3H),1.98(s,1H),1.39(s,5H),1.36(dd,J=6.9,3.2Hz,3H),1.26(t,J=6.4Hz,6H),1.14(d,J=6.0Hz,7H),0.97(s,9H).
15:(2S,4R)-1-((2S)-2-(2-(2-((3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)丙-2-yn-1-基)氧基)丙氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺(15)
LC/MS(ESI+)Calcd for C54H63ClN6O8S(M+H+)m/z,991.4;found 991.4。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.46(d,J=6.7Hz,1H),7.90(d,J=8.8Hz,1H),7.71(d,J=7.6Hz,2H),7.58(t,J=6.0Hz,1H),7.42(dd,J=27.2,7.6Hz,4H),7.31(d,J=2.3Hz,1H),7.07(dd,J=8.8,2.3Hz,1H),5.21(s,1H),4.89(s,1H),4.79(s,2H),4.57–4.43(m,5H),4.32(s,1H),4.00(d,J=6.7Hz,2H),3.92–3.84(m,1H),3.81–3.70(m,2H),3.41–3.37(m,1H),3.20–3.10(m,1H),2.69(s,9H),2.45(s,3H),2.31–2.21(m,1H),2.00(s,3H),1.98(s,1H),1.39(s,5H),1.38(dd,J=6.9,3.2Hz,3H),1.26(t,J=6.4Hz,6H),1.11(d,J=6.0Hz,7H),0.97(s,9H).
16:(2R,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)氨基)苯基)氧)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯-2-氨甲酰的合成(16)
1.((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯的合成
将(3-羟基-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯(4700mg,19.31mmol)溶于100mL DMF,尔后对体系进行氩气置换操作,反复8次,确保体系中的惰性气体氛围。再将体系移置于冰水浴中降温冷却搅拌,当体系内温降至0℃左右,开始缓慢地向体系中加入NaH(60%)(1600mg,40.55mmol),完毕,体系在0℃保温搅拌1h。随后向体系中滴加50mL溶有2-三氟甲基-4-氟苯腈(1700mg,11.00mmol)的DMF溶液,控制滴加速度,使体系内温最高不超过5℃。滴毕,体系继续在0℃反应。2h后,取样点板,TLC显示反应结束。搅拌下将体系缓慢倒入冰水中,析出大量固体(有黏性),滤饼用乙酸乙酯溶解,无水硫酸钠干燥,过滤浓缩,得粗品,后经柱层析分离纯化得类白色固体((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯(3700mg)。收率:46%。LC/MS(ESI+)Calcd forC21H27F3N2O3(M+H)+m/z,412.2;Found:357.1。
2.4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-(三氟甲基)苯腈的合成
称取((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯(650mg,1.58mmol)置于25mL单颈圆底烧瓶中,同时加入二氯甲烷(15mL),室温搅拌。随后向体系中加入三氟乙酸(3mL),完毕,体系在室温搅拌反应。3h后,取样点板,TLC显示原料已经消耗完毕。旋蒸除去溶剂以及过量的三氟乙酸,并使用二氯甲烷反复旋带除去残留的三氟乙酸,得类白色固体4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-(三氟甲基)苯腈三氟乙酸盐。不经进一步纯化,直接用于下步反应中。
3.2-((((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基)甲基)-4-硝基苯甲酸甲酯的合成
向装有4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-(三氟甲基)苯腈三氟乙酸盐的25mL单颈圆底烧瓶中加入DMF(10mL),室温搅拌溶解澄清。随后,依次向体系中加入碳酸钾(437mg,3.16mmol),2-(溴甲基)-4-硝基苯甲酸甲酯(433mg,1.58mmol)。完毕,体系在100℃的油浴中搅拌反应。1h后,取样点板,TLC显示原料已经消耗完毕。向体系中加入乙酸乙酯(15mL)和水(10mL),剧烈搅拌,3min后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,后对其进行柱层析分离纯化得2-((((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基)甲基)-4-硝基苯甲酸甲酯(532mg)。收率(两步):67%。
LC/MS(ESI+)Calcd for C25H26F3N3O5(M+H)+m/z,505.1;Found:506.3。
4.4-((1r,3r)-2,2,4,4-四甲基-3-(5-硝基-1-氧代异吲哚啉-2-yl)环氧丁氧基)-2-(三氟甲基)苯腈
称取2-((((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)氨基)甲基)-4-硝基苯甲酸甲酯(532mg,1.05mmol)于25mL单颈圆底烧瓶中,并加入甲苯(10mL),室温搅拌。随后,向体系中加入三乙胺(319mg,3.15mmol)。完毕,将体系移置于110℃油浴中继续升温加热回流搅拌反应过夜。翌日,取样点板,TLC显示反应结束。待体系冷却至室温后,旋蒸除去溶剂,得粗品。对其使用正己烷(20mL)和乙酸乙酯(1mL)在室温进行打浆操作。30min后,对该体系进行抽滤操作,滤饼用正己烷(30mL)淋洗,干燥,得白色固体4-((1r,3r)-2,2,4,4-四甲基-3-(5-硝基-1-氧代异吲哚啉-2-yl)环氧丁氧基)-2-(三氟甲基)苯腈(395mg)。收率:79%。
LC/MS(ESI+)Calcd for C24H22F3N3O4(M+H)+m/z,473.1;Found:474.3。
5.4-((1r,3r)-3-(5-氨基-1-氧代异吲哚啉-2-yl)-2,2,4,4-四甲基环氧丁氧基)-2-(三氟甲基)苯腈
称取4-((1r,3r)-2,2,4,4-四甲基-3-(5-硝基-1-氧代异吲哚啉-2-yl)环氧丁氧基)-2-(三氟甲基)苯腈(395mg,0.83mmol)于25mL单颈圆底烧瓶中,并加入冰醋酸(12mL),室温搅拌。随后向体系中加入还原铁粉(1400mg,24.90mmol),完毕,将体系移置于65℃的油浴中继续升温加热搅拌反应。30min后,取样点板,TLC显示原料已经消耗完毕,停止加热。趁热对体系进行过滤操作,滤饼用冰醋酸(36mL)淋洗。收集滤液,旋蒸除去醋酸,得粗品。对其使用正己烷(5mL)和乙酸乙酯(2mL)在室温进行打浆操作。20min后,对该体系进行抽滤操作,滤饼用正己烷(5mL)淋洗,干燥,得类白色固体4-((1r,3r)-3-(5-氨基-1-氧代异吲哚啉-2-yl)-2,2,4,4-四甲基环氧丁氧基)-2-(三氟甲基)苯腈(324mg)。收率:88%。
LC/MS(ESI+)Calcd for C24H24F3N3O2(M+H)+m/z,443.1;Found:444.2。
6.2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环氧丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸叔丁酯的合成
称取4-((1r,3r)-3-(5-氨基-1-氧代异吲哚啉-2-yl)-2,2,4,4-四甲基环氧丁氧基)-2-(三氟甲基)苯腈(150mg,0.34mmol)置于50mL单颈圆底烧瓶中,依次加,2-((5-氧代戊基)氧)乙酸叔丁酯(73mg,0.34mmol),甲醇(15mL)以及醋酸(61mg,1.02mmol),室温搅拌溶解澄清。15min后,向体系中加入NaBH3CN(86mg,1.36mmol),完毕,体系在室温搅拌反应过夜。翌日,取样点板,原料基本消耗完。旋蒸除去溶剂后,向体系中加入乙酸乙酯(20mL)和水(10mL),剧烈搅拌,3min后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,并通过Pre-TLC分离纯化得类白色固体2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环氧丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸叔丁酯(80mg)。收率:37%。
LC/MS(ESI+)Calcd for C35H44F3N3O5(M+H)+m/z,643.3;Found:644.3。
7.2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸的合成
称取2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环氧丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸叔丁酯(38mg,0.06mmol)置于25mL单颈圆底烧瓶中,同时加入二氯甲烷(10mL),室温搅拌。随后向体系中加入三氟乙酸(1mL),完毕,体系在室温搅拌反应。3h后,取样点板,TLC显示原料已经消耗完毕。旋蒸除去溶剂以及过量的三氟乙酸,并使用二氯甲烷反复旋带除去残留的三氟乙酸,得类白色固体2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸。不经进一步纯化,直接用于下步反应中。
8.(2R,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)氨基)苯基)氧)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯-2-氨甲酰的合成
向装有2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)氨基)戊基)氧)乙酸的25mL单颈圆底烧瓶中依次加入HATU(34mg,0.09mmol),DMF(5mL),VHL(OAc)(39mg,0.07mmol)以及二异丙基乙胺(23mg,0.18mmol),尔后体系在室温搅拌反应过夜。翌日取样点板,TLC显示原料消耗完毕。向体系中加入乙酸乙酯(15mL)和水(10mL),剧烈搅拌,3min后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得类白色固体(2R,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(4-氰基-3-(三氟甲基)苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-yl)氨基)苯基)氧)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-1-(4-(4-甲基噻唑-5-yl)苯基)乙基)吡咯-2-氨甲酰(31mg)。收率(两步):52%。
LC/MS(ESI+)Calcd for C54H66F3N7O7S(M+H)+m/z,1013.4;Found:1014.2。
1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.44(d,J=7.8Hz,1H),8.10(d,J=8.7Hz,1H),7.46–7.31(m,10H),6.69–6.57(m,2H),6.39–6.27(m,1H),5.14(s,1H),4.92–4.84(m,1H),4.62(s,2H),4.58(s,1H),4.55(d,J=9.8Hz,1H),4.45(t,J=8.3Hz,1H),3.92(d,J=2.2Hz,2H),3.60–3.56(dd,J=4.8,1.8Hz,2H),3.50(t,J=6.3Hz,3H),3.13–3.05(m,2H),2.45(s,3H),2.11–2.01(m,1H),1.80–1.73(m,1H),1.66–1.54(m,4H),1.50–1.41(m,2H),1.38(s,6H),1.34(d,J=6.9Hz,2H)1.13(s,6H),0.94(s,9H).
17.(3R,5S)-1-((2S)-2-(2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰)吡咯烷-3-基乙酸酯
1.合成中间体:1.4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-羧酸叔丁酯
称取6-[4-(叔丁氧羰酰)哌嗪-1-基]烟酸(130mg,0.42mmol)置于25mL单颈圆底烧瓶中,同时加入DMF(8mL),室温搅拌溶解澄清。随后向体系中依次加入EDCI(200mg,1.05mmol),HOBt(85mg,0.63mmol),DMAP(5mg,0.04mmol)以及三乙胺(106mg,1.05mmol),完毕,体系在室温搅拌反应5min。尔后,向体系中加入按文献方法制备得到的4-((3aR,4R,7R,7aS)-5-氨基-4,7-二甲基-1,3-二氧六氢-1氢-4,7-环氧异吲哚-2(3氢)-基)-2-氯苯腈(145mg,0.42mmol),任体系在室温搅拌反应过夜。翌日,TLC检测反应结束。向体系中加入乙酸乙酯(20mL)和水(15mL),剧烈搅拌,5min后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,并通过Pre-TLC分离纯化得4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-羧酸叔丁酯(191mg)。收率:71%。LC/MS(ESI+)Calcd for C32H35ClN6O6(M+H)+m/z,635.1;Found:634.7。
2.N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)-6-(哌嗪-1-基)烟酰胺三氟醋酸盐的合成
称取4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-羧酸叔丁酯(175mg,0.28mmol)置于25mL单颈圆底烧瓶中,同时加入二氯甲烷(10mL),室温搅拌。随后向体系中加入三氟乙酸(1mL),完毕,体系在室温搅拌反应。3.5h后,取样点板,TLC显示原料已经消耗完毕。旋蒸除去溶剂以及过量的三氟乙酸,并使用二氯甲烷反复旋带除去残留的三氟乙酸,得类白色固体N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)-6-(哌嗪-1-基)烟酰胺三氟醋酸盐。不经进一步纯化,直接用于下步反应中。
3.4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-羧酸叔丁酯的合成
向装有N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)-6-(哌嗪-1-基)烟酰胺三氟醋酸盐的25mL单颈圆底烧瓶中分别加入1-Boc-4溴甲基哌啶(86mg,0.31mmol),碳酸钾(155mg,1.12mmol),碘化钠(42mg,0.28mmol)以及乙腈(10mL),室温搅拌溶解。尔后对体系进行抽真空,通氩气的操作反复5次,确保体系中的惰性气体氛围。完毕,将体系移置于油浴中升温加热回流反应过夜。15h后取样点板,TLC显示反应结束。旋蒸除去溶剂,再向体系中加入乙酸乙酯(15mL)和水(10mL),剧烈搅拌,3min后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,并通过Pre-TLC分离纯化得4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-羧酸叔丁酯(124mg)。收率(两步):61%。LC/MS(ESI+)Calcd for C38H46ClN7O6(M+H)+m/z,732.3;Found:732.2。
4.N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)-6-(4-(哌啶-4-基甲基)哌嗪-1-基)烟酰胺三氟乙酸盐的合成
称取4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-羧酸叔丁酯(124mg,0.16mmol)置于25mL单颈圆底烧瓶中,同时加入二氯甲烷(8mL),室温搅拌。随后向体系中加入三氟乙酸(1mL),完毕,体系在室温搅拌反应。3h后,取样点板,TLC显示反应已经结束。旋蒸除去溶剂以及过量的三氟乙酸,并使用二氯甲烷反复旋带除去残留的三氟乙酸,得类白色固体N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)-6-(4-(哌啶-4-基甲基)哌嗪-1-基)烟酰胺三氟乙酸盐。不经进一步纯化,直接用于下步反应。
5.2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸叔丁酯的合成
向装有N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)-6-(4-(哌啶-4-基甲基)哌嗪-1-基)烟酰胺三氟乙酸盐的25mL单颈圆底烧瓶中依次加入溴乙酸叔丁酯(62mg,0.32mmol),二氯甲烷(10mL)以及二异丙基乙胺(83mg,0.64mmol),完毕,体系在室温搅拌反应过夜。翌日取样点板,TLC显示反应结束。向体系中加入二氯甲烷(10mL)和水(10mL),剧烈搅拌,3min后静置分层,水层用二氯甲烷反萃(5mL*3),合并有机相,依次用水(5mL*3),饱和食盐水(10mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,后经Pre-TLC分离纯化得2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸叔丁酯(74mg)。收率(两步):59%。
LC/MS(ESI+)Calcd for C39H48ClN7O6(M+H)+m/z,746.3;Found:745.2。
6.2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸的合成
称取2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸叔丁酯(40mg,0.05mmol)置于25mL单颈圆底烧瓶中,同时加入二氯甲烷(5mL),室温搅拌。随后向体系中加入三氟乙酸(1mL),完毕,体系在室温搅拌反应。3h后,取样点板,TLC显示反应已经结束。旋蒸除去溶剂以及过量的三氟乙酸,并使用二氯甲烷反复旋带除去残留的三氟乙酸,得类白色固体2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸。不经进一步纯化,直接用于下步反应中。
7.(3R,5S)-1-((2S)-2-(2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰)吡咯烷-3-基乙酸酯
向装有2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酸的25mL单颈圆底烧瓶中依次加入HATU(29mg,0.08mmol),DMF(5mL),VHL(OAc)(36mg,0.06mmol)以及二异丙基乙胺(19mg,0.15mmol),尔后体系在室温搅拌反应过夜。翌日取样点板,TLC显示原料消耗完毕。向体系中加入乙酸乙酯(15mL)和水(10mL),剧烈搅拌,后静置分层,水层用乙酸乙酯反萃(10mL*3),合并有机相,依次用水(10mL*3),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,真空浓缩得粗品,后经Pre-TLC分离纯化得(3R,5S)-1-((2S)-2-(2-(4-((4-(5-(((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)乙酰氨基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰)吡咯烷-3-基乙酸酯(34mg)。收率(两步):55%。LC/MS(ESI+)Calcd for C60H72ClN11O9S(M+H)+m/z,1158.8;Found:1157.6。
1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.46(d,J=7.7Hz,1H),8.30(d,J=7.7Hz,1H),8.13(d,J=8.4Hz,1H),7.77(dd,J=8.4,5.3Hz,3H),7.70(d,J=9.4Hz,1H),7.53(dd,J=8.4,1.8Hz,1H),7.40(dd,J=28.8,8.2Hz,4H),6.98(d,J=8.5Hz,2H),5.20(s,1H),4.94–4.86(m,1H),4.49–4.43(m,1H),4.41–4.30(m,2H),3.88(d,J=12.2Hz,1H),3.76(dd,J=11.1,4.0Hz,1H),3.47(d,J=7.2Hz,1H),3.29–3.25(m,7H),2.86–2.76(m,2H),2.46(s,3H),2.35–2.05(m,7H),2.00(s,3H),1.94–1.85(m,2H),1.79–1.70(m,2H),1.49(s,3H),1.42(s,3H),1.38(d,J=7.0Hz,3H),1.24(s,2H),1.17–1.10(m,3H),0.95(s,9H).
18.N-((3aR,4R,7R,7aS)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1H-4,7-环氧异吲哚啉-5-基)-6-(4-((1-(2-(((S)-1-((2S,4R)-4-羟基-2-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)氨基)-2-氧乙基)哌啶-4-基)甲基)哌嗪-1-基)烟酰胺的合成
LC/MS(ESI+)Calcd for C58H70ClN11O8S(M+H)+m/z,1116.8;Found:1116.6。
1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.66(d,J=2.2Hz,1H),8.45(d,J=7.7Hz,1H),8.37(d,J=7.2Hz,1H),8.13(d,J=8.4Hz,1H),7.99(dd,J=9.2,2.2Hz,1H),7.76(d,J=1.8Hz,1H),7.53(dd,J=8.4,1.9Hz,1H),7.40(dd,J=28.7,8.2Hz,4H),6.88(d,J=9.0Hz,1H),5.13(d,J=3.1Hz,1H),4.92–4.86(m,1H),4.50(d,J=9.8Hz,1H),4.44(t,J=8.1Hz,1H),4.38–4.31(m,1H),4.28(dt,J=9.2,5.6Hz,1H),3.61–3.52(m,7H),3.48(d,J=7.2Hz,1H),2.84–2.75(m,3H),2.46(s,3H),2.11–1.94(m,9H),1.89(dd,J=12.8,5.2Hz,2H),1.79–1.69(m,3H),1.49(s,3H),1.43(s,3H),1.38(d,J=7.0Hz,3H),1.18–0.97(m,5H),0.94(s,9H).
19:(3S,5R)-1-((2R)-2-(2-((5-((4-(((3aS,4S,7S,7aR)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)-3-氟苯基)氨基)戊基)氧)乙酰氨基)-3,3-二甲基丁酰基)-5-(((R)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰)吡咯烷-3-基乙酸酯
LC/MS(ESI+)Calcd for C56H64ClFN8O10S(M+H)+m/z,1095.7;Found:1095.2。
1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.44(dd,J=11.8,7.4Hz,2H),8.13(d,J=8.5Hz,1H),7.92(s,1H),7.76(d,J=1.7Hz,1H),7.69(dd,J=8.7,6.6Hz,1H),7.53(dd,J=8.4,1.5Hz,1H),7.39(dt,J=14.0,7.0Hz,5H),6.47(dd,J=12.9,2.3Hz,1H),6.40(td,J=8.2,2.4Hz,1H),5.20(dd,J=4.0,3.1Hz,1H),4.94–4.85(m,1H),4.44(dd,J=16.0,8.7Hz,2H),4.31(td,J=12.6,5.8Hz,1H),3.95–3.84(m,3H),3.76(dd,J=11.5,3.5Hz,1H),3.48(m,3H),3.11(dd,J=11.6,6.0Hz,2H),2.69(s,3H),2.45(s,3H),2.30–2.21(m,1H),2.11(t,J=12.0Hz,1H),1.90(dd,J=13.4,4.6Hz,1H),1.65–1.55(m,4H),1.49(s,3H),1.46–1.39(m,5H),1.36(d,J=7.1Hz,3H),1.23(s,2H),0.94(s,9H).
20:(2R,4S)-1-((2R)-2-(2-((5-((4-(((3aS,4S,7S,7aR)-2-(3-氯-4-苯腈)-4,7-二甲基-1,3-二氧八氢-1氢-4,7-环氧异吲哚啉-5-基)氨基甲酰)-3-氟苯基)氨基)戊基)氧)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-氨甲酰
LC/MS(ESI+)Calcd for C54H62ClFN8O9S(M+H)+m/z,1053.6;Found:1053.5。
1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.43(t,J=6.6Hz,2H),8.13(d,J=8.4Hz,1H),7.92(s,1H),7.76(d,J=1.7Hz,1H),7.69(dd,J=8.2,6.9Hz,1H),7.53(dd,J=8.5,1.7Hz,1H),7.48–7.25(m,6H),6.52–6.36(m,2H),5.13(d,J=3.5Hz,1H),4.89(m,1H),4.54(d,J=9.6Hz,1H),4.45(t,J=7.9Hz,1H),4.28(m,1H),3.92(s,2H),3.60–3.55(m,2H),3.49(dd,J=12.5,7.0Hz,3H),3.30(s,3H),3.11(qd,J=7.3,1.4Hz,2H),2.45(s,3H),2.16–2.01(m,2H),1.93–1.86(m,1H),1.81–1.72(m,1H),1.67–1.54(m,4H),1.49(s,3H),1.43(s,3H),1.36(d,J=6.8Hz,3H),0.93(s,9H).
21:(2S,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-氢-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(21)
1.合成2-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酸叔丁酯
将化合物4-((1R,3R)-3-(5-溴-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(1-1)(189mg,0.4mmol)和2-(戊基-4-炔-1-氧基)乙酸叔丁酯(119mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到177mg中间体1-2,收率:75%。质谱(ES):m/z=591[M+H]+。
2.合成中间体:2-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酸
将化合物2-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酸叔丁酯(中间体1-2)(118mg,0.2mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
3.合成最终产物:(2S,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-氢-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-炔-1-基)氧基)乙酸(中间体1-3)(53.5mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.05mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到52mg最终产物,收率:54%。质谱(ES):m/z=961[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.51-8.44(m,1H),7.91(s,1H),7.63(d,J=9.9Hz,2H),7.54-7.33(m,6H),7.29(s,1H),7.11-7.03(m,1H),5.25-5.17(m,2H),4.95-4.86(m,1H),4.77(s,2H),4.54(s,3H),4.31(s,1H),3.98(s,3H),3.82-3.73(m,1H),3.63(s,2H),2.46(s,3H),2.32-2.21(m,2H),1.86(s,2H),1.39(s,8H),1.24(s,3H),1.15(s,6H),0.96(s,9H).
22:(2S,4R)-1-((S)-2-(2-((5-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(三氘代甲基)苯甲酰胺)-4-炔基戊基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((s)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
合成中间体:((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酸叔丁酯
将化合物((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯(1.89g,5mmol)溶于5mL DMF中,冰浴条件下加入钠氢(0.4g,10mmol),室温搅拌0.5h,加入氘代碘甲烷(1.09g,7.5mmol),继续搅拌反应3h,反应完成后加入水,并用乙酸乙酯萃取,有机相减压蒸馏除去溶剂,粗品经柱层析纯化得到1.74g中间体2-2,收率88%。质谱(ES):m/z=397[M+H]+。
合成中间体:((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酸
将化合物((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酸叔丁酯(中间体2-2)(396mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
合成中间体:N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-氘代甲基苯甲酰胺
将化合物((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酸(中间体2-3)(296mg,1mmol)和对碘苯甲酸(248mg,1mmol)溶于2mL DMF,DIPEA(258mg,2mmol),HATU(570mg,1.5mmol)后,室温搅拌3h,反应完成后加入水并用乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(PE:EA=3:1),得到395mg中间体2-4,收率:75%。质谱(ES):m/z=526[M+H]+。
合成中间体:2-((5-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)戊基-5-炔-1-基)氧基)乙酸叔丁酯
将化合物N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-氘代甲基苯甲酰胺(中间体2-4)(210mg,0.4mmol)和2-(戊基-4-炔-1-氧基)乙酸叔丁酯(119mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到203mg中间体2-5,收率:85%。质谱(ES):m/z=596[M+H]+。合成中间体:2-((5-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)戊基-5-炔-1-基)氧基)乙酸
将化合物2-((5-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)戊基-5-炔-1-基)氧基)乙酸叔丁酯(中间体2-5)(59.6mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
将化合物2-((5-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)戊基-5-炔-1-基)氧基)乙酸(中间体2-6)(54mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.05mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到49mg最终产物HC-2797-01,收率:51%。质谱(ES):m/z=966[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.46(d,J=7.6Hz,1H),7.87(d,J=8.7Hz,1H),7.45(dd,J=12.0,8.1Hz,5H),7.41-7.34(m,4H),7.25(s,1H),7.03(d,J=8.4Hz,1H),5.16(d,J=3.3Hz,1H),4.96-4.87(m,1H),4.67(d,J=7.9Hz,1H),4.56(d,J=9.5Hz,1H),4.46(t,J=8.1Hz,1H),4.29(s,1H),3.98(s,2H),3.62(dd,J=13.7,7.8Hz,4H),2.58-2.49(m,8H),2.12-2.02(m,1H),1.90-1.74(m,3H),1.48(d,J=6.9Hz,1H),1.40-1.32(m,9H),1.23(s,3H),0.94(s,9H).
23:(2S,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-5-炔基己基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.合成中间体:2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)己基-5-炔-1-基)氧基)乙酸叔丁酯
将化合物N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-氘代甲基苯甲酰胺(中间体2-4)(210mg,0.4mmol)和2-(己基-5-炔-1-氧基)乙酸叔丁酯(127mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到205mg中间体4-1,收率:84%。质谱(ES):m/z=610[M+H]+。
2.合成中间体:2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)己基-5-炔-1-基)氧基)乙酸
将化合物2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)己基-5-炔-1-基)氧基)乙酸叔丁酯(中间体4-1)(61mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
3.合成最终产物HC-2799-01:(2S,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-5-炔基己基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)己基-5-炔-1-基)氧基)乙酸(中间体4-2)(55mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到53mg最终产物HC-2799-01,收率:54%。质谱(ES):m/z=980[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.45(d,J=7.7Hz,1H),7.87(d,J=8.6Hz,1H),7.38(ddd,J=48.2,28.5,19.8Hz,9H),7.25(s,1H),7.03(d,J=9.0Hz,1H),5.15(d,J=3.4Hz,1H),4.95-4.85(m,1H),4.77-4.61(m,1H),4.55(d,J=9.6Hz,1H),4.46(t,J=8.2Hz,1H),4.29(s,1H),3.92(d,J=16.9Hz,2H),3.56(dd,J=14.7,9.1Hz,4H),2.48(d,J=21.7Hz,11H),2.05(d,J=8.4Hz,1H),1.82-1.59(m,4H),1.42-1.17(m,11H),0.94(s,9H).
24:(2S,4R)-1-((S)-2-(2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-6-炔基庚基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.合成中间体:2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)庚基-6-炔-1-基)氧基)乙酸叔丁酯
将化合物N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-氘代甲基苯甲酰胺(中间体2-4)(210mg,0.4mmol)和2-(庚基-6-炔-1-氧基)乙酸叔丁酯(136mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到212mg中间体6-1,收率:85%。质谱(ES):m/z=624[M+H]+。
2.合成中间体:2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)庚基-6-炔-1-基)氧基)乙酸
将化合物2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)庚基-6-炔-1-基)氧基)乙酸叔丁酯(中间体6-1)(62mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
3.合成最终产物24:(2S,4S)-1-((S)-2-(2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-6-炔基庚基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((s)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-((7-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)氨基甲酰)苯基)庚基-6-炔-1-基)氧基)乙酸(中间体6-2)(57mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到50mg最终产物HC-2801-01,收率:50%。质谱(ES):m/z=994[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.45(d,J=7.7Hz,1H),7.87(d,J=8.6Hz,1H),7.38(ddd,J=48.2,28.5,19.8Hz,9H),7.25(s,1H),7.03(d,J=9.0Hz,1H),5.15(d,J=3.4Hz,1H),4.95-4.85(m,1H),4.77-4.61(m,1H),4.55(d,J=9.6Hz,1H),4.46(t,J=8.2Hz,1H),4.29(s,1H),3.92(d,J=16.9Hz,2H),3.56(dd,J=14.7,9.1Hz,4H),2.48(d,J=21.7Hz,11H),2.05(d,J=8.4Hz,1H),1.82-1.59(m,4H),1.42-1.17(m,13H),0.94(s,9H).
25:N-(5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊基-4-炔-1-基)-N-((S)-1-((2S,4R)-4-羟基-2-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)环丙烷-1,1-二甲酰胺
1.合成中间体:2-(戊-4-烯-1-基)异吲哚啉-1,3-二酮
将戊-4-炔-1-醇(4.2g,0.05mol)溶于30mL DCM中,加入TEA(7.6g,0.075mol),冰浴条件下滴加甲基磺酰氯(6.9g,0.06mol),搅拌反应5h。反应完成后,加入25mL水和30mL二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品未进纯化直接用于下一步反应。
将上述产物(3.2g,0.02mol)溶于20mL DMF中,加入邻苯二甲酰亚胺钾盐(5.6g,0.03mol),反应液温度升高到80℃,搅拌反应6h。反应完成后,加入30mL水和90mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用柱层析纯化(PE:EA=3:1),得到3.9g中间体8-3,收率:91%。质谱(ES):m/z=214[M+H]+。
2.合成中间体:戊-4-炔-1-胺
将上述产物(2.1g,0.01mol)溶于30mL无水乙醇中,加入水合肼(0.75g,0.015mol),反应液温度升高到80℃,搅拌反应5h。反应完成后,待反应液冷却,过滤除去不溶物,向滤液中加入30mL水和90mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂,粗品未进纯化直接用于下一步反应。
3.合成中间体:1-(4-戊炔-1-氨甲酰)环丙烷-1-羧酸甲酯
将上述产物戊炔胺(830mg,10mmol)和1-甲氧基羰基环丙烷-1-甲酸(1.44g,10mmol)溶于2mL二氯甲烷中,DIPEA(3.9g,30mmol),HATU(7.6g,20mmol)后,室温搅拌4h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(PE:EA=3:1),得到1.79g中间体8-5,收率:85%。质谱(ES):m/z=210[M+H]+。
4.合成中间体:1-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-环-1-基)氨甲酰)环丙烷-1-羧酸甲酯
将化合物4-((1R,3R)-3-(5-溴-1-氧代异丁醇-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(1-1)(189mg,0.4mmol)和1-(4-戊炔-1-氨甲酰)环丙烷-1-羧酸甲酯(126mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=2:1),得到188mg中间体8-6,收率:78%。质谱(ES):m/z=602[M+H]+。
5.合成中间体:1-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-环-1-基)氨甲酰)环丙烷-1-羧酸
将化合物1-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-环-1-基)氨甲酰)环丙烷-1-羧酸甲酯(中间体8-6)(60mg,0.1mmol)溶于2mL无水甲醇中,加入氢氧化锂(9.6mg,0.4mmol),室温搅拌5h。反应完成后,向滤液中加入10mL水和20mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂,粗品未进纯化直接用于下一步反应。
6.合成最终产物25:N-(5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊基-4-炔-1-基)-N-((S)-1-((2S,4R)-4-羟基-2-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)环丙烷-1,1-二甲酰胺
将化合物1-((5-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)戊-4-环-1-基)氨甲酰)环丙烷-1-羧酸(中间体8-7)(59mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到55mg最终产物HC-2803-01,收率:54%。质谱(ES):m/z=1014[M+H]+。1H NMR(400MHz,DMSO-d6)δ9.40(d,J=8.7Hz,1H),8.98(s,1H),8.42(d,J=7.7Hz,1H),7.90(d,J=8.8Hz,1H),7.82(s,1H),7.66(d,J=7.9Hz,1H),7.61(s,1H),7.51(d,J=7.8Hz,1H),7.43(d,J=8.3Hz,2H),7.37(d,J=8.2Hz,2H),7.28(d,J=2.3Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),5.13(d,J=3.5Hz,1H),4.90(s,1H),4.77(s,2H),4.59-4.38(m,3H),4.30(d,J=10.6Hz,2H),3.59(s,2H),3.23(d,J=6.2Hz,2H),2.49–2.43(m,4H),2.08-1.97(m,1H),1.83-1.68(m,3H),1.42-1.34(m,8H),1.32-1.21(m,8H),1.15(s,4H),0.94(d,J=11.9Hz,9H).
26:(2S,4R)-1-((S)-2-(2-((4-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-3-炔基丁基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.合成中间体:2-((4-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-3-炔-1-丁基)氧基)乙酸叔丁酯
将化合物N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-氘代甲基苯甲酰胺(210mg,0.4mmol)和2-(丁基-3-炔-1-氧基)乙酸叔丁酯(111mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到203mg产物,收率:87%。质谱(ES):m/z=582[M+H]+。
2.合成中间体:2-((4-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-3-炔-1-丁基)氧基)乙酸
将化合物2-((4-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-3-炔-1-丁基)氧基)乙酸叔丁酯(58mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
3.合成最终产物26:(2S,4R)-1-((S)-2-(2-((4-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(氘代甲基)苯甲酰胺)-3-炔基丁基-氧基乙酰氨基-3,3-二甲基丁烷)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-((4-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-3-炔-1-丁基)氧基)乙酸(53mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到50mg最终产物26,收率:52%。质谱(ES):m/z=952[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.46(d,J=7.8Hz,1H),7.88(d,J=8.5Hz,1H),7.55-7.35(m,9H),7.26(s,1H),7.08-7.00(m,1H),5.38(s,1H),5.15(s,1H),4.92(s,1H),4.56(d,J=9.3Hz,2H),4.46(s,2H),4.29(s,1H),4.04(s,3H),3.65(d,J=40.9Hz,6H),2.77(s,2H),2.12-1.93(m,2H),1.83-1.67(m,2H),1.36(d,J=12.4Hz,9H),0.92(s,12H).
27:(2R,4R)-1-((R)-2-(2-((4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丁醇-5-基)-3-丁炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.合成中间体:2-((4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-3-丁炔-1-基)氧)乙酸叔丁酯
将化合物4-((1R,3R)-3-(5-溴-1-氧代异丁醇-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(1-1)(189mg,0.4mmol)和2-(丁基-3-炔-1-氧基)乙酸叔丁酯(119mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到180mg中间体12-1,收率:78%。质谱(ES):m/z=577[M+H]+。
2.合成中间体:2-((4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-3-丁炔-1-基)氧)乙酸
将化合物2-((4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-3-丁炔-1-基)氧)乙酸叔丁酯(中间体12-1)(58mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
3.合成最终产物27:(2R,4R)-1-((R)-2-(2-((4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丁醇-5-基)-3-丁炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-((4-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-3-炔-1-丁基)氧基)乙酸(中间体10-2)(52mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到53mg最终产物27,收率:56%。质谱(ES):m/z=947[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.51-8.44(m,1H),7.91(s,1H),7.63(d,J=9.9Hz,2H),7.54-7.33(m,6H),7.29(s,1H),7.11-7.03(m,1H),5.25-5.17(m,2H),4.95-4.86(m,1H),4.77(s,2H),4.54(s,3H),4.31(s,1H),3.98(s,3H),3.82-3.73(m,1H),3.63(s,2H),2.46(s,3H),2.32-2.21(m,2H),1.86(s,2H),1.39(s,8H),1.24(s,3H),1.15(s,6H),0.96(s,9H).
28:N-(5-(4-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲基-d3)氨甲酰)苯基)-4-戊炔-1-基)-N-((S)-1-((2S,4R)-4-羟基-2-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)环丙烷-1,1-二甲酰胺
得到51mg最终产物28,收率:50%。质谱(ES):m/z=1019[M+H]+。1H NMR(400MHz,DMSO-d6)δ9.39(d,J=8.8Hz,1H),8.99(s,1H),8.42(d,J=7.7Hz,1H),7.89-7.80(m,2H),7.49-7.41(m,4H),7.37(d,J=8.0Hz,2H),7.26(s,1H),7.04(s,1H),5.12(s,1H),4.92(d,J=7.0Hz,1H),4.78-4.60(m,2H),4.46(dd,J=21.3,8.4Hz,3H),4.29(s,1H),3.55(d,J=29.2Hz,5H),3.22(s,2H),3.00-2.83(m,1H),2.46(s,5H),2.07-1.95(m,1H),1.84-1.64(m,3H),1.41-1.31(m,9H),1.28(d,J=9.5Hz,4H),0.94(d,J=10.6Hz,12H).
29:(2S,4R)-1-((S)-2-(2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丁醇-5-基)庚-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-N-((S)-1-(4-(3,5-二甲基噻唑-4-基)苯基)乙基)-4-羟基吡咯烷-2-甲酰胺
得到51mg最终产物HC-2820-01,收率:52%。质谱(ES):m/z=975[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.51-8.44(m,1H),7.91(s,1H),7.63(d,J=9.9Hz,2H),7.54-7.33(m,6H),7.29(s,1H),7.11-7.03(m,1H),5.25-5.17(m,2H),4.95-4.86(m,1H),4.77(s,2H),4.54(s,3H),4.31(s,1H),3.98(s,3H),3.82-3.73(m,1H),3.63(s,2H),2.46(s,3H),2.32-2.21(m,2H),1.76(m,2H),1.52(m,2H),1.39(s,8H),1.24(s,3H),1.15(s,6H),0.96(s,9H).
30:(2s,4r)-1-((s)-2-(2-(4-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丁醇-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.合成中间体:2-氯-4-((1R,3R)-3-(5-乙炔基-1-氧代异丁醇-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物4-((1R,3R)-3-(5-溴-1-氧代异丁醇-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(189mg,0.4mmol)和三甲基硅乙炔(59mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=3:1),得到147mg中间体17-1,收率:88%。质谱(ES):m/z=419[M+H]+。
2.合成中间体:2-(4-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酸叔丁酯
将化合物2-氯-4-((1R,3R)-3-(5-乙炔基-1-氧代异丁醇-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(17-1)(419mg,1mmol)和2-(4-叠氮丁氧基)乙酸叔丁酯(229mg,1mmol)溶于5mL DMF后,加入CuI(38mg,0.2mmol)和三乙胺(0.1mL),室温搅拌反应4h。反应完成后加入水并用乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(PE:EA=3:1),得到558mg中间体17-2,收率:86%。质谱(ES):m/z=648[M+H]+。
3合成中间体:2-(4-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酸
将化合物2-(4-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酸叔丁酯(中间体17-2)(65mg,0.1mmol)溶于2mL二氯甲烷中,加入1mL三氟乙酸,室温搅拌2h反应完成,减压除去溶剂,未经进一步纯化,直接用于下一步反应。
4.合成最终产物30:(2s,4r)-1-((s)-2-(2-(4-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丁醇-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
将化合物2-(4-(4-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)-1H-1,2,3-三唑-1-基)丁氧基)乙酸(59mg,0.1mmol)和((2s,4r)-1-((s)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(44.5mg,0.1mmol)溶于2mL二氯甲烷中,DIPEA(38.7mg,0.3mmol),HATU(76.1mg,0.2mmol)后,室温搅拌2h,反应完成后加入水并用二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品采用板层析纯化(MeOH:DCM=10:1),得到56mg目标产物,收率:55%。质谱(ES):m/z=1018[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.75(s,1H),8.45(d,J=7.7Hz,1H),8.09(s,1H),7.97(d,J=7.8Hz,1H),7.91(d,J=8.7Hz,1H),7.77(d,J=8.0Hz,1H),7.47-7.28(m,6H),7.08(d,J=8.7Hz,1H),5.35(s,1H),5.21(s,1H),4.81(d,J=36.8Hz,3H),4.44(ddd,J=61.5,38.1,19.5Hz,6H),3.98-3.72(m,4H),3.52(t,J=6.1Hz,2H),2.30-2.22(m,1H),1.99(d,J=5.2Hz,5H),1.59(d,J=7.3Hz,2H),1.39(d,J=20.9Hz,6H),1.34-1.21(m,4H),1.15(d,J=12.8Hz,6H),0.95(s,9H).
31:(2s,4r)-1-((s)-2-(2-(3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)丙氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((s)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
得到59mg最终产物31,收率:58%。质谱(ES):m/z=1021[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.75(s,1H),8.45(d,J=7.7Hz,1H),8.09(s,1H),7.97(d,J=7.8Hz,1H),7.91(d,J=8.7Hz,1H),7.77(d,J=8.0Hz,1H),7.47-7.28(m,5H),7.08(d,J=8.7Hz,1H),5.35(s,1H),5.21(s,1H),4.81(d,J=36.8Hz,3H),4.44(ddd,J=61.5,38.1,19.5Hz,4H),3.98-3.72(m,4H),3.52(t,J=6.1Hz,2H),3.48(m,8H),2.30-2.22(m,1H),1.99(d,J=5.2Hz,5H),1.59(d,J=7.3Hz,2H),1.39(d,J=20.9Hz,6H),1.34-1.21(m,4H),1.15(d,J=12.8Hz,6H),0.95(s,9H).
32:(2s,4r)-1-((s)-2-(2-((1-(4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异偶姻-5-基)-3-丁炔-1-基)氮杂环丁烷-3-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
得到51mg最终产物32,收率:50%。质谱(ES):m/z=1002[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.51-8.44(m,1H),7.91(s,1H),7.63(d,J=9.9Hz,2H),7.54-7.33(m,6H),7.29(s,1H),7.11-7.03(m,1H),5.25-5.17(m,2H),4.95-4.86(m,1H),4.77(s,2H),4.54(s,3H),4.31(s,1H),3.98(s,3H),3.82-3.73(m,1H),3.65(m,2H),3.45(m,2H),2.58(m,2H),2.46(m,4H),2.32-2.21(m,2H),1.39(s,8H),1.24(s,3H),1.15(s,6H),0.96(s,9H).
33:N-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(6-(2-((S)-1-((2R,4R)-4-羟基-2-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-1-基)-3,3-二甲基-1-氧代丁烷-2-基)氨基)-2-氧代乙氧基)-1-己炔-1-基)-N-(甲基-d3)烟酰胺
得到59mg最终产物33,收率:60%。质谱(ES):m/z=981[M+H]+。1H NMR(400MHz,DMSO-d6)δ8.99(s,1H),8.47(d,J=7.3Hz,2H),7.82(dd,J=50.3,8.4Hz,2H),7.56-6.89(m,12H),5.29(s,1H),5.20(s,1H),4.90(d,J=7.3Hz,1H),4.45(dd,J=15.1,8.9Hz,2H),3.98-3.71(m,5H),3.57(t,J=26.1Hz,4H),2.46(m,1H),2.35-2.22(m,2H),1.68(dd,J=12.9,6.1Hz,6H),1.36(s,9H),0.95(s,12H).
34:(2S,4R)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)-3-丁炔氧基)丙胺基)-3,3-二甲基丁醇基)-4-羟基-N-(S)-1-(4-(4-甲基噻唑)苯基)乙基)甲酰胺
1.化合物3-(3-丁炔氧基)丙酸乙酯的合成
化合物3-丁炔-1-醇(1.00g,14.27mmol)、丙烯酸乙酯(1.71g,17.12mmol)10mL乙腈和DBU(1.09g,7.13mmol)依次加入反应瓶,氮气保护,70℃反应过夜。TCL确定反应终点,加入水和乙酸乙酯萃取3次,有机层合并用10%柠檬酸水溶液洗涤1次,饱和食盐水洗涤3次,无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到无色油状化合物3-(3-丁炔氧基)丙酸乙酯(826mg,4.85mmol),收率:34%。LC/MS(ESI+)calcd for C9H14O3(M+H+)m/z,171.1;found,171.2.
2.化合物3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)-3-丁炔氧基)氧基)丙酸乙酯的合成
化合物3-(3-丁炔氧基)丙酸乙酯(150mg,0.32mmol),4-((1r,3r)-3-(5-溴-1-氧代异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苄腈(108mg,0.64mmol),[1,1'-双(二苯基膦基)二茂铁]二氯化钯(46mg,0.06mmol),碘化亚铜(30mg,0.16mmol),三乙胺0.5mL和甲苯1.5mL依次加入反应器,氮气保护,110℃反应过夜。TLC确定反应结束,过滤,滤饼用二氯甲烷淋洗,旋干,硅胶柱层析纯化,得到类白色固体3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)-3-丁炔氧基)氧基)丙酸乙酯(56mg,0.10mmol),收率:31%。
LC/MS(ESI+)calcd for C32H35ClN2O5(M+H+)m/z,563.2;found,563.2.
3.化合物(3R,5S)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)-3-丁炔氧基)氧基)丙胺基)-3,3-二甲基丁醇)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-3-基乙酸的合成
将化合物3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)-3-丁炔氧基)氧基)丙酸乙酯(50mg,0.09mmol)加入2mL甲醇溶清,一水合氢氧化锂(15mg,0.36mmol)用0.5mL水溶清加入反应器,室温搅拌3h。TLC确定反应终点,反应液加入水,1N盐酸调节pH至2-3,乙酸乙酯萃取3次,无水硫酸钠干燥,旋干得到粗品。粗品加入2mL DMF和DIPEA(54mg,0.42mmol),冰浴加入HATU(64mg,0.17mmol)搅拌0.5h,加入(3R,5S)-1-((S)-2-氨基-3,3-二甲基丁酰基)-5-((((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基))氨基甲酰基)吡咯烷-3-乙酸酯(45mg,0.09mmol),室温搅拌反应2h,TLC确定反应终点,加入水和乙酸乙酯萃取3次,饱和食盐水洗涤3次,无水硫酸钠干燥,旋干pre-TLC分离得到类白色固体化合物(3R,5S)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)-3-丁炔氧基)氧基)丙胺基)-3,3-二甲基丁醇)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-3-基乙酸(59mg,0.06mmol)。收率:70%。
LC/MS(ESI+)calcd for C55H63ClN6O8S(M+H+)m/z,1003.4;found,1003.5.
1H NMR(400MHz,CDCl3)δ8.78(s,1H),7.76(d,J=7.8Hz,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=8.0Hz,1H),7.45(s,1H),7.43–7.31(m,5H),7.00(d,J=2.4Hz,1H),6.87–6.78(m,2H),5.35(s,1H),5.08(p,J=7.1Hz,1H),4.73(dd,J=8.2,6.3Hz,1H),4.63(s,2H),4.56(d,J=8.9Hz,1H),4.39(s,1H),4.32(s,1H),4.07(d,J=11.4Hz,1H),3.82–3.77(m,2H),3.72(t,J=7.1Hz,2H),2.79–2.70(m,3H),2.56(s,3H),2.53(t,J=6.1Hz,2H),2.16–2.07(m,2H),2.06(s,3H),1.48(d,J=6.9Hz,3H),1.43(d,J=12.3Hz,6H),1.26(s,6H),1.05(d,J=8.6Hz,9H).
4.化合物(2S,4R)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)-3-丁炔氧基)丙胺基)-3,3-二甲基丁醇基)-4-羟基-N-(S)-1-(4-(4-甲基噻唑)苯基)乙基)甲酰胺的合成
将化合物(3R,5S)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)-3-丁炔氧基)氧基)丙胺基)-3,3-二甲基丁醇)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)吡咯烷-3-基乙酸(24mg,0.02mmol)加入2mL甲醇溶清,一水合氢氧化锂(4mg,0.10mmol)用0.5mL水溶清加入反应器,室温搅拌3h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,旋干得到粗品。pre-TLC分离得到淡黄色固体化合物(2S,4R)-1-((S)-2-(3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)-3-丁炔氧基)丙胺基)-3,3-二甲基丁醇基)-4-羟基-N-(S)-1-(4-(4-甲基噻唑)苯基)乙基)甲酰胺(21mg,0.02mmol)。收率:91%。LC/MS(ESI+)calcd for C53H61ClN6O7S(M+H+)m/z,961.4;found,961.4.1H NMR(400MHz,CDCl3)δ8.69(s,1H),7.76(d,J=7.9Hz,1H),7.58(d,J=8.7Hz,1H),7.52–7.43(m,3H),7.39(q,J=8.4Hz,4H),6.99(t,J=6.3Hz,2H),6.84(dd,J=8.7,2.4Hz,1H),5.12–5.02(m,1H),4.75(t,J=7.9Hz,1H),4.63(s,2H),4.51(d,J=8.1Hz,2H),4.40(s,1H),4.32(s,1H),4.15(d,J=11.6Hz,1H),3.78(d,J=2.6Hz,2H),3.72(t,J=7.0Hz,2H),3.57(dd,J=11.4,3.3Hz,1H),2.76(t,J=7.0Hz,2H),2.53(d,J=7.3Hz,5H),2.12–2.02(m,1H),1.46(dd,J=10.7,3.9Hz,9H),1.25(s,9H),1.02(d,J=30.5Hz,9H).
35:(2S,4R)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺的合成
1.化合物4-(4-碘-1H-吡唑-1-基)丁酸甲酯的合成
化合物4-碘吡唑(1.00g,5.16mmol)、4-溴丁酸甲酯(1.03g,5.68mmol)50mL乙腈和碳酸钾(1.07g,7.73mmol)依次加入反应瓶,氮气保护,室温搅拌反应过夜。TCL确定反应终点,加入水和乙酸乙酯萃取3次,有机层合并用10%柠檬酸水溶液洗涤1次,饱和食盐水洗涤3次,无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到无色油状化合物4-(4-碘-1H-吡唑-1-基)丁酸甲酯(1.23mg,4.18mmol),收率:81%。
LC/MS(ESI+)calcd for C8H11IN2O2(M+H+)m/z,295.0;found,295.0.
2.化合物4-(4-((三甲基硅基)乙炔基)-1H-吡唑-1-基)丁酸甲酯的合成
化合物4-(4-碘-1H-吡唑-1-基)丁酸甲酯(1.00g,3.40mmol),三甲基硅炔(510mg,5.10mmol),[1,1'-双(二苯基膦基)二茂铁]二氯化钯(249mg,0.34mmol),碘化亚铜(259mg,1.36mmol),三乙胺6mL和甲苯17mL依次加入反应器,氮气保护,80℃反应过夜。TLC确定反应结束,过滤,滤饼用二氯甲烷淋洗,旋干,硅胶柱层析纯化,得到棕黄色油状物4-(4-((三甲基硅基)乙炔基)-1H-吡唑-1-基)丁酸甲酯(806mg,3.05mmol),收率:90%。
LC/MS(ESI+)calcd for C13H20N2O2Si(M+H+)m/z,265.1;found,265.1.
3.化合物4-(4-乙炔基-1H-吡唑-1-基)丁酸甲酯合成
化合物4-(4-((三甲基硅基)乙炔基)-1H-吡唑-1-基)丁酸甲酯(800mg,3.03mmol),TBAF(1.91g,7.30mmol)和20mL THF依次加入反应器,氮气保护,室温搅拌反应1h。TLC确定反应结束,反应液加入水,EA萃取3次合并,用饱和食盐水洗涤2次,合并干燥旋干,得到棕黄色油状物4-(4-乙炔基-1H-吡唑-1-基)丁酸甲酯(580mg,3.02mmol)粗品,粗收率:100%。LC/MS(ESI+)calcd for C10H12N2O2(M+H+)m/z,193.1;found,193.1.
3.化合物4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁酸甲酯的合成
化合物4-(4-乙炔基-1H-吡唑-1-基)丁酸甲酯(151mg,0.78mmol)粗品,4-((1r,3r)-3-(5-溴-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苄腈(200mg,0.42mmol),[1,1'-双(二苯基膦基)二茂铁]二氯化钯(59mg,0.081mmol),碘化亚铜(55mg,0.29mmol),三乙胺1mL和甲苯3mL依次加入反应器,氮气保护,110℃反应过夜。TLC确定反应结束,过滤,滤饼用二氯甲烷淋洗,旋干,硅胶柱层析纯化,得到淡黄色固体4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)乙炔基)-1H-吡唑-1-基)丁酸甲酯(228mg,0.10mmol),收率:92%。LC/MS(ESI+)calcd for C33H33ClN4O4(M+H+)m/z,545.2;found,545.2.
4.化合物(3R,5S)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁酰胺基)-3,3-二甲基丁醇基)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)-3-吡咯烷基乙酸酯的合成
将化合物4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁酸甲酯(130mg,0.22mmol)加入2mL甲醇溶清,一水合氢氧化锂(37mg,0.89mmol)用0.5mL水溶清加入反应器,室温搅拌3h。TLC确定反应终点,反应液加入水,1N盐酸调节pH至2-3,乙酸乙酯萃取3次,无水硫酸钠干燥,旋干得到粗品。粗品加入3mL DCM和DIPEA(135mg,1.04mmol),冰浴加入HATU(127mg,0.33mmol)搅拌0.5h,加入(3R,5S)-1-((S)-2-氨基-3,3-二甲基丁酰基)-5-((((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基))氨基甲酰基)吡咯烷-3-乙酸酯(151mg,0.29mmol),室温搅拌反应2h,TLC确定反应终点,加入水和乙酸乙酯萃取3次,无水硫酸钠干燥,旋干pre-TLC分离得到类白色固体化合物(3R,5S)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁酰胺基)-3,3-二甲基丁醇基)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)-3-吡咯烷基乙酸酯(123mg,0.12mmol)。收率:53%。LC/MS(ESI+)calcd for C57H63ClN8O7S(M+H+)m/z,1039.4;found,1039.5.1H NMR(400MHz,CDCl3)δ8.77(s,1H),7.80(d,J=7.8Hz,1H),7.67(t,J=8.6Hz,2H),7.56(t,J=7.7Hz,2H),7.52(s,1H),7.42–7.35(m,4H),7.33(d,J=7.8Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),6.56(d,J=8.6Hz,1H),5.07(p,J=7.0Hz,1H),4.74(dd,J=8.2,6.5Hz,1H),4.66(s,2H),4.54–4.49(m,1H),4.40(s,1H),4.33(s,1H),4.21(dt,J=14.3,7.1Hz,2H),4.10(d,J=11.6Hz,1H),3.81(dd,J=11.6,4.7Hz,1H),2.74(dt,J=13.7,6.0Hz,1H),2.55(s,3H),2.25–2.10(m,6H),2.06(s,3H),1.47(d,J=9.3Hz,9H),1.26(s,6H),1.06(d,J=6.0Hz,9H).
5.化合物(2S,4R)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺的合成
将化合物(3R,5S)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丁酰胺基)-3,3-二甲基丁醇基)-5-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)氨甲酰)-3-吡咯烷基乙酸酯(60mg,0.058mmol)加入2mL甲醇溶清,一水合氢氧化锂(24mg,0.58mmol)用0.5mL水溶清加入反应器,室温搅拌3h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,旋干得到粗品。prep-TLC分离得到类白色固体化合物(2S,4R)-1-((S)-2-(4-(4-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)乙炔基)-1H-吡唑-1-基)丁胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(50mg,0.050mmol)。收率:87%。
LC/MS(ESI+)calcd for C55H61ClN8O6S(M+H+)m/z,997.4;found,997.5.1H NMR(400MHz,CDCl3)δ8.73(s,1H),7.73(d,J=7.9Hz,1H),7.61(d,J=3.2Hz,2H),7.50(t,J=8.4Hz,2H),7.45(s,1H),7.41(d,J=7.7Hz,1H),7.36–7.28(m,4H),6.92(dd,J=8.2,5.2Hz,2H),6.77(dd,J=8.7,2.4Hz,1H),5.01(p,J=6.8Hz,1H),4.72(t,J=7.9Hz,1H),4.59(s,2H),4.44(dd,J=11.8,6.1Hz,2H),4.34(s,1H),4.26(s,1H),4.21–4.08(m,3H),3.52(dd,J=11.4,3.2Hz,1H),2.49(s,4H),2.11(ddd,J=24.9,14.0,5.4Hz,6H),1.40(d,J=7.9Hz,9H),1.18(d,J=7.8Hz,6H),0.99(d,J=20.9Hz,9H).
36:(2S,4R)-1-((S)-2-(2-((6-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯并[3,4-b]吡啶基-2-基)己基-5-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺的合成
LC/MS(ESI+)calcd for C53H62ClN7O7S(M+H+)m/z,108.4;found,996.4.
1H NMR(400MHz,CDCl3)δ8.77(s,1H),7.98(d,J=7.9Hz,1H),7.51(d,J=8.7Hz,1H),7.40(t,J=7.6Hz,2H),7.36–7.29(m,4H),7.13(d,J=8.6Hz,1H),6.93(t,J=2.7Hz,1H),6.76(dd,J=8.7,2.4Hz,1H),5.01(p,J=6.9Hz,1H),4.69(t,J=7.7Hz,1H),4.62(s,2H),4.47(d,J=8.7Hz,2H),4.38(s,1H),4.23(s,1H),3.89(q,J=15.4Hz,2H),3.57–3.48(m,3H),2.51(s,6H),1.98(s,3H),1.73(ddd,J=19.9,13.9,7.2Hz,4H),1.41(d,J=6.9Hz,3H),1.38(s,6H),1.19(d,J=1.3Hz,6H),1.00(s,9H).
37:(2S,4R)-1-((2S)-2-(2-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)丁-3-炔-2-基)氧基)乙氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺的合成
得到类白色固体化合物37(14mg,0.014mmol)。LC/MS(ESI+)calcd forC54H63ClN6O8S(M+H+)m/z,991.4;found,991.5.1H NMR(400MHz,CDCl3)δ8.78(s,1H),7.83–7.74(m,1H),7.57(d,J=8.6Hz,1H),7.52(d,J=8.1Hz,3H),7.38(s,5H),6.99(s,1H),6.83(d,J=7.4Hz,1H),5.07(s,1H),4.75(s,1H),4.64(s,2H),4.58–4.44(m,3H),4.40(s,1H),4.31(s,1H),4.10(s,1H),4.05(s,2H),3.99(s,1H),3.72(t,J=14.8Hz,5H),3.60(d,J=9.5Hz,1H),2.08(s,1H),1.56(d,J=3.7Hz,3H),1.46(d,J=12.9Hz,9H),1.25(s,9H),1.07(s,9H).
38:(2R,4R)-1-((S)-2-(2-((6-(2-((1r,3r)-3-)3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-3-氧代-2,3-二氢-1H-吡咯并[3,4-c]吡啶基-6-基)己-5-基-炔-1-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺的合成
LC/MS(ESI+)calcd for C53H62ClN7O7S(M+H+)m/z,996.4;found,996.7
1H NMR(400MHz,CDCl3)δ9.01(s,1H),8.70(s,1H),8.02(s,1H),7.58(d,J=8.7Hz,1H),7.48(s,2H),7.44–7.31(m,4H),7.23(s,2H),6.99(d,J=2.0Hz,1H),6.90–6.77(m,1H),5.16–5.00(m,1H),4.75(s,1H),4.68(s,2H),4.62–4.47(m,2H),4.41(s,1H),4.30(s,1H),4.10(d,J=11.2Hz,1H),3.96(q,J=15.2Hz,2H),3.61(d,J=14.3Hz,3H),2.54(d,J=8.4Hz,5H),1.79(dd,J=23.9,6.3Hz,4H),1.53–1.38(m,8H),1.25(s,6H),1.06(s,9H).
39:(2S,4R)-1-((S)-2-(2-(4-(3-(2((1s,3s)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)丙-2-炔-1-基)哌啶-1-基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺
LC/MS(ESI+)calcd for C56H66ClN7O6S(M+H+)m/z,1000.5;found,1000.5.
1H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.44(d,J=7.6Hz,1H),7.90(d,J=8.8Hz,1H),7.85(d,J=9.7Hz,1H),7.65(d,J=8.3Hz,3H),7.54(dd,J=7.9,1.4Hz,1H),7.43(d,J=8.2Hz,3H),7.29(d,J=2.4Hz,1H),7.06(dd,J=8.8,2.4Hz,1H),5.32(t,J=4.8Hz,1H),5.14(d,J=3.4Hz,1H),4.77(s,3H),4.53(d,J=5.0Hz,2H),4.47-4.43(m,2H),3.59(d,J=14.9Hz,4H),3.41-3.36(m,2H),3.02(d,J=16.4Hz,2H),2.89(t,J=17.4Hz,6H),2.70-2.62(m,1H),2.04(d,J=11.3Hz,2H),2.02-1.96(m,1H),1.76(d,J=12.7Hz,5H),1.15(s,9H),0.94(d,J=6.1Hz,12H).
40:(2R,4R)-1-((S)-2-(2-((1-(3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)丙-2-炔-1-基)氮杂环丁烷-3-基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺
41:(2S,4R)-1-((S)-2-(1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)六-5-yn-1-基)氧基)环丙烷甲酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
1.化合物1-(5-戊炔基-1-氧)环丙酸乙酯合成
氮气保护下,1-羟基环丙烷羧酸甲酯(100.0mg,0.86mmol)溶于2mL THF,冰浴,加入NaH(41.0mg,1.03mmol),搅拌30分钟,再加入6-碘代-1-炔(179.0mg,0.86mmol)。室温搅拌过夜。加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物1-(5-戊炔基-1-氧)环丙酸乙酯(110.0mg,0.56mmol)。收率65.1%。
LC/MS(ESI+)calcd for C11H17O3 +([M+H]+)m/z:197.1;found 197.1。
2.化合物1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)六-5-炔-1-基)氧基)环丙烷羧酸甲酯的合成
氮气保护下,1-(5-戊炔基-1-氧)环丙酸乙酯(110.0mg,0.56mmol),4-((1r,3r)-3-(5-溴-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(100.0mg,0.21mmol),Pd(PPh3)2Cl2(50.0mg,0.05mmol),CuI(4.0mg,0.1mmol),0.5mL三乙胺加入到2mL甲苯中。加热到110℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)六-5-炔-1-基)氧基)环丙烷羧酸甲酯(65.0mg,0.13mmol)。收率52.3%。
LC/MS(ESI+)calcd for C34H38ClN2O5 +([M+H]+)m/z:589.2;found 589.1。
3.化合物1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)十六-5-炔-1-基)氧基)环丙烷羧酸的合成
1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)六-5-炔-1-基)氧基)环丙烷羧酸甲酯(65.0mg,0.13mmol),溶于2mL甲醇,加入2mL的2N NaOH。室温搅拌2h。用1N HCl条件pH至4-5,加入二氯甲烷萃取,有机层用饱和食盐水洗涤,干燥,旋干。得到化合物1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)十六-5-炔-1-基)氧基)环丙烷羧酸(65.0mg,0.13mmol)。收率100.0%。
4.化合物(2S,4R)-1-((S)-2-(1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)六-5-yn-1-基)氧基)环丙烷甲酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺的合成
1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)十六-5-炔-1-基)氧基)环丙烷羧酸(50.0mg,0.08mmol),HATU(37.0mg,0.08mmol),DIEA(35.0mg,0.24mmol)溶于2mL DMF,加入(2R,4R)-1-((S)-2-氨基-3,3-二甲基丁醇)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(46.0mg,0.08mmol)。室温搅拌2h。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到(2S,4R)-1-((S)-2-(1-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)六-5-yn-1-基)氧基)环丙烷甲酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺(40.0mg,0.04mmol)。收率44.9%。LC/MS(ESI+)calcd for C56H66ClN6O7S+([M+H]+)m/z:1001.4;found1001.5.1H NMR(400MHz,CDCl3)δ8.75(s,1H),7.76(d,J=7.8Hz,1H),7.57(d,J=8.7Hz,1H),7.47(d,J=7.9Hz,2H),7.44(s,1H),7.39(q,J=8.3Hz,4H),7.31(d,J=8.4Hz,1H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),5.14–5.02(m,1H),4.81–4.73(m,1H),4.66–4.57(m,2H),4.54–4.45(m,2H),4.42–4.36(m,1H),4.35–4.28(m,1H),4.21–4.12(m,1H),3.63–3.52(m,3H),2.65(d,J=9.4Hz,1H),2.59–2.46(m,6H),2.14–2.04(m,2H),1.82–1.71(m,4H),1.47(d,J=6.9Hz,3H),1.43(d,J=5.8Hz,6H),1.25(s,6H),1.07(d,J=17.6Hz,11H),0.88(t,J=6.7Hz,2H).
42:((2S,4R)-1-((S)-2-(3-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)乙炔基)-1H-吡唑-1-基)丙酰胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
得到化合物42(45.0mg,0.04mmol)。LC/MS(ESI+)calcd for C54H60ClN8O6S+([M+H]+)m/z:983.4;found 983.4.1H NMR(400MHz,CDCl3)δ9.02(s,1H),7.79(d,J=7.8Hz,1H),7.67(d,J=15.9Hz,2H),7.58(d,J=8.7Hz,1H),7.53(d,J=8.6Hz,1H),7.50(s,1H),7.45(s,1H),7.37(d,J=10.7Hz,4H),7.00(d,J=2.2Hz,1H),6.88–6.81(m,1H),6.62(s,1H),5.06(d,J=6.1Hz,1H),4.78(s,1H),4.64(s,2H),4.39(s,1H),4.33(s,1H),4.10(d,J=12.7Hz,1H),3.75–3.57(m,4H),2.84(d,J=25.6Hz,2H),2.62(s,4H),2.03(d,J=7.1Hz,2H),1.47–1.40(m,9H),1.28(d,J=3.0Hz,6H),1.00(s,9H).
43:(2S,4R)-1-((S)-2-(3-(3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)丙-2-yn-1-基)壬苷-1-基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
得到43(18.0mg,0.02mmol)。LC/MS(ESI+)calcd for C54H63ClN7O6S+([M+H]+)m/z:972.4;found 972.5.1H NMR(400MHz,CDCl3)δ8.67(s,1H),7.90(s,1H),7.78(d,J=8.2Hz,1H),7.57(d,J=8.7Hz,1H),7.50(d,J=7.0Hz,2H),7.43–7.31(m,4H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),5.11–5.02(m,1H),4.77(t,J=7.9Hz,1H),4.65(s,2H),4.51(d,J=8.8Hz,2H),4.40(s,1H),4.32(s,1H),4.12(d,J=11.9Hz,1H),3.85(s,2H),3.59(d,J=8.5Hz,1H),3.47(s,2H),2.95(s,1H),2.73(s,2H),2.53(d,J=3.7Hz,4H),2.13(s,1H),1.74(s,3H),1.47(d,J=8.3Hz,3H),1.45(s,6H),1.26(s,6H),1.07(s,9H).
44:((2S,4R)-1-((S)-2-(3-(3-(3-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)prop-2-yn-1-基)azetidin-1-基)丙胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
得到化合物44(20mg,0.02mmol)。LC/MS(ESI+)calcd for C55H65ClN7O6S+([M+H]+)m/z:986.4;found 986.5.1H NMR(400MHz,CDCl3)δ8.67(s,1H),8.20(s,1H),7.80(d,J=7.8Hz,1H),7.74(d,J=7.8Hz,1H),7.58(d,J=8.7Hz,1H),7.54–7.46(m,2H),7.38(s,4H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),5.12–5.05(m,1H),4.81(d,J=8.2Hz,1H),4.65(s,2H),4.58(d,J=8.3Hz,1H),4.47(s,1H),4.40(s,1H),4.31(s,1H),4.19(s,2H),4.10(d,J=11.5Hz,1H),3.78(d,J=31.2Hz,2H),3.61(d,J=8.9Hz,1H),3.38(s,1H),3.21(d,J=42.9Hz,2H),2.76(s,2H),2.64(s,2H),2.52(s,3H),2.34(s,2H),1.48(d,J=6.9Hz,3H),1.45(s,6H),1.25(s,6H),1.08(s,9H).
45:2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)壬二酸-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物(2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚啉-2-基)环丁氧基)苯甲腈的合成
氮气保护下,4-((1r,3r)-3-(5-溴-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(500.0mg,1.06mmol),乙炔基三甲基硅烷(518.0mg,5.28mmol),Pd(PPh3)2Cl2(100.0mg,0.1mmol),CuI(60.0mg,0.2mmol),2mL三乙胺加入到6mL甲苯中。加热到100℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚-2-基)环丁氧基)苯甲腈(400mg,0.80mmol)。收率77.2%。LC/MS(ESI+)calcd for C28H32ClN2O2Si+([M+H]+)m/z:491.2;found 491.1。
2.化合物2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚-2-基)环丁氧基)苯甲腈(400.0mg,0.80mmol),TBAF(1.0g,1.60mmol)溶于10mLTHF。室温搅拌过夜。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(150mg,0.35mmol)。收率43.9%。
LC/MS(ESI+)calcd for C25H24ClN2O2 +([M+H]+)m/z:419.2;found 419.1。
3.化合物3-(4-碘-1H-吡唑-1-基)壬二酸叔丁酯的合成
4-碘代-1H-吡唑(400.0mg,1.40mmol),叔丁基3-碘代泽定-1-羧酸酯(274.0mg,1.41mmol),K2CO3(292.0mg,2.12mmol)溶于5mL DMF。加热到85℃,搅拌过夜。冷却至室温。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物3-(4-碘-1H-吡唑-1-基)壬二酸叔丁酯(380mg,1.08mmol)。收率77.0%。
4.化合物1-(壬二酸-3-基)-4-碘-1H-吡唑的合成
3-(4-碘-1H-吡唑-1-基)壬二酸叔丁酯(380.0mg,1.08mmol)溶于2mL二氯甲烷和2mL三氟乙酸。室温搅拌2h。浓缩,旋干,得到化合物1-(壬二酸-3-基)-4-碘-1H-吡唑(180mg,1.08mmol)。收率100.0%。
5.化合物2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)壬二酸-1-基)异吲哚啉-1,3-二酮的合成
1-(壬二酸-3-基)-4-碘-1H-吡唑(180.0mg,1.08mmol),2-(2,6-二氧哌啶-3-基)-5,6-二氟异吲哚-1,3-二酮(100.0mg,0.339mmol),DIEA(219.0mg,1.70mmol)溶于3mLDMSO。加热到130℃,搅拌3h。冷却至室温。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)壬二酸-1-基)异吲哚啉-1,3-二酮(150mg,0.26mmol)。收率84.0%。
LC/MS(ESI+)calcd for C19H16FIN5O4 +([M+H]+)m/z:524.0;found 524.0。
6.化合物2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)壬二酸-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
氮气保护下,2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)壬二酸-1-基)异吲哚啉-1,3-二酮(50.0mg,0.10mmol),2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(40.0mg,0.10mmol),Pd(PPh3)2Cl2(8.0mg,0.01mmol),CuI(10.0mg,0.02mmol),0.3mL三乙胺加入到1mL甲苯中。加热到100℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)壬二酸-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(15mg,0.80mmol)。收率19.2%。
LC/MS(ESI+)calcd for C44H38ClFN7O6 +([M+H]+)m/z:814.3;found 814.2.
1H NMR(400MHz,CDCl3)δ8.05–7.98(m,1H),7.81(d,J=7.9Hz,1H),7.74(s,1H),7.64(s,1H),7.60–7.49(m,3H),7.41(d,J=12.2Hz,1H),7.09(d,J=7.2Hz,1H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),5.11–4.98(m,1H),4.97–4.88(m,1H),4.66(s,2H),3.81–3.69(m,3H),3.23(td,J=9.8,5.1Hz,1H),2.99–2.85(m,5H),2.85–2.70(m,2H),2.16–2.08(m,1H).1.46(s,6H),1.27(s,6H).
46:(3R,5S)-1-((S)-2-(2-((5-((4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)苯基)氨基)戊基)氧基)乙酰胺基)-3,3-二甲基丁醇基)-5-((1-(4-甲基噻唑-5-基)苯基)环丙基)氨甲酰)吡咯烷-3-基乙酸酯
1H NMR(400MHz,DMSO-d6)δ8.95(s,1H),8.87(s,1H),7.90(d,J=8.8Hz,1H),7.63(d,J=8.7Hz,2H),7.47(d,J=9.1Hz,1H),7.32(d,J=9.4Hz,1H),7.21(d,J=2.3Hz,1H),7.00(dd,J=8.8,2.3Hz,1H),6.59-6.51(m,2H),6.17(s,1H),5.28(s,1H),4.31(s,1H),4.06-4.03(m,1H),3.94(s,2H),3.49(t,J=6.3Hz,2H),3.05(d,J=5.5Hz,2H),2.69(s,6H),2.43(s,2H),2.04-1.96(m,4H),1.65-1.50(m,4H),1.44(d,J=6.4Hz,2H),1.20(d,J=4.6Hz,6H),1.17(d,J=7.1Hz,2H),1.11(s,9H),1.02-0.80(m,12H).LC/MS(ESI+)calcdfor C55H68ClN7O8S([M+H]+)m/z:1022.70;found 511.8。
47:(2S,4R)-1-((S)-2-(2-((5-((4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)苯基)氨基)戊基)氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-(1-(4-甲基噻唑-5-基)苯基)环丙基)吡咯烷-2-甲酰胺
1H NMR(400MHz,DMSO-d6)δ8.94(s,1H),8.84(s,1H),7.90(d,J=8.8Hz,1H),7.66-7.60(m,2H),7.45(d,J=9.5Hz,1H),7.33(s,2H),7.20(d,J=2.3Hz,1H),7.00(dd,J=8.8,2.4Hz,1H),6.54(d,J=8.7Hz,2H),6.16(s,1H),5.17(s,1H),4.56(d,J=9.7Hz,1H),4.42(t,J=8.3Hz,1H),4.35(s,1H),4.31(s,1H),4.03(d,J=7.1Hz,2H),3.94(s,1H),3.63(d,J=11.2Hz,1H),3.50(t,J=6.6Hz,3H),3.17(s,2H),3.05(d,J=5.9Hz,2H),2.43(d,J=6.4Hz,3H),1.99(s,3H),1.59(d,J=6.1Hz,4H),1.44(s,2H),1.11(s,9H),0.94(s,12H).LC/MS(ESI+)calcd for C53H66ClN7O7S([M+H]+)m/z:980.66;found 980.3
48:(2S,4R)-1-((S)-2-(2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基))-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)己基-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-2-羟基-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺
49:(3R,5S)-1-((S)-2-(2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基))-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)己基-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-5-(((R)-2-羟基-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯
1.叔丁基2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)己基(5-炔-1-基)氧基)乙酸酯的合成
照前述实施例合成得到,收率42%。LC/MS(ESI+)calcdfor:C35H41ClN2O5(M+H+)m/z,605.2;found,605.2.
2. 2-((6-(2-(((1r,3r-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)己基-5-炔-1-基)氧基)乙酸的合成
照前述实施例合成得到,收率80%。LC/MS(ESI+)calcdfor:C31H33ClN2O5(M+H+)m/z,549.2;found,549.2.
3.(2S,4R)-1-((S)-2-(2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基))-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)己基-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((R)-2-羟基-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺的合成
照前述实施例合成得到,收率43%。LC/MS(ESI+)calcdfor:C54H63ClN6O8S(M+H+)m/z,991.4;found,991.4.1H NMR(400MHz,CDCl3)δ7.77(d,J=7.6Hz,1H),7.57(d,J=8.7Hz,1H),7.54-7.37(m,6H),7.20(m,2H),6.99(d,J=2.9Hz,1H),6.84(d,J=8.8Hz,1H),5.17(s,1H),4.64(s,2H),4.54(s,3H),4.39(s,1H),4.31(s,1H),3.97(s,3H),3.83(m,1H),3.73(m,2H),3.59(s,2H),2.65(s,2H),2.51(m,2H),2.25-2.15(m,2H),1.25(s,19H),1.07(s,9H).
4.(3R,5S)-1-((S)-2-(2-((6-(2-(((1r,3r)-3-(3-氯-4-氰基苯氧基))-2,2,4,4-四甲基环丁基)-1-氧代异吲哚-5-基)己基-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-5-(((R)-2-羟基-1-(4-(4-甲基噻唑-5-基)苯基)乙基)氨基甲酰基)吡咯烷-3-基乙酸酯的合成
照前述实施例合成得到,收率40%。LC/MS(ESI+)calcdfor:C56H65ClN6O9S(M+H+)m/z,1033.4;found,1033.4.1H NMR(400MHz,CDCl3)δ9.23(s,1H),7.78(d,J=7.9Hz,1H),7.57(d,J=8.6Hz,1H),7.54-7.38(m,6H),7.16(d,J=9.1Hz,1H),6.99(d,J=2.5Hz,1H),6.84(d,J=8.6Hz,1H),5.15(s,1H),4.68-4.51(m,4H),4.40(s,1H),4.31(s,1H),4.13(d,J=11.9Hz,1H),4.07-3.94(m,3H),3.89(dd,J=11.7,4.4Hz,1H),3.60(s,2H),2.69(s,3H),2.52(t,J=6.7Hz,3H),2.31-2.18(m,2H),2.05(d,J=2.3Hz,2H),1.25(d,J=2.4Hz,19H),1.05(s,9H).
51:2-氯-4-((1R,3R)-3-(5-((1'-(2-(2,6-二氧代哌啶-3-基)-6氟-1,3-二氧异吲哚啉-5-基)-[1,4'-双哌啶]-4-基)乙炔基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
LC/MS(ESI+)Calcd for C48H48ClFN6O6(M+H+)m/z,859.3;found 859.3。1H NMR(400MHz,CDCl3)δ7.78(d,J=7.9Hz,1H),7.58(d,J=8.7Hz,1H),7.48(dd,J=12.8,6.1Hz,3H),7.39(d,J=7.3Hz,1H),7.00(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.64(s,2H),4.40(s,1H),4.32(s,1H),3.74(d,J=11.6Hz,2H),3.01(s,2H),2.88(dd,J=23.1,12.0Hz,4H),2.81–2.69(m,4H),2.62(s,1H),2.13(d,J=5.0Hz,3H),2.05(d,J=8.0Hz,4H),1.45(s,6H),1.26(s,6H).
52:2-氯-4-((1R,3R)-3-(5-((1'-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-[1,4'-双哌啶]-4-基)乙炔基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
LC/MS(ESI+)Calcd for C48H49ClN6O6(M+H+)m/z,841.3;found 841.3。1H NMR(400MHz,CDCl3)δ8.20–7.96(m,1H),7.78(d,J=7.8Hz,1H),7.70(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.53–7.44(m,2H),7.28(s,1H),7.07(d,J=6.7Hz,1H),7.00(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.2Hz,1H),4.94(dd,J=12.0,5.3Hz,1H),4.64(s,2H),4.40(s,1H),4.33(s,1H),4.05(d,J=12.1Hz,2H),3.75(s,1H),3.10–2.98(m,5H),2.95–2.69(m,6H),2.26(s,2H),2.16(s,2H),1.98(s,3H),1.45(s,6H),1.26(s,6H).
53:2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)壬二酸-3-基)哌啶-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
LC/MS(ESI+)Calcd for C46H45ClFN6O6(M+H+)m/z,813.3;found 813.3。1H NMR(400MHz,CDCl3)δ8.18(s,1H),7.78(d,J=7.9Hz,1H),7.65(d,J=8.3Hz,1H),7.57(d,J=8.7Hz,1H),7.54–7.45(m,2H),7.00(d,J=2.4Hz,1H),6.89–6.76(m,2H),6.54(dd,J=8.3,2.0Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.64(s,2H),4.40(s,1H),4.32(s,1H),4.12(t,J=7.5Hz,2H),3.94(s,2H),3.42(s,1H),3.01(d,J=16.1Hz,1H),2.92–2.81(m,2H),2.80–2.71(m,4H),2.29(s,2H),2.02(s,2H),1.84(d,J=9.1Hz,2H),1.45(s,6H),1.26(s,7H).
54:2-氯-4-((1S,4r)-4-(2-((3S)-3-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)吡咯烷-1-基)-5-氧-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈
LC/MS(ESI+)Calcd for C42H44ClFN8O5(M+H+)m/z,795.3;found 795.3。1H NMR(400MHz,DMSO-d6)δ7.88(d,J=8.7Hz,1H),7.69(d,J=8.7Hz,1H),7.45(d,J=11.5Hz,1H),7.40(d,J=2.4Hz,1H),7.31(d,J=7.8Hz,1H),7.18–7.11(m,1H),6.49(d,J=8.7Hz,1H),5.33(s,1H),5.09(d,J=12.9Hz,1H),4.57(s,2H),4.45(s,1H),4.38(d,J=17.6Hz,1H),4.26(d,J=8.8Hz,2H),4.07(s,2H),3.68(s,1H),3.58(s,1H),3.49–3.37(m,3H),3.21(s,1H),3.09(s,3H),2.91(s,2H),2.68(s,4H),2.60(d,J=17.0Hz,3H),2.46–2.38(m,2H),2.34(s,1H),2.14(s,2H),2.00(d,J=7.5Hz,2H),1.80(s,3H),1.57(s,2H),1.24(s,2H).
55:2-氯-4-((1S,4r)-4-(2-((3S)-3-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)吡咯烷-1-基)-5-氧-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈
LC/MS(ESI+)Calcd for C42H44ClFN8O5(M+H+)m/z,795.3;found 795.3。1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),7.88(d,J=8.7Hz,1H),7.69(d,J=8.7Hz,1H),7.45(d,J=11.5Hz,1H),7.40(d,J=2.4Hz,1H),7.31(d,J=7.8Hz,1H),7.18–7.11(m,1H),6.49(d,J=8.7Hz,1H),5.33(s,1H),5.09(d,J=12.9Hz,1H),4.57(s,2H),4.45(s,1H),4.38(d,J=17.6Hz,1H),4.26(d,J=8.8Hz,2H),4.07(s,2H),3.60(s,1H),3.49–3.37(m,3H),3.21(s,1H),3.15(s,2H),2.91(s,2H),2.68(s,4H),2.60(d,J=17.0Hz,3H),2.46–2.38(m,2H),2.34(s,1H),2.14(s,2H),2.00(d,J=7.5Hz,2H),1.80(s,3H),1.57(s,2H),1.24(s,2H).
56:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)Calcd for C43H47ClN8O5(M+H+)m/z,791.3;found 791.3。
57:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)Calcd for C43H45ClN8O6(M+H+)m/z,805.3;found 805.3。
1H NMR(400MHz,CDCl3)δ8.07(s,1H),7.84(d,J=8.7Hz,1H),7.74(s,1H),7.58(d,J=8.7Hz,1H),7.33(s,1H),7.11(s,1H),7.03(d,J=2.3Hz,1H),6.88(dd,J=8.7,2.4Hz,1H),6.68(d,J=8.9Hz,1H),4.97(dd,J=12.0,5.1Hz,1H),4.51(s,2H),4.32(d,J=7.4Hz,2H),4.22(s,2H),3.91(s,1H),3.75(s,1H),3.56(s,1H),3.46(s,2H),2.97(d,J=17.2Hz,4H),2.91(s,1H),2.88–2.82(m,1H),2.78(d,J=16.0Hz,1H),2.61(s,3H),2.26(s,4H),2.16(d,J=7.2Hz,2H),1.98(d,J=43.1Hz,5H),1.72(dd,J=16.4,8.0Hz,4H).
58:N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-1,4-二氮杂环庚烷-1-基)甲基)哌啶-1-基)苯甲酰胺的合成
1.化合物4-(4-(羟甲基)哌啶-1-基)苯甲酸甲酯的合成
将对氟苯甲酸甲酯(1.0g,6.5mmol)溶于30mL DMSO中,加入4-羟甲基哌啶(2.2g,19.5mmol)和碳酸钾(2.7g,19.5mmol),升温至100℃反应过夜。次日监测对氟苯甲酸甲酯消耗完毕,冷却至室温后加入水与乙酸乙酯萃取,有机相用0.05M HCl溶液,饱和食盐水洗。无水硫酸钠干燥,浓缩得到4-(4-(羟甲基)哌啶-1-基)苯甲酸甲酯1.1g。收率:68.1%。MS(ESI)m/e249.1(M+H)+。
2.化合物4-(4-(羟甲基)哌啶-1-基)苯甲酸的合成
将4-(4-(羟甲基)哌啶-1-基)苯甲酸甲酯(1.0g,4.0mmol)溶于8mLMeOH/THF(1:1),加入5N NaOH 3mL。室温搅拌16h。加入水和DCM,萃走有机杂质,留水相用1N HCl缓慢调节pH至4-5,析出大量白色固体。过滤,干燥滤饼得到4-(4-(羟甲基)哌啶-1-基)苯甲酸510.0mg。收率:54.1%。MS(ESI)m/e235.1(M+H)+。
3.化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(羟基甲基)哌啶-1-基)苯甲酰胺
将4-((1R,4R)-4-氨基环己基)氧基)-2-氯苯甲腈(328.0mg,1.3mmol)溶于DCM,降温至0℃,加入HATU(475.1,1.3mmol)与DIEA(461.4mg,3.6mmol),之后加入4-(4-(羟甲基)哌啶-1-基)苯甲酸(280.0mg,1.2mmol),恢复至室温反应2h。加CH2Cl2和水萃取,有机层用食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(羟基甲基)哌啶-1-基)苯甲酰胺370.0mg,收率:66.4%。MS(ESI)m/e467.2(M+H)+。
4.化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-甲酰基哌啶-1-基)苯甲酰胺
将N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(羟基甲基)哌啶-1-基)苯甲酰胺(370mg,0.8mmol)溶于10mL DCM与1mL四氢呋喃混合溶剂中,加入Dess-Martin(373.2mg,0.9mmol),室温反应1h。垫硅藻土过滤,滤液用亚硫酸氢钠水溶液饱和食盐水洗,无水硫酸钠干燥DCM相,过滤浓缩。得到N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-甲酰基哌啶-1-基)苯甲酰胺310.2mg。收率:81.4%。MS(ESI)m/e465.2(M+H)+。
5.4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚-5-基)-1,4-二氮杂环庚烷-1-羧酸叔丁酯
将2-(2,6-二氧代哌啶-3-基)-5-氟异吲哚啉-1,3-二酮(200.0mg,0.7mmol),1,4-二氮杂环庚烷-1-甲酸叔丁酯(174.1mg,0.9mmol),溶于6mL DMSO,加入0.5mL DIEA,100℃反应1.5h。冷却至室温,加入水和乙酸乙酯萃取,饱和食盐水洗,干燥浓缩,柱层析得到4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚-5-基)-1,4-二氮杂环庚烷-1-羧酸叔丁酯185.3mg。收率:56.2%。MS(ESI)m/e401.2(M-56+H)+。
6.5-(1,4-二氮杂环庚-1-基)-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮三氟乙酸盐
将4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚-5-基)-1,4-二氮杂环庚烷-1-羧酸叔丁酯(185.0mg,0.4mmol),溶于2mL DCM,加入6mL TFA,室温反应1h。直接浓缩至干备用。产物191mg即为5-(1,4-二氮杂环庚-1-基)-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮三氟乙酸盐。收率:99.0%。MS(ESI)m/e356.2(M+H)+。
7.化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-1,4-二氮杂环庚烷-1-基)甲基)哌啶-1-基)苯甲酰胺的制备
将5-(1,4-二氮杂环庚-1-基)-2-(2,6-二氧代哌啶-3-基)异二氢吲哚-1,3-二酮三氟乙酸盐(121.2mg,0.3mmol),与N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-甲酰基哌啶-1-基)苯甲酰胺(100.0mg,0.2mmol),溶于8mL混合溶剂(DCM/MeOH=5/1)中。加入三乙酰氧基硼氢化钠(170.0mg,0.8mmol),室温反应4h。加入水淬灭反应,DCM萃取。干燥浓缩后柱层析,得到N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-1,4-二氮杂环庚烷-1-基)甲基)哌啶-1-基)苯甲酰胺101.5mg。收率:58.8%。MS(ESI)m/e805.3(M+H)+。1H NMR(400MHz,CDCl3)δ8.20(s,1H),7.67(dd,J=8.6,3.4Hz,3H),7.57(d,J=8.7Hz,1H),7.12(d,J=2.1Hz,1H),7.01(d,J=2.3Hz,1H),6.94–6.83(m,4H),5.91(d,J=7.8Hz,1H),4.96(dd,J=12.2,5.3Hz,1H),4.29(dd,J=12.2,8.4Hz,1H),4.05(dd,J=11.0,7.3Hz,1H),3.82(d,J=12.7Hz,2H),3.64(t,J=6.0Hz,4H),2.97–2.73(m,7H),2.65(s,2H),2.38(s,2H),2.18(dd,J=28.4,13.7Hz,6H),1.99(s,2H),1.83(s,2H),1.69(s,4H),1.46–1.37(m,2H).
59:2-氯-4-(((1r,4r)-4-(2-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-4-基)哌啶-4-基)氧基)哌啶-1-基)-5-氧代-5H-吡咯并[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)Calcd for C43H44ClN7O7(M+H+)m/z,806.3;found 806.3。
1H NMR(400MHz,CDCl3)δ8.08(s,1H),7.84(d,J=8.8Hz,1H),7.59(t,J=8.1Hz,2H),7.40(d,J=7.0Hz,1H),7.20(d,J=8.4Hz,1H),7.03(d,J=2.3Hz,1H),6.88(dd,J=8.8,2.3Hz,1H),6.70(d,J=8.8Hz,1H),4.99(dd,J=12.1,5.3Hz,1H),4.31(s,2H),4.22(s,2H),4.07(d,J=15.7Hz,2H),3.76(s,2H),3.66–3.56(m,2H),3.52–3.42(m,2H),3.20(d,J=8.3Hz,2H),2.94–2.71(m,3H),2.26(s,2H),2.14(d,J=7.8Hz,1H),2.04(s,4H),1.94(s,2H),1.88(s,2H),1.71(d,J=8.4Hz,5H).
60:2-氯-4-(((1r,4r)-4-(2-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)-5-氧代-5H-吡咯并[3,4-b]吡啶-6(7H)-基)环己基)氧基)苄腈的合成
1.4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧代-6,7-二氢-5H-吡咯并[3,4]-b]吡啶-2-基)哌啶-4-基)氧基)哌啶-1-甲酸叔丁酯
将2-氯-4-(((1R,4R)-4-(2-氯-5-氧代-5H-吡咯并[3,4-b]吡啶-6(7H)-基)环己基)氧基)苄腈(100.0mg,0.3mmol)与4-(哌啶-4-基氧基)哌啶-1-羧酸叔丁酯(212.1mg,0.8mmol)溶于3mL NMP,加入0.5mL DIEA后升温至100℃过夜。冷却至室温,加入水和EA萃取,有机相再用水,0.5M HCl水溶液,饱和食盐水洗。然后用无水硫酸钠干燥,浓缩。得到4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧代-6,7-二氢-5H-吡咯并[3,4]-b]吡啶-2-基)哌啶-4-基)氧基)哌啶-1-甲酸叔丁酯142.1mg。收率:88.2%。MS(ESI)m/e594.3(M-56+H)+。
2.2-氯-4-(((1R,4R)-4-(5-氧代-2-(4-(哌啶-4-基氧基)哌啶-1-基)-5,7-二氢-6H-吡咯并[3,4-b]吡啶-6-基)环己基)氧基)苄腈三氟乙酸盐
将化合物4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧代-6,7-二氢-5H-吡咯并[3,4]-b]吡啶-2-基)哌啶-4-基)氧基)哌啶-1-甲酸叔丁酯(140.0mg,0.2mmol)溶于1mL DCM,然后加入3mL TFA,室温搅拌1h,监测反应完毕。直接减压浓缩至粘稠,再用DCM带至形成固体。得到2-氯-4-(((1R,4R)-4-(5-氧代-2-(4-(哌啶-4-基氧基)哌啶-1-基)-5,7-二氢-6H-吡咯并[3,4-b]吡啶-6-基)环己基)氧基)苄腈三氟乙酸盐142.1mg。收率:99.2%。MS(ESI)m/e549.1(M+H)+。
3.2-氯-4-(((1r,4r)-4-(2-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯并[3,4-b]吡啶-6-基)环己基)氧基)苄腈
将中间体2-氯-4-(((1R,4R)-4-(5-氧代-2-(4-(哌啶-4-基氧基)哌啶-1-基)-5,7-二氢-6H-吡咯并[3,4-b]吡啶-6-基)环己基)氧基)苄腈三氟乙酸盐(100.0mg,0.2mmol)溶于2mL DMSO,加入DIEA(97.3mg,0.8mmol),搅拌均匀后加入2-(2,6-二氧代哌啶-3-基)-5-氟异吲哚啉-1,3-二酮(49.9mg,0.2mmol)。升温至120℃反应2h,监测反应完毕。冷却至室温后,加入水和乙酸乙酯萃取。用0.5M盐酸水溶液,饱和食盐水洗有机相,无水硫酸钠干燥后浓缩,柱层析得到2-氯-4-(((1r,4r)-4-(2-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯并[3,4-b]吡啶-6-基)环己基)氧基)苄腈61.2mg。收率:41.6%。MS(ESI)m/e805.3(M+H)+。1H NMR(400MHz,CDCl3)δ8.11(s,1H),7.84(d,J=8.8Hz,1H),7.70(d,J=8.5Hz,1H),7.58(d,J=8.7Hz,1H),7.31(d,J=2.1Hz,1H),7.08(dd,J=8.6,2.1Hz,1H),7.03(d,J=2.3Hz,1H),6.88(dd,J=8.7,2.2Hz,1H),6.70(d,J=8.9Hz,1H),4.96(dd,J=12.1,5.2Hz,1H),4.32(d,J=8.0Hz,2H),4.22(s,2H),4.12–4.02(m,2H),3.86–3.68(m,4H),3.52–3.40(m,2H),3.37–3.28(m,2H),2.96–2.71(m,3H),2.26(s,2H),2.20–2.10(m,1H),2.06–1.89(m,6H),1.72(d,J=8.3Hz,8H).
61:N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-2-基)哌啶-1-基)哒嗪-3-羧酰胺的合成
1.化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氧代哌啶-1-基)哒嗪-3-甲酰胺的合成:
将6-氯-N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(200.0mg,0.51mmol)与4-哌啶酮三氟乙酸盐(217.9mg,1.02mmol)溶于5mL NMP,加入0.5mL DIEA后升温至100℃过夜。冷却至室温,加入水和EA萃取,有机相再用水,饱和柠檬酸水溶液洗,饱和食盐水洗。然后用无水硫酸钠干燥,浓缩。柱纯化得到N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氧代哌啶-1-基)哒嗪-3-甲酰胺120.1mg。收率:51.7%。MS(ESI)m/e453.9(M+H)+。
2.7-(2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯
将2-(2,6-二氧杂哌啶-3-基)-5,6-二氟异吲哚啉-1,3-二酮(500mg,1.7mmol),2-叔丁氧羰基-2,7-二氮杂螺[3.5]壬烷(461.54mg,2.04mmol)溶于8mL DMSO,加入0.5mLDIEA,升温至130℃反应2h。监测反应完毕。冷却至室温,加入10mL水,10mL乙酸乙酯萃取,饱和柠檬酸水溶液洗,饱和食盐水洗。然后用无水硫酸钠干燥,浓缩。柱纯化得到7-(2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯750.6mg。收率:88.2%。MS(ESI)m/e445.2(M-56+H)+
3.2-(2,6-二氧杂哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬基-7-基)异吲哚啉-1,3-二酮三氟乙酸盐
将7-(2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(750mg,1.51mmol),溶于4mL DCM,8mL三氟乙酸的混合溶剂中,室温搅拌1h。直接蒸空浓缩,用二氯甲烷带至体系成黄色固体,称量得到2-(2,6-二氧杂哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬基-7-基)异吲哚啉-1,3-二酮三氟乙酸盐822.6mg。收率:99.5%。MS(ESI)m/e400.1(M+H)+
4.化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-2-基)哌啶-1-基)哒嗪-3-羧酰胺
将化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氧代哌啶-1-基)哒嗪-3-甲酰胺(50.0mg,0.11mmol),与2-(2,6-二氧杂哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬基-7-基)异吲哚啉-1,3-二酮三氟乙酸盐(62.3mg,0.12mmol)溶于2mL二氯甲烷与1mL甲醇的混合溶剂中,加入三乙酰氧基硼氢化钠(93.4mg,0.44mmol),室温反应过夜。加水淬灭,二氯甲烷萃取,饱和食盐水洗,干燥有机相,过滤浓缩后纯化得到N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-2-基)哌啶-1-基)哒嗪-3-羧酰胺23.3mg。MS(ESI)m/e837.3(M+H)+。1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.60(d,J=8.1Hz,1H),7.85(d,J=8.8Hz,1H),7.80(d,J=9.5Hz,1H),7.71(d,J=11.4Hz,1H),7.44(d,J=7.4Hz,1H),7.39(d,J=2.3Hz,1H),7.34(d,J=9.6Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.53(s,1H),4.21(d,J=13.0Hz,2H),3.85(d,J=8.3Hz,1H),3.24(s,1H),3.17(s,4H),2.99(s,4H),2.91–2.80(m,1H),2.59(d,J=18.4Hz,1H),2.35(d,J=18.7Hz,2H),2.10(d,J=10.0Hz,2H),2.03(s,1H),1.89(d,J=10.2Hz,2H),1.81(s,4H),1.73(d,J=11.0Hz,2H),1.64(d,J=12.8Hz,2H),1.51(d,J=12.3Hz,2H),1.28–1.11(m,3H).
62:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)哒嗪-3-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H44ClN9O6(M+H+)m/z,806.3;found 806.3。
1H NMR(400MHz,CDCl3)δ8.16(s,1H),8.00(d,J=9.5Hz,1H),7.86(d,J=8.1Hz,1H),7.68(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.00(d,J=2.2Hz,3H),6.85(dd,J=8.8,2.2Hz,1H),6.76(d,J=7.1Hz,1H).4.94(dd,J=12.1,5.3Hz,1H),4.52(s,2H),4.31(s,2H),4.02(s,2H),3.59(s,2H),3.51(s,2H),3.40(s,2H),3.02(s,3H),2.93–2.66(m,6H),2.17(d,J=4.1Hz,6H),2.02(d,J=14.6Hz,2H),1.54–1.42(m,4H).
63:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H44ClN9O6(M+H+)m/z,806.3;found 806.3。
1H NMR(400MHz,CDCl3)δ8.83(s,1H),8.19(s,1H),7.98(s,1H),7.70(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.40(d,J=8.1Hz,1H),7.05–6.97(m,2H),6.85(dd,J=8.8,2.3Hz,1H),6.78(d,J=7.4Hz,1H),4.94(dd,J=12.1,5.3Hz,1H),4.52(s,2H),4.31(s,2H),4.02(s,2H),3.59(s,2H),3.51(s,2H),3.40(s,2H),3.02(s,3H),2.93–2.66(m,6H),2.17(d,J=4.1Hz,6H),2.02(d,J=14.6Hz,2H),1.53–1.41(m,4H).
64:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H43ClFN9O6(M+H+)m/z,826.3;found 826.3。
1H NMR(400MHz,CDCl3)δ8.80(s,1H),8.21(d,J=9.5Hz,1H),7.86(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.17(d,J=6.7Hz,1H),7.00(dd,J=5.9,3.6Hz,2H),6.85(dd,J=8.7,2.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.49(s,2H),4.32(t,J=10.1Hz,2H),4.05(d,J=8.2Hz,2H),3.53(s,4H),3.12(s,4H),2.91–2.66(m,6H),2.20–2.10(m,7H),1.51–1.35(m,4H).
65:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)哒嗪-3-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H43ClFN9O6(M+H+)m/z,826.3;found 826.3。
1H NMR(400MHz,CDCl3)δ8.10(s,1H),8.00(d,J=9.5Hz,1H),7.86(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.45(d,J=11.8Hz,1H),7.17(d,J=6.7Hz,1H),7.00(dd,J=5.9,3.6Hz,2H),6.85(dd,J=8.7,2.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.49(s,2H),4.32(t,J=10.1Hz,2H),4.05(d,J=8.2Hz,2H),3.53(s,4H),3.14(s,4H),2.91–2.66(m,6H),2.20–2.10(m,7H),1.54–1.38(m,4H).
66:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-(5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异戊醇-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)嘧啶-5-甲酰胺的合成
LC/MS(ESI+)Calcd for C42H44ClN9O6(M+H+)m/z,806.3;found 806.3。
1H NMR(400MHz,CDCl3)δ8.81(s,1H),8.18(d,J=9.5Hz,1H),7.86(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.17(d,J=6.7Hz,1H),7.00(dd,J=5.9,3.6Hz,2H),6.85(dd,J=8.7,2.4Hz,1H),4.82(dd,J=12.3,5.3Hz,1H),4.56(s,2H),4.32(t,J=10.1Hz,2H),4.05(d,J=8.2Hz,2H),3.51(s,4H),3.12(s,4H),2.91–2.66(m,6H),2.20–2.10(m,7H),1.55–1.38(m,4H).
67:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哒嗪-3-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H43ClFN9O6(M+H+)m/z,824.3;found 824.3。
1H NMR(400MHz,CDCl3)δ7.99(d,J=9.4Hz,1H),7.91(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.45(d,J=11.0Hz,1H),7.37(d,J=7.3Hz,1H),7.00(d,J=2.4Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.74(d,J=9.4Hz,1H),4.92(dd,J=12.3,5.3Hz,3H),4.32(t,J=9.9Hz,1H),4.11–3.99(m,2H),3.84(s,2H),3.59(dd,J=24.3,11.8Hz,5H),3.08(s,2H),2.95–2.86(m,5H),2.83–2.62(m,6H),2.32(d,J=10.1Hz,4H),2.16(dd,J=22.5,11.8Hz,7H),2.05–1.95(m,4H),1.89(s,2H),1.71(dt,J=22.4,10.2Hz,5H),1.54–1.40(m,3H).
68:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)嘧啶-5-甲酰胺的合成
LC/MS(ESI+)Calcd for C42H44ClN9O6(M+H+)m/z,806.3;found 806.3。1H NMR(400MHz,CDCl3)δ8.67(s,2H),8.45(s,1H),7.66(d,J=8.4Hz,1H),7.55(d,J=8.7Hz,1H),7.02–6.95(m,2H),6.84(dd,J=8.8,2.4Hz,1H),6.75(s,1H),5.91(s,1H),4.94(s,1H),4.66(d,J=9.8Hz,2H),4.26(d,J=10.3Hz,1H),4.00(s,1H),3.64(d,J=7.5Hz,2H),3.37(d,J=7.2Hz,2H),3.08(s,3H),2.92–2.62(m,6H),2.14(s,5H),1.87(s,2H),1.46(s,5H),1.25(s,2H).
69:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H44ClN9O6(M+H+)m/z,806.3;found 806.3。
1H NMR(400MHz,CDCl3)δ8.81(d,J=1.3Hz,1H),7.72(d,J=1.3Hz,1H),7.56(d,J=8.7Hz,1H),7.45(d,J=11.1Hz,1H),7.38(dd,J=10.5,7.8Hz,2H),7.16(d,J=5.6Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.35–4.26(m,1H),4.08–3.98(m,1H),3.82(dd,J=11.0,8.1Hz,2H),3.62(d,J=12.3Hz,2H),3.50(d,J=11.0Hz,2H),3.05(s,2H),2.98–2.83(m,6H),2.80–2.71(m,2H),2.71–2.65(m,2H),2.33(d,J=10.3Hz,2H),2.21–2.11(m,6H),2.05(s,3H),1.77–1.61(m,5H),1.42(d,J=12.0Hz,3H).
70:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)Calcd for C42H43ClFN9O6(M+H+)m/z,824.3;found 824.3。
1H NMR(400MHz,CDCl3)δ8.85(d,J=1.3Hz,1H),7.72(d,J=1.3Hz,1H),7.56(d,J=8.7Hz,1H),7.45(d,J=11.1Hz,1H),7.38(dd,J=10.5,7.8Hz,2H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.35–4.26(m,1H),4.08–3.98(m,1H),3.82(dd,J=11.0,8.1Hz,2H),3.62(d,J=12.3Hz,2H),3.50(d,J=11.0Hz,2H),3.05(s,2H),2.98–2.83(m,6H),2.80–2.71(m,2H),2.71–2.65(m,2H),2.31(d,J=10.3Hz,2H),2.21–2.11(m,6H),2.05(s,3H),1.78–1.62(m,5H),1.46(d,J=12.0Hz,3H).
71:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺环[3.5]壬-7-基)哒嗪-3-甲酰胺的合成
LC/MS(ESI+)Calcd for C43H45ClFN9O6(M+H+)m/z,838.3;found 838.3。
1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),8.60(d,J=8.1Hz,1H),7.87(d,J=8.8Hz,1H),7.82(d,J=9.5Hz,1H),7.70(d,J=11.4Hz,1H),7.41(d,J=7.4Hz,1H),7.36(d,J=2.3Hz,1H),7.32(d,J=9.6Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.09(dd,J=12.8,5.4Hz,1H),4.53(s,1H),4.21(d,J=13.0Hz,2H),3.85(d,J=8.3Hz,1H),3.24(s,1H),3.17(s,4H),2.99(s,4H),2.91–2.80(m,1H),2.59(d,J=18.4Hz,1H),2.35(d,J=18.7Hz,2H),2.10(d,J=10.0Hz,2H),2.03(s,1H),1.89(d,J=10.2Hz,2H),1.81(s,4H),1.73(d,J=11.0Hz,2H),1.64(d,J=12.8Hz,2H),1.51(d,J=12.3Hz,2H),1.2–1.10(m,3H).
72:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(5-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哒嗪-3-甲酰胺
LC/MS(ESI+)Calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found 852.3。
1H NMR(400MHz,CDCl3)δ8.19(d,J=9.1Hz,1H),7.99(d,J=9.4Hz,1H),7.57(d,J=8.7Hz,1H),7.45(d,J=11.0Hz,1H),7.37(d,J=7.3Hz,1H),6.97(d,J=2.4Hz,1H),6.81(dd,J=8.7,2.4Hz,1H),6.76(d,J=9.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.55(s,1H),4.19(d,J=9.1Hz,1H),4.07(s,1H),3.85(s,2H),3.61(t,J=13.4Hz,4H),3.10(s,2H),2.91(dd,J=22.3,8.6Hz,5H),2.85–2.65(m,4H),2.13(dd,J=11.3,6.2Hz,2H),2.07–1.95(m,4H),1.74(dd,J=20.5,9.9Hz,2H),1.28(s,5H),1.21(s,6H).
73:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1-(2,6-二氧哌啶-3-基-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)吡嗪-2-甲酰胺的合成
1.叔丁基(1-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)哌啶-4-基)氨基甲酸酯的合成
将5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(400mg,1.02mmol)溶于10mL NMP,加入叔丁基哌啶-4-基氨基甲酸酯(246mg,1.23mmol),DIEA(661mg,5.11mmol),升温至100℃反应。加水淬灭,EA萃取,饱和柠檬酸水溶液洗。纯化后得到叔丁基(1-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)哌啶-4-基)氨基甲酸酯450mg。收率:79.3%。LC/MS(ESI+)Calcd for C28H35ClN6O4(M-56+H+)m/z,500.2;found 500.2。
2.5-(4-氨基哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)己基)嗪-2-甲酰胺三氟乙酸盐
将叔丁基(1-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)哌啶-4-基)氨基甲酸酯(300mg,0.54mmol),溶于2mL DCM,加入4mL三氟乙酸,搅拌2h。直接浓缩至干,用DCM带走大部分TFA。得到5-(4-氨基哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)己基)嗪-2-甲酰胺三氟乙酸盐325mg。
3.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1-(2,6-二氧哌啶-3-基-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)吡嗪-2-甲酰胺的合成
将5-(4-氨基哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)己基)嗪-2-甲酰胺三氟乙酸盐(100mg,0.22mmol),2-(2,6-二氧哌啶-3-基)-5-氟-6-(4-氧哌啶-1-基)异吲哚啉-1,3-二酮(83mg,0.22mmol),溶于2mL DCM和2mL甲醇的混合溶剂里。缓慢加入三乙酰氧基硼氢化钠(186mg,0.88mmol),反应5h。通过硅胶板纯化得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1-(2,6-二氧哌啶-3-基-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)吡嗪-2-甲酰胺12mg,收率:7.3%。LC/MS(ESI+)Calcd forC28H35ClN6O4(M+H+)m/z,812.3;found 812.3。1H NMR(400MHz,CDCl3)δ8.84(s,1H),7.99(s,1H),7.56(d,J=8.7Hz,1H),7.47(dd,J=11.0,2.7Hz,1H),7.40(dd,J=13.9,7.5Hz,2H),7.00(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.99–4.88(m,2H),4.48(s,2H),4.31(s,2H),4.05(s,2H),3.71(d,J=7.5Hz,2H),3.61–3.51(m,1H),3.16(s,1H),3.08(d,J=10.3Hz,3H),2.93(t,J=12.9Hz,4H),2.79(dd,J=25.3,13.3Hz,3H),2.18(d,J=4.6Hz,8H),2.07(s,1H),1.25(s,3H).
74:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺环[3.5]壬-7-基)哒嗪-3-甲酰胺的合成
LC/MS(ESI+)Calcd for C43H46ClN9O6(M+H+)m/z,820.3;found 820.3。
1H NMR(400MHz,CDCl3)δ8.21(s,1H),7.94(d,J=9.5Hz,1H),7.87(d,J=8.2Hz,1H),7.66(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.2Hz,1H),7.06(dd,J=8.6,2.2Hz,1H),7.00(t,J=5.7Hz,2H),6.86(dd,J=8.8,2.4Hz,1H),4.97(dd,J=12.3,5.3Hz,1H),4.55(s,2H),4.32(t,J=10.0Hz,1H),4.05(dd,J=11.4,7.2Hz,1H),3.43–3.37(m,4H),3.22(d,J=10.9Hz,3H),2.94–2.67(m,4H),2.24–2.10(m,6H),1.94(s,6H),1.78–1.63(m,8H),1.52–1.40(m,3H).
75:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮螺环[3.5]壬-7-基)哒嗪-3-甲酰胺的合成路线:
LC/MS(ESI+)Calcdfor C44H47ClFN9O6(M+H+)m/z,852.3;found 852.3。1H NMR(400MHz,CDCl3)δ8.46(d,J=2.3Hz,1H),7.58(dd,J=9.0,2.4Hz,1H),7.55(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.38(d,J=7.3Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),6.64(d,J=9.0Hz,1H),5.90(d,J=7.7Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.52–4.25(m,4H),4.04(d,J=7.7Hz,1H),3.33(s,3H),3.18(s,3H),3.04–2.65(m,6H),2.57(s,1H),2.27(s,4H),2.17(dd,J=25.9,12.6Hz,9H),2.05(s,3H),1.98(s,4H),1.86(d,J=11.4Hz,2H),1.76(d,J=12.9Hz,2H),1.48–1.36(m,4H).
76:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮螺环[3.5]壬-7-基)哒嗪-2-甲酰胺
LC/MS(ESI+)Calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found 852.3。1H NMR(400MHz,CDCl3)δ8.84(d,J=2.3Hz,1H),8.25(dd,J=9.0,2.4Hz,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.38(d,J=7.3Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),6.77(d,J=9.0Hz,1H),5.93(d,J=7.7Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.52–4.25(m,4H),4.04(d,J=7.7Hz,1H),3.33(s,3H),3.18(s,3H),3.04–2.65(m,6H),2.57(s,1H),2.27(s,4H),2.17(dd,J=25.9,12.6Hz,9H),2.05(s,3H),1.97(s,4H),1.85(d,J=11.4Hz,2H),1.67(d,J=12.9Hz,2H),1.45–1.33(m,4H).
77:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-2-基)哌啶-1-基)烟酰胺
LC/MS(ESI+)Calcd for C44H46ClFN8O6(M+H+)m/z,837.3;found 837.3。1H NMR(400MHz,CDCl3)δ8.84(d,J=2.3Hz,1H),8.25(dd,J=9.0,2.4Hz,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.38(d,J=7.3Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),6.77(d,J=9.0Hz,1H),5.93(d,J=7.7Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.52–4.25(m,4H),4.04(d,J=7.7Hz,1H),3.33(s,3H),3.18(s,3H),3.04–2.65(m,6H),2.57(s,1H),2.27(s,4H),2.17(dd,J=25.9,12.6Hz,9H),2.05(s,3H),1.97(s,4H),1.85(d,J=11.4Hz,2H),1.67(d,J=12.9Hz,2H),1.45–1.33(m,4H).
78:HC-4304-01:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-2-基)啶-1-基)嗪-3-甲酰胺
LC/MS(ESI+)Calcd forC43H46ClN9O6(M+H+)m/z,820.3;found 820.3。1H NMR(400MHz,CDCl3)δ8.21(s,1H),7.99(d,J=9.5Hz,1H),7.87(d,J=8.2Hz,1H),7.69(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.2Hz,1H),7.06(dd,J=8.6,2.2Hz,1H),7.00(t,J=5.7Hz,2H),6.86(dd,J=8.8,2.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.39(s,2H),4.32(t,J=10.0Hz,1H),4.05(dd,J=11.4,7.2Hz,1H),3.43–3.37(m,4H),3.22(d,J=10.9Hz,3H),2.94–2.67(m,4H),2.24–2.10(m,6H),1.94(s,6H),1.78–1.63(m,8H),1.52–1.40(m,3H).
79:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-2-基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)Calcd for C44H45F4N9O6(M+H+)m/z,872.3;found 872.3。1H NMR(400MHz,CDCl3)δ8.45(s,1H),8.00(d,J=9.5Hz,1H),7.89(d,J=8.2Hz,1H),7.75(d,J=8.6Hz,1H),7.47(d,J=10.8Hz,1H),7.38(d,J=7.2Hz,1H),7.26(s,1H),7.11(dd,J=8.7,2.4Hz,1H),7.00(d,J=9.6Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.48(d,J=13.3Hz,2H),4.40(dd,J=11.8,8.1Hz,2H),4.13–4.01(m,2H),3.49(s,4H),3.16(d,J=14.6Hz,7H),2.96–2.68(m,6H),2.24–2.16(m,7H),2.05(s,2H),2.02(s,6H),1.95(s,2H),1.77–1.67(m,3H),1.49(d,J=12.2Hz,3H).
80:N-((1s,3s)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-2-基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)Calcd for C45H49ClF4N9O6(M+H+)m/z,866.3;found 866.3。1H NMR(400MHz,CDCl3)δ8.55–8.41(m,1H),8.16(d,J=9.1Hz,1H),7.99(d,J=9.5Hz,1H),7.57(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.38(d,J=7.2Hz,1H),6.99(dd,J=12.0,6.0Hz,2H),6.81(dd,J=8.7,2.4Hz,1H),4.93(dd,J=12.3,5.3Hz,2H),4.44(d,J=13.4Hz,3H),4.19(d,J=9.0Hz,1H),4.07(s,1H),3.30(s,4H),3.18(d,J=6.2Hz,6H),2.96–2.72(m,6H),2.17–2.10(m,2H),2.05(s,4H),2.00–1.85(m,9H),1.51(d,J=10.6Hz,3H),1.28(s,6H),1.25(s,1H),1.21(s,6H).
81:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,6-二氮杂螺环[3.4]辛烷-2-基)哒嗪-3-甲酰胺
LC/MS(ESI+)Calcd for C41H43ClFN9O6s(M+H+)m/z,824.3;found 824.3。1H NMR(400MHz,CDCl3)δ8.20(s,1H),7.99(d,J=9.2Hz,1H),7.88(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.39(d,J=7.3Hz,1H),7.00(d,J=2.3Hz,1H),6.86(dd,J=8.7,2.3Hz,1H),6.61(d,J=9.2Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.32(t,J=9.8Hz,1H),4.19(dd,J=24.2,8.6Hz,4H),4.05(d,J=7.9Hz,1H),3.67(d,J=11.9Hz,2H),3.06(s,2H),2.98–2.70(m,8H),2.54–2.36(m,1H),2.27(s,2H),2.16(d,J=10.3Hz,6H),2.05(d,J=11.8Hz,3H),1.83(s,3H),1.69(d,J=11.8Hz,9H),1.47(dd,J=22.4,10.4Hz,3H).
82:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(6-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,6-二氮杂螺[3.4]辛烷-2-基)吡嗪-2-甲酰胺
LC/MS(ESI+)Calcd for C41H43ClFN9O6s(M+H+)m/z,824.3;found 824.3。1H NMR(400MHz,CDCl3)δ8.83(d,J=1.3Hz,1H),8.16(s,1H),7.62–7.53(m,2H),7.47(d,J=10.9Hz,1H),7.40(dd,J=12.6,7.8Hz,2H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.31(s,1H),4.20(s,2H),4.14(d,J=8.6Hz,2H),4.03(d,J=7.8Hz,1H),3.68(d,J=12.7Hz,2H),3.07(s,2H),2.91(dd,J=21.2,10.1Hz,4H),2.86–2.71(m,3H),2.62(s,2H),2.29(s,2H),2.24–2.11(m,6H),2.05(s,2H),1.88(s,2H),1.69(dd,J=22.2,9.9Hz,8H),1.53–1.39(m,3H).
83:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺
LC/MS(ESI+)Calcd for C41H43ClFN9O6s(M+H+)m/z,844.3;found 844.3。1H NMR(400MHz,CDCl3)δ8.00(s,1H),7.56(d,J=8.7Hz,1H),7.38(dd,J=11.6,5.3Hz,1H),7.05–6.97(m,2H),6.88–6.78(m,2H),4.91(dd,J=12.0,5.4Hz,1H),4.31(s,1H),4.13(s,2H),3.98(s,1H),3.95–3.85(m,4H),3.59(d,J=7.0Hz,2H),3.23(t,J=12.3Hz,2H),2.80(ddd,J=30.6,27.4,9.4Hz,6H),2.16(d,J=10.4Hz,6H),1.68(s,2H),1.62(s,4H),1.53–1.46(m,2H),1.25(s,6H).
84:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-(1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-3-羧酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-1,2,4-三唑-3-甲酰胺的合成
将1H-1,2,4-三氮唑-3-羧酸(500mg,1.99mmol)与10mL DMF混合,难溶。加入DIEA(309mg,2.39mmol),溶清。降温至-5℃,加入HATU(796mg,2.09mmol),搅拌1h以上,再加入4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苄腈(237mg,2.09mmol),低温搅拌30min,撤去冰盐浴,自然升温至室温,反应2h。搅拌下向反应体系中加水,析出大量固体。过滤,滤饼干燥得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-1,2,4-三唑-3-甲酰胺660mg。收率:95.7%。LC/MS(ESI+)Calcd for C16H16ClN5O2(M+H+)m/z,346.1;found 346.1。
2.(1R,4r)-甲基4-(3-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-1,2,4-三唑-1-基)环己酸盐的合成
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-1,2,4-三唑-3-甲酰胺(500mg,1.45mmol)溶于DMF,然后加入(1s,4s)-甲基4-((甲磺酰基)氧基)环己羧酸酯(1.02g,4.34mmol),和碳酸铯(1.41g,4.34mmol)。升温至95℃过夜反应。向体系中加入水淬灭,EA萃取,干燥,浓缩纯化得到(1R,4r)-甲基4-(3-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-1,2,4-三唑-1-基)环己酸盐210mg。收率:31.65%。LC/MS(ESI+)Calcdfor C24H28ClN5O4(M+H+)m/z,486.2;found 486.2。
3.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-1,2,4-三唑-3-甲酰胺
将(1R,4r)-甲基4-(3-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-1,2,4-三唑-1-基)环己酸盐(200mg,0.41mmol)溶于THF与甲醇1:1的混合溶剂中,缓慢加入硼氢化钠(62mg,1.65mmol),65℃回流反应2h,反应完全。冷却至室温,加水淬灭,EA萃取,干燥后通过硅胶板纯化得到N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-1,2,4-三唑-3-甲酰胺140mg。收率61.87%。LC/MS(ESI+)Calcd forC23H28ClN5O3(M+H+)m/z,458.2;found 458.2。
4.化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-1,2,4-三唑-3-甲酰胺的合成
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-1,2,4-三唑-3-甲酰胺(130mg,0.28mmol)溶于3mL DCM,室温下加入Dess-Martin试剂(181mg,0.43mmol),反应1h。过滤,滤液用亚硫酸氢钠洗,再用饱和碳酸氢钠溶液洗,干燥过滤。浓缩纯化得到N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-1,2,4-三唑-3-甲酰胺80mg。收率:61.84%5.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-(1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-3-羧酰胺
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-1,2,4-三唑-3-甲酰胺(40mg,0.09mmol),2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚林-1,3-二酮(37mg,0.10mmol)溶于2mL DCM:MeOH=1:1的混合溶剂,缓慢加入三乙酰氧基硼氢化钠(75mg,0.35mmol)。2h监测反应完毕。加水淬灭,DCM萃取。干燥纯化得到N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-(1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-3-羧酰胺35mg。收率:48.57%。LC/MS(ESI+)Calcd for C43H48ClN9O6(M+H+)m/z,822.3;found 822.3。1H NMR(400MHz,CDCl3)δ8.11(s,1H),7.64(d,J=8.3Hz,1H),7.55(d,J=8.7Hz,1H),7.15(d,J=8.2Hz,1H),6.98(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),6.77(d,J=1.7Hz,1H),6.50(dd,J=8.3,1.9Hz,1H),4.94(dd,J=12.3,5.3Hz,2H),4.72(d,J=25.2Hz,6H),4.25(dt,J=15.4,11.1Hz,3H),4.09(d,J=8.1Hz,1H),3.73(s,4H),2.81(ddd,J=33.0,24.0,10.4Hz,3H),2.40(s,3H),2.23–2.16(m,6H),1.87(s,6H),1.67(dd,J=23.0,10.0Hz,4H),1.47(dd,J=23.1,10.5Hz,2H),1.25(s,1H).
85:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-3-甲酰胺:
LC/MS(ESI+)Calcd for C41H43ClFN9O6s(M+H+)m/z,844.3;found 844.3。1H NMR(400MHz,CDCl3)δ8.63(s,1H),8.12(s,1H),7.56(d,J=8.7Hz,1H),7.36(d,J=10.8Hz,1H),7.18(d,J=8.3Hz,1H),6.99(d,J=2.4Hz,1H),6.87–6.77(m,2H),4.92(dd,J=12.2,5.3Hz,1H),4.63(s,2H),4.32–4.18(m,2H),4.15–4.05(m,1H),3.89(s,4H),2.97–2.65(m,4H),2.41(s,5H),2.27(d,J=6.9Hz,4H),2.15(dd,J=8.9,6.4Hz,5H),2.08(s,4H),1.92(s,4H),1.67(dd,J=22.6,9.8Hz,4H),1.47(d,J=12.7Hz,2H).
86:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-5-甲酰胺:
LC/MS(ESI+)Calcd for C43H48ClN9O6(M+H+)m/z,822.3;found 822.3。1H NMR(400MHz,CDCl3)δ8.35(s,1H),7.86(s,1H),7.64(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=8.2Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),6.77(d,J=2.0Hz,1H),6.50(dd,J=8.4,2.0Hz,1H),5.39(s,1H),4.93(dd,J=12.3,5.3Hz,2H),4.79(s,4H),4.30(t,J=10.2Hz,1H),3.99(d,J=8.3Hz,1H),3.73(s,4H),2.93–2.69(m,3H),2.28(s,4H),2.07(d,J=13.4Hz,4H),1.98(d,J=11.0Hz,4H),1.91–1.82(m,6H),1.70–1.60(m,3H),1.57–1.47(m,2H).
87:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-1,2,4-三唑-5-甲酰胺:LC/MS(ESI+)Calcd for C41H43ClFN9O6s(M+H+)m/z,844.3;found 844.3。1H NMR(400MHz,CDCl3)δ8.36(s,1H),7.86(s,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=8.2Hz,1H),7.36(d,J=10.9Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),6.80(d,J=7.5Hz,1H),5.39(t,J=11.5Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.58(s,4H),4.30(t,J=10.1Hz,1H),3.99(dd,J=11.6,7.4Hz,1H),3.89(d,J=1.7Hz,4H),2.95–2.65(m,4H),2.20(d,J=5.1Hz,4H),2.08(s,3H),2.04(d,J=4.3Hz,1H),1.97(t,J=11.2Hz,4H),1.89(s,4H),1.72–1.59(m,4H),1.51(dd,J=21.8,11.3Hz,3H).
88:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)哒嗪-3-甲酰胺的合成
1、4-(2-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯的合成
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2,7-二氮杂螺[3.5]壬-2-基)哒嗪-3-甲酰胺(100mg,0.208mmol)5mL二氯甲烷,加入4-Boc哌啶酮(414mg,2.1mmol),加入0.5滴乙酸,40℃回流1h,冷却至室温,加入三乙酰基硼氢化钠(441mg,2.08mmol),40℃回流过夜,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到产品4-(2-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯120mg,收率:86.8%,LC/MS(ESI+)calcd for C35H46ClN7O4([M+H]+)m/e663.8。
2、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异戊醇-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)哒嗪-3-甲酰胺的合成
将4-(2-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯(45mg,0.068mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,加入二氯甲烷再减压浓缩2次,加入3mL DMSO,加入DIEA0.5mL,加入2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮,135℃回流1.5h,冷却至室温,加入10mL二氯甲烷,10mL水洗涤2次,再用饱和食盐水洗涤1次,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)哒嗪-3-甲酰胺35mg,收率:62.7%,LC/MS(ESI+)calcd forC43H46ClN9O6([M+H]+)m/e 820.3,1H NMR(400MHz,CDCl3)δ8.20(s,1H),7.98(d,J=9.2Hz,1H),7.88(d,J=8.3Hz,1H),7.69(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.29(d,J=2.2Hz,1H),7.06(dd,J=8.6,2.2Hz,1H),7.00(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),6.58(d,J=9.3Hz,1H),4.95(dd,J=12.3,5.3Hz,1H),4.32(dd,J=11.7,8.1Hz,1H),4.03(t,J=11.6Hz,3H),3.91(s,4H),3.00(t,J=11.8Hz,2H),2.94–2.70(m,4H),2.62(s,4H),2.15(dt,J=10.4,7.8Hz,6H),1.97(s,4H),1.72–1.65(m,5H),1.53–1.43(m,2H).
89:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)吡嗪-2-羧酰胺的合成
合成步骤参照88
LC/MS(ESI+)calcd for C43H46ClN9O6([M+H]+)m/e 820.3,1H NMR(400MHz,CDCl3)δ8.83(d,J=1.2Hz,1H),8.09(s,1H),7.70(d,J=8.5Hz,1H),7.63–7.51(m,2H),7.39(d,J=8.2Hz,1H),7.29(d,J=2.0Hz,1H),7.07(dd,J=8.5,2.0Hz,1H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),4.99–4.90(m,1H),4.30(td,J=10.1,5.2Hz,1H),4.03(d,J=9.1Hz,3H),3.91(s,4H),3.00(t,J=12.1Hz,2H),2.94–2.72(m,4H),2.66(d,J=33.9Hz,4H),2.29–2.09(m,6H),1.99(s,4H),1.71(dd,J=18.7,8.9Hz,5H),1.48(dd,J=18.3,6.9Hz,2H).
90:2-氯-4-((1r,3r)-3-(2-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将2-氯-4-(3-(2-(4-甲酰哌啶-1-基)-5-氧代异吲哚啉-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(66mg,0.13mmol)溶于5mL二氯甲烷,加入2-(2,6-二氧异吲哚啉-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异苄-1,3-二酮(45mg,0.11mmol),加入0.3滴乙酸,室温反应15min,加入三乙酰基硼氢化钠(200mg,0.94mmol),室温反应1h,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品2-氯-4-(3-(2-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈55mg,收率:56.1%,LC/MS(ESI+)calcd for C48H52ClFN8O6([M+H]+)m/e 891.3,1H NMR(400MHz,CDCl3)δ8.03(s,1H),7.80(d,J=8.8Hz,1H),7.57(d,J=8.7Hz,1H),7.36(d,J=10.5Hz,1H),6.98(d,J=2.4Hz,1H),6.87–6.76(m,2H),6.67(d,J=8.9Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.54(s,2H),4.45(d,J=10.1Hz,3H),4.25(s,1H),3.98(s,1H),3.89(s,2H),3.63(t,J=34.0Hz,1H),3.18–2.59(m,7H),2.37(s,3H),2.14(dd,J=15.6,7.6Hz,4H),1.86(s,5H),1.61(s,2H),1.45(s,6H),1.23(s,6H).
91:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬-7-基)-N-(甲基-d3)哒嗪-3-甲酰胺
1、7-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬烷-2-羧酸叔丁酯的合成
将2,7-二氮杂螺[3.5]壬-2-羧酸叔丁酯(113mg,0.5mmol)溶于6mL DMF,加入无水碳酸钾(207mg,1.5mmol),加入6-氯-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺(196mg,0.5mmol)、80℃反应过夜,冷却至室温,加入15mL乙酸乙酯,10mL水洗涤三次,饱和食盐水洗涤一次,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品7-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬烷-2-羧酸叔丁酯190mg,收率:65.4%,LC/MS(ESI+)called for C30H37ClN6O4([M+H]+)m/e581.2。
2、叔丁基7-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)(甲基-d3)氨甲酰)哒嗪-3-基)-2,7-二氮杂螺[3.5]壬-2-羧酸酯的合成
将7-{6-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-哒嗪-3-基}-2,7-二氮杂-螺环[3.5]壬烷-2-羧酸叔丁酯(58mg,0.1mmol)溶于5mLDMF,加入氢化纳(8mg,0.2mmol),rt搅拌10min,加入氘代碘甲烷(17mg,0.12mmol),室温反应2h,加水淬灭反应,15mL乙酸乙酯萃取,有机层分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品叔丁基7-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)(甲基-d3)氨甲酰)哒嗪-3-基)-2,7-二氮杂螺[3.5]壬-2-羧酸酯58mg,收率:97.15%,LC/MS(ESI+)calcd forC31H36D3ClN6O4([M+H]+)m/e 598.3。
3、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬-7-基)-N-(甲基-d3)哒嗪-3-甲酰胺的合成
将叔丁基7-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)(甲基-d3)氨甲酰)哒嗪-3-基)-2,7-二氮杂螺[3.5]壬-2-羧酸酯(57mg,0.095mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体30mg,将得到的产品溶于5mL二氯甲烷,加入1-[2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧-2,3-二氢-1H-异吲哚-5-基]-哌啶-4-卡醛(30mg,0.078mmol),加入0.5滴乙酸,室温反应15min,加入三乙酰基硼氢化钠(106mg,0.5mmol),室温反应过夜,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬-7-基)-N-(甲基-d3)哒嗪-3-甲酰胺40mg,收率:48.4%,LC/MS(ESI+)calcd forC45H46D3ClFN9O6([M+H]+)m/e 869.3,1H NMR(400MHz,CDCl3)δ8.53(s,1H),7.64(t,J=10.8Hz,1H),7.55(dd,J=8.7,5.2Hz,1H),7.46(d,J=11.0Hz,1H),7.38(d,J=7.3Hz,1H),7.03–6.92(m,2H),6.83(ddd,J=18.8,8.8,2.1Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.24(s,1H),3.77–3.58(m,6H),3.28(s,3H),2.94–2.68(m,5H),2.57(d,J=5.2Hz,2H),2.25(d,J=8.6Hz,1H),2.15(dd,J=16.3,8.4Hz,2H),2.07(d,J=7.6Hz,1H),1.98(d,J=10.1Hz,1H),1.88(d,J=13.0Hz,6H),1.82–1.64(m,5H),1.58–1.39(m,3H),1.28–1.23(m,1H).
92:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-甲酰胺
1、6-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-羧酸叔丁酯的合成
将2,6-二氮杂螺[3.3]庚烷-2-羧酸叔丁酯(182mg,0.75mmol)溶于5mLDMF,加入6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(196mg,0.5mmol),80℃反应3h,冷却至室温,加入15mL乙酸乙酯,分别用水洗、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得产品6-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-羧酸叔丁酯220mg,收率:79.6%,LC/MS(ESI+)calcd for C28H33ClN6O4([M+H]+)m/e 553.2。
2、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-甲酰胺的合成
将6-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-羧酸叔丁酯(220mg,0.4mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体210mg,将得到的产品(45mg,0.1mmol)溶于5mL二氯甲烷,加入3-[2-(2,6-二氧基哌啶-3-基)-6-氟-1,3-二氧基-2,3-二氢-1H-异吲哚啉-5-基]-3-氮杂-双环[3.1.0]己烷-6-甲醛(37mg,0.096mmol),加入0.5滴乙酸,室温搅拌5min,加入三乙酰基硼氢化钠(212mg,1.0mmol),室温搅拌过夜,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-甲酰胺50mg,收率:60.8%,LC/MS(ESI+)calcd for C42H41ClFN9O6([M+H]+)m/e 822.3,1H NMR(400MHz,CDCl3)δ8.39(s,1H),8.01(d,J=9.2Hz,1H),7.86(d,J=8.1Hz,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=12.5Hz,1H),7.03(d,J=7.4Hz,1H),7.00(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),6.63(d,J=9.2Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.42–4.28(m,4H),4.05(d,J=8.0Hz,1H),3.87(d,J=7.9Hz,2H),3.77(s,3H),3.57(d,J=9.9Hz,2H),2.96–2.70(m,3H),2.70–2.52(m,2H),2.25–2.07(m,5H),1.72–1.67(m,6H),1.46(dd,J=22.5,10.3Hz,3H).
93:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-1,4-二氮杂-1-基)哒嗪-3-羧酰胺
合成步骤参照92
LC/MS(ESI+)calcd for C42H43ClFN9O6([M+H]+)m/e 824.3,1H NMR(400MHz,CDCl3)δ8.27(s,1H),8.01(d,J=9.5Hz,1H),7.88(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.40(s,1H),7.01(dd,J=7.5,4.9Hz,2H),6.91–6.82(m,2H),4.91(dd,J=12.2,5.3Hz,1H),4.37–4.25(m,1H),4.05(dd,J=11.5,7.4Hz,2H),3.86(dd,J=9.9,2.3Hz,2H),3.74(d,J=5.0Hz,2H),3.57(d,J=9.2Hz,2H),2.81(qdd,J=17.5,15.9,10.0Hz,7H),2.59(s,2H),2.22–2.05(m,6H),1.75–1.64(m,6H),1.54–1.36(m,3H).
94:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺
1、5'-[4-(4-氰基-3-三氟甲基苯氧基)-环己基氨甲酰]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯的合成
将N-Boc哌嗪(93mg,0.5mmol)溶于5mL DMF,加入碳酸钾(138mg,1.0mmol),5-氯-吡嗪-2-羧酸[4-(4-氰基-3-三氟甲基-苯氧基)-环己基]-酰胺(142mg,0.33mmol),80℃反应3h,加入15mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到5'-[4-(4-氰基-3-三氟甲基苯氧基)-环己基氨甲酰]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯120mg,收率:63.3%,LC/MS(ESI+)C28H33F3ClN6O4([M+H]+)m/e 574.9。
2、N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺的合成
将5'-[4-(4-氰基-3-三氟甲基苯氧基)-环己基氨甲酰]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯(120mg,0.21mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体100mg。将得到的产品(49mg,0.103mmol)溶于5mL二氯甲烷,加入3-[2-(2,6-二氧基哌啶-3-基)-6-氟-1,3-二氧基-2,3-二氢-1H-异吲哚-5-基]-3-氮杂-双环[3.1.0]己烷-6-甲醛(39mg,0.1mmol),加入0.5滴乙酸,室温搅拌15min,加入三乙酰基硼氢化钠(212mg,1.0mmol),室温反应过夜,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺45mg,收率:50%,LC/MS(ESI+)calcd for C42H41F4ClN9O6([M+H]+)m/e 844.3,1H NMR(400MHz,CDCl3)δ8.86(s,1H),8.24(s,1H),7.97(s,1H),7.74(d,J=8.6Hz,1H),7.45–7.35(m,2H),7.25(d,J=2.1Hz,1H),7.10(dd,J=8.7,2.2Hz,1H),7.04(d,J=7.3Hz,1H),4.92(dd,J=12.0,5.3Hz,1H),4.38(t,J=10.0Hz,1H),4.05(dd,J=18.5,11.1Hz,1H),3.87(dd,J=23.6,6.9Hz,4H),3.60(d,J=9.5Hz,2H),2.95–2.66(m,6H),2.56(d,J=14.1Hz,2H),2.28–2.09(m,5H),1.73–1.64(m,5H),1.56–1.45(m,3H),1.25(dd,J=8.4,5.6Hz,1H),1.10–0.99(m,1H).
95:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺
合成步骤参照94
LC/MS(ESI+)calcd for C42H41F4ClFN9O6([M+H]+)m/e 844.3,1H NMR(400MHz,CDCl3)δ8.27(s,1H),8.05(d,J=9.4Hz,1H),7.88(d,J=8.2Hz,1H),7.75(d,J=8.6Hz,1H),7.40(d,J=12.4Hz,1H),7.26(d,J=2.5Hz,1H),7.11(dd,J=8.6,2.4Hz,1H),7.03(dd,J=11.8,8.6Hz,2H),4.92(dd,J=12.2,5.3Hz,1H),4.39(dd,J=11.9,8.3Hz,1H),4.07(dd,J=11.4,7.5Hz,1H),3.91(d,J=7.7Hz,4H),3.60(d,J=9.0Hz,2H),2.96–2.67(m,6H),2.67–2.44(m,2H),2.28–2.09(m,5H),1.74(dd,J=18.1,8.7Hz,5H),1.55–1.40(m,3H),1.25(dd,J=8.5,5.5Hz,1H),1.06(d,J=11.1Hz,1H).
96:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(6-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-二氮杂螺环[3.4]辛烷-2-基)吡嗪-2-甲酰胺
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2,6-二氮螺环[3.4]辛烷-2-基)吡嗪-2-甲酰胺(50mg,0.11mmol)溶于5mL二氯甲烷,加入1-[2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧-2,3-二氢-1H-异吲哚-5-基]-哌啶-4-卡醛(45mg,0.12mmol),加入0.5滴乙酸,室温反应15min,加入三乙酰基硼氢化钠(),室温反应过夜,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(6-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-二氮杂螺环[3.4]辛烷-2-基)吡嗪-2-甲酰胺35mg,收率:39.0%,LC/MS(ESI+)calcdfor C43H45ClFN9O6([M+H]+)m/e 837.7.
97:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺[3.5]壬-2-基)哒嗪-3-羧酰胺
合成步骤参照88
表征数据LC/MS(ESI+)calcd for C43H45ClFN9O6([M+H]+)m/e 837.7,1H NMR(400MHz,CDCl3)δ8.42–8.26(m,1H),7.99(d,J=9.2Hz,1H),7.88(d,J=8.1Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=10.9Hz,1H),7.39(d,J=7.3Hz,1H),7.00(d,J=2.3Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.60(d,J=9.3Hz,1H),5.00–4.90(m,1H),4.77–4.59(m,2H),4.38–4.26(m,1H),4.12–4.00(m,1H),3.93(s,4H),3.74(d,J=12.0Hz,2H),2.91(d,J=12.2Hz,3H),2.82–2.69(m,4H),2.22–2.13(m,5H),2.04(s,5H),1.82(s,3H),1.69(d,J=12.2Hz,3H),1.47(d,J=12.1Hz,2H).
98:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌啶-4-基)甲基)-2,6-二氮螺环[3.4]辛烷-2-基)哒嗪-3-甲酰胺
合成步骤参照96
表征数据:LC/MS(ESI+)C43H45ClFN9O6([M+H]+)m/e 838。
99:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺
1、5'-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰基]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯
将N-Boc哌嗪(140mg,0.75mmol)溶于5mLDMF,加入无水碳酸钾(207mg,1.5mmol),加入5-氯-吡嗪-2-羧酸[4-(4-氰基-3-氯-苯氧基)-环己基]-酰胺(196mg,0.5mmol),80℃反应3h,加入15mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品5'-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰基]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯130mg,收率:46%,LC/MS(ESI+)called forC27H33ClN6O4([M+H]+)m/e 540.8。
2、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺的合成
将5'-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰基]-2,3,5,6-四氢-[1,2']联吡嗪基-4-羧酸叔丁酯(130mg,0.23mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体100mg,将得到的产品(45mg,0.1mmol)溶于5mL二氯甲烷,加入3-[2-(2,6-二氧基哌啶-3-基)-6-氟-1,3-二氧基-2,3-二氢-1H-异吲哚-5-基]-3-氮杂-双环[3.1.0]己烷-6-甲醛(37mg,0.1mmol),加入0.5滴乙酸,室温反应15min,加入氰基硼氢化钠(80mg,1.3mmol),室温反应过夜,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-甲酰胺20mg,收率:24.7%,LC/MS(ESI+)for C41H41ClFN9O6([M+H]+)m/e 809.8,1H NMR(400MHz,CDCl3)δ8.86(d,J=1.0Hz,1H),8.26(s,1H),7.97(s,1H),7.56(d,J=8.7Hz,1H),7.44–7.36(m,2H),7.04(d,J=7.4Hz,1H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.96–4.86(m,1H),4.34–4.26(m,1H),4.08–3.97(m,1H),3.88(s,2H),3.82(s,3H),3.65–3.53(m,2H),2.73(s,6H),2.58–2.42(m,2H),2.17(d,J=11.9Hz,4H),1.74–1.68(m,6H),1.56–1.38(m,3H).
100:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H44ClN9O6([M+H]+)m/e 817.8,1H NMR(400MHz,CDCl3)δ8.34(s,1H),8.05(d,J=9.5Hz,1H),7.93(d,J=8.2Hz,1H),7.67(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.02(d,J=2.3Hz,1H),6.94(d,J=1.8Hz,1H),6.87(dd,J=8.8,2.4Hz,1H),6.81(d,J=9.0Hz,1H),6.68(d,J=6.9Hz,1H),4.94(dd,J=12.2,5.4Hz,1H),4.35(d,J=4.1Hz,1H),4.09(s,1H),3.78(s,2H),3.68(d,J=10.0Hz,2H),3.48(d,J=8.6Hz,2H),3.36–3.25(m,1H),2.95–2.65(m,6H),2.44(d,J=5.0Hz,1H),2.30(d,J=5.4Hz,1H),2.23–2.10(m,5H),1.87(s,2H),1.71(d,J=12.7Hz,4H),1.55–1.43(m,3H),1.28(d,J=7.8Hz,2H).
101:N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺
1、4-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)哒嗪-3-基)哌嗪-1-羧酸叔丁酯的合成
将N-Boc哌啶酮(60mg,0.38mmol)溶于5mL DMF,加入无水碳酸钾(100mg,0.75mmol),加入6-氯-哒嗪-3-羧酸[3-(3-氯-4-氰基-苯氧基)-2,2,4,4-四甲基-环丁基]-酰胺(105mg,0.25mmol),80℃反应3h,加入15mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品4-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)哒嗪-3-基)哌嗪-1-羧酸叔丁酯85mg,收率:59.7%,LC/MS(ESI+)calcd for C29H37ClN6O4([M+H]+)m/e 568.9。
2、N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺
将4-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)哒嗪-3-基)哌嗪-1-羧酸叔丁酯(85mg,0.15mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体70mg,将得到的产品(35mg,0.07mmol)溶于5mL二氯甲烷,加入3-[2-(2,6-二氧基哌啶-3-基)-6-氟-1,3-二氧基-2,3-二氢-1H-异吲哚-5-基]-3-氮杂-双环[3.1.0]己烷-6-甲醛(30mg,0.077mmol),加入0.5滴乙酸,室温反应15min,加入三乙酰基硼氢化钠(224mmol,1.0mmol),室温反应过夜,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺20mg,收率:34%,LC/MS(ESI+)for C43H45ClFN9O6([M+H]+)m/e 837.7,1H NMR(400MHz,CDCl3)δ8.25–8.12(m,2H),8.06(d,J=9.5Hz,1H),7.59(d,J=8.7Hz,1H),7.42(d,J=12.5Hz,1H),7.06(t,J=8.8Hz,2H),6.99(d,J=2.3Hz,1H),6.83(dd,J=8.7,2.4Hz,1H),4.97–4.88(m,1H),4.21(d,J=9.0Hz,1H),4.09(s,1H),3.93(d,J=7.4Hz,2H),3.62(d,J=9.1Hz,2H),3.08–2.70(m,6H),2.70–2.52(m,2H),2.20–2.11(m,1H),1.76(s,6H),1.29(d,J=12.5Hz,6H),1.23(s,6H),1.07(s,1H),0.88(d,J=20.0Hz,1H).
102:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C41H42ClN9O6([M+H]+)m/e 791.7,1H NMR(400MHz,CDCl3)δ8.26(s,1H),8.07(d,J=9.1Hz,1H),7.90(d,J=8.1Hz,1H),7.67(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.06–7.00(m,2H),6.96(d,J=2.0Hz,1H),6.87(dd,J=8.7,2.4Hz,1H),6.70(dd,J=8.4,2.1Hz,1H),4.96(dd,J=12.2,5.3Hz,1H),4.34(t,J=10.0Hz,1H),4.08(d,J=8.0Hz,1H),3.93(s,2H),3.71(d,J=9.9Hz,2H),3.52(d,J=9.7Hz,2H),2.83(ddd,J=33.3,23.9,10.4Hz,5H),2.20(s,4H),1.73–1.59(m,11H),1.55–1.42(m,3H).
103:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺环[3.4]辛基-2-基)哌啶-1-基)哒嗪-3-甲酰胺
1、6-(4-羟基哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺的合成
将哌啶-4-醇(100mg,1.0mmol)溶于5mL DMF,加入无水碳酸钾(415mg,3.0mmol),加入5-氯-哒嗪-2-羧酸[4-(4-氰基-3-氯-苯氧基)-环己基]-酰胺(390mg,1.0mmol),80℃反应3h,加入15mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品6-(4-羟基哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺310mg,收率:68.3%,LC/MS(ESI+)calcd for C23H26ClN5O3([M+H]+)m/e 455.9。
2、6-(4-氧代哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺将6-(4-羟基哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺(250mg,0.55mmol)溶解于10mL二氯甲烷,加入Dess-Martin氧化剂(280mg,0.66mmol),室温搅拌反应3h,过滤,滤液减压浓缩干,薄层色谱层析分离,得到产物6-(4-氧代哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺180mg,收率:72%,LC/MS(ESI+)calcd forC23H24ClN5O3([M+H]+)m/e453.9。
3、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺环[3.4]辛基-2-基)哌啶-1-基)哒嗪-3-甲酰胺
将6-(4-氧代哌啶-1-基)-哒嗪-3-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺(46mg,0.1mmol)、5-(2,6-二氮杂螺[3.4]辛-6-基)-2-(2,6-二氧哌啶-3-基)-6-氟异吲哚啉-1,3-二酮()溶解于5mL二氯甲烷中,加入1滴乙酸,室温反应15min,加入氰基硼氢化钠(80mg,1.3mmol),室温反应过夜,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺环[3.4]辛基-2-基)哌啶-1-基)哒嗪-3-甲酰胺16mg,收率:19.5%,LC/MS(ESI+)for C42H43ClFN9O6([M+H]+)m/e 823.8,1H NMR(400MHz,CDCl3)δ8.14–8.06(m,1H),8.06–8.01(m,1H),7.91–7.85(m,1H),7.61–7.56(m,1H),7.47–7.41(m,1H),7.03(s,3H),6.90–6.85(m,1H),4.94(dd,J=12.2,5.3Hz,1H),4.53(s,1H),4.34(s,1H),4.07(s,1H),3.81(d,J=4.5Hz,2H),3.65(s,2H),3.20(s,2H),2.93–2.75(m,3H),2.19(d,J=11.1Hz,5H),1.99(d,J=6.6Hz,2H),1.71(d,J=12.4Hz,2H),1.61(s,9H),1.52(dd,J=18.4,7.6Hz,3H).
104:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)m/e 791.8for C42H43ClN9O6([M+H]+).1H NMR(400MHz,CDCl3)δ8.88(s,1H),8.34–8.24(m,1H),8.04–7.96(m,1H),7.72–7.64(m,1H),7.62–7.55(m,1H),7.47–7.38(m,1H),7.02(s,1H),6.99–6.93(m,1H),6.90–6.83(m,1H),6.74–6.66(m,1H),4.95(dd,J=12.1,5.2Hz,1H),4.32(dd,J=12.9,6.8Hz,1H),4.06(d,J=10.0Hz,2H),3.88(s,3H),3.70(d,J=9.9Hz,2H),3.53(d,J=8.5Hz,2H),2.98–2.69(m,6H),2.58(s,2H),2.29–2.09(m,5H),1.73(dd,J=19.2,9.9Hz,5H),1.55–1.45(m,3H).
105:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)-1,3,4-噻二唑-2-甲酰胺
1、乙基5-(7-(叔丁氧羰基)-2,7-二氮杂螺[3.5]壬-2-基)-1,3,4-噻二唑-2-羧酸酯的合成
将2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯(249mg,1.1mmol)溶于6mL DMF,加入无水碳酸钾(416mg,3.0mmol),加入5-氯-1,3,4-噻二唑-2-羧酸乙酯(193mg,1.0mmol),80℃反应1.5h,冷却至室温,加入20mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品乙基5-(7-(叔丁氧羰基)-2,7-二氮杂螺[3.5]壬-2-基)-1,3,4-噻二唑-2-羧酸酯295mg,收率:79.9%,LC/MS(ESI+)called forC17H26N2O2S([M+H]+)m/e 369.1。
2、5-(2,7-二氮杂螺[3.5]壬-2-基)-[1,3,4]噻二唑-2-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺的合成
将4-(4-氨基环己氧基)-2-氯-苯甲腈(210mg,0.84mmol)溶解于5mL甲醇,加入2-(5-乙氧羰基-[1,3,4]噻二唑-2-基)-2,7-二氮杂螺环[3.5]壬-7-羧酸第三丁酯(268mg,0.7mmol),85℃回流过夜,薄层色谱层析分离,得到白色固体产品360mg,将其溶解于5mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2.5h,减压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体产品5-(2,7-二氮杂螺[3.5]壬-2-基)-[1,3,4]噻二唑-2-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺290mg,收率:85%,LC/MS(ESI+)called for C23H27ClN6O2S([M+H]+)m/e 487.1。
3、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)-1,3,4-噻二唑-2-甲酰胺的合成
将5-(2,7-二氮杂螺[3.5]壬-2-基)-[1,3,4]噻二唑-2-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺(49mg,0.1mmol)、1-[2-(2,6-二氧基哌啶-3-基)-1,3-二氧基-2,3-二氢-1H-异吲哚-5-基]-哌啶-4-乙醛(37mg,0.1mmol)加入圆底烧瓶中,加入5mL二氯甲烷,加入1滴乙酸催化反应,氩气保护下,室温搅拌10min,加入三乙酰基硼氢化钠(212mg,1.0mmol),室温反应3h,水洗两次,有机层再用饱和食盐水洗涤一次,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)-1,3,4-噻二唑-2-甲酰胺45mg,收率:53.6%,LC/MS(ESI+)C42H46ClN9O6S([M+H]+)m/e 840.3.1H NMR(400MHz,CDCl3)δ8.14(s,1H),7.67(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.27(s,1H),7.08–6.95(m,3H),6.84(dd,J=8.8,2.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.30(t,J=10.0Hz,1H),3.96(dd,J=26.6,12.1Hz,6H),3.07–2.66(m,6H),2.35(s,2H),2.18(s,6H),1.88(s,4H),1.65(d,J=17.7Hz,7H),1.53–1.44(m,2H),1.25(dd,J=8.5,5.5Hz,3H).
106:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)嘧啶-2-甲酰胺
1、2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬烷-7-羧酸叔丁酯的合成
将5-溴-嘧啶-2-羧酸[4-(3-氯-4-氰基-苯氧基)-环己基]-酰胺(109mg,0.25mmol)溶解于5mL 1,4-二氧六环,加入2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯(57mg,0.25mmol),BINAP(8mg,12.5μmol),Cs2CO3(163mg,0.5mmol),Pd(OAc)2(3mg,12.5μmol),氩气保护,110℃回流过夜,加入乙酸乙酯15mL,用10mL水洗涤三次,饱和食盐水洗涤一次,无水硫酸钠干燥,减压浓缩干,波层色谱层析分离,得到产物2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬烷-7-羧酸叔丁酯100mg,收率:68.9%,LC/MS(ESI+)called for C30H37ClN6O4([M+H]+)m/e 580.9。
2、4-(2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯的合成
将2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬烷-7-羧酸叔丁酯(100mg,0.17mmol)溶解于5mL二氯甲烷,加入3mL三氟乙酸,室温搅拌3h,加压浓缩干,用饱和Na2CO3水溶液调节pH至碱性,DCM萃取2次,合并有机层,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到白色固体80mg。将得到的产品(78mg,0.16mmol)溶于5mL二氯甲烷,加入4-Boc哌啶酮(199mg,0.98mmol),加入0.5滴乙酸,40℃回流1h,冷却至室温,加入三乙酰基硼氢化钠(424mg,2.0mmol),40℃回流反应5h,冷却至室温,水洗,减压浓缩干,薄层色谱层析分离,得到产品4-(2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯50mg,收率:47.0%,LC/MS(ESI+)called for C35H46ClN7O4([M+H]+)m/e 664.3。
3、N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异戊醇-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)嘧啶-2-甲酰胺的合成
将4-(2-{2-[4-(3-氯-4-氰基-苯氧基)-环己基氨甲酰]-嘧啶-5-基}-2,7-二氮杂-螺环[3.5]壬-7-基)-哌啶-1-羧酸叔丁酯(50mg,0.075mmol)溶解于3mL二氯甲烷,加入3mL三氟乙酸,室温搅拌2h,减压浓缩干,加入二氯甲烷再减压浓缩2次,加入3mL DMSO,加入DIEA 0.5mL,加入2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(30mg,0.11mmol),135℃回流1.5h,冷却至室温,加入10mL二氯甲烷,10mL水洗涤2次,再用饱和食盐水洗涤1次,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离,得到黄色固体产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-2-基)嘧啶-2-甲酰胺26mg,收率:42.3%,LC/MS(ESI+)m/e820.3for C43H46ClN9O6([M+H]+).1H NMR(400MHz,CDCl3)δ8.16(s,1H),7.95(s,2H),7.69(d,J=8.5Hz,2H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.1Hz,1H),7.06(dd,J=8.6,2.1Hz,1H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.99–4.90(m,1H),4.34–4.25(m,1H),4.15–4.07(m,1H),4.02(d,J=13.3Hz,2H),3.80(s,4H),3.00(t,J=12.0Hz,2H),2.95–2.71(m,4H),2.25–2.13(m,7H),2.01(s,4H),1.69(d,J=12.3Hz,8H),1.46(dd,J=24.4,11.7Hz,2H).
107:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬烷-2-基)-1,3,4-噻二唑-2-甲酰胺
合成步骤参照105
表征数据:LC/MS(ESI+)m/e 858.3for C42H45ClFN9O6S([M+H]+),1H NMR(400MHz,CDCl3)δ8.14–8.08(m,1H),7.56(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.39(d,J=7.3Hz,1H),7.03(d,J=8.2Hz,1H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),4.97–4.90(m,1H),4.36–4.26(m,1H),4.05–3.95(m,1H),3.90(s,4H),3.73(q,J=7.0Hz,3H),3.65(d,J=12.1Hz,2H),2.95–2.70(m,5H),2.40–2.34(m,2H),2.22(d,J=7.6Hz,2H),1.92–1.86(m,6H),1.69–1.61(m,4H),1.50(dd,J=18.4,7.7Hz,2H),1.25(t,J=7.0Hz,5H).
108:N-(1-(3-氯-4-氰基苯)哌啶-4-基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1)2-氯-4-甲酰苯甲腈
将4-溴-2-氯-苯甲腈(5.0g,23.1mmol)溶解于10mL无水四氢呋喃,氩气保护,冰水浴,滴加15mL 2N异丙基溴化镁/THF溶液,滴加完毕后,保温搅拌1h,加入哌啶-1-乙醛(7.75g,68.5mmol),保温反应2h,加入15mL饱和氯化铵水溶液淬灭反应,20mL乙酸乙酯萃取,水层再用20mL乙酸乙酯反萃一次,合并有机层,有机层用饱和食盐水洗涤一次,无水硫酸钠干燥,过滤,滤液减压浓缩干,色谱柱层析分离纯化,得到产品2-氯-4-甲酰苯甲腈2.22g,收率:58%。2)(1-(3-氯-4-氰基苯)哌啶-4-基)氨基甲酸叔丁酯
将2-氯-4-甲酰苯甲腈(820mg,5.0mmol)、哌啶-4-基氨基甲酸叔丁基酯(1.0g,5.0mmol)溶解于15mL无水甲醇,加入2滴乙酸,60℃反应1h,冷却至室温,加入氰基硼氢化钠(942mg,15mmol),反应过夜,加入20mL丙酮,搅拌20min,减压浓缩干,加入20mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩干,色谱柱层析分离纯化,得到产品(1-(3-氯-4-氰基苯)哌啶-4-基)氨基甲酸叔丁酯680mg,收率:38.9%,LC/MS(ESI+)called for C18H24ClN3O2([M+H]+)m/e 350.2。
3)6-氯-N-(1-(3-氯-4-氰基苯)哌啶-4-基)哒嗪-3-甲酰胺
将(1-(3-氯-4-氰基苯)哌啶-4-基)氨基甲酸叔丁酯(370mg,0.95mmol)溶于10mL二氯甲烷,加入8mL三氟乙酸,室温搅拌2h,减压浓缩干,饱和Na2CO3水溶液调节pH至碱性,乙酸乙酯萃取,有机层用无水硫酸钠干燥,减压浓缩干,得到产品230mg。将6-氯吡啶-3-羧酸(145mg,0.91mmol)溶于5mL二氯甲烷,加入N,N-二异丙基乙胺(235mg,1.82mmol),冰水浴下加入HATU(533mg,1.38mmol),将上一步的化合物溶解于5mL二氯甲烷,滴加入反应体系中,滴加完毕后,缓慢升温至室温,反应2h,分别用饱和氯化铵水溶液、饱和食盐水洗涤,无水硫酸镁干燥,过滤,减压浓缩干,薄层色谱层析分离纯化,得到产品6-氯-N-(1-(3-氯-4-氰基苯)哌啶-4-基)哒嗪-3-甲酰胺200mg,收率:53.7%,LC/MS(ESI+)called for C18H17Cl2N5O([M+H]+)m/e 390。
4)4-((1-(6-((1-(3-氯-4-氰基苯)哌啶-4-基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯
将4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(71mg,0.25mmol)溶于5mL DMF,加入无水碳酸钾(102mg,0.75mmol),加入6-氯-N-(1-(3-氯-4-氰基苯)哌啶-4-基)哒嗪-3-甲酰胺(100mg,0.25mmol),80℃反应3h,冷却至室温,加入20mL乙酸乙酯,分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离,得到产品4-((1-(6-((1-(3-氯-4-氰基苯)哌啶-4-基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯140mg,收率:87.9%,LC/MS(ESI+)called for C33H45ClN8O3([M+H]+)m/e 637.3。
5)N-(1-(3-氯-4-氰基苯)哌啶-4-基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
将4-((1-(6-((1-(3-氯-4-氰基苯)哌啶-4-基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(75mg,0.12mmol)溶于3mL二氯甲烷,加入2mL三氟乙酸,室温搅拌2h,减压浓缩干,加入5mL DMSO,加入0.8mL DIEA,加入2-(2,6-二氧哌啶-3-基)-5-氟异吲哚啉-1,3-二酮(60mg,0.22mmol),140℃反应3h,冷却至室温,加入15mL二氯甲烷,饱和氯化铵洗涤2次,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,薄层色谱层析分离纯化,得到产品N-(1-(3-氯-4-氰基苯)哌啶-4-基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺42mg,收率:44.1%,LC/MS(ESI+)for C41H45ClN10O5([M+H]+)m/e 793.3,1H NMR(400MHz,CDCl3)δ8.12(s,1H),7.97(d,J=9.5Hz,1H),7.90(d,J=8.0Hz,1H),7.70(d,J=8.5Hz,1H),7.63(d,J=7.9Hz,1H),7.54(s,1H),7.37(d,J=8.5Hz,1H),7.29(d,J=2.0Hz,1H),7.06(dd,J=8.6,2.0Hz,1H),6.98(d,J=9.6Hz,1H),4.95(dd,J=12.2,5.2Hz,1H),4.52(d,J=13.0Hz,2H),4.08–3.95(m,1H),3.55(s,2H),3.44(s,4H),3.05(t,J=12.0Hz,2H),2.95–2.72(m,5H),2.60(s,3H),2.28(d,J=11.0Hz,4H),2.14(dd,J=7.6,5.3Hz,1H),1.99(dd,J=24.5,13.5Hz,5H),1.36–1.16(m,5H).
109:N-(6-(3-氯-4-氰基苯氧基)螺环[3.3]庚烷-2-基)-6-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C42H44ClN9O6([M+H]+)m/e 806.2,1H NMR(400MHz,CDCl3)δ8.09(s,1H),8.05(d,J=8.1Hz,1H),7.95(d,J=9.6Hz,1H),7.71(d,J=8.5Hz,1H),7.55(d,J=8.7Hz,1H),7.30(d,J=2.1Hz,1H),7.07(dd,J=8.6,2.1Hz,1H),6.97(d,J=9.7Hz,1H),6.88(d,J=2.3Hz,1H),6.75(dd,J=8.7,2.4Hz,1H),4.95(dd,J=12.3,5.3Hz,1H),4.66–4.59(m,1H),4.56–4.51(m,2H),3.49(s,3H),3.06(t,J=12.1Hz,2H),2.98–2.74(m,4H),2.75–2.67(m,2H),2.65–2.57(m,2H),2.56–2.49(m,2H),2.26(ddd,J=24.1,11.9,6.9Hz,4H),2.18–2.08(m,3H),1.99(d,J=12.8Hz,3H),1.50–1.40(m,1H),1.29(dd,J=20.4,11.2Hz,4H)。
110:N-((1r,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3S)-1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)吡咯烷-3-基)氧基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd forC40H40ClFN8O7([M+H]+)m/e 799.2,1H NMR(400MHz,CDCl3)δ8.06(d,J=6.8Hz,2H),7.92(d,J=7.8Hz,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=12.3Hz,1H),7.13(d,J=9.7Hz,1H),7.04(d,J=7.5Hz,1H),7.00(d,J=2.3Hz,1H),6.86(dd,J=8.7,2.3Hz,1H),4.92(dd,J=12.2,5.3Hz,1H),4.33(dd,J=21.3,12.3Hz,2H),4.05(d,J=3.7Hz,2H),3.84–3.58(m,6H),2.96–2.69(m,3H),2.15(t,J=13.5Hz,6H),1.98(s,2H),1.78(s,2H),1.67(dd,J=21.9,10.8Hz,4H),1.48(dd,J=22.3,11.5Hz,3H).
111:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((3R)-1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)吡咯烷-3-基)氧基)哌啶-1-基)苯甲酰胺
1)(S)-3-((1-(4-(甲氧羰基)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸叔丁酯将(S)-3-(哌啶-4-氧基)吡咯烷-1-羧酸叔丁酯(260mg,0.96mmol)溶解于5mL DMSO,加入碳酸钾(420mg,3.0mmol),加入对氟苯甲酸甲酯(308mg,2.0mmol),100℃反应3h,冷却至室温,加入15mL乙酸乙酯,分别用水、食盐水洗涤,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品(S)-3-((1-(4-(甲氧羰基)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸叔丁酯130mg,收率:33.5%,LC/MS(ESI+)called for C22H32N2O5([M+H]+)m/e 405.3。
2)(R)-3-((1-(4-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸叔丁酯
将(S)-3-((1-(4-(甲氧羰基)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸叔丁酯(130mg,0.32mmol)溶于5mL四氢呋喃,加入5mL 2N NaOH水溶液,室温搅拌过夜,用0.5N HCl调节pH=5,加入15mL乙酸乙酯,分别用水、食盐水洗涤,无水硫酸钠干燥,减压浓缩干,得到产品。将其溶于5mL二氯甲烷,加入DIEA(101mg,1.0mmol),加入HATU(141mg,0.37mmol),室温搅拌10min,加入4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(80mg,0.32mmol),室温反应过夜,分别用饱和氯化铵水溶液、水、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品(R)-3-((1-(4-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸叔丁酯160mg,收率:80.26%,LC/MS(ESI+)called for C34H43ClN4O5([M+H]+)m/e 623.2.
3)N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((3R)-1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)吡咯烷-3-基)氧基)哌啶-1-基)苯甲酰胺
将叔丁基(R)-3-((1-(4-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)苯基)哌啶-4-基)氧基)吡咯烷-1-羧酸盐(50mg,0.080mmol)溶于3mL二氯甲烷,加入2mL三氟乙酸,室温搅拌2h,减压浓缩干,加入5mL DMSO,加入0.8mL DIEA,加入2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(23mg,0.080mmol),140℃反应3h,冷却至室温,加入15mL二氯甲烷,饱和氯化铵洗涤2次,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((3R)-1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)吡咯烷-3-基)氧基)哌啶-1-基)苯甲酰胺36mg,收率:57.8%,LC/MS(ESI+)for C42H43ClN6O7([M+H]+)m/e 779.2,1H NMR(400MHz,CDCl3)δ8.13(s,1H),7.67(d,J=8.4Hz,3H),7.56(d,J=8.7Hz,1H),6.99(d,J=2.4Hz,1H),6.95(t,J=6.0Hz,2H),6.84(dd,J=8.8,2.4Hz,1H),6.69(dd,J=8.5,2.1Hz,1H),5.90(d,J=7.5Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.62(s,1H),4.42(s,1H),4.33–4.23(m,1H),4.08–4.00(m,1H),3.60(ddd,J=22.8,13.9,7.0Hz,5H),3.53–3.46(m,1H),3.43(d,J=9.2Hz,1H),3.17–3.03(m,2H),2.81(tdd,J=19.5,17.2,10.4Hz,3H),2.26–2.10(m,7H),2.01(s,2H),1.64(dd,J=13.1,2.9Hz,3H),1.51–1.30(m,3H).
112:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-咪唑-4-羧酰胺
1)N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-咪唑-4-甲酰胺
将1H-咪唑-4-羧酸(448mg,4.0mmol)溶解于10mL DMF,加入DIEA(1.03g,8.0mmol),HATU(1.52g,4.0mmol),室温搅拌30min,加入4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(1.0g,4.0mmol),室温搅拌过夜,加入25mL乙酸乙酯,20mL水洗涤,有机层再分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,色谱柱层析分离纯化,得到产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-咪唑-4-甲酰胺610mg,收率:44.2%,LC/MS(ESI+)called for C17H17ClN4O2([M+H]+)m/e 345.1。
2)(1R,4r)-4-(4-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-咪唑-1-基)环己烷-1-羧酸甲酯
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-咪唑-4-甲酰胺(345mg,1.0mmol)溶解于6mL DMF,加入碳酸铯(975mg,3.0mmol),室温搅拌5min,加入甲基(1r,4r)-4-((甲磺酰基)氧基)环己烷-1-羧酸酯(354mg,4.5mmol),95℃反应过夜,加入25mL乙酸乙酯,20mL水洗涤,有机层再分别用水、饱和食盐水洗涤,无水硫酸钠干燥,过滤,减压浓缩干,薄层色谱层析分离纯化,得到产品甲基(1R,4r)-4-(4-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-咪唑-1-基)环己烷-1-羧酸酯130mg,收率:26.8%,LC/MS(ESI+)called for C25H29ClN4O4([M+H]+)m/e 485.3。
3)N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-咪唑-4-甲酰胺
将甲基(1R,4r)-4-(4-((1R,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-咪唑-1-基)环己烷-1-羧酸酯(130mg,0.27mmol)溶解于5mL四氢呋喃,加入4mL甲醇,加入硼氢化钠(76mg,2.0mmol),70℃反应过夜,减压浓缩干,加入10mL乙酸乙酯,水洗,饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-咪唑-4-甲酰胺54mg,收率:43.8%,LC/MS(ESI+)calcd for C24H29ClN4O43([M+H]+)m/e 457.3。
4)N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-咪唑-4-甲酰胺
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-咪唑-4-甲酰胺(54mg,0.12mmol)溶解于5mL二氯甲烷,加入Dess-Martin氧化剂(102mg,0.24mmol),室温搅拌1h,过滤,滤液减压浓缩干,得到产品N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-咪唑-4-甲酰胺45mg,收率:84.5%,LC/MS(ESI+)called for C24H27ClN4O3([M+H]+)m/e 455.2。
5)N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-咪唑-4-羧酰胺
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-咪唑-4-甲酰胺(30mg,0.066mmol)、2-(2,6-二氧哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚啉-1,3-二酮(26mg,0.066mmol)溶解于5mL二氯甲烷,加入0.5滴乙酸,室温搅拌10min,加入三乙酰基硼氢化钠(212mg,1.0mmol),室温搅拌3h,水洗,饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-咪唑-4-羧酰胺16mg,收率:28.9%,LC/MS(ESI+)for C44H48ClFN8O6([M+H]+)m/e 839.2,1H NMR(400MHz,CDCl3)δ7.66(s,1H),7.57(d,J=8.8Hz,1H),7.51(s,1H),7.37(d,J=10.9Hz,1H),7.01(d,J=2.3Hz,1H),6.86(dd,J=8.8,2.3Hz,1H),6.82(d,J=7.5Hz,1H),4.91(dd,J=12.1,5.3Hz,1H),4.30(d,J=10.3Hz,1H),3.98(d,J=11.1Hz,2H),3.92(s,4H),2.92–2.65(m,4H),2.14(dd,J=17.2,7.5Hz,8H),2.05(s,2H),1.95(s,4H),1.67(dd,J=25.5,12.2Hz,6H),1.53–1.40(m,3H),1.29(s,2H),1.17(s,2H).
113:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-咪唑-4-甲酰胺
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-咪唑-4-甲酰胺(15mg,0.033mmol)、2-(2,6-二氧哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(13mg,0.033mmol)溶解于5mL二氯甲烷,加入0.3滴乙酸,室温搅拌10min,加入三乙酰基硼氢化钠(106mg,0.5mmol),室温搅拌3h,水洗,饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-咪唑-4-甲酰胺13mg,收率:47.0%,LC/MS(ESI+)for C44H49ClN8O6([M+H]+)m/e 821.2,1H NMR(400MHz,CDCl3)δ8.58(s,1H),7.64(dd,J=13.2,4.7Hz,2H),7.55(d,J=8.7Hz,1H),7.50(d,J=1.3Hz,1H),7.06(d,J=8.3Hz,1H),6.98(d,J=2.4Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),6.78(d,J=1.9Hz,1H),6.51(d,J=8.0Hz,1H),4.94(dd,J=12.3,5.2Hz,1H),4.27(t,J=10.2Hz,1H),4.08–3.89(m,2H),3.76(s,4H),2.96–2.64(m,4H),2.38(s,2H),2.15(d,J=10.2Hz,8H),1.95(s,2H),1.77–1.59(m,13H),1.45(dd,J=23.1,11.0Hz,3H).
114:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-1,2,3-三唑-4-羧酰胺
合成步骤参照112
表征数据:LC/MS(ESI+)for C43H48ClN9O6([M+H]+)m/e 822.3,1H NMR(400MHz,CDCl3)δ8.09(d,J=16.2Hz,2H),7.68(d,J=6.0Hz,1H),7.56(d,J=8.7Hz,1H),7.08(d,J=8.0Hz,1H),7.00(d,J=2.1Hz,1H),6.85(dd,J=8.8,2.3Hz,1H),6.80(s,1H),6.56(s,1H),4.94(d,J=6.3Hz,1H),4.48(s,1H),4.31(s,1H),4.05(d,J=7.9Hz,1H),3.85(s,4H),2.96–2.69(m,7H),2.33(s,4H),2.24–2.02(m,8H),1.89(d,J=8.2Hz,2H),1.67(d,J=10.4Hz,8H),1.53–1.46(m,2H).
115:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-1H-1,2,3-三唑-4-甲酰胺
LC/MS(ESI+)for C43H47ClFN9O6([M+H]+)m/e 840.3,1H NMR(400MHz,CDCl3)δ8.10(s,1H),8.08(d,J=4.4Hz,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=10.7Hz,1H),7.07(d,J=8.2Hz,1H),7.00(d,J=2.3Hz,1H),6.89–6.80(m,2H),4.96–4.88(m,1H),4.53–4.42(m,1H),4.31(s,1H),4.05(d,J=8.3Hz,1H),4.01(d,J=25.7Hz,4H),3.63(s,2H),3.01–2.60(m,7H),2.34(d,J=12.8Hz,3H),2.17(dd,J=17.1,12.1Hz,7H),1.97–1.82(m,2H),1.67(d,J=12.9Hz,2H),1.62(s,6H),1.48(d,J=12.2Hz,2H).
116:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-2-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-2H-1,2,3-三唑-4-羧酰胺
LC/MS(ESI+)for C43H48ClN9O6([M+H]+)m/e 822.3,1H NMR(400MHz,CDCl3)δ8.09–8.04(m,1H),8.03(s,1H),7.66(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),6.99(d,J=2.4Hz,1H),6.88–6.82(m,1H),6.79(d,J=2.0Hz,1H),6.61–6.56(m,1H),6.56–6.50(m,1H),4.94(dd,J=12.2,5.3Hz,1H),4.46(dd,J=13.8,10.0Hz,1H),4.30(s,1H),4.04(d,J=7.9Hz,1H),3.79(s,4H),2.95–2.71(m,4H),2.58(s,2H),2.28(d,J=11.1Hz,2H),2.24–2.10(m,7H),1.98(d,J=13.5Hz,3H),1.69(d,J=10.0Hz,8H),1.55–1.43(m,2H),1.25(s,3H).
117:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-2-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)环己基)-2H-1,2,3-三唑-4-甲酰胺
LC/MS(ESI+)for C43H47ClFN9O6([M+H]+)m/e 840.3,1H NMR(400MHz,CDCl3)δ8.03(s,1H),7.98(s,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=10.8Hz,1H),7.00(d,J=2.4Hz,1H),6.87–6.80(m,2H),6.58(d,J=8.2Hz,1H),4.92(dd,J=12.3,5.2Hz,1H),4.46(t,J=11.9Hz,1H),4.31(t,J=9.9Hz,1H),4.04(d,J=7.8Hz,1H),3.96(s,3H),2.83(ddd,J=28.3,24.3,14.8Hz,4H),2.29(d,J=10.8Hz,2H),2.15(dd,J=20.4,8.2Hz,6H),2.04–1.96(m,2H),1.67(d,J=22.9Hz,10H),1.52–1.45(m,2H),1.25(s,4H).
118:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(叔丁氧酰胺-哌啶-1-基)哒嗪-3-酰胺
化合物N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-氯哒嗪-3-羧酰胺(500mg,1.28mmol),4-叔丁氧羰基氨基哌啶(384mg,1.92mmol)和N,N-二异丙基乙胺(496mg,3.84mmol)加入到10mL DOX中,加热到110℃,搅拌过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层分别用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(叔丁氧酰胺)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺(493mg,0.87mmol),收率:68%。
LC/MS(ESI+)calcd for C28H35ClN6O4 +(M+H+)m/z,555.2;found555.2。
2.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氨基哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(叔丁氧酰胺)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-羧酰胺(200mg,0.36mmol)溶于3mL二氯甲烷,加入6mL三氟醋酸,在室温下搅拌2h,减压蒸馏浓缩后加入饱和碳酸钠溶液调节pH 10后用二氯甲烷萃取。有机层用食盐水洗涤,无水硫酸钠干燥,旋干。得到淡黄色油状物化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氨基哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺(159mg,0.35mmol),收率:97%。LC/MS(ESI+)calcd for C23H27ClN6O2 +(M+H+)m/z,455.2;found455.2.
3.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(4-叔丁基氧基氧代氨基-哌啶-1-基)-(4-氨基哌啶-1-基)哒嗪-3-酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氨基哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺(150mg,0.33mmol)和N-叔丁氧羰基-4-哌啶酮(131mg.0.66mmol)溶于1mL甲醇和2mL二氯甲烷的混合溶剂之中,在两滴冰醋酸的催化之下,室温搅拌半个小时后,加入三乙酰氧基硼氢化钠(91mg,0.43mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(4-叔丁基氧基氧代氨基-哌啶-1-基)-(4-氨基哌啶-1-基)哒嗪-3-酰胺(60mg,0.094mmol),收率:28%。LC/MS(ESI+)calcd for C33H44ClN7O4 +(M+H+)m/z,638.3;found638.3.
4.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(4-氨基-哌啶-1-基)-(4-氨基哌啶-1-基)哒嗪-3-酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(4-叔丁基氧基氧代氨基-哌啶-1-基)-(4-氨基哌啶-1-基)哒嗪-3-酰胺(60mg,0.094mmol)溶于3mL二氯甲烷,加入6mL三氟醋酸,在室温下搅拌2h,减压蒸馏浓缩后得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氨基哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺三氟醋酸盐(55mg,0.084mmol),收率:90%。LC/MS(ESI+)calcd for C28H36ClN7O2 +(M+H+)m/z,538.3;found538.3.
5.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-氨基哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺三氟醋酸盐(55mg,0.084mmol),2-(2,6-二氧代哌啶-3-基)-5-氟-1,3-二氧代异吲哚啉(23mg,0.084mmol)和DIPEA(54mg,0.42mmol)加入到1mLDMSO。加热至130℃,搅拌3h。冷却至室温,加水和乙酸乙酯萃取,有机层用饱溶液和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-酰胺(15mg,0.019mmol),收率:23%。LC/MS(ESI+)calcd for C41H44ClN9O6 +(M+H+)m/z,794.3;found794.2.1H NMR(400MHz,DMSO-d6)δ11.06(s,2H),8.48(d,J=2.0Hz,1H),7.84(d,J=4.4Hz,1H),7.69(d,J=4.4Hz,1H),7.64(d,J=4.4Hz,1H),7.37(d,J=1.2Hz,1H),7.34-7.21(m,3H),7.11(dd,J=5.6,3.2Hz,1H),6.84(d,J=4.8Hz,1H),5.05(dd,J=8.8,3.6Hz,1H),4.55-4.47(m,1H),4.12-4.03(m,2H),3.86-3.76(m,2H),2.99-2.91(m,2H),2.89-2.82(m,2H),2.60-2.52(m,1H),2.27-2.20(m,2H),2.11-2.04(m,2H),2.00-1.93(m,3H),1.86-1.82(m,2H),1.82-1.79(m,2H),1.64-1.56(m,2H),1.53-1.38(m,6H),1.23-1.19(m,2H).
119:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)甲氨基)哌啶-1-基)哒嗪-3-酰胺的合成
LC/MS(ESI+)calcd for C42H45ClFN9O6 +(M+H+)m/z,826.3;found826.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.62(d,J=4.0Hz,1H),7.86(d,J=4.4Hz,1H),7.81(d,J=4.8Hz,1H),7.72(d,J=5.6Hz,1H),7.45(d,J=4.0Hz,1H),7.41-7.39(m,1H),7.39-7.34(m,1H),7.17-7.11(m,1H),5.11(dd,J=9.2,3.6Hz,1H),4.59-4.47(m,3H),3.69-3.62(m,2H),3.47-3.41(m,1H),3.32-3.28(m,3H),3.09-2.99(m,2H),2.98-2.87(m,3H),2.55(s,3H),2.28-2.16(m,3H),2.16-2.07(m,2H),2.07-1.96(m,2H),1.96-1.77(m,2H),1.72-1.61(m,4H),1.56-1.46(m,4H),1.08-1.03(m,1H).
120:2-氯-4-((2-(6-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚烷-6-基)氧)苄腈的合成
1.化合物6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯的合成
化合物氢化钠(60%)(514mg,12.86mmol)溶于20mL DMF,在-10~0℃下搅拌10min后,加入6-羟基-2-氮杂螺[3.3]庚烷-2-甲酸叔丁酯(2.05g,9.64mmol),在-10~0℃下搅拌30min后,缓慢滴加2-氯-4-氟苯腈(1.00g,6.43mmol)的DMF溶液,在-10~0℃下搅拌1h后,自然升温至室温后继续搅拌2h。反应液加入水和乙酸乙酯萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到黄色油状化合物叔丁基6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羧酸盐(1.12g,3.21mmol),收率:50%。
LC/MS(ESI+)calcd for C18H21ClN2O3 +(M+H+)m/z,349.1;found349.1.
2.化合物4-(2-氮杂螺[3.3]庚-6-基氧基)-2-氯苄腈的合成
化合物6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯(348mg,1.00mmol)和2mL三氟醋酸溶于1mL二氯甲烷中,室温搅拌1h后,TLC检测反应完全,浓缩反应液,得到白色固体化合物4-(2-氮杂螺[3.3]庚-6-基氧基)-2-氯苄腈三氟醋酸盐,直接作为下一步反应的原料,收率:100%。LC/MS(ESI+)calcd for C13H13ClN2O+(M+H+)m/z,249.1;found249.1
3.化合物2-氯-4-((2-(6-氯哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈的合成化合物4-(2-氮杂螺[3.3]庚-6-基氧基)-2-氯苄腈三氟醋酸盐(363mg,1.00mmol)、6-氯哒嗪-3-羧酸(316mg,2.00mmol)和HATU(570mg,1.50mmol)加入到10mL二氯甲烷,冰水浴,加入DIPEA(645mg,5.00mmol)。撤掉冰浴,室温搅拌过夜。加二氯甲烷和水萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物2-氯-4-((2-(6-氯哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈(222mg,0.57mmol),收率:57%。
LC/MS(ESI+)calcd for C18H14Cl2N4O2 +(M+H+)m/z,389.0;found389.0
4.化合物叔丁基4-((1-(6-(6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羰基)哒嗪-3-基)哌啶-4-基)甲基哌嗪-1-酰胺的合成
将化合物2-氯-4-((2-(6-氯哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈(222mg,0.57mmol)、4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(323mg,1.14mmol)和DIPEA(221mg,1.71mmol)溶于2mL DOX,110℃下搅拌过夜。反应液加入水并用乙酸乙酯萃取,有机相用饱和氯化铵溶液和饱和氯化钠溶液洗涤三次,无水硫酸钠干燥,过滤、浓缩,硅胶柱层析纯化。得到化合物叔丁基4-((1-(6-(6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羰基)哒嗪-3-基)哌啶-4-基)甲基哌嗪-1-酰胺(150mg,0.23mmol),收率:40%。
LC/MS(ESI+)calcd for C33H42ClN7O4 +(M+H+)m/z,636.3;found636.3.
5.化合物2-氯-4-((2-(6-(4-(4-哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈的合成
化合物叔丁基4-((1-(6-(6-(3-氯-4-氰基苯氧基)-2-氮杂螺[3.3]庚烷-2-羰基)哒嗪-3-基)哌啶-4-基)甲基哌嗪-1-酰胺(150mg,0.23mmol)和2mL三氟醋酸溶于1mL二氯甲烷中,室温搅拌1h后,TLC检测反应完全,浓缩反应液,得到白色固体化合物2-氯-4-((2-(6-(4-(4-哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈三氟醋酸盐,直接作为下一步反应的原料,收率:100%。
LC/MS(ESI+)calcd for C28H34ClN7O2 +(M+H+)m/z,536.2;found536.2.
6.化合物2-氯-4-((2-(6-(4-((4-(2-(2,6-二氧哌啶-3-基))-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈的合成
将化合物2-氯-4-((2-(6-(4-(4-哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈三氟醋酸盐(153mg,0.23mmol),2-(2,6-二氧代哌啶-3-基)-5-氟-1,3-二氧代异吲哚啉(64mg,0.23mmol)和DIPEA(149mg,1.15mmol)加入到2mLDMSO。加热至130℃,搅拌3h。冷却至室温,加水和乙酸乙酯萃取,有机层用饱溶液和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物2-氯-4-((2-(6-(4-((4-((2-(2,6-二氧哌啶-3-基))-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)-2-氮杂螺[3.3]庚-6-基)氧基)苄腈(40mg,0.50mmol),收率:22%。LC/MS(ESI+)calcd for C41H42ClN9O6 +(M+H+)m/z,792.3;found792.3.1H NMR(400MHz,DMSO-d6)δ11.93(s,2H),11.09(s,1H),7.89(d,J=4.4Hz,1H),7.73(dd,J=6.0,3.2Hz,1H),7.68(d,J=4.0Hz,1H),7.35(s,1H),7.32–7.23(m,3H),7.05–7.00(m,1H),5.07(dd,J=8.8,3.6Hz,1H),4.86-4.79(m,1H),4.68-4.55(m,2H),4.67(s,1H),4.56(s,1H),4.15(s,1H),4.07(s,1H),3.48-3.43(m,3H),3.05-2.95(m,2H),2.84-2.78(m,2H),2.71-2.66(m,1H),2.32-2.26(m,2H),2.25-2.18(m,2H),2.06-1.95(m,2H),1.91(s,2H),1.87-1.79(m,2H),1.26-1.21(m,2H),1.18-1.08(m,3H).
121:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚-2-基)甲基)哌啶-1-基)哒嗪-3-酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-酰胺的合成
化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-酰胺(300mg,0.77mmol)、4-羟甲基哌啶(177mg,1.54mmol)和DIPEA(298mg,2.31mmol)溶于5mL DOX中搅拌,升温至110℃后搅拌过夜。反应液加入水并用乙酸乙酯萃取,有机相用饱和氯化铵溶液和饱和食盐水洗涤三次,无水硫酸钠干燥,浓缩,硅胶柱层析纯化,得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-酰胺(320mg,0.68mmol),收率:88%。
LC/MS(ESI+)calcd for C23H24ClN5O3 +(M+H+)m/z,470.2;found407.2.
2.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基))环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-酰胺(320mg,0.68mmol)和戴斯-马丁氧化剂(377mg,0.89mmol)溶于5mL二氯甲烷中,室温搅拌1.5h,TLC点板确定反应完全。过滤反应液,滤液用饱和亚硫酸氢钠溶液洗涤2次,用饱和碳酸钠溶液洗涤1次,用饱和食盐水洗涤1次,无水硫酸钠干燥,浓缩即得N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-酰胺(300mg,0.64mmol),收率94%。
LC/MS(ESI+)calcd for C24H36ClN5O3 +(M+H+)m/z,468.2;found468.2.
3.化合物叔丁基-6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-酰胺(300mg,0.64mmol)和2,6-二氮杂螺并[3.3]庚烷-2-羧酸叔丁酯(255mg,1.28mmol)溶于5mL 1,2-二氯乙烷之中,在两滴冰醋酸的催化之下,室温搅拌半个小时后,加入三乙酰氧基硼氢化钠(206mg,0.96mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到白色固体化合物叔丁基6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺(150mg,0.23mmol),收率:36%。
LC/MS(ESI+)calcd for C34H44ClN4O5 +(M+H+)m/z,650.3;found650.3.
4.化合物6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺的合成
化合物叔丁基-6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺(50mg,0.77mmol)和2mL三氟醋酸溶于1mL二氯甲烷中,室温搅拌1h后,TLC检测反应完全,浓缩反应液,得到白色固体化合物6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺三氟醋酸盐,直接作为下一步反应的原料,收率:100%。
LC/MS(ESI+)calcd for C29H36ClN7O2 +(M+H+)m/z,550.3;found550.3.
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚-2-基)甲基)哌啶-1-基)哒嗪-3-酰胺的合成
将化合物6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺三氟醋酸盐(51mg,0.077mmol),2-(2,6-二氧代哌啶-3-基)-5-氟-1,3-二氧代异吲哚啉(22mg,0.077mmol)和DIPEA(50mg,0.39mmol)加入到1mL DMSO。加热至130℃,搅拌4h。冷却至室温,加水和乙酸乙酯萃取,有机层用饱溶液和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚-2-基)甲基)哌啶-1-基)哒嗪-3-酰胺(20mg,0.025mmol),收率:32%。
LC/MS(ESI+)calcd for C42H44ClN9O6 +(M+H+)m/z,806.2;found805.8.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.61(d,J=4.0Hz,1H),7.86(d,J=4.4Hz,1H),7.80(d,J=4.4Hz,1H),7.65(d,J=4.4Hz,1H),7.39(s,1H),7.34(d,J=4.4Hz,1H),7.14(d,J=4.4Hz,1H),6.80(s,1H),6.66(d,J=4.0Hz,1H),5.10-5.03(m,1H),4.59-4.41(m,3H),4.16-4.09(m,3H),3.93-3.74(m,2H),3.20-3.14(m,1H),3.05-2.93(m,2H),2.91-2.81(m,1H),2.63-2.58(m,1H),2.54(s,3H),2.14-2.05(m,2H),2.03-1.99(m,1H),1.94-1.84(m,2H),1.84-1.72(m,2H),1.71-1.58(m,3H),1.57-1.45(m,2H),1.30-1.20(m,2H),1.20-1.03(m,3H).
122:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚-2-基)甲基)哌啶-1-基)哒嗪-3-酰胺的合成
LC/MS(ESI+)calcd for C42H43ClFN9O6 +(M+H+)m/z,824.3;found823.8.
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.60(d,J=4.0Hz,1H),7.85(d,J=4.4Hz,1H),7.79(d,J=4.4Hz,1H),7.60(d,J=4.4Hz,1H),7.34(s,1H),7.32(d,J=4.0Hz,1H),7.13(d,J=4.4Hz,1H),6.92(s,1H),5.14-4.99(m,1H),4.60-4.50(m,1H),4.50-4.39(m,2H),4.30-4.17(m,4H),4.08-3.96(m,1H),3.92-3.80(m,1H),3.31-3.23(m,4H),3.02-2.83(m,3H),2.63-2.57(m,1H),2.57-2.53(m,2H),2.31-2.20(m,2H),2.14-2.06(m,2H),2.05-1.96(m,2H),1.93-1.85(m,2H),1.81-1.71(m,2H),1.26-1.06(m,4H).
123:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚-2-基)甲基)哌啶-1-基)吡嗪-2-酰胺的合成
LC/MS(ESI+)calcd for C42H43ClFN9O6 +(M+H+)m/z,824.3;found824.2.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.58(s,1H),8.24(s,1H),8.08(d,J=4.0Hz,1H),7.86(d,J=4.4Hz,1H),7.61(d,J=5.6Hz,1H),7.38(s,1H),7.13(d,J=4.4Hz,1H),6.92(d,J=4.0Hz,1H),5.10-5.03(m,1H),4.56-4.48(m,1H),4.48-4.39(m,2H),4.28-4.21(m,3H),3.85-3.79(m,1H),3.31-3.29(m,2H),3.00-2.81(m,4H),2.71-2.67(m,1H),2.63-2.58(m,1H),2.21-2.14(m,1H),2.13-2.05(m,2H),2.05-1.97(m,2H),1.90-1.85(m,2H),1.80-1.71(m,2H),1.63-1.57(m,2H),1.53-1.48(m,2H),1.13-1.07(m,2H),0.89-0.80(m,3H).
124:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺的合成
1.叔丁基6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺的合成
化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-酰胺(200mg,0.51mmol),2,6-二氮杂螺并[3.3]庚烷-2-羧酸叔丁酯(202mg,1.02mmol)和DIPEA(198mg,1.53mmol)加入到5mL DOX中。加热到110℃,搅拌过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层再用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体化合物叔丁基6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺(200mg,0.36mmol),收率:71%。
LC/MS(ESI+)calcd for C23H35N2O5 +(M+H+)m/z,553.2;found553.2.
2.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺
化合物叔丁基6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-酰胺(200mg,0.36mmol)和2mL三氟醋酸溶于1mL二氯甲烷,室温搅拌1小时之后,TLC检测反应完全,浓缩后,加入饱和碳酸钠溶液调节pH值到10,加入二氯甲烷萃取。有机相用饱和食盐水洗涤,无水硫酸钠干燥,浓缩后即得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺(150mg,0.33mmol),收率:92%。LC/MS(ESI+)calcd for C22H33N2O5 +(M+H+)m/z,453.2;found453.2.
3.叔丁基4-((6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-基)哌啶-1-甲酸甲酯的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺(150mg,0.33mmol)、4-甲酰基哌啶-1-羧酸叔丁酯(141mg,0.66mmol)溶于3mL1,2-二氯乙烷之中,在两滴冰醋酸的催化之下,室温搅拌半个小时后,加入三乙酰氧基硼氢化钠(186mg.0.49mmol),室温搅拌过夜。反应液浓缩之后,加入水和二氯甲烷萃取,有机相用饱和氯化铵溶液和饱和食盐水洗涤,无水硫酸钠干燥,浓缩,硅胶柱层析纯化,得到化合物4-叔丁基4-((6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-基)哌啶-1-甲酸甲酯(100mg,0.15mmol),收率:46%。
LC/MS(ESI+)calcd for C35H46ClN4O5 +(M+H+)m/z,637.3;found637.2.
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(哌啶-4-基-甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺的合成
化合物4-叔丁基4-((6-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)-2,6-二氮杂螺[3.3]庚烷-2-基)哌啶-1-甲酸甲酯(100mg,0.15mmol)和2mL三氟醋酸溶于1mL二氯甲烷中,室温搅拌1h后,TLC检测反应完全,浓缩反应液,得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(哌啶-4-基-甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺三氟醋酸盐,直接作为下一步反应的原料,收率:100%。
LC/MS(ESI+)calcd for C30H38ClN4O3 +(M+H+)m/z,537.3;found537.3.
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)苯甲酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(哌啶-4-基-甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺三氟醋酸盐(97mg,0.15mmol),2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(44mg,0.15mmol)和DIPEA(97mg,0.75mmol)加入到3mLDMSO。加热至130℃,搅拌3h。冷却至室温,加水和乙酸乙酯萃取,有机相用饱和氯化铵溶液和饱和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)苯甲酰胺(18mg,0.022mmol),收率:15%。LC/MS(ESI+)calcd for C42H43ClN9O6 +(M+H+)m/z,824.3;found824.2.1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.64(d,J=4.0Hz,1H),7.83(d,J=4.4Hz,2H),7.61(d,J=6.4Hz,1H),7.50(d,J=1.2Hz,1H),7.36(d,J=4.8Hz,1H),7.17(dd,J=5.2,2.8Hz,1H),7.09(d,J=4.0Hz,1H),5.86-5.70(m,1H),5.10-5.03(m,1H),4.57-4.49(m,1H),3.90-3.83(m,3H),3.78-3.68(m,4H),3.62-3.58(m,2H),3.31(s,3H),2.98-2.80(m,2H),2.64-2.54(m,4H),2.41-2.31(m,3H),2.13-2.07(m,2H),2.05-1.97(m,2H),1.94-1.90(m,1H),1.89-1.87(m,1H),1.65-1.62(m,2H),1.25-1.23(m,2H).
125:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-基)吡嗪-2-酰胺的合成
LC/MS(ESI+)calcd for C42H43ClN9O6 +(M+H+)m/z,824.3;found824.2.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.59(s,1H),8.14(d,J=4.0Hz,1H),7.87–7.81(m,2H),7.73(d,J=5.6Hz,1H),7.46(m,J=4.0Hz,1H),7.39-7.36(m,1H),7.15-7.11(m,1H),5.14-5.07(m,1H),4.58-4.47(m,2H),4.30-4.23(m,4H),3.87-3.78(m,2H),3.66-3.56(m,4H),3.11-3.05(m,2H),2.96-2.83(m,5H),2.12-2.04(m,3H),1.89-1.84(m,2H),1.81-1.76(m,2H),1.62-1.57(m,2H),1.53-1.48(m,2H),1.24-1.23(m,3H),0.89-0.80(m,2H).
126:N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(4-(((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-酰胺的合成
1.化合物4-(((1r,4r)-4-氨基环己基)氧基)-2-溴苄腈的合成
化合物氢化钠(60%,3.50g,87.50mmol)溶于400mLDMF,在-10~0℃下搅拌10min后,加入(1r,4r)-4-羟基环己胺盐酸盐(4.17g,27.5mmol),在-10~0℃下搅拌30min后,缓慢滴加2-溴-4-氟苯腈(5.00g,25.00mmol)的DMF溶液,在-10~0℃下搅拌1h后,自然升温至室温后继续搅拌4h。反应液加入冰水混合物和乙酸乙酯萃取,有机层用饱和食盐水洗涤3次。有机相用1mol/L稀盐酸调节pH至1,用乙酸乙酯反萃取三次后,加入饱和氢氧化钠溶液调节pH至12,用乙酸乙酯萃取,有机相用饱和食盐水洗涤,无水硫酸钠干燥,浓缩之后,即得化合物4-(((1r,4r)-4-氨基环己基)氧基)-2-溴苄腈(5.20g,17.63mmol),收率:71%。
LC/MS(ESI+)calcd for C13H15BrN2O+(M+H+)m/z,295.0;found295.0.
2.化合物N-((1r,4r)-4-(3-溴-4-基苯氧基)环己基)-6-氯哒嗪-3-酰胺的合成
化合物4-(((1r,4r)-4-氨基环己基)氧基)-2-溴苄腈(1.00g,3.39mmol)、5-氯吡嗪-2-羧酸(805mg,5.08mmol)和HATU(1.67g,4.41mmol)加入到30mL二氯甲烷,冰水浴,加入DIPEA(1.13g,10.17mmol)。撤掉冰浴,室温搅拌过夜。加水和乙酸乙酯萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物N-((1r,4r)-4-(3-溴-4-基苯氧基)环己基)-6-氯哒嗪-3-酰胺(1.30g,2.98mmol),收率:88%。
LC/MS(ESI+)calcd for C18H16BrClN4O2 +(M+H+)m/z,435.0;found435.0.
3.叔丁基4-(6-(((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌嗪-1-酰胺
化合物N-((1r,4r)-4-(3-溴-4-基苯氧基)环己基)-6-氯哒嗪-3-酰胺(200mg,0.46mmol)、哌嗪-1-羧酸叔丁酯(172mg,0.92mmol)和DIPEA(178mg,1.38mmol)溶于3mLDOX中搅拌,升温至110℃后搅拌过夜。反应液加入水并用乙酸乙酯萃取,有机相用饱和氯化铵溶液和饱和食盐水洗涤三次,无水硫酸钠干燥,浓缩,硅胶柱层析纯化,得化合物叔丁基4-(6-(((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌嗪-1-酰胺(150mg,0.26mmol),收率:57%。LC/MS(ESI+)calcd for C27H33BrN6O4 +(M+H+)m/z,585.2;found585.2.
4.化合物N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-酰胺的合成
化合物叔丁基4-(6-(((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌嗪-1-酰胺(150mg,0.26mmol)溶于1mL二氯甲烷和2mL三氟乙酸,室温搅拌2h。溶剂旋干,加入饱和碳酸钠调节pH至12,使用二氯甲烷萃取,有机相用饱和食盐水,无水硫酸钠干燥,浓缩后即得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(哌啶-4-酰氧基)哌啶-1-基)苯甲酰胺(60mg,0.12mmol),收率:46%。
LC/MS(ESI+)calcd for C22H25BrN6O2 +(M+H+)m/z,485.1;found485.1.
5.N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(4-(((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-酰胺
化合物N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-酰胺(60mg,0.12mmol)和(1R,5S,6r)-3-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-3-氮杂双环[3.1.0]己烷-6-甲醛(48mg,0.12mmol)溶于1mL 1,2-二氯乙烷之中,在两滴冰醋酸的催化之下,室温搅拌半个小时后,加入三乙酰氧基硼氢化钠(27mg,0.12mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)6-(4-(((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]己烷-6-基)甲基)哌嗪-1-基)哒嗪-3-酰胺(16mg,0.019mmol),收率:16%。
LC/MS(ESI+)calcd for C41H41BrFN9O6 +(M+H+)m/z,854.2;found854.2.
1HNMR(400MHz,DMSO-d6)δ11.10(s,1H),8.64(d,J=4.0Hz,1H),7.84(d,J=4.0Hz,2H),7.61(d,J=6.4Hz,1H),7.52-7.48(m,1H),7.36(d,J=3.6Hz,1H),7.19-7.15(m,1H),7.08(d,J=3.6Hz,1H),5.10-5.04(m,1H),4.57-4.50(m,1H),3.90-3.81(m,3H),3.76-3.68(m,4H),3.63-3.58(m,2H),2.63-2.55(m,5H),2.37-2.33(m,2H),2.13-2.07(m,2H),1.95-1.85(m,3H),1.65-1.59(m,3H),1.27-1.20(m,4H),0.88-0.80(m,2H).
127:N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(4-(((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]己烷-6-基)甲基)哌嗪-1-基团)吡嗪-2-酰胺的合成
LC/MS(ESI+)calcd for C41H41BrFN9O6 +(M+H+)m/z,854.2;found854.3.
1HNMR(400MHz,DMSO-d6)δ11.10(s,1H),8.61(s,1H),8.27(s,1H),8.13(d,J=4.0Hz,1H),7.84(d,J=4.0Hz,1H),7.60(d,J=6.4Hz,1H),7.48(d,J=1.2Hz,1H),7.19–7.13(m,1H),7.08(d,J=4.0Hz,1H),4.56-4.46(m,5H),3.89-3.80(m,3H),3.76-3.65(m,4H),3.63-3.55(m,2H),2.94-2.81(m,1H),2.61-2.55(m,4H),2.41-2.31(m,2H),2.13-2.05(m,2H),1.99(s,2H),1.91-1.83(m,1H),1.68-1.58(m,3H),1.58-1.43(m,3H),1.27-1.21(m,1H),0.90-0.74(m,2H).
128:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-(((((1R,5S,6s)-3-(2-(2,6-二氧杂哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]己烷-6-基)甲基)哌嗪-1-基)嘧啶-5-酰胺
LC/MS(ESI+)calcd for C41H41BrFN9O6 +(M+H+)m/z,810found,810.3.
1HNMR(400MHz,DMSO-d6)δ11.11(s,1H),8.76(s,2H),8.15(d,J=3.6Hz,1H),7.86(d,J=4.4Hz,1H),7.60(d,J=6.4Hz,1H),7.39(s,1H),7.14(d,J=3.6Hz,1H),7.07(d,J=4.0Hz,1H),5.78-5.74(m,1H),5.10-5.02(m,1H),4.58-4.50(m,1H),3.86-3.80(m,5H),3.61-3.56(m,2H),2.92-2.81(m,1H),2.63-2.57(m,1H),2.35-2.29(m,2H),2.13-2.07(m,2H),2.03-1.98(m,1H),1.95-1.87(m,4H),1.62-1.58(m,2H),1.54-1.45(m,4H),1.27-1.19(m,3H),0.86-0.79(m,2H).
129:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-羟基-2-氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺
化合物6-羟基-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯(300mg,1.40mmol)和30mL三氟醋酸溶于15mL二氯甲烷,室温搅拌2h后,TLC检测反应完全,浓缩之后,加入6-氯-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-羧酰胺(200mg,0.51mmol)和DIPEA(330mg,2.55mmol)溶于5mL DOX中,升温至110℃搅拌过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-羟基-2-氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺(220mg,0.47mmol),收率:92%。
LC/MS(ESI+)calcd for C23H35N2O5 +(M+H+)m/z,468.2;found468.2.
2.N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-氧代-2-氮杂螺[3.3]庚-2-基)哒嗪-3-酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-羟基-2-氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺(220mg,0.41mmol)和戴斯-马丁氧化剂(225g,0.53mmol)溶于4mL二氯甲烷中,室温搅拌1h,TLC检测反应完全。反应液过滤之后,滤液用饱和亚硫酸氢钠溶液洗涤2次,用饱和碳酸钠洗涤1次和饱和食盐水洗涤1次,无水硫酸钠干燥,过滤,浓缩即得N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-氧代-2-氮杂螺[3.3]庚-2-基)哒嗪-3-酰胺(210mg,0.45mmol),收率:96%。
LC/MS(ESI+)calcd for C22H33N2O5 +(M+H+)m/z,466.2;found466.2.
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-2-氮杂螺[3.3]庚-2-基)哒嗪-3-酰胺的合成
化合物N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-氧代-2-氮杂螺[3.3]庚-2-基)哒嗪-3-酰胺(50mg,0.11mmol)和2-(2,6-二氧杂哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚啉-1,3-二酮(40mg,0.11mmol)溶于1mL1,2-二氯乙烷之中,加入两滴冰醋酸,室温搅拌半个小时之后,加入三乙酰氧基硼氢化钠(23mg,0.11mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-基)哒嗪-3-酰胺(22mg,0.027mmol),收率:25%。LC/MS(ESI+)calcdfor C41H41ClN9O6 +(M+H+)m/z,810.3;found810.3.1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),8.56(d,J=4.0Hz,1H),7.86(d,J=4.4Hz,1H),7.82(d,J=4.8Hz,1H),7.74(d,J=6.0Hz,1H),7.46(d,J=3.6Hz,1H),7.39(d,J=1.2Hz,1H),7.16-7.10(m,1H),6.83(d,J=4.8Hz,1H),5.76(s,1H),5.12(dd,J=9.2,4.0Hz,1H),4.58-4.49(m,1H),4.18(s,1H),4.06(s,1H),3.89-3.81(m,1H),3.29-3.21(m,4H),2.93-2.84(m,1H),2.47-2.37(m,5H),2.14-2.06(m,4H),1.93-1.86(m,2H),1.67-1.60(m,2H),1.56-1.47(m,3H),1.28-1.20(m,3H),0.89-0.78(m,1H).
130:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(6-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-基)吡嗪-2-酰胺的合成
LC/MS(ESI+)calcd for C41H41ClN9O6 +(M+H+)m/z,810.3;found810.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.57(s,1H),8.13(d,J=4.0Hz,1H),7.86(d,J=4.0Hz,1H),7.77-7.72(m,2H),7.46(d,J=4.4Hz,1H),7.38(d,J=1.2Hz,1H),7.13(dd,J=5.6,3.2Hz,2H),5.15-5.08(m,2H),4.55-4.47(m,1H),4.21-4.16(m,2H),4.09-4.06(m,2H),4.05-4.00(m,1H),3.86-3.77(m,1H),3.49-3.40(m,1H),3.27-3.22(m,4H),2.94-2.83(m,2H),2.76-2.66(m,2H),2.64-2.60(m,1H),2.59-2.56(m,1H),2.47-2.40(m,4H),2.13-2.03(m,2H),1.89-1.83(m,2H),1.62-1.49(m,2H).
131:N-((1r,4r)-4-(4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺合成
LC/MS(ESI+)calcd for C41H41ClN9O6 +(M+H+)m/z,800.4;found800.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.59(d,J=4.4Hz,1H),7.84(d,J=4.8Hz,1H),7.84(d,J=4.4Hz,2H),7.64(d,J=4.4Hz,1H),7.33(d,J=4.8Hz,1H),7.13(d,J=4.4Hz,2H),6.78(s,1H),6.65(d,J=4.0Hz,1H),5.05(dd,J=9.2,3.6Hz,1H),4.49-4.45(m,3H),3.85(m,1H),3.75(s,4H),3.35(s,5H),3.17(s,1H),3.00(t,J=8.0Hz,1H),2.91-2.85(m,1H),2.36-2.36(m,3H),2.16-2.10(m,4H),2.00-1.99(m,1H),1.77-1.77(m,5H),1.65-1.62(m,2H),1.55-1.46(m,2H),1.23-1.23(m,3H),1.14-1.03(m,3H).
132:N-((1r,4r)-4-(4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧杂哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺合成
LC/MS(ESI+)calcd for C41H41ClN9O6 +(M+H+)m/z818.4;found818.3.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.58(d,J=4.4Hz,1H),7.80(d,J=4.8Hz,1H),7.74(d,J=4.4Hz,2H),7.60(d,J=6.0Hz,1H),7.33(d,J=4.8Hz,1H),7.13(d,J=4.4Hz,2H),6.89(d,J=2.4Hz,1H),5.06(dd,J=9.2,4.0Hz,1H),4.51-4.46(m,3H),3.87(m,1H),3.76(s,4H),3.38(s,5H),3.16(s,1H),3.02(t,J=8.0Hz,1H),2.93-2.86(m,1H),2.38-2.35(m,3H),2.15-2.08(m,4H),2.03-1.99(m,1H),1.77-1.77(m,5H),1.66-1.61(m,2H),1.57-1.49(m,2H),1.25-1.25(m,3H),1.16-1.03(m,3H).
133:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺合成
1.化合物6-氯-N-(((1r,4r)-4-(3,4-(二氰基苯氧基)环己基)-N-氘代甲基哒嗪-3-酰胺的合成
化合物钠氢(60%)(27mg,0.68mmol)溶于2mLDMF,在-10~0℃下搅拌10min后,加入6-氯-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-羧酰胺(200mg,0.51mmol),在-10~0℃下搅拌30min后,缓慢滴加2-氯-4-氟苯腈(103mg,0.71mmol),在-10~0℃下搅拌1h后,自然升温至室温后继续搅拌2h。反应液加入水和乙酸乙酯萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到黄色油状化合物6-氯-N-(((1r,4r)-4-(3,4-(二氰基苯氧基)环己基)-N-氘代甲基哒嗪-3-酰胺(210mg,0.53mmol),收率:75%。
LC/MS(ESI+)calcd for C23H35N2O5 +(M+H+)m/z,399.2;found399.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-羟甲基哌啶-1-基)-N-氘代甲基哒嗪-3-羧酰胺的合成
化合物6-氯-N-(((1r,4r)-4-(3,4-(二氰基苯氧基)环己基)-N-氘代甲基哒嗪-3-酰胺(210mg,0.53mmol)和哌啶-4-基甲醇(92mg,0.80mmol)和DIPEA(205mg,1.59mmol)溶于5mL DOX中,升温至110℃搅拌过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层用饱和氯化铵溶液和饱和食盐水洗涤3次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-羟甲基哌啶-1-基)-N-氘代甲基哒嗪-3-酰胺(220mg,0.45mmol),收率:85%。
LC/MS(ESI+)calcd for C22H33N2O5 +(M+H+)m/z,487.2;found487.2.
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-羟甲基哌啶-1-基)-N-氘代甲基哒嗪-3-酰胺(220mg,0.45mmol)和戴斯-马丁氧化剂(246mg,0.58mmol)溶于4mL二氯甲烷中,室温搅拌1h,TLC检测反应完全。反应液过滤之后,滤液用饱和亚硫酸氢钠溶液洗涤2次,用饱和碳酸钠洗涤1次和饱和食盐水洗涤1次,无水硫酸钠干燥,过滤,浓缩即得N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺(200mg,0.45mmol),收率:87%。
LC/MS(ESI+)calcd for C35H46ClN4O5 +(M+H+)m/z,485.2;found485.2.
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺(50mg,0.10mmol)和2-(2,6-二氧杂哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚啉-1,3-二酮(36mg,0.10mmol)溶于1mL 1,2-二氯乙烷之中,加入两滴冰醋酸,室温搅拌半个小时之后,加入三乙酰氧基硼氢化钠(21mg,0.10mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基-哒嗪-3-酰胺(12mg,0.014mmol),收率:14%。LC/MS(ESI+)calcd forC42H42D3ClN9O6 +(M+H+)m/z,829.3;found829.3.
1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),7.87(t.J=8.8Hz,1H),7.74(d,J=6.0Hz,1H),7.51(d,J=4.8Hz,1H),7.46(d,J=3.6Hz,1H),7.41-7.30(m,2H),7.16-7.04(m,1H),5.76(s,1H),5.11(dd,J=9.2,4.0Hz,1H),4.61-4.32(m,4H),3.34(s,5H),3.33-3.30(m,1H),3.26(s,3H),3.02-2.84(m,3H),2.57-2.52(m,3H),2.27-2.19(m,2H),2.19-2.11(m,1H),2.10-2.01(m,2H),1.88-1.81(m,4H),1.77-1.69(m,1H),1.62-1.51(m,1H),1.38-1.20(m,3H),1.17-1.10(m,2H),0.92-0.71(m,2H).
134:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2-氮杂螺[3.3]庚烷-6-基)哌嗪-1-基)哒嗪-3-酰胺的合成
1.化合物叔丁基4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-酰胺的合成
化合物5-氯-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-酰胺(200mg,0.51mmol)、哌嗪-1-羧酸叔丁酯(172mg,0.92mmol)和DIPEA(178mg,1.38mmol)溶于5mL DOX中搅拌,升温至110℃后搅拌过夜。反应液加入水并用乙酸乙酯萃取,有机相用饱和氯化铵溶液和饱和食盐水洗涤三次,无水硫酸钠干燥,浓缩,硅胶柱层析纯化,得化合物叔丁基4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-酰胺(120mg,0.22mmol)。收率:43%。LC/MS(ESI+)calcd for C23H35N2O5 +(M+H+)m/z,419.2;found419.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-酰胺的合成
化合物叔丁基4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-酰胺(120mg,0.22mmol)和4mL三氟醋酸溶于2mL二氯甲烷,室温搅拌1h,TLC检测反应完全后,浓缩。浓缩后的剩余物加入饱和碳酸钠溶液调节pH至12,加入二氯甲烷萃取,有机相用饱和食盐水洗涤2次,加入无水硫酸钠干燥,过滤,浓缩即得N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-酰胺(80mg,0.18mmol),收率:82%。
LC/MS(ESI+)calcd for C22H33N2O5 +(M+H+)m/z,441.2;found441.2.
3.化合物叔丁基6-(4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-酰胺(80mg,0.18mmol)和6-氧代-2-氮杂螺[3.3]庚烷-2-羧酸叔丁酯(76mg,0.36mmol)溶于1mL1,2-二氯乙烷之中,加入两滴冰醋酸,室温搅拌半个小时之后,加入三乙酰氧基硼氢化钠(57mg,0.27mmol)后继续室温搅拌过夜。将反应液浓缩之后,加入水并二氯甲烷萃取,有机层用饱和氯化铵和食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物叔丁基6-(4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯(100mg,0.16mmol),收率:89%。
LC/MS(ESI+)calcd for C35H46ClN4O5 +(M+H+)m/z,636.3;found636.6.
4.化合物6-(4-(6-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯的合成
化合物叔丁基6-(4-(5-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)吡嗪-2-基)哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯(50mg,0.080mmol)和2mL三氟醋酸溶于1mL二氯甲烷,室温搅拌1h,TLC检测反应完全后,浓缩即得6-(4-(6-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯三氟醋酸盐(50mg,0.077mmol),收率:96%。LC/MS(ESI+)calcd for C30H38ClN4O3 +(M+H+)m/z,536.2;found536.2.
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)-2-氮杂螺[3.3]庚烷-6-基)哌嗪-1-基)吡嗪-2-酰胺的合成
化合物6-(4-(6-((((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基哌嗪-1-基)-2-氮杂螺[3.3]庚烷-2-羧酸酯三氟醋酸盐(50mg,0.077mmol),2-(2,6-二氧杂哌啶-3-基)-5,6-二氟异吲哚啉-1,3-二酮(23mg,0.077mmol)和DIPEA(50mg,0.39mmol)加入到1mL DMSO。加热至130℃,搅拌3h。冷却至室温,加水和乙酸乙酯萃取,有机层用0.5N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)-2-氮杂螺[3.3]庚烷-6-基)哌嗪-1-基)吡嗪-2-酰胺(12mg,0.015mmol),收率:19%。
LC/MS(ESI+)calcd for C41H41ClFN9O6 +(M+H+)m/z,810.3;found809.7.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.76(s,2H),8.14(d,J=3.6Hz,1H),7.86(d,J=4.4Hz,1H),7.59(d,J=5.6Hz,1H),7.39(d,J=1.2Hz,1H),7.14(dd,J=5.6,1.2Hz,1H),6.88(d,J=4.0Hz,1H),5.06(dd,J=9.2,3.6Hz,1H),4.60-4.50(m,1H),4.20(s,2H),4.09(s,2H),3.87–3.73(m,5H),2.94–2.81(m,1H),2.69-2.58(m,2H),2.58-2.53(m,1H),2.38-2.25(m,6H),2.13-1.96(m,6H),1.94-1.88(m,2H),1.54-1.45(m,4H).
135:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-2-羧酰胺
第一步:将2-(2,6-二氧哌啶-3-基)-5-氟异吲哚啉-1,3-二酮(553mg,2.0mmol),2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯(453mg,2.0mmol)和二异丙基乙胺(1.3g,10.0mmol)加入到10mL DMSO中。加热至135℃搅拌反应过夜。冷却至室温,加入乙酸乙酯和水萃取,有机相用用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物叔丁基2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬烷-7-羧酸酯(530mg)。收率:55%。LC/MS(ESI+)calcd for C25H30N4O6([M+H]+)m/z 482.22;found 483.1。
第二步:将化合物叔丁基2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬烷-7-羧酸酯(530mg,1.09mmol)溶于10mL二氯甲烷,并加入4mL三氟乙酸,室温搅拌反应3h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(342mg)。收率:82%。LC/MS(ESI+)calcd for C20H20N4O4([M+H]+)m/z 382.16;found 383.1。
第三步:将NaH(360mg,9.0mmol)加入到5mL DMF中,后将体系置于冰水浴中降温冷却搅拌,随后加入(1r,4r)-4-氨基环己烷-1-醇(455mg,3.0mmol),搅拌10min后,加入2-氯-4-氟苯甲腈(460mg,3mmol)。自然回温搅拌反应过夜。TLC监测原料消耗完毕,加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,无水硫酸钠干燥,旋干,Pre-TLC分离纯化。得化合物4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(279mg)。收率:62%。LC/MS(ESI+)calcdfor C13H15ClN2O([M+H]+)m/z 250.73;found 251.1。
第四步:将化合物5-氯嘧啶-2-羧酸(79mg,0.50mmol)和HATU(304mg,0.80mmol)加入到5mL二氯甲烷中,后将体系置于冰水浴中降温冷却搅拌,10min后加入二异丙基乙胺(194mg,1.50mmol),随后加入4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(150mg,0.50mmol)。完毕,撤去冰浴,室温搅拌反应过夜。TLC监测原料消耗完毕,加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,无水硫酸钠干燥,旋干,Pre-TLC分离纯化。得化合物5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)嘧啶-2-甲酰胺(185mg)。收率:92%。LC/MS(ESI+)calcd for C18H16Cl2N4O2([M+H]+)m/z 391.25;found 391.1。
第五步:将5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)嘧啶-2-甲酰胺(185mg,0.46mmol),哌啶-4-基甲醇(260mg,0.92mmol)和二异丙基乙胺(297mg,2.30mmol)加入到8mL二氧六环中。加热至115℃,回流搅拌反应过夜。冷却至室温,加入乙酸乙酯和水萃取,有机相用0.05N HCl洗涤,再用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)嘧啶-2-甲酰胺(109mg)。收率:37%。LC/MS(ESI+)calcd for C24H28ClN5O3([M+H]+)m/z 469.97;found 470.1。
第六步:将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)嘧啶-2-甲酰胺(50mg,0.11mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯-马丁氧化剂(72mg,0.17mmol),完毕,任体系自然升温至室温搅拌反应。2h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰哌啶-1-基)嘧啶-2-甲酰胺(30mg)。收率:58%。LC/MS(ESI+)calcd for C24H26ClN5O3([M+H]+)m/z 467.95;found468.1。
第七步:称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰哌啶-1-基)嘧啶-2-甲酰胺(30mg,0.06mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(24mg,0.06mmol),在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(38mg,0.18mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(21mg)。收率:39%。1H NMR(400MHz,DMSO-d6)δ8.51(s,2H),8.33(d,J=8.1Hz,1H),7.86(d,J=8.7Hz,1H),7.63(d,J=8.1Hz,1H),7.38(s,1H),7.13(d,J=8.4Hz,1H),6.78(s,1H),6.65(d,J=8.2Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.53(s,1H),3.97(d,J=11.3Hz,3H),2.87(t,J=11.9Hz,3H),2.60(s,2H),2.32(s,4H),2.12(s,4H),1.99(d,J=5.1Hz,1H),1.88(s,2H),1.79(s,2H),1.67-1.42(m,6H),1.23(s,4H),1.16(d,J=10.9Hz,3H),0.84(d,J=7.3Hz,2H)。LC/MS(ESI+)calcd for C44H48ClN9O6([M+H]+)m/z 834.38;found834.3。
136:N-((1r,4r)-4-((6-氰基-5-(三氟甲基)吡啶-3-基)氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C44H48F3N10O6([M+H]+)m/z 869。
137:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(3-((5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)氮杂苷-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.53(d,J=8.1Hz,1H),7.85(d,J=8.8Hz,1H),7.79(d,J=9.2Hz,1H),7.66-7.61(m,1H),7.38(d,J=2.4Hz,1H),7.17-7.11(m,2H),6.83(d,J=9.3Hz,1H),5.08(dd,J=12.9,5.4Hz,1H),4.53(s,1H),4.20(t,J=8.3Hz,2H),4.10(s,3H),3.85(s,1H),3.78(dd,J=8.4,5.5Hz,2H),3.70(s,2H),2.87(d,J=13.9Hz,3H),2.70(d,J=7.4Hz,2H),2.67-2.59(m,2H),2.09(d,J=11.3Hz,2H),2.05-1.99(m,1H),1.89(s,3H),1.70-1.56(m,2H),1.52(t,J=11.2Hz,2H),1.23(s,2H).LC/MS(ESI+)calcd for C41H41ClFN9O6([M+H]+)m/z 810.28;found 810.3。
138:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(3-((5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)壬苷-1-基)吡嗪-2-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.55(d,J=1.3Hz,1H),8.10(d,J=8.3Hz,1H),7.86(d,J=8.8Hz,1H),7.77(d,J=1.3Hz,1H),7.64(d,J=12.5Hz,1H),7.37(d,J=2.4Hz,1H),7.17-7.09(m,2H),5.76(s,2H),5.08(dd,J=12.9,5.3Hz,1H),4.50(s,1H),4.20(t,J=8.5Hz,2H),3.80(dd,J=8.8,5.5Hz,3H),3.70(s,2H),2.87(d,J=13.5Hz,3H),2.70(d,J=7.3Hz,2H),2.67-2.58(m,3H),2.55(d,J=10.3Hz,2H),2.08(s,3H),1.85(s,2H),1.56(dt,J=27.5,11.6Hz,4H),1.23(s,2H),1.05(t,J=7.0Hz,1H).LC/MS(ESI+)calcd for C41H41ClFN9O6([M+H]+)m/z 810.28;found 810.2。
139:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氨基)哌啶-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.56(d,J=8.2Hz,1H),7.85(d,J=8.8Hz,1H),7.80(d,J=9.6Hz,1H),7.57(d,J=10.3Hz,1H),7.38(d,J=2.3Hz,1H),7.32(d,J=9.7Hz,1H),7.18(d,J=7.1Hz,1H),7.13(dd,J=8.7,2.4Hz,1H),6.58(d,J=5.8Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),4.54(s,1H),4.47(d,J=12.6Hz,2H),3.87(s,1H),3.54(s,1H),3.40(d,J=5.9Hz,1H),3.01(t,J=12.1Hz,2H),2.85(d,J=12.5Hz,3H),2.60(s,1H),2.55(d,J=6.0Hz,2H),2.17(d,J=6.9Hz,2H),2.08(d,J=10.3Hz,2H),1.94-1.77(m,7H),1.67-1.58(m,3H),1.58-1.48(m,3H),1.24(s,1H),1.11(d,J=11.0Hz,2H).LC/MS(ESI+)calcd for C42H45ClFN9O6([M+H]+)m/z 826.33;found 826.2。
140:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(3-((5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)氮杂环丁烷-1-基)嘧啶-5-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.71(s,2H),8.14(d,J=7.5Hz,1H),7.86(d,J=8.8Hz,1H),7.63(d,J=12.5Hz,1H),7.38(d,J=2.4Hz,1H),7.17-7.10(m,2H),5.76(s,1H),5.08(dd,J=12.9,5.4Hz,1H),4.54(s,1H),4.16(t,J=8.6Hz,2H),3.75(dd,J=9.2,5.5Hz,3H),3.69(d,J=8.2Hz,2H),2.87(d,J=11.6Hz,4H),2.67(d,J=7.4Hz,2H),2.62(d,J=6.3Hz,2H),2.09(s,2H),2.06-1.96(m,1H),1.88(s,3H),1.49(s,4H),1.23(s,1H).LC/MS(ESI+)calcd for C41H41ClFN9O6([M+H]+)m/z 810.28;found 809.8。
141:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((7-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[4.4]壬-2-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.58(s,1H),8.24(s,1H),8.08(d,J=8.3Hz,1H),7.87(d,J=8.8Hz,1H),7.61(d,J=12.6Hz,1H),7.38(d,J=2.3Hz,1H),7.13(dd,J=8.8,2.2Hz,1H),7.05(d,J=7.5Hz,1H),5.07(dd,J=12.9,5.3Hz,1H),4.52(s,1H),4.45(d,J=12.2Hz,2H),3.81(s,1H),3.62(s,2H),3.50(dd,J=25.3,8.1Hz,3H),3.17(s,1H),2.97(t,J=12.5Hz,2H),2.87(d,J=12.0Hz,1H),2.62(s,1H),2.56(d,J=5.6Hz,2H),2.42(d,J=9.2Hz,1H),2.28(d,J=6.8Hz,2H),2.09(d,J=10.7Hz,2H),1.98(ddd,J=19.7,12.2,6.3Hz,3H),1.83-1.71(m,4H),1.67-1.56(m,2H),1.55-1.45(m,2H),1.28(d,J=16.1Hz,1H),1.24(s,1H),1.10(d,J=12.2Hz,2H),0.86(s,1H).LC/MS(ESI+)calcd forC44H47ClFN9O6([M+H]+)m/z 852.37;found 851.8。
142:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-2,7-二氮杂螺环[4.4]壬-2-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,CDCl3)δ8.18(s,1H),7.99(d,J=9.5Hz,1H),7.90(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.42(d,J=12.2Hz,1H),7.04(d,J=7.6Hz,1H),7.02(d,J=2.2Hz,1H),6.99(d,J=9.6Hz,1H),6.87(dd,J=8.7,2.2Hz,1H),4.94(dd,J=12.1,5.4Hz,1H),4.53(d,J=13.3Hz,2H),4.32(d,J=15.1Hz,1H),4.06(s,1H),3.73-3.59(m,3H),3.55(d,J=8.8Hz,1H),3.07(t,J=12.4Hz,3H),2.94(s,1H),2.89(s,2H),2.86-2.80(m,2H),2.77(d,J=16.8Hz,2H),2.60(d,J=5.5Hz,2H),2.18(d,J=10.2Hz,4H),2.07(d,J=7.3Hz,1H),2.01(d,J=11.3Hz,5H),1.74-1.64(m,2H),1.54-1.44(m,2H),1.35(s,1H),1.30(s,1H),0.90(s,1H).LC/MS(ESI+)calcd for C44H47ClFN9O6([M+H]+)m/z 852.37;found 852.2。
143:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氨基)哌啶-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,CDCl3)δ7.99(d,J=9.5Hz,1H),7.90(d,J=8.2Hz,1H),7.64(d,J=8.3Hz,1H),7.58(d,J=8.8Hz,1H),7.02(d,J=2.4Hz,1H),7.01(s,1H),6.88(dd,J=8.8,2.3Hz,1H),6.77(d,J=6.5Hz,1H),4.99-4.92(m,1H),4.53(d,J=12.7Hz,2H),4.34(s,1H),4.09(s,1H),3.90(s,1H),3.69(s,1H),3.50-3.42(m,1H),3.06(t,J=12.6Hz,3H),2.92(d,J=13.2Hz,1H),2.80(dd,J=24.9,12.5Hz,2H),2.38(s,2H),2.27(d,J=14.7Hz,3H),1.96(s,4H),1.70(s,4H),1.56-1.41(m,3H),1.29(d,J=11.8Hz,5H),0.90(s,1H).LC/MS(ESI+)calcd for C42H46ClN9O6([M+H]+)m/z 808.34;found 808.3。
145:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氨基)哌啶-1-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,CDCl3)δ8.85(s,1H),8.19(s,1H),7.98(s,1H),7.58(d,J=8.7Hz,1H),7.43(dd,J=15.2,9.1Hz,2H),7.11(d,J=7.0Hz,1H),7.01(d,J=2.4Hz,1H),6.87(dd,J=8.7,2.4Hz,1H),4.95(dd,J=12.2,5.4Hz,1H),4.62(s,1H),4.48(d,J=13.3Hz,2H),4.34(d,J=9.8Hz,1H),4.05(s,1H),3.48-3.40(m,1H),3.02(d,J=12.6Hz,1H),2.99-2.88(m,3H),2.87-2.73(m,3H),2.31(d,J=6.9Hz,2H),2.21(d,J=11.5Hz,5H),1.94(d,J=11.9Hz,3H),1.68(dd,J=23.4,11.7Hz,4H),1.54-1.41(m,3H),1.27(t,J=13.9Hz,4H),0.90(s,1H).LC/MS(ESI+)calcd for C42H45ClFN9O6([M+H]+)m/z 826.33;found 826.2。
146:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氨基)哌啶-1-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,CDCl3)δ8.84(s,1H),7.97(s,1H),7.63(d,J=8.3Hz,1H),7.58(d,J=8.7Hz,1H),7.41(d,J=8.2Hz,1H),7.01(d,J=2.2Hz,2H),6.87(dd,J=8.8,2.3Hz,1H),6.77(d,J=8.5Hz,1H),4.96(dd,J=12.1,5.4Hz,1H),4.81(s,1H),4.49(d,J=13.0Hz,2H),4.31(d,J=10.3Hz,1H),4.06(d,J=8.2Hz,2H),3.68(s,1H),3.16(s,2H),3.01(t,J=12.2Hz,2H),2.91(d,J=14.5Hz,1H),2.84-2.77(m,2H),2.49(s,3H),2.02(s,6H),1.78-1.63(m,4H),1.49(dd,J=22.3,10.8Hz,4H),1.33(d,J=19.5Hz,3H),0.90(s,1H).LC/MS(ESI+)calcd for C42H46ClN9O6([M+H]+)m/z 808.34;found 808.3。
147:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氧基)哌啶-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
第一步:将2-(2,6-二氧哌啶-3-基)-5-羟基异吲哚啉-1,3-二酮(274mg,1.0mmol),4-羟基哌啶-1-羧酸叔丁酯(201mg,1.0mmol),DEAD(209mg,1.2mmol)和三苯基磷(314mg,1.2mmol)加入到10mL THF中。氮气保护下,室温搅拌反应过夜。加入乙酸乙酯和水萃取,有机相用用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物叔丁基4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)氧基)哌啶-1-羧酸叔丁酯(326mg)。收率:71%。LC/MS(ESI+)calcd for C23H27N3O7([M+H]+)m/z 457.18;found 458.1。
第二步:将化合物叔丁基4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氧基)哌啶-1-羧酸叔丁酯(326mg,0.71mmol)溶于10mL二氯甲烷,并加入4mL三氟乙酸,室温搅拌反应3h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物2-(2,6-二氧哌啶-3-基)-5-(哌啶-4-氧基)异吲哚-1,3-二酮(213mg)。收率:84%。LC/MS(ESI+)calcd for C18H19N3O5([M+H]+)m/z 357.136;found358.2。
第三步:称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰哌啶-1-基)哒嗪-3-甲酰胺(57mg,0.12mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧哌啶-3-基)-5-(哌啶-4-氧基)异吲哚-1,3-二酮(44mg,0.12mmol),在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(100mg,0.48mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氧基)哌啶-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(23mg)。收率:33%。1H NMR(400MHz,DMSO-d6)δ8.60(d,J=8.4Hz,1H),7.88–7.83(m,2H),7.80(d,J=11.3Hz,1H),7.46(s,1H),7.40(d,J=2.3Hz,1H),7.37(d,J=8.3Hz,1H),7.33(d,J=9.8Hz,1H),7.14(dd,J=8.8,2.3Hz,1H),5.12(dd,J=13.0,5.4Hz,1H),4.72(s,1H),4.48(d,J=13.3Hz,3H),3.86(s,2H),3.05-2.96(m,3H),2.67(s,2H),2.25(s,2H),2.17(d,J=6.9Hz,2H),2.09(s,3H),1.97(s,2H),1.88(s,3H),1.82(d,J=12.7Hz,2H),1.70-1.61(m,4H),1.51(d,J=11.4Hz,3H),1.10(d,J=12.8Hz,2H).LC/MS(ESI+)calcd for C42H45ClN8O7([M+H]+)m/z:809.32;found809.2。
148:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-2-甲酰胺
第一步:将化合物5-溴嘧啶-2-羧酸(0.80g,3.94mmol)和HATU(2.41g,6.22mmol)加入到30mL二氯甲烷中,后将体系置于冰水浴中降温冷却搅拌,10min后加入二异丙基乙胺(1.03g,7.97mmol),随后加入4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(1.01g,3.99mmol)。完毕,撤去冰浴,室温搅拌反应过夜。TLC监测原料消耗完毕,加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,无水硫酸钠干燥,旋干,Pre-TLC分离纯化。得化合物5-溴-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)嘧啶-2-甲酰胺(1.09g)。收率:63.5%。LC/MS(ESI+)calcd for C18H16BrClN4O2([M+H]+)m/z 434.01;found 435.1。
第二步:将5-溴-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)嘧啶-2-甲酰胺(435mg,1.0mmol),哌啶-4-基甲醇(116mg,1.0mmol),醋酸钯(15mg,0.05mmol),二异丙基乙胺(35mg,0.05mmol)和碳酸铯(652mg,2.0mmol)加入到10mL二氧六环中。氮气保护下加热至110℃,回流搅拌反应过夜。冷却至室温,加入乙酸乙酯和水萃取,有机相用0.05N HCl洗涤,再用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)嘧啶-2-甲酰胺(191mg)。收率:41%。LC/MS(ESI+)calcd for C24H28ClN5O3([M+H]+)m/z 469.97;found470.1。
第三步:将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)嘧啶-2-甲酰胺(50mg,0.11mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯-马丁氧化剂(72mg,0.17mmol),完毕,任体系自然升温至室温搅拌反应。2h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰哌啶-1-基)嘧啶-2-甲酰胺(30mg)。收率:58%。LC/MS(ESI+)calcd for C24H26ClN5O3([M+H]+)m/z 467.95;found468.1。
第四步:称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰哌啶-1-基)嘧啶-2-甲酰胺(30mg,0.06mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(24mg,0.06mmol),在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(38mg,0.18mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-2-甲酰胺(21mg)。收率:39%。1H NMR(400MHz,DMSO-d6)δ8.51(s,2H),8.32(d,J=8.1Hz,1H),7.87(d,J=8.6Hz,1H),7.67(d,J=8.0Hz,1H),7.60(d,J=10.7Hz,1H),7.38(s,1H),7.29(d,J=7.9Hz,1H),7.23(d,J=7.4Hz,1H),7.13(d,J=9.5Hz,1H),6.90(d,J=7.7Hz,1H),6.85(s,1H),5.07(s,1H),4.53(s,1H),3.96(s,4H),2.86(d,J=12.7Hz,5H),2.64(d,J=28.8Hz,2H),2.33(s,4H),2.12(s,5H),2.05-1.96(m,2H),1.87(s,3H),1.63-1.48(m,6H),1.20-1.11(m,4H),0.85(s,2H).LC/MS(ESI+)calcd for C44H47ClFN9O6([M+H]+)m/z 852.37;found 852.3。
149:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-2-羧酰胺
1H NMR(400MHz,DMSO-d6)δ8.51(s,2H),8.33(d,J=8.1Hz,1H),7.86(d,J=8.7Hz,1H),7.63(d,J=8.1Hz,1H),7.38(s,1H),7.13(d,J=8.4Hz,1H),6.78(s,1H),6.65(d,J=8.2Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.53(s,1H),3.97(d,J=11.3Hz,3H),2.87(t,J=11.9Hz,3H),2.60(s,2H),2.32(s,4H),2.12(s,4H),1.99(d,J=5.1Hz,1H),1.88(s,2H),1.79(s,2H),1.67-1.42(m,6H),1.23(s,4H),1.16(d,J=10.9Hz,3H),0.84(d,J=7.3Hz,2H).LC/MS(ESI+)calcd for C44H48ClN9O6([M+H]+)m/z 834.38;found 834.3。
150:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮螺环[3.5]壬-2-基)嘧啶-2-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.31(d,J=8.2Hz,1H),8.05(s,2H),7.86(d,J=8.8Hz,1H),7.71(d,J=11.4Hz,1H),7.43(d,J=7.4Hz,1H),7.38(d,J=2.3Hz,1H),7.13(dd,J=8.8,2.2Hz,1H),5.10(dd,J=12.7,5.2Hz,1H),4.52(s,1H),3.60(d,J=10.9Hz,5H),2.88(t,J=12.2Hz,3H),2.32(s,3H),2.15(d,J=6.5Hz,2H),2.06(d,J=29.5Hz,4H),1.87(s,2H),1.78(s,5H),1.65–1.46(m,5H),1.23(s,6H),0.84(d,J=7.1Hz,1H).LC/MS(ESI+)calcd for C44H47ClFN9O6([M+H]+)m/z 852.37;found 852.3。
151:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.61(d,J=8.3Hz,1H),8.07(d,J=8.4Hz,1H),7.80(d,J=9.5Hz,1H),7.63(d,J=8.3Hz,1H),7.49(s,1H),7.33(d,J=9.7Hz,1H),6.78(d,J=1.7Hz,1H),6.70-6.60(m,1H),5.05(dd,J=12.8,5.4Hz,1H),4.63(s,2H),4.47(d,J=12.7Hz,3H),3.87(d,J=8.3Hz,2H),3.75(s,4H),3.00(t,J=11.8Hz,2H),2.88(dd,J=15.4,10.0Hz,1H),2.32(s,3H),2.12(d,J=7.0Hz,4H),2.05-1.95(m,1H),1.89(s,3H),1.82(s,3H),1.65(dd,J=24.3,11.0Hz,3H),1.60-1.46(m,3H),1.29-1.17(m,2H),0.86(dd,J=18.3,7.5Hz,1H).LC/MS(ESI+)calcd for C45H48F3N9O6([M+H]+)m/z 867.93;found 868.4。
152:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ8.61(d,J=8.4Hz,1H),8.07(d,J=8.4Hz,1H),7.80(d,J=9.5Hz,1H),7.59(d,J=11.2Hz,1H),7.51(s,1H),7.33(d,J=9.8Hz,1H),6.90(d,J=7.7Hz,1H),5.06(dd,J=12.9,5.5Hz,1H),4.63(s,1H),4.47(d,J=12.3Hz,2H),3.00(t,J=11.6Hz,2H),2.87(t,J=13.0Hz,1H),2.58(d,J=17.8Hz,1H),2.30(s,3H),2.12(d,J=6.8Hz,4H),2.06-1.96(m,1H),1.90(d,J=10.2Hz,3H),1.77(s,5H),1.68(s,5H),1.59-1.46(m,3H),1.23(s,2H),1.06(dd,J=17.3,10.3Hz,3H),0.84(d,J=7.5Hz,1H).LC/MS(ESI+)calcd for C45H47F4N9O6([M+H]+)m/z 885.92;found 886.4。
153:N-((1r,4r)-4-(4-氰基-3-(甲氧基-d3)苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.3Hz,1H),7.81(t,J=8.5Hz,1H),7.68-7.56(m,2H),7.32(d,J=9.7Hz,1H),6.77(s,1H),6.74-6.69(m,2H),6.65(d,J=8.3Hz,1H),5.76(s,1H),5.05(dd,J=12.9,5.4Hz,1H),4.47(d,J=11.7Hz,3H),3.85(s,1H),3.16(s,1H),3.00(t,J=11.8Hz,2H),2.94-2.81(m,1H),2.63-2.53(m,2H),2.32(s,4H),2.12(d,J=6.9Hz,4H),2.04-1.97(m,1H),1.89(s,3H),1.76(s,6H),1.64(dd,J=23.7,11.0Hz,2H),1.56-1.47(m,2H),1.23(s,2H),1.09(dd,J=22.4,9.9Hz,2H),0.84(d,J=6.8Hz,1H).LC/MS(ESI+)calcd for C45H48D3N9O7([M+H]+)m/z 832.98;found 833.3。
154:N-((1r,4r)-4-(4-氰基-3-乙氧基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.3Hz,1H),7.80(d,J=9.6Hz,1H),7.66-7.56(m,2H),7.32(d,J=9.8Hz,1H),6.78(d,J=1.8Hz,1H),6.70(dd,J=4.5,2.4Hz,2H),6.65(dd,J=8.4,1.9Hz,1H),5.76(s,1H),5.05(dd,J=12.8,5.4Hz,1H),4.53-4.42(m,3H),4.17(q,J=7.0Hz,2H),3.85(d,J=8.2Hz,1H),3.75(s,4H),3.16(s,1H),3.00(t,J=11.8Hz,2H),2.94-2.81(m,1H),2.57(dd,J=18.9,5.0Hz,2H),2.32(s,3H),2.12(d,J=6.5Hz,4H),2.04-1.96(m,1H),1.89(s,3H),1.76(s,4H),1.63(dd,J=24.3,10.8Hz,2H),1.50(dd,J=23.3,10.4Hz,2H),1.35(t,J=7.0Hz,3H),1.23(s,1H),1.17-1.01(m,2H),0.84(d,J=6.9Hz,1H).LC/MS(ESI+)calcd for C46H53N9O7([M+H]+)m/z843.99;found 844.4。
155:N-((1r,4r)-4-(4-氰基-3-(2-甲氧基乙氧基)苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.59(d,J=8.1Hz,1H),7.80(d,J=9.5Hz,1H),7.62(dd,J=12.7,8.5Hz,2H),7.33(d,J=9.5Hz,1H),6.76(d,J=9.6Hz,2H),6.71(d,J=8.6Hz,1H),6.65(d,J=8.5Hz,1H),5.76(s,2H),5.05(dd,J=12.8,5.4Hz,1H),4.47(d,J=10.1Hz,3H),4.30-4.22(m,2H),3.85(s,1H),3.75(s,4H),3.71-3.65(m,2H),3.30(s,2H),3.17(d,J=5.3Hz,1H),3.00(t,J=11.7Hz,2H),2.86(d,J=10.9Hz,1H),2.59(s,2H),2.33(s,4H),2.12(s,4H),1.99(s,1H),1.89(d,J=9.7Hz,3H),1.79(d,J=21.6Hz,6H),1.63(d,J=12.9Hz,2H),1.50(d,J=12.8Hz,2H),1.17-1.01(m,3H),0.85(s,1H).LC/MS(ESI+)calcd for C47H55N9O8([M+H]+)m/z 874.01;found 875.0。
156:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-羧酰胺
第一步:将5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(185mg,0.46mmol),4-((六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-羧酸叔丁酯(260mg,0.92mmol)和二异丙基乙胺(297mg,2.30mmol)加入到8mL二氧六环中。加热至115℃,回流搅拌反应过夜。冷却至室温,加入乙酸乙酯和水萃取,有机相用0.05N HCl洗涤,再用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物叔丁基5-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸酯(109mg)。收率:37%。LC/MS(ESI+)calcd for C29H35ClN6O4([M+H]+)m/z 567.09;found 567.1。
第二步:将化合物叔丁基5-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸酯(200mg,0.41mmol)溶于5mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应3h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-甲酰胺(142mg)。收率:90%。LC/MS(ESI+)calcd for C24H27ClN6O2([M+H]+)m/z 466.97;found 467.1。
第三步:称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-甲酰胺(30mg,0.06mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL),室温搅拌溶解澄清。随后向体系中加入1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)哌啶-4-乙醛(24mg,0.06mmol),在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(38mg,0.18mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-羧酰胺(21mg),收率:39%。1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.59(d,J=1.0Hz,1H),8.07(d,J=8.2Hz,1H),7.91(s,1H),7.84(d,J=8.7Hz,1H),7.61(d,J=8.5Hz,1H),7.35(d,J=2.3Hz,1H),7.26(s,1H),7.18(d,J=8.7Hz,1H),7.11(dd,J=8.8,2.3Hz,1H),5.74(s,1H),5.03(dd,J=12.9,5.2Hz,1H),4.48(d,J=5.6Hz,1H),3.99(d,J=12.6Hz,2H),3.78(d,J=8.7Hz,3H),3.49(s,1H),3.39(d,J=11.1Hz,2H),2.91(s,2H),2.56(d,J=9.0Hz,3H),2.22(d,J=6.3Hz,2H),2.07(d,J=11.1Hz,2H),1.98(d,J=8.8Hz,2H),1.85(s,2H),1.74(d,J=11.0Hz,3H),1.59(d,J=12.9Hz,2H),1.55-1.43(m,3H),1.16-1.05(m,3H),0.82(d,J=7.0Hz,1H)。LC/MS(ESI+)calcd for C43H46ClN9O6([M+H]+)m/z 820.35;found 820.3。
157:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.60(s,1H),8.08(d,J=8.2Hz,1H),7.93(s,1H),7.86(d,J=8.8Hz,1H),7.69(d,J=11.7Hz,1H),7.41(d,J=7.2Hz,1H),7.37(s,1H),7.12(d,J=8.9Hz,1H),5.09(dd,J=12.7,5.3Hz,1H),4.51(s,1H),4.10(q,J=5.3Hz,2H),3.79(s,3H),3.57(d,J=11.7Hz,2H),3.41(d,J=11.6Hz,2H),3.17(d,J=5.2Hz,5H),2.94(s,2H),2.90-2.79(m,3H),2.59(d,J=9.0Hz,2H),2.28(d,J=6.7Hz,2H),2.07(s,2H),1.90-1.75(m,4H),1.60(d,J=12.4Hz,2H),1.56-1.47(m,2H),1.23(s,1H).LC/MS(ESI+)calcd for C43H45ClFN9O6([M+H]+)m/z 838.34;found 838.3。
158:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺
LC/MS(ESI+)Calcd for C43H46FClN10O6(M+H+)m/z,853.3;found 853.3。1H NMR(400MHz,CDCl3)δ8.81(s,1H),8.05(s,1H),7.57(dd,J=8.4,2.5Hz,2H),7.36(d,J=10.9Hz,1H),7.00(d,J=2.4Hz,1H),6.87–6.79(m,2H),4.91(dd,J=12.3,5.3Hz,2H),4.31(dd,J=12.2,8.3Hz,1H),4.10–4.01(m,1H),3.89(d,J=1.9Hz,4H),3.05(s,2H),2.94–2.65(m,4H),2.40(s,3H),2.23–2.15(m,5H),1.90–1.85(m,7H),1.69(d,J=12.4Hz,3H),1.46(d,J=12.7Hz,3H),1.25(s,3H).
159:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(5-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基(氮杂环丁烷-3-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)吡嗪-2-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.60(d,J=1.3Hz,1H),8.09(d,J=8.3Hz1H)7.93(d,J=1.2Hz,1H),7.86(d,J=8.8Hz,1H),7.57(d,J=11.2Hz1H),7.37(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),6.88(d,J=7.6Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),4.52(s,1H),4.24(t,J=8.4Hz,2H),3.85-3.79(m,3H),3.77(d,J=3.1Hz,1H),3.51(s,1H),3.41(d,J=9.9Hz,2H),2.94(s,2H),2.87(dd,J=15.8,10.1Hz,2H),2.69(s,1H),2.67(s,1H),2.60(s,2H),2.55(s,2H),2.17(t,J=8.0Hz,1H),2.09(d,J=11.6Hz,2H),2.00(d,J=5.5Hz,1H),1.91-1.86(m,2H),1.60(d,J=13.0Hz,1H),1.56-1.47(m,2H),1.30(s,1H),1.26(s,1H),0.84(d,J=7.1Hz,1H).LC/MS(ESI+)calcd for C41H41ClFN9O6([M+H]+)m/z 810.28;found 810.3。
160:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂环丁烷-3-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.57(d,J=8.1Hz,1H),7.84(dd,J=12.4,9.1Hz,2H),7.57(d,J=11.2Hz,1H),7.39(d,J=2.3Hz,1H),7.14(dd,J=8.8,2.3Hz,1H),7.00(d,J=9.4Hz,1H),6.88(d,J=7.7Hz,1H),5.76(s,1H),5.05(dd,J=12.8,5.3Hz,1H),4.54(s,1H),4.25(t,J=7.1Hz,2H),3.83(d,J=7.0Hz,3H),3.77(s,2H),3.41(d,J=11.3Hz,3H),2.96(s,2H),2.92-2.81(m,2H),2.62(d,J=6.3Hz,2H),2.10(d,J=11.1Hz,2H),2.00(d,J=5.6Hz,1H),1.91(d,J=8.3Hz,2H),1.78(s,1H),1.64(dd,J=23.8,10.9Hz,2H),1.57-1.46(m,2H),1.28(d,J=16.0Hz,1H),1.23(s,1H),0.88(d,J=23.7Hz,1H).LC/MS(ESI+)calcd for C41H41ClFN9O6([M+H]+)m/z 810.28;found 810.3。
161:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(5-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)壬二酸-3-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-吡唑)甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.25(d,J=9.3Hz,1H),7.91(d,J=8.7Hz,1H),7.85(d,J=9.4Hz,1H),7.57(d,J=11.2Hz,1H),7.25(d,J=2.4Hz,1H),7.03(dd,J=9.3,3.2Hz,2H),6.88(d,J=7.6Hz,1H),5.76(s,2H),5.05(dd,J=12.9,5.3Hz,1H),4.46(s,1H),4.25(t,J=7.2Hz,2H),4.01(d,J=9.2Hz,1H),3.85-3.74(m,4H),3.51(s,1H),3.44(d,J=7.4Hz,3H),2.97(s,2H),2.92-2.82(m,2H),2.67(dd,J=13.8,8.2Hz,4H),1.30(s,1H),1.26(s,1H),1.22(s,6H),1.14(s,6H).LC/MS(ESI+)calcd forC41H41ClFN9O6([M+H]+)m/z 838.34;found 838.2。
162:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[4.4]壬-2-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.55(d,J=8.1Hz,1H),7.84(dd,J=17.9,9.1Hz,2H),7.71(d,J=11.4Hz,1H),7.44(d,J=7.3Hz,1H),7.40(d,J=2.4Hz,1H),7.14(dd,J=8.8,2.4Hz,1H),6.95(d,J=9.5Hz,1H),5.10(dd,J=13.0,5.4Hz,1H),4.54(s,1H),3.85(s,1H),3.60(d,J=9.5Hz,3H),3.44(s,1H),3.17(d,J=5.1Hz,1H),2.94-2.82(m,3H),2.69-2.61(m,1H),2.55(d,J=10.1Hz,2H),2.43(d,J=9.2Hz,2H),2.32(d,J=7.4Hz,2H),2.10(s,2H),2.02(dd,J=13.1,6.4Hz,3H),1.90(d,J=13.6Hz,2H),1.82(d,J=6.3Hz,3H),1.71-1.58(m,3H),1.57-1.46(m,2H),1.29(s,1H),1.25(d,J=6.2Hz,3H),0.86(s,1H).LC/MS(ESI+)calcd for C44H47ClFN9O6([M+H]+)m/z 852.37;found 851.8。
163:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[4.4]壬-2-基)吡嗪-2-甲酰胺
1H NMR(400MHz,CDCl3)δ8.87(d,J=1.2Hz,1H),7.73(d,J=1.2Hz,1H),7.61-7.55(m,1H),7.47(d,J=11.0Hz,1H),7.43(d,J=8.2Hz,1H),7.39(d,J=7.3Hz,1H),7.01(d,J=2.4Hz,1H),6.87(dd,J=8.7,2.4Hz,1H),4.95(dd,J=12.3,5.3Hz,1H),4.40-4.26(m,1H),4.05(dd,J=11.7,7.2Hz,1H),3.72-3.58(m,6H),2.96-2.71(m,8H),2.54(d,J=6.9Hz,2H),2.24-2.13(m,5H),2.00-1.86(m,4H),1.71(dd,J=22.4,9.7Hz,3H),1.56-1.38(m,4H),1.33(d,J=19.3Hz,1H),1.27(s,1H),0.90(s,1H).LC/MS(ESI+)calcd forC44H47ClFN9O6([M+H]+)m/z 852.37;found 852.3。
164:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-2,7-二氮杂螺环[4.4]壬-2-基)哌啶-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.59(d,J=1.1Hz,1H),8.26(s,1H),8.09(d,J=8.2Hz,1H),7.87(d,J=8.8Hz,1H),7.59(d,J=12.6Hz,1H),7.38(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),7.04(d,J=7.6Hz,1H),5.76(s,1H),5.06(dd,J=12.9,5.4Hz,1H),4.53(d,J=9.9Hz,1H),4.27(d,J=9.7Hz,2H),3.83(d,J=7.3Hz,1H),3.62(s,2H),3.51(s,1H),3.46(d,J=10.4Hz,2H),3.21-3.13(m,5H),2.89(dd,J=22.4,8.2Hz,1H),2.72(d,J=6.3Hz,1H),2.66-2.58(m,2H),2.55(d,J=10.7Hz,1H),2.35(d,J=9.0Hz,1H),2.09(d,J=10.9Hz,2H),2.04-1.96(m,1H),1.90(d,J=11.8Hz,2H),1.87(s,2H),1.79(dd,J=14.7,7.1Hz,2H),1.67-1.56(m,2H),1.56-1.47(m,2H),1.47-1.38(m,2H),0.95(d,J=6.5Hz,1H).LC/MS(ESI+)calcd for C43H45ClFN9O6([M+H]+)m/z 838.34;found 837.8。
165:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[4.4]壬-2-基)哌啶-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.60(d,J=8.2Hz,1H),7.86(d,J=8.8Hz,1H),7.81(d,J=9.6Hz,1H),7.59(d,J=12.6Hz,1H),7.40(d,J=2.4Hz,1H),7.35(d,J=9.7Hz,1H),7.14(dd,J=8.8,2.4Hz,1H),7.05(d,J=7.5Hz,1H),5.06(dd,J=12.9,5.3Hz,1H),4.53(d,J=9.8Hz,1H),4.30(d,J=9.4Hz,2H),3.85(s,1H),3.62(s,2H),3.53-3.43(m,4H),3.22(s,1H),2.94-2.83(m,1H),2.72(d,J=6.1Hz,1H),2.63(t,J=9.8Hz,2H),2.55(d,J=11.0Hz,1H),2.35(d,J=13.6Hz,2H),2.11(d,J=9.8Hz,2H),2.01(dd,J=12.0,6.2Hz,1H),1.90(d,J=11.5Hz,3H),1.83(s,1H),1.78(dd,J=14.4,7.8Hz,2H),1.64(dd,J=23.9,10.6Hz,2H),1.56-1.49(m,2H),1.42(d,J=9.3Hz,2H),1.24(s,1H),0.95(d,J=6.6Hz,1H).LC/MS(ESI+)calcd for C43H45ClFN9O6([M+H]+)m/z 838.34;found 837.7。
166:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺环[4.4]壬-2-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.55(d,J=8.1Hz,1H),7.87(d,J=8.8Hz,1H),7.82(d,J=9.3Hz,1H),7.72(d,J=11.5Hz,1H),7.45(d,J=7.3Hz,1H),7.40(d,J=2.4Hz,1H),7.14(dd,J=8.8,2.4Hz,1H),6.95(d,J=9.6Hz,1H),5.11(dd,J=12.9,5.3Hz,1H),4.54(s,1H),3.85(s,2H),3.57(s,4H),3.17(s,1H),2.99-2.91(m,2H),2.90(s,1H),2.86(s,1H),2.75(d,J=10.1Hz,1H),2.70-2.59(m,3H),2.33(s,1H),2.26(s,1H),2.10(s,2H),2.01(d,J=13.9Hz,2H),1.91(d,J=14.5Hz,3H),1.81(s,2H),1.65(d,J=17.6Hz,2H),1.56-1.49(m,3H),1.25(d,J=6.2Hz,2H),0.86(s,1H).LC/MS(ESI+)calcdfor C43H45ClFN9O6([M+H]+)m/z 838.34;found 837.8。
167:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[4.4]壬-2-基)吡嗪-2-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.60(s,1H),8.08(d,J=8.2Hz,1H),7.96-7.82(m,2H),7.72(d,J=11.5Hz,1H),7.45(d,J=7.5Hz,1H),7.38(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.77(s,1H),5.11(dd,J=12.7,5.3Hz,1H),4.52(s,1H),3.84(s,1H),3.59(dd,J=25.2,19.3Hz,4H),3.53(d,J=10.9Hz,1H),3.44(d,J=10.8Hz,1H),3.00-2.91(m,2H),2.88(d,J=15.1Hz,1H),2.74(s,1H),2.69-2.61(m,2H),2.56(d,J=12.3Hz,2H),2.25(s,1H),2.09(d,J=11.8Hz,2H),1.97(dd,J=46.8,19.5Hz,5H),1.85(s,1H),1.81(s,1H),1.67-1.45(m,5H),1.32(d,J=15.0Hz,1H),1.27-1.22(m,1H),0.86(s,1H).LC/MS(ESI+)calcd for C43H45ClFN9O6([M+H]+)m/z 838.34;found 837.8。
168:N-((1r,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3S)-4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-3-甲基哌嗪-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,CDCl3)δ8.06(s,1H),8.00(d,J=9.5Hz,1H),7.87(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=12.5Hz,1H),7.03(d,J=7.3Hz,1H),6.99(dd,J=11.2,6.0Hz,2H),6.85(dd,J=8.7,2.3Hz,1H),4.91(dd,J=12.3,5.3Hz,1H),4.32(s,1H),4.18(d,J=12.7Hz,2H),4.05(d,J=8.1Hz,1H),3.87(d,J=10.2Hz,2H),3.59(d,J=9.3Hz,3H),3.41(s,1H),3.09(dd,J=25.2,13.8Hz,2H),2.90(d,J=13.5Hz,1H),2.84-2.78(m,1H),2.74(dd,J=12.2,5.5Hz,2H),2.60(s,1H),2.51(d,J=8.7Hz,1H),2.45-2.37(m,1H),2.24-2.10(m,6H),1.60(dd,J=8.2,3.7Hz,3H),1.52-1.37(m,4H),0.91(s,1H).LC/MS(ESI+)calcd for C42H43ClFN9O6([M+H]+)m/z 824.31;found 824.2。
169:N-((1r,4S)-4-(3-氯-4-氰基苯氧基)环己基)-5-((3S)-4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-3-甲基哌嗪-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,CDCl3)δ8.85(s,1H),8.05(s,1H),7.97(s,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=4.6Hz,1H),7.38(s,1H),7.03(d,J=7.4Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),4.91(dd,J=12.1,5.4Hz,1H),4.30(t,J=10.0Hz,1H),4.04(s,1H),3.89(s,2H),3.64-3.54(m,2H),2.94-2.69(m,4H),2.26-2.10(m,5H),1.65(d,J=13.5Hz,10H),1.53-1.39(m,4H),1.25(s,3H),0.88(s,1H).LC/MS(ESI+)calcd forC42H43ClFN9O6([M+H]+)m/z 824.31;found 824.3。
170:N-((1r,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((2S)-4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2-甲基哌嗪-1-基)哒嗪-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.65(d,J=8.2Hz,1H),7.85(t,J=9.5Hz,2H),7.62(s,1H),7.40(d,J=2.4Hz,1H),7.30(d,J=9.7Hz,1H),7.14(dd,J=8.8,2.3Hz,1H),7.09(d,J=7.5Hz,1H),5.05(d,J=5.3Hz,1H),4.66(s,1H),4.53(s,1H),4.22(d,J=11.7Hz,1H),3.92(dd,J=10.7,3.3Hz,1H),3.89-3.83(m,3H),3.70(d,J=10.2Hz,1H),3.61(d,J=9.1Hz,2H),3.21-3.12(m,2H),3.08(d,J=9.8Hz,1H),2.97-2.81(m,3H),2.60(s,1H),2.38-2.31(m,2H),2.26(d,J=7.9Hz,1H),2.10(d,J=11.0Hz,3H),2.05-1.96(m,2H),1.95-1.83(m,3H),1.51(dd,J=23.1,10.3Hz,3H),1.18(d,J=6.4Hz,1H),0.82(s,1H).LC/MS(ESI+)calcd for C42H43ClFN9O6([M+H]+)m/z 824.31;found 824.3。
171:N-((1r,4S)-4-(3-氯-4-氰基苯氧基)环己基)-5-((2S)-4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2-甲基哌嗪-1-基)吡嗪-2-羧酰胺
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.62(d,J=0.9Hz,1H),8.21(s,1H),8.12(d,J=8.3Hz,1H),7.86(d,J=8.8Hz,1H),7.61(d,J=12.8Hz,1H),7.38(d,J=2.4Hz,1H),7.16-7.04(m,2H),5.07(dd,J=12.8,5.4Hz,1H),4.65(s,1H),4.52(s,1H),4.27-4.18(m,1H),3.85(d,J=10.3Hz,3H),3.60(d,J=8.4Hz,2H),3.15(t,J=10.9Hz,2H),3.06(d,J=10.2Hz,1H),2.89(t,J=15.1Hz,2H),2.58(d,J=18.2Hz,2H),2.41-2.28(m,2H),2.24(d,J=7.6Hz,1H),2.08(d,J=7.5Hz,3H),2.05-1.97(m,2H),1.89(d,J=18.4Hz,2H),1.59(s,1H),1.57-1.43(m,3H),1.23(d,J=6.5Hz,3H),0.81(s,1H).LC/MS(ESI+)calcd forC42H43ClFN9O6([M+H]+)m/z:824.31;found 824.3。
172:2-氯-4-((1r,4r)-4-(2-((1-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)哌啶-4-基)氨基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
LC/MS(ESI+)calcd for C44H47ClN8O6([M+H]+)m/z819.36;found 819.3。
173:2-氯-4-((1r,4r)-4-(2-(4-((4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氨基)哌啶-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),7.88(d,J=8.8Hz,1H),7.71-7.68(m,1H),7.56(d,J=8.4Hz,1H),7.40(d,J=2.3Hz,1H),7.17-7.11(m,1H),7.04(d,J=7.1Hz,1H),6.98(s,1H),6.88(d,J=9.7Hz,1H),5.03(dd,J=12.7,5.1Hz,1H),4.57(s,1H),4.43(d,J=12.1Hz,2H),4.25(s,2H),4.07(s,1H),3.17(s,1H),2.93(t,J=12.3Hz,3H),2.82(d,J=10.3Hz,2H),2.58(d,J=16.2Hz,1H),2.16(d,J=6.4Hz,3H),2.12-2.03(m,2H),2.00(s,1H),1.92(d,J=10.9Hz,2H),1.85(s,2H),1.79(s,6H),1.56(s,2H),1.44(d,J=10.3Hz,2H),1.25(d,J=10.2Hz,1H),1.07(d,J=12.2Hz,2H).LC/MS(ESI+)calcd forC44H47ClN8O6([M+H]+)m/z 819.36;found 819.3。
174:2-氯-4-(((3aR,5r,6aS)-2-(5-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯甲腈
1.2-氯-4-(((3aR,5r,6aS)-2-(5-氯吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈
将化合物5-氯吡嗪-2-羧酸(79mg,0.50mmol)和HATU(304mg,0.80mmol)加入到5mL二氯甲烷中,后将体系置于冰水浴中降温冷却搅拌,10min后加入二异丙基乙胺(194mg,1.50mmol),随后加入2-氯-4-(((3aR,5r,6aS)-八氢环戊烷[c]吡咯-5-基)氧)苯腈盐酸盐(150mg,0.50mmol)。完毕,撤去冰浴,室温搅拌反应过夜。TLC监测原料消耗完毕,加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,无水硫酸钠干燥,旋干,Pre-TLC分离纯化。得化合物2-氯-4-(((3aR,5r,6aS)-2-(5-氯吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈(185mg)。收率:92%。LC/MS(ESI+)calcd for C19H16Cl2N4O2(M+H)+m/z,403.3;found,403.0.
2.叔丁基-4-((1-(5-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢环戊烷[c]吡咯-2-羰基)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯的合成
将2-氯-4-(((3aR,5r,6aS)-2-(5-氯吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈(185mg,0.46mmol),叔丁基-4-(哌啶-4-基甲基)哌嗪-1-羧酸酯(260mg,0.92mmol)和二异丙基乙胺(297mg,2.30mmol)加入到8mL二氧六环中。加热至115℃,回流搅拌反应过夜。冷却至室温,加入乙酸乙酯和水萃取,有机相用0.05N HCl洗涤,再用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品,Pre-TLC分离纯化。得到化合物叔丁基-4-((1-(5-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢环戊烷[c]吡咯-2-羰基)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯(109mg)。收率:37%。LC/MS(ESI+)calcd for C34H44ClN7O4(M+H)+m/z,650.2;found,650.2.
3.化合物2-氯-4-(((3aR,5r,6aS)-2-(5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈三氟乙酸盐的合成
将化合物叔丁基-4-((1-(5-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢环戊烷[c]吡咯-2-羰基)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯(109mg,0.17mmol)溶于6mL二氯甲烷和3mL三氟乙酸,室温搅拌反应2h。旋蒸除去溶剂,并用二氯甲烷旋带除去残留的三氟乙酸,得到化合物2-氯-4-(((3aR,5r,6aS)-2-(5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈三氟乙酸盐,不经进一步纯化,直接用于下步反应中。
LC/MS(ESI+)calcd for C31H37ClF3N7O4(M+H)+m/z,550.1;found,550.4.
4.2-氯-4-(((3aR,5r,6aS)-2-(5-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈
将2-氯-4-(((3aR,5r,6aS)-2-(5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈三氟乙酸盐,2-(2,6-二氧代哌啶-3-基)-5-氟异吲哚啉1,3-二酮(47mg,0.17mmol)和二异丙基乙胺(181mg,1.40mmol)加入到5mL二甲基亚砜中。后将体系移置于130℃的油浴中,搅拌反应过夜。冷却至室温,加乙酸乙酯和水萃取,有机层用0.05N HCl洗涤,再用饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂,Pre-TLC分离纯化。得到化合物2-氯-4-(((3aR,5r,6aS)-2-(5-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)吡嗪-2-羰基)八氢环戊烷[c]吡咯-5-基)氧)苯腈(22mg)。收率:16%。
LC/MS(ESI+)calcd for C31H37ClF3N7O4(M+H)+m/z,806.3;found,806.3.
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.46(s,1H),8.23(s,1H),7.85(d,J=8.7Hz,1H),7.68(d,J=8.4Hz,1H),7.34(s,1H),7.26(dd,J=8.2,1.6Hz,1H),7.20(d,J=2.1Hz,1H),7.02(dd,J=8.6,2.0Hz,1H),5.05(dt,J=13.2,7.8Hz,2H),4.43(d,J=11.6Hz,2H),3.91(dd,J=13.9,7.3Hz,1H),3.79(t,J=11.7Hz,1H),3.67(d,J=17.5Hz,2H),3.45(s,4H),3.01–2.92(m,2H),2.87(d,J=23.0Hz,1H),2.76(s,2H),2.59(d,J=31.6Hz,2H),2.33(s,3H),2.21(d,J=7.1Hz,2H),2.05–1.96(m,3H),1.83(d,J=15.2Hz,2H),1.76–1.58(m,4H),1.32(d,J=8.7Hz,1H),1.11(d,J=12.6Hz,1H).
175:2-氯-4-(((3aR,5s,6aS)-2-(6-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)八氢环戊[c]吡咯-5-基)氧)苯甲腈
LC/MS(ESI+)calcd for C42H44ClN9O6(M+H)+m/z,806.3;found,806.2.
1H NMR(400MHz,CDCl3)δ8.28(s,1H),7.87(d,J=9.4Hz,1H),7.70(d,J=8.4Hz,1H),7.56(d,J=8.9Hz,1H),7.29(s,1H),7.06(dd,J=7.0,1.1Hz,1H),7.02–6.87(m,2H),6.81(dd,J=8.1,1.9Hz,1H),4.94(s,3H),4.54(d,J=12.6Hz,2H),4.14(dd,J=42.5,8.5Hz,3H),3.98–3.79(m,1H),3.69(d,J=11.0Hz,1H),3.44(br,4H),3.17–2.67(m,7H),2.60(s,4H),2.36–2.19(m,3H),2.19–2.02(m,4H),1.95(d,J=11.0Hz,3H).
176:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺的合成
1.化合物(1R,5S,6r)-3-氮杂双环[3.1.0]己烷-6-基甲醇的合成
将化合物(1R,5S,6r)-叔丁基6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(50mg,0.23mmol)溶于3mL二氯甲烷,并加入1.5mL三氟乙酸,室温搅拌反应2h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸,得化合物(1R,5S,6r)-3-氮杂双环[3.1.0]己烷-6-基甲醇三氟乙酸盐,不经进一步纯化,直接用于下步反应中。
2.化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺的合成
向装有(1R,5S,6r)-3-杂双环[3.1.0]己烷-6-基甲醇三氟乙酸盐的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(82mg,0.21mmol),碳酸钾(95mg,0.69mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。4h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺(82mg,0.17mmol)。收率:76%。
LC/MS(ESI+)calcd for C24H26ClN5O3(M+H)+m/z,467.9;found,468.1.
3.化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺的合成
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺(80mg,0.17mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(144mg,0.34mmol),完毕,任体系自然升温至室温,并在室温搅拌反应过夜。翌日,TLC检测原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺(60mg)。收率:76%。
LC/MS(ESI+)calcd for C24H24ClN5O3(M+H)+m/z,465.9;found,466.1.
4.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺
称取N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺(25mg,0.05mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL)和甲醇(1mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧代哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚啉-1,3-二酮(18mg,0.05mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(32mg,0.15mmol),任体系在室温搅拌反应过夜。翌日,取样点板,TLC显示反应结束。停止搅拌,旋蒸除去溶剂后,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺(22mg)。收率:54%。LC/MS(ESI+)calcd for C41H41ClFN9O6(M+H)+m/z,810.3;found,810.3.
1H NMR(400MHz,CDCl3)δ8.84(d,J=1.2Hz,1H),8.56(br,1H),7.68(d,J=1.1Hz,1H),7.56(d,J=8.7Hz,1H),7.49(d,J=10.9Hz,1H),7.42(dd,J=13.6,7.7Hz,2H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),4.94(dd,J=12.2,5.3Hz,1H),4.47(br,1H),4.31(ddd,J=13.6,9.9,3.5Hz,1H),4.09–3.96(m,1H),3.87(d,J=10.5Hz,2H),3.61(d,J=10.4Hz,2H),3.36(br,4H),2.97–2.64(m,7H),2.53(d,J=4.5Hz,2H),2.23–2.10(m,5H),1.66(dd,J=12.7,2.7Hz,3H),1.54–1.37(m,3H).
177:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺
LC/MS(ESI+)calcdfor C41H42ClN9O6(M+H)+m/z,792.3;found 792.3.
1H NMR(400MHz,CDCl3)δ8.84(d,J=1.2Hz,1H),8.47(s,1H),7.70(d,J=8.6Hz,2H),7.56(d,J=8.7Hz,1H),7.41(d,J=8.2Hz,1H),7.29(d,J=2.1Hz,1H),7.07(dd,J=8.6,2.2Hz,1H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.95(dd,J=12.3,5.3Hz,1H),4.60(br,2H),4.30(ddd,J=13.2,9.7,3.1Hz,1H),4.09–3.97(m,1H),3.87(d,J=10.3Hz,2H),3.61(d,J=10.5Hz,2H),3.47(br,4H),2.94–2.73(m,3H),2.71(br,4H),2.47(d,J=6.5Hz,2H),2.24–2.08(m,5H),1.71(dd,J=13.6,3.7Hz,3H),1.54–1.39(m,2H).
178:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
1.化合物(1R,5S,6r)-3-氮杂双环[3.1.0]己烷-6-基甲醇的合成
将化合物(1R,5S,6r)-叔丁基6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(50mg,0.23mmol)溶于3mL二氯甲烷,并加入1.5mL三氟乙酸,室温搅拌反应2h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸,得化合物(1R,5S,6r)-3-氮杂双环[3.1.0]己烷-6-基甲醇三氟乙酸盐,不经进一步纯化,直接用于下步反应中。
2.化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
向装有(1R,5S,6r)-3-杂双环[3.1.0]己烷-6-基甲醇三氟乙酸盐的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(82mg,0.21mmol),碳酸钾(95mg,0.69mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。4h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(91mg,0.19mmol)。收率:83%。LC/MS(ESI+)calcd for C22H33N2O5(M+H)+m/z,467.9;found,468.1.
3.化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(91mg,0.19mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(161mg,0.38mmol),完毕,任体系自然升温至室温,并在室温搅拌反应过夜。翌日,TLC检测原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(76mg,0.16mmol)。收率:84%。
LC/MS(ESI+)calcd for C35H46ClN4O5(M+H)+m/z,465.9;found,466.1.
4.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺
称取N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(35mg,0.07mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL)和甲醇(1mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚啉-1,3-二酮(24mg,0.07mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(45mg,0.21mmol),任体系在室温搅拌反应过夜。翌日,取样点板,TLC显示反应结束。停止搅拌,旋蒸除去溶剂后,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(28mg,0.03mmol)。收率:45%。LC/MS(ESI+)calcd for C43H46ClN6O7(M+H)+m/z,810.3;found,810.3.
1H NMR(400MHz,CDCl3)δ8.44(s,1H),8.00(d,J=9.4Hz,1H),7.91(d,J=8.2Hz,1H),7.58(d,J=8.7Hz,1H),7.51(d,J=10.9Hz,1H),7.45(d,J=7.2Hz,1H),7.02(d,J=2.4Hz,1H),6.87(dd,J=8.7,2.4Hz,1H),6.71(d,J=9.4Hz,1H),4.96(dd,J=12.2,5.3Hz,1H),4.37–4.30(m,1H),4.15–4.00(m,1H),3.94(br,2H),3.68(d,J=9.4Hz,2H),3.41(s,4H),2.99–2.68(m,6H),2.60(br,2H),2.27–2.09(m,5H),1.78–1.65(m,6H),1.48(dd,J=22.4,10.3Hz,2H).
179:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C41H42ClN9O6(M+H)+m/z,792.3;found,792.3.
1H NMR(400MHz,CDCl3)δ8.35(s,1H),7.98(d,J=9.3Hz,1H),7.90(d,J=8.0Hz,1H),7.71(d,J=8.4Hz,1H),7.56(d,J=8.7Hz,1H),7.30(s,1H),7.08(d,J=8.4Hz,1H),7.00(d,J=2.0Hz,1H),6.86(dd,J=8.7,2.1Hz,1H),6.69(d,J=9.4Hz,1H),4.95(dd,J=11.9,5.0Hz,1H),4.40–4.24(m,1H),4.15–3.99(m,1H),3.91(br,2H),3.66(d,J=8.7Hz,2H),3.51(br,4H),2.82(ddd,J=26.2,22.5,13.7Hz,7H),2.52(br,2H),2.18(br,5H),1.69(dd,J=22.6,11.7Hz,5H),1.46(dd,J=22.4,10.6Hz,2H).
180:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基哒嗪-3-甲酰胺
向装有哌啶-4-基甲醇(82mg,0.71mmol)的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(250mg,0.64mmol),碳酸钾(294mg,2.13mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。4h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经柱层析分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基哒嗪-3-甲酰胺(218mg)。收率:65%。LC/MS(ESI+)calcd for C24H28ClN5O3(M+H)+m/z,470.0;found,469.9.
2.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基哒嗪-3-甲酰胺(50mg,0.11mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(72mg,0.17mmol),完毕,任体系自然升温至室温搅拌反应。2h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(30mg)。收率:58%。LC/MS(ESI+)calcd for C24H26ClN5O3(M+H)+m/z,468.0;found,467.9.
3.化合物叔丁基-6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯的合成
将2-(2,6-二氧代哌啶-3-基)-5,6-双氟异吲哚啉-1,3-二酮(200mg,0.68mmol),叔丁基-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(144mg,0.68mmol)和DIPEA(264mg,2.04mmol)加入到5mL DMSO。加热至140℃,搅拌1h。冷却至室温,加水和乙酸乙酯萃取,有机层用0.1N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,粗品经柱层析分离纯化得化合物叔丁基-6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(325mg)。收率:98%。LC/MS(ESI+)calcd for C24H27FN4O6(M+H)+m/z,486.5;found,486.9,430.9.
4.2-(2,6-二氧代哌啶-3-基)-5-氟-6-(2,6-双氮杂螺环[3.4]辛烷-6-基)异吲哚啉-1,3-二酮
将化合物叔丁基-6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(200mg,0.41mmol)溶于5mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应3h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物2-(2,6-二氧代哌啶-3-基)-5-氟-6-(2,6-双氮杂螺环[3.4]辛烷-6-基)异吲哚啉-1,3-二酮(142mg)。收率:90%。
LC/MS(ESI+)calcd for C19H19FN4O4(M+H)+m/z,386.4;found,387.0,.
5.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(30mg,0.06mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧代哌啶-3-基)-5-氟-6-(2,6-双氮杂螺环[3.4]辛烷-6-基)异吲哚啉-1,3-二酮(24mg,0.06mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(38mg,0.18mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(21mg)。收率:39%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.8.
1H NMR(400MHz,CDCl3)δ8.37(s,1H),7.97(d,J=9.5Hz,1H),7.88(d,J=8.0Hz,1H),7.56(d,J=8.7Hz,1H),7.40(d,J=12.2Hz,1H),7.10–6.91(m,3H),6.86(dd,J=8.7,2.2Hz,1H),4.92(dd,J=12.0,5.0Hz,1H),4.50(d,J=13.2Hz,2H),4.32(t,J=9.6Hz,1H),4.12–3.99(m,1H),3.73(s,2H),3.59(br,2H),3.35(br,4H),3.02(t,J=11.9Hz,2H),2.94–2.66(m,3H),2.49(br,2H),2.29–2.08(m,7H),1.91(d,J=11.9Hz,3H),1.68(dd,J=22.2,10.1Hz,3H),1.46(dd,J=22.2,10.3Hz,3H).
181:N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
1.化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯腈的合成
将化合物((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨基甲酸叔丁酯(2400mg,6.33mmol)溶于24mL二氯甲烷中,并加入6mL三氟乙酸,室温搅拌反应6h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入30mL二氯甲烷和15mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷/甲醇(10:1)洗涤(30mL*3),合并有机层,依次用水,饱和食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯腈(1400m g)。收率:79%。
2.化合物6-氯-N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)哒嗪-3-甲酰胺的合成
称取6-氯哒嗪-3-羧酸(143mg,0.90mmol)置于50mL单颈圆底烧瓶中,并加入10mL二氯甲烷,室温搅拌均匀。随后将体系移至于冰水浴中继续降温冷却搅拌,并加入HATU(513mg,1.35mmol)和DIPEA(233.0mg,1.80mmol)。完毕,体系保温搅拌20min,再向体系中加入4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯腈(250mg,0.90mmol)。体系在冰水浴中搅拌反应。4h后,TLC监测原料消耗完毕,停止反应。向体系中加入二氯甲烷(10mL)和水(10mL),剧烈搅拌,后静置分层,水相用二氯甲烷反萃(5mL*3),合并有机相,依次用水,饱和食盐水洗涤,无水硫酸钠干燥,旋除溶剂得粗品,后经柱层析分离得化合物6-氯-N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)哒嗪-3-甲酰胺(312mg)。收率:82%。LC/MS(ESI+)calcd for C20H20Cl2N4O2(M+H)+m/z,419.3;found,418.9.
3.化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
向装有((1R,5S,6r)-3-氮杂双环[3.1.0]己烷-6-基)甲醇三氟乙酸盐(80mg,0.35mmol)的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入6-氯-N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)哒嗪-3-甲酰胺(130mg,0.31mmol),碳酸钾(242mg,1.75mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。5h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经柱层析分离纯化得化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(118mg)。收率:77%。
LC/MS(ESI+)calcd for C26H30ClN5O3(M+H)+m/z,496.0;found,495.9.
4.化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
将N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(60mg,0.12mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(102mg,0.24mmol),完毕,任体系自然升温至室温搅拌反应。5h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(40mg)。收率:67%。
LC/MS(ESI+)calcd for C26H28ClN5O3(M+H)+m/z,494.0;found,493.9,.
5.化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺的合成
称取N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(40mg,0.08mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧代哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚啉-1,3-二酮(29mg,0.08mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(51mg,0.24mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(23mg)。收率:34%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ8.59(br,1H),8.18(d,J=9.1Hz,1H),7.99(d,J=9.4Hz,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=10.9Hz,1H),7.43(d,J=7.2Hz,1H),6.97(d,J=2.3Hz,1H),6.81(dd,J=8.7,2.3Hz,1H),6.71(d,J=9.4Hz,1H),4.94(dd,J=12.1,5.3Hz,1H),4.19(d,J=9.1Hz,1H),4.07(s,1H),3.92(br,2H),3.67(d,J=9.1Hz,2H),3.32(br,4H),2.93–2.68(m,7H),2.50(d,J=6.5Hz,2H),2.24–2.20(m,1H),2.19–2.09(m,2H),2.01(dd,J=12.9,7.4Hz,1H),1.28(s,6H),1.21(s,6H).
182:N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-((1R,5S,6S)-6-((4-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)吡嗪-2-甲酰胺
LC/MS(ESI+)calcdfor C43H45ClFN9O6(M+H)+m/z,838.3;found,837.8.
1H NMR(400MHz,CDCl3)δ8.83(s,1H),8.47(br,1H),7.77(d,J=8.9Hz,1H),7.72(s,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=11.0Hz,1H),7.43(d,J=7.2Hz,1H),6.97(d,J=2.3Hz,1H),6.81(dd,J=8.7,2.3Hz,1H),4.94(dd,J=12.2,5.3Hz,1H),4.14(d,J=8.8Hz,1H),4.07(s,1H),3.87(d,J=9.9Hz,2H),3.62(d,J=10.2Hz,2H),3.32(br,4H),2.95–2.64(m,7H),2.48(d,J=6.4Hz,2H),2.21(dd,J=17.9,9.8Hz,1H),2.19–2.09(m,2H),2.01(ddd,J=10.1,8.4,2.8Hz,1H),1.26(s,6H),1.21(s,6H).
183:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)哒嗪-3-甲酰胺的合成
1.2-(2,6-二氧代哌啶-3-基)-5-氟-6-(4-(羟甲基)哌啶-1-基)异吲哚啉-1,3-二酮的合成
将2-(2,6-二氧代哌啶-3-基)-5,6-双氟异吲哚啉-1,3-二酮(294mg,1.00mmol),哌啶-4-基甲醇(115mg,1.00mmol)和DIPEA(388mg,3.00mmol)加入到5mL DMSO。加热至140℃,搅拌反应1h。冷却至室温,加水和乙酸乙酯萃取,有机层用0.1N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,粗品经柱层析分离纯化得化合物2-(2,6-二氧代哌啶-3-基)-5-氟-6-(4-(羟甲基)哌啶-1-基)异吲哚啉-1,3-二酮(311mg)。收率:80%。
LC/MS(ESI+)calcd for C19H20FN3O5(M+H)+m/z,389.4;found,389.7
2.1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-甲醛的合成
将2-(2,6-二氧代哌啶-3-基)-5-氟-6-(4-(羟甲基)哌啶-1-基)异吲哚啉-1,3-二酮(100mg,0.26mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(220mg,0.52mmol),完毕,任体系自然升温至室温搅拌反应。3h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-甲醛(63mg)。收率:63%。LC/MS(ESI+)calcd for C19H18FN3O5 +(M+H+)m/z,387.4;found,387.7,.
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)哒嗪-3-甲酰胺的合成
称取1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-甲醛(30mg,0.08mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2,6-双氮杂螺环[3.4]辛烷-6-基)哒嗪-3-甲酰胺(37mg,0.08mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(51mg,0.24mmol),任体系在室温搅拌反应。3h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)哒嗪-3-甲酰胺(21mg)。收率:31%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ8.59(br,1H),8.00(d,J=9.3Hz,1H),7.91(d,J=8.1Hz,1H),7.56(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.38(d,J=7.2Hz,1H),7.01(d,J=2.1Hz,1H),6.86(dd,J=8.7,2.2Hz,1H),6.71(d,J=9.4Hz,1H),4.94(dd,J=12.1,5.2Hz,1H),4.37–4.27(m,1H),4.12–3.99(m,1H),3.74(br,2H),3.64(d,J=11.2Hz,4H),3.38(dd,J=25.3,6.9Hz,4H),2.95–2.67(m,5H),2.52(d,J=6.6Hz,2H),2.31–2.28(m,2H),2.13(dd,J=14.2,6.3Hz,3H),1.86(d,J=11.9Hz,3H),1.76–1.54(m,4H),1.44(dt,J=21.3,11.1Hz,4H).
184:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
1.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基哒嗪-3-甲酰胺(80mg,0.17mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(110mg,0.26mmol),完毕,任体系自然升温至室温搅拌反应。2h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(60mg)。收率:75%。
LC/MS(ESI+)calcd for C24H26ClN5O3(M+H)+m/z,467.9;found,468.0.
2.化合物6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯的合成
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(60mg,0.13mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL),室温搅拌溶解澄清。随后向体系中加入2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯(28mg,0.13mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(83mg,0.39mmol),任体系在室温搅拌反应。4h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯(70mg)。收率:82%。
LC/MS(ESI+)calcd for C35H46ClN7O4(M+H)+m/z,664.2;found,664.7.
3.化合物6-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺的合成
将6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯(70mg,0.11mmol)溶于6mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应2h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物6-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(53mg)。收率:89%。LC/MS(ESI+)calcd for C30H38ClN7O2(M+H)+m/z,564.1;found,564.3,.
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
将6-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(28mg,0.05mmol),2-(2,6-二氧代哌啶-3-基)-5,6-双氟异吲哚啉-1,3-二酮(15mg,0.05mmol)和DIPEA(20mg,0.15mmol)加入到3mLDMSO。加热至140℃,搅拌反应1h。冷却至室温,加水和乙酸乙酯萃取,有机层用0.1N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,粗品经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(24mg)。收率:56%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ8.84(br,1H),7.97(d,J=9.6Hz,1H),7.88(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.37(d,J=10.7Hz,1H),7.00(d,J=2.3Hz,1H),6.98(d,J=9.7Hz,1H),6.92(d,J=7.5Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),4.92(dd,J=12.3,5.2Hz,1H),4.51(d,J=11.1Hz,2H),4.32(ddd,J=13.4,9.9,3.5Hz,1H),4.25–3.94(m,5H),3.04(t,J=12.2Hz,2H),2.96–2.63(m,6H),2.44(br,2H),2.29–2.05(m,7H),2.03–1.92(m,2H),1.77–1.60(m,6H),1.52–1.40(m,2H).
185:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H46ClN9O6(M+H)+m/z,820.3;found,819.8.
1H NMR(400MHz,CDCl3)δ8.53(br,1H),7.97(d,J=9.6Hz,1H),7.88(d,J=8.2Hz,1H),7.65(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.00(d,J=2.3Hz,1H),6.98(d,J=9.7Hz,1H),6.86(dd,J=9.0,2.2Hz,2H),6.53(dd,J=8.3,1.6Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.52(d,J=11.4Hz,2H),4.32(ddd,J=8.4,7.9,3.1Hz,1H),4.14–3.85(m,5H),3.04(t,J=11.8Hz,2H),2.80(dddd,J=32.8,29.2,14.3,4.0Hz,6H),2.44(br,2H),2.27–2.09(m,7H),2.03–1.93(m,2H),1.73–1.63(m,6H),1.51–1.41(m,2H).
186:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺的合成
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺(300mg,0.64mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(10mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(407mg,0.96mmol),完毕,任体系自然升温至室温搅拌反应。3h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(205mg)。收率:68%。
LC/MS(ESI+)calcd for C24H26ClN5O3(M+H)+m/z,467.9;found,468.0.
2.化合物6-((1-(5-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)吡嗪-2-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯的合成
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(70mg,0.15mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯(32mg,0.15mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(95mg,0.45mmol),任体系在室温搅拌反应。6h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物6-((1-(5-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)吡嗪-2-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸叔丁酯(80mg)。收率:80%。
LC/MS(ESI+)calcd for C35H46ClN7O4(M+H)+m/z,664.2;found,664.7.
3.化合物5-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺的合成
将叔丁基-6-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(70mg,0.11mmol)溶于5mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应2h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物5-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(57mg)。收率:91%。
LC/MS(ESI+)calcd for C30H38ClN7O2(M+H)+m/z,564.1;found,564.3,.
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺的合成
将5-(4-((2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(30mg,0.05mmol),2-(2,6-二氧代哌啶-3-基)-5,6-双氟异吲哚啉-1,3-二酮(15mg,0.05mmol)和DIPEA(19mg,0.15mmol)加入到3mLDMSO。加热至140℃,搅拌反应1h。冷却至室温,加水和乙酸乙酯萃取,有机层用0.1N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,粗品经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺(22mg)。收率:52%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ9.05(br,1H),8.83(s,1H),7.96(s,1H),7.56(d,J=8.7Hz,1H),7.38(dd,J=9.2,6.6Hz,2H),6.99(d,J=2.2Hz,1H),6.94(d,J=7.5Hz,1H),6.85(dd,J=8.7,2.2Hz,1H),4.92(dd,J=12.3,5.2Hz,1H),4.46(d,J=12.7Hz,2H),4.36–4.26(m,1H),4.08(tdd,J=22.2,13.9,8.5Hz,5H),2.97(t,J=12.6Hz,2H),2.92–2.62(m,6H),2.49–2.30(m,2H),2.25–2.07(m,7H),1.83(br,5H),1.75–1.60(m,3H),1.46(dt,J=14.7,7.3Hz,2H).
187:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-6-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,820.3;found,819.8.
1H NMR(400MHz,CDCl3)δ8.83(s,1H),8.61(br,1H),7.96(s,1H),7.65(d,J=7.9Hz,1H),7.56(d,J=8.9Hz,1H),7.39(d,J=9.0Hz,1H),6.99(s,1H),6.85(d,J=6.6Hz,2H),6.53(d,J=8.5Hz,1H),4.89(dd,J=12.3,5.2Hz,1H),4.47(d,J=12.7Hz,2H),4.30(br,1H),4.08–3.91(m,5H),2.97(t,J=12.6Hz,2H),2.91–2.59(m,6H),2.48–2.29(m,2H),2.23–2.04(m,7H),1.85(br,5H),1.73–1.58(m,3H),1.48(dt,J=14.7,7.3Hz,2H).
188:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺
1.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺向装有哌啶-4-基甲醇(66mg,0.57mmol)的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(200mg,0.51mmol),碳酸钾(236mg,1.71mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。4h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经柱层析分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺(132mg)。收率:55%。
LC/MS(ESI+)calcd for C24H28ClN5O3(M+H)+m/z,470.0;found,469.9.
2.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺(60mg,0.13mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。再将体系置于冰水浴中降温冷却搅拌,待体系内温降至约0℃时,向体系中加入戴斯马丁氧化剂(85mg,0.20mmol),完毕,任体系自然升温至室温搅拌反应。3h后,TLC显示原料基本消耗完全,停止反应。使用硅藻土,对体系进行抽滤操作,滤饼用二氯甲烷少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(41mg)。收率:67%。
LC/MS(ESI+)calcd for C24H26ClN5O3(M+H)+m/z,467.9;found,468.0.
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺的合成
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(22mg,0.05mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(4mL),室温搅拌溶解澄清。随后向体系中加入2-(2,6-二氧代哌啶-3-基)-5-氟-6-(2,6-双氮杂螺环[3.4]辛烷-6-基)异吲哚啉-1,3-二酮(19mg,0.05mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(32mg,0.15mmol),任体系在室温搅拌反应。5h后,取样点板,TLC监测原料消耗完全。停止搅拌,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((6-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-双氮杂螺环[3.4]辛烷-2-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺(10mg)。收率:24%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,837.8.
1H NMR(400MHz,CDCl3)δ8.82(s,1H),8.09(br,1H),7.95(s,1H),7.56(d,J=8.8Hz,1H),7.39(t,J=10.3Hz,2H),7.00(dd,J=8.6,4.9Hz,2H),6.85(dd,J=8.7,2.2Hz,1H),5.38–5.32(m,1H),4.92(dd,J=12.2,5.1Hz,1H),4.45(d,J=13.7Hz,2H),4.35–4.26(m,1H),4.08–3.98(m,1H),3.72(br,2H),3.59(dd,J=6.1,4.2Hz,2H),3.43–3.20(m,4H),3.01–2.90(m,2H),2.90–2.64(m,3H),2.45(d,J=6.2Hz,2H),2.27–2.10(m,7H),2.06–1.96(m,2H),1.87(d,J=13.3Hz,3H),1.46(dd,J=21.9,12.2Hz,3H).
189:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺
1.化合物叔丁基-6-(5-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)吡嗪-2-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯的合成
向装有叔丁基-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(55mg,0.26mmol)的圆底烧瓶中,加入5mL DMF,室温搅拌溶解澄清。随后向体系中加入5-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-2-甲酰胺(100mg,0.26mmol),碳酸钾(101mg,0.78mmol),完毕,对体系进行抽真空通氩气操作,反复5次,确保体系中的惰性气体氛围。然后,将体系移置于80℃的油浴中,加热搅拌反应。3h后,TLC检测反应结束。停止加热,待体系冷却至室温,向体系中加入乙酸乙酯(10mL)和水(15mL),剧烈搅拌,后静置分层。水相用乙酸乙酯(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物叔丁基-6-(5-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)吡嗪-2-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(80mg)。收率:54%。
LC/MS(ESI+)calcd for C29H35ClN6O4(M+H)+m/z,567.1;found,566.9.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺的合成
将叔丁基-6-(5-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰)吡嗪-2-基)-2,6-双氮杂螺环[3.4]辛烷-2-羧酸酯(80mg,0.14mmol)溶于5mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应2h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入10mL二氯甲烷和3mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(3mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,粗品经柱层析分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺(44mg)。收率:64%。
LC/MS(ESI+)calcd for C24H27ClN6O2(M+H)+m/z,467.0;found,467.7,.
3.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺(44mg,0.09mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-甲醛(35mg,0.09mmol)以及一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(57mg,0.27mmol),任体系在室温搅拌反应。4h后,取样点板,TLC显示反应结束。停止搅拌,旋蒸除去溶剂后,向体系中加入二氯甲烷(15mL)和水(15mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,6-双氮杂螺环[3.4]辛烷-6-基)吡嗪-2-甲酰胺(30mg)。收率:39%。
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H)+m/z,838.3;found,838.3.
1H NMR(400MHz,CDCl3)δ8.61(br,1H),8.02(d,J=9.3Hz,1H),7.89(d,J=8.1Hz,1H),7.54(d,J=8.7Hz,1H),7.43(d,J=11.0Hz,1H),7.35(d,J=7.2Hz,1H),6.99(d,J=2.1Hz,1H),6.83(dd,J=8.7,2.2Hz,1H),6.68(d,J=9.4Hz,1H),4.91(dd,J=12.1,5.2Hz,1H),4.33–4.23(m,1H),4.09–3.92(m,1H),3.71(br,2H),3.61(d,J=11.2Hz,4H),3.34(dd,J=25.3,6.9Hz,4H),2.89–2.62(m,5H),2.48(d,J=6.6Hz,2H),2.29–2.26(m,2H),2.11(dd,J=14.2,6.3Hz,3H),1.83(d,J=11.9Hz,3H),1.72–1.51(m,4H),1.48-1.38(m,4H).
190:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-双氮杂螺环[3.5]壬烷-7-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcdfor C44H48ClN9O6(M+H)+m/z,834.4;found833.9.
1H NMR(400MHz,CDCl3)δ8.60(br,1H),7.98(d,J=9.5Hz,1H),7.88(d,J=8.0Hz,1H),7.67(d,J=8.3Hz,1H),7.56(d,J=8.6Hz,1H),7.14–6.89(m,4H),6.85(d,J=8.4Hz,1H),4.94(dd,J=11.3,5.2Hz,1H),4.31(br,1H),4.00(dd,J=42.4,8.2Hz,4H),3.71(br,5H),3.22(br,5H),3.07–2.63(m,7H),2.50(br,3H),1.73–1.63(m,4H),1.53–1.39(m,3H),1.29(dt,J=24.2,12.5Hz,5H).
191:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcd for C46H49ClFN7O6(M+H)+m/z,850.4;found,850.3.
1H NMR(400MHz,CDCl3)δ8.20(br,1H),7.65(d,J=8.9Hz,2H),7.56(d,J=8.7Hz,1H),7.36(d,J=10.9Hz,1H),6.99(d,J=2.3Hz,1H),6.89(d,J=9.0Hz,2H),6.84(dd,J=8.8,2.4Hz,1H),6.81(d,J=7.6Hz,1H),5.86(d,J=7.7Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.28(ddd,J=13.8,10.3,3.7Hz,1H),4.11–3.97(m,1H),3.89(s,4H),3.82(d,J=12.7Hz,2H),2.95–2.65(m,5H),2.38(br,4H),2.24–2.09(m,7H),1.68(dd,J=23.2,10.2Hz,5H),1.48–1.33(m,3H),1.28(dt,J=19.6,7.3Hz,5H).
192:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcd for C46H50ClN7O6(M+H)+m/z,832.4;found,832.3.
1H NMR(400MHz,CDCl3)δ8.17(br,1H),7.65(dd,J=8.0,5.5Hz,3H),7.56(d,J=8.7Hz,1H),6.99(d,J=1.6Hz,1H),6.95–6.82(m,2H),6.77(s,1H),6.51(d,J=8.2Hz,1H),5.87(d,J=7.4Hz,1H),4.93(dd,J=11.6,4.4Hz,1H),4.33–4.24(m,1H),4.04(d,J=8.0Hz,1H),3.83(d,J=12.6Hz,2H),3.74(s,4H),3.03–2.60(m,5H),2.40(br,4H),2.29–2.03(m,8H),1.68(dd,J=22.7,10.5Hz,5H),1.35(ddd,J=33.0,21.7,9.7Hz,7H).
193:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-双氮杂螺环[3.5]壬烷-2-基)-1,3,4-噻二唑-2-甲酰胺
LC/MS(ESI+)calcd for C41H44ClN9O6S(M+H)+m/z,826.4;found,826.2.
1H NMR(400MHz,CDCl3)δ8.06(br,1H),7.69(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.4Hz,1H),7.06(dd,J=8.4,1.7Hz,1H),7.00(t,J=5.2Hz,2H),6.84(dd,J=8.7,2.3Hz,1H),5.41–5.28(m,1H),4.94(dd,J=12.2,5.2Hz,1H),4.31(t,J=10.0Hz,1H),4.02(d,J=12.4Hz,3H),3.91(s,4H),2.99(t,J=12.4Hz,2H),2.94–2.57(m,7H),2.26–2.07(m,6H),2.01(dd,J=12.5,6.5Hz,6H),1.54–1.40(m,4H).
194:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(7-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-双氮杂螺环[3.5]壬烷-2-基)苯甲酰胺
LC/MS(ESI+)calcd for C46H50ClN7O6(M+H)+m/z,832.4;found,832.3.
1H NMR(400MHz,CDCl3)δ8.26(br,1H),7.67(d,J=8.6Hz,1H),7.63(d,J=8.7Hz,2H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.2Hz,1H),7.04(dd,J=8.7,2.3Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),6.39(d,J=8.7Hz,2H),5.82(d,J=7.8Hz,1H),4.94(dd,J=12.3,5.4Hz,1H),4.28(ddd,J=13.7,10.3,3.7Hz,1H),4.10–3.99(m,1H),3.95(d,J=12.8Hz,2H),3.65(s,4H),3.07–2.63(m,6H),2.42(br,4H),2.28–2.09(m,8H),1.89–1.80(m,6H),1.67(dd,J=23.3,10.3Hz,3H),1.39(dd,J=23.7,10.4Hz,2H).
195:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1.化合物2-(溴甲基)-4,5-双氟苯甲酸甲酯的合成
称取4,5-双氟-2-甲基苯甲酸甲酯(3500mg,18.80mmol)置于100mL单颈圆底烧瓶中,并向其中加入25mL四氯化碳,室温搅拌溶解澄清。随后,依次向体系中加入NBS(3680mg,20.68mmol),过氧化苯甲酰(136mg,0.56mmol),完毕,将体系移置于70℃的油浴中,加热搅拌反应。翌日,TLC检测原料基本消耗完毕。停止加热,待体系冷却至室温,旋蒸除去溶剂,后向体系中加入二氯甲烷(30mL)和水(15mL),剧烈搅拌,后静置分层。水相用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经柱层析分离纯化得化合物2-(溴甲基)-4,5-双氟苯甲酸甲酯(3180mg)。收率:64%。
2.化合物3-(5,6-双氟-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮的合成
称取2-(溴甲基)-4,5-双氟苯甲酸甲酯(1500mg,5.66mmol)置于100mL单颈圆底烧瓶中,并向其中加入25mL乙腈,室温搅拌溶解澄清。随后,向体系中缓慢地加入3-氨基哌啶-2,6-二酮盐酸盐(931mg,5.66mmol),后向体系中滴加5mL溶有三乙胺(1145mg,11.32mmol)的乙腈溶液,完毕,将体系移置于80℃的油浴中,加热搅拌反应。5h后,LCMS监测反应结束。停止加热,待体系冷却至室温,旋蒸除去溶剂,向体系中加入二氯甲烷(50mL)和水(25mL),剧烈搅拌,后静置分层。水相用二氯甲烷(20mL*3)反萃,合并有机相,依次用水(15mL*2),饱和食盐水(20mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经柱层析分离纯化得化合物3-(5,6-双氟-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(664mg)。收率:42%。
LC/MS(ESI+)calcd for C13H10F2N2O3(M+H)+m/z,280.2;found,281.1,.
3.化合物2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯的合成
将3-(5,6-双氟-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(200mg,0.71mmol),2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(160mg,0.71mmol)和DIPEA(275mg,2.13mmol)加入到5mL DMSO,加热至140℃反应过夜。翌日,TLC监测原料消耗完毕。停止加热,任体系冷却至室温,加水和乙酸乙酯萃取,有机层用0.1N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂,粗品经Pre-TLC分离纯化得化合物2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(102mg)。收率:29%。
LC/MS(ESI+)calcd for C25H31FN4O5(M+H)+m/z,486.5;found,487.1.
4.化合物3-(6-氟-1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮
将化合物2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(65mg,0.13mmol)溶于5mL二氯甲烷,并加入2mL三氟乙酸,室温搅拌反应3h。TLC监测原料消耗完毕,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。再次加入15mL二氯甲烷和5mL水,后将体系移至于冰水浴中降温冷却搅拌,随后向体系中滴加饱和碳酸氢钠溶液调节体系的pH至9左右,完毕,静置分层,水相用二氯甲烷洗涤(5mL*3),合并有机层,再用食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂,粗品经Pre-TLC分离纯化得化合物3-(6-氟-1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮(49mg)。收率:94%。
LC/MS(ESI+)calcd for C20H23FN4O3(M+H)+m/z,386.4;found,387.1.
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(61mg,0.13mmol)以及3-(6-氟-1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮(49mg,0.13mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(5mL),室温搅拌溶解澄清。随后向体系中加入一滴冰醋酸,完毕,体系在室温搅拌反应15min。尔后,向体系中加入三乙酰基硼氢化钠(83mg,0.39mmol),任体系在室温搅拌反应。6h后,取样点板,TLC显示反应结束。向体系中加入二氯甲烷(20mL)和水(10mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(35mg)。收率:33%。LC/MS(ESI+)calcd for C44H49ClFN9O5(M+H)+m/z,838.4;found,838.3.
1H NMR(400MHz,CDCl3/CD3OD)δ7.97(dd,J=8.6,4.8Hz,1H),7.57(d,J=8.7Hz,1H),7.40(d,J=11.3Hz,1H),7.02(dd,J=5.9,3.5Hz,2H),6.87(dd,J=8.8,2.4Hz,1H),6.41(d,J=7.6Hz,1H),5.13(dd,J=13.3,5.1Hz,1H),4.51(d,J=12.9Hz,2H),4.33(dd,J=15.8,9.5Hz,2H),4.22(d,J=15.8Hz,1H),4.04(ddd,J=10.4,7.9,4.0Hz,1H),3.84(br,4H),3.40(dt,J=3.2,1.6Hz,1H),3.07(t,J=12.3Hz,2H),2.96–2.74(m,3H),2.31(ddd,J=25.5,12.5,5.6Hz,3H),2.18(d,J=10.6Hz,6H),1.99(br,6H),1.69(dd,J=22.1,10.1Hz,3H),1.49(dd,J=22.2,10.9Hz,3H).
196:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
1.2-(1-氧代-1,3-二氢异苯并呋喃-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯
分别称取5-溴异苯并呋喃-1(3H)-酮(2500mg,11.74mmol),2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(2657mg,11.74mmol),Pd2(dba)3(540mg,0.59mmol),Xantphos(341mg,0.59mmol)和磷酸钾(4984mg,23.48mmol)置于150mL单颈圆底烧瓶中,并向其中加入1,4-二氧六环(40mL),室温搅拌均匀,对体系进行抽真空通氩气操作,反复7次,确保体系中的惰性气体氛围。然后,将体系移置于100℃的油浴中,加热搅拌反应过夜。翌日,TLC监测原料消失。停止加热,待体系冷却至室温,对体系进行抽滤操作(通过硅藻土),滤饼用二氧六环少量多次淋洗,合并滤液,旋蒸除去溶剂得粗品。经打浆(乙酸乙酯/石油醚=1/2)后得化合物2-(1-氧代-1,3-二氢异苯并呋喃-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(1400mg)。收率:33%。LC/MS(ESI+)calcd for C20H26N2O4(M+H)+m/z,358.4;found,359.2.
2.化合物4-(7-(叔丁氧羰基)-2,7-双氮杂螺环[3.5]壬烷-2-基)-2-(羟甲基)苯甲酸的合成
称取2-(1-氧代-1,3-二氢异苯并呋喃-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(1400mg,3.91mmol)置于100mL单颈圆底烧瓶中,并向其中加入18mL甲醇以及18mL四氢呋喃,室温搅拌均匀。随后向体系中滴加5mL溶有氢氧化钠(626mg,15.64mmol)的水溶液,完毕,将体系移置于45℃的油浴中,加热搅拌反应过夜。翌日,TLC监测反应结束。停止加热,待体系冷却至室温,旋蒸除去大部分溶剂,加入10mL水,随后将体系移置于冰水浴中降温冷却搅拌,10min后,向体系中滴加稀盐酸(0.5N)调节pH值至4-5左右,尔后向体系中加入乙酸乙酯,剧烈搅拌,静置分层,水相用乙酸乙酯反萃,合并有机层,再用食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品。经打浆(乙酸乙酯/石油醚=1/2)后得化合物4-(7-(叔丁氧羰基)-2,7-双氮杂螺环[3.5]壬烷-2-基)-2-(羟甲基)苯甲酸(874mg)。收率:59%。LC/MS(ESI+)calcd for C20H28N2O5(M+H)+m/z,376.4;found,377.2,321.1,277.1.
3.化合物2-(3-(羟甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯的合成
称取4-(7-(叔丁氧羰基)-2,7-双氮杂螺环[3.5]壬烷-2-基)-2-(羟甲基)苯甲酸(700mg,1.86mmol)置于50mL单颈圆底烧瓶中,并向其中加入8mL甲醇以及8mL乙酸乙酯,室温搅拌均匀。随后将体系移置于冰盐浴中降温冷却搅拌,待体系内温将至-10℃时,向体系中滴加3mL三甲基硅基重氮甲烷(2.0N),完毕,体系保温搅拌反应。30min后,TLC监测原料消耗完毕。向体系中加入10mL水和20mL乙酸乙酯,剧烈搅拌,静置分层,水相用乙酸乙酯反萃,合并有机层,再用食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品。经柱层析分离纯化得化合物2-(3-(羟甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(676mg)。收率:93%。LC/MS(ESI+)calcd for C21H30N2O5(M+H)+m/z,390.5;found,391.2,335.1,291.2.
4.2-(3-(溴甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯
称取2-(3-(羟甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(676mg,1.73mmol)置于50mL单颈圆底烧瓶中,并向其中加入10mL四氢呋喃,室温搅拌溶解澄清。随后向体系中依次加入三苯基膦(682mg,2.60mmol)和四溴化碳(862mg,2.60mmol),完毕,体系在室温搅拌反应。4h后,TLC监测反应结束。向体系中加入10mL水和30mL乙酸乙酯,剧烈搅拌,静置分层,水相用乙酸乙酯反萃,合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,旋蒸除去溶剂得粗品。经柱层析分离纯化得化合物2-(3-(溴甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(147mg)。收率:19%。
LC/MS(ESI+)calcd for C21H29BrN2O4(M+H)+m/z,453.4;found,353.1.
5.2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯
称取3-氨基哌啶-2,6-二酮盐酸盐(53mg,0.32mmol)置于25mL单颈圆底烧瓶中,并向其中加入4mL乙腈,室温搅拌均匀。接着向体系中缓慢滴加1mL溶有三乙胺(65mg,0.64mmol)的乙腈溶液,5min后,向体系中加入1mL溶有2-(3-(溴甲基)-4-(甲氧基羰基)苯基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(147mg,0.32mmol)的乙腈溶液,完毕,将体系移置于80℃的油浴中,加热搅拌反应。48h后,LCMS监测反应结束。停止加热,待体系冷却至室温,旋蒸除去溶剂,向体系中加入二氯甲烷(20mL)和水(10mL),剧烈搅拌,后静置分层。水相用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(35mg)。收率:23%。
LC/MS(ESI+)calcd for C25H32N4O5(M+H)+m/z,468.5;found,469.2,.
6.化合物3-(1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮的合成
将化合物2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-羧酸叔丁酯(35mg,0.07mmol)溶于3mL二氯甲烷,并加入1mL三氟乙酸,室温搅拌反应3h。TLC监测反应结束,旋蒸除去多余的三氟乙酸以及溶剂,并用二氯甲烷多次旋带除去残留的三氟乙酸。得化合物3-(6-氟-1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮三氟乙酸盐(39mg)。不经进一步纯化,直接用于下步反应中。
7.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
称取N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-甲酰胺(37mg,0.08mmol)以及3-(6-氟-1-氧代-5-(2,7-双氮杂螺环[3.5]壬烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮三氟乙酸盐(39mg,0.08mmol)置于25mL单颈圆底烧瓶中,并向其中加入二氯甲烷(3mL)和甲醇(1mL),体系在室温搅拌反应15min,随后,向体系中加入三乙酰基硼氢化钠(51mg,0.24mmol),任体系在室温搅拌反应。5h后,TLC监测原料消耗完毕。旋蒸除去溶剂,向体系中加入二氯甲烷(20mL)和水(10mL),剧烈搅拌,后静置分层。水层用二氯甲烷(10mL*3)反萃,合并有机相,依次用水(10mL*2),饱和食盐水(15mL)洗涤,无水硫酸钠干燥,旋蒸除去溶剂,得粗品,后经Pre-TLC分离纯化得化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)-2,7-双氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(12mg)。收率:18%。
LC/MS(ESI+)calcd for C44H50ClN9O5(M+H)+m/z,820.4;found,820.3,.
1H NMR(400MHz,CDCl3)δ8.03(s,1H),7.97(d,J=9.6Hz,1H),7.87(d,J=8.2Hz,1H),7.69(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),6.99(dd,J=10.5,6.0Hz,2H),6.86(dd,J=8.7,2.3Hz,1H),6.45(dd,J=8.6,1.3Hz,1H),6.37(s,1H),5.19(dd,J=13.2,5.1Hz,1H),4.52(d,J=12.4Hz,2H),4.38(d,J=15.7Hz,1H),4.35–4.27(m,1H),4.23(d,J=15.7Hz,1H),4.05(ddd,J=11.0,9.6,5.1Hz,1H),3.69(br,4H),3.05(t,J=12.3Hz,2H),2.86(dddd,J=17.0,12.6,7.1,3.9Hz,2H),2.32(ddd,J=25.5,12.7,5.0Hz,3H),2.24–2.10(m,6H),2.10–1.82(m,7H),1.68(dd,J=21.7,9.6Hz,6H),1.46(dd,J=22.2,11.0Hz,3H).
197:N-((1r,4r)-4-(4-氰基-3-(甲氧基-d3)苯氧基)环己基)-3-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-三嗪-6-羧酰胺
LC/MS(ESI+)calcd for C44H48D3N10O7(M+H+)m/z,834.4;found 835.
198:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物2-氯-4-((1r,3r)-3-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
将化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(527mg,1.89mmol)、碳酸钾(392mg,2.84mmol)和10mL DMF依次加入,氮气保护,搅拌均匀后滴入2-(溴甲基)-6-氯烟酸甲酯(500mg,1.89mmol)的DMF溶液,室温搅拌2h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到中间体。中间体加入DIEA(734mg,1.48mmol)和5mL 1,4二氧六环,100℃反应2d,TLC确定反应终点硅胶柱层析分离得到中间体2-氯-4-((1r,3r)-3-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(536mg,1.25mmol)。收率:66%。
LC/MS(ESI+)calcd for C22H21Cl2N3O2(M+H+)m/z,430.1;found,430.1.
2.化合物2-氯-4-((1r,3r)-3-(2-(4-(羟甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
将化合物2-氯-4-((1r,3r)-3-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(150mg,0.35mmol)、4-哌啶甲醇(120mg,1.05mmol)、DIEA(225mg,1.74mmol)和3mL DMF依次加入反应器,氮气保护,100℃搅拌反应过夜。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。pre-TLC分离得到类白色固体化合物3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丙哌啶-5-基)丁-3-炔-1-基)氧基)-N-(2-(2,6-二氧哌啶-3-基)-1-氧代异丙哌啶-4-基)丙酰胺(97mg,0.19mmol)。收率:55%。
LC/MS(ESI+)calcd for C28H33ClN4O3(M+H+)m/z,509.2;found,509.2.
3.2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物3-((4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异丙哌啶-5-基)丁-3-炔-1-基)氧基)-N-(2-(2,6-二氧哌啶-3-基)-1-氧代异丙哌啶-4-基)丙酰胺(60mg,0.12mmol)加入2mL DCM溶清,加入DMP(60mg,1.4mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚-1,3-二酮(41mg,1.2mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(14mg,0.24mmol)室温搅拌反应30min,随后加入氰基硼氢化钠(30mg,0.47mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(22mg,0.19mmol)。收率:22%。
LC/MS(ESI+)calcd for C45H48ClFN8O6(M+H+)m/z,851.3;found,851.2.
1H NMR(400MHz,CDCl3)δ8.05(s,1H),7.74(d,J=8.8Hz,1H),7.50(d,J=8.7Hz,1H),7.46–7.38(m,1H),7.34(s,1H),6.92(t,J=3.9Hz,1H),6.76(dd,J=8.7,2.4Hz,1H),6.61(d,J=8.9Hz,1H),4.87(dd,J=12.3,5.3Hz,1H),4.52–4.46(m,2H),4.41(d,J=12.6Hz,3H),4.18(s,1H),3.68(s,2H),3.23(s,3H),2.84(ddd,J=21.4,21.0,7.8Hz,4H),2.76–2.63(m,2H),2.56(s,3H),2.17(dd,J=21.7,14.1Hz,2H),2.09(dd,J=11.2,4.6Hz,1H),2.02–1.70(m,4H),1.42–1.33(m,6H),1.17(s,6H).
199:2-氯-4-((1r,3r)-3-(5-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异氧吲哚啉-5-基)哌嗪-1-基)丁-1-炔-1-基)-1-氧异氧吲哚啉-2-基)-2,4,4-四甲基环丁氧基)苯甲腈的合成
1.2-氯-4-((1r,3r)-3-(5-(3-羟基-1-炔-1-基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
化合物3-丁炔-2-醇(178mg,2.53mmol),4-((1r,3r)-3-(5-溴-1-氧代异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苄腈(400mg,0.84mmol),[1,1'-双(二苯基膦基)二茂铁]二氯化钯(62mg,0.084mmol),碘化亚铜(48mg,0.25mmol),三乙胺1mL和甲苯3mL依次加入反应器,氮气保护,110℃反应过夜。TLC确定反应结束,过滤,滤饼用二氯甲烷淋洗,旋干,硅胶柱层析纯化,得到咖啡色固体2-氯-4-((1r,3r)-3-(5-(3-羟基-1-炔-1-基)-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(300mg,0.65mmol),收率:77%。
LC/MS(ESI+)calcd for C27H27ClN2O3(M+H+)m/z,463.2;found,463.1.
2.化合物4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)丁-3-炔-2-基-4-甲基苯磺酸酯的合成
化合物2-氯-4-((1r,3r)-3-(5-(3-羟基-1-炔-1-基)-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(50mg,0.11mmol),DMAP(1mg,0.11mmol),三乙胺(33mg,0.32mmol)和2mL DCM依次加入反应瓶,氮气保护,0℃加入TsCl(25mg,0.13mmol),逐渐恢复至室温搅拌反应过夜。TLC确定反应结束,加入水,二氯甲烷萃取3次,无水硫酸钠干燥,过滤旋干,pre-TLC分离得到类白色固体4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)丁-3-炔-2-基-4-甲基苯磺酸酯(16mg,0.026mmol),收率:24%。
LC/MS(ESI+)calcd for C34H33ClN2O5S(M+H+)m/z,617.2;found,617.2.
3.化合物2-氯-4-((1r,3r)-3-(5-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异氧吲哚啉-5-基)哌嗪-1-基)丁-1-炔-1-基)-1-氧异氧吲哚啉-2-基)-2,4,4-四甲基环丁氧基)苯甲腈的合成
化合物4-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚-5-基)丁-3-炔-2-基-4-甲基苯磺酸酯(16mg,0.026mmol),2-(2,6-二氧哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚-1,3-二酮(9mg,0.026mmol),DIEA(10mg,0.077mmol)和1mL DMF依次加入反应器,氮气保护,50℃反应过夜。TLC确定反应结束,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。pre-TLC分离得到淡黄色固体2-氯-4-((1r,3r)-3-(5-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异氧吲哚啉-5-基)哌嗪-1-基)丁-1-炔-1-基)-1-氧异氧吲哚啉-2-基)-2,4,4-四甲基环丁氧基)苯甲腈(5mg,0.006mmol),收率:24%。LC/MS(ESI+)calcd for C44H42ClFN6O6(M+H+)m/z,805.3;found,805.2.
1H NMR(400MHz,CDCl3)δ8.11(d,J=8.8Hz,1H),7.72(d,J=7.8Hz,1H),7.53–7.44(m,3H),7.41(d,J=11.1Hz,1H),7.34(d,J=7.3Hz,1H),6.92(d,J=2.4Hz,1H),6.77(dd,J=8.7,2.4Hz,1H),4.87(dd,J=12.3,5.2Hz,1H),4.58(s,2H),4.34(s,1H),4.25(s,1H),3.76(d,J=7.1Hz,1H),3.29(s,4H),2.83(dd,J=13.4,9.9Hz,3H),2.78–2.65(m,4H),2.11–2.04(m,1H),1.43(d,J=7.0Hz,3H),1.37(d,J=7.5Hz,6H),1.19–1.17(m,6H).
HC-2954-01
200:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
1.2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
将化合物4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(2.37g,9.45mmol)、碳酸钾(1.96g,14.18mmol)和40mL DMF依次加入,氮气保护,搅拌均匀后滴入2-(溴甲基)-6-氯烟酸甲酯(2.5g,9.45mmol)的DMF溶液,室温搅拌2h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到中间体。中间体加入DIEA(4.73mg,36.61mmol)和20mL 1,4二氧六环,100℃反应2d,TLC确定反应终点硅胶柱层析分离得到中间体2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(1.85g,4.60mmol)。收率:49%。
LC/MS(ESI+)calcd for C20H17Cl2N3O2(M+H+)m/z,402.1;found,402.1.
1H NMR(400MHz,CDCl3)δ8.00(d,J=8.0Hz,1H),7.50(d,J=8.7Hz,1H),7.39(d,J=8.0Hz,1H),6.94(d,J=2.3Hz,1H),6.80(dd,J=8.7,2.3Hz,1H),4.32(s,2H),4.27(dd,J=9.6,4.0Hz,2H),2.23(s,2H),1.98(d,J=7.0Hz,2H),1.67(dd,J=17.0,8.4Hz,4H).
2.2-氯-4-((1r,4r)-4-(2-(4-(羟甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
将化合物2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(900mg,2.24mmol)、4-哌啶甲醇(773mg,6.71mmol)、DIEA(1.45mg,11.19mmol)和10mL NMP依次加入反应器,氮气保护,100℃搅拌反应过夜。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到类白色固体化合物2-氯-4-((1r,4r)-4-(2-(4-(羟甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(1.05mg,2.18mmol)。收率:98%。
LC/MS(ESI+)calcd for C26H29ClN4O3(M+H+)m/z,481.2;found,481.2.
3.化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
将化合物2-氯-4-((1r,4r)-4-(2-(4-(羟甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(60mg,0.12mmol)加入2mL DCM溶清,加入DMP(60mg,1.4mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(哌嗪-1-基)异吲哚-1,3-二酮(43mg,0.092mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(10mg,0.17mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(71mg,0.33mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(32mg,0.39mmol)。收率:46%。
LC/MS(ESI+)calcd for C43H44ClFN8O6(M+H+)m/z,822.3;found,822.3.
1H NMR(400MHz,CDCl3)δ8.23(s,1H),7.81(d,J=8.8Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=10.9Hz,1H),7.40(d,J=6.8Hz,1H),7.00(d,J=2.4Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.66(d,J=8.9Hz,1H),4.95(d,J=5.5Hz,1H),4.47(d,J=13.0Hz,2H),4.27(dd,J=12.6,8.7Hz,2H),4.19(s,2H),3.33(s,4H),2.89(ddd,J=21.7,20.6,8.4Hz,4H),2.78(dd,J=17.9,8.4Hz,2H),2.72–2.49(m,3H),2.20(d,J=22.8Hz,4H),2.16–2.11(m,1H),2.01(t,J=11.6Hz,5H),1.70(s,6H).
201:2-氯-4-((1r,4r)-4-(2-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)calcd for C43H45ClN8O6(M+H+)m/z,805.3;found,805.3.
1H NMR(400MHz,CDCl3)δ8.11(s,1H),7.74(d,J=8.8Hz,1H),7.64(d,J=8.4Hz,1H),7.49(d,J=8.7Hz,1H),7.23(s,1H),7.00(d,J=8.5Hz,1H),6.94(d,J=2.3Hz,1H),6.79(dd,J=8.8,2.4Hz,1H),6.59(d,J=8.9Hz,1H),4.88(dd,J=12.1,5.2Hz,1H),4.40(d,J=12.4Hz,2H),4.23(d,J=7.0Hz,2H),4.13(s,2H),3.38(s,3H),2.95–2.68(m,5H),2.53(s,3H),2.17(s,4H),2.11–2.03(m,1H),1.94(s,5H),1.62(d,J=7.9Hz,8H).
202:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
得到目标化合物为类白色固体化合物(17mg,0.20mmol)。收率:17%。
LC/MS(ESI+)calcd for C45H49ClN8O6(M+H+)m/z,833.4;found,833.3.
1H NMR(400MHz,CDCl3)δ8.11(s,1H),7.74(d,J=8.8Hz,1H),7.64(d,J=8.4Hz,1H),7.49(d,J=8.7Hz,1H),7.23(s,1H),7.00(d,J=8.5Hz,1H),6.94(d,J=2.3Hz,1H),6.79(dd,J=8.8,2.4Hz,1H),6.59(d,J=8.9Hz,1H),4.88(dd,J=12.1,5.2Hz,1H),4.40(d,J=12.4Hz,2H),4.23(d,J=7.0Hz,2H),4.13(s,2H),3.38(s,3H),2.95–2.68(m,5H),2.53(s,3H),2.17(s,4H),2.11–2.03(m,1H),1.94(s,5H),1.62(d,J=7.9Hz,8H).
203:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
204:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
将化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(57mg,0.070mmol)加入1mL HOAc溶清,加入Zn粉(92mg,1.41mmol)60℃搅拌反应过夜。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2mL DCM和三乙基硅烷(33mg,0.28mmol),随后加入1mL TFA室温搅拌反应过夜。TLC确定反应终点,减压蒸除溶剂,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC(EA:MeOH10:1)分离得到上点类白色固体化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈203(6mg,0.007mmol)。收率:11%。
LC/MS(ESI+)calcd for C43H46ClFN8O5(M+H+)m/z,810.3;found,810.3.
1H NMR(400MHz,CDCl3)δ9.16(s,0H),7.73(d,J=8.9Hz,1H),7.49(d,J=8.7Hz,1H),7.40(d,J=7.9Hz,1H),7.07(d,J=11.3Hz,1H),6.95(t,J=4.1Hz,1H),6.80(dd,J=8.7,2.4Hz,1H),6.60(d,J=8.9Hz,1H),5.09(dd,J=13.3,5.1Hz,1H),4.43–4.29(m,3H),4.21(d,J=16.1Hz,3H),4.13(s,2H),3.09(s,4H),2.88(dd,J=21.9,10.5Hz,2H),2.78(dt,J=18.1,5.2Hz,2H),2.59(s,4H),2.38–2.24(m,6H),2.21–2.08(m,4H),1.94(d,J=16.5Hz,2H),1.85(d,J=11.7Hz,3H),1.69–1.56(m,4H).
下点类白色化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈204(11mg,0.014mmol)。收率:19%。
LC/MS(ESI+)calcd for C43H46ClFN8O5(M+H+)m/z,810.3;found,810.3.
1H NMR(400MHz,CDCl3)δ9.57(s,0H),7.73(d,J=8.9Hz,1H),7.50(d,J=8.7Hz,1H),7.41(d,J=11.3Hz,1H),6.95(d,J=2.3Hz,1H),6.92(d,J=7.2Hz,1H),6.80(dd,J=8.8,2.4Hz,1H),6.61(d,J=8.9Hz,1H),5.06(dd,J=13.3,5.1Hz,1H),5.06(dd,J=13.3,5.1Hz,1H),4.43–4.30(m,3H),4.21(d,J=16.0Hz,3H),4.14(s,2H),3.15(s,4H),2.98(s,2H),2.90(t,J=12.2Hz,2H),2.77(dd,J=11.2,4.4Hz,2H),2.60(s,4H),2.33–2.22(m,3H),2.13(ddd,J=12.6,11.1,7.8Hz,3H),1.85(d,J=12.1Hz,3H),1.70–1.55(m,4H).
205:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
206:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
将化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(47mg,0.055mmol)加入1mL HOAc溶清,加入Zn(92mg,1.41mmol)60℃搅拌反应过夜。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2mL DCM和三乙基硅烷(33mg,0.28mmol),随后加入1mL TFA室温搅拌反应过夜。TLC确定反应终点,减压蒸除溶剂,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC(EA:MeOH 10:1)分离得到上点类白色固体化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(18mg,0.022mmol)。收率:39%。
LC/MS(ESI+)calcd for C45H50ClFN8O5(M+H+)m/z,837.4;found,837.4.
1H NMR(400MHz,CDCl3)δ7.81(d,J=8.8Hz,1H),7.58(d,J=8.7Hz,1H),7.48(d,J=7.9Hz,1H),7.15(d,J=11.2Hz,1H),6.99(d,J=2.2Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),6.69(d,J=8.9Hz,1H),5.17(dd,J=13.3,5.0Hz,1H),4.55(s,2H),4.45(d,J=12.0Hz,3H),4.35(d,J=36.4Hz,2H),4.25(s,1H),3.17(s,3H),3.05–2.92(m,3H),2.91–2.87(m,1H),2.68(s,4H),2.33(d,J=6.1Hz,3H),1.93(d,J=11.4Hz,3H),1.44(d,J=9.2Hz,6H),1.36(s,2H),1.29(s,2H),1.23(s,6H).
下点类白色固体化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(8mg,0.010mmol)。收率:17%。
LC/MS(ESI+)calcd for C45H50ClFN8O5(M+H+)m/z,837.4;found,837.4.
1H NMR(400MHz,CDCl3)δ7.81(d,J=8.8Hz,1H),7.58(d,J=8.7Hz,1H),7.48(d,J=7.9Hz,1H),7.14(s,1H),6.88(d,J=10.2Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),6.69(d,J=8.9Hz,1H),5.17(dd,J=13.3,5.0Hz,1H),4.55(s,2H),4.45(d,J=12.0Hz,3H),4.35(d,J=36.4Hz,2H),4.25(s,1H),3.17(s,3H),3.05–2.92(m,3H),2.91–2.87(m,1H),2.68(s,4H),2.33(d,J=6.1Hz,3H),1.93(d,J=11.4Hz,3H),1.44(d,J=9.2Hz,6H),1.36(s,2H),1.29(s,2H),1.23(s,6H).
207:(3-(5-(4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)哌啶-4-基)甲基)哌嗪-1-基)-6-氟-1-氧异靛-2-基)-2,6-二氧哌啶-1-基)甲基异丙基碳酸酯
1.化合物氯甲基异丙基碳酸二酯的合成
将化合物氯甲酸氯甲酯(2g,15.51mmol)加入100mL DCM溶清,0℃加入三乙胺(1.57g,15.51mmol)和异丙醇(932mg,15.51mmol)室温搅拌反应1h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。硅胶柱层析得到无色油状物氯甲基异丙基碳酸二酯(523mg,3.43mmol)。收率:22%。
LC/MS(ESI+)calcd for C5H9ClO3(M+H+)m/z,153.0;found,153.0.
2.(3-(5-(4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)哌啶-4-基)甲基)哌嗪-1-基)-6-氟-1-氧异靛-2-基)-2,6-二氧哌啶-1-基)甲基异丙基碳酸酯
将化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(10mg,0.012mmol)加入反应器,随后加入1mL DMF和碳酸钾(3mg,0.018mmol),氯甲基异丙基碳酸二酯(4mg,0.024mmol)用DMF溶清滴入反应液,室温搅拌反应2h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,有机层用饱和食盐水洗涤3次,无水硫酸钠干燥,旋干,pre-TLC分离得到类白色化合物(3-(5-(4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)哌啶-4-基)甲基)哌嗪-1-基)-6-氟-1-氧异靛-2-基)-2,6-二氧哌啶-1-基)甲基异丙基碳酸酯(2mg,0.002mmol)。收率:17%。
LC/MS(ESI+)calcd for C48H54ClFN8O8(M+H+)m/z,925.4;found,925.3.
208:2-氯-4-((1S,4r)-4-(2-((3S)-3-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)吡咯烷-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)calcd for C42H44ClFN8O5(M+H+)m/z,795.3;found,795.3.
1H NMR(400MHz,CDCl3)δ9.45(s,0H),7.81(d,J=8.7Hz,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=7.9Hz,1H),7.16(d,J=11.2Hz,1H),7.01(t,J=7.5Hz,1H),6.88(dd,J=8.8,2.4Hz,1H),6.42(d,J=8.8Hz,1H),5.16(dd,J=13.3,5.1Hz,1H),4.22(d,J=17.5Hz,2H),3.77(s,1H),3.67(d,J=16.9Hz,1H),3.49(t,J=16.0Hz,1H),3.35–3.24(m,1H),3.17(s,4H),2.90–2.81(m,2H),2.71(s,11H),2.52(dd,J=16.4,8.4Hz,2H),2.29–2.16(m,4H),2.00(s,2H),1.69(dd,J=20.3,11.0Hz,4H).
209:2-氯-4-((1S,4r)-4-(2-((3S)-3-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)吡咯烷-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
LC/MS(ESI+)calcd for C42H44ClFN8O5(M+H+)m/z,795.3;found,795.3.
1H NMR(400MHz,CDCl3)δ7.82(d,J=8.7Hz,1H),7.57(d,J=8.7Hz,1H),7.51(d,J=11.2Hz,1H),7.01(dd,J=8.6,4.8Hz,2H),6.87(dd,J=8.7,2.4Hz,1H),6.41(d,J=8.7Hz,1H),5.17(dd,J=13.3,5.1Hz,1H),4.42(d,J=16.0Hz,1H),4.34–4.26(m,3H),4.23(s,2H),3.79(s,1H),3.66(d,J=3.9Hz,1H),3.52(d,J=9.1Hz,1H),3.27(s,5H),2.90–2.49(m,9H),2.34(dd,J=13.0,5.5Hz,1H),2.30–2.14(m,5H),1.93–1.80(m,2H),1.70(s,4H).
210:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
211:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
212:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
1.2-氯-4-((1r,4r)-4-(5-氧-2-(哌嗪-1-基)-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈
将化合物2-氯-4-((1r,4r)-4-(2-氯-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(350mg,0.81mmol)、1-叔丁氧羰基哌嗪(454mg,2.44mmol)、DIEA(525mg,4.07mmol)和40mL NMP依次加入反应器,氮气保护,100℃搅拌反应过夜。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到中间体,中间体加入DCM/TFA溶清,室温搅拌反应1h。TLC确定反应终点,减压蒸除溶剂,加入饱和碳酸氢钠溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到类白色固体化合物2-氯-4-((1r,4r)-4-(5-氧-2-(哌嗪-1-基)-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(280mg,0.58mmol)。收率:72%。
LC/MS(ESI+)calcd for C24H26ClN5O2(M+H+)m/z,412.2;found,412.2.
2.2-氯-4-((1r,4r)-4-(2-(4-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌啶-4-基)甲基)哌嗪-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(210)
2-氯-4-((1r,4r)-4-(5-氧-2-(哌嗪-1-基)-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(100mg,0.26mmol)加入1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧哌啶-5-基)哌啶-4-乙醛(117mg,0.26mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(31mg,0.52mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(219mg,1.03mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(45mg,0.055mmol)。收率:21%。
LC/MS(ESI+)calcd for C43H44ClFN8O6(M+H+)m/z,823.3;found,823.2.
1H NMR(400MHz,CDCl3)δ8.28(s,1H),7.84(d,J=8.8Hz,1H),7.56(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.40(d,J=7.3Hz,1H),7.01(d,J=2.3Hz,1H),6.86(dd,J=8.7,2.3Hz,1H),6.66(d,J=8.8Hz,1H),4.94(dd,J=12.2,5.3Hz,1H),4.30(d,J=4.4Hz,2H),4.21(s,2H),3.74(s,3H),3.67(d,J=12.2Hz,2H),2.94–2.75(m,5H),2.61(s,4H),2.36(s,2H),2.24(s,2H),2.18–2.10(m,1H),2.05–1.94(m,4H),1.70(dd,J=16.5,8.1Hz,5H),1.54–1.36(m,3H).
3.2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(211)
将化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(57mg,0.070mmol)加入1mL AcOH溶清,加入Zn(92mg,1.41mmol)60℃搅拌反应过夜。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2mL DCM和三乙基硅烷(33mg,0.28mmol),随后加入1mL TFA室温搅拌反应过夜。TLC确定反应终点,减压蒸除溶剂,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC(EA:MeOH 20:1)分离得到上点类白色固体化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(17mg,0.021mmol)。收率:47%。
LC/MS(ESI+)calcd for C43H46ClFN8O5(M+H+)m/z,809.3;found,809.2.
1H NMR(400MHz,CDCl3)δ8.01(s,1H),7.86(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=7.7Hz,1H),7.12(t,J=9.9Hz,1H),7.00(t,J=4.4Hz,1H),6.86(dd,J=8.7,2.3Hz,1H),6.67(d,J=8.5Hz,1H),5.19(dd,J=13.2,5.1Hz,1H),4.34(dd,J=54.6,15.8Hz,4H),4.21(s,2H),3.73(s,3H),3.49(s,2H),2.99–2.83(m,2H),2.80–2.70(m,2H),2.59(s,3H),2.35(dt,J=13.0,8.2Hz,2H),2.30–2.19(m,3H),2.02(s,4H),1.77–1.57(m,7H).
212:下点类白色固体化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(5mg,0.006mmol)。收率:14%。LC/MS(ESI+)calcd for C43H46ClFN8O5(M+H+)m/z,809.3;found,809.2.
213:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)烟酰胺的合成
214:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)烟酰胺的合成
213:LC/MS(ESI+)calcd for C44H50ClFN8O5(M+H+)m/z,825.4;found,825.3.
1H NMR(400MHz,CDCl3)δ9.30(s,1H),8.55(d,J=2.2Hz,1H),7.90(dd,J=9.0,2.4Hz,1H),7.58(d,J=8.7Hz,1H),7.49(d,J=7.9Hz,1H),7.16(d,J=11.2Hz,1H),6.98(d,J=2.3Hz,1H),6.82(dd,J=8.7,2.3Hz,1H),6.69(d,J=9.1Hz,1H),6.23(d,J=8.3Hz,1H),5.16(dd,J=13.3,5.2Hz,1H),4.42(d,J=16.0Hz,3H),4.29(d,J=16.1Hz,1H),4.14(d,J=8.2Hz,1H),4.06(s,1H),3.19(s,3H),2.96(t,J=12.2Hz,3H),2.86(dd,J=12.7,5.2Hz,2H),2.71(s,4H),2.35(s,4H),2.22–2.15(m,1H),1.93(d,J=11.9Hz,3H),1.26(s,6H),1.22(s,6H).
214:LC/MS(ESI+)calcd for C44H50ClFN8O5(M+H+)m/z,825.4;found,825.3.
1H NMR(400MHz,CDCl3)δ8.55(d,J=2.2Hz,1H),7.90(dd,J=9.0,2.4Hz,1H),7.58(d,J=8.7Hz,1H),7.50(d,J=11.3Hz,1H),7.01–6.94(m,2H),6.81(dd,J=8.7,2.4Hz,1H),6.69(d,J=9.1Hz,1H),6.19(d,J=8.2Hz,1H),5.16(dd,J=13.3,5.1Hz,1H),4.47–4.37(m,3H),4.28(d,J=16.0Hz,1H),4.14(d,J=8.2Hz,1H),4.06(s,1H),3.21(s,4H),3.04–2.80(m,5H),2.66(s,4H),2.33(d,J=2.3Hz,4H),1.92(d,J=11.7Hz,4H),1.26(s,6H),1.21–1.20(m,6H).
215:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-(甲基-d3)烟酰胺的合成
216:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-(甲基-d3)烟酰胺的合成
215:LC/MS(ESI+)calcd for C45H49D3ClFN8O5(M+H+)m/z,842.4;found,842.3.
1H NMR(400MHz,CDCl3)δ8.28(s,1H),8.06(s,1H),7.57(d,J=8.6Hz,1H),7.49(d,J=8.7Hz,1H),7.41(d,J=7.8Hz,1H),7.07(d,J=11.1Hz,1H),6.90(d,J=2.3Hz,1H),6.73(dd,J=8.7,2.3Hz,1H),6.58(d,J=9.0Hz,1H),5.12(dd,J=13.3,5.1Hz,1H),4.35(d,J=15.8Hz,3H),4.21(d,J=15.9Hz,1H),4.06(s,1H),3.84(s,1H),3.12(s,4H),2.81(ddd,J=25.5,20.8,12.0Hz,4H),2.70–2.46(m,3H),2.28(ddd,J=25.8,12.7,4.9Hz,2H),2.15(dd,J=10.2,5.2Hz,1H),1.91(d,J=37.5Hz,3H),1.66(d,J=6.5Hz,3H),1.34(s,6H),1.19(s,6H).
216:LC/MS(ESI+)calcd for C45H49D3ClFN8O5(M+H+)m/z,842.4;found,842.3.
1H NMR(400MHz,CDCl3)δ8.28(s,1H),8.12(s,1H),7.57(d,J=7.4Hz,1H),7.49(d,J=8.7Hz,1H),7.43(d,J=11.3Hz,1H),6.90(d,J=6.9Hz,2H),6.73(dd,J=8.7,2.1Hz,1H),6.58(d,J=8.9Hz,1H),5.11(dd,J=13.2,5.0Hz,1H),4.34(d,J=15.3Hz,3H),4.19(d,J=15.9Hz,1H),4.06(s,1H),3.84(s,1H),3.18(s,4H),2.92–2.74(m,4H),2.63(s,3H),2.27(dd,J=13.0,4.9Hz,3H),2.19–2.09(m,1H),1.86(d,J=11.5Hz,3H),1.67(s,6H),1.34(s,6H),1.18(s,6H).
217:2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-4-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
1.化合物4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-4-基)哌嗪-1-羧酸叔丁酯的合成
将化合物2-(2,6-二氧代-哌啶-3-基)-4-氟-异吲哚-1,3-二酮(500mg,1.81mmol),1-叔丁氧羰基哌嗪(404mg,2.17mmol),DIEA(702mg,5.43mmol)和10mL DMSO依次加入反应器,氮气保护,130℃搅拌反应1h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到淡黄色固体化合物4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-4-基)哌嗪-1-羧酸叔丁酯(780mg,1.76mmol)。收率:97%。LC/MS(ESI+)calcd for C22H26N4O6(M+H+)m/z,443.2;found,443.2.
2.化合物2-(2,6-二氧哌啶-3-基)-4-(哌嗪-1-基)异吲哚-1,3-二酮的合成
将化合物4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-4-基)哌嗪-1-羧酸叔丁酯(110mg,0.25mmol)加入1mL DCM溶清,随后加入1mL TFA,室温搅拌反应1h。TLC确定反应终点,反应液蒸除溶剂,加入饱和碳酸氢钠水溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到淡黄色固体化合物2-(2,6-二氧哌啶-3-基)-4-(哌嗪-1-基)异吲哚-1,3-二酮(76mg,0.22mmol)。收率:89%。LC/MS(ESI+)calcd for C17H18N4O4(M+H+)m/z,343.1;found,343.1.
3.化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-4-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈的合成
将化合物2-氯-4-((1r,4r)-4-(2-(4-(羟甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(70mg,0.14mmol)加入2mL DCM溶清,加入DMP(74mg,0.17mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-4-(哌嗪-1-基)异吲哚-1,3-二酮(50mg,0.14mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(17mg,0.29mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(123mg,0.58mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-4-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(55mg,0.068mmol)。收率:47%。LC/MS(ESI+)calcd for C43H45ClN8O6(M+H+)m/z,805.3;found,805.3.
1H NMR(400MHz,CDCl3)δ7.74(dd,J=8.8,5.0Hz,1H),7.63–7.49(m,2H),7.37(s,1H),7.28–7.21(m,1H),7.17(d,J=5.4Hz,1H),7.02–6.93(m,1H),6.83(dd,J=7.4,4.8Hz,1H),6.64(dd,J=8.8,5.6Hz,1H),4.91(s,1H),4.73(s,1H),4.45–4.03(m,5H),3.67(s,6H),3.33(d,J=25.4Hz,4H),2.92(t,J=12.7Hz,2H),2.74(dd,J=38.7,12.5Hz,6H),2.31(s,1H),2.20(s,2H),2.05(s,1H),1.99–1.74(m,5H),1.64(s,2H),1.19(s,2H).
218:2-氯-4-((3aR,5r,6aR)-2-(6-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)辛基环戊烷[c]吡咯-5-基)氧基)苯甲腈
1.化合物(3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊二烯并[c]吡咯-2(1H)-羧酸叔丁酯
化合物叔丁基(3aR,5r,6aS)-5-羟基六氢环戊[c]吡咯-2(1H)-羧酸盐(500.0mg,2.20mmol),加入到2mL DMF中,冰水浴,加入NaH(106.0mg,2.6mmol),保温反应30分钟,将2-氯-4-氟苯腈(342.0mg,2.2mmol)用2mL DMF溶解,缓慢滴入,室温反应2小时。加入水和乙酸乙酯萃取,有机层再用饱和食盐水洗涤2次,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到白色固体化合物(3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊二烯并[c]吡咯-2(1H)-羧酸叔丁酯(550.0mg,1.52mmol),收率:69%。
LC/MS(ESI+)calcd for C19H22ClN2O3(M-56+H+)m/z,307.1;found,307.1.
2.化合物2-氯-4-((3aR,5r,6aS)-八氢环戊[C]吡咯-5-基)氧基)苯甲腈三氟乙酸盐的合成
化合物(3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊二烯并[c]吡咯-2(1H)-羧酸叔丁酯(520mg,1.40mmol)溶于2mL DCM和2mL TFA,室温搅拌反应1h。TLC确定反应终点,减压蒸除溶剂。得到化合物2-氯-4-((3aR,5r,6aS)-八氢环戊[C]吡咯-5-基)氧基)苯甲腈三氟乙酸盐(538mg,1.40mmol)粗品,直接投下一步反应。收率:100%。
LC/MS(ESI+)calcd for C14H14ClN2O(M+H+)m/z,263.1;found,263.1.
3.化合物2-氯-4-((((3Ar,5R,6aS)-2-(6-氯哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈
将化合物6-氯哒嗪-3-羧酸(159mg,1.0mmol)和HATU(400mg,1.10mmol)加入到5mLDCM,冰水浴,加入DIEA(388.8mg,3.10mmol),之后加入2-氯-4-((3aR,5r,6aS)-八氢环戊[C]吡咯-5-基)氧基)苯甲腈三氟乙酸盐(300.0mg,1.0mmol)。室温搅拌2h。加DCM和水萃取,无水硫酸钠干燥,旋干,硅胶柱层析纯化得到化合物淡黄色油状化合物2-氯-4-((((3Ar,5R,6aS)-2-(6-氯哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈(226mg,0.60mmol)。收率:56%。
LC/MS(ESI+)calcd for C19H16Cl2N4O2 +(M+H+)m/z,402.1;found,402.1.
4.化合物4-((1-(6-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊[c]吡咯-2-羰基)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯的合成
将化合物2-氯-4-((((3Ar,5R,6aS)-2-(6-氯哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈(220mg,0.50mmol),4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(186mg,0.60mmol)依次加入,加入3mL NMP,DIEA(211.5mg,1.6mmol)氮气保护,110℃搅拌反应过夜。加入水和乙酸乙酯萃取,有机层再用饱和食盐水洗涤,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化得到淡黄色油状化合物4-((1-(6-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊[c]吡咯-2-羰基)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(207.0mg,0.3mmol)。收率:58.3%。
LC/MS(ESI+)calcd for C34H44ClN7O4(M-56+H+)m/z,594.3;found,594.3.
4.化合物2-氯-4-((3aR,5r,6aS)-2-(6-(4-(哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈的合成
将化合物4-((1-(6-((3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)六氢环戊[c]吡咯-2-羰基)哒嗪-3-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(100.0mg,0.15mmol)溶于2mLDCM和2mL TFA,室温搅拌2h。溶剂旋干,得到化合物2-氯-4-((3aR,5r,6aS)-2-(6-(4-(哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈(102.0mg,0.15mmol)。收率:100.0%。
LC/MS(ESI+)calcd for C29H35ClN7O2(M+H+)m/z,550.3;found,550.3.
5.化合物2-氯-4-((3aR,5r,6aR)-2-(6-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)辛基环戊烷[c]吡咯-5-基)氧基)苯甲腈的合成
将2-氯-4-((3aR,5r,6aS)-2-(6-(4-(哌嗪-1-基甲基)哌啶-1-基)哒嗪-3-羰基)六氢环戊[c]吡咯-5-基)氧基)苯甲腈(102mg,0.15mmol),2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(47mg,0.17mmol)和DIEA(60mg,0.46mmol)加入到3mL DMSO。加热至130℃,搅拌过夜。冷却至室温,加水和乙酸乙酯萃取,有机层用0.5N HCl洗涤,再用食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物2-氯-4-((3aR,5r,6aR)-2-(6-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羰基)辛基环戊烷[c]吡咯-5-基)氧基)苯甲腈(56mg,0.07mmol)。收率:69.4%。
LC/MS(ESI+)calcd for C42H44ClN9O6 +(M+H+)m/z,806.3;found,806.2.
1H NMR(400MHz,CDCl3)δ8.26(s,1H),7.87(d,J=9.5Hz,1H),7.70(d,J=8.4Hz,1H),7.52(d,J=8.7Hz,1H),7.28(d,J=5.1Hz,1H),7.06(d,J=8.2Hz,1H),6.96(d,J=9.6Hz,1H),6.89(s,1H),6.77(d,J=8.2Hz,1H),4.95(dd,J=11.8,4.9Hz,1H),4.86(s,1H),4.52(d,J=12.2Hz,2H),4.28(dd,J=12.2,8.0Hz,1H),4.15(dd,J=12.3,3.6Hz,1H),3.91(dt,J=12.8,10.8Hz,2H),3.45(s,4H),3.03(t,J=12.2Hz,2H),2.95–2.70(m,5H),2.61(s,3H),2.43–2.21(m,4H),2.19–2.09(m,1H),1.91(dd,J=32.7,13.2Hz,5H),1.73(s,3H).
219:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcd for C43H45ClFN7O6([M+H]+)m/z:810.3;found 810.2.
220:N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-2-((3aR,6aS)-5-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)嘧啶-5-羧酰胺的合成
将化合物叔丁基(3aR,6aS)-5-(5-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)嘧啶-2-基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸盐(100mg,0.18mmol)加入1mL DCM溶清,随后加入1mL TFA,室温搅拌反应1h。TLC确定反应终点,反应液蒸除溶剂,加入饱和碳酸氢钠水溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到中间体。将中间体,2-(2,6-二氧哌啶-3-基)-5-氟-6-(4-氧哌啶-1-基)异吲哚-1,3-二酮(66mg,0.18mmol)加入2mL DCM/MeOH(1:1)溶清,加入冰醋酸(21mg,0.35mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(149mg,0.70mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到黄色固体化合物N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-2-((3aR,6aS)-5-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌啶-4-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)嘧啶-5-羧酰胺(19mg,0.023mmol)。收率:13%。
LC/MS(ESI+)calcd for C42H43ClFN9O6(M+H+)m/z,824.3;found,824.2.
1H NMR(400MHz,CDCl3)δ8.71(s,1H),7.57(d,J=8.7Hz,1H),7.46(d,J=10.9Hz,1H),7.38(d,J=7.3Hz,1H),7.01(d,J=2.3Hz,1H),6.87(dd,J=8.8,2.3Hz,1H),4.93(dd,J=12.0,5.4Hz,1H),4.30(t,J=10.5Hz,1H),3.99(t,J=11.1Hz,1H),3.83(s,1H),3.66(d,J=5.2Hz,2H),3.10(s,2H),2.89(dd,J=19.6,8.7Hz,2H),2.82–2.72(m,1H),2.67(s,1H),2.14(dd,J=13.6,6.2Hz,3H),2.04(s,1H),1.82(s,1H),1.66(dd,J=22.9,10.1Hz,1H),1.46(dd,J=20.5,9.6Hz,1H),1.29(s,1H).
221:N-((1r,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((3aR,6aS)-5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)哒嗪-3-羧酰胺的合成
将化合物N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-氧哌啶-1-基)哒嗪-3-甲酰胺(30mg,0.062mmol)和2-(2,6-二氧哌啶-3-基)-5-氟-6-((3aR,6aS)-六氢吡咯[3,4-c]吡咯-2(1H)-基)异吲哚啉-1,3-二酮(24mg,0.062mmol)依次加入反应器,随后加入2mL DCM/MeOH(1:1)溶清,加入冰醋酸(8mg,0.12mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(53mg,0.25mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到黄色固体化合物N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((3aR,6aS)-5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)哌啶-1-基)哒嗪-3-羧酰胺(44mg,0.052mmol)。收率:83%。
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found,852.3.
1H NMR(400MHz,CDCl3)δ8.30(s,1H),8.15(d,J=9.1Hz,1H),7.98(d,J=9.6Hz,1H),7.55(t,J=10.0Hz,1H),7.40(t,J=13.2Hz,1H),7.13(d,J=7.3Hz,1H),7.00(d,J=9.6Hz,1H),6.97(d,J=2.4Hz,1H),6.81(dd,J=8.7,2.4Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.41(d,J=13.1Hz,2H),4.18(d,J=9.0Hz,1H),4.07(s,1H),3.65(d,J=9.8Hz,2H),3.45(d,J=10.3Hz,2H),3.20(dd,J=12.6,9.9Hz,2H),3.03(s,4H),2.93–2.67(m,4H),2.61(d,J=4.7Hz,2H),2.52(s,1H),1.68(d,J=10.4Hz,2H),1.31–1.23(m,8H),1.21(s,6H).
222:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺的合成
1.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(羟甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺的合成
将化合物(3aR,5r,6aS)-5-(羟甲基)六氢环戊[c]吡咯-2(1H)-羧酸叔丁酯(161mg,0.67mmol)加入2mL DCM溶清,随后加入2mL TFA,室温搅拌反应1h。TLC确定反应终点,反应液减压蒸除溶剂,加入饱和碳酸氢钠水溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到粗品。粗品加入6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(218mg,0.56mmol),3mL DMF和DIPEA(216mg,1.67mmol),氮气保护,100℃搅拌反应过夜。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,无水硫酸钠干燥,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到类白色化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(羟甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(151mg,0.30mmol)。收率:55%。LC/MS(ESI+)calcd for C26H30ClN5O3(M+H+)m/z,496.2;found,496.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺的合成
将化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(羟甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(60mg,0.12mmol)加入2mL DCM溶清,加入DMP(61mg,1.4mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(六氢吡咯[3,4-c]吡咯-2(1H)-基)异吲哚-1,3-二酮(47mg,0.12mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(14mg,0.24mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(102mg,0.48mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(41mg,0.047mmol)。收率:39%。
LC/MS(ESI+)calcd for C45H47ClFN9O6(M+H+)m/z,864.3;found,864.1.
1H NMR(400MHz,CDCl3)δ8.32(s,0H),7.90(dd,J=9.4,2.5Hz,1H),7.84(d,J=8.2Hz,0H),7.49(d,J=8.7Hz,1H),7.36(dd,J=12.0,1.9Hz,1H),7.07(dd,J=7.3,3.3Hz,1H),6.93(d,J=2.3Hz,0H),6.79(dd,J=8.7,2.4Hz,0H),6.63(dd,J=9.4,4.0Hz,0H),4.85(dd,J=12.2,5.4Hz,0H),4.25(t,J=9.8Hz,0H),3.98(d,J=8.1Hz,0H),3.69(d,J=34.5Hz,1H),3.48(s,1H),3.41(t,J=10.4Hz,2H),3.00(s,2H),2.85–2.64(m,2H),2.47(s,1H),2.30–2.14(m,3H),1.75(d,J=6.0Hz,0H),1.63–1.57(m,1H),1.39(dd,J=21.6,9.8Hz,1H).
223:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(3-(5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)氮杂苷-1-基)哒嗪-3-羧酰胺的合成
将化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(3-氧杂氮杂环丁烷-1-基)哒嗪-3-甲酰胺(100mg,0.23mmol)和2-(2,6-二氧哌啶-3-基)-5-氟-6-((3aR,6aS)-六氢吡咯[3,4-c]吡咯-2(1H)-基)异吲哚啉-1,3-二酮(91mg,0.23mmol)依次加入反应器,随后加入2mL DCM/MeOH(1:1)溶清,加入冰醋酸(28mg,0.47mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(199mg,0.94mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(3-(5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)氮杂苷-1-基)哒嗪-3-羧酰胺(72mg,0.090mmol)。收率:38%。LC/MS(ESI+)calcd for C40H39ClFN9O6(M+H+)m/z,796.3;found796.2.
1H NMR(400MHz,CDCl3)δ8.54(s,1H),7.90(d,J=9.2Hz,1H),7.83(d,J=8.1Hz,1H),7.49(d,J=8.7Hz,1H),7.34(d,J=12.1Hz,1H),7.05(d,J=7.3Hz,1H),6.93(d,J=2.1Hz,1H),6.78(dd,J=8.7,2.1Hz,1H),6.53(d,J=9.2Hz,1H),4.83(dd,J=12.0,5.2Hz,1H),4.31–4.14(m,3H),4.12–4.01(m,2H),3.97(d,J=8.1Hz,1H),3.69–3.47(m,3H),3.38(d,J=10.1Hz,2H),2.97(s,2H),2.80(d,J=15.4Hz,3H),2.74–2.60(m,2H),2.51(d,J=7.1Hz,2H),2.21–1.99(m,5H),1.68–1.53(m,2H),1.39(dd,J=22.0,10.3Hz,2H).
224:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H+)m/z,838.3;found 837.7.
1H NMR(400MHz,CDCl3)δ8.73(s,1H),7.98(d,J=9.4Hz,1H),7.93(d,J=8.2Hz,1H),7.57(d,J=8.7Hz,1H),7.52–7.44(m,1H),7.41(dd,J=7.2,2.5Hz,1H),7.01(d,J=2.3Hz,1H),6.86(dd,J=8.7,2.3Hz,1H),6.72(d,J=9.4Hz,1H),4.95(dd,J=11.8,5.3Hz,1H),4.33(t,J=9.9Hz,1H),4.13–4.00(m,1H),3.83(s,1H),3.75(s,1H),3.52(d,J=9.0Hz,1H),3.43(d,J=10.0Hz,1H),3.31(s,4H),2.98(s,1H),2.94–2.84(m,2H),2.83–2.58(m,6H),2.48(d,J=26.8Hz,3H),2.17(dd,J=16.7,8.9Hz,6H),1.84(d,J=10.5Hz,2H),1.74–1.64(m,3H),1.47(dd,J=22.6,10.4Hz,2H).
225:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(3-(5-(2-(2,6-二氧哌啶-3-基)-6-
氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)氮杂苷-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C40H39ClFN9O6(M+H+)m/z,796.3;found,796.7.
1H NMR(400MHz,CDCl3)δ8.83(s,0H),8.42(s,0H),7.67–7.52(m,1H),7.42(t,J=9.2Hz,1H),7.14(d,J=7.3Hz,0H),7.00(d,J=2.2Hz,0H),6.86(dd,J=8.7,2.2Hz,0H),4.92(dd,J=12.1,5.2Hz,0H),4.37–4.22(m,1H),4.14(d,J=4.3Hz,1H),4.03(d,J=8.0Hz,0H),3.77–3.58(m,1H),3.48(d,J=10.1Hz,1H),3.07(s,1H),2.90(d,J=12.6Hz,1H),2.77(dd,J=17.1,7.5Hz,1H),2.60(d,J=7.4Hz,1H),2.17(dd,J=16.2,11.2Hz,2H),1.72–1.63(m,1H),1.47(dd,J=22.6,10.5Hz,1H).
226:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺的合成
1.化合物(3aR,5r,6aS)-5-((4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌嗪-1-基)甲基)六氢环戊[c]吡咯-2(1H)-羧酸叔丁酯的合成
将化合物2-氯-4-((1r,4r)-4-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)环己基)氧基)苯甲腈(130mg,0.29mmol),N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(哌嗪-1-基)哒嗪-3-甲酰胺,5-甲酰六氢环戊[c]吡咯-2(1H)2-羧酸叔丁酯(78mg,0.32mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(35mg,0.59mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(250mg,1.18mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,prep-TLC分离得到类白色固体化合物(3aR,5r,6aS)-5-((4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌嗪-1-基)甲基)六氢环戊[c]吡咯-2(1H)-羧酸叔丁酯(183mg,0.28mmol)。收率:93%。LC/MS(ESI+)calcd for C35H46ClN7O4(M+H+)m/z,664.3;found,664.3.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺的合成
将化合物(3aR,5r,6aS)-5-((4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌嗪-1-基)甲基)六氢环戊[c]吡咯-2(1H)-羧酸叔丁酯(80mg,0.12mmol)加入1mL DCM溶清,随后加入1mL TFA,室温搅拌反应1h。TLC确定反应终点,反应液蒸除溶剂,加入饱和碳酸氢钠水溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到类白色固体中间体。将中间体,2-(2,6-二氧代-哌啶-3-基)-5,6-二氟异吲哚啉-1,3-二酮(42mg,0.14mmol),DIEA(78mg,0.60mmol)和1mL DMSO依次加入反应器,氮气保护,130℃搅拌反应1h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。prep-TLC分离得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)甲基)哌嗪-1-基)哒嗪-3-甲酰胺(780mg,1.76mmol)。收率:97%。LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H+)m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ8.22(s,0H),8.01(d,J=9.3Hz,0H),7.89(d,J=8.2Hz,0H),7.56(d,J=8.7Hz,0H),7.41(d,J=12.2Hz,0H),7.08(d,J=7.3Hz,0H),6.98(dd,J=15.1,2.5Hz,1H),6.85(dd,J=8.7,2.1Hz,0H),4.92(dd,J=12.2,5.3Hz,0H),4.32(t,J=10.0Hz,0H),4.06(d,J=8.3Hz,0H),3.81(s,1H),3.72(s,0H),3.51(dd,J=22.5,7.9Hz,1H),3.34(d,J=10.7Hz,0H),2.90(d,J=13.6Hz,1H),2.85–2.72(m,1H),2.62(s,1H),2.43(s,1H),2.26–2.10(m,2H),1.79(s,0H),1.68(d,J=11.2Hz,2H),1.47(dd,J=22.3,10.4Hz,1H).
227:N-(4-(3-氯-4-氰基苯氧基)苯基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
LC/MS(ESI+)calcd for C41H37ClFN9O6(M+H+)m/z,806.2;found,805.7.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.87(s,1H),8.08–7.98(m,2H),7.97–7.88(m,2H),7.73(d,J=11.4Hz,1H),7.46(d,J=7.4Hz,1H),7.40(d,J=9.8Hz,1H),7.29(d,J=2.4Hz,1H),7.20(d,J=9.0Hz,2H),7.04(dd,J=8.7,2.4Hz,1H),5.76(s,1H),5.11(dd,J=12.8,5.4Hz,1H),4.54(d,J=12.1Hz,2H),3.26(s,4H),3.07(t,J=11.9Hz,2H),2.95–2.82(m,1H),2.56(dd,J=25.7,14.6Hz,5H),2.24(d,J=6.6Hz,2H),2.08–2.00(m,1H),1.94(s,1H),1.86(d,J=12.7Hz,2H),1.16(dd,J=21.3,10.8Hz,2H).
228:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-2-基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H+)m/z,838.3;found,837.7.
1H NMR(400MHz,CDCl3)δ8.98(s,1H),8.76(s,1H),7.89(s,1H),7.49(d,J=8.7Hz,1H),7.40(d,J=10.9Hz,1H),7.35–7.29(m,2H),6.92(d,J=2.3Hz,1H),6.78(dd,J=8.7,2.3Hz,1H),4.85(dd,J=12.2,5.4Hz,1H),4.32–4.13(m,2H),4.02–3.89(m,1H),3.09(dd,J=18.1,7.0Hz,8H),2.86–2.77(m,1H),2.69(td,J=15.4,4.1Hz,2H),2.43(s,1H),2.17–2.01(m,4H),1.89(s,3H),1.79(d,J=10.5Hz,2H),1.62(dd,J=22.4,9.8Hz,3H),1.46–1.32(m,4H).
229:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-2-基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C43H45ClN9O6(M+H+)m/z,820.3;found,819.8.
1H NMR(400MHz,CDCl3)δ8.84(d,J=1.2Hz,1H),8.61(s,1H),7.98(d,J=1.1Hz,1H),7.69(d,J=8.5Hz,1H),7.57(d,J=8.7Hz,1H),7.40(d,J=8.2Hz,1H),7.30(d,J=2.2Hz,1H),7.07(dd,J=8.6,2.3Hz,1H),7.01(d,J=2.4Hz,1H),6.86(dd,J=8.7,2.4Hz,1H),4.95(dd,J=12.3,5.3Hz,1H),4.40–4.23(m,3H),4.12–3.99(m,1H),3.49–3.32(m,4H),3.18(d,J=14.9Hz,6H),2.96–2.70(m,3H),2.50(s,1H),2.27–2.10(m,5H),1.89(d,J=14.4Hz,4H),1.84(s,2H),1.77–1.63(m,3H),1.48(dd,J=22.6,10.2Hz,4H).
230:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
1.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-甲酰胺
化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(1.00g,2.56mmol),4-羟甲基哌啶(883mg,7.67mmol),碳酸钾(1.77g,12.78mmol)和DMF12mL依次加入反应器,氮气保护,80℃反应3小时,TLC确定反应终点。确认反应完毕后,反应液加入水和乙酸乙酯萃取3次,合并乙酸乙酯层,并用饱和食盐水萃洗3次,干燥旋干,柱层析分离得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-甲酰胺(1.09g,2.32mmol),收率:91%。
LC/MS(ESI+)calcd for C24H27ClN5O3(M+H+)m/z,470.2;found,470.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-羧酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)哒嗪-3-甲酰胺(95mg,0.20mmol)溶于2mL DCM溶清后加入Dess-Martin试剂(128mg,0.30mmol),室温搅拌反应1小时,TLC确认反应终点。反应完毕,过滤滤饼用二氯甲烷淋洗,滤液用亚硫酸钠溶液萃洗,干燥旋干得到淡黄色固体,粗品直接投下一步反应。
LC/MS(ESI+)calcd for C24H25ClN5O3(M+H+)m/z,467.2;found,467.9.
3.化合物7-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬烷-2-羧酸叔丁酯的合成
将化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰基哌啶-1-基)哒嗪-3-羧酰胺粗品,加入1,2-二氯乙烷2mL溶清,并依次加入2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(55mg,0.24mmol),冰醋酸(24mg,0.40mmol)室温搅拌30分钟,随后加入三乙酰基硼氢化钠(128mg,0.61mmol),室温反应2h。加DCM和水萃取,有机层用食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物7-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬烷-2-羧酸叔丁酯(113.0mg,0.17mmol)。收率:85.0%。
LC/MS(ESI+)calcd for C36H47ClN7O4(M+H+)m/z,678.4;found,678.0.
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺的合成
将化合物叔丁基7-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬烷-2-羧酸叔丁酯(50mg,0.074mmol)溶于2mL CH2Cl2和2mL三氟乙酸,室温搅拌2h。溶剂旋干,加入2-(2,6-二氧代哌啶-3-基)-5-氟异吲哚-1,3-二酮(24mg,0.088mmol),DMSO 1mL,DIEA(95mg,0.737mmol),氮气保护,120℃搅拌反应过夜。TLC确定反应完全,反应液加入水和乙酸乙酯萃取3次,合并乙酸乙酯层,并用饱和食盐水萃洗3次,干燥旋干,柱层析分离得到黄色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺(17mg,0.020mmol)。收率:27%。
LC/MS(ESI+)calcd for C45H48ClN8O6(M+H+)m/z,834.3;found,834.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.60(d,J=8.2Hz,1H),7.86(d,J=8.8Hz,1H),7.80(d,J=9.6Hz,1H),7.63(d,J=8.3Hz,1H),7.39(d,J=2.3Hz,1H),7.33(d,J=9.7Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),6.77(s,1H),6.65(dd,J=8.5,1.7Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.53(t,J=7.4Hz,1H),4.47(d,J=13.3Hz,2H),3.85(d,J=8.0Hz,1H),3.74(s,4H),2.99(t,J=11.8Hz,2H),2.93–2.79(m,1H),2.63–2.51(m,2H),2.31(s,4H),2.11(s,4H),2.06–1.95(m,1H),1.84(dd,J=40.7,16.8Hz,9H),1.63(dd,J=23.7,10.8Hz,2H),1.50(dd,J=22.8,10.0Hz,2H),1.16–1.01(m,2H).
231:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2-,2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺的合成
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found,852.3.
1H NMR(400MHz,CDCl3)δ8.10(s,1H),7.97(d,J=9.3Hz,1H),7.88(d,J=8.1Hz,1H),7.56(d,J=8.7Hz,1H),7.36(d,J=10.8Hz,1H),7.00(d,J=2.4Hz,1H),6.98(d,J=9.8Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.81(d,J=7.5Hz,1H),4.91(dd,J=12.2,5.3Hz,1H),4.51(d,J=11.8Hz,2H),4.35–4.27(m,1H),4.05(dd,J=11.5,7.4Hz,1H),3.89(s,4H),3.04(s,2H),2.92–2.74(m,4H),2.37(s,3H),2.22–2.09(m,8H),1.86(s,4H),1.64(d,J=21.2Hz,4H),1.46(dd,J=20.7,8.5Hz,3H).
232:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)哒嗪-3-甲酰胺的合成
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found 851.8.
1H NMR(400MHz,CDCl3)δ8.11(s,1H),7.98(d,J=9.2Hz,1H),7.87(d,J=8.0Hz,1H),7.56(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.38(d,J=7.3Hz,1H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),6.59(d,J=9.3Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.32(t,J=9.8Hz,1H),4.05(d,J=8.1Hz,1H),3.93(s,3H),3.66(d,J=12.1Hz,2H),2.81(dtd,J=24.4,13.3,5.9Hz,5H),2.24–2.09(m,5H),1.96(s,5H),1.69(d,J=12.2Hz,10H),1.47(dd,J=22.5,10.3Hz,6H).
233:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found 851.8.
1H NMR(400MHz,CDCl3)δ8.82(s,1H),8.78–8.49(m,1H),7.95(s,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.8Hz,1H),7.38(d,J=6.6Hz,2H),6.99(d,J=2.0Hz,1H),6.85(dd,J=8.7,2.1Hz,1H),4.93(dd,J=12.1,5.3Hz,1H),4.45(d,J=13.2Hz,2H),4.32(d,J=10.0Hz,1H),4.03(d,J=7.6Hz,1H),3.24(s,2H),3.17(s,5H),3.03–2.92(m,2H),2.87(d,J=11.0Hz,1H),2.78(dd,J=23.8,12.5Hz,2H),2.53(s,2H),2.18(s,5H),2.05–1.84(m,8H),1.66(d,J=10.1Hz,2H),1.52–1.40(m,2H),1.26(t,J=10.7Hz,3H).
234:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((7-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-2-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found,852.3.
1H NMR(400MHz,CDCl3)δ8.82(d,J=1.1Hz,1H),8.43(s,1H),7.95(d,J=0.9Hz,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.42–7.33(m,2H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.46(d,J=13.2Hz,2H),4.35–4.24(m,1H),4.09–3.98(m,1H),3.16(d,J=5.2Hz,6H),3.03–2.87(m,3H),2.84–2.71(m,2H),2.55(s,2H),2.15(dd,J=14.9,4.7Hz,5H),2.06–1.85(m,6H),1.70(s,4H),1.53–1.38(m,3H),1.29(s,2H).
235:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮螺环[3.5]壬-2-基)吡嗪-2-甲酰胺的合成
LC/MS(ESI+)calcd for C44H47ClFN9O6(M+H+)m/z,852.3;found,852.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.57(d,J=1.1Hz,1H),8.12(d,J=8.2Hz,1H),7.86(d,J=8.8Hz,1H),7.79(d,J=1.0Hz,1H),7.70(d,J=11.4Hz,1H),7.43(d,J=7.4Hz,1H),7.37(d,J=2.4Hz,1H),7.12(dd,J=8.8,2.4Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.50(d,J=9.8Hz,1H),3.84(s,4H),3.60(d,J=11.8Hz,2H),3.31(s,1H),2.88(t,J=11.4Hz,3H),2.59(d,J=18.3Hz,1H),2.32(s,3H),2.20–2.00(m,5H),1.92–1.81(m,3H),1.77(s,6H),1.55(ddd,J=32.5,23.2,10.3Hz,5H),1.26(s,2H).
236:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-2-基)哌啶-1-基)吡嗪-2-甲酰胺的合成
LC/MS(ESI+)calcd for C44H45F4N9O6(M+H+)m/z,872.3;found,872.2.
1H NMR(400MHz,CDCl3)δ8.83(d,J=1.1Hz,1H),8.54(s,1H),7.96(d,J=1.0Hz,1H),7.74(d,J=8.6Hz,1H),7.47(d,J=10.9Hz,1H),7.39(dd,J=7.7,3.2Hz,2H),7.25(d,J=2.4Hz,1H),7.10(dd,J=8.6,2.4Hz,1H),4.93(dd,J=12.3,5.4Hz,1H),4.37(dd,J=22.3,11.9Hz,3H),4.04(dd,J=11.5,7.5Hz,1H),3.24(d,J=26.5Hz,3H),3.20–3.06(m,4H),2.94–2.67(m,3H),2.26–2.10(m,5H),1.99(s,4H),1.93–1.79(m,3H),1.55–1.40(m,4H).
237:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)哒嗪-3-甲酰胺的合成
1.化合物6-(2,5-二氮杂双环[2.2.1]庚基-2-基)-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺的合成
将化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(150mg,0.38mmol)、2,5-二氮杂双环[2.2.1]庚烷-2-羧酸叔丁酯(114mg,0.58mmol)、碳酸钾(265mg,1.92mmol)和2mL DMF依次加入反应器,氮气保护,80℃搅拌反应3h。TLC确定反应终点,反应液加入水,乙酸乙酯萃取3次,饱和食盐水萃洗3次,无水硫酸钠干燥,旋干得到粗品。硅胶柱层析分离得到类白色固体中间体。中间体加入1mL DCM溶清,随后加入1mL TFA,室温搅拌反应1h。TLC确定反应终点,反应液加入饱和碳酸氢钠水溶液,DCM萃取3次,无水硫酸钠干燥,旋干得到类白色化合物6-(2,5-二氮杂双环[2.2.1]庚基-2-基)-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(130mg,0.29mmol)。收率:75%。
LC/MS(ESI+)calcd for C23H25ClN6O2(M+H+)m/z,453.2;found,453.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)哒嗪-3-甲酰胺的合成
6-(2,5-二氮杂双环[2.2.1]庚基-2-基)-N-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(50mg,0.11mmol)加入(1R,5S,6r)-3-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-3-氮杂双环[3.1.0]己烷-6-甲醛(36mg,0.092mmol),2mLDCM/MeOH(1:1)溶清,加入冰醋酸(11mg,0.18mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(78mg,0.37mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)哒嗪-3-甲酰胺(68mg,0.083mmol)。收率:90%。LC/MS(ESI+)calcd for C42H41ClFN9O6(M+H+)m/z,822.3;found,822.0.
1H NMR(400MHz,CDCl3)δ8.74(d,J=32.3Hz,1H),8.02(d,J=9.3Hz,1H),7.89(t,J=11.6Hz,1H),7.55(t,J=9.3Hz,1H),7.38(d,J=12.5Hz,1H),7.06–6.96(m,2H),6.86(dd,J=8.8,2.4Hz,1H),6.73(d,J=9.4Hz,1H),4.94–4.86(m,1H),4.37–4.26(m,1H),4.12–3.94(m,2H),3.84(dd,J=8.2,4.7Hz,3H),3.53(d,J=6.2Hz,3H),3.28(s,1H),2.95–2.72(m,4H),2.67(s,2H),2.33(s,5H),2.14(dd,J=18.6,7.2Hz,6H),2.09–1.97(m,3H),1.47(dd,J=22.6,10.5Hz,2H).
238:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)吡嗪-3-甲酰胺的合成
LC/MS(ESI+)calcd for C42H41ClFN9O6(M+H+)m/z,822.3;found,822.0.
1H NMR(400MHz,CDCl3)δ8.95(d,J=56.0Hz,1H),8.83(d,J=0.9Hz,1H),7.70(s,1H),7.56(d,J=8.7Hz,1H),7.39(dd,J=10.3,4.7Hz,2H),7.39(dd,J=10.3,4.7Hz,2H),7.05(dd,J=7.3,4.6Hz,1H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),4.95–4.88(m,1H),4.36–4.24(m,1H),4.02(dd,J=11.4,7.5Hz,1H),3.85(dd,J=13.9,6.6Hz,3H),3.72(dd,J=14.0,7.0Hz,1H),3.57(t,J=9.4Hz,2H),3.48(d,J=9.4Hz,1H),3.19(s,1H),2.94–2.66(m,4H),2.59(s,2H),2.24–2.03(m,6H),1.93(d,J=8.5Hz,1H),1.71(d,J=9.7Hz,6H),1.47(dd,J=22.5,10.5Hz,2H).
239:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1r,5S)-8-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-3,8-二氮杂二环[3.2.1]辛基-3-基)哒嗪-3-甲酰胺的合成
LC/MS(ESI+)calcd for C43H43ClFN9O6(M+H+)m/z,836.3;found,836.2.
1H NMR(400MHz,CDCl3)δ8.42(s,1H),8.01(d,J=9.5Hz,1H),7.89(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.40(d,J=12.5Hz,1H),7.05(d,J=7.4Hz,1H),7.00(d,J=2.4Hz,1H),6.93–6.81(m,2H),4.92(dd,J=12.3,5.3Hz,1H),4.37–4.26(m,1H),4.13–3.95(m,3H),3.91(d,J=10.0Hz,2H),3.61(d,J=8.8Hz,4H),3.45(d,J=31.8Hz,2H),2.94–2.71(m,3H),2.53(s,2H),2.23–2.10(m,5H),2.04(s,2H),1.86–1.69(m,6H),1.53–1.39(m,3H),1.02(s,1H).
240:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((1r,5S)-8-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-3,8-二氮杂二环[3.2.1]辛基-3-基)吡嗪-3-甲酰胺的合成
LC/MS(ESI+)calcd for C43H43ClFN9O6(M+H+)m/z,836.3;found,836.2.
1H NMR(400MHz,CDCl3)δ8.85(d,J=1.1Hz,1H),8.50(s,1H),7.87(d,J=0.9Hz,1H),7.56(d,J=8.7Hz,1H),7.42–7.36(m,2H),7.05(d,J=7.4Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),4.92(dd,J=12.3,5.3Hz,1H),4.34–4.26(m,1H),4.09–4.01(m,1H),4.01–3.93(m,2H),3.93–3.86(m,2H),3.60(d,J=9.6Hz,2H),3.58–3.50(m,2H),3.36(s,2H),2.94–2.84(m,1H),2.83–2.71(m,2H),2.50(s,2H),2.16(ddd,J=19.5,8.5,5.2Hz,5H),2.00(s,2H),1.70(s,6H),1.47(dd,J=22.6,10.5Hz,2H),1.00(s,1H).
241:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2-,2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)吡嗪-3-羧酰胺的合成
1.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)吡嗪-3-甲酰胺
化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)吡嗪-3-甲酰胺(300mg,0.77mmol),4-羟甲基哌啶(265mg,2.30mmol),碳酸钾(530mg,3.83mmol)和DMF 5mL依次加入反应器,氮气保护,80℃反应3小时,TLC确定反应终点。确认反应完毕后,反应液加入水和乙酸乙酯萃取3次,合并乙酸乙酯层,并用饱和食盐水萃洗3次,干燥旋干,柱层析分离得到白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)吡嗪-3-甲酰胺(355mg,0.76mmol),收率:98%。
LC/MS(ESI+)calcd for C24H27ClN5O3(M+H+)m/z,470.2;found,470.2.
2.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)六氢环戊[c]吡咯-2(1H)-基)吡嗪-3-甲酰胺的合成
将化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)吡嗪-3-甲酰胺(70mg,0.15mmol)加入2mL DCM溶清,加入DMP(76mg,0.18mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(六氢吡咯[3,4-c]吡咯-2(1H)-基)异吲哚-1,3-二酮(68mg,0.18mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(18mg,0.30mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(126mg,0.60mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)甲基)六氢环戊[c]吡咯-2(1H)-基)吡嗪-3-甲酰胺(86mg,0.10mmol)。收率:69%。
LC/MS(ESI+)calcd for C45H48ClN8O6(M+H+)m/z,834.3;found,834.3.
1H NMR(400MHz,CDCl3)δ8.79(s,1H),7.98(s,1H),7.64(d,J=8.2Hz,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=8.1Hz,1H),7.01(d,J=2.0Hz,1H),6.86(dd,J=8.7,2.0Hz,1H),6.78(s,1H),6.52(d,J=8.3Hz,1H),4.93(dd,J=12.0,5.2Hz,1H),4.93(dd,J=12.0,5.2Hz,1H),4.46(d,J=12.8Hz,2H),4.32(t,J=9.9Hz,1H),4.01(d,J=7.6Hz,1H),3.75(s,4H),3.08–2.96(m,3H),2.82–2.75(m,3H),2.40(s,4H),2.15(dd,J=25.3,8.7Hz,7H),1.91(d,J=22.6Hz,6H),1.69(dd,J=22.2,10.6Hz,2H),1.48(dd,J=22.2,10.9Hz,2H),1.23(d,J=18.1Hz,2H).
242:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺[3.5]壬-2-基)吡嗪-2-羧酰胺的合成
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H+)m/z,838.3;found 838.3.
1H NMR(400MHz,CDCl3)δ8.83(d,J=1.2Hz,1H),8.24(s,1H),7.59(d,J=1.1Hz,1H),7.56(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.39(d,J=7.2Hz,2H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.35–4.25(m,1H),4.08–3.98(m,1H),3.90(s,4H),3.73(d,J=11.9Hz,2H),2.95–2.83(m,4H),2.82–2.71(m,2H),2.66(d,J=30.0Hz,4H),2.23–2.10(m,5H),1.94(s,6H),1.84–1.61(m,6H),1.46(dd,J=22.8,10.5Hz,2H).
243:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺的合成
1.化合物5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-羧酸乙酯的合成
化合物5-氯-1,3,4-噻二唑-2-羧酸乙酯(200mg,1.04mmol),4-羟甲基哌啶(179mg,1.56mmol),碳酸钾(287mg,2.08mmol)和DMF 5mL依次加入反应器,氮气保护,60℃反应1h。TLC确定反应终点,反应液加入水和乙酸乙酯萃取3次,并用饱和食盐水萃洗2次,干燥旋干,pre-TLC分离得到白色固体化合物5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-羧酸乙酯(167mg,0.61mmol),收率:59%。LC/MS(ESI+)calcd for C11H17N3O3S(M+H+)m/z,272.1;found,272.1.2.N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺
化合物5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-羧酸乙酯(80mg,0.29mmol),4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(110mg,0.44mmol)和无水甲醇1mL依次加入反应器,回流反应2d。TLC确认反应终点。反应完毕,蒸除溶剂,pre-TLC纯化得到淡黄色固体N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺(88mg,0.18mmol),收率:63%;LC/MS(ESI+)calcd for C22H26ClN5O3S(M+H+)m/z,476.1;found,476.1.
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺的合成
化合物N-(1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺(44mg,0.92mmol)加入2mL DCM溶清,加入DMP(47mg,0.11mmol)室温搅拌反应2h。TLC确定反应终点,反应液过滤滤饼用DCM淋洗,滤液旋干得到粗品。粗品加入2-(2,6-二氧哌啶-3-基)-4-(哌嗪-1-基)异吲哚-1,3-二酮(41mg,0.10mmol),2mL DCM/MeOH(1:1)溶清,加入冰醋酸(11mg,0.18mmol)室温搅拌反应30min,随后加入三乙酰基硼氢化钠(78mg,0.37mmol)室温搅拌反应2h。TLC确定反应终点,加入饱和碳酸氢钠水溶液,二氯甲烷萃取3次合并干燥旋干,pre-TLC分离得到类白色固体化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺(50mg,0.058mmol)。收率:63%。
LC/MS(ESI+)calcd for C42H46ClN9O6S(M+H+)m/z,840.3;found,840.3.
1H NMR(400MHz,CDCl3)δ8.08(s,1H),7.64(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.00(t,J=5.0Hz,2H),6.84(dd,J=8.7,2.4Hz,1H),6.78(d,J=1.9Hz,1H),6.51(d,J=8.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.36–4.26(m,1H),4.00(dd,J=23.9,12.2Hz,3H),3.74(s,4H),3.19(t,J=11.9Hz,2H),2.94–2.73(m,3H),2.38(s,3H),2.15(ddd,J=17.9,11.1,5.1Hz,7H),1.85(s,6H),1.70–1.57(m,4H),1.53–1.40(m,2H),1.32(d,J=13.0Hz,2H).
244:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-噻二唑-2-甲酰胺的合成
LC/MS(ESI+)calcd for C42H46ClFN9O6S(M+H+)m/z,858.3;found 858.3.
1H NMR(400MHz,CDCl3)δ7.61–7.52(m,1H),7.36(d,J=10.9Hz,1H),7.01(d,J=2.4Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.81(d,J=7.5Hz,1H),4.91(dd,J=12.1,5.6Hz,1H),4.33(t,J=9.9Hz,1H),3.99(dd,J=19.1,11.8Hz,3H),3.90(d,J=1.9Hz,4H),3.20(dd,J=12.6,10.3Hz,2H),2.44(s,4H),2.26(d,J=6.9Hz,2H),2.23–2.10(m,5H),2.05(s,1H),1.98–1.79(m,7H),1.66(dt,J=20.0,10.1Hz,2H),1.50(dd,J=22.0,11.4Hz,2H),1.39–1.29(m,2H
245:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-吡唑-4-羧酰胺
第一步:将1H-吡唑-4-羧酸乙酯(1.4g,10mmol),氢氧化钠(0.8g,20mmol)依次加入到10mL的水中中,加热至回流反应2小时,TLC监测反应完全后,自然冷却至室温,用6N盐酸调节PH值到4-5左右,过滤得1.05g化合物1H-吡唑-4-甲酸,收率:93%。
第二步:将1H-吡唑-4-甲酸(336mg,3mmol)溶于10mL DMF中,加入DIEA(774mg,6mmol),加入HATU(1.36g,3.6mmol),室温下反应半小时。加入4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(750mg,3mmol)反应6小时,TLC监测反应完全后,5mL饱和氯化铵淬灭反应,10mL乙酸乙酯萃取,水层反萃一次,合并有机层,用饱和食盐水洗涤,无水硫酸钠干燥,浓缩,色谱柱纯化,得490mg化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-吡唑-4-甲酰胺,收率48%。
第三步:将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1H-吡唑-4-甲酰胺(344mg,1mmol),(1s,4s)-4-((甲磺酰基)氧基)环己烷-1-羧酸甲酯(708mg,3mmol),溶于4mL DMF中,加入碳酸钾(414mg,3mmol),80°下反应过夜。5mL饱和碳酸氢钠淬灭反应,10mL乙酸乙酯萃取,水层反萃一次,合并有机层,用饱和食盐水洗涤,无水硫酸钠干燥,浓缩,色谱柱纯化,得144mg化合物(1R,4r)-4-(4-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-吡唑-1-基)环己烷-1-羧酸甲酯,收率:30%。
第四步:将(1R,4r)-4-(4-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)-1H-吡唑-1-基)环己烷-1-羧酸甲酯(144mg,0.3mmol)溶于10mL甲醇,在冰浴下加入硼氢化钠(58mg,1.5mmol),加毕后缓慢升至室温,60℃下搅拌过夜,5mL饱和氯化铵淬灭反应,10ml乙酸乙酯萃取,有机层减压浓缩,薄层色谱层析分离纯化得106mg化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-吡唑-4-甲酰胺,收率:77%。
第五步:将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-(羟甲基)环己基)-1H-吡唑-4-甲酰胺(106mg,0.23mmol),溶于4mL二氯甲烷,加入DMP(127mg,0.3mmol),室温反应过夜。过滤,浓缩,色谱柱纯化,得88mg化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-吡唑-4-甲酰胺,收率:85%。
第六步:将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-吡唑-4-甲酰胺(45mg,0.1mmol)溶于3mL二氯甲烷,加入2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(76mg,0.2mmol),加入冰乙酸(12mg,0.02mmol),室温搅拌30min,加入三乙酰基硼氢化钠(84.8mg,0.4mmol),室温反应2小时。5mL饱和碳酸氢钠淬灭反应,10ml乙酸乙酯萃取,有机层减压浓缩,薄层色谱层析分离纯化得30mg化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-吡唑-4-羧酰胺,收率:37%。1H NMR(400MHz,CDCl3)δ7.91(s,1H),7.70(d,J=5.6Hz,1H),7.64(d,J=8.0Hz,1H),7.55(d,J=8.8Hz,1H),6.98(d,J=2.4Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),6.77(d,J=2.0Hz,1H),6.50(dd,J=8.8,2.0Hz,1H),5.60(d,J=8.0Hz,1H),4.93(dd,J=12.0,5.2Hz,1H),4.16(m,4H),3.73(s,4H),3.02(s,1H),2.81(m,4H),2.37(s,4H),2.17(m,8H),2.01(m,2H),1.86(m,4H),1.40(m,3H),1.09(m,3H).LC/MS(ESI+)calcd for C44H49ClN8O6([M+H]+)m/z:821.4;found 821.4。
246:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-吡唑-4-羧酰胺
将N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-甲酰环己基)-1H-吡唑-4-甲酰胺(38mg,0.08mmol)溶于3mL二氯甲烷,加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚-1,3-二酮(40mg,0.1mmol),加入冰乙酸(6mg,0.01mmol),室温搅拌30min,加入三乙酰基硼氢化钠(53mg,0.25mmol),室温反应2小时。5mL饱和碳酸氢钠淬灭反应,10ml乙酸乙酯萃取,有机层减压浓缩,薄层色谱层析分离纯化得24mg化合物N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-吡唑-4-羧酰胺,收率:36%。1H NMR(400MHz,CDCl3)δ8.24(s,1H),7.94(s,1H),7.72(d,J=5.6Hz,1H),7.55(d,J=8.8Hz,1H),7.35(d,J=7.6Hz,1H),6.98(d,J=2.4Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),6.80(d,J=7.6Hz,1H),5.67(d,J=8.0Hz,1H),4.91(dd,J=12.0,5.6Hz,1H),4.27(m,1H),4.04(m,2H),3.89(s,4H),2.80(m,8H),1.67(m,8H),1.31(m,15H).LC/MS(ESI+)calcd for C44H48ClFN8O6([M+H]+)m/z:839.4;found 839.4。
247:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-4-氧代-3,4-二氢喹唑啉-7-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
第一步:将3-(7-氟-4-氧喹唑啉-3(4H)-基)哌啶-2,6-二酮(275mg,1mmol),2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯(226mg,1mmol),溶于4mL DMSO中,加入DIPEA(258mg,2mmol),在N2保护下130°下反应2小时。5mL饱和氯化铵淬灭反应,10mL乙酸乙酯萃取,水层反萃一次,合并有机层,用饱和食盐水洗涤,无水硫酸钠干燥,浓缩,色谱柱纯化,得112mg化合物2-(3-(2,6-二氧哌啶-3-基)-4-氧代-3,4-二氢喹唑啉-7-基)-2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯,收率:23%。
第二步:将2-(3-(2,6-二氧哌啶-3-基)-4-氧代-3,4-二氢喹唑啉-7-基)-2,7-二氮杂螺[3.5]壬烷-7-羧酸叔丁酯(112mg,0.23mmol),溶于5mL二氯甲烷中,加入5mL三氟乙酸,在室温下反应2小时。减压浓缩除去溶剂和三氟乙酸,加入5mL二氯甲烷,加入0.5mL饱和碳酸氢钠,多次5mL二氯甲烷萃取,合并有机层,无水硫酸钠干燥,浓缩,色谱柱纯化,得46mg化合物3-(4-氧代-7-(2,7-二氮杂螺[3.5]壬-2-基)喹唑啉-3(4H)-基)哌啶-2,6-二酮,收率:53%。
第三步:将3-(4-氧代-7-(2,7-二氮杂螺[3.5]壬-2-基)喹唑啉-3(4H)-基)哌啶-2,6-二酮(46mg,0.12mmol)溶于3mL二氯甲烷,加入N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-甲酰哌啶-1-基)哒嗪-3-甲酰胺(93mg,0.2mmol),加入冰乙酸(12mg,0.02mmol),室温搅拌30min,加入三乙酰基硼氢化钠(51mg,0.24mmol),室温反应2小时。5mL饱和碳酸氢钠淬灭反应,10ml乙酸乙酯萃取,有机层减压浓缩,薄层色谱层析分离纯化得21mg化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2,6-二氧哌啶-3-基)-4-氧代-3,4-二氢喹唑啉-7-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺,收率:21%。1H NMR(400MHz,CDCl3)δ8.59(s,1H),8.05(d,J=8.8Hz,1H),7.96(d,J=9.2Hz,1H),7.89(d,J=8.0Hz,1H),7.55(d,J=8.8Hz,1H),6.98(m,2H),6.84(dd,J=8.4,2.4Hz,1H),6.55(dd,J=7.6,2.4Hz,1H),6.50(d,J=2.4Hz,1H),5.14(d,J=2.8Hz,1H),4.53(m,5H),4.31(m,1H),4.05(m,1H),3.73(s,4H),3.48(s,2H),3.03(t,J=12.4Hz,2H),2.92(m,1H),2.77(m,2H),2.21(m,7H),1.69(m,4H),1.44(m,4H),0.85(m,4H).LC/MS(ESI+)calcd for C44H49ClN10O5([M+H]+)m/z:834.4;found834.4。
248:2-氯-4-((1R,3R)-3-(5-(4-((4-(2-(2,6-二氧哌啶-3-基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.合成中间体24-1:2-氯-4-((1R,3R)-3-(5-(4-(羟甲基)哌啶-1-基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物4-((1R,3R)-3-(5-溴-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(1-1)(189mg,0.4mmol)和哌啶-4-基甲醇(69mg,0.6mmol)溶于3mL甲苯中,加入三乙胺1mL,Pd(dppf)2Cl2(30mg)和CuI(150mg),N2条件保护下,反应液温度升高到90-100℃,继续搅拌反应12h,反应完成后,经减压蒸馏除去溶剂。粗品采用板层析纯化(PE:EA=1:1),得到163mg中间体24-1,收率:80%。质谱(ES):m/z=508[M+H]+。
2.(1-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌啶-4-基)甲基甲磺酸盐
将上一步得到的中间体(508mg,1mmol)溶于10mLDCM中,加入TEA(152mg,1.5mmol),冰浴条件下滴加甲基磺酰氯(137mg,1.2mmol),搅拌反应3h。反应完成后,加入5mL水和20mL二氯甲烷萃取,得到的有机相,经减压蒸馏除去溶剂,粗品未进纯化直接用于下一步反应。
3.2-氯-4-((1R,3R)-3-(5-(4-((4-(2-(2,6-二氧哌啶-3-基)-1-氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物(1-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异辛醇-5-基)哌啶-4-基)甲基甲磺酸盐(中间体24-2)(76mg,0.13mmol)溶于2mL DMSO中,加入3-(1-氧代-5-(哌嗪-1-基)异吲哚啉-2-基)哌啶-2,6-二酮(43mg,0.13mmol)后。反应液温度升高到80℃,继续搅拌2h反应完成。反应完成后,将反应液温度降到室温,加入10mL水和15mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂。粗品采用板层析纯化(DCM:MeOH=10:1),得到62mg最终产物,收率:58%。质谱(ES):m/z=818[M+H]+。
1H NMR(400MHz,DMSO-d6)δ10.95(s,1H),7.91(d,J=8.7Hz,1H),7.53(d,J=8.4Hz,1H),7.47(d,J=9.1Hz,1H),7.29(d,J=2.2Hz,1H),7.15-6.99(m,5H),5.11–5.01(m,1H),4.67(s,2H),4.53(s,1H),4.28(t,J=16.7Hz,3H),3.87(d,J=11.9Hz,2H),3.44(s,3H),2.85(d,J=11.5Hz,3H),2.72-2.55(m,1H),2.45-2.29(m,2H),2.22(d,J=6.5Hz,2H),2.04-1.74(m,6H),1.39(s,6H),1.22(d,J=17.3Hz,4H),1.14(s,6H).
249:2-氯-4-((1R,3R)-3-(5-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物(1-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异吲哚啉-5-基)哌啶-4-基)甲基甲磺酸盐(中间体24-2)(76mg,0.13mmol)溶于2mLDMSO中,加入2-(2,6-二氧哌啶-3-基)-5-(哌嗪-1-基)异吲哚啉-1,3-二酮(45mg,0.13mmol)后。反应液温度升高到80℃,继续搅拌2h反应完成。反应完成后,将反应液温度降到室温,加入10mL水和15mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂。粗品采用板层析纯化(DCM:MeOH=10:1),得到61 mg最终产物,收率:56%。质谱(ES):m/z=832[M+H]+。
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),7.91(d,J=8.7Hz,1H),7.69(d,J=8.4Hz,1H),7.47(d,J=9.0Hz,1H),7.30(t,J=19.4Hz,3H),7.11-7.00(m,3H),5.14-5.04(m,1H),4.67(s,2H),4.53(s,1H),4.29(s,1H),3.87(d,J=12.0Hz,2H),3.45(s,3H),2.84(t,J=11.7Hz,3H),2.58(d,J=25.3Hz,4H),2.22(d,J=5.9Hz,2H),2.00(s,2H),1.82(d,J=9.9Hz,3H),1.39(s,6H),1.30-1.17(m,4H),1.16(d,J=15.4Hz,6H).
250:2-氯-4-((1r,3r)-3-(5-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-3-氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物(1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异辛醇-5-基)哌啶-4-基)甲基甲磺酸盐(76mg,0.13mmol)溶于2mL DMSO中,加入3-(5-氟-1-氧代-6-(哌嗪-1-基)异丙醇-2-基)哌啶-2,6-二酮(45mg,0.13mmol)后。反应液温度升高到80℃,继续搅拌2h反应完成。反应完成后,将反应液温度降到室温,加入10mL水和15mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂。粗品采用板层析纯化(DCM:MeOH=10:1),得到63mg最终产物,收率:58%。质谱(ES):m/z=836[M+H]+。
1H NMR(400MHz,DMSO-d6)δ11.03(s,1H),7.93-7.89(m,1H),7.46(s,3H),7.29(s,2H),7.05(s,3H),5.16-5.03(m,2H),4.68(s,2H),4.53(s,2H),4.42-4.34(m,2H),4.29(s,2H),3.93-3.86(m,2H),2.94-2.78(m,4H),2.25-2.21(m,2H),2.03-1.96(m,3H),1.88-1.76(m,4H),1.39(s,6H),1.24(s,4H),1.14(s,6H).
251:2-氯-4-((1r,3r)-3-(5-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧代异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-1-氧代异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将化合物(1-(2-((1R,3R)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧代异辛醇-5-基)哌啶-4-基)甲基甲磺酸盐(76mg,0.13mmol)溶于2mL DMSO中,加入3-(6-氟-1-氧-5-(哌嗪-1-基)异内酰胺-2-基)哌啶-2,6-二酮(45mg,0.13mmol)后。反应液温度升高到80℃,继续搅拌2h反应完成。反应完成后,将反应液温度降到室温,加入10mL水和15mL乙酸乙酯萃取,得到的有机相,经减压蒸馏除去溶剂。粗品采用板层析纯化(DCM:MeOH=10:1),得到65mg最终产物,收率:58%。质谱(ES):m/z=836[M+H]+。
1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),7.91(d,J=8.8Hz,1H),7.45(dd,J=16.3,10.3Hz,2H),7.26(dd,J=16.5,4.8Hz,2H),7.07(d,J=11.7Hz,3H),5.12-5.02(m,1H),4.67(s,2H),4.53(s,1H),4.35(s,1H),4.28(d,J=4.7Hz,2H),3.87(d,J=11.9Hz,2H),3.13(s,3H),2.85(d,J=13.3Hz,3H),2.55(s,3H),2.42-2.31(m,1H),2.24(d,J=6.2Hz,2H),1.99(s,2H),1.82(d,J=10.9Hz,3H),1.37(d,J=11.8Hz,6H),1.25(d,J=9.9Hz,4H),1.14(s,6H).
252:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
253:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((5-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
1.4-(((甲基磺酰基)氧基)甲基)哌啶-1-羧酸苄基酯的合成
化合物4-(羟甲基)哌啶-1-羧酸苄酯(2.00g,8.00mmol)加入到20ml CH2Cl2中,冰浴下搅拌,再加入TEA(2.5mL,16.00mmol)和MsCl(1.10g,7.30mmol),加完后撤掉冰浴,常温反应4h。反应液用DCM稀释,分别用1N盐酸洗涤,饱和NaHCO3溶液洗涤和饱和NaCl溶液洗涤,加入无水硫酸钠干燥,旋干,得无色油状化合物4-(((甲基磺酰基)氧基)甲基)哌啶-1-羧酸苄基酯(2.50g,7.60mmol),收率:96%。
LC/MS(ESI+)calcd for C15H20NO5S(M+H+)m/z,328.2;found,328.2.
2.化合物5-((1-((苄氧基)羰基)哌啶-4-基)甲基)六氢吡咯并[3,4-c]吡咯-2(1H)-羧酸叔丁酯
化合物4-(((甲基磺酰基)氧基)甲基)哌啶-1-羧酸苄基酯(2.00g,6.10mmol),2-BOC-八氢吡咯基[3,4-c]吡咯(1.30g,6.10mmol),K2CO3(2.10g,15.00mmol)和KI(199mg,1.20mmol),加入到CH3CN中,加热到85℃,反应过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层再用饱和食盐水洗涤,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到无色油状化合物5-((1-((苄氧基)羰基)哌啶-4-基)甲基)六氢吡咯并[3,4-c]吡咯-2(1H)-羧酸叔丁酯(1.60g,3.60mmol),收率:60%。LC/MS(ESI+)calcdfor:C25H36N3O4(M+H+)m/z,444.2;found,444.2.
3.化合物5-(哌啶-4-基甲基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸叔丁酯的合成
化合物5-((1-((苄氧基)羰基)哌啶-4-基)甲基)六氢吡咯并[3,4-c]吡咯-2(1H)-羧酸叔丁酯(1.60g,3.60mmol)溶于MeOH,Ar保护下抽气三次,加入Pd/C,室温反应过夜。硅藻土过滤,浓缩得到灰色油状化合物5-(哌啶-4-基甲基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸叔丁酯(1.00g,3.20mmol),收率:90%。LC/MS(ESI+)calcdfor:C17H30N3O2(M+H+)m/z,400.2;found,400.2.
4.化合物5-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)六氢吡咯烷酮并[3,4-c]吡咯-2(1H)-羧酸叔丁酯的合成
化合物5-(哌啶-4-基甲基)六氢吡咯[3,4-c]吡咯-2(1H)-羧酸叔丁酯(98mg,0.30mmol),6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(117mg,0.30mmol)和TEA(60mg,0.60mmol)加入到3mL 1,4-dioxane中。加热至110℃,反应过夜。冷却至室温,加入水和乙酸乙酯萃取,有机层再用饱和食盐水洗涤,加入无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到化合物5-((1-(6-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基甲酰基)哒嗪-3-基)哌啶-4-基)甲基)六氢吡咯烷酮并[3,4-c]吡咯-2(1H)-羧酸叔丁酯(70mg,0.10mmol),收率:35%。
LC/MS(ESI+)calcdfor:C35H45ClN7O4(M+H+)m/z,664.3;found,664.3.
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(70mg,0.10mmol)加入到1mL CH2Cl2中,冰浴下搅拌,再缓慢滴加2mL TFA,加完后撤去冰浴,常温反应2h。溶剂旋干,得到化合物N-((1R,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(53mg,0.09mmol),收率:95%。
LC/MS(ESI+)calcdfor:C30H37ClN7O2(M+H+)m/z,564.2;found,564.2.
6.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺的合成
化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(53mg,0.09mmol),2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(24mg,0.09mmol)和DIEA(34mg,0.27mmol)加入到DMSO中,加热至130℃,搅拌2.5h。冷却至室温,加水和乙酸乙酯萃取,有机层用食盐水洗涤,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺252(26mg,0.03mg),收率36%。LC/MS(ESI+)calcd for:C43H45ClN9O6(M+H+)m/z,820.3;found,820.3.1H NMR(400MHz,CDCl3)δ8.18(s,1H),7.98(d,J=9.4Hz,1H),7.88(d,J=8.2Hz,1H),7.70(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.07-6.94(m,3H),6.87(dd,J=8.8,2.4Hz,1H),4.96(m,1H),4.44(m,6H),4.07(d,J=10.8Hz,1H),3.69(d,J=26.3Hz,3H),3.39(s,2H),3.12-2.60(m,9H),2.28-2.11(m,6H),1.54-1.42(m,4H),1.37(d,J=15.3Hz,2H),0.90(t,J=6.5Hz,2H).7.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((5-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
得到目标化合物253,收率42%。LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.2;found,838.2.1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.59(d,J=8.2Hz,1H),7.85(d,J=8.8Hz,1H),7.78(d,J=9.5Hz,1H),7.64(d,J=12.5Hz,1H),7.38(d,J=2.4Hz,1H),7.30(d,J=9.7Hz,1H),7.20-7.01(m,2H),5.08(dd,J=12.8,5.4Hz,1H),4.49(dd,J=28.6,8.6Hz,3H),3.91-3.65(m,3H),3.04-2.78(m,5H),2.64-2.56(m,1H),2.50(m,J=1.8Hz,5H),2.25(d,J=6.6Hz,2H),2.15-1.97(m,4H),1.84(dd,J=31.4,11.7Hz,5H),1.69-1.43(m,4H),1.14(dd,J=47.4,16.7Hz,3H).
254:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcdfor:C43H46ClN9O6(M+H+)m/z,820.3;found 820.3.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.56(d,J=1.2Hz,1H),8.20(s,1H),8.07(d,J=8.2Hz,1H),7.85(d,J=8.8Hz,1H),7.65(d,J=8.4Hz,1H),7.36(d,J=2.4Hz,1H),7.11(dd,J=8.8,2.4Hz,1H),6.95(d,J=2.1Hz,1H),6.86(dd,J=8.6,2.1Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),4.50(td,J=9.9,4.4Hz,1H),4.42(d,J=13.0Hz,2H),3.80(dd,J=7.8,3.8Hz,1H),3.73-3.59(m,2H),3.01-2.79(m,6H),2.25(d,J=6.6Hz,2H),2.04-1.94(m,2H),1.91-1.68(m,6H),1.66-1.42(m,6H),1.34(d,J=5.8Hz,1H),1.27-1.17(m,5H),1.06(q,J=12.9Hz,2H),0.84(t,J=6.6Hz,1H).
255:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.2;found 838.2.
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.56(d,J=1.2Hz,1H),8.21(d,J=1.4Hz,1H),8.07(d,J=8.3Hz,1H),7.85(d,J=8.8Hz,1H),7.63(d,J=12.5Hz,1H),7.36(d,J=2.4Hz,1H),7.17-7.06(m,2H),5.08(dd,J=12.9,5.4Hz,1H),4.55-4.38(m,3H),3.76(dt,J=34.4,9.7Hz,3H),3.01-2.80(m,5H),2.57(s,1H),2.54(s,1H),2.50(p,J=1.8Hz,5H),2.25(d,J=6.7Hz,2H),2.13-1.97(m,3H),1.91-1.72(m,5H),1.67-1.40(m,4H),1.23(d,J=8.4Hz,1H),1.14-0.97(m,2H).
256:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)嘧啶-5-羧酰胺
LC/MS(ESI+)calcdfor:C43H46ClN9O6(M+H+)m/z,820.3;found 820.3.
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.71(s,2H),8.10(d,J=7.5Hz,1H),7.85(d,J=8.7Hz,1H),7.65(d,J=8.4Hz,1H),7.37(d,J=2.4Hz,1H),7.12(dd,J=8.8,2.5Hz,1H),6.95(d,J=2.1Hz,1H),6.85(dd,J=8.6,2.1Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),4.68(d,J=12.9Hz,2H),4.59-4.44(m,1H),3.84-3.59(m,3H),3.01-2.78(m,5H),2.23(d,J=6.7Hz,2H),2.14-1.95(m,4H),1.90(t,J=9.3Hz,2H),1.77(d,J=12.6Hz,3H),1.48(p,J=5.4,4.7Hz,5H),1.33(d,J=5.8Hz,1H),1.27-1.17(m,4H),0.98(d,J=12.6Hz,2H),0.83(d,J=7.1Hz,1H).
257:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((5-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)嘧啶-5-羧酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.2;found 838.2.
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.72(s,2H),8.10(d,J=7.5Hz,1H),7.85(d,J=8.8Hz,1H),7.63(d,J=12.5Hz,1H),7.38(d,J=2.4Hz,1H),7.32-6.97(m,2H),5.08(dd,J=12.8,5.4Hz,1H),4.69(d,J=12.7Hz,2H),4.53(d,J=4.6Hz,1H),3.74(d,J=29.2Hz,3H),3.00-2.80(m,6H),2.60(d,J=3.3Hz,1H),2.50(m,6H),2.25(d,J=6.6Hz,2H),2.09(s,2H),1.95-1.85(m,2H),1.78(d,J=12.5Hz,3H),1.57-1.42(m,4H),1.23(d,J=7.9Hz,1H),1.00(d,J=12.6Hz,2H).
258:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((5-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)甲基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcdfor:C45H48ClN7O6(M+H+)m/z,818.3;found,818.3.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),7.95(d,J=7.6Hz,1H),7.85(d,J=8.7Hz,1H),7.68(dd,J=19.6,8.4Hz,2H),7.65(d,J=8.4Hz,1H),7.38(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.5Hz,1H),7.01-6.76(m,4H),5.06(dd,J=12.9,5.4Hz,1H),4.57-4.43(m,1H),3.86-3.58(m,5H),3.30-3.19(m,2H),3.02-2.81(m,2H),2.71(t,J=12.1Hz,2H),2.57(dd,J=19.0,7.2Hz,4H),2.25(d,J=7.2Hz,2H),2.13-2.06(m,2H),2.04-1.96(m,2H),1.93-1.84(m,2H),1.75(d,J=12.6Hz,2H),1.69-1.58(m,1H),1.49(q,J=12.8Hz,4H),1.23(d,J=7.4Hz,2H),1.14(m,6.8Hz,2H).
259:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1-(2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)哒嗪-3-羧酰胺
LC/MS(ESI+)calcdfor:C43H46ClN9O6(M+H+)m/z,820.3;found 820.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.2Hz,1H),7.84(dd,J=10.5,9.1Hz,2H),7.63(d,J=8.6Hz,1H),7.39(d,J=2.4Hz,1H),7.28(d,J=2.3Hz,1H),7.19(dd,J=8.7,2.3Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),7.00(d,J=9.4Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),4.53(dt,J=10.3,5.8Hz,1H),4.01(dd,J=10.2,6.8Hz,2H),3.90-3.71(m,3H),3.40(d,J=10.3Hz,2H),3.01-2.80(m,5H),2.62-2.52(m,4H),2.24(d,J=6.6Hz,2H),2.15-2.05(m,2H),2.00(m,1H),1.94-1.85(m,2H),1.81-1.42(m,8H),1.22(s,1H),1.11(d,J=11.7Hz,2H).
260:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(5-((1-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)哌啶-4-基)甲基)六氢吡咯并[3,4-c]吡咯-2(1H)-基)哒嗪-3-羧酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.3;found 838.3.
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.57(d,J=8.2Hz,1H),7.85(dd,J=11.7,9.0Hz,2H),7.69(d,J=11.5Hz,1H),7.45-7.36(m,2H),7.14(dd,J=8.8,2.4Hz,1H),7.01(d,J=9.5Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.54(td,J=10.3,5.1Hz,1H),3.82(m,3H),3.57(d,J=12.1Hz,2H),3.46-3.39(m,2H),3.04-2.78(m,5H),2.63-2.55(m,3H),2.29(d,J=7.1Hz,2H),2.21-1.96(m,4H),1.95-1.86(m,2H),1.85-1.74(m,2H),1.72-1.58(m,3H),1.51(q,J=11.5Hz,2H),1.23(t,J=6.2Hz,4H).
261:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(9-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-3,9-二氮杂螺[5.5]十一烷-3-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcdfor:C46H51ClFN9O6(M+H+)m/z,880.3;found 880.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.61(d,J=8.2Hz,1H),7.83(dd,J=22.8,9.0Hz,2H),7.70(d,J=11.3Hz,1H),7.46-7.36(m,2H),7.32(d,J=9.7Hz,1H),7.13(dd,J=8.8,2.5Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.51(d,J=11.4Hz,1H),3.85(d,J=10.4Hz,1H),3.76-3.52(m,4H),3.58(t,J=9.2Hz,2H),2.87(t,J=11.9Hz,3H),2.34(s,4H),2.24-1.96(m,6H),1.94-1.76(m,5H),1.50(d,J=8.2Hz,10H),1.31-1.11(m,5H).
262:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(9-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-3,9-二氮杂螺[5.5]十一烷-3-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcdfor:C46H52ClN9O6(M+H+)m/z862.3;found 862.3.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.61(d,J=8.2Hz,1H),7.83(dd,J=23.3,9.1Hz,2H),7.65(d,J=8.5Hz,1H),7.39(d,J=2.4Hz,1H),7.31(d,J=10.3Hz,2H),7.22(d,J=8.7Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),4.53(s,1H),4.03(d,J=13.1Hz,2H),3.91-3.80(m,1H),3.68(d,J=6.3Hz,4H),2.90(dq,J=31.6,9.1,5.8Hz,3H),2.34(s,4H),2.12(dd,J=19.3,9.1Hz,4H),2.00(d,J=13.5Hz,2H),1.93-1.73(m,5H),1.50(d,J=6.8Hz,10H),1.29-1.06(m,5H).
263:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((2R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2-甲基哌嗪-1-基)哒嗪-3-甲酰胺
1.叔丁基(R)-4-(6-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)-3-甲基哌嗪-1-羧酸盐的合成
化合物6-氯-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(200mg,0.51mmol),叔丁基(R)-3-甲基哌嗪-1-羧酸酯(200mg,1.00mmol),K2CO3(207mg,1.50mmol),加入到5mL NMP中,120℃反应6h。用大量EA稀释反应液,分别用水和饱和NaCl溶液洗涤有机相,加入无水硫酸钠干燥,旋干,柱层析纯化得化合物叔丁基(R)-4-(6-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)哒嗪-3-基)-3-甲基哌嗪-1-羧酸盐(135mg,0.24mmol),收率:48%。
LC/MS(ESI+)calcd for C28H35ClN6O4(M+H+)m/z,555.2;found,555.2.
2.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((R)-2-甲基哌嗪-1-基)哒嗪-3-甲酰胺的合成
得到目标化合物,收率92%。LC/MS(ESI+)calcd for C23H27ClN6O2(M+H+)m/z455.1;found 455.1.
3.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((2R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-2-甲基哌嗪-1-基)哒嗪-3-甲酰胺的合成
得到目标化合物收率42%。LC/MS(ESI+)calcd for:C46H50ClFN9O6(M+H+)m/z,824.3;found 824.3.1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.64(d,J=8.2Hz,1H),7.84(dd,J=9.2,7.3Hz,2H),7.59(d,J=12.9Hz,1H),7.38(d,J=2.4Hz,1H),7.30(d,J=9.7Hz,1H),7.19-6.99(m,2H),5.07(dd,J=12.8,5.4Hz,1H),4.71-4.46(m,2H),4.28-4.16(m,1H),3.85(dt,J=9.6,4.2Hz,3H),3.60(d,J=9.5Hz,2H),3.22-3.04(m,2H),2.97-2.81(m,2H),2.42-2.23(m,3H),2.17-1.97(m,5H),1.89(d,J=12.3Hz,2H),1.63(q,J=5.5,3.2Hz,4H),1.56-1.44(m,2H),1.26-1.13(m,5H).
264:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((2R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基-2-甲基哌嗪-1-基)吡嗪-2-羧酰胺
1.叔丁基(R)-4-(5-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡嗪-2-基)-3-甲基哌嗪-1-羧酸盐的合成
得到目标化合物,收率52%。LC/MS(ESI+)calcd for C28H35ClN6O4(M+H+)m/z555.2;found 555.2.
2.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((R)-2-甲基哌嗪-1-基)吡嗪-2-甲酰胺的合成
得到目标化合物,收率92%。LC/MS(ESI+)calcdfor:C23H27ClN6O2(M+H+)m/z,455.1;found 455.1.
3.N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((2R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基-2-甲基哌嗪-1-基)吡嗪-2-羧酰胺的合成
得到目标化合物,收率42%。LC/MS(ESI+)calcdfor:C46H50ClFN9O6(M+H+)m/z,824.3;found,824.3.1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.61(d,J=1.2Hz,1H),8.20(s,1H),8.11(d,J=8.2Hz,1H),7.85(d,J=8.8Hz,1H),7.59(d,J=12.8Hz,1H),7.36(d,J=2.4Hz,1H),7.16-6.94(m,2H),5.07(dd,J=12.8,5.4Hz,1H),4.72-4.45(m,2H),4.21(d,J=12.7Hz,1H),3.84(dq,J=12.7,4.9,4.4Hz,3H),3.59(d,J=9.5Hz,2H),3.24-3.01(m,2H),2.94-2.79(m,2H),2.33(dd,J=11.1,6.9Hz,3H),2.13-1.96(m,5H),1.91-1.80(m,2H),1.70-1.43(m,6H),1.20(dd,J=17.2,6.6Hz,5H).
265:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)-3-甲基哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C46H50ClFN9O6(M+H+)m/z824.3;found 824.3.
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.62(d,J=8.2Hz,1H),7.84(dd,J=14.6,9.1Hz,2H),7.60(d,J=12.8Hz,1H),7.44-7.33(m,2H),7.19-7.05(m,2H),5.07(dd,J=12.8,5.4Hz,1H),4.54(dd,J=12.9,8.3Hz,1H),4.15(dd,J=39.2,12.6Hz,2H),3.84(dt,J=9.6,4.6Hz,3H),3.59(d,J=9.7Hz,2H),3.09-2.79(m,3H),2.43-2.27(m,2H),2.15-1.80(m,7H),1.70-1.57(m,4H),1.57-1.45(m,3H),1.23(s,2H),1.05(d,J=6.1Hz,3H),0.88-0.71(m,1H).
266:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-5-((3R)-4-((1R,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基-3-甲基哌嗪-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcdfor:C46H49ClFN9O6(M+H+)m/z824.3;found 824.3.
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.59(s,1H),8.26(s,1H),8.10(d,J=8.2Hz,1H),7.85(d,J=8.8Hz,1H),7.58(d,J=12.8Hz,1H),7.37(d,J=2.4Hz,1H),7.18-6.92(m,2H),5.06(dd,J=12.8,5.4Hz,1H),4.51(tt,J=9.9,4.3Hz,1H),4.13(t,J=12.5Hz,2H),3.83(dd,J=10.3,5.3Hz,3H),3.58(d,J=9.6Hz,2H),3.27(d,J=3.4Hz,1H),3.08-2.99(m,1H),2.97-2.80(m,2H),2.34(td,J=12.7,7.7Hz,2H),2.15-1.77(m,6H),1.71-1.41(m,7H),1.22(s,2H),1.05(d,J=6.1Hz,3H),0.91-0.70(m,1H).
267:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异丙吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-5-甲酰胺
LC/MS(ESI+)calcdfor:C44H48ClN9O6(M+H+)m/z,834.3;found,834.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.74(s,2H),8.11(d,J=7.4Hz,1H),7.86(d,J=8.7Hz,1H),7.63(d,J=8.2Hz,1H),7.38(d,J=2.4Hz,1H),7.14(dd,J=8.8,2.4Hz,1H),6.77(d,J=2.0Hz,1H),6.64(dd,J=8.4,2.1Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),4.72(d,J=12.8Hz,2H),4.59-4.48(m,1H),3.74(s,5H),3.01-2.81(m,3H),2.55(d,J=6.5Hz,1H),2.31(s,3H),2.11(t,J=6.6Hz,4H),2.03-1.95(m,2H),1.91(t,J=6.3Hz,2H),1.77(q,J=8.3,5.7Hz,6H),1.56-1.42(m,4H),1.24-1.13(m,2H),1.07-0.93(m,2H).
268:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)嘧啶-5-甲酰胺
LC/MS(ESI+)calcdfor:C44H47ClFN9O6(M+H+)m/z,852.3;found 852.3.
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.73(s,2H),8.11(d,J=7.5Hz,1H),7.88-7.84(m,1H),7.59(d,J=11.2Hz,1H),7.39(d,J=2.4Hz,1H),7.14(dd,J=8.8,2.5Hz,1H),6.89(d,J=7.6Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),4.71(d,J=12.8Hz,2H),4.54(dd,J=9.6,4.6Hz,1H),3.94-3.72(m,5H),2.99-2.79(m,3H),2.30(s,3H),2.10(q,J=7.2Hz,4H),2.04-1.97(m,1H),1.95-1.87(m,2H),1.77(q,J=5.5,4.9Hz,6H),1.57-1.41(m,4H),1.20(d,J=21.9Hz,3H),1.01(q,J=11.4,10.6Hz,2H),0.84(d,J=7.2Hz,1H).
269:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-7-氮杂螺环[3.5]壬-7-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.3;found,838.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.60(d,J=8.2Hz,1H),7.86(d,J=8.8Hz,1H),7.79(d,J=9.5Hz,1H),7.74(d,J=11.5Hz,1H),7.46(d,J=7.3Hz,1H),7.39(d,J=2.5Hz,1H),7.35(d,J=9.6Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.11(dd,J=12.8,5.4Hz,1H),4.52(d,J=10.4Hz,2H),4.03(d,J=7.1Hz,1H),3.85(s,2H),3.67(d,J=30.6Hz,5H),3.06(s,2H),2.88(m,4H),1.99(s,2H),1.89(d,J=12.4Hz,3H),1.63(d,J=9.6Hz,7H),1.58-1.45(m,7H).
270:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-7-氮杂螺[3.5]壬-2-基)哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H45ClFN9O6(M+H+)m/z838.3;found 838.3.
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.63(d,J=8.2Hz,1H),7.84(t,J=8.6Hz,2H),7.70(d,J=11.4Hz,1H),7.43(d,J=7.4Hz,1H),7.38(d,J=2.4Hz,1H),7.35(d,J=9.7Hz,1H),7.13(dd,J=8.8,2.5Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.53(dt,J=10.7,6.1Hz,1H),3.85(dd,J=7.8,3.8Hz,1H),3.70(d,J=5.2Hz,4H),2.96-2.83(m,1H),2.78-2.69(m,1H),2.65-2.55(m,1H),2.37(t,J=4.9Hz,4H),2.06(dt,J=32.9,6.7Hz,6H),1.94-1.83(m,2H),1.67(dt,J=27.9,5.5Hz,9H),1.52(t,J=11.0Hz,3H),1.22(s,2H).
271:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-7-氮杂螺环[3.5]壬-2-基)哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H46ClN9O6(M+H+)m/z820.3;found 820.3.
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.61(d,J=8.2Hz,1H),7.84(dd,J=9.2,7.8Hz,2H),7.64(d,J=8.5Hz,1H),7.40-7.29(m,3H),7.23(dd,J=8.8,2.3Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),4.52(dd,J=9.5,5.0Hz,1H),3.92-3.79(m,1H),3.70(s,4H),2.95-2.81(m,1H),2.73(dd,J=13.7,6.2Hz,1H),2.50(p,J=1.8Hz,6H),2.37(d,J=6.4Hz,4H),2.16-1.97(m,5H),1.89(d,J=11.4Hz,2H),1.64(d,J=9.7Hz,5H),1.56(s,4H),1.22(s,1H).
272:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(5-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌啶-4-基)甲基)六氢吡咯[3,4-c]吡咯-2(1H)-基)嘧啶-5-甲酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.3;found,838.3.1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.73(s,2H),8.15(d,J=7.4Hz,1H),7.86(d,J=8.8Hz,1H),7.70(d,J=11.4Hz,1H),7.47-7.30(m,2H),7.14(dd,J=8.8,2.5Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.60-4.47(m,1H),3.79(s,5H),3.64-3.54(m,3H),2.87(td,J=12.9,11.4,5.5Hz,3H),2.40-2.25(m,3H),2.22-2.06(m,4H),1.90(d,J=7.9Hz,2H),1.74(d,J=5.9Hz,5H),1.49(p,J=5.9,5.5Hz,5H),1.32-1.13(m,4H).
273:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(7-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮杂螺[3.5]壬-2-基)嘧啶-5-羧酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z838.3;found838.3.1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.74(s,2H),8.17(d,J=7.5Hz,1H),7.86(d,J=8.8Hz,1H),7.73(d,J=11.4Hz,1H),7.47(d,J=7.4Hz,1H),7.38(d,J=2.4Hz,1H),7.14(dd,J=8.8,2.5Hz,1H),5.11(dd,J=12.7,5.4Hz,1H),4.54(d,J=8.5Hz,1H),3.82(d,J=21.0Hz,5H),3.69(s,4H),3.10(dd,J=15.7,7.7Hz,3H),2.98-2.80(m,5H),2.14-2.05(m,3H),1.90(s,6H),1.70(s,2H),1.56-1.40(m,5H),1.18-1.12(m,1H).
274:N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2-,2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
LC/MS(ESI+)calcd for C44H47BrFN9O6(M+H+)m/z880.3;found 880.3.1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.2Hz,1H),7.81(t,J=9.4Hz,2H),7.63(d,J=8.3Hz,1H),7.49(d,J=2.4Hz,1H),7.33(d,J=9.7Hz,1H),7.16(dd,J=8.8,2.5Hz,1H),6.77(d,J=2.1Hz,1H),6.64(dd,J=8.4,2.1Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),4.58-4.41(m,3H),3.90-3.82(m,1H),3.75(s,4H),3.08-2.80(m,4H),2.64-2.53(m,2H),2.21(s,2H),2.14-2.06(m,2H),2.00(ddd,J=13.0,5.5,3.4Hz,1H),1.89(d,J=11.7Hz,4H),1.81(d,J=11.4Hz,6H),1.71-1.43(m,5H),1.22(s,1H),1.10(dd,J=18.1,6.7Hz,2H).
275:N-((1r,4r)-4-(4-氰基-3-甲基苯氧基)环己基)-6-(4-((2-(2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
LC/MS(ESI+)calcd for C45H51N9O6(M+H+)m/z814.4;found 814.4.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.3Hz,1H),7.80(d,J=9.5Hz,1H),7.64(dd,J=8.5,6.7Hz,2H),7.32(d,J=9.7Hz,1H),7.04(d,J=2.4Hz,1H),6.93(dd,J=8.6,2.5Hz,1H),6.77(d,J=2.1Hz,1H),6.64(dd,J=8.5,2.1Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),4.45(t,J=10.9Hz,3H),3.86(d,J=9.0Hz,1H),3.74(s,4H),2.99(t,J=12.4Hz,2H),2.89-2.81(m,1H),2.43(s,3H),2.32(s,3H),2.11(t,J=9.2Hz,3H),2.00(dd,J=8.9,4.3Hz,1H),1.91(s,6H),1.76(q,J=5.4Hz,5H),1.67-1.44(m,5H),1.22(s,1H),1.10(t,J=11.9Hz,2H).
276:N-((1r,4r)-4-(4-氰基-3-甲氧基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬基-7-基)甲基)哌啶-1-基)哒嗪-3-羧酰胺
LC/MS(ESI+)calcdfor C45H51N9O7(M+H+)m/z830.3;found 830.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.58(d,J=8.2Hz,1H),7.80(d,J=9.5Hz,1H),7.62(dd,J=10.5,8.7Hz,2H),7.33(d,J=9.7Hz,1H),6.78(d,J=2.1Hz,1H),6.72(dd,J=6.3,2.4Hz,2H),6.65(dd,J=8.4,2.1Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),4.49(dd,J=13.3,8.9Hz,3H),3.89(s,4H),3.75(s,4H),3.08-2.80(m,4H),2.12(d,J=12.9Hz,5H),2.06-1.96(m,2H),1.90(d,J=7.6Hz,4H),1.78(s,5H),1.67-1.42(m,5H),1.23(s,2H),1.18-1.02(m,2H).
277:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-7-氮杂螺[3.5]壬-2-基)哌嗪-1-基)吡嗪-2-甲酰胺
LC/MS(ESI+)calcdfor:C43H45ClFN9O6(M+H+)m/z,838.3;found,838.3.1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.60(d,J=1.2Hz,1H),8.25(s,1H),8.12(d,J=8.2Hz,1H),7.86(d,J=8.8Hz,1H),7.70(d,J=11.4Hz,1H),7.43(d,J=7.3Hz,1H),7.37(d,J=2.5Hz,1H),7.12(dd,J=8.8,2.5Hz,1H),5.10(dd,J=12.8,5.4Hz,1H),4.51(s,1H),3.82(d,J=10.1Hz,1H),3.69(s,4H),2.88(td,J=16.4,15.3,5.5Hz,1H),2.73(dd,J=13.2,5.6Hz,1H),2.36(d,J=5.9Hz,4H),2.15-1.98(m,5H),1.86(s,2H),1.75-1.44(m,11H),1.23(s,4H),0.84(d,J=7.0Hz,1H).
278:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-7-氮杂螺环[3.5]壬-2-基)哌嗪-1-基)吡嗪-2-甲酰胺LC/MS(ESI+)calcdfor:C43H46ClN9O6(M+H+)m/z,820.3;found,820.3.
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.60(s,1H),8.25(s,1H),8.11(d,J=8.1Hz,1H),7.85(d,J=8.7Hz,1H),7.64(d,J=8.4Hz,1H),7.33(dd,J=21.0,2.3Hz,2H),7.25-7.19(m,1H),7.11(dd,J=8.8,2.4Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),4.50(s,1H),3.92-3.78(m,1H),3.68(t,J=4.9Hz,5H),2.94-2.68(m,3H),2.34(t,J=5.0Hz,4H),2.04(dt,J=30.8,9.6Hz,6H),1.87(d,J=12.1Hz,3H),1.60(dd,J=23.8,11.0Hz,11H),1.22(s,1H).
279:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬烷-7-基)甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺
1.3-(甲硫基)-1,2,4-三嗪-6-羧酸甲酯
以3-氨基-6-溴-1,2,4-三嗪为原料,用文献方法(PCT Int.Appl.,2015182712)制备得到3-氨基-1,2,4-三嗪-6-羧酸甲酯。然后以其为原料用文献方法(PCT Int.Appl.,2015181539)制备得到淡黄色固体3-(甲硫基)-1,2,4-三嗪-6-羧酸甲酯。MS:calcd.forC6H7N3O2S[M+H]+:186.0;found:186.3.1H NMR(400MHz,CDCl3):δ8.95(s,1H),4.08(s,3H),2.73(s,3H).
2.甲基3-(4-(羟甲基)哌啶-1-基)-1,2,4-三嗪-6-羧酸酯
3-(甲硫基)-1,2,4-三嗪-6-羧酸甲酯(5.0g,27.00mmol),溶于200mL二氯甲烷中,分批加入间氯过氧苯甲酸(10.7g,62.09mmol)。室温搅拌4h。加入三乙胺(10.9g,107.99mmol),再加入4-哌啶甲醇(9.3g,80.99mmol)。室温搅拌过夜。加入水,和二氯甲烷,用碳酸钠调节pH至8-9,萃取,有机层用硫代硫酸钠洗涤,干燥,旋干,过硅胶柱纯化。得到化合物甲基3-(4-(羟甲基)哌啶-1-基)-1,2,4-三嗪-6-羧酸酯(2.0g,7.93mmol)。收率29.9%。LC/MS(ESI+)calcd for C11H17N4O3 +([M+H]+)m/z:253.1;found 253.1。
3.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-羟甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺的合成
甲基3-(4-(羟甲基)哌啶-1-基)-1,2,4-三嗪-6-羧酸酯(100.0mg,0.39mmol),4-(((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(149.1mg,0.59mmol)溶于2mL甲醇和0.3mL三乙胺加热回流36h。冷却至室温,旋干,硅胶柱层析纯化。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-羟甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺(72.0mg,0.15mmol)。收率38.5%。LC/MS(ESI+)calcd for C23H28ClN6O3 +([M+H]+)m/z:471.2;found471.1。
4.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-甲酰哌啶-1-基)-1,2,4-三嗪-6-甲酰胺的合成
N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-羟甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺(72.0mg,0.15mmol)溶于5mL二氯甲烷,加入Dess-Martin试剂(86mg,0.21mmol)。室温搅拌4h。加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,干燥,旋干。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-甲酰哌啶-1-基)-1,2,4-三嗪-6-甲酰胺(80mg,0.15mmol)。收率100.0%。
LC/MS(ESI+)calcd for C23H26ClN6O3 +([M+H]+)m/z:469.2;found 469.3。
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺的合成
N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-甲酰哌啶-1-基)-1,2,4-三嗪-6-甲酰胺(80mg,0.15mmol),2-(2,6-二氧哌啶-3-基)-5-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚啉-1,3-二酮(57.0mg,0.15mmol)溶于2mL二氯甲烷,加入1滴乙酸,最后加入三乙氧基硼氢化钠(126.0mg,0.60mmol)。室温搅拌过夜。加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,制备大板纯化。得到目标化合物(56mg,0.06mmol)。收率44.9%。
LC/MS(ESI+)calcd for C43H48ClN10O6([M+H]+)m/z:835.3;found 835.3。
1H NMR(400MHz,CDCl3)δ8.82(s,1H),8.15(s,1H),7.65(d,J=8.3Hz,1H),7.57(t,J=8.6Hz,2H),7.00(d,J=2.3Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),6.78(d,J=1.9Hz,1H),6.51(dd,J=8.3,1.9Hz,1H),4.94(dd,J=12.2,5.3Hz,3H),4.31(t,J=9.9Hz,1H),4.13–4.00(m,1H),3.75(s,4H),3.06(s,2H),2.93–2.66(m,4H),2.45(s,3H),2.35–2.07(m,8H),1.96(s,8H),1.72–1.63(m,2H),1.44(dd,J=17.2,8.6Hz,2H).
280:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)哌啶-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C43H47ClN9O6 +([M+H]+)m/z:820.3;found 820.3。
1H NMR(400MHz,CDCl3)δ8.84(d,J=0.9Hz,1H),8.20(s,1H),7.99(s,1H),7.65(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.40(d,J=8.3Hz,1H),6.99(d,J=2.3Hz,1H),6.85(dd,J=8.7,2.3Hz,1H),6.78(d,J=1.9Hz,1H),6.51(dd,J=8.3,1.9Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.54(d,J=12.5Hz,2H),4.30(t,J=10.0Hz,1H),4.10–3.96(m,1H),3.75(s,4H),3.00(t,J=11.9Hz,2H),2.91–2.67(m,5H),2.17(dd,J=9.1,5.0Hz,5H),2.01(d,J=31.7Hz,5H),1.75–1.64(m,8H),1.52–1.44(m,2H).
281:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-(2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H47ClN9O6 +([M+H]+)m/z:820.3;found 820.3。
1H NMR(400MHz,CDCl3)δ8.14(s,1H),8.01(d,J=9.4Hz,1H),7.88(d,J=8.2Hz,1H),7.65(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.01(t,J=6.0Hz,2H),6.86(dd,J=8.7,2.3Hz,1H),6.78(d,J=1.7Hz,1H),6.52(d,J=8.4Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.61(s,2H),4.32(s,1H),4.06(d,J=8.0Hz,1H),3.76(s,4H),3.07(t,J=12.5Hz,2H),2.94–2.83(m,2H),2.82–2.67(m,3H),2.28–1.87(m,10H),1.64(s,8H),1.50–1.42(m,2H).
282:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C44H49ClN9O6 +([M+H]+)m/z:834.3;found 834.3。
1H NMR(400MHz,CDCl3)δ8.23(s,1H),7.98(d,J=9.2Hz,1H),7.88(d,J=8.2Hz,1H),7.68(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.28(d,J=2.2Hz,1H),7.05(dd,J=8.6,2.3Hz,1H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),6.59(d,J=9.3Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.38-4.26(m,1H),4.15-3.79(m,7H),2.98(t,J=12.1Hz,2H),2.92-2.71(m,3H),2.42(s,4H),2.15(dt,J=12.8,7.9Hz,6H),1.92(s,5H),1.71(dd,J=19.8,10.2Hz,8H),1.46(dt,J=14.3,7.3Hz,2H).
283:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)吡嗪-3-甲酰胺
LC/MS(ESI+)calcd for C44H49ClN9O6 +([M+H]+)m/z:834.3;found 834.3。
1H NMR(400MHz,CDCl3)δ8.84(s,1H),8.01(d,J=8.8Hz,1H),7.70(d,J=8.2Hz,1H),7.62(d,J=8.5Hz,1H),7.55(d,J=8.7Hz,1H),7.39(d,J=2.2Hz,1H),7.30(dd,J=8.6,2.3Hz,1H),7.06(d,J=2.4Hz,1H),6.99(dd,J=8.8,2.4Hz,1H),6.86(d,J=9.3Hz,1H),4.99–4.92(m,1H),4.35–4.28(m,1H),3.98(m,6H),3.69–3.61(m,1H),2.95(d,J=55.1Hz,9H),2.18(m,6H),2.00(m,5H),1.55–1.43(m,10H).
284:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(7-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺[3.5]壬-2-基)吡嗪-2-甲酰胺
LC/MS(ESI+)calcd for C43H46ClFN9O6 +([M+H]+)m/z:838.3;found 838.2。
1H NMR(400MHz,CDCl3)δ8.12(s,1H),7.99(d,J=9.6Hz,1H),7.87(d,J=8.0Hz,1H),7.56(d,J=8.7Hz,1H),7.36(d,J=10.9Hz,1H),7.04–6.96(m,2H),6.86(dd,J=8.8,2.1Hz,1H),6.80(d,J=7.5Hz,1H),4.91(dd,J=12.2,5.0Hz,1H),4.57(d,J=13.0Hz,2H),4.32(s,1H),4.05(s,1H),3.89(s,4H),3.79(s,1H),3.46(d,J=6.7Hz,1H),3.20(s,1H),3.05(t,J=12.3Hz,2H),2.89(d,J=14.4Hz,1H),2.82–2.68(m,3H),2.17(d,J=10.9Hz,5H),2.00(s,4H),1.66(dd,J=24.2,11.3Hz,8H),1.49(d,J=12.1Hz,2H).
285:N-((1r,4r)-4-(4-氰基-3-(三氟甲基)苯氧基)环己基)-6-(2-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)-2,7-二氮杂螺环[3.5]壬-7-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcdfor C444H49ClN9O6 +([M+H]+)m/z:886.4;found 886.3。
1H NMR(400MHz,CDCl3)δ8.39(s,1H),7.99(d,J=9.5Hz,1H),7.88(d,J=8.1Hz,1H),7.75(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.38(d,J=7.3Hz,1H),7.25(s,1H),7.10(dd,J=8.7,2.3Hz,1H),7.00(d,J=9.6Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.39(t,J=10.1Hz,1H),4.07(d,J=8.2Hz,1H),3.72(s,4H),3.64(d,J=12.3Hz,2H),3.28(s,2H),2.96–2.72(m,5H),2.58(s,2H),2.27–2.09(m,5H),1.90(d,J=14.8Hz,5H),1.79–1.59(m,8H),1.46(s,2H).
286:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡啶酰胺
1.4-(6-(甲氧羰基)吡啶-3-基)哌嗪-1-羧酸叔丁酯的合成
氮气保护下,5-溴海松酸甲酯(500.0mg,2.31mmol),Boc-哌嗪(646.0mg,3.47mmol),Pd2(dba)3(100.0mg,0.23mmol),BINAP(100.0mg,0.23mmol),Cs2CO3(1.51g,4.62mmol)加入到20mL甲苯中。加热到100℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物4-(6-(甲氧羰基)吡啶-3-基)哌嗪-1-羧酸叔丁酯(450.0mg,1.40mmol)。收率60.5%。LC/MS(ESI+)calcd for C16H24N3O4 +([M+H]+)m/z:322.2;found 322.0。
2.化合物5-(4-(叔丁氧羰基)哌嗪-1-基)吡啶酸的合成
4-(6-(甲氧羰基)吡啶-3-基)哌嗪-1-羧酸叔丁酯(450.0mg,1.40mmol),溶于到5mL甲醇,加入5mL 2N NaOH溶液。室温搅拌4h。用0.5N HCl调节pH至4-5,加入乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干。得到化合物5-(4-(叔丁氧羰基)哌嗪-1-基)吡啶酸(450.0mg,1.40mmol)。收率100.0%。
3.化合物4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡啶-3-基)哌嗪-1-羧酸叔丁酯的合成
5-(4-(叔丁氧羰基)哌嗪-1-基)吡啶酸(200.0mg,0.65mmol),HATU(260.0mg,0.71mmol),DIEA(252.0mg,1.95mmol)加入到5mL二氯甲烷中,再加入4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(163.0mg,0.65mmol)。室温搅拌2h。加水和乙酸乙酯,萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡啶-3-基)哌嗪-1-羧酸叔丁酯(300.0mg,0.55mmol)。收率85.5%。
4.N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(哌嗪-1-基)吡啶酰胺的合成
氮4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰)吡啶-3-基)哌嗪-1-羧酸叔丁酯(300mg,0.55mmol)溶于5mL二氯甲烷和5mL三氟乙酸中。室温搅拌2h。浓缩,旋干。得到化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(哌嗪-1-基)吡啶酰胺(250mg,0.55mmol)。收率100.0%。
5.化合物N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((1r,5S,6s)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-基)甲基)哌嗪-1-基)吡啶酰胺的合成
N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(哌嗪-1-基)吡啶酰胺(50.0mg,0.11mmol),(1R,5S)-3-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-3-氮杂双环[3.1.0]正己烷-6-甲醛(43.8mg,0.11mmol)溶于2mL二氯甲烷,加入1滴乙酸,最后加入三乙氧基硼氢化钠(72.0mg,0.33mmol)。室温搅拌过夜。加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,制备大板纯化。得到目标化合物(50mg,0.06mmol)。收率54.3%。LC/MS(ESI+)calcd for C42H43ClFN8O6 +([M+H]+)m/z:809.3;found 809.2。
1H NMR(400MHz,CDCl3)δ8.51(s,1H),8.18(d,J=2.7Hz,1H),8.06(d,J=8.7Hz,1H),7.74(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=12.5Hz,1H),7.25–7.19(m,1H),7.05(d,J=7.4Hz,1H),7.00(d,J=2.3Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),4.92(dd,J=12.2,5.3Hz,1H),4.31(dd,J=12.0,8.3Hz,1H),4.02(dd,J=11.3,7.3Hz,1H),3.90(d,J=10.1Hz,2H),3.60(d,J=9.3Hz,2H),3.43(s,4H),2.96–2.68(m,6H),2.52(s,2H),2.15(dd,J=18.8,7.6Hz,5H),1.69(d,J=12.0Hz,8H),1.53–1.41(m,2H).
287:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)哌啶-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C43H46ClFN9O6 +([M+H]+)m/z:838.3;found 838.3。
1H NMR(400MHz,CDCl3)δ8.83(s,1H),8.22(s,1H),7.97(s,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=10.8Hz,1H),7.43–7.35(m,2H),7.01(t,J=6.6Hz,1H),6.85(dd,J=8.7,2.3Hz,1H),4.93(dd,J=12.3,5.2Hz,1H),4.43(s,2H),4.31(t,J=10.1Hz,1H),4.03(d,J=8.1Hz,1H),3.50(s,2H),3.14(d,J=31.2Hz,6H),2.94––2.71(m,4H),2.17(d,J=8.4Hz,5H),1.95(s,4H),1.66(d,J=9.9Hz,8H),1.47(d,J=11.8Hz,2H).
288:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-7-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C43H47ClN9O6 +([M+H]+)m/z:820.3;found 820.3。
1H NMR(400MHz,CDCl3)δ8.83(d,J=1.2Hz,1H),8.17(s,1H),7.97(d,J=1.2Hz,1H),7.67(d,J=8.5Hz,1H),7.56(d,J=8.7Hz,1H),7.39(d,J=8.2Hz,1H),7.28(s,1H),7.05(dd,J=8.6,2.3Hz,1H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.30(d,J=3.7Hz,1H),4.03(d,J=8.1Hz,1H),3.87(d,J=13.1Hz,2H),3.76–3.53(m,4H),3.22(s,4H),3.08(t,J=10.5Hz,2H),2.94–2.70(m,3H),2.54–2.42(m,1H),2.22–2.13(m,5H),1.89(s,6H),1.69(dd,J=22.2,9.7Hz,8H),1.49–1.41(m,2H).
289:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)-2,7-二氮螺环[3.5]壬-7-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C43H46ClFN9O6 +([M+H]+)m/z:838.3;found 838.3。
1H NMR(400MHz,CDCl3)δ8.83(d,J=1.2Hz,1H),8.33(s,1H),7.98(d,J=1.1Hz,1H),7.56(d,J=8.7Hz,1H),7.46(d,J=11.0Hz,1H),7.39(dd,J=7.7,4.1Hz,2H),6.99(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.30(dd,J=11.9,8.2Hz,1H),4.02(dd,J=11.6,7.4Hz,1H),3.73–3.57(m,6H),3.24(s,4H),2.93(t,J=10.5Hz,2H),2.84–2.72(m,2H),2.44(s,1H),2.20–2.13(m,4H),1.88(d,J=12.6Hz,10H),1.74–1.55(m,6H),1.47(dd,J=22.6,10.3Hz,2H).
290:(3aR,5s,6aS)-2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基4-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)哒嗪-3-基)哌嗪-1-羧酸酯
LC/MS(ESI+)calcd for C43H46ClFN9O6 +([M+H]+)m/z:868.3;found867.7。
1H NMR(400MHz,CDCl3)δ8.24(s,1H),8.07(d,J=9.4Hz,1H),7.91(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=12.1Hz,1H),7.09(d,J=7.4Hz,1H),7.04(d,J=9.5Hz,1H),7.00(d,J=2.3Hz,1H),6.86(dd,J=8.7,2.4Hz,1H),5.33(d,J=2.6Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.31(dd,J=11.8,8.2Hz,1H),4.06(d,J=8.0Hz,1H),3.81(s,4H),3.60(d,J=7.5Hz,2H),3.45(d,J=10.5Hz,2H),2.98(s,2H),2.93–2.72(m,3H),2.39–2.02(m,8H),1.99–1.81(m,5H),1.69(dd,J=22.2,9.9Hz,2H),1.55–1.43(m,2H).
291:N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-6-(4-((1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉)哌啶-4-基)氨基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C43H48ClFN9O6 +([M+H]+)m/z:840.3;found 840.3。
1H NMR(400MHz,CDCl3)δ8.15(d,J=9.2Hz,1H),8.00(d,J=9.5Hz,1H),7.57(d,J=8.7Hz,1H),7.47(d,J=10.9Hz,1H),7.39(d,J=7.2Hz,1H),7.03(d,J=9.6Hz,1H),6.97(d,J=2.3Hz,1H),6.81(dd,J=8.7,2.3Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.51(d,J=12.2Hz,2H),4.19(d,J=9.1Hz,1H),4.07(s,1H),3.79–3.62(m,2H),3.17(t,J=11.7Hz,3H),3.00–2.88(m,3H),2.85–2.62(m,3H),2.12(s,5H),1.74(d,J=21.4Hz,5H),1.28(s,6H),1.21(s,6H).
292:N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺
1.化合物N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,6aS)-5-氧代六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺的合成
化合物6-氯-N-((1s,4s)-4-(3-氯-4-氰基苯氧基)环己基)哒嗪-3-甲酰胺(200.0mg,0.5mmol),(3aR,6aS)-六氢环戊[c]吡咯-5(1H)-酮(128.0mg,1.0mmol)溶于5mLDMF,加入碳酸钾(141.0mg,1.0mmol)。加热到80℃搅拌过夜。加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,硅胶柱层析纯化。得到化合物N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,6aS)-5-氧代六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(70.0mg,0.14mmol)。收率28.5%。
2.N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺的合成
化合物N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,6aS)-5-氧代六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(60.0mg,0.12mmol),2-(2,6-二氧哌啶-3-基)-5-氟-6-(哌嗪-1-基)异苄-1,3-二酮(45.0mg,0.12mmol)溶于2mL二氯甲烷和5mL甲醇,加入1滴乙酸,最后加入氰基硼氢化钠(31.0mg,0.50mmol)。室温搅拌过夜。加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,制备大板纯化。得到化合物N-((1s,4S)-4-(3-氯-4-氰基苯氧基)环己基)-6-((3aR,5R,6aS)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-基)哒嗪-3-甲酰胺(30.0mg,0.04mmol)。收率29.1%。
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.2。
1H NMR(400MHz,CDCl3)δ8.41(s,1H),7.98(d,J=9.4Hz,1H),7.90(d,J=8.2Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=10.9Hz,1H),7.41(d,J=7.2Hz,1H),7.01(d,J=2.4Hz,1H),6.86(dd,J=8.8,2.4Hz,1H),6.72(d,J=9.4Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.38–4.28(m,1H),4.05(dd,J=11.4,7.3Hz,1H),3.76(d,J=6.5Hz,2H),3.66(s,2H),3.49–3.18(m,4H),2.90(d,J=10.2Hz,2H),2.76(dq,J=13.4,8.9Hz,4H),2.42–2.26(m,2H),2.16(ddd,J=12.0,10.1,3.1Hz,5H),1.85–1.61(m,8H),1.46(dd,J=12.4,9.8Hz,2H).
293:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-((3aR,5R,6aS)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.3。
1H NMR(400MHz,CDCl3)δ8.84(d,J=1.3Hz,1H),8.38(s,1H),7.72(d,J=1.2Hz,1H),7.56(d,J=8.7Hz,1H),7.48(d,J=10.9Hz,1H),7.40(d,J=7.5Hz,2H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.38–4.24(m,1H),4.03(dd,J=7.4,3.4Hz,1H),3.74(dd,J=11.0,7.5Hz,2H),3.61(d,J=10.3Hz,2H),3.33(s,4H),2.88–2.73(m,6H),2.43–2.25(m,2H),2.15(dt,J=7.5,6.4Hz,5H),1.81–1.58(m,8H),1.48(dd,J=16.8,6.3Hz,2H).
294:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-((3aR,5R,6aS)-5-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-基)嘧啶-2-羧酰胺
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.2。
295:N-((1s,4s)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢旋环戊[c]吡咯-5-基)哌嗪-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.2。
295中间体的合成:
1.化合物(3aR,5r,6aS)-叔丁基5-(4-((苄氧基)羰基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-羧酸酯的合成
化合物(3aR,6aS)-叔丁基5-氧六氢环戊烷[c]吡咯-2(1H)-羧酸酯(1.0g,4.4mmol),苄基哌嗪-1-羧酸酯(1.0g,4.4mmol)溶于20mL二氯甲烷,加入0.2mL乙酸,再加入氰基硼氢化钠(836.0mg,13.2mmol)。室温搅拌过夜。加入水和二氯甲烷萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(3aR,5r,6aS)-叔丁基5-(4-((苄氧基)羰基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-羧酸(780.0mg,1.8mmol)。收率40.9%。
2.化合物(3aR,5r,6aS)-叔丁基5-(哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-羧酸酯
化合物(3aR,5r,6aS)-叔丁基5-(4-((苄氧基)羰基)哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-羧酸(780.0mg,1.8mmol)甲醇20mL,加入湿Pd/C(300.0mg),用氢气球置换3次。室温搅拌过夜。垫硅藻土抽滤,用甲醇洗涤2次。滤液旋干,得到粗品化合物(3aR,5r,6aS)-叔丁基5-(哌嗪-1-基)六氢环戊[c]吡咯-2(1H)-羧酸酯(530.0mg,1.8mmol)。收率98.8%。
296:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)八氢环戊[c]吡咯-5-基)哌嗪-1-基)吡嗪-2-羧酰胺
LC/MS(ESI+)calcd for C42H45ClN9O6 +([M+H]+)m/z:806.3;found 806.2。
1H NMR(400MHz,CDCl3)δ8.86(d,J=1.1Hz,1H),8.23(s,1H),7.97(s,1H),7.66(d,J=8.4Hz,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=8.3Hz,1H),6.98(dd,J=10.2,2.2Hz,2H),6.84(dd,J=8.8,2.4Hz,1H),6.71(dd,J=8.5,2.1Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.30(t,J=10.2Hz,1H),4.04(d,J=4.0Hz,1H),3.93(d,J=51.6Hz,4H),3.68–3.58(m,2H),3.48(s,2H),2.91–2.72(m,7H),2.33(d,J=5.9Hz,2H),2.23–2.06(m,5H),1.69(d,J=12.1Hz,8H),1.52–1.43(m,2H).
297:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.2。
1H NMR(400MHz,CDCl3)δ8.28(s,1H),8.02(d,J=9.5Hz,1H),7.89(d,J=8.3Hz,1H),7.56(d,J=8.7Hz,1H),7.41(d,J=12.2Hz,1H),7.10(d,J=7.4Hz,1H),6.99(t,J=6.3Hz,2H),6.85(dd,J=8.8,2.4Hz,1H),4.92(dd,J=12.2,5.3Hz,1H),4.36–4.25(m,1H),4.05(dd,J=11.5,7.3Hz,1H),3.84(s,4H),3.59(s,4H),2.95–2.75(m,6H),2.39–2.26(m,2H),2.25–2.09(m,5H),1.68(dd,J=22.1,9.7Hz,8H),1.50–1.42(m,2H).
298:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)哌嗪-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)calcd for C42H45ClN9O6 +([M+H]+)m/z:806.3;found 806.2。
1H NMR(400MHz,CDCl3)δ8.30(s,1H),8.02(d,J=9.5Hz,1H),7.89(d,J=8.2Hz,1H),7.66(d,J=8.4Hz,1H),7.56(d,J=8.7Hz,1H),6.99(dd,J=10.2,2.2Hz,3H),6.85(dd,J=8.7,2.4Hz,1H),6.70(dd,J=8.5,2.1Hz,1H),4.94(dd,J=12.3,5.3Hz,1H),4.31(t,J=3.7Hz,1H),4.13–4.00(m,1H),3.83(s,4H),3.65–3.55(m,2H),3.43(d,J=9.1Hz,2H),2.89–2.73(m,6H),2.38–2.26(m,2H),2.15(dt,J=12.7,7.3Hz,5H),1.68(dd,J=22.2,9.8Hz,8H),1.50–1.42(m,2H).
299:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)哌嗪-1-基)嘧啶-5-甲酰胺
LC/MS(ESI+)calcd for C42H44ClFN9O6 +([M+H]+)m/z:824.3;found 824.2。
1H NMR(400MHz,CDCl3)δ8.69(s,2H),8.36(s,1H),7.56(d,J=8.6Hz,1H),7.40(d,J=12.1Hz,1H),7.09(d,J=7.1Hz,1H),6.99(s,1H),6.85(d,J=8.4Hz,1H),5.89(s,1H),4.92(d,J=6.6Hz,1H),4.29(s,1H),3.97(s,4H),3.58(s,4H),2.82(dd,J=58.5,14.3Hz,6H),2.30(s,2H),2.17(d,J=11.6Hz,5H),1.84–1.50(m,8H),1.43(d,J=10.9Hz,2H).
300:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(4-((3aR,5r,6aS)-2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基)哌嗪-1-基)嘧啶-5-羧酰胺
LC/MS(ESI+)calcd for C42H45ClN9O6 +([M+H]+)m/z:806.3;found 806.3.
1H NMR(400MHz,CDCl3)δ8.68(s,2H),8.36(s,1H),7.65(d,J=8.4Hz,1H),7.56(d,J=8.7Hz,1H),6.98(dd,J=7.8,2.2Hz,2H),6.84(dd,J=8.8,2.4Hz,1H),6.69(dd,J=8.6,2.1Hz,1H),5.85(d,J=7.7Hz,1H),4.94(dd,J=12.3,5.4Hz,1H),4.35–4.22(m,1H),4.12–3.80(m,5H),3.67–3.51(m,2H),3.47–3.35(m,2H),2.93–2.70(m,6H),2.35–2.24(m,2H),2.24–2.09(m,5H),1.79–1.56(m,8H),1.42(dd,J=14.9,8.8Hz,2H).
301:(3aR,5s,6aS)-2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)八氢环戊[c]吡咯-5-基4-(5-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨甲酰基)吡嗪-2-基)哌嗪-1-羧酸酯
LC/MS(ESI+)calcd for C43H45ClN9O8 +([M+H]+)m/z:850.3;found 850.2。
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.63(d,J=1.2Hz,1H),8.27(s,1H),8.15(d,J=8.4Hz,1H),7.86(d,J=8.8Hz,1H),7.65(d,J=8.4Hz,1H),7.37(d,J=2.4Hz,1H),7.12(dd,J=8.8,2.4Hz,1H),6.94(s,1H),6.90–6.80(m,1H),5.16(s,1H),5.06(dd,J=12.8,5.4Hz,1H),4.51(t,J=9.8Hz,1H),3.83(d,J=8.0Hz,1H),3.76–3.68(m,4H),3.59(dd,J=10.2,7.4Hz,2H),3.51(s,4H),3.30(s,2H),2.98(s,2H),2.93–2.83(m,1H),2.63–2.52(m,2H),2.09(d,J=10.5Hz,2H),2.00(dd,J=9.1,3.9Hz,3H),1.89(dd,J=14.7,8.0Hz,4H),1.66–1.46(m,4H).
302:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氨基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcd forC43H47ClN7O6 +([M+H]+)m/z:792.3;found 792.2.
1H NMR(400MHz,CDCl3)δ7.67(dd,J=13.0,8.7Hz,3H),7.56(d,J=8.8Hz,1H),7.35(s,1H),7.18–7.11(m,1H),7.08(s,1H),6.99(s,1H),6.90(d,J=9.2Hz,2H),6.85(d,J=8.6Hz,1H),4.98–4.91(m,1H),4.33–4.27(m,1H),4.10–3.90(m,4H),3.87–3.76(m,2H),3.09–3.00(m,2H),2.89(s,3H),2.78–2.73(m,1H),2.17(m,6H),2.04(m,6H),1.60–1.49(m,7H).
303:N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊烯-2-基)-5-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)哌嗪-2-羧酰胺
1.化合物(3a'R,6a'S)-5,5-二甲基四氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'(3'H)-1
(3as,6as)-四氢戊烯-2,5(1H,3H)-二酮(6.0g,43.4mmol),2,2-二甲基丙烷-1,3-二醇(4.5g,43.4mmol),加入到200mL中,再加入对甲苯磺酸(150.0mg,43.4mmol)。加热回流4h。冷却至室温。浓缩,旋干,过硅胶柱纯化。得到化合物(3a'R,6a'S)-5,5-二甲基四氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'(3'H)-1(4.5g,20.0mmol)。收率46%。
2.化合物(3a'R,5's,6a's)-5,5-二甲基六氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'-醇
(3a'R,6a'S)-5,5-二甲基四氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'(3'H)-1(4.5g,20.0mmol)溶于甲醇30mL和二氯甲烷30mL,分批加入硼氢化钠(1.5g,40.0mmol)。室温搅拌2h。加入水和二氯化甲烷,萃取,有机层用饱和食盐水洗涤,干燥,旋干。得到粗品化合物(3a'R,5's,6a's)-5,5-二甲基六氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'-醇(3.5g,15.5mmol)。收率77.1%。
3.化合物(3aR,5s,6aS)-5-羟基六氢戊烯-2(1H)-酮
(3a'R,5's,6a's)-5,5-二甲基六氢-1'H-螺环[[1,3]二氧六环-2,2'-戊烯]-5'-醇(2.5g,11.5mmol)溶于丙酮50mL,加入4-甲基苯磺酸吡啶(1.1g,4.6mmol)。加热回流4h。冷却至室温,加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(3aR,5s,6aS)-5-羟基六氢戊烯-2(1H)-酮(0.9g,6.2mmol)。收率58.1%。
4.化合物(2r,3aR,5s,6aS)-5-(二苄基氨基)八氢戊烯-2-醇
化合物(3aR,5s,6aS)-5-羟基六氢戊烯-2(1H)-酮(0.6g,4.3mmol),二苄胺(0.8g,4.3mmol)溶于乙腈20mL,加入乙酸(0.2g,4.3mmol),再加入氰基硼氢化钠(0.8g,12.8mmol)。室温搅拌过夜。加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(2r,3aR,5s,6aS)-5-(二苄基氨基)八氢戊烯-2-醇(250.0mg,6.2mmol)。收率18.1%。
5.化合物(2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-醇
化合物(2r,3aR,5s,6aS)-5-(二苄基氨基)八氢戊烯-2-醇(250.0mg,0.7mmol)溶于THF 5mL,加入湿Pd/C(100.0mg),用氢气球置换3次。室温搅拌过夜。垫硅藻土抽滤,用甲醇洗涤2次。滤液旋干,得到粗品化合物(2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-醇(35.0mg,0.2mmol)。收率32.8%。
6.化合物4-((2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-基)氧基)-2-氯苯甲腈
化合物(2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-醇(35.0mg,0.2mmol)溶于DMF 3mL,冰浴,加入钠氢(30.0mg,0.6mmol)。冰浴下,搅拌1h。加入2-氯-4-氟苯甲腈(30.0mg,0.2mmol)。室温搅拌1h。加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物4-((2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-基)氧基)-2-氯苯甲腈(25mg,0.09mmol)。收率36.4%。
7.化合物5-氯-N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊-2-基)吡嗪-2-甲酰胺
5-氯吡嗪-2-羧酸(14.0mg,0.09mmol),化合物4-((2r,3aR,5s,6aS)-5-氨基八氢戊烯-2-基)氧基)-2-氯苯甲腈(25.0mg,0.09mmol),HATU(32.0mg,0.09mmol)加入到DMF1mL,再加入DIEA(23.0mg,0.18mmol)。室温搅拌1h。加入二氯甲烷和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物5-氯-N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊-2-基)吡嗪-2-甲酰胺(30.0mg,0.07mmol)。收率80.0%。
8.化合物叔丁基4-((1-(5-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊基-2-氨甲酰)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯
5-氯-N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊-2-基)吡嗪-2-甲酰胺(30.0mg,0.07mmol),4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(40.1mg,0.14mmol),DIEA(27.8mg,0.21mmol)加入到二氧六环2mL,加热到100℃,搅拌6h。冷却至室温。加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物叔丁基4-((1-(5-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊基-2-氨甲酰)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯(30.0mg,0.45mmol)。收率62.8%。
9.化合物N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊烯-2-基)-5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-甲酰胺
化合物叔丁基4-((1-(5-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊基-2-氨甲酰)吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸酯(30.0mg,0.45mmol)溶于三氟乙酸1mL和二氯甲烷1mL,室温搅拌1h。旋干得到粗品化合物N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊烯-2-基)-5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-甲酰胺(25.0mg,0.45mmol)。收率100.0%。
10.化合物N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊烯-2-基)-5-(4-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1
化合物N-((2s,3aR,5r,6aS)-5-(3-氯-4-氰基苯氧基)八氢戊烯-2-基)-5-(4-(哌嗪-1-基甲基)哌啶-1-基)吡嗪-2-甲酰胺(25.0mg,0.45mmol),2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(12.0mg,0.45mmol),DIEA(57.0mg,4.5mmol)加入到1mLDMSO中。加热到130℃搅拌2h。冷却至室温,加入乙酸乙酯和水萃取,有机层用饱和食盐水洗涤,干燥,旋干,制备大板纯化。得到目标化合物(14mg,0.17mmol)。收率38.8%。
LC/MS(ESI+)calcd for C43H47ClN9O6 +([M+H]+)m/z:820.3;found 820.3.
1H NMR(400MHz,CDCl3)δ8.82(d,J=1.2Hz,1H),8.09(s,1H),7.97(s,1H),7.72(d,J=8.5Hz,1H),7.57(d,J=8.7Hz,1H),7.47(d,J=8.3Hz,1H),7.30(d,J=1.9Hz,1H),7.08(d,J=8.4Hz,1H),7.03(d,J=2.4Hz,1H),6.86(dd,J=8.7,2.4Hz,1H),4.95(dd,J=12.3,5.2Hz,1H),4.86(s,1H),4.48(d,J=13.1Hz,2H),4.33(d,J=8.0Hz,1H),3.49(s,2H),3.00(t,J=12.3Hz,2H),2.90(d,J=16.9Hz,1H),2.85(s,1H),2.83–2.72(m,2H),2.67(d,J=24.9Hz,4H),2.47–2.33(m,3H),2.17(td,J=14.9,7.3Hz,3H),2.01(dd,J=14.9,8.1Hz,2H),1.89(d,J=13.9Hz,2H),1.63(s,7H),1.48(dd,J=17.5,8.9Hz,2H).
304:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)氧基)哌啶-1-基)苯甲酰胺
LC/MS(ESI+)calcd forC43H46ClN9O6 +([M+H]+)m/z:793.3;found 793.2.
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),7.97(d,J=7.6Hz,1H),7.86(d,J=8.8Hz,1H),7.74(d,J=8.8Hz,2H),7.66(d,J=8.5Hz,1H),7.39(d,J=2.3Hz,1H),7.34(s,1H),7.26(d,J=8.6Hz,1H),7.15(dd,J=8.8,2.3Hz,1H),6.96(t,J=10.5Hz,2H),5.08(dd,J=12.9,5.4Hz,1H),4.53(s,1H),3.90-3.74(m,4H),3.74–3.57(m,3H),3.26(t,J=10.0Hz,2H),3.03(t,J=9.9Hz,2H),2.94–2.81(m,1H),2.64–2.55(m,2H),2.11(d,J=9.0Hz,2H),2.06–1.99(m,1H),1.90(d,J=9.5Hz,6H),1.54(dd,J=21.9,11.9Hz,8H).
305:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基烟酰胺
LC/MS(ESI+)calcd forC43H46D3ClFN8O5 +m/z 814.4;found 814.3
306:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-3-氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-N-氘代甲基烟酰胺
LC/MS(ESI+)calcd forC43H46D3ClFN8O5 +m/z 814.4;found 814.3
307:2-氯-4-((1r,4r)-4-(2-(4-((4-(6-氟-2-(1-甲基-2,6-二氧哌啶-3-基)-3-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈
LC/MS(ESI+)calcd for C44H49ClFN8O5 +m/z 823.3;found 823.3.
308:(3-(5-(4-((1-(6-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)哌啶-4-基)甲基)哌嗪-1-基)-6-氟-1-氧异吲哚-2-基)-2,6-二氧哌啶-1-基)甲基2,5,8,11-四氧哌啶-13-基酰胺
LC/MS(ESI+)calcd for C54H67ClFN8O12+m/z 1073.5;found 1073.4.
309:2-氯-4-((1r,4r)-4-(2-(4-((4-(3-(2,6-二氧哌啶-3-基)-2-甲基-4-氧代-3,4-二氢喹唑啉-7-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己基)氧基)苯甲腈
LC/MS(ESI+)calcd for C44H49ClN9O5 +m/z 818.4;found 818.4.
1H NMR(400MHz,CDCl3)δ8.20(s,1H),8.03(d,J=8.9Hz,1H),7.83(d,J=8.8Hz,1H),7.58(d,J=8.7Hz,1H),7.03(t,J=4.4Hz,2H),6.95(s,1H),6.88(dd,J=8.7,2.3Hz,1H),6.68(d,J=8.9Hz,1H),4.74(dd,J=11.2,5.9Hz,1H),4.50(d,J=12.4Hz,2H),4.31(s,2H),4.21(s,2H),3.55(d,J=31.2Hz,4H),2.97(dd,J=26.9,15.2Hz,4H),2.84–2.71(m,2H),2.68(s,3H),2.26(s,2H),2.19(d,J=5.5Hz,1H),2.03(s,4H),1.84–1.64(m,10H).
310:2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂环丁烷-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物(2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚-2-基)环丁氧基)苯甲腈的合成
氮气保护下,4-((1r,3r)-3-(5-溴-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(500.0mg,1.06mmol),乙炔基三甲基硅烷(518.0mg,5.28mmol),Pd(PPh3)2Cl2(100.0mg,0.1mmol),CuI(60.0mg,0.2mmol),2mL三乙胺加入到6mL甲苯中。加热到100℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚-2-基)环丁氧基)苯甲腈(400mg,0.80mmol)。收率77.2%。
LC/MS(ESI+)calcd for C28H32ClN2O2Si+([M+H]+)m/z:491.2;found 491.1。
2.化合物2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(1-氧-5-((三甲硅基)乙炔基)异吲哚啉-2-基)环丁氧基)苯甲腈(400mg,0.80mmol),TBAF(1g,1.60mmol)溶于10mLTHF。室温搅拌过夜。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(150mg,0.35mmol)。收率43.9%。
LC/MS(ESI+)calcd for C25H24ClN2O2 +([M+H]+)m/z:419.2;found 419.1。
3.化合物3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-羧酸叔丁酯的合成
4-碘代-1H-吡唑(400mg,1.40mmol),N-Boc-3-碘代氮杂环丁烷(274mg,1.41mmol),K2CO3(292mg,2.12mmol)溶于5mL DMF。加热到85℃,搅拌过夜。冷却至室温。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-羧酸叔丁酯(380mg,1.08mmol)。收率77.0%。
4.化合物1-(氮杂环丁烷-3-基)-4-碘-1H-吡唑的合成
3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-羧酸叔丁酯(380mg,1.08mmol)溶于2mL二氯甲烷和2mL三氟乙酸。室温搅拌2h。浓缩,旋干,得到目标化合物(180mg,1.08mmol)。收率100.0%。
5.化合物2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-基)异吲哚啉-1,3-二酮的合成
1-(氮杂环丁烷-3-基)-4-碘-1H-吡唑(180.0mg,1.08mmol),2-(2,6-二氧哌啶-3-基)-5,6-二氟异吲哚啉-1,3-二酮(100mg,0.339mmol),DIEA(219mg,1.70mmol)溶于3mLDMSO。加热到130℃,搅拌3h。冷却至室温。加入水和乙酸乙酯萃取,有机层用饱和食盐水洗涤,干燥,旋干,过硅胶柱纯化。得到化合物2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-基)异吲哚啉-1,3-二酮(150mg,0.26mmol)。收率84.0%。
LC/MS(ESI+)calcd for C19H16FIN5O4 +([M+H]+)m/z:524.0;found 524.0。
6.化合物2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂环丁烷-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈的合成
氮气保护下,2-(2,6-二氧哌啶-3-基)-5-氟-6-(3-(4-碘-1H-吡唑-1-基)氮杂环丁烷-1-基)异吲哚啉-1,3-二酮(50mg,0.10mmol),2-氯-4-((1r,3r)-3-(5-乙炔基-1-氧异吲哚-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(40mg,0.10mmol),Pd(PPh3)2Cl2(8mg,0.01mmol),CuI(10mg,0.02mmol),0.3mL三乙胺加入到1mL甲苯中。加热到100℃,搅拌过夜。加冷却至室温。抽滤,用乙酸乙酯洗涤,干燥,旋干,过硅胶柱纯化。得到化合物(2-氯-4-((1r,3r)-3-(5-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂环丁烷-3-基)-1H-吡唑-4-基)乙炔基)-1-氧异吲哚啉-2-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(15mg,0.80mmol)。收率19.2%。LC/MS(ESI+)calcd for C44H38ClFN7O6 +([M+H]+)m/z:814.3;found 814.2.1H NMR(400MHz,CDCl3)δ8.05–7.98(m,1H),7.81(d,J=7.9Hz,1H),7.74(s,1H),7.64(s,1H),7.60–7.49(m,3H),7.41(d,J=12.2Hz,1H),7.09(d,J=7.2Hz,1H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.7,2.4Hz,1H),5.11–4.98(m,1H),4.97–4.88(m,1H),4.66(s,2H),3.81–3.69(m,3H),3.23(td,J=9.8,5.1Hz,1H),2.99–2.85(m,5H),2.85–2.70(m,2H),2.16–2.08(m,1H).1.46(s,6H),1.27(s,6H).
311:2-氯-4-((1r,4r)-4-(6-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈
1. 6-氟-4-甲基烟酸甲酯的合成
收率50%。LC/MS(ESI+)calcdfor:C8H8FNO2(M+H)m/z,170.1;found,170.1.
2. 4-(溴甲基)-6-氟烟酸甲酯的合成
收率45%。LC/MS(ESI+)calcdfor:C8H7BrFNO2(M+H+)m/z,170.1;found,170.1.
3.甲基4-(((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基)甲基)-6-氟烟酸甲酯的合成
收率55%。LC/MS(ESI+)calcd for:C21H21ClFN3O3(M+H+)m/z,170.1;found,170.1.
4. 2-氯-4-((1r,4r)-4-(6-氟-3-氧-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)
苯甲腈
收率40%。LC/MS(ESI+)calcd for:C20H17ClFN3O2(M+H+)m/z,386.1;found,386.1.
5. 2-氯-4-((1r,4r)-4-(6-(4-(羟甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈的合成
收率35%。LC/MS(ESI+)calcd for:C26H29ClN4O3(M+H+)m/z,481.2;found,481.2.
6. 2-氯-4-((1r,4r)-4-(6-(4-甲酰哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈的合成
收率52%。LC/MS(ESI+)calcd for:C26H27ClN4O3(M+H+)m/z,479.1;found,479.1.
7. 2-氯-4-((1r,4r)-4-(6-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈的合成
照前述实施例合成得到,收率41%。LC/MS(ESI+)calcdfor:C43H44ClFN8O6(M+H+)m/z,479.1;found,479.1.
312:2-氯-4-((1r,4r)-4-(6-((4-(2-(2,6-二氧哌啶-3-基)-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈
照前述实施例合成得到,收率33%。LC/MS(ESI+)calcdfor:C43H47ClN8O5(M+H+)m/z,791.3;found,791.3.
1H NMR(400MHz,CDCl3)δ8.64(d,J=0.9Hz,1H),8.06(s,1H),7.77(d,J=8.5Hz,1H),7.58(d,J=8.8Hz,1H),7.07-6.99(m,2H),6.92(s,1H),6.88(dd,J=8.8,2.4Hz,1H),6.67-6.63(m,1H),5.22(dd,J=13.3,5.2Hz,1H),4.46(t,J=14.4Hz,3H),4.34-4.23(m,5H),3.49(s,4H),3.04-2.73(m,8H),2.47(s,2H),2.35(dd,J=13.0,5.1Hz,1H),2.24(d,J=12.6Hz,3H),2.00(d,J=12.4Hz,5H),1.30(d,J=20.2Hz,6H).
313:2-氯-4-((1r,4r)-4-(6-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈
照前述实施例合成得到,收率30%。LC/MS(ESI+)calcdfor:C43H46ClFN8O5(M+H+)m/z,809.3;found,809.3.
314:2-氯-4-((1r,4r)-4-(6-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-3-氧代-1,3-二氢-2H-吡咯[3,4-c]吡啶-2-基)环己基)氧基)苯甲腈
照前述实施例合成得到,收率38%。LC/MS(ESI+)calcdfor:C43H45ClN8O6(M+H+)m/z,805.3;found,805.3.
315:2-氯-4-((1r,3r)-3-(2-((1'-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-[1,4'-二哌啶]-4-基)乙炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物4-((6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)乙炔基)-1,4'-联吡啶-1'-羧酸叔丁酯
将4-((1r,3r)-3-(2-溴-5-氧-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(500mg,1.06mmol),4-乙炔基-1,4'-二哌啶-1'-羧酸叔丁酯(619mg,2.12mmol)和三乙胺(0.5mL)溶入四氢呋喃(20mL)中,在氮气保护下加入四三苯基磷钯(127mg,0.11mmol)和碘化亚铜CuI(42mg,0.22mmol)。反应液室温搅拌过夜,然后旋干。硅胶柱层析(二氯甲烷:甲醇=100:1至25:1)纯化得到黄色固体4-((6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)乙炔基)-1,4'-联吡啶-1'-羧酸叔丁酯(280mg,0.41mmol),收率:39%。MS:calcd.forC39H49ClO5N4[M+H]+:686.3;found:686.4.
2.化合物4-((1r,3r)-3-(2-(1,4'-二哌啶-4-亚苯基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈
将4-((6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)乙炔基)-1,4'-联吡啶-1'-羧酸叔丁酯(140mg,0.12mmol)溶入二氯甲烷(4mL)中,在冰浴下加入三氟乙酸(2mL)。反应液在室温搅拌1小时,旋干得到4-((1r,3r)-3-(2-(1,4'-二哌啶-4-亚苯基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈粗品,直接用于下一步反应。MS:calcd.for C34H41ClO5N2[M+H]+:586.3;found:586.2.
3.化合物2-氯-4-((1r,3r)-3-(2-((1'-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)-1,4'-二哌啶-4-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将4-((1r,3r)-3-(2-(1,4'-二哌啶-4-亚苯基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)-2-氯苯甲腈(0.12mmol,粗品来源于上一步)和N,N-二异丙基乙胺(105mg,1.2mmol)溶于二甲亚砜(3mL)中,然后加入2-(2,6-二氧哌啶-3-基)-5-氟异吲哚-1,3-二酮(36mg,0.13mmol).反应液加热至110℃反应5小时。反应液用水(10mL)稀释,乙酸乙酯(10mL×3)萃取。有机相合并用饱和食盐水(10mL×3)洗涤,硫酸钠干燥,硅胶制备板薄层层析(二氯甲烷:甲醇=25:1)纯化得到黄色固体2-氯-4-((1r,3r)-3-(2-((1'-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚-5-基)-1,4'-二哌啶-4-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(15mg,0.018mmol),收率:9%。
MS:calcd.for C47H49ClO7N6[M+H]+:842.3;found:842.2.1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.03(d,J=7.9Hz,1H),7.88(d,J=8.6Hz,1H),7.61(t,J=15.8Hz,1H),7.48(t,J=30.7Hz,1H),7.37–7.05(m,3H),7.07(t,J=13.8Hz,1H),5.04(dd,J=12.7,5.2Hz,1H),4.78(s,2H),4.52(s,1H),4.28(d,J=31.9Hz,1H),4.07(d,J=11.9Hz,2H),3.01–2.89(m,2H),2.89–2.63(m,4H),2.89–2.63(m,2H),2.37–2.31(m,2H),2.00–1.97(m,2H),1.89–1.80(m,4H),1.63–1.61(m,2H),1.49–1.43(m,2H),1.38(s,6H),1.14(s,6H).MS:calcd.for C47H49ClO7N6[M+H]+:842.3;found:842.2.
以下化合物316-402用类似于315的方法合成得到。
316:2-氯-4-((1r,3r)-3-(2-((1'-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-1,4'-二哌啶-4-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.6Hz,1H),7.69(d,J=11.3Hz,1H),7.54(d,J=7.9Hz,1H),7.43(d,J=7.3Hz,1H),7.22(t,J=32.1Hz,1H),7.05(d,J=8.7Hz,1H),5.08(dd,J=12.5,5.2Hz,1H),4.79(s,2H),4.52(s,1H),4.32(s,1H),3.65(d,J=10.5Hz,2H),2.90–2.72(m,6H),2.63–2.55(m,2H),2.45–2.32(m,2H),2.03–1.97(m,2H),1.89–1.83(m,4H),1.63–1.58(m,4H),1.38(s,6H),1.14(s,6H).MS:calcd.for C13H11FO3N3[M+H]+:860.3;found:860.3.
317:2-氯-4-((1r,3r)-3-(2-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂苷-3-基)哌啶-4-基)乙炔基)-5-氧代-5H-吡咯啉[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.03(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.63(d,J=8.2Hz,1H),7.54(d,J=7.9Hz,1H),7.27(d,J=2.4Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),6.78(d,J=1.7Hz,1H),6.64(dd,J=8.3,1.9Hz,1H),5.04(dd,J=12.8,5.3Hz,1H),4.78(s,2H),4.52(s,1H),4.32(s,1H),4.09(t,J=7.6Hz,2H),3.92–3.66(m,2H),2.95–2.72(m,2H),2.72–2.57(m,2H),2.56–2.49(m,2H),2.40–2.24(m,1H),2.24–2.06(m,2H),2.04–1.81(m,3H),1.73–1.57(m,2H),1.38(s,6H),1.14(s,6H).MS:calcd.forC45H44ClN7O6[M+H]+:814.3;found:813.8.
318:2-氯-4-((1r,3r)-3-(2-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂苷-3-基)哌啶-4-基)乙炔基)-5-氧代-5H-吡咯啉[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.03(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.56(dd,J=17.6,9.5Hz,2H),7.27(d,J=2.4Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),6.90(d,J=7.7Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.78(s,2H),4.52(s,1H),4.32(s,1H),4.26–4.09(m,2H),4.04–3.84(m,2H),2.91–2.72(m,2H),2.68–2.58(m,2H),2.58–2.49(m,2H),2.42–2.22(m,1H),2.22–2.05(m,2H),2.04–1.87(m,3H),1.74–1.56(m,2H),1.38(s,6H),1.14(s,6H).MS:calcd.for C45H43ClFN7O6[M+H]+:832.3;found:831.7.
319:2-氯-4-((1r,3r)-3-(2-((1-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)壬二酸-3-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.04(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.64(d,J=8.5Hz,1H),7.57(d,J=7.9Hz,1H),7.39–7.15(m,3H),7.05(dd,J=8.8,2.4Hz,1H),5.04(dd,J=12.8,5.4Hz,1H),4.78(s,2H),4.52(s,1H),4.31(s,1H),3.83(d,J=12.9Hz,2H),3.60(s,2H),3.51–3.45(m,1H),3.18–2.99(m,4H),2.91–2.79(m,1H),2.61–2.50(m,2H),2.40–2.32(m,1H),2.01–1.96(m,1H),1.77–1.63(m,2H),1.36(s,6H),1.29–1.22(m,2H),1.14(s,6H).MS:calcd.for C45H44ClN7O6[M+H]+:814.3;found:814.3.
320:2-氯-4-((1r,3r)-3-(2-((1-(1-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)壬二酸-3-基)乙炔基)-5-氧代-5H-吡咯啉[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.05(d,J=7.1Hz,1H),7.89(d,J=8.8Hz,1H),7.69(d,J=11.6Hz,1H),7.58(d,J=7.6Hz,1H),7.43(d,J=7.7Hz,1H),7.27(d,J=2.2Hz,1H),7.05(dd,J=8.8,2.2Hz,1H),5.08(dd,J=12.4,5.3Hz,1H),4.79(s,2H),4.53(s,1H),4.32(s,1H),3.66–3.44(m,4H),3.14–3.07(m,1H),2.99–2.78(m,3H),2.63–2.52(m,2H),2.32–2.20(m,2H),2.06–1.90(m,2H),1.83–1.68(m,2H),1.49–1.22(m,8H),1.14(s,6H).MS:calcd.for C45H43ClFN7O6[M+H]+:832.3;found:832.3.
321:2-氯-4-((1r,3r)-3-(2-((1'-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-1,3'-二唑烷-3-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.05(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.60(d,J=18.3,8.1Hz,2H),7.27(d,J=2.4Hz,1H),7.05(d,J=8.8,2.4Hz,1H),6.77(d,J=1.9Hz,1H),6.64(d,J=8.4,2.0Hz,1H),5.03(dd,J=12.9,5.4Hz,1H),4.78(s,2H),4.52(s,1H),4.31(s,1H),4.03(t,J=7.9Hz,2H),3.81(dd,J=8.8,4.1Hz,2H),3.64–3.52(m,4H),3.25(d,J=15.6,9.3Hz,2H),2.95–2.75(m,1H),2.56–2.50(m,2H),2.08–1.90(m,1H),1.38(s,6H),1.14(s,6H).MS:calcd.for C43H40ClN7O6[M+H]+:786.3;found:785.8.
322:2-氯-4-((1r,3r)-3-(2-((1'-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-1,3'-二唑定-3-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.05(d,J=7.8Hz,1H),7.89(d,J=8.8Hz,1H),7.64–7.53(m,2H),7.27(d,J=1.9Hz,1H),7.05(d,J=8.6Hz,1H),6.90(d,J=7.7Hz,1H),5.04(d,J=12.8,5.5Hz,1H),4.78(s,2H),4.53(s,1H),4.31(s,1H),4.18(s,2H),3.94(s,2H),3.72–3.48(m,4H),3.25(d,J=6.5Hz,2H),2.85(t,J=12.8Hz,1H),2.69–2.53(m,2H),2.09–1.89(m,1H),1.38(s,6H),1.14(s,6H).MS:calcd.for C43H39ClFN7O6[M+H]+:804.3;found:804.2.
323:2-氯-4-((1r,4r)-4-(2-((1'-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-1,4'-二哌啶-4-基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)环己氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.01(d,J=7.9Hz,1H),7.85(d,J=8.8Hz,1H),7.63(d,J=8.5Hz,1H),7.51(d,J=7.9Hz,1H),7.39(d,J=2.3Hz,1H),7.30(s,1H),7.23(d,J=8.5Hz,1H),7.11(dd,J=8.8,2.3Hz,1H),5.04(dd,J=12.9,5.3Hz,1H),4.56(d,J=10.7Hz,1H),4.45(s,2H),4.14–4.01(m,3H),3.01–2.64(m,6H),2.56–2.50(m,3H),2.310-2.32(m,2H),2.13(d,J=10.3Hz,2H),2.03–1.93(m,1H),1.90–1.80(m,8H),1.71–1.40(m,6H).MS:calcd.for C45H44ClN7O6[M+H]+:814.3;found:814.3.
324:2-氯-4-((1r,4r)-4-(2-((1'-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-1,4'-二哌啶-4-基)乙炔基)-5-氧-5H-吡咯[3,4-b]吡啶-6(7H)-环己氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.04(d,J=7.9Hz,1H),7.87(d,J=8.8Hz,1H),7.71(d,J=11.4Hz,1H),7.55(d,J=7.9Hz,1H),7.45(d,J=7.4Hz,1H),7.41(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),5.10(dd,J=12.8,5.3Hz,1H),4.59(t,J=10.8Hz,1H),4.47(s,2H),4.22–4.08(m,1H),3.67(d,J=11.6Hz,2H),2.98–2.68(m,6H),2.63–2.52(m,2H),2.35–2.32(m,2H),2.16(d,J=10.7Hz,2H),2.08–1.76(m,10H),1.65–1.56(m,6H).MS:calcd.for C45H43ClFN7O6[M+H]+:832.3;found:832.2.
325:2-氯-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.90(d,J=8.7Hz,1H),7.68(d,J=8.5Hz,1H),7.54(d,J=7.9Hz,1H),7.33(s,1H),7.27(dd,J=12.2,5.5Hz,2H),7.06(dd,J=8.8,2.3Hz,1H),5.07(dd,J=12.9,5.3Hz,1H),4.79(s,2H),4.54(s,1H),4.33(s,1H),3.42(s,4H),2.96–2.81(m,1H),2.71–2.56(m,6H),2.36(d,J=24.4Hz,2H),2.17–1.95(m,3H),1.86(d,J=10.3Hz,2H),1.55–1.29(m,10H),1.16(s,6H).MS:calcd.for C47H48ClN7O6[M+H]+:842.3;found:842.2.
326:2-氯-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.02(d,J=7.9Hz,1H),7.89(d,J=8.7Hz,1H),7.71(d,J=11.5Hz,1H),7.52(d,J=7.9Hz,1H),7.42(d,J=7.4Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.78(s,2H),4.52(s,1H),4.31(s,1H),3.21(s,4H),2.85(dd,J=21.2,9.5Hz,1H),2.65(s,4H),2.60–2.55(m,2H),2.32–2.21(m,2H),2.11–1.96(m,3H),1.85(d,J=10.6Hz,2H),1.51–1.28(m,10H),1.14(s,6H).MS:calcd.for C47H47ClFN7O6[M+H]+:860.3;found:860.3.
327:2-氯-4-((1r,3r)-3-(2-((3-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.66(d,J=8.6Hz,1H),7.58(d,J=7.9Hz,1H),7.35–7.28(m,2H),7.24(dd,J=8.7,2.0Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),5.07(dd,J=12.9,5.4Hz,1H),4.81(s,2H),4.55(s,1H),4.34(s,1H),3.96(d,J=13.6Hz,2H),3.67(s,1H),3.27–3.17(m,1H),3.05(t,J=11.3Hz,2H),2.94–2.83(m,1H),2.72-2.74(m,1H),2.63–2.55(m,1H),2.34(d,J=7.9Hz,4H),2.19-2.21(m,2H),2.05–1.94(m,1H),1.84(d,J=10.8Hz,2H),1.39(d,J=12.4Hz,6H),1.32–1.23(m,2H),1.16(s,6H).MS:calcd.for C46H46ClN7O6[M+H]+:828.3;found:828.3.
328:2-氯-4-((1r,3r)-3-(2-((3-(1-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-22,44-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.02(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.63(d,J=8.5Hz,1H),7.51(d,J=7.9Hz,1H),7.35–7.23(m,2H),7.21(d,J=6.6Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.04(dd,J=12.9,5.4Hz,1H),4.76(s,2H),4.52(s,1H),4.31(s,1H),3.93(d,J=13.4Hz,2H),3.28–3.15(m,2H),3.03(t,J=11.2Hz,2H),2.96–2.79(m,2H),2.75–2.51(m,4H),2.04–1.84(m,3H),1.80(d,J=9.6Hz,2H),1.38(s,6H),1.33–1.22(m,2H),1.14(s,6H).MS:calcd.for C46H46ClN7O6[M+H]+:828.3;found:828.3.
329:2-氯-4-((1r,3r)-3-(2-((3-(1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.68(d,J=11.5Hz,1H),7.55(d,J=7.9Hz,1H),7.42(d,J=7.5Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.78(s,2H),4.53(s,1H),4.32(s,1H),3.69–3.58(m,1H),3.54(d,J=12.1Hz,2H),3.25(d,J=4.8Hz,1H),2.90–2.75(m,3H),2.61–2.53(m,1H),2.43–2.25(m,4H),2.16(d,J=9.4Hz,2H),2.03–1.96(m,1H),1.85(d,J=11.3Hz,2H),1.41–1.38(m,8H),1.14(s,6H).MS:calcd.for C46H45ClFN7O6[M+H]+:846.3;found:846.3.
330:2-氯-4-((1r,3r)-3-(2-((3-(1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.02(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.69(d,J=11.4Hz,1H),7.52(d,J=7.9Hz,1H),7.42(d,J=7.5Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.77(s,2H),4.53(s,1H),4.31(s,1H),3.54(d,J=12.2Hz,2H),3.28(s,2H),3.01–2.77(m,4H),2.72–2.50(m,4H),2.03–1.80(m,5H),1.47–1.23(m,8H),1.15(s,6H).MS:calcd.forC46H45ClFN7O6[M+H]+:846.3;found:846.2.
331:2-氯-4-((1r,3r)-3-(2-((3-((1-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)(甲基)氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.02(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.63(d,J=8.6Hz,1H),7.54(d,J=7.9Hz,1H),7.33–7.17(m,3H),7.05(dd,J=8.8,2.4Hz,1H),5.04(dd,J=12.9,5.4Hz,1H),4.77(s,2H),4.53(s,1H),4.31(s,1H),4.10(d,J=13.2Hz,2H),3.16–3.06(m,1H),2.95–2.78(m,4H),2.68–2.55(m,1H),2.55–2.50(m,4H),2.05–1.91(m,6H),1.63(d,J=11.8Hz,2H),1.48–1.44(m,2H),1.38(s,6H),1.14(s,6H).MS:calcd.for C47H48ClN7O6[M+H]+:842.3;found:842.3.
332:2-氯-4-((1r,3r)-3-(2-((3-((1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)(甲基)氨基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.02(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.68(d,J=11.4Hz,1H),7.54(d,J=7.9Hz,1H),7.42(d,J=7.5Hz,1H),7.26(d,J=2.4Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),3.64(d,J=11.4Hz,2H),3.18–3.06(m,1H),3.05–2.91(m,1H),2.86(t,J=12.4Hz,3H),2.62–2.49(m,5H),2.13–2.03(m,3H),2.03–1.90(m,3H),1.76–1.53(m,4H),1.38(s,6H),1.124(s,6H).MS:calcd.for C47H47ClFN7O6[M+H]+:860.3;found:860.3.
333:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)哌啶-4-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.69(d,J=11.5Hz,1H),7.54(d,J=7.9Hz,1H),7.42(d,J=7.5Hz,1H),7.28(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.09(dd,J=12.7,5.4Hz,1H),4.79(s,2H),4.53(s,1H),4.33(s,1H),3.59(d,J=12.3Hz,2H),3.29(s,1H),2.84(t,J=12.0Hz,3H),2.79–2.55(m,4H),2.18–2.16(m,4H),2.04–2.00(m,1H),1.90–1.88(m,2H),1.82–1.79(m,2H),1.74–1.61(m,3H),1.39(s,6H),1.28–1.20(m,2H),1.15(s,6H).MS:calcd.forC48H49ClFN7O6[M+H]+:874.3;found:874.0.
334:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)氮杂苷-3-基)甲基)哌啶-4-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-22,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.58(dd,J=11.4,9.7Hz,2H),7.29(d,J=2.4Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),6.90(d,J=7.6Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),4.81(s,2H),4.54(s,1H),4.34(s,1H),4.30–4.16(m,2H),3.91–3.79(m,2H),3.08–2.82(m,2H),2.74–2.65(m,3H),2.68–2.52(m,4H),2.38–2.12(m,2H),2.11–1.98(m,1H),1.96–1.82(m,2H),1.74–1.58(m,2H),1.41(s,6H),1.16(s,6H).MS:calcd.for C46H45ClFN7O6[M+H]+:846.3;found:845.8.
335:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)壬二酸-3-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.68(d,J=11.5Hz,1H),7.57(d,J=7.9Hz,1H),7.41(d,J=7.5Hz,1H),7.27(d,J=2.4Hz,1H),7.05(d,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.78(s,2H),4.53(s,1H),4.32(s,1H),3.64–3.42(m,5H),3.07(t,J=6.5Hz,2H),2.85(dd,J=22.0,10.1Hz,3H),2.67–2.51(m,2H),2.32(d,J=6.7Hz,2H),2.08–1.94(m,1H),1.77(d,J=11.2Hz,2H),1.47(s,1H)1.38(s,6H),1.30–1.25(m,2H),1.12(s,6H).MS:calcd.forC46H45ClFN7O6[M+H]+:846.3;found:845.8.
336:2-氯-4-((1r,3r)-3-(2-(3-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)-3-甲基-1-炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.73(d,J=11.4Hz,1H),7.58(d,J=7.9Hz,1H),7.45(d,J=7.4Hz,1H),7.27(d,J=2.4Hz,1H),7.04(d,J=8.8,2.4Hz,1H),5.09(dd,J=12.8,5.3Hz,1H),4.81(s,2H),4.51(s,1H),4.32(s,1H),2.93–2.76(m,5H),2.68–2.50(m,6H),2.07–1.95(m,1H),1.48(s,6H),1.38(s,6H),1.13(s,6H).MS:calcd.for C44H43ClFN7O6[M+H]+:820.3;found:819.7.
337:2-氯-4-((1r,3r)-3-(2-(1-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌啶-4-基氨基)-3-甲基-1-炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.8Hz,1H),7.89(d,J=8.8Hz,1H),7.69(d,J=11.5Hz,1H),7.53(d,J=7.8Hz,1H),7.42(d,J=7.4Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.79(s,2H),4.52(s,1H),4.32(s,1H),3.55(d,J=13.3Hz,2H),3.09–2.76(m,4H),2.62–2.49(m,2H),2.07–1.89(m,3H),1.59–1.45(m,2H),1.42–1.35(m,12H),1.14(s,6H).MS:calcd.forC45H45ClFN7O6[M+H]+:834.3;found:834.0.
338:2-氯-4-((1r,3r)-3-(2-((4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)苯基)乙炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.07(d,J=8.0Hz,1H),7.89(d,J=8.8Hz,1H),7.76(d,J=11.3Hz,1H),7.66(d,J=7.9Hz,1H),7.58–7.45(m,3H),7.28(d,J=2.4Hz,1H),7.05(d,J=9.0Hz,3H),5.10(dd,J=12.9,5.4Hz,1H),4.81(s,2H),4.54(s,1H),4.33(s,1H),3.45(s,4H),3.41(s,4H),2.87–2.65(m,1H),2.59–2.55(m,2H),2.08–2.00(m,1H),1.40(s,6H),1.15(s,6H).MS:calcd.for C47H41ClFN7O6[M+H]+:854.3;found:853.7.
339:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.01(d,J=7.7Hz,1H),7.88(d,J=8.7Hz,1H),7.61(d,J=12.3Hz,1H),7.51(d,J=7.6Hz,1H),7.26(d,J=2.2Hz,1H),7.13(d,J=7.2Hz,1H),7.04(d,J=8.8Hz,1H),5.04(d,J=12.7,5.3Hz,1H),4.77(s,2H),4.50(s,1H),4.31(s,1H),3.68(s,2H),3.19–3.08(m,2H),2.88(s,5H),2.70–2.53(m,5H),1.97(d,J=5.4Hz,2H),1.77–1.72(m,8H),1.36(s,6H),1.13(s,6H).MS:calcd.forC49H49ClFN7O6[M+H]+:886.3;found:885.7.
340:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.02(d,J=7.9Hz,1H),7.88(d,J=8.7Hz,1H),7.63(d,J=12.0Hz,1H),7.51(d,J=7.8Hz,1H),7.27(d,J=2.2Hz,1H),7.13(d,J=7.6Hz,1H),7.08–7.00(m,1H),5.06(d,J=12.8,5.2Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),3.69(s,2H),2.85(d,J=12.8Hz,5H),2.74–2.53(m,5H),2.07–2.00(s,8H),1.43(d,J=11.2Hz,2H),1.38(s,6H),1.26–1.21(m,2H),1.13(s,6H).MS:calcd.forC49H49ClFN7O6[M+H]+:886.3;found:886.2.
341:2-氯-4-((1r,3r)-3-(2-((4-(6-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.56(dd,J=19.2,9.5Hz,2H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),6.90(d,J=7.4Hz,1H),5.04(dd,J=12.8,5.3Hz,1H),4.80(s,2H),4.52(s,1H),4.33(s,1H),4.22(s,4H),3.30–3.01(m,4H),2.99–2.75(m,2H),2.72–2.49(m,1H),2.45–2.18(m,2H),2.12–1.90(m,2H),1.88–1.71(m,2H),1.70–1.41(m,4H),1.39(s,6H),1.14(s,6H).MS:calcd.for C48H47ClFN7O6[M+H]+:872.3;found:871.8.
342:2-氯-4-((1r,3r)-3-(2-((4-(6-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.02(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.55(dd,J=27.6,9.5Hz,2H),7.27(d,J=2.3Hz,1H),7.04(dd,J=8.8,2.3Hz,1H),6.90(d,J=7.7Hz,1H),5.04(dd,J=12.9,5.3Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),4.21(s,4H),3.29–3.03(m,4H),2.95–2.76(m,1H),2.67–2.51(m,2H),2.45–2.35(m,1H),2.09–1.85(m,4H),1.80–1.63(m,2H),1.52–1.38(m,2H),1.38(s,6H),1.14(s,6H),1.04–0.91(m,2H).MS:calcd.for C48H47ClFN7O6[M+H]+:872.3;found:871.8.
343:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.63(d,J=12.3Hz,1H),7.55(d,J=7.9Hz,1H),7.30(d,J=2.2Hz,1H),7.16(d,J=7.3Hz,1H),7.12–7.03(m,1H),5.07(dd,J=12.7,5.2Hz,1H),4.82(s,2H),4.61(s,1H),4.54(s,1H),4.35(s,1H),3.79(s,1H),3.61(s,1H),3.51(s,1H),3.16–3.08(m,1H),2.91–2.85(m,2H),2.62–2.53(m,1H),2.32–2.30(m,2H),2.04–2.00(m,2H),1.80–1.76(m,3H),1.72–1.48(m,7H),1.41(s,6H),1.16(s,6H).MS:calcd.for C48H47ClFN7O6[M+H]+:872.3;found:873.3.
344:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.00(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.61(d,J=12.4Hz,1H),7.49(d,J=7.9Hz,1H),7.26(d,J=2.3Hz,1H),7.13(d,J=7.7Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),5.05(dd,J=12.9,5.3Hz,1H),4.76(s,2H),4.58(s,1H),4.51(s,1H),4.31(s,1H),3.76(s,1H),3.61–3.58(m,1H),3.50–3.46(m,1H),3.10(d,J=7.7Hz,1H),2.87–2.64(m,1H),2.59–2.48(m,4H),2.27–2.24(m,1H),2.03–1.84(m,7H),1.49–1.32(m,8H),1.15–1.19(m,8H).MS:calcd.for C48H47ClFN7O6[M+H]+:872.3;found:872.0.
345:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)哌啶-4-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.7Hz,1H),7.63(d,J=8.5Hz,1H),7.54(d,J=7.9Hz,1H),7.36–7.21(m,2H),7.21(d,J=8.7Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.04(dd,J=13.0,5.4Hz,1H),4.79(s,2H),4.52(s,1H),4.32(s,1H),4.02(d,J=12.6Hz,2H),3.28(s,1H),2.97–2.82(m,3H),2.78–2.50(m,4H),2.13–2.08(m,4H),2.00–1.98(m,1H),1.87–1.83(m,2H),1.78–1.75(m,3H),1.65–1.62(m,2H),1.38(s,6H),1.14–1.07(m,8H).MS:calcd.for C48H50ClN7O6[M+H]+:856.4;found:856.0.
346:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)氮杂苷-3-基)甲基)哌啶-4-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯啉[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.61(d,J=8.3Hz,1H),7.54(d,J=7.8Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),6.76(d,J=2.0Hz,1H),6.69–6.49(m,1H),5.03(dd,J=13.0,5.4Hz,1H),4.79(s,2H),4.52(s,1H),4.32(s,1H),4.11(t,J=8.1Hz,2H),3.76–3.41(m,2H),3.22–2.84(m,2H),2.83–2.59(m,3H),2.59–2.49(m,4H),2.44–2.15(m,2H),2.13–1.95(m,1H),1.95–1.66(m,2H),1.76–1.46(m,2H),1.38(s,6H),1.14(s,6H).MS:calcd.forC46H46ClN7O6[M+H]+:828.3;found:827.8.
347:2-氯-4-((1r,3r)-3-(2-((1-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌啶-4-基)甲基)壬二酸-3-基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯啉[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.05(d,J=7.9Hz,1H),7.89(d,J=8.7Hz,1H),7.63(d,J=8.5Hz,1H),7.57(d,J=7.9Hz,1H),7.28(d,J=2.5Hz,2H),7.21(d,J=8.7,2.1Hz,1H),7.05(d,J=8.8,2.4Hz,1H),5.05(d,J=12.9,5.3Hz,1H),4.79(s,2H),4.53(s,1H),4.32(s,1H),4.01(d,J=13.3Hz,2H),3.61–3.47(m,3H),3.07(t,J=6.3Hz,2H),3.00–2.80(m,3H),2.63–2.53(m,2H),2.29(d,J=6.6Hz,2H),2.06–1.93(m,1H),1.74(d,J=11.5Hz,2H),1.57(s,1H),1.39(s,6H),1.20–1.03(m,8H).MS:calcd.forC46H46ClN7O6[M+H]+:828.3;found:827.8.
348:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),7.99(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.62(d,J=8.4Hz,1H),7.48(d,J=7.9Hz,1H),7.26(d,J=2.2Hz,1H),7.04(d,J=8.8,2.2Hz,1H),6.93(s,1H),6.84(d,J=8.6Hz,1H),5.02(d,J=12.9,5.3Hz,1H),4.76(s,2H),4.50(s,1H),4.30(s,1H),3.66(s,2H),3.28(d,J=7.0Hz,2H),2.90–2.85(m,5H),2.64–2.59(m,5H),2.08–1.92(m,2H),1.88–1.54(m,8H),1.36(s,6H),1.12(s,6H).MS:calcd.for C49H50ClN7O6[M+H]+:868.4;found:867.8.
349:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.6Hz,1H),7.91(d,J=8.8Hz,1H),7.67(d,J=9.0Hz,1H),7.53(d,J=7.8Hz,1H),7.29(d,J=2.2Hz,1H),7.07(d,J=8.8Hz,1H),6.96(s,1H),6.86(d,J=7.8Hz,1H),5.10–5.02(m,1H),4.79(s,2H),4.54(s,1H),4.33(s,1H),3.67(s,2H),3.49–3.38(m,3H),3.38(s,2H),2.93–2.73(m,4H),2.07–2.01(m,8H),1.50–1.45(m,2H),1.39(s,6H),1.28–1.24(m,3H),1.15(s,6H).MS:calcd.forC49H50ClN7O6[M+H]+:868.4;found:868.3.
350:2-氯-4-((1r,3r)-3-(2-((4-(6-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.03(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.62(d,J=8.3Hz,1H),7.54(d,J=7.9Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),6.77(d,J=1.8Hz,1H),6.62(dd,J=8.4,1.9Hz,1H),5.03(dd,J=12.9,5.5Hz,1H),4.80(s,2H),4.52(s,1H),4.33(s,1H),4.08(s,4H),3.25(s,4H),3.01–2.75(m,2H),2.68–2.52(m,2H),2.41–2.17(m,2H),2.09–1.91(m,2H),1.87–1.71(m,2H),1.69–1.41(m,4H),1.39(s,6H),1.14(s,6H).MS:calcd.for C48H48ClN7O6[M+H]+:854.3;found:853.8.
351:2-氯-4-((1r,3r)-3-(2-((4-(6-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.3]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.02(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.62(d,J=8.3Hz,1H),7.52(d,J=7.9Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),6.77(s,1H),6.63(d,J=8.3Hz,1H),5.03(dd,J=12.9,5.3Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),4.08(s,4H),3.30–3.23(m,4H),2.98–2.75(m,1H),2.67–2.50(m,2H),2.46–2.40(m,1H),2.21–1.85(m,4H),1.78–1.62(m,2H),1.56–1.39(m,2H),1.38(s,6H),1.14(s,6H),1.03–0.83(m,2H).MS:calcd.for C48H48ClN7O6[M+H]+:854.3;found:853.8.
352:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,,DMSO-d6)δ11.08(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.63(d,J=8.5Hz,1H),7.55(d,J=7.9Hz,1H),7.30(d,J=2.3Hz,1H),7.07(dd,J=8.8,2.4Hz,2H),6.89(s,1H),5.06(dd,J=12.9,5.3Hz,1H),4.82(s,2H),4.63(s,1H),4.54(s,1H),4.35(s,1H),3.85(s,1H),3.42(s,2H),3.11(d,J=12.1Hz,1H),2.89–2.67(m,2H),2.60–2.53(m,1H),2.34–2.32(m,1H),2.25–2.23(m,1H),2.18–2.02(m,2H),1.86–1.75(m,4H),1.74–1.50(m,6H),1.41(s,6H),1.16(s,6H).MS:calcd.for C48H48ClN7O6[M+H]+:854.3;found:854.3.
353:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.01(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.62(d,J=8.4Hz,1H),7.49(d,J=7.9Hz,1H),7.27(d,J=2.3Hz,1H),7.04(dd,J=8.8,2.3Hz,2H),6.87(s,1H),5.04(dd,J=13.0,5.3Hz,1H),4.76(s,2H),4.62(s,1H),4.51(s,1H),4.31(s,1H),3.81(s,1H),3.39(s,2H),3.07(s,1H),2.85–2.66(m,1H),2.59–2.51(m,2H),2.33–2.30(m,2H),2.21–2.19(m,1H),2.03–1.73(m,7H),1.45–1.34(m,8H),1.25–1.13(m,8H).MS:calcd.for C48H48ClN7O6[M+H]+:854.3;found:854.0.
354:2-氯-4-((1R,3r)-3-(2-((1s,3S)-3-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.03(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.66(d,J=8.5Hz,1H),7.54(s,1H),7.33(s,1H),7.29–7.24(m,2H),7.04(dd,J=8.8,2.4Hz,1H),5.05(dd,J=12.8,5.3Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),3.40(d,J=20.9Hz,4H),3.08–2.97(m,1H),2.93–2.79(m,1H),2.79–2.67(m,1H),2.66–2.49(m,4H),2.39(s,4H),2.05–1.96(m,3H),1.38(s,6H),1.14(s,6H).MS:calcd.forC45H44ClN7O6[M+H]+:814.3;found:814.0.
355:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.04(d,J=7.8Hz,1H),7.89(d,J=8.7Hz,1H),7.67(d,J=8.4Hz,1H),7.56(d,J=7.9Hz,1H),7.34(s,1H),7.29–7.24(m,2H),7.05(dd,J=8.8,2.5Hz,1H),5.05(dd,J=12.6,5.3Hz,1H),4.78(s,2H),4.53(s,1H),4.32(s,1H),3.43(s,4H),3.13–3.02(m,1H),2.91–2.78(m,1H),2.60–2.50(m,2H),2.41(s,4H),2.38–2.25(m,3H),2.27–2.13(m,2H),2.03–1.96(m,1H),1.38(s,6H),1.14(s,6H).MS:calcd.for C45H44ClN7O6[M+H]+:814.3;found:813.8.
356:2-氯-4-((1r,3r)-3-(2-((3-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.03(d,J=7.9Hz,1H),7.88(d,J=8.8Hz,1H),7.71(d,J=11.4Hz,1H),7.55(d,J=7.9Hz,1H),7.44(d,J=7.4Hz,1H),7.27(d,J=2.3Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),5.09(dd,J=12.8,5.4Hz,1H),4.78(s,2H),4.52(s,1H),4.31(s,1H),3.23(s,4H),3.09–2.98(m,1H),2.92–2.71(m,2H),2.61–2.49(m,4H),2.43(s,4H),2.07–1.92(m,3H),1.38(s,6H),1.14(s,6H).MS:calcd.forC45H43ClFN7O6[M+H]+:832.3;found:832.0.
357:2-氯-4-((1r,3r)-3-(2-((3-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.89(d,J=8.7Hz,1H),7.72(d,J=11.4Hz,1H),7.57(d,J=7.9Hz,1H),7.44(d,J=7.7Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),5.09(dd,J=12.8,5.4Hz,1H),4.79(s,2H),4.53(s,1H),4.32(s,1H),3.24(s,4H),3.17–3.07(m,1H),2.92–2.79(m,1H),2.62–2.49(m,2H),2.45–2.37(m,4H),2.36–2.26(m,3H),2.26–2.15(m,2H),2.05–1.97(m,1H),1.31(s,6H),1.18(s,6H).MS:calcd.for C45H43ClFN7O6[M+H]+:832.3;found:831.7.
358:2-氯-4-((1r,3r)-3-(2-((3-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.02(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.66(d,J=8.3Hz,1H),7.54(d,J=7.9Hz,1H),7.29(d,J=2.3Hz,1H),7.06(dd,J=8.8,2.3Hz,1H),6.96(s,1H),6.86(d,J=8.4Hz,1H),5.06(dd,J=12.9,5.3Hz,1H),4.78(s,2H),4.54(s,1H),4.32(s,1H),3.67(s,2H),3.31(s,3H),3.30–3.25(m,1H),3.11–2.82(m,5H),2.66–2.53(m,4H),2.46–2.39(m,2H),2.14–1.94(m,3H),1.39(s,6H),1.15(s,6H).MS:calcd.for C47H46ClN7O6[M+H]+:840.3;found:840.3.
359:2-氯-4-((1r,3r)-3-(2-((3-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.67(d,J=8.4Hz,1H),7.56(d,J=7.9Hz,1H),7.29(d,J=2.3Hz,1H),7.07(d,J=8.8,2.3Hz,1H),6.96(s,1H),6.87(d,J=8.8Hz,1H),5.07(d,J=12.9,5.4Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.67(s,2H),3.31(d,J=8.8Hz,2H),3.20(s,1H),2.99–2.78(m,3H),2.63–2.58(m,5H),2.48–2.42(m,2H),2.33(s,2H),2.20(s,2H),2.07–2.02(m,1H),1.40(s,6H),1.16(s,6H).MS:calcd.for C47H46ClN7O6[M+H]+:840.3;found:840.3.
360:2-氯-4-((1r,3r)-3-(2-((3-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.03(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.64(d,J=12.7Hz,1H),7.54(d,J=7.9Hz,1H),7.29(d,J=2.2Hz,1H),7.15(d,J=7.6Hz,1H),7.07(d,J=8.6Hz,1H),5.08(dd,J=12.5,5.1Hz,1H),4.78(s,2H),4.54(s,1H),4.33(s,1H),3.72(s,2H),3.32(s,4H),3.11–3.02(m,1H),2.89(s,4H),2.60(s,1H),2.48–2.40(m,5H),2.10–2.05(m,3H),1.40(s,6H),1.15(s,6H).MS:calcd.forC47H45ClFN7O6[M+H]+:858.3;found:857.8.
361:2-氯-4-((1r,3r)-3-(2-((3-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)六氢吡咯[3,4-c]吡咯-2(1H)-环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.65(d,J=12.5Hz,1H),7.57(d,J=7.9Hz,1H),7.29(d,J=2.2Hz,1H),7.16(d,J=7.4Hz,1H),7.07(d,J=8.8,2.2Hz,1H),5.08(dd,J=12.8,5.3Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.71(s,2H),3.32(s,2H),3.20(s,1H),2.88–2.80(m,3H),2.56–2.50(m,5H),2.49–2.41(m,2H),2.34(s,2H),2.21(s,2H),2.09–2.04(m,1H),1.40(s,6H),1.16(s,6H).MS:calcd.for C47H45ClFN7O6[M+H]+:858.3;found:858.3.
362:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.64(d,J=8.3Hz,1H),7.53(d,J=7.9Hz,1H),7.29(d,J=2.3Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),6.77(s,1H),6.65(dd,J=8.3,1.7Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.74(s,4H),3.25–3.16(m,2H),2.91–2.83(m,1H),2.68–2.57(m,2H),2.33(s,2H),2.09–1.98(m,4H),1.75(s,6H),1.45–1.35(m,11H),1.16(s,6H).MS:calcd.for C50H52ClN7O6[M+H]+:882.4;found:882.2.
363:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.04(d,J=8.0Hz,1H),7.91(d,J=8.8Hz,1H),7.59(t,J=12.2Hz,1H),7.53(d,J=7.9Hz,1H),7.29(d,J=2.2Hz,1H),7.07(dd,J=8.8,2.3Hz,1H),6.90(d,J=7.7Hz,1H),5.06(dd,J=12.8,5.3Hz,1H),4.80(s,2H),4.55(s,1H),4.34(s,1H),3.89(s,4H),3.32(s,2H),2.91–2.84(m,1H),2.67–2.56(m,2H),2.50–2.33(m,2H),2.09–2.00(m,4H),1.76–1.68(m,6H),1.50–1.22(m,11H),1.16(s,6H).MS:calcd.for C50H51ClFN7O6[M+H]+:900.4;found:900.3.
364:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[4.4]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1HNMR(400MHz,DMSO-d6)δ11.08(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.64(d,J=8.5Hz,1H),7.54(d,J=7.9Hz,1H),7.29(d,J=2.2Hz,1H),7.07(dd,J=8.8,2.3Hz,1H),6.90(s,1H),6.81(d,J=8.3Hz,1H),5.06(dd,J=13.0,5.3Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.46(s,2H),2.41–3.38(m,1H),3.32(s,2H),2.93–2.84(m,1H),2.60–2.54(m,6H),2.12–1.89(m,8H),1.77–1.75(m,2H),1.50–1.44(m,2H),1.40(s,6H),1.31–1.24(m,2H),1.16(s,6H).MS:calcd.for C50H52ClN7O6[M+H]+:882.4;found:882.3.
365:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[4.4]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.07(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.71–7.50(m,2H),7.30(d,J=2.4Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),6.90(s,1H),6.81(d,J=6.8Hz,1H),5.05(dd,J=12.6,5.4Hz,1H),4.82(s,2H),4.55(s,1H),4.35(s,1H),3.48(s,2H),3.42–3.35(m,1H),3.28–3.17(m,2H),2.92–2.84(m,1H),2.67–2.54(m,6H),2.02–1.91(m,2H),1.85–1.64(m,8H),1.41(s,6H),1.35–1.29(m,2H),1.23–1.20(m,2H),1.16(s,6H).MS:calcd.for C50H52ClN7O6[M+H]+:882.4;found:882.3.
366:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[4.4]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.02(d,J=7.8Hz,1H),7.88(d,J=8.7Hz,1H),7.58(d,J=12.5Hz,1H),7.51(d,J=7.9Hz,1H),7.27(d,J=2.1Hz,1H),7.03(t,J=9.1Hz,2H),5.04(dd,J=12.7,5.1Hz,1H),4.77(s,2H),4.52(s,1H),4.31(s,1H),3.60(s,2H),3.55–3.42(m,2H),3.28(s,1H),2.91–2.82(m,1H),2.62–2.54(m,6H),2.11–1.91(m,8H),1.78–1.70(m,2H),1.48–1.40(m,2H),1.38(s,6H),1.31–1.19(m,2H),1.14(s,6H).MS:calcd.for C50H51ClFN7O6[M+H]+:900.4;found:900.3.
367:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[4.4]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1HNMR(400MHz,DMSO-d6)δ11.09(s,1H),8.06(d,J=7.7Hz,1H),7.91(d,J=8.8Hz,1H),7.59(t,J=10.2Hz,2H),7.30(d,J=2.4Hz,1H),7.07(dd,J=13.5,4.8Hz,2H),5.06(dd,J=13.6,5.5Hz,1H),4.81(s,2H),4.54(s,1H),4.35(s,1H),3.64–3.60(m,2H),3.55–3.49(m,2H),3.31(s,1H),2.87–2.85(m,1H),2.67–2.54(m,6H),2.03–1.75(m,8H),1.68–1.63(m,2H),1.44(s,6H),1.34–1.28(m,2H),1.20–1.17(m,2H),1.16(s,6H).MS:calcd.forC50H51ClFN7O6[M+H]+:900.4;found:900.3.
368:2-氯-4-((1r,3r)-3-(2-((4-(6-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺环[3.4]辛基-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,CDCl3)δ8.04(d,J=7.9Hz,1H),7.99(s,1H),7.66(d,J=8.4Hz,1H),7.58(d,J=8.7Hz,1H),7.53–7.44(m,1H),7.00(d,J=2.3Hz,1H),6.93(s,1H),6.84(dd,J=8.7,2.4Hz,1H),6.67(d,J=9.3Hz,1H),4.93(dd,J=12.3,5.3Hz,1H),4.69(s,2H),4.46(s,1H),4.34–4.27(m,1H),3.56(s,2H),3.45(s,2H),3.23(s,3H),3.01–2.65(m,4H),2.27–2.23(m,2H),2.14–2.10(m,2H),2.02–1.97(m,2H),1.64–1.50(m,7H),1.46(s,6H),1.27(s,6H).MS:calcd.for C49H50ClN7O6[M+H]+:868.4;found:868.3.
369:2-氯-4-((1r,3r)-3-(2-((4-(6-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺环[3.4]辛基-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.65(d,J=8.4Hz,1H),7.54(d,J=7.9Hz,1H),7.29(d,J=2.4Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),6.90(s,1H),6.82(d,J=8.7Hz,1H),5.06(dd,J=13.0,5.4Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.53(s,2H),3.44(t,J=6.6Hz,2H),3.12(d,J=23.5Hz,3H),2.88–2.65(m,1H),2.56(dd,J=16.7,11.9Hz,4H),2.18–2.15(m,2H),2.18–2.02(m,4H),1.78–1.74(m,2H),1.47–1.33(m,8H),1.16(s,6H),1.10–1.00(m,2H).MS:calcd.for C49H50ClN7O6[M+H]+:868.4;found:868.3.
370:2-氯-4-((1r,3r)-3-(2-((4-(6-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.06(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.59(dd,J=18.4,10.2Hz,2H),7.30(d,J=2.4Hz,1H),7.12–7.00(m,2H),5.06(dd,J=12.9,5.5Hz,1H),4.82(s,2H),4.54(s,1H),4.35(s,1H),3.67(s,2H),3.57(s,2H),3.15–3.05(m,4H),3.01–2.82(m,2H),2.63–2.51(m,2H),2.12–2.10(m,2H),2.05–1.97(m,2H),1.85–1.81(m,2H),1.60–1.50(m,4H),1.45–1.30(m,8H),1.16(s,6H).MS:calcd.forC49H49ClFN7O6[M+H]+:886.3;found:886.3.
371:2-氯-4-((1r,3r)-3-(2-((4-(6-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.04(d,J=8.0Hz,1H),7.91(d,J=8.8Hz,1H),7.61(d,J=12.6Hz,1H),7.54(d,J=7.9Hz,1H),7.30(s,1H),7.06(t,J=8.6Hz,2H),5.07(dd,J=12.7,5.0Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.66(s,2H),3.57(s,2H),3.11–3.07(m,3H),2.88–2.88(m,1H),2.66–2.54(m,4H),211–2.01(m,6H),1.78–1.74(m,2H),1.50–1.39(m,8H),1.16(s,6H),1.10–1.00(m,2H).MS:calcd.forC49H49ClFN7O6[M+H]+:886.3;found:886.3.
372:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-6-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.04(d,J=8.1Hz,1H),7.89(d,J=8.7Hz,1H),7.63(d,J=8.9Hz,1H),7.55(d,J=8.2Hz,1H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.5Hz,1H),6.78(s,1H),6.63(s,1H),5.03(dd,J=12.7,5.4Hz,1H),4.80(s,2H),4.52(s,1H),4.33(s,1H),4.06–3.78(m,4H),2.99–2.69(m,4H),2.66–2.51(m,4H),2.15–1.94(m,3H),1.93–1.71(m,3H),1.65(m,6H),1.30(s,6H),1.18(s,6H).MS:calcd.forC49H50ClN7O6[M+H]+:868.4;found:868.3.
373:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-6-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.02(d,J=7.9Hz,1H),7.89(d,J=8.8Hz,1H),7.63(d,J=8.3Hz,1H),7.52(d,J=7.9Hz,1H),7.27(d,J=2.4Hz,1H),7.04(dd,J=8.8,2.4Hz,1H),6.77(s,1H),6.64(d,J=8.4Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),4.78(s,2H),4.52(s,1H),4.31(s,1H),4.13–3.68(m,4H),2.92–2.69(m,3H),2.67–2.50(m,4H),2.46–2.39(m,1H),2.16–1.81(m,8H),1.50–1.30(m,8H),1.32–1.22(m,2H),1.13(s,6H).MS:calcd.for C49H50ClN7O6[M+H]+:868.4;found:868.3.
374:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-6-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.07(s,1H),8.04(d,J=7.6Hz,1H),7.89(d,J=8.8Hz,1H),7.57(dd,J=15.4,9.5Hz,2H),7.27(d,J=2.4Hz,1H),7.05(dd,J=8.8,2.4Hz,1H),6.90(d,J=6.7Hz,1H),5.04(dd,J=12.9,5.4Hz,1H),4.80(s,2H),4.52(s,1H),4.33(s,1H),4.08(s,4H),2.98–2.74(m,3H),2.61–2.50(m,2H),2.44–2.31(m,3H),2.12–1.92(m,4H),1.89–1.77(m,2H),1.76–1.47(m,6H),1.39(s,6H),1.14(s,6H).MS:calcd.forC49H49ClFN7O6[M+H]+:886.3;found:886.3.
375:2-氯-4-((1r,3r)-3-(2-((4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,6-二氮杂螺[3.4]辛基-6-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.61(d,J=11.1Hz,1H),7.54(d,J=7.8Hz,1H),7.29(d,J=2.2Hz,1H),7.07(dd,J=8.8,2.2Hz,1H),6.93(d,J=7.7Hz,1H),5.07(dd,J=12.8,5.3Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),4.09(s,4H),2.94–2.70(m,3H),2.69–2.53(m,4H),2.46–2.39(m,1H),2.24–1.75(m,8H),1.56–1.29(m,8H),1.35–1.21(m,2H),1.16(s,6H).MS:calcd.for C49H49ClFN7O6[M+H]+:886.3;found:886.3.
376:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.06(d,J=7.7Hz,1H),7.91(d,J=8.8Hz,1H),7.63(d,J=8.4Hz,1H),7.57(d,J=7.9Hz,1H),7.30(d,J=2.3Hz,1H),7.07(dd,J=8.7,2.3Hz,1H),6.90(s,1H),6.81(d,J=8.4Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),4.82(s,2H),4.55(s,1H),4.35(s,1H),3.70–3.37(m,4H),3.05–2.82(m,2H),2.81–2.55(m,4H),2.37–1.92(m,6H),1.98–1.48(m,10H),1.41(s,6H),1.16(s,6H).MS:calcd.for C50H52ClN7O6[M+H]+:882.4;found:882.3.
377:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.64(d,J=8.4Hz,1H),7.54(d,J=7.9Hz,1H),7.29(d,J=2.2Hz,1H),7.07(dd,J=8.8,2.2Hz,1H),6.90(s,1H),6.81(d,J=8.4Hz,1H),5.06(dd,J=12.9,5.3Hz,1H),4.80(s,2H),4.54(s,1H),4.33(s,1H),3.53–3.37(m,4H),2.89–2.76(m,1H),2.63–2.52(m,6H),2.10–2.02(m,9H),1.78(s,2H),1.51–1.45(m,2H),1.40(s,6H),1.31–1.27(m,2H),1.16(s,6H).MS:calcd.for C50H52ClN7O6[M+H]+:882.4;found:882.3.
378:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.06(d,J=7.9Hz,1H),7.91(d,J=8.7Hz,1H),7.58(t,J=10.5Hz,2H),7.30(d,J=2.3Hz,1H),7.07(dd,J=14.2,5.1Hz,2H),5.06(dd,J=12.8,5.2Hz,1H),4.82(s,2H),4.55(s,1H),4.35(s,1H),3.70–3.48(m,4H),2.95–2.84(m,2H),2.70–2.55(m,4H),2.39–1.94(m,6H),1.90–1.60(m,10H),1.41(s,6H),1.16(s,6H).MS:calcd.for C50H51ClFN7O6[M+H]+:900.4;found:900.3.
379:2-氯-4-((1r,3r)-3-(2-((4-(7-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.04(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.57(dd,J=27.6,10.2Hz,2H),7.29(d,J=2.2Hz,1H),7.06(dd,J=13.2,4.8Hz,2H),5.07(dd,J=12.9,5.4Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.68–3.44(m,4H),2.94–2.83(m,1H),2.67–2.59(m,6H),2.17–1.69(m,11H),1.51–1.46(m,2H),1.40(s,6H),1.33–1.27(m,2H),1.16(s,6H).MS:calcd.for C50H51ClFN7O6[M+H]+:900.4;found:900.4.
380:2-氯-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ8.82(s,1H),7.92(d,J=8.7Hz,1H),7.74(d,J=11.5Hz,1H),7.45(d,J=7.3Hz,1H),7.28(t,J=12.3Hz,1H),7.08(dd,J=8.8,2.3Hz,1H),5.11(dd,J=12.8,5.4Hz,1H),4.82(d,J=19.3Hz,2H),4.63–4.49(m,1H),4.37(s,1H),3.24(s,4H),2.93–2.82(m,1H),2.77–2.54(m,6H),2.39–2.34(m,2H),2.12–1.97(m,3H),1.90–1.87(m,2H),1.59–1.32(m,10H),1.17(s,6H).MS:calcd.for C46H46ClFN8O6[M+H]+:861.3;found:861.3.
381:2-氯-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.82(s,1H),7.91(d,J=8.7Hz,1H),7.68(d,J=8.5Hz,1H),7.55–7.06(m,3H),7.08(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.8,5.3Hz,1H),4.84(s,2H),4.56(s,1H),4.37(s,1H),3.43(s,4H),2.92–2.84(m,1H),2.71–2.55(m,6H),2.49–2.33(m,2H),2.13–2.10(m,2H),2.03–1.99(m,1H),1.89–1.87(m,2H),1.55–1.46(m,2H),1.43(s,6H),1.38–1.23(m,2H),1.17(s,6H).MS:calcd.for C46H47ClN8O6[M+H]+:843.3;found:843.0.
382:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-吡咯[3,4-d]嘧啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲基亚砜基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(1054-1)的合成
将化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲硫基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(300mg,0.68mmol)溶入二氯甲烷(20mL),然后分批加入间氯过氧苯甲酸(300mg,1.70mmol)。反应液室温搅拌过夜。反应液用水(20mL)稀释,二氯甲烷(10mL×3)萃取。有机相合并用饱和亚硫酸钠(10mL×3)和饱和食盐水(10mL×3)洗涤,硫酸钠干燥,旋干得到白色固体化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲基亚砜基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(1054-1)粗品,直接用于下一步反应。MS:calcd.for C22H24ClO4N3S[M+H]+:459.1;found:459.1.
2.化合物4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-d]嘧啶-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(1054-2)的合成
将2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(2-(甲基亚砜基)-5-氧代-5H-吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(0.68mmol,粗品来源于上一步反应)和4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(193mg,0.68mmol)溶于二氯甲烷(20mL),然后加入N,N-二异丙基乙胺(310mg,2.38mmol)。反应液室温搅拌过夜。反应液用二氯甲烷(30mL)稀释,饱和食盐水(20mL×3)洗涤,无水硫酸钠干燥,硅胶柱层析(二氯甲烷:甲醇=100:1至20:1)纯化得到黄色固体4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-d]嘧啶-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(1054-2)(365mg,0.54mmol),两步收率:80%。MS:calcd.for C36H49ClO7N4[M+H]+:678.4;found:678.3.
3.化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(1054-3)的合成
将4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-d]嘧啶-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁酯(1054-2)(200mg,0.30mmol)溶入二氯甲烷(4mL),在冰浴下加入三氟乙酸(2mL)。反应液在室温搅拌1小时,旋干。然后溶入二氯甲烷(20mL)中,用饱和碳酸钠(10mL×3)和饱和食盐水(10mL×2)洗涤,无水硫酸钠干燥。旋干得到2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(1054-3)粗品,直接用于下一步反应。MS:calcd.for C31H41ClO7N2[M+H]+:578.3;found:578.3.
4.化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-吡咯[3,4-d]嘧啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-d]嘧啶-6(7H)-基)环丁氧基)苯甲腈(0.14mmol,粗品来源于上一步)和N,N-二异丙基乙胺(90mg,0.7mmol)溶于二甲亚砜(3mL)中,然后加入2-(2,6-二氧哌啶-3-基)-5,6-二氟异吲哚-1,3-二酮(60mg,0.20mmol)。反应液加热至110℃反应5小时。反应液用水(10mL)稀释,乙酸乙酯(10mL×3)萃取。有机相合并用饱和食盐水(10mL×3)洗涤,硫酸钠干燥,硅胶制备板薄层层析(二氯甲烷:甲醇=25:1)纯化得到黄色固体2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7二氢-吡咯[3,4-d]嘧啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(25mg,0.029mmol),收率:21%。MS:calcd.for C44H48ClFO9N6[M+H]+:852.3;found:852.3.
1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),8.63(s,1H),7.90(d,J=8.8Hz,1H),7.74(d,J=11.5Hz,1H),7.46(d,J=7.3Hz,1H),7.28(d,J=2.3Hz,1H),7.05(d,J=8.8,2.4Hz,1H),5.11(d,J=12.8,5.3Hz,1H),4.78(d,J=11.6Hz,2H),4.67(s,2H),4.49(s,1H),4.28(s,1H),3.26(s,4H),3.15–3.01(m,3H),2.95–2.82(m,1H),2.59–2.53(m,3H),2.23(d,J=6.6Hz,3H),2.10–1.78(m,5H),1.38(s,6H),1.19–1.00(m,8H).
383:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-吡咯[3,4-d]嘧啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,,DMSO-d6)δ11.10(s,1H),8.63(s,1H),7.90(d,J=8.7Hz,1H),7.69(d,J=8.5Hz,1H),7.35(s,1H),7.27(d,J=8.5Hz,2H),7.05(dd,J=8.8,2.0Hz,1H),5.08(d,J=12.8,5.3Hz,1H),4.78(d,J=11.9Hz,2H),4.66(s,2H),4.49(s,1H),4.28(s,1H),3.45(s,4H),3.09–2.82(m,4H),2.59–2.53(m,3H),2.21(d,J=6.9Hz,3H),2.10–1.76(m,5H),1.38(s,6H),1.14–1.06(m,8H).MS:calcd.for C44H48ClN9O6[M+H]+:834.3;found:832.9.
384:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1.化合物4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁基
将2-氯-4-((1r,3r)-3-(2-氯-5-氧-5H-吡咯[3,4-b]吡嗪-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(200mg,0.46mmol)和4-(哌啶-4-基甲基)哌嗪-1-羧酸叔丁酯(328mg,1.16mmol)溶于N,N-二甲基甲酰胺(5mL),然后碳酸钾(190mg,1.38mmol)。反应液升温至80℃搅拌过夜。反应液用水(10mL)稀释,乙酸乙酯(10mL×3)萃取。有机相合并用饱和食盐水(10mL×3)洗涤,硫酸钠干燥,硅胶柱层析(二氯甲烷:甲醇=100:1至20:1)纯化得到黄色固体4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁基(1056-1)(190mg,0.28mmol),收率:61%。MS:calcd.for C36H49ClO7N4[M+H]+:678.4;found:678.3.
2.化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-b]吡嗪-6(7H)-基)环丁氧基)苯甲腈
将4-((1-(6-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-5-氧代-6,7-二氢-5H-吡咯[3,4-b]吡嗪-2-基)哌啶-4-基)甲基)哌嗪-1-羧酸叔丁基(190mg,0.28mmol)溶入二氯甲烷(4mL),在冰浴下加入三氟乙酸(2mL)。反应液在室温搅拌1小时,旋干。然后溶入二氯甲烷(20mL)中,用饱和碳酸钠(10mL×3)和饱和食盐水(10mL×2)洗涤,无水硫酸钠干燥。旋干得到2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-b]吡嗪-6(7H)-基)环丁氧基)苯甲腈粗品,直接用于下一步反应。
MS:calcd.for C31H41ClO7N2[M+H]+:578.3;found:578.3.
3.化合物2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡嗪-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
将2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-(5-氧代-2-(4-(哌嗪-1-基甲基)哌啶-1-基)-5H吡咯[3,4-b]吡嗪-6(7H)-基)环丁氧基)苯甲腈(1056-2)(0.11mmol,粗品来源于上一步)和N,N-二异丙基乙胺(71mg,0.55mmol)溶于二甲亚砜(3mL)中,然后加入2-(2,6-二氧哌啶-3-基)-5,6-二氟异吲哚-1,3-二酮(SM-C-2)(42mg,0.15mmol)。反应液加热至110℃反应5小时。反应液用水(10mL)稀释,乙酸乙酯(10mL×3)萃取。有机相合并用饱和食盐水(10mL×3)洗涤,硫酸钠干燥,硅胶制备板薄层层析(二氯甲烷:甲醇=25:1)纯化得到黄色固体2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5H-吡咯[3,4-b]吡嗪-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈(19mg,0.022mmol),收率:20%。MS:calcd.for C44H48ClFO9N6[M+H]+:852.3;found:852.3.1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),8.53(s,1H),7.91(d,J=8.8Hz,1H),7.74(d,J=11.3Hz,1H),7.46(d,J=7.4Hz,1H),7.29(d,J=2.4Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),5.11(dd,J=12.6,4.9Hz,1H),4.68(s,2H),4.55(s,1H),4.43(d,J=12.7Hz,2H),4.32(s,1H),3.29–3.10(m,4H),3.04–2.78(m,3H),2.68–2.53(m,4H),2.44–2.18(m,4H),2.09–1.95(m,2H),1.92–1.62(m,4H),1.40(s,6H),1.17(s,6H).
385:2-氯-4-((1r,3r)-3-(2-(4-((4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.52(s,1H),7.91(d,J=8.7Hz,1H)7.69(d,J=8.5Hz,1H),7.35(s,1H),7.28(dd,J=10.1,5.8Hz,2H),7.07(dd,J=8.8,2.3Hz,1H),5.08(dd,J=12.7,5.4Hz,1H),4.68(s,2H),4.55(s,1H),4.43(d,J=12.4Hz,2H),4.32(s,1H),3.58–3.37(m,4H),3.04–2.80(m,3H),2.71–2.51(m,4H),2.40–2.15(m,4H),2.10–1.96(m,2H),1.94–1.71(m,4H),1.40(s,6H),1.17(s,6H).MS:calcd.for C44H48ClN9O6[M+H]+:834.3;found:834.3.
386:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.83(s,1H),7.92(d,J=8.8Hz,1H),7.63(d,J=12.4Hz,1H),7.30(d,J=2.4Hz,1H),7.16(d,J=7.5Hz,1H),7.08(dd,J=8.8,2.4Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.86(s,2H),4.61(s,1H),4.55(s,1H),4.38(s,1H),3.79(s,1H),3.62(d,J=8.9Hz,1H),3.52(d,J=8.7Hz,1H),3.13(d,J=8.7Hz,1H),2.95–2.63(m,2H),2.57–2.63(m,1H),2.46–2.38(m,2H),2.35–2.32(m,1H),2.04–1.94(m,1H),1.85–1.75(m,4H),1.74–1.49(m,6H),1.40(s,6H),1.17(s,6H).MS:calcd.forC47H46ClFN8O6[M+H]+:873.3;found:873.3.
387:2-氯-4-((1r,3r)-3-(2-((4-(5-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),8.81(d,J=8.5Hz,1H),7.91(d,J=8.8Hz,1H),7.64(d,J=12.4Hz,1H),7.30(d,J=2.3Hz,1H),7.16(d,J=6.7Hz,1H),7.07(dd,J=8.8,2.3Hz,1H),5.08(dd,J=12.9,5.2Hz,1H),4.88–4.83(m,2H),4.61–4.55(m,2H),4.36(s,1H),3.79(s,1H),3.63(s,1H),3.52(s,1H),3.14–3.11(m,1H),2.98–2.76(m,1H),2.67–2.54(m,4H),2.33–2.25(m,1H),2.03–1.85(m,7H),1.58–1.37(m,8H),1.23–1.16(m,8H).MS:calcd.for C47H46ClFN8O6[M+H]+:873.3;found:872.8.
388:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.83(s,1H),7.92(d,J=8.7Hz,1H),7.64(d,J=8.4Hz,1H),7.30(d,J=2.3Hz,1H),7.07(dt,J=12.0,6.0Hz,2H),6.88(s,1H),5.06(dd,J=12.9,5.4Hz,1H),4.86(s,2H),4.64(s,1H),4.55(s,1H),4.38(s,1H),3.86(s,1H),3.42(s,2H),3.12(d,J=8.4Hz,1H),3.03–2.81(m,2H),2.65–2.54(m,2H),2.35–2.25(m,2H),2.04–1.94(m,1H),1.93–1.78(m,4H),1.75–1.52(m,6H),1.43(s,6H),1.17(s,6H).MS:calcd.for C47H47ClN8O6[M+H]+:855.3;found:855.3.
389:2-氯-4-((1r,3r)-3-(2-((4-(5-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,5-二氮杂二环[2.2.1]庚烷-2-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),8.81(d,J=8.6Hz,1H),7.91(d,J=8.8Hz,1H),7.64(d,J=8.4Hz,1H),7.30(d,J=2.3Hz,1H),7.08–6.87(m,3H),5.06(dd,J=12.9,5.2Hz,1H),4.88–4.83(m,2H),4.60(d,J=34.6Hz,1H),4.55(s,1H),4.36(s,1H),3.84(s,1H),3.42(s,2H),3.11–3.09(m,1H),2.92–2.84(m,1H),2.67–2.55(m,4H),2.25–2.19(m,1H),2.02–1.81(m,7H),1.51–1.38(m,8H),1.27–1.16(m,8H).MS:calcd.forC47H47ClN8O6[M+H]+:855.3;found:854.9.
390:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),8.88(d,J=7.6Hz,1H),7.91(d,J=8.7Hz,1H),7.69(d,J=8.4Hz,1H),7.34(d,J=20.1Hz,1H),7.29(dd,J=9.7,5.6Hz,2H),7.08(dd,J=8.8,2.2Hz,1H),5.08(dd,J=12.9,5.3Hz,1H),4.90–4.85(m,2H),4.55(d,J=2.7Hz,1H),4.38(d,J=2.0Hz,1H),3.46(s,4H),3.15–3.12(m,1H),2.93–2.84(m,1H),2.68–2.55(m,2H),2.46–2.40(m,4H),2.39–2.33(m,3H),2.30–2.28(m,2H),2.04–2.01(m,1H),1.43(s,6H),1.17(s,6H).MS:calcd.for C44H43ClN8O6[M+H]+:815.3;found:815.3.
391:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡嗪-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),8.88(d,J=7.7Hz,1H),7.91(d,J=8.7Hz,1H),7.75(d,J=11.4Hz,1H),7.47(d,J=7.3Hz,1H),7.30(d,J=2.3Hz,1H),7.08(dd,J=8.8,2.3Hz,1H),5.11(dd,J=12.8,5.4Hz,1H),4.90–4.85(m,2H),4.56(d,J=2.7Hz,1H),4.38(s,1H),3.27(s,4H),3.20–3.15(m,1H),2.92–2.84(m,1H),2.68–2.52(m,2H),2.47–2.29(m,9H),2.08–2.02(m,1H),1.43(s,6H),1.18(s,6H).MS:calcd.forC44H42ClFN8O6[M+H]+:833.3;found:833.3.
392:4-((1R,3r)-3-(2-((1R,3r)-3-(4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲氧基苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.06(d,J=7.9Hz,1H),7.69(d,J=8.5Hz,1H),7.65(d,J=8.6Hz,1H),7.59(d,J=7.8Hz,1H),7.36(s,1H),7.28(d,J=8.0Hz,1H),6.69(d,J=2.0Hz,1H),6.60(dd,J=8.7,2.1Hz,1H),5.08(dd,J=12.9,5.5Hz,1H),4.82(s,2H),4.49(s,1H),4.34(s,1H),3.92(s,3H),3.64–3.34(m,4H),3.32–3.18(m,2H),3.17–3.03(m,1H),2.93–2.79(m,1H),2.71–2.54(m,2H),2.48–2.37(m,4H),2.38–2.31(m,1H),2.30–2.19(m,2H),2.08–1.95(m,1H),1.41(s,6H),1.18(s,6H).MS:calcd.forC46H47N7O7[M+H]+:810.4;found:810.3.
393:4-((1R,3r)-3-(2-((1R,3r)-3-(4-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲氧基苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.06(d,J=7.9Hz,1H),7.74(d,J=11.1Hz,1H),7.65(d,J=8.6Hz,1H),7.59(d,J=7.9Hz,1H),7.47(d,J=7.6Hz,1H),6.70(d,J=2.1Hz,1H),6.60(dd,J=8.7,2.2Hz,1H),5.11(dd,J=12.7,5.4Hz,1H),4.82(s,2H),4.49(s,1H),4.34(s,1H),3.92(s,3H),3.33–3.17(m,6H),3.17–3.10(m,1H),2.96–2.83(m,1H),2.74–2.54(m,2H),2.47–2.37(m,4H),2.36–2.33(m,1H),2.28–2.19(m,2H),2.07–1.99(m,1H),1.42(s,6H),1.18(s,6H).MS:calcd.for C46H46FN7O7[M+H]+:828.3;found:828.3.
394:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(1-(2,6-二氧哌啶-3-基)-6-氧代-1,6-二氢哒嗪-4-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-吡咯啉[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),8.06(d,J=7.9Hz,2H),7.91(d,J=8.8Hz,1H),7.58(d,J=7.9Hz,1H),7.30(d,J=2.4Hz,1H),7.07(d,J=8.8,2.4Hz,1H),5.87(d,J=2.7Hz,1H),5.59(d,J=12.1,5.2Hz,1H),4.81(s,2H),4.55(s,1H),4.34(s,1H),3.28(d,J=9.0Hz,4H),3.14–3.07(m,1H),2.92–2.80(m,1H),2.37(s,5H),2.23(s,2H),2.04–1.93(m,1H),1.69(s,6H),1.41(s,4H),1.16(s,6H).MS:calcd.for C41H43ClN8O5[M+H]+:763.3;found:763.3.
395:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(1-(2,6-二氧哌啶-3-基)-6-氧代-1,6-二氢嘧啶-4-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5H-吡咯[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.04(s,1H),8.17(s,1H),8.05(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,1H),7.58(d,J=7.9Hz,1H),7.29(d,J=2.4Hz,1H),7.07(dd,J=8.8,2.4Hz,1H),5.39(s,1H),5.32–5.07(m,1H),4.81(s,2H),4.55(s,1H),4.34(s,1H),3.51(s,4H),3.29–3.25(m,1H),3.14–3.05(m,1H),2.80–2.75(m,1H),2.60–2.53(m,2H),2.35–2.30(m,6H),2.24–2.20(m,2H),2.02–1.89(m,1H),1.41(s,6H),1.16(s,6H).MS:calcd.forC41H43ClN8O5[M+H]+:763.3;found:763.3.
396:2-氯-4-((1R,3r)-3-(2-((1R,3r)-3-(4-(3-(2,6-二氧哌啶-3-基)-4-氧代-3,4-二氢喹唑啉-7-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5H-吡咯啉[3,4-b]吡啶-6(7H)-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),8.22(s,1H),8.06(d,J=7.9Hz,1H),7.91(d,J=8.8Hz,2H),7.59(d,J=7.9Hz,1H),7.27(dd,J=24.2,5.8Hz,2H),7.07(dd,J=8.8,2.4Hz,1H),6.97(s,1H),4.81(s,2H),4.55(s,1H),4.34(s,1H),3.40(s,4H),3.14–3.05(m,1H),2.88–2.81(m,1H),2.67–2.57(m,2H),2.47–2.37(m,3H),2.36–2.27(m,1H),2.28–2.15(m,2H),2.14–2.05(m,1H),1.76(s,4H),1.36(s,6H),1.16(s,6H).MS:calcd.forC45H45ClN8O5[M+H]+:813.3;found:813.3.
397:2-溴-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO)δ11.12(s,1H),8.05(d,J=7.9Hz,1H),7.88(d,J=8.7Hz,1H),7.73(d,J=11.4Hz,1H),7.55(d,J=7.9Hz,1H),7.43(dd,J=13.7,4.8Hz,2H),7.10(dd,J=8.8,2.3Hz,1H),5.11(dd,J=12.7,5.4Hz,1H),4.80(s,2H),4.54(s,1H),4.34(s,1H),3.24(s,4H),2.92–2.84(m,1H),2.73–2.57(m,6H),2.46–2.31(m,2H),2.12–1.99(m,3H),1.88(d,J=10.0Hz,2H),1.49–1.35(m,10H),1.16(s,6H).MS:calcd.forC47H47BrFlN7O6[M+H]+:904.3;found:904.2.
398:2-溴-4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.88(d,J=8.7Hz,1H),7.68(d,J=8.5Hz,1H),7.54(d,J=7.9Hz,1H),7.42(d,J=2.4Hz,1H),7.34(s,1H),7.26(d,J=8.6Hz,1H),7.10(dd,J=8.8,2.4Hz,1H),5.07(dd,J=12.9,5.3Hz,1H),4.80(s,2H),4.54(s,1H),4.33(s,1H),3.42(s,4H),2.94–2.82(m,1H),2.72–2.52(m,6H),2.44–2.30(m,2H),2.12–1.98(m,3H),1.86(d,J=11.0Hz,2H),1.57–1.26(m,10H),1.16(s,6H).MS:calcd.for C47H48BrlN7O6[M+H]+:886.3;found:886.2.
399:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲氧基苯甲腈
1H NMR(400MHz,DMSO)δ11.12(s,1H),8.04(d,J=7.9Hz,1H),7.73(d,J=11.4Hz,1H),7.65(d,J=8.6Hz,1H),7.54(d,J=7.9Hz,1H),7.45(d,J=7.3Hz,1H),6.69(d,J=1.9Hz,1H),6.60(dd,J=8.7,2.0Hz,1H),5.11(dd,J=12.7,5.4Hz,1H),4.81(s,2H),4.49(s,1H),4.34(s,1H),3.92(s,3H),3.24(s,4H),2.96–2.81(m,1H),2.72–2.53(m,6H),2.44–2.30(m,2H),2.15–1.97(m,3H),1.87(d,J=9.3Hz,2H),1.57–1.28(m,10H),1.18(s,6H).MS:calcd.for C48H50FN7O7[M+H]+:856.4;found:856.3.
400:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲氧基苯甲腈
1H NMR(400MHz,DMSO)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.66(dd,J=12.1,8.6Hz,2H),7.54(d,J=7.9Hz,1H),7.34(s,1H),7.26(d,J=8.8Hz,1H),6.69(d,J=2.1Hz,1H),6.60(dd,J=8.7,2.1Hz,1H),5.08(dd,J=12.9,5.4Hz,1H),4.81(s,2H),4.49(s,1H),4.34(s,1H),3.92(s,3H),3.42(s,4H),2.97–2.80(m,1H),2.70–2.52(m,6H),2.45–2.22(m,2H),2.12–1.91(m,3H),1.86(d,J=10.5Hz,2H),1.66–1.21(m,10H),1.10(d,J=63.9Hz,6H).MS:calcd.for C48H51N7O7[M+H]+:838.4;found:838.3.
401:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-6-氟-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲基苯甲腈
1H NMR(400MHz,DMSO)δ11.10(s,1H),8.02(d,J=7.9Hz,1H),7.77–7.63(m,2H),7.52(d,J=7.8Hz,1H),7.42(d,J=7.3Hz,1H),6.96(s,1H),6.85(d,J=8.6Hz,1H),5.09(dd,J=12.8,5.5Hz,1H),4.78(s,2H),4.40(s,1H),4.30(s,1H),3.21(s,4H),2.92–2.80(m,1H),2.70–2.52(m,5H),2.40–2.24(m,2H),2.10–1.95(m,3H),1.85(d,J=9.8Hz,2H),1.59–1.23(m,9H),1.13(s,5H).MS:calcd.for C48H50FN7O6[M+H]+:840.4;found:840.3.
402:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-甲基苯甲腈
1H NMR(400MHz,DMSO)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.69(t,J=8.8Hz,2H),7.54(d,J=7.9Hz,1H),7.34(s,1H),7.26(d,J=9.0Hz,1H),6.99(d,J=2.2Hz,1H),6.87(dd,J=8.7,2.4Hz,1H),5.07(dd,J=12.9,5.4Hz,1H),4.80(s,2H),4.42(s,1H),4.32(s,1H),3.42(s,4H),2.94–2.82(m,1H),2.78–2.51(m,6H),2.47–2.28(m,5H),2.14–1.95(m,3H),1.86(d,J=10.7Hz,2H),1.55–1.27(m,10H),1.08(d,J=63.5Hz,6H).MS:calcd.for C48H51N7O6[M+H]+:822.4;found:822.3.
403:(2S,4R)-1-((S)-2-(2-((5-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基-3,3-d2)戊-4-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁醇基)-4-羟基-N-((S)-1-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
LC/MS(ESI+)calcd for C53H60D2ClN6O7S+([M+H]+)m/z:963.4;found 963.4.
404:(2S,4R)-1-((S)-2-(2-((6-(4-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)苯基)己-5-炔-1-基)氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
LC/MS(ESI+)calcd for C53H64BrN6O7S+([M+H]+)m/z:1007.4;found 1007.4.
1H NMR(400MHz,CDCl3)δ8.89(s,1H),7.70(d,J=8.1Hz,2H),7.56(d,J=8.7Hz,1H),7.46(t,J=9.5Hz,3H),7.38(d,J=17.8Hz,4H),7.22(d,J=8.6Hz,1H),7.15(d,J=2.2Hz,1H),6.85(dd,J=8.7,2.2Hz,1H),6.26(d,J=7.9Hz,1H),5.13–5.03(m,1H),4.76(t,J=7.8Hz,1H),4.54(d,J=8.5Hz,2H),4.19–4.10(m,2H),4.06(s,1H),3.97(q,J=15.4Hz,2H),3.59(t,J=6.4Hz,3H),2.60–2.47(m,6H),2.10–2.02(m,2H),1.82(dd,J=13.7,6.5Hz,2H),1.77–1.69(m,2H),1.47(d,J=6.9Hz,3H),1.27(s,6H),1.23(s,6H),1.07(s,9H).
405:(2R,4R)-1-((S)-2-(2-((5-((4-((1r,3r)-3-(4-(4H-1,2,4-三唑-1-基)苯氧基)-2,2,4,4-四甲基环丁基)氨甲酰)苯基)氨基)戊基)氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
LC/MS(ESI+)calcd for C53H70N9O7S+([M+H]+)m/z:976.5;found 976.4.
1H NMR(400MHz,CDCl3)δ8.67(s,1H),8.46(s,1H),8.08(s,1H),7.64(d,J=8.6Hz,2H),7.55(d,J=8.9Hz,2H),7.36(dd,J=18.2,8.3Hz,5H),7.24(d,J=9.0Hz,1H),6.97(t,J=12.3Hz,2H),6.63(d,J=8.2Hz,2H),6.12(d,J=8.1Hz,1H),5.12–4.98(m,1H),4.74(t,J=7.9Hz,1H),4.58(d,J=9.0Hz,1H),4.53(s,1H),4.13(t,J=10.5Hz,2H),4.06(d,J=8.2Hz,1H),3.71–3.41(m,4H),3.20(t,J=6.4Hz,2H),2.50(d,J=19.3Hz,4H),2.16–2.02(m,2H),1.72–1.65(m,4H),1.53(dd,J=15.1,9.7Hz,2H),1.45(s,3H),1.30–1.24(m,12H),1.05(d,J=18.9Hz,9H).
406:(2S,4R)-1-((S)-2-(2-((2-((5-(1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)氧基)戊基)氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-羧酰胺
LC/MS(ESI+)calcd for C53H70N9O7S+([M+H]+)m/z:981.4;found 981.2。
1H NMR(400MHz,CDCl3)δ8.80(s,1H),7.75(d,J=8.4Hz,1H),7.62–7.56(m,1H),7.47–7.33(m,6H),7.21(d,J=8.4Hz,1H),7.01–6.96(m,1H),6.91(s,1H),5.12–5.04(m,1H),4.75(t,J=7.8Hz,1H),4.61(d,J=4.4Hz,2H),4.54(d,J=8.6Hz,2H),4.30(d,J=7.2Hz,1H),4.14(d,J=11.4Hz,1H),4.04(t,J=6.3Hz,2H),3.95(q,J=15.4Hz,2H),3.67(s,1H),3.61(dd,J=11.3,3.5Hz,1H),3.56(t,J=6.5Hz,2H),2.54(d,J=10.1Hz,4H),2.08(d,J=13.5Hz,2H),1.91–1.82(m,2H),1.75–1.67(m,2H),1.63–1.57(m,2H),1.47(d,J=6.9Hz,3H),1.43(d,J=5.4Hz,6H),1.25(s,6H),1.08–1.04(m,9H).
407:(2S,4R)-1-((S)-2-(2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉啉-5基)-己-5-炔-1-基)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5基)-苯基)乙基)吡咯烷-2-甲酰胺
LC/MS(ESI+)calcd for C54H64ClN6O7S+([M+H]+)m/z:975.4;found 975.3。
1H NMR(400MHz,CDCl3)δ8.68(s,1H),7.76(d,J=7.9Hz,1H),7.57(d,J=8.7Hz,1H),7.48(d,J=8.1Hz,1H),7.46–7.33(m,5H),7.22(d,J=8.2Hz,1H),6.99(d,J=2.2Hz,1H),6.84(dd,J=8.8,2.2Hz,1H),5.14–5.01(m,1H),4.75(t,J=7.8Hz,1H),4.63(s,2H),4.54(d,J=8.6Hz,2H),4.39(s,1H),4.32(s,1H),4.15(d,J=11.8Hz,1H),3.97(q,J=15.4Hz,2H),3.61(d,J=6.9Hz,3H),2.89(s,1H),2.62–2.48(m,5H),2.14–1.94(m,2H),1.88–1.79(m,2H),1.77–1.70(m,2H),1.47(d,J=6.9Hz,3H),1.44(s,6H),1.25(s,6H),1.07(s,9H).
408:(2S,4R)-1-((S)-2-(2-((6-(2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)己基)氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)吡咯烷-2-甲酰胺
LC/MS(ESI+)calcd forC54H68ClN6O7S+m/z:979.5;found 979.4.
409:化合物(2S,4R)-1-((S)-2-(2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-哌啶-4-甲氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺合成
1.化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸叔丁酯的合成
氮气保护下,化合物4-((1r,3r)-3-(5-溴-1-异吲哚啉-2-基)-2,2,4,4-四甲基环丁基醚)-2-氯苯腈(200.0mg,0.42mmol),Pd2(dba)3(80.0mg,0.04mmol),BINAP(100.0mg,0.08mmol),2-(哌啶-4-甲氧基)乙酸叔丁酯(193.0mg,0.84mmol)和叔丁醇钠(81.0mg,0.84mmol)加入到4mL甲苯中,氮气置换3次。加热到110℃,搅拌12小时。冷却至室温,加入乙酸乙酯和饱和水萃取,有机层用饱和食盐水洗涤2次,无水硫酸钠干燥,旋干,硅胶柱层析纯化,得到无色油状液体2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸叔丁酯(90.0mg,0.14mmol)。收率:34.3%。
2.化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸的合成
化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸叔丁酯(90.0mg,0.14mmol)溶于到2mL三氟乙酸和1mL二氯甲烷,室温搅拌1h。溶剂真空旋干,得到化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸(90.0mg,0.14mmol)粗品,直接下一步反应。收率:100.0%。MS(ESI)m/e 513.0,515.0(M+H)+。
3.化合物(2S,4R)-1-((S)-2-(2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺的合成
化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸(25.0mg,0.04mmol),HATU(18.8mg,0.05mmol,)和三乙胺(29.0mg,0.20mmol),溶于到2mL DMF,再加入化合物(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(24.0mg,0.05mmol,),室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(2S,4R)-1-((S)-2-(2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酰胺)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺(22.0mg,0.02mmol)。收率:50.2%。
LC/MS(ESI+)calcd for C54H67ClN7O7S+([M+H]+)m/z:992.5;found 992.5.
1H NMR(400MHz,CDCl3)δ8.71(s,1H),7.79(s,1H),7.57(d,J=8.7Hz,1H),7.51–7.30(m,6H),7.12(d,J=8.3Hz,1H),6.99(d,J=2.3Hz,1H),6.84(dd,J=8.7,2.3Hz,1H),5.13–5.05(m,1H),4.78(t,J=7.9Hz,1H),4.67(d,J=20.7Hz,2H),4.60–4.49(m,2H),4.39(s,1H),4.31(s,1H),4.10(d,J=11.0Hz,1H),3.98(dd,J=33.0,15.2Hz,2H),3.82(d,J=11.1Hz,2H),3.62(dd,J=11.3,3.4Hz,1H),3.46(s,2H),3.07(s,2H),2.53(s,4H),2.12(s,1H),1.95(s,6H),1.49(d,J=6.9Hz,3H),1.44(s,6H),1.25(s,6H),1.07(s,9H).
410:(3R,5S)-1-((S)-2-(2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5哌啶-4-甲氧基)乙酰胺基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)酰胺基)吡咯烷-3-乙酸酯
化合物2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-哌啶-4-甲氧基)乙酸(25.0mg,0.04mmol),HATU(18.8mg,0.05mmol,)和三乙胺(29.0mg,0.20mmol),溶于到2mL DMF,再加入化合物(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(24.0mg,0.05mmol,),室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物化合物(3R,5S)-1-((S)-2-(2-((1-(2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5哌啶-4-甲氧基)乙酰胺基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)酰胺基)吡咯烷-3-乙酸酯(20mg,0.02mmol)。收率:50.1%。MS(ESI)m/e 513.0,515.0(M+H)+。
411:(2S,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-甲基环丁基)-1-氧异吲哚啉-5-氨基)戊烷)氧)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-酰胺的合成
1.2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-5-硝基-1-异吲哚啉-2-环丁基醚)苯腈的合成
化合物4-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁基醚)-2-氯苯腈(300.0mg,1.08mmol)和K2CO3(446.2mg,3.23mmol)加入到5mL DMF中,将2-溴甲基)-4-硝基苯甲酸甲脂(324.4mg,1.18mmol)分批加入到反应液中。室温搅拌1小时,TLC监测反应完全,生成一个新点。加入30mL乙酸乙酯和饱和食盐水萃取,有机层饱和食盐水洗涤2次,无水硫酸钠干燥,旋干,硅胶柱层析纯化。得到油状液体300mg,加入甲苯10mL,三乙胺1mL,加热回流,搅拌16h。溶剂旋干,加入少量石油醚和乙酸乙酯(10:1),固体抽滤,干燥,得到白色固体2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-5-硝基-1-异吲哚啉-2-环丁基醚)苯腈(250.0mg,0.56mmol)。收率:49.9%。
2.化合物4-((1r,3r)-3-(5-氨基)-1-异吲哚啉-2-基-2,2,4,4-四甲基环丁基醚-2-氯苯腈合成
化合物2-氯-4-((1r,3r)-2,2,4,4-四甲基-3-5-硝基-1-异吲哚啉-2-环丁基醚)苯腈(250.0mg,0.56mmol)溶于5mL乙酸,加入铁粉317.4mg,5.68mmol)。加热到50℃,搅拌1h。TLC监测反应完全,浓缩溶剂,加入乙酸乙酯,垫硅藻土抽滤,滤饼用乙酸乙酯洗涤和水洗涤。滤液加入饱和碳酸钾,调节pH至8-9,萃取,有机层用食盐水洗涤2次,无水硫酸钠干燥,旋干。得到化合物4-((1r,3r)-3-(5-氨基)-1-异吲哚啉-2-基-2,2,4,4-四甲基环丁基醚-2-氯苯腈(249.0mg,1.7mmol,)。收率:100%。
3.化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸叔丁酯的合成
化合物4-((1r,3r)-3-(5-氨基)-1-异吲哚啉-2-基-2,2,4,4-四甲基环丁基醚-2-氯苯腈(110.0mg,0.26mmol)和化合物2-((5-醛基戊烷)氧)乙酸叔丁酯(65.0mg,0.29mmol,)加入到2mL甲醇,再加入乙酸(32.0mg,0.53mmol)和氰基硼氢化钠(67.0mg,1.07mmol),室温搅拌16h。加入乙酸乙酯和饱和碳酸氢钠萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化。得到化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸叔丁酯(60.0mg,0.09mmol)。收率:36.6%。MS(ESI)m/e 513.0,515.0(M+H)+。
4.化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸的合成
化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸叔丁酯(60.0mg,0.09mmol)溶于到2mL三氟乙酸和1mL二氯甲烷,室温搅拌1h。溶剂真空旋干,得到化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸(70.0mg,0.09mmol)粗品,直接下一步反应。收率:100.0%。MS(ESI)m/e 513.0,515.0(M+H)+。
5.化合物(2S,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-甲基环丁基)-1-氧异吲哚啉-5-氨基)戊烷)氧)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺的合成
化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸(25.0mg,0.04mmol),HATU(18.8mg,0.05mmol,)和三乙胺(29.0mg,0.20mmol),溶于到2mL DMF,再加入化合物(2S,4R)-1-((S)-2-氨基-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-2-甲酰胺(24.0mg,0.05mmol,),室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(2S,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-甲基环丁氧)-1-异吲哚啉-5-氨基)戊烷)醚)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基乙酸酯-N-((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺(25mg,0.02mmol)。收率:54.2%。
LC/MS(ESI+)calcd for C53H67ClN7O7S+([M+H]+/2)m/z:980.5/2found 490.3.
1H NMR(400MHz,CDCl3)δ8.69(s,1H),7.73(d,J=8.0Hz,1H),7.70–7.60(m,1H),7.57(d,J=8.7Hz,1H),7.34(dd,J=20.7,8.0Hz,4H),7.23(d,J=9.4Hz,1H),6.99(d,J=1.9Hz,1H),6.97–6.87(m,1H),6.83(dd,J=8.7,2.0Hz,1H),5.00–4.89(m,1H),4.82(s,1H),4.67–4.58(m,2H),4.53(s,1H),4.38(s,1H),4.30(s,1H),4.09(d,J=11.4Hz,1H),3.95(dd,J=50.0,15.2Hz,2H),3.71–3.57(m,2H),3.52(s,1H),3.39–3.28(m,1H),3.26–3.15(m,1H),2.52(s,3H),2.36(s,1H),2.21(d,J=7.6Hz,1H),2.09–1.98(m,2H),1.81(s,2H),1.70(d,J=6.3Hz,2H),1.57–1.50(m,2H),1.43(t,J=3.9Hz,9H),1.29(d,J=2.4Hz,6H),1.09(s,9H).
412:(3R,5S)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯氧)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-氨基)戊烷)氧)乙酰胺基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)甲酰胺)吡咯烷-3-乙酸酯
化合物2-((5-((2-((1r,3r)-3-(3-氯-4-氰基苯醚)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)氧)乙酸(25.0mg,0.04mmol),HATU(18.8mg,0.05mmol,)和三乙胺(29.0mg,0.20mmol),溶于到2mL DMF,再加入化合物(3R,5S)-1-((S)-2-氨基-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-基)苯基)乙基)酰胺基)吡咯烷-3-乙酸酯(24.0mg,0.05mmol,),室温搅拌1h。加入乙酸乙酯和1N盐酸萃取,有机层用食盐水洗涤2次,干燥,旋干,硅胶柱层析纯化,得到化合物(3R,5S)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-氯氰基苯氧)-2,2,4,4-甲基环丁基)-1-氧异吲哚啉-5-氨基)戊烷)氧)乙酰胺基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)吡咯烷-2-甲酰胺(25mg,0.02mmol)。收率:54.2%。MS(ESI)m/e 1022(M+H)+。
413:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-恶二唑-5-甲酰胺
将3-溴-1,2,4-恶二唑-5-羧酸乙酯(221mg,1.0mmol)、4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(251mg,1.0mmol)溶解于5mL无水甲醇中,50℃加热过夜,冷却至室温,薄层色谱层析分离纯化,得到产品3-溴-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1,2,4-恶二唑-5-甲酰胺35mg,收率:8.2%,LC/MS(ESI+)called for C16H14BrClN4O3([M+H]+)m/e 425.0。
将4-哌啶甲醇(30mg,0.26mmol)溶于5mL DMF中加入碳酸钾(69mg,0.5mmol),加入3-溴-N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-1,2,4-恶二唑-5-甲酰胺(35mg,0.082mmol),80℃反应过夜,冷却至室温,加入10mL水,15mL乙酸乙酯萃取,有机层用水、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩干,薄层色谱层析分离纯化,得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-羟甲基)哌啶-1-基)-1,2,4-恶二唑-5-甲酰胺30mg,收率:79.5%,LC/MS(ESI+)called for C22H26ClN5O4([M+H]+)m/e 460.1。
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-羟甲基)哌啶-1-基)-1,2,4-恶二唑-5-甲酰胺(30mg,0.067mmol)溶解于5mL二氯甲烷,加入戴斯马丁氧化剂(64mg,0.15mmol),室温搅拌1h,过滤,滤液减压浓缩干,加入5mL二氯甲烷,加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚啉-1,3-二酮(26mg,0.066mmol),加入0.3滴乙酸,室温搅拌10min,加入三乙酰基硼氢化钠(106mg,0.5mmol),室温搅拌3h,水洗,饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-恶二唑-5-甲酰胺12mg,收率:21.7%,LC/MS(ESI+)called for C42H46ClN9O7([M+H]+)m/e 824.2,1H NMR(400MHz,CDCl3)δ8.06(s,1H),7.65(d,J=8.3Hz,1H),7.57(d,J=8.7Hz,1H),6.99(d,J=2.4Hz,1H),6.88–6.80(m,2H),6.79(d,J=2.0Hz,1H),6.52(dd,J=8.3,2.1Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.31(d,J=10.1Hz,1H),4.06(d,J=11.8Hz,2H),4.00(dd,J=7.2,3.6Hz,1H),3.78(s,4H),3.01–2.72(m,6H),2.45(s,2H),2.20(dd,J=8.9,5.1Hz,4H),2.07(s,3H),1.93(d,J=11.2Hz,4H),1.83(s,6H),1.66(s,2H),1.53–1.45(m,2H).
414:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-恶二唑-2-甲酰胺
将4-((1r,4r)-4-氨基环己基)氧基)-2-氯苯甲腈(10g,40mmol)溶于25mL二氯甲烷,加入三乙胺(1.21g,120mmol),冰水浴。将2-氯-2-氧乙酸甲酯(635mg,5.2mmol)溶于10mL二氯甲烷,滴加到反应体系中,滴加完毕,缓慢升温至室温,反应过夜,用饱和氯化铵水溶液洗涤2次,饱和食盐水洗涤一次,无水硫酸镁干燥,过滤,减压浓缩干,得到2-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基)-2-氧乙酸甲酯1.30g,收率:96.5%,LC/MS(ESI+)called for C16H17ClN2O4([M+H]+)m/e 337.1。
将2-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)氨基)-2-氧乙酸甲酯(1.30g,3.86mmol)溶于30mL无水乙醇,加入水合肼(706mg,14.1mmol),室温搅拌过夜,TLC检测反应完全,过滤,滤饼用10mL无水甲醇打浆一次,过滤,滤饼烘干,得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-肼基-2-氧乙酰胺1.25g,收率:96.2%,LC/MS(ESI+)called forC15H17ClN4O3([M+H]+)m/e 337.1。
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-肼基-2-氧乙酰胺(400mg,1.19mmol)溶于5mL四氢呋喃,加入CDI(211mg,1.5mmol),氩气保护,室温反应3小时,反应完全。加入4-哌啶甲醇(164mg,1.43mmol),反应液变溶清,室温反应过夜,减压弄过干,薄层色谱层析分离纯化,得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(2-(4-(羟甲基)哌啶-1-羰基)肼基)-2-氧乙酰胺511mg,收率:89.8%,LC/MS(ESI+)called for C22H28ClN5O5([M+H]+)m/e 478.2。
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-2-(2-(4-(羟甲基)哌啶-1-羰基)肼基)-2-氧乙酰胺(120mg,0.25mmol)溶于5mL二氯甲烷,加入三乙胺(125mg,1.25mmol),加入对甲苯磺酰氯(95mg,0.5mmol),室温反应过夜,饱和氯化铵水溶液洗涤2次,饱和食盐水洗涤一次,无水硫酸镁干燥,减压浓缩干,薄层色谱层析分离纯化,得到N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)-1,3,4-恶二唑-2-甲酰胺75mg,收率:65.2%,LC/MS(ESI+)called for C22H26ClN5O4([M+H]+)m/e 460.2。
将N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)-1,3,4-恶二唑-2-甲酰胺(40mg,0.087mmol)溶解于5mL二氯甲烷,加入戴斯马丁氧化剂(72mg,0.17mmol),室温搅拌1h,过滤,滤液减压浓缩干,加入5mL二氯甲烷,加入2-(2,6-二氧哌啶-3-基)-5-氟-6-(2,7-二氮杂螺[3.5]壬-2-基)异吲哚啉-1,3-二酮(33mg,0.087mmol),加入0.3滴乙酸,室温搅拌10min,加入三乙酰基硼氢化钠(106mg,0.5mmol),室温搅拌3h,水洗,饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩干,薄层色谱层析分离纯化,得到产品N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,3,4-恶二唑-2-甲酰胺30mg,收率:41.8%,LC/MS(ESI+)called for C42H46ClN9O7([M+H]+)m/e 824.3,1H NMR(400MHz,CDCl3)δ8.15–8.00(m,1H),7.72–7.61(m,1H),7.56(d,J=8.7Hz,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,2H),6.78(s,1H),6.59–6.47(m,1H),5.00–4.86(m,1H),4.34–4.25(m,1H),4.23–4.10(m,2H),4.05–3.94(m,1H),3.93–3.68(m,4H),3.69–3.48(m,1H),3.23–3.03(m,2H),2.97–2.65(m,5H),2.45–2.28(m,2H),2.17(d,J=10.7Hz,7H),1.97–1.79(m,3H),1.72–1.63(m,2H),1.61(s,5H),1.52–1.43(m,2H).
415:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-5-(4-((2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-恶二唑-3-甲酰胺
LC/MS(ESI+)called for C42H46ClN9O7([M+H]+)m/e 824.3,1H NMR(400MHz,CDCl3)δ8.06(s,1H),7.65(d,J=8.2Hz,1H),7.59–7.52(m,1H),6.99(d,J=2.4Hz,1H),6.84(dd,J=8.7,2.4Hz,1H),6.78(d,J=1.9Hz,1H),6.72(d,J=8.1Hz,1H),6.52(d,J=8.7Hz,1H),4.93(dd,J=12.2,5.3Hz,1H),4.29(d,J=10.1Hz,1H),4.23(d,J=14.0Hz,2H),4.03(dd,J=11.3,7.7Hz,1H),3.78(d,J=21.3Hz,4H),3.12(t,J=12.7Hz,2H),2.94–2.68(m,4H),2.38(s,3H),2.26–2.09(m,7H),1.87(s,4H),1.72–1.54(m,7H),1.49–1.40(m,2H).
416:N-((1r,4R)-4-(3-氯-4-氰基苯氧基)环己基)-1-((1r,4R)-4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬-7-基)甲基)环己基)-1H-吡唑-3-甲酰胺
1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),7.86(d,J=8.8Hz,1H),7.82(d,J=2.3Hz,1H),7.77(d,J=8.1Hz,1H),7.64(d,J=8.3Hz,1H),7.37(d,J=2.4Hz,1H),7.13(dd,J=8.8,2.4Hz,1H),6.78(s,1H),6.65(d,J=8.3Hz,1H),6.62(d,J=2.3Hz,1H),5.05(dd,J=12.8,5.2Hz,1H),4.50(s,1H),4.17(s,1H),2.86(dd,J=21.5,9.9Hz,1H),2.61–2.52(m,1H),2.33(s,4H),2.14–1.98(m,7H),1.76(s,6H),1.63–1.43(m,6H),1.23(s,6H),1.05(dd,J=12.2,5.2Hz,3H),0.84(d,J=7.0Hz,2H).LC/MS(ESI+)calcd for C44H49ClN8O6([M+H]+)m/z 821.38;found 821.3。
417:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-3-(4-((2-(2,6-二氧哌啶-3-基)-6-氟-1-氧异吲哚啉-5-基)-2,7-二氮杂螺环[3.5]壬-7-基)甲基)哌啶-1-基)-1,2,4-三嗪-6-甲酰胺
LC/MS(ESI+)Calcd for C43H48FClN10O5(M+H+)m/z,839.3;found839.3。1H NMR(400MHz,CDCl3)δ8.82(s,1H),8.09(s,1H),7.57(dd,J=8.5,5.4Hz,2H),7.40(d,J=11.3Hz,1H),7.00(d,J=2.4Hz,1H),6.85(dd,J=8.8,2.4Hz,1H),6.39(d,J=7.6Hz,1H),5.18(dd,J=13.2,5.2Hz,1H),4.33(dd,J=18.8,12.9Hz,2H),4.10–4.00(m,1H),3.83(s,3H),3.06(s,2H),2.92–2.75(m,2H),2.58(s,3H),2.45–2.27(m,4H),2.25–2.12(m,5H),2.09(s,2H),1.99(s,5H),1.72–1.62(m,2H),1.52–1.41(m,2H),1.32–1.20(m,3H).
418:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((7-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-2-基)甲基)-4-甲基哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C45H51ClN9O6[M+H]+m/z 848
419:N-((1r,4r)-4-(3-溴-4-氰基苯氧基)环己基)-3-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)-1,2,4-三氮唑-6-甲酰胺
LC/MS(ESI+)for C43H48BrN10O6[M+H]+m/z 879.
420:N-((1r,4r)-4-(4-氰基-3-三氟甲基-苯氧基)环己基)-3-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)-1,2,4-三氮嗪-6-甲酰胺
LC/MS(ESI+)for C44H48F3N10O6[M+H]+m/z 869.
421:N-((1r,4r)-4-(3-氯-4-氰苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)-1,2,4-三氮嗪-3-甲酰胺
LC/MS(ESI+)for C43H48ClN10O6[M+H]+m/z 835.
422:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)-4-甲基哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C45H51ClN9O6[M+H]+m/z 848.
423:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)-4-羟基哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C44H49ClN9O7[M+H]+m/z 850.
424:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)-4-氟哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C44H48ClFN9O6[M+H]+m/z 852.
425:2-氯-4-(((1r,4r)-4-(((6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-基)甲基)氨基)环己基)氧基)苯甲腈
LC/MS(ESI+)for C44H50ClN9O5[M+H]+m/z 820.
426:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)-4-氟哌啶-1-基)-1,2,4-三氮嗪-6-甲酰胺
LC/MS(ESI+)for C43H47ClFN10O6[M+H]+m/z 853.
427:N-((1r,4r)-4-((5-氯-6-氰基吡啶-3-基)氧基)环己基)-6-(4-((2-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C43H48ClN10O6[M+H]+m/z 835.
428:N-((1r,4r)-4-(3-氯-4-氰基苯氧基)环己基)-6-(4-((2-(6-(2,6-二氧哌啶-3-基)-5-氧-6,7-二氢-5H-吡咯[3,4-b]吡啶-2-基)-2,7-二氮杂螺[3.5]壬烷-7-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺
LC/MS(ESI+)for C43H50ClN10O5[M+H]+m/z 821.
429:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-氘代甲氧基苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.66(dd,J=13.5,8.6Hz,2H),7.54(d,J=7.9Hz,1H),7.34(s,1H),7.26(d,J=8.6Hz,1H),6.69(d,J=2.1Hz,1H),6.60(dd,J=8.7,2.2Hz,1H),5.07(dd,J=12.9,5.3Hz,1H),4.81(s,2H),4.49(s,1H),4.33(s,1H),3.42(s,4H),2.97–2.78(m,1H),2.74–2.52(m,6H),2.46–2.33(m,2H),2.18–1.94(m,3H),1.87(d,J=10.1Hz,2H),1.55–1.28(m,10H),1.18(s,6H).MS:calcd.forC48H48D3N7O7[M+H]+:841.40;found:841.3.
430:4-((1R,3r)-3-(2-((1R,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧代-5,7-二氢-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)-2-乙氧基苯甲腈
1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),8.04(d,J=7.9Hz,1H),7.68(d,J=8.5Hz,1H),7.64(d,J=8.6Hz,1H),7.54(d,J=7.9Hz,1H),7.34(s,1H),7.26(d,J=8.7Hz,1H),6.67(d,J=2.0Hz,1H),6.59(d,J=8.8Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.80(s,2H),4.48(s,1H),4.33(s,1H),4.19(q,J=7.0Hz,2H),3.42(s,4H),2.87–2.65(m,1H),2.63–2.53(m,6H),2.41–2.35(m,1H),2.15–1.98(m,4H),1.86(d,J=10.3Hz,2H),1.47–1.36(m,13H),1.17(s,6H).MS:calcd.for C49H53N7O7[M+H]+:852.4;found:852.2.
431:2-溴-4-((1R,3r)-3-(2-((1r,3R)-3-(4-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)哌嗪-1-基)环丁基)乙炔基)-5-氧代-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
1H NMR(400MHz,DMSO-d6)δ8.06(d,J=7.9Hz,1H),7.88(d,J=8.7Hz,1H),7.69(d,J=8.5Hz,1H),7.59(d,J=7.9Hz,1H),7.42(d,J=2.4Hz,1H),7.36(s,1H),7.28(d,J=8.8Hz,1H),7.10(dd,J=8.8,2.4Hz,1H),5.08(dd,J=12.9,5.3Hz,1H),4.81(s,2H),4.54(s,1H),4.34(s,1H),3.46(s,4H),3.28-3.20(m,1H),3.18–3.05(m,1H),2.96–2.81(m,1H),2.66–2.53(m,2H),2.43-2.35(m,6H),2.28–2.24(m,2H),2.08–1.94(m,1H),1.40(s,6H),1.16(s,6H).MS:calcd.for C45H44BrN7O6[M+H]+:858.3;found:858.2.
432:2-氯-4-((1R,3r)-3-(2-(((1r,4R)-4-(4-(2-(2,6-二氧哌啶-3-基)-1-氧异吲哚啉-5-基)哌嗪-1-基)环己基)乙炔基)-5-氧-5,7-二氢-6H-吡咯[3,4-b]吡啶-6-基)-2,2,4,4-四甲基环丁氧基)苯甲腈
LCMS for C47H51ClN7O5[M+H]+m/z 828.
433:(2S,4R)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-氧异吲哚啉-5-基)氨基)戊烷基)氧基)乙酰氨基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑5-基)苯基)乙基)吡咯烷基-2-甲酰胺
LCMS for C53H67BrN7O5S[M+H]+m/z 1024.
434:(3R,5S)-1-((S)-2-(2-((5-((2-((1r,3r)-3-(3-溴-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-1-异吲哚啉-5-氨基)戊烷)醚)乙酰胺基)-3,3-二甲基丁酰基)-5-(((S)-1-(4-(4-甲基噻唑-5-苯基)乙基)氨基甲酰)吡咯烷基-3-乙酸酯
1H NMR(400MHz,DMSO–d6)δ8.98(s,1H),8.46(d,J=7.7Hz,1H),7.86(d,J=8.7Hz,1H),7.39(m,6H),7.08(dd,J=8.8,2.4Hz,1H),6.62(m,2H),6.33(t,J=5.2Hz,1H),5.20(s,1H),4.89(m,1H),4.61(s,2H),4.49(s,1H),4.45(m,2H),4.25(s,1H),3.91(m,3H),3.77(dd,J=11.8,3.9Hz,1H),3.50(dd,J=15.0,8.6Hz,2H),3.30(s,1H),3.08(m,2H),2.45(s,3H),2.27(m,1H),2.00(s,3H),1.96(m,1H),1.59(dd,J=13.8,7.0Hz,4H),1.45(m,2H),1.36(s,6H),1.34(s,3H),1.12(s,6H),0.95(s,9H).LC/MS(ESI+)calcd for C55H68BrN7O8S([M+H]+)m/z 1066.4;found 1066.4.
435:(2R,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲氧基)苯甲酰胺基)己基-5-炔-1-)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-)苯基)乙基)四氢吡咯-2-酰胺
将2-氯-4-((1r,3r)-3-(甲氧胺基)-2,2,4,4-四甲基环丁基)苯腈溶于甲苯中,加入对碘苯甲酸甲酯,吡啶,60℃下搅拌过夜。反应完毕后,加入0.5N烯盐酸进行淬灭,乙酸乙酯萃取,饱和碳酸氢钠洗,水洗,饱和氯化钠洗,干燥,浓缩,柱层析即得产品(100mg,产率12%)。LC/MS(ESI+)calcd for C23H25ClIN2O3([M+H]+)m/z 539.1;found 539.1.
第二步:合成中间体:2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲氧基)苯甲酰胺基)己基-5-炔-1-)氧基)乙酸叔丁酯
将上述所得产品N-((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)-4-碘-N-甲氧基苯甲酰胺,以及2-(己基-5-炔-1-氧基)乙酸叔丁酯,Pd(dppf)Cl2,CuI,Et3N溶于甲苯中,110℃下搅拌24小时。待反应体系冷却,通过硅藻土过滤,乙酸乙酯淋洗,稀盐酸洗,饱和碳酸氢钠洗,水洗,饱和氯化钠洗,干燥,浓缩,柱层析即得产品(56mg,产率49%)。LC/MS(ESI+)calcd for C35H44ClN2O6([M+H]+)m/z 623.3;found 623.3.
第三步:合成最终产物:(2R,4R)-1-((S)-2-(2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲氧基)苯甲酰胺基)己基-5-炔-1-)氧基)乙酰胺基)-3,3-二甲基丁酰基)-4-羟基-N-((S)-1-(4-(4-甲基噻唑-5-)苯基)乙基)四氢吡咯-2-酰胺
将上述所得产品2-((6-(4-(((1r,3r)-3-(3-氯-4-氰基苯氧基)-2,2,4,4-四甲基环丁基)(甲氧基)苯甲酰胺基)己基-5-炔-1-)氧基)乙酸叔丁酯溶于二氯甲烷中,加入三氟乙酸,室温搅拌过夜。TLC监测原料反应完全,浓缩反应液。将浓缩所得物溶于DMF中,加入过量DIEA,加入VHL(OH),HATU,室温搅拌4个小时。反应完毕后,加入0.5N烯盐酸进行淬灭,乙酸乙酯萃取,饱和碳酸氢钠洗,水洗,饱和氯化钠洗,干燥,浓缩,柱层析即得产品(9mg,产率47%)。1H NMR(400MHz,DMSO–d6)δ8.96(s,1H),8.44(d,J=7.7Hz,1H),7.84(d,J=8.7Hz,1H),7.41(m,7H),7.08(dd,J=8.8,2.4Hz,1H),6.64(m,2H),6.33(t,J=5.2Hz,1H),5.19(s,1H),4.88(m,1H),4.52(s,1H),4.25(s,1H),3.88(m,3H),3.78(dd,J=11.8,3.9Hz,1H),3.67(s,3H),3.58(m,1H),3.52(dd,J=15.0,8.6Hz,2H),3.07(m,2H),2.26(m,1H),2.00(s,3H),1.95(m,1H),1.58(dd,J=13.8,7.0Hz,4H),1.44(m,2H),1.35(s,6H),1.33(s,3H),1.11(s,6H),0.94(s,9H).LC/MS(ESI+)calcd for C54H66ClN6O8S([M+H]+)m/z 993.4;found993.4.
以下通过实验例证明本发明的有益效果。
实验例1本发明化合物对前列腺癌细胞的增殖抑制活性
(1)实验材料和仪器:
LNCaP/AR cell line(由四川康城生物科技有限公司提供)
胎牛血清FBS(Gibco,Cat.No.10099-141)
0.01M PBS(Biosharp,Cat.No.162262)
RIPM1640培养基(Hyclone,Cat.No.308090.01)
青-链霉素Penicillin-Streptomycin(Hyclone,Cat.No.SV30010)
Cell counting kit-8试剂盒(Signalway Antibody,Cat.No.CP002)
二甲基亚砜DMSO(Sigma,Cat.No.D5879)
离心管Centrifuge Tube,15ml(Excell Bio,Cat.No.CS015-0001)
细胞培养皿Cell Culture Dish,(Excell Bio,Cat.No.CS016-0128)
细胞培养板96-well cell culture cluster(Corning,Cat.No.3599)
酶标仪(Thermo Multiskan MK3型)
(2)实验方法:
a.缓冲液配制
细胞培养液:RIPM1640培养基,10%FBS,1%Pen Strep;
PBS缓冲液:PBS粉剂溶于2L超纯水中,灭菌。
b.实验步骤:
1)LNCaP/AR细胞用细胞培养液传代培养,取生长状态良好的细胞接种于96孔板,每孔80μL,每孔细胞数为1000,于37℃,5%CO2细胞孵育箱中培养过夜。
2)将待测化合物用二甲基亚砜(DMSO)配置成10mM的储存液。临用前再用DMSO稀释3倍,再按3倍梯度稀释,得到9个浓度梯度,再用培养液将各浓度的化合物稀释200倍(以此保证培养体系中DMSO浓度为0.1%),每个浓度做2个孔重复。取20μL稀释好的化合物加到细胞培养孔(终浓度为10μM,3.3μM,1.1μM…),轻轻振荡混匀。另外设置3个只加细胞的阴性对照孔和3个只加培养液的空白对照孔(6孔各加20μL培养液稀释200倍的DMSO)。
c.结果检测:
1)培养6天后,每孔加10μL CCK-8,于37℃,5%CO2细胞孵育箱中继续培养1小时。
2)用多功能酶标仪在450nm处测定吸光度(OD值)。
3)数据用软件GraphPad Prism5中Dose-response-inhibition方程分析,得出IC50值。
(3)实验结果:
本发明实施例各化合物对LNCap/AR细胞增殖的抑制活性结果如表1所示。可以看出,本发明化合物能够有效抑制对LNCap/AR细胞增殖,大部分化合物对LNCap/AR细胞的IC50值甚至<0.1μM。
表1.本发明化合物对LNCap/AR细胞增殖抑制活性
A:<0.1μM,B:0.1-0.5μM,C:0.5-1μM,D:>1μM.
实验例2用酶联免疫吸附检测法(ELISA)分析本发明化合物对雄激素受体(AR)蛋白的降解活性
实验材料:
前列腺癌VCaP细胞(ATCC,CRL-2876)
活性炭处理过的胎牛血清Charcoal stripped FBS(Gibco,Cat.No.12676-029)
Metribolone(R1881)(Macklin Biochemical,CAS.No.965-93-5)
磷酸盐缓冲液0.01M PBS(Biosharp,Cat.No.162262)
DMEM/HIGH GLUCOSE(Hyclone,Cat.No.SH30243.01)
Penicillin-Streptomycin(Hyclone,Cat.No.SV30010)
二甲亚砜DMSO(Sigma,Cat.No.D5879)
离心管Centrifuge Tube,15ml(Excell Bio,Cat.No.CS015-0001)
细胞培养皿Cell Culture Dish,(Excell Bio,Cat.No.CS016-0128)
酶联免疫吸附检测法检测盒
Total Androgen Receptor SandwichELISA Kit(CST,Cat.No.12850C)
实验方法:
1.缓冲液配制
2.实验步骤:
1)VCaP细胞用细胞培养液传代培养后,取生长状态良好的细胞接种于96孔板,每孔80ul,每孔细胞数为30000个,于37℃,5%CO2细胞孵育箱中培养3天,待细胞贴壁。
2)将待测化合物用二甲基亚砜(DMSO)配置成10mM的储存液。临用前再用细胞培养液稀释,取20μl稀释好的化合物加到细胞培养孔,最终8个浓度梯度(300,100,10,3,1,0.3,0.1,0.03nM),每个浓度做1个孔,轻轻振荡混匀。另外设置阴性对照孔(只加培养液)和阳性对照孔(只加细胞和DMSO)。
3)细胞在孵育箱中培养16小时后,移除培养基并用冰冷的1X PBS润洗细胞一次。
4)弃掉PBS,细胞培养板每孔添加50ul冰冷的1X细胞裂解缓冲液,放在冰上孵育15分钟后震荡混匀。
5)向新的96孔板加入115ul样品稀释剂,从细胞培养板吸取5ul细胞裂解液上清加入到样品稀释液中,震荡均匀。
6)吸取100ul稀释过的样品到ELISA板中,用塑料胶膜密封ELISA板后放在37℃孵箱中孵育2小时。
7)轻轻取下胶膜,加1X洗涤缓冲液每孔200μl,震荡5min,倒掉,拍干。洗涤4次。
8)向ELISA板每孔添加100μl新配制的检测抗体(绿色)。用塑料胶膜密封后放在37℃孵箱中孵育1小时。
9)轻轻取下胶膜,倒掉检测抗体,拍干,加1X洗涤缓冲液每孔200μl,震荡5min,倒掉,拍干。洗涤4次。
10)向ELISA板每孔添加100μl新配制的HRP标记二抗(红色)。用塑料胶膜密封后放在37℃孵箱中孵育30分钟。
11)轻轻取下胶膜,倒掉二抗,拍干,加1X洗涤缓冲液每孔200μl,震荡5min,倒掉,拍干。洗涤4次。
12)向ELISA板每孔添加100μl TMB底物,放在37℃孵箱中孵育5分钟。
13)向ELISA板每孔添加100μl STOP溶液,温和振摇数秒。
3.结果读取
用无绒薄布擦拭各孔底部。添加STOP溶液后30分钟内,在450nm处读取吸光度。利用公式:剩余AR%=100*(检测孔OD值-空白对照OD值)/(阳性对照孔OD值-空白对照孔OD值),数据用软件GraphPad Prism5中Dose-response方程分析,得出DC50,Dmax值。DC50是降解50%目标蛋白时对应的化合物浓度。Dmax是目标蛋白最大程度能降解的百分比。
实验结果:
本发明化合物在100nM浓度下对雄激素受体(AR)的降解活性如表2所示,其中,%降解为降解的AR的百分比。本发明化合物在浓度梯度下对雄激素受体(AR)的降解活性如表3-1、3-2所示。表3-1、3-2中,DC50为将雄激素受体降解50%时的化合物浓度。Dmax为最高能降解的雄激素受体(AR)百分比。
可以看出,本发明化合物能够有效降解雄激素受体,大部分化合物对雄激素受体的DC50在0.3μM以下,有的甚至<10nM;大部分化合物对雄激素受体的Dmax在40%以上,有的甚至>80%。
表2.100nM浓度下化合物对雄激素受体(AR)降解活性
A:>80%,B:79%-50%,C:49%-25%,D:<25%
表3-1.化合物对雄激素受体(AR)降解的DC50和Dmax
DC50:A:<10nM,B:10-100nM,C:0.1-0.3μM,D:>0.3μM
Dmax:++++:>80%,+++:80%-60%,++:60%-40%,+:<40%
表3-2化合物对雄激素受体(AR)降解的DC50和Dmax
DC50:A:<10nM,B:10-100nM,C:0.1-0.3μM,D:>0.3μM
Dmax:++++:>80%,+++:80%-60%,++:60%-40%,+:<40%
实验例3蛋白免疫印迹实验(Western Blot)测定化合物物对雄激素受体(AR)蛋白的降解活性
实验材料:
前列腺癌VCaP细胞(ATCC,CRL-2876)
FBS(Gibco,Cat.No.10099-141)
0.01M PBS(Biosharp,Cat.No.162262)
Metribolone(R1881)(Macklin Biochemical,CAS.No.965-93-5)
DMEM/HIGH GLUCOSE(Hyclone,Cat.No.SH30243.01)
Penicillin-Streptomycin(Hyclone,Cat.No.SV30010)
二甲亚砜DMSO(Sigma,Cat.No.D5879)
离心试管,15ml(Excell Bio,Cat.No.CS015-0001)
细胞培养盘,(Excell Bio,Cat.No.CS016-0128)
6-well cell culture cluster(Corning,Cat.No.3516)
缓冲液RIPA lysate buffer(Beyotime,Cat.No.P0013B)
缓冲液Protein Loding Buffer(Beyotime,Cat.No.P0015L)
BCA蛋白检测盒BCA Protein Assay Kit(Beyotime,Cat.No.P0012)
SDS-PAGE凝胶制备试剂盒(成都佰和科技有限公司,Cat.No.PG112)
Anti-β-Actin rabbit mAb(CST,Cat.No.4970)
雄激素受体Androgen Receptor(D6F11)XP Rabbit mAb(CST,Cat.No.5153)
Peroxidase Affinipure(HRP)Goat Anti-Rabbit IgG(Zen Bioscience,Cat.No.511203)
TBST(Biosharp,Cat.No.BL601A)
ECL化学发光试剂盒(Beyotime,Cat.No.P0018)
实验方法:
1.缓冲液配制
2.实验步骤:
1)VCaP细胞用细胞培养液传代培养后,取生长状态良好的细胞接种于12孔板,每孔1ml,每孔细胞数为50万,于37℃,5%CO2细胞孵育箱中培养过夜。
2)将待测化合物用二甲基亚砜(DMSO)配置成10mM的储存液。临用前再用DMSO稀释3倍,取2μl稀释好的化合物加到细胞培养孔(以此保证培养体系中DMSO浓度为0.1%),每个浓度做2个孔重复,轻轻振荡混匀。另外设置阴性对照孔(加等量DMSO)和阳性对照孔。
3)培养16小时后,用RIPA细胞裂解液裂解细胞,提取蛋白,用BCA试剂盒测蛋白浓度。加5倍浓缩的蛋白上样缓冲液,100℃加热5分钟后样品放-20℃保存。
4)每孔蛋白量为30μg的蛋白量上样到聚丙烯酰胺凝胶,进行电泳。
5)蛋白质从聚丙烯酰胺凝胶转移到PVDF膜上,加5%脱脂牛奶室温封闭1小时,一抗(Androgen Receptor(D6F11)XP Rabbit mAb和Anti-β-Actin Rabbit mAb)4℃孵育过夜,TBST溶液洗膜三次每次10分钟,二抗(辣根过氧化物酶标记羊抗小鼠IgG)室温孵育2小时,再用TBST溶液洗膜三次每次10分钟。
3.结果检测:
最后加ECL显色液显色,用自动化学发光仪拍照,收集图片并分析。
实验结果:
用蛋白免疫印迹实验(Western Blot)方法测得本发明化合物在100nM浓度下对雄激素受体(AR)的降解活性如表4所示,%降解为降解的AR的百分比。可以看出,本发明化合物能够有效降解雄激素受体。
表4.本发明化合物对AR的降解活性
A:>80%,B:79%-50%,C:49%-25%,D:<25%
图1为化合物99在不同浓度下的蛋白免疫印迹实验结果,从照片中可以看出,本发明化合物对AR的降解活性是浓度依赖性的,降解活性随化合物浓度的增加而升高。
实验例4本发明化合物的代谢稳定性实验
1、材料仪器
液相系统(Shimadzu)、质谱系统(API 4000 instrument from AB Inc(Canada)with an ESI interface)、色谱柱(ACE Excel 3 AQ 30×2.1mm Column)、人肝药酶(Corning,Cat.#452117)、磷酸盐缓冲液、超纯水、MgCl2溶液、NADPH。
2、方法及结果
10μl 20mg/ml的肝微粒体和40μl的10mM NADPH一起加到孵育管中。肝微粒体和NADPH的终浓度分别为0.5mg/mL和1mM。同时准备一组不加NADPH加等量的超纯水做对照组。然后加入4μl浓度为200μM的对照化合物(维拉帕米)或待测化合物。化合物的终浓度为2μM。分别于孵育0,15,30,45和60min时取出50μl反应液同时加4倍体积的冰乙腈终止反应。取出的样品离心40min(3220g)后取出100μl上清液,往清液中加100μl超纯水混匀用于LC-MS/MS检测。最后计算肝微粒体代谢稳定性实验参数。
实验结果显示本发明的化合物显示了良好的代谢稳定性。
实验例5本发明化合物的药代动力学
1、实验材料及仪器:
LC-20AD高效液相色谱系统(日本SHIMADZU(岛津)公司)
API4000三重四极杆质谱仪,(美国Applied Biosystem公司)
PhenixWinnolin药动学软件(Version 6.3,美国Certara公司)
高速冷冻离心机(Thermo Fisher Scientific)
分析天平(赛多利斯,SECURA225D-1CN)
实验动物:ICR小鼠(成都达硕实验动物有限公司)
DMSO(Sigma)
CMC-Na(成都科龙化工)
肝素(成都科龙化工)
2、实验方法及结果
按给药剂量精密称取待测化合物,加入溶媒至终体积10ml,超声涡旋混匀。配置成浓度为0.5mg/ml的溶液。取配制的终溶液0.2ml,于-20℃保存,用于浓度测定。健康成年ICR小鼠9只(20-30g),禁食过夜(自由饮水)后,灌胃给药,给药体积0.2ml/10g;于给药前及给药后0.5,1,2,4,6,8,12,24h由眼眶后静脉丛采血0.1ml,4℃离心5min分离血浆,于-20℃保存待测。然后采用LC/MS/MS法测定血浆中的待测化合物浓度。
实验结果显示本发明的化合物显示了良好的药代动力学。
综上,本发明提供了式(I)所示的一类化合物,该类化合物能够靶向降解前列腺癌细胞中的雄激素受体,并且抑制前列腺癌细胞的增殖,同时还显示了良好的代谢稳定性和药代动力学性质。本发明化合物在制备雄激素受体的蛋白降解靶向嵌合体,以及制备治疗受雄激素受体调控的相关疾病(包括前列腺癌、乳腺癌、肯尼迪氏病)的药物中具有良好的应用前景。