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CN117658987A - Compounds as PLK1 inhibitors, and preparation method and application thereof - Google Patents

Compounds as PLK1 inhibitors, and preparation method and application thereof Download PDF

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Publication number
CN117658987A
CN117658987A CN202311140853.0A CN202311140853A CN117658987A CN 117658987 A CN117658987 A CN 117658987A CN 202311140853 A CN202311140853 A CN 202311140853A CN 117658987 A CN117658987 A CN 117658987A
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independently selected
mmol
alkyl
alkoxy
alkenyl
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李冬冬
张晓敏
朱正诞
王冬冬
孙伟杰
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Shanghai Shenshi Weisi Technology Co ltd
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Shanghai Shenshi Weisi Technology Co ltd
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Abstract

The present invention provides a class of compounds as PLK1 inhibitors, which are capable of inhibiting PLK1 and thus useful in the prevention and/or treatment of PLK 1-related diseases, such as tumors, and the like, and compositions comprising the same.

Description

Compounds as PLK1 inhibitors, and preparation method and application thereof
Technical Field
The invention provides a compound serving as a PLK1 inhibitor, and a preparation method and application thereof. The compounds and compositions comprising the same are useful for the prevention and/or treatment of tumor-related diseases.
Background
PLK1 belongs to the Polo-like kinase (PLKs) family (including PLK 1/2/3/4), and is a highly conserved Ser/Thr protein kinase, which plays an important role in the normal operation of cell cycle, centrosome maturation, cytoplasmic separation and the like. PLK1 regulates G2/M conversion by phosphorylating downstream cyclin such as WEE1/CDK1 and the like; PLK1 inhibition resulted in cell mitosis arrest in the G2 phase and induction of apoptosis (Sci Signal.2018Aug 14;11 (543): eaar 4195). In addition, PLK1 also has a regulatory effect on cell functions other than mitosis, such as PLK1 repair of damaged DNA, proliferation of tumor cells (Mol cell.2021Mar4; 81 (5): 1084-1099.e6.). PLK1 overexpression in most malignant tumors including colorectal, breast, lung, prostate, leukemia, pancreatic, etc., mediates poor patient prognosis (Genes 2019,10,208;World J Gastroenterol 2005;11 (36): 5644-5650). In addition, PLK1 is highly associated with multiple targets and pathways associated with tumors, RAS mutations cause cellular mitotic stress, making tumor cells more dependent on PLK1 for mitosis, interfering with the mitosis process can promote synthetic death of tumor cells, PLK1 knockdown or inhibit PLK1 kinase activity, making RAS mutant cells mitotically arrested in G2/M phase (cell 137,835-848, may 29, 2009); PLK1 can directly phosphorylate p53 protein to mediate unstable p53, or indirectly promote TOPORS and GTSE1 phosphorylation, degrade p53 and lose cancer inhibiting effect; PLK1 directly stabilizes or promotes the expression of oncogenic protein Myc and promotes the occurrence and development of tumors; PLK1 is also able to inhibit PTEN phosphorylation, activating PI3K signaling pathways.
Therefore, the enzyme activity of the small molecule targeted PLK1 has wide therapeutic effect on PLK1 related tumors.
Disclosure of Invention
The present invention provides a novel PLK1 inhibitor which is useful for the prevention and/or treatment of PLK 1-related diseases, such as tumors.
According to one aspect of the present invention, there is provided a compound of formula (I):
wherein,
e is independently selected from C, N, O;
t, M, Q is independently selected from N, CR3;
x, Y, Z is independently selected from N, CR;
ring a is selected from phenyl, containing 1-3 5-11 membered heteroaryl groups independently selected from N, O, S;
the B ring is connected with the ring carbon atom or the ring nitrogen atom of the A ring;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, optionally one or more of which are independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroarylOptionally one or more independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from one or more of F, cl, br, OH, CN, -NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S;
r4 is selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, alkyl, alkenyl,Alkynyl, alkoxy, cycloalkyl are optionally one or more independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r6 is selected fromNR8R9、NR8C(O)R9、C(O)NR8R9、C 1-4 Alkyl, piperazinyl, bridged piperazinyl, spiropiperazinyl, said alkyl, piperazinyl, bridged piperazinyl, spiropiperazinyl optionally substituted with one or more R7;
Or R6 taken together with the ring C atom to which it is attached, the ring Y atom, and the bond between the ring C atom and the ring Y atom form a 5-7 membered heterocycloalkyl containing 1-3 members independently selected from N, O, S, the ring optionally being one or more members independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r7 is independently selected from H, F, cl, br, OH, CN, NR R9, = O, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, said alkyl, alkoxy, cycloalkyl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, said alkyl, alkenyl, alkynyl, alkoxy optionally being substituted with one or more groups selected from F, cl, br, OH, CN, NH 2 Is substituted by a group of (2);
when R8 and R9 are attached to the same atom, R8 and R9 together with the atom to which they are attached may form a chain containing 1 to 3 groups independentlyA 3-7 membered heterocycloalkyl selected from N, O, S, optionally substituted with one or more substituents independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
m is 0, 1, 2 or 3.
w is 0, 1, 2.
According to one embodiment of the present invention, there is provided a compound represented by the following formula (IA):
wherein,
t, M, Q is independently selected from N, CR3;
x, Y, Z is independently selected from N, CR;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, which cycloalkyl group may optionally be independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The alkoxy groups being optionally substituted by one or more of alkyl, alkenyl, alkynyl, alkoxy groups Each independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from one or more of F, cl, br, OH, CN, -NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl containing 1-3 groups independently selected from N, O, S3-7 membered heterocycloalkyl containing 1-3 5-6 membered heteroaryl groups independently selected from N, O, S;
r4 is selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r7 is independently selected from H, F, cl, br, OH, CN, NR R9, = O, C 1-4 Alkyl, C 3-7 Cycloalkyl groups, said alkyl and cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, said alkyl, alkenyl, alkynyl, alkoxy optionally being substituted with one or more groups selected from F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
when R8 and R9 are attached to the same atom, R8 and R9 together with the atom to which they are attached may form a 3-7 membered heterocycloalkyl containing 1-3 members independently selected from N, O, S, optionally substituted with one or more members independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groupOptionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
m is 0, 1, 2 or 3;
n is 0, 1, 2, 3, 4, 5, 6, 7 or 8.
In a preferred embodiment:
t is N, M, Q are independently selected from CR3;
x, Y, Z is independently selected from CR5;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 An alkyl group, optionally one or more independently selected from F, cl, br, OH, CN, NH 2 Is substituted by a group of (2);
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, which cycloalkyl group may optionally be independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups are substituted;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl radicalsContaining 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being selected independently from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups are substituted;
r4 is selected from C 1-4 Alkoxy optionally substituted with one or more groups independently selected from F, cl, br;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 An alkyl group, optionally one or more independently selected from F, cl, br, OH, CN, NH 2 Is substituted by a group of (2);
r7 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 3-7 Cycloalkyl groups, said alkyl and cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy groups, said alkyl groups, alkoxy groups optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
m is 0, 1, 2 or 3;
n is 0, 1, 2, 3 or 4.
According to one embodiment of the present invention, there is provided a compound represented by the following formula (IC):
wherein,
x, Y is independently selected from N, CR;
r1 is independently selected from H, OH, = O, C 1-4 An alkyl group;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r2 is independently selected from H, C 1-4 Alkyl, -S (O) 2 R8、-P (O) R8R9, said alkyl groups optionally being independently selected from one or more of OH, C 1-4 The group of the alkoxy group is substituted;
r3 is selected from H, F, cl, br, CN, C 1-4 An alkyl group;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, cyclopropenyl optionally substituted with one or more groups independently selected from F, cl, br, CN, OH;
r5 is independently selected from H, CN;
r7 is independently selected from H, C 1-4 An alkyl group optionally substituted with one or more groups independently selected from F, cl, br, OH, CN;
r8 and R9 are independently selected from NH 2 、C 1-4 An alkyl group.
According to one embodiment of the present invention, there is provided a compound represented by the following formula (ID):
wherein,
r1 is independently selected from H, =o, methyl;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r3 is selected from H, F, cl, br, CN and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, cyclopropenyl optionally substituted with one or more groups independently selected from F, cl, br, CN, OH.
According to one embodiment of the present invention, there is provided a compound of formula (IB) or a pharmaceutically acceptable salt, deuterate thereof:
wherein,
r1 is independently selected from H, methyl;
r3 is selected from H, F, cl, br and methyl.
According to one embodiment of the present invention, there is provided a compound represented by the following formula (IE):
wherein,
r1 is independently selected from H, methyl;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r3 is selected from H, F, cl, br and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, said alkyl, alkoxy optionally substituted with one or more groups independently selected from F, cl, br.
According to one embodiment of the present invention, there is provided a compound of the following formula (IF):
wherein,
r3 is selected from H, F, cl, br and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, said alkyl, alkoxy optionally substituted with one or more groups independently selected from F, cl, br; preferably methoxy, trifluoromethoxy;
r6 is selected from
In a preferred embodiment, the compound of the invention is selected from the following compounds or pharmaceutically acceptable salts, deuterides thereof:
in the present invention, although substituents are disclosed as groups or ranges, the groups or ranges of the present invention are specifically intended to refer to each specific group that they cover. For example, the term "C 1-4 Alkyl "particularly refers to an independently disclosed methyl group (i.e., C 1 Alkyl), ethyl (i.e. C 2 Alkyl), propyl (i.e. C 3 Alkyl), butyl (i.e. C 4 Alkyl).
In the present invention, examples of "5-7 membered heterocycloalkyl groups containing 1 to 3 groups independently selected from N, O, S" include, but are not limited to, tetrahydrofuran, thiolane, pyrrolidine, pyrrolidone, pyrroline, dioxolane, oxazolidine, oxazolidone, oxazoline, isoxazolidine, thiazolidine, isothiazolidine, thiazoline, imidazolidine, imidazoline, pyrazolidine, pyrazoline, tetrahydropyran, dihydropyran, pyran, piperidine, piperidone, 1, 4-dioxane, morpholinone, piperazine, azepane, epoxyhexane, thietane, 1, 4-oxazaidine, 1, 4-thiazaidine.
In the present invention, examples of "containing 1 to 3-7 membered heterocycloalkyl groups independently selected from N, O, S" include, in addition to the above, ethylene oxide, aziridine, oxetane, N-oxetane, α -lactam ring, β -lactone.
In the present invention, examples of "containing 1 to 3 5-6 membered heteroaryl groups independently selected from N, O, S" include, but are not limited to, furan, thiophene, pyrrole, oxazole, isoxazole, thiazole, isothiazole, pyrazole, imidazole, 1,2, 3-triazole, 1,2, 4-triazole, oxadiazole, thiadiazole, pyridine, pyridone, pyrimidine, pyridazine, pyrazine, triazine.
In the present invention, examples of "containing 1 to 3 5-11 membered heteroaryl groups independently selected from N, O, S" are in addition to the above, also included, but not limited to, are benzoazetidines, benzobeta-lactam rings, benzobeta-lactones, benzooxazoles, benzofurans, benzothiophenes, benzopyrroles, benzopyrazoles, benzimidazoles, benzothiazoles, benzisothiazoles, benzoxazoles, benzisoxazoles, benzoisoxazoles, benzothiophenes, benzotriazoles, and the like benzotriazole, benzoxadiazole, benzothiadiazole, benzodioxan, benzomorpholine, benzopiperidine, benzopyran, benzopyridine, benzopyrimidine, benzotriazine, benzopiperidone, benzomorpholone, benzoazepine, benzopyran, benzopyrimidine, benzotriazine, benzopiperidone, benzomorpholone, benzotriazol pyridoazetidines, pyridobeta-lactam rings, pyridobeta-lactones, pyridooxadienes, pyridofurans, pyridothiophenes, pyridopyrroles, pyridopyrazoles, pyridoimidazoles, pyridothiazoles, pyridoisothiazoles, pyridooxazoles, pyridoisoxazoles, pyridotriazoles, pyridooxadiazoles, pyridothiadiazoles, pyridodioxanes, pyridomorpholines, pyridopiperidines, pyridopyrans, pyridopyridinepyridines, pyridopyrimidines, pyridotriazines, pyridopiperidones, pyridomorpholinones, pyridoazepinones.
The compounds of the invention may be asymmetric, e.g., have one or more stereocenters. All stereoisomers, such as enantiomers and diastereomers, are included within the scope of the invention unless otherwise indicated. In the present invention, the compound containing an asymmetrically substituted carbon atom may be isolated in any of optically active forms or racemic forms. Various methods for preparing optically active forms are known in the art, for example by resolution of the racemic mixture or by stereospecific synthesis.
The invention also includes pharmaceutically acceptable salts of the compounds. The compounds of the present invention can be used to prepare pharmaceutically acceptable salts by reaction with non-toxic inorganic or organic acids. Inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, nitric acid, hydrogen sulfate, boric acid, semi-sulfuric acid, and the like; organic acids such as formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, enanthic acid, undecanoic acid, palmitic acid, stearic acid, oleic acid, oxalic acid, malonic acid, adipic acid, lactic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, ascorbic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, benzoic acid, camphoric acid, citric acid, fumaric acid, gluconic acid, and various amino acids, and the like.
The invention also includes hydrates and solvates of the compounds.
The invention also includes all forms of the compounds whose atoms are in various isotopes. Isotopes include all atoms having the same atomic number but different mass numbers. Isotopes of hydrogen include deuterium, for example.
The invention also includes prodrugs of the compounds. "prodrug" refers to a compound that is structurally modified to have no or little activity in vitro but is released in vivo by enzymatic or non-enzymatic conversion to exert its potency.
Those skilled in the art will appreciate that the compounds of the present invention may be prepared by various methods disclosed in the literature. The compounds of the present invention may be prepared by reaction in a suitable solvent, which may be readily selected by those skilled in the art of organic synthesis to be substantially unreactive with the reactants, intermediates or products. The reaction may be carried out in one solvent or in a mixture of more than one solvent. The compounds of the present invention may be prepared by reaction at a suitable temperature, for example, between the solidification temperature of the solvent and the boiling temperature of the solvent. The method of preparing the compounds of the present invention involves protecting and deprotecting various chemical groups, and one skilled in the art of organic synthesis can readily determine whether protection and deprotection of a chemical group is required and select an appropriate protecting group. Any method known in the art may be used to monitor the reaction for preparing the compounds of the present invention, such as nuclear magnetic resonance spectroscopy, infrared spectroscopy, mass spectrometry, chromatography, and the like.
The compounds of the invention can be prepared, for example, by the following methods:
method I:
the groups in the formulae are as defined above.
Wherein the synthesis reaction of I-b may be performed in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from tetrahydrofuran, 1, 4-dioxane, tertiary butanol, toluene, xylene, DME, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base can be selected from lithium bis (trimethylsilyl) amide, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, etc.; the ligand may be selected from Davephos, xantphos and the like; the catalyst may be selected from Pd 2 (dba) 3 、Pd(OAc) 2 Etc.
Wherein the synthesis of I-c may be carried out in a solvent in the presence of a base. The solvent may be selected from tetrahydrofuran, 1, 4-dioxane, toluene, xylene, DME, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base may be selected from lithium bis (trimethylsilyl) amide, lithium diisopropylamide, potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, etc.
Wherein the synthesis reaction of I-d can be carried out in a solvent in the presence of a reagent. The solvent can be selected from acetonitrile, tetrahydrofuran, toluene, methanol, 1, 4-dioxane, etc. or any mixture thereof; the reactant can be selected from trimethyl iodosilane, trimethyl bromosilane, trimethyl chlorosilane, etc.;
Wherein the synthesis of I-e can be carried out in a solvent in the presence of a reagent. The reactant can be selected from phosphorus oxychloride, thionyl chloride and the like; the solvent may be selected from toluene, xylene, 1, 4-dioxane, methylene chloride, etc. or any mixture thereof.
Wherein the synthesis of compound (I) can be carried out in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from 1, 4-dioxane, tetrahydrofuran, water, methanol, toluene, xylene, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, sodium bicarbonate, etc.; the ligand may be selected from triphenylphosphine, dppp, dppf, etc.; the catalyst is selected from palladium acetate, palladium tetraphenyl phosphine and Pd 2 (dba) 3 、Pd(dppf)Cl 2 、Pd(PPh 3 ) 2 Cl 2 Etc.
Method II:
the groups in the formulae are as defined above.
Wherein the synthesis reaction of II-b can be carried out in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from 1, 4-dioxane, tetrahydrofuran, water, methanol, toluene, xylene, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, sodium bicarbonate, etc.; the ligand may be selected from triphenylphosphine, dppp, dppf, etc.; the catalyst is selected from palladium acetate, palladium tetraphenyl phosphine and Pd 2 (dba) 3 、Pd(dppf)Cl 2 、Pd(PPh 3 ) 2 Cl 2 Etc.
Wherein the synthesis of compound I can be carried out in the presence of a base, a ligand and a catalystIn the presence of a acylating agent in a solvent. The solvent is selected from tetrahydrofuran, 1, 4-dioxane, tertiary butanol, toluene, xylene, DME, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from potassium carbonate, sodium carbonate, cesium carbonate, lithium bis (trimethylsilyl) amide, potassium tert-butoxide, sodium tert-butoxide, etc.; the ligand may be selected from BINAP, davephos, xantphos and the like; the catalyst may be selected from Pd 2 (dba) 3 、Pd(OAc) 2 Etc.
Method III:
the groups in the formulae are as defined above.
Wherein the synthesis reaction of III-c can be carried out in a solvent in the presence of a reagent. The solvent may be selected from ethanol, methanol, water, etc. or any mixture thereof; the reactant may be a combination of cyanamide and hydrochloric acid, or a combination of S-methyl isothiourea and potassium carbonate.
Wherein the synthesis reaction of III-f can be carried out in a solvent in the presence of a reagent. The solvent can be selected from tetrahydrofuran, 1, 4-dioxane, methanol, water, etc. or any mixture thereof; the reactant may be selected from hydrochloric acid, sulfuric acid, and the like.
Wherein the synthesis of III-g can be carried out in a solvent in the presence of a reagent. The solvent may be selected from N, N-dimethylformamide, toluene, xylene, etc., or any mixture thereof; the reactant may be selected from N, N-dimethylformamide dimethyl acetal and the like.
Wherein the synthesis of compound I may be carried out in a solvent in the presence of a base. The solvent is selected from isopropanol, methanol, ethanol, tert-butanol, N-dimethylformamide, dimethyl sulfoxide, etc. or any mixture thereof; the base may be selected from potassium carbonate, sodium carbonate, potassium tert-butoxide, sodium hydroxide, potassium acetate, sodium acetate, etc.
Method IV:
the groups in the formulae are as defined above.
Wherein the synthesis of IV-b may be carried out in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from 1, 4-dioxane, tetrahydrofuran, water, methanol, toluene, xylene, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, sodium bicarbonate, etc.; the ligand may be selected from triphenylphosphine, dppp, dppf, etc.; the catalyst is selected from palladium acetate, palladium tetraphenyl phosphine and Pd 2 (dba) 3 、Pd(dppf)Cl 2 、Pd(PPh 3 ) 2 Cl 2 Etc.
Wherein the synthesis of IV-c may be carried out in a solvent in the presence of a reagent. The solvent is selected from dichloromethane, tetrahydrofuran, water, 1, 4-dioxane, methanol, toluene, xylene, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the reactant may be selected from Oxone, m-CPBA, etc.
Wherein the synthesis of compound I may be carried out in a solvent in the presence of a base. The solvent is selected from N, N-dimethylacetamide, N-dimethylformamide, dimethyl sulfoxide, toluene, xylene, etc. or any mixture thereof; the base may be selected from potassium tert-butoxide, sodium tert-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium acetate, sodium acetate, etc.
Method V:
the groups in the formulae are as defined above.
Wherein the synthesis of V-a can be carried out in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from tetrahydrofuran, 1, 4-dioxane, tertiary butanol, toluene, xylene, DME, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from potassium carbonate, sodium carbonate, cesium carbonate, lithium bis (trimethylsilyl) amide, potassium tert-butoxide, sodium tert-butoxide, etc.;the ligand may be selected from BINAP, davephos, xantphos and the like; the catalyst may be selected from Pd 2 (dba) 3 、Pd(OAc) 2 Etc.
Wherein the synthesis of compound I may be carried out in a solvent in the presence of a base, a ligand and a catalyst. The solvent is selected from tetrahydrofuran, 1, 4-dioxane, tertiary butanol, toluene, xylene, DME, dimethyl sulfoxide, N-dimethylformamide, etc. or any mixture thereof; the base is selected from lithium bis (trimethylsilyl) amide, potassium t-butoxide, sodium t-butoxide, potassium carbonate, sodium carbonate, cesium carbonate, potassium phosphate, etc.; the ligand may be selected from Davephos, xantphos and the like; the catalyst may be selected from Pd 2 (dba) 3 、Pd(OAc) 2 Etc.
The compounds of the invention are capable of inhibiting PLK1. Thus, according to another aspect of the invention, there is provided a method of inhibiting PLK1 using a compound of the invention.
According to another aspect of the present invention, there is provided a method for the prophylaxis and/or treatment of PLK 1-related disorders, wherein a therapeutically and/or prophylactically effective amount of a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention is administered to an individual in need thereof. PLK 1-associated diseases include any disease directly and/or indirectly associated with the expression and/or activity of PLK1, e.g., diseases associated with the overexpression of PLK 1; diseases and the like which are prevented and/or treated by modulating, for example, the inhibition of PLK1 activity.
PLK 1-related diseases include solid tumors, hematological tumors, and the like. Examples of solid tumors include, but are not limited to, colorectal cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, gastric cancer, head and neck cancer, ovarian cancer, uterine cancer, glioma, liver cancer, esophageal cancer, bladder cancer, renal cancer, lymphoma, melanoma, osteosarcoma, and the like; examples of hematological neoplasms include, but are not limited to, leukemia, myelodysplastic syndrome, and the like.
According to another aspect of the invention there is provided the use of a compound of the invention in the preparation of a PLK1 inhibitor.
According to another aspect of the invention, the invention provides the use of a compound of the invention in the manufacture of a medicament for the prophylaxis and/or treatment of PLK 1-related diseases.
When the compounds of the present invention are used for the prevention and/or treatment of PLK 1-related diseases, the compounds of the present invention may be administered in the form of a pharmaceutical composition. Thus, according to another aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier.
It will be appreciated by those skilled in the art that the pharmaceutical compositions of the present invention may be prepared by various methods disclosed in the literature. The compounds or pharmaceutical compositions of the present invention may be administered by a variety of routes, depending on the area in need of local or systemic treatment and the area in need of treatment. For example, administration may be by oral, parenteral (e.g., intravenous, arterial, subcutaneous, intraperitoneal, intramuscular injection or infusion), intracranial, e.g., intrathecal or intraventricular, transdermal, ocular, nasal, vaginal, rectal, pulmonary (e.g., by inhalation or insufflation of a powder or aerosol).
For administration by the oral route, the pharmaceutical compositions of the present invention are typically provided in the form of tablets, capsules or solutions. The tablets may contain a compound of the invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. Such carriers include, but are not limited to, diluents, disintegrants, binders, lubricants, colorants, or preservatives. The capsule comprises hard capsule and soft capsule. For parenteral administration, the pharmaceutical compositions of the present invention may be administered by intravenous injection, intramuscular injection or subcutaneous injection. It is usually provided as a sterile aqueous solution or suspension or lyophilized powder and is adjusted for proper pH and isotonicity.
The effective amount of the compounds of the present invention may be determined according to the particular use of the treatment, the mode of administration, the condition of the individual in need thereof, e.g., a patient. It is within the ability of one skilled in the art to determine an effective amount of a compound of the present invention. Typical dosage ranges are, for example, 1. Mu.g/kg/day to 1000 mg/kg/day.
When preventing and/or treating PLK 1-related diseases, the compounds of the present invention may be used in combination with one or more other drugs. Other drugs such as immune checkpoint drugs, targeted drugs, chemotherapeutic drugs, and the like.
Detailed Description
The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. Further, it is understood that various changes and modifications of the invention will become apparent to those skilled in the art upon reading the description herein, and such equivalents are intended to fall within the scope of the invention as defined by the appended claims.
Preparation example 1 Synthesis of common intermediates common int-1 and common int-2
Synthesis of 5- (4-methylpiperazin-1-yl) -2- [ (trifluoromethyl) oxy ] aniline (common int-1):
the compound 2-trifluoromethoxy-5-bromoaniline (20.0 g,78.1 mmol), tris (dibenzylideneacetone) dipalladium (Pd) 2 dba 3 To the reaction was added 1M solution of lithium bistrimethylsilylaminide in tetrahydrofuran (LiHMDS, 187mL,187.2 mmol) and N-methylpiperazine (12.5 g,124.8 mmol) under nitrogen protection, the reaction was heated to reflux overnight, and the solution was dissolved in tetrahydrofuran (50 mL) with 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (Davephos, 630mg,1.6 mmol). Cooled to room temperature, the crude product obtained by concentration was dissolved in methylene chloride (200 mL), the organic phase was washed with water (50 mL. Times.2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the concentrated crude product was purified by reverse phase Flash (acetonitrile: water 10% -70%, +0.1% ammonia) to give a pale yellow solid product (13.0 g). LC-MS: [ M+H ]] + :276.0. Synthesis of 4-methoxy-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (2):
the compound 5- (4-methylpiperazin-1-yl) -2- [ (trifluoromethyl) oxy]Aniline (common int-1,10g,36.33 mmol), 2-chloro-4-methoxypyrimidine (10.5 g,72.65 mmol) was dissolved in anhydrous tetrahydrofuran (50 mL), lithium bis (trimethylsilylamide) (72.65 mL,1 mol/L) was added dropwise under nitrogen protection at 0deg.C, and the reaction mixture was allowed to react for 2 hours at 25deg.C. The reaction solution was poured into an iced ammonium chloride solution (50 mL), followed by extraction with ethyl acetate (50 mL. Times.3), The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness and purified by column chromatography (methanol: dichloroethane=1:200-1:20) to give white solid 2 (10.7 g, 77.0%). LC-MS: [ M+H ]] + :384.0。
Synthesis of 2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-ol (3):
the compound 4-methoxy-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (160 mg,0.42 mmol), sodium iodide (125 mg,0.84 mmol) was dissolved in acetonitrile (6 mL), and trimethylchlorosilane (135.8 mg,1.25 mmol) was then added dropwise and reacted at 90℃for 2 hours. H is added into the reaction solution 2 O (5 mL), ethyl acetate extraction, the mixture was washed with water and saturated aqueous sodium chloride in this order, the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was subjected to column chromatography (methanol: dichloromethane=1:100) to give a white solid (80 mg). LC-MS [ M-H ]] + :368.0。
Synthesis of 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2):
compound 2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-ol (80 mg) was added to phosphorus oxychloride (4 mL) and reacted at 80 ℃ for 2 hours. The reaction solution was concentrated, and the crude product was suspended in ethyl acetate (10 mL), the pH was adjusted to about 9 with saturated sodium bicarbonate under ice bath, and the ethyl acetate layer was collected, dried, filtered and concentrated to give a white solid (30 mg) by column chromatography (methanol: dichloromethane=1:100). LC-MS: [ M+H ] ] + :388.0。
Preparation example 2 Synthesis of common intermediate common int-3
Synthesis of N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) carboxamide (common int-3):
a mixture of acetic anhydride (6.8 mL,72.65 mmol) and formic acid (4.1 mL,108.98 mmol) was stirred at room temperature for 30 minutes, and after the reaction temperature was lowered to 0deg.C, 5- & lt/EN & gt was slowly added dropwise4-methylpiperazin-1-yl) -2- [ (trifluoromethyl) oxy]A solution of aniline (common int-1,2.0g,7.27 mmol) in dichloromethane (3 mL) was stirred at room temperature for 2 hours. The solution was concentrated and diluted with ethyl acetate (20 mL), and washed with saturated sodium bicarbonate solution (20 mL) and water (20 mL), respectively. The organic layer was concentrated to give a colorless oil (2.1 g) which was used directly in the related reaction. LC-MS: [ M+H ]] + :304.0； 1 H NMR(400MHz,CDCl3).δ10.00(s,1H),8.32(d,J＝4.0,1H),7.84(d,J＝4.0Hz,1H),7.20(dd,J＝8.0,4.0Hz,1H),6.74(dd,J＝8.0,4.0Hz,1H),3.10-3.18(m,4H),2.43-2.45(m,4H),2.22(s,3H)。
Preparation example 3 Synthesis of common intermediate common int-5
Synthesis of 5-chloro-4-methoxy-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (1):
the compound 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (830.0 mg,3.02 mmol), 2, 5-dichloro-4-methoxypyrimidine (1.1 g,6.15 mmol) was dissolved in anhydrous tetrahydrofuran (10.0 mL) and lithium bis (trimethylsilylamide) (6.0 mL,1.0 mol/L) was added dropwise under nitrogen at 0deg.C and allowed to warm slowly to room temperature. The reaction was stirred at room temperature for 1 hour, quenched with 20mL of saturated ammonium chloride solution, then extracted with ethyl acetate (15 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column (dichloromethane: methanol=150:1-80:1) (dichloromethane: methanol=10:1, rf=0.3) to give a white solid (940 mg). LC-MS: [ M+H ] ] + :418.1,420.1。
Synthesis of 5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-ol (2):
the compound 5-chloro-4-methoxy-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (720.0 mg,1.72 mmol) was dissolved in acetonitrile (7.0 mL), trimethyliodosilane (1.0 g,5.00 mmol) was added dropwise, stirred at 80℃for 6 hours, saturated sodium bicarbonate solution was added to weakly alkaline, and extracted with ethyl acetate (15 mL×)3) The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane: methanol=25:1-10:1) (rf=0.1, dichloromethane: methanol=10:1) to give a white solid (400.0 mg). LC-MS: [ M+H ]] + :404.0,406.0。
Synthesis of 4, 5-dichloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-5):
the compound 5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-ol (320.0 mg,0.79 mmol) was dissolved in phosphorus oxychloride (2.0 mL) and stirred at 80 ℃ for 1 hour. The reaction solution was adjusted to weak base with saturated sodium bicarbonate solution, extracted with dichloromethane (15 ml×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified via column (dichloromethane: methanol=150:1-50:1) (rf=0.4, dichloromethane: methanol=10:1) to give a white solid (200 mg). 1 H NMR(400MHz,DMSO-d6)δ9.73(s,1H),8.55(s,1H),7.23–7.18(m,1H),7.14(d,J＝3.2Hz,1H),6.86–6.81(m,1H),3.18–3.10(m,4H),2.47–2.40(m,4H),2.22(s,3H)；LC-MS:[M+H] + :422.3,424.3。
Preparation example 4 Synthesis of common intermediate common int-6
Synthesis of 4-bromo-2-nitro-1- (2, 2-trifluoroethoxy) benzene (2):
the compound 4-bromo-2-nitrophenol (5.0 g,22.94 mmol) and cesium carbonate (15.0 g,46.04 mmol) were added to N, N-dimethylformamide (30 mL) and 2, 2-trifluoroethyl triflate (6.4 g,27.52 mmol) was slowly added dropwise under nitrogen at 0deg.C. The reaction was slowly warmed to room temperature and stirred for 3 hours. After dilution with 20mL of water, extraction with ethyl acetate (20 ml×3), drying the combined organic phases over anhydrous sodium sulfate, filtration, concentration and purification by column chromatography on silica gel (petroleum ether: ethyl acetate=20:1-10:1) (rf=0.4, petroleum ether: ethyl acetate=10:1) afforded a red solid (5.5 g). 1 H NMR(400MHz,CDCl 3 )δ7.96(d,J＝2.4Hz,1H),7.62(dd,J＝8.8,2.4Hz,1H),6.96(d,J＝9.2Hz,1H),4.40(m,2H)。
Synthesis of 1-methyl-4- (3-nitro-4- (2, 2-trifluoroethoxy) phenyl) piperazine (3):
the compound 4-bromo-2-nitro-1- (2, 2-trifluoroethoxy) benzene (1.9 g,6.33 mmol), N-methylpiperazine (1.0 g,9.98 mmol), tris (dibenzylideneacetone) dipalladium (576.9 mg,0.63 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (364.5 mg,0.63 mmol) and cesium carbonate (4.1 g,12.58 mmol) were added to anhydrous dioxane (20 mL) and reacted under nitrogen at 100℃for 16 hours. After dilution with water (10 mL), extraction with ethyl acetate (15 ml×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel (dichloromethane: methanol=50:1 to 20:1) (dichloromethane: methanol=20:1, rf=0.2) to give a white solid (1.1 g). LC/MS [ M+H ] ] + :320.1. Synthesis of 5- (4-methylpiperazin-1-yl) -2- (2, 2-trifluoroethoxy) aniline (common int-6):
the compound 1-methyl-4- (3-nitro-4- (2, 2-trifluoroethoxy) phenyl) piperazine (1.1 g,3.45 mmol) was dissolved in 10mL methanol and 50.0mg Pd/C was added. Stirring for 16 hours at room temperature under hydrogen atmosphere, filtering, concentrating, purifying by column (dichloromethane: methanol=20:1-10:1) (dichloromethane: methanol=10:1, rf=0.35) to give white solid (800.0 mg). 1 H NMR(400MHz,CDCl 3 )δ6.66(d,J＝8.8Hz,1H),6.30(d,J＝2.8Hz,1H),6.21(dd,J＝8.4,2.8Hz,1H),4.21(m,2H),3.72(s,2H),3.06–3.01(m,4H),2.52–2.46(m,4H),2.27(s,3H)；LCMS:[M+H] + :290.1。
Preparation example 5 Synthesis of common intermediate common int-7
Synthesis of 3, 3-dimethyl-6- (pinacol borate) isoindol-1-one (2):
to a suspension of 6-bromo-3, 3-dimethylisoindol-1-one (250.0 mg,1.04 mmol) and pinacol diboronate (396.1 mg,1.56 mmol) in dioxane (8 mL) was added [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane Complex (81.1 mg,0.10 mmol) and Potassium acetate [. Times.1 mg204.1mg,2.08 mmol). The reaction system was stirred at 80 ℃ for 4 hours under nitrogen protection and then concentrated directly to dryness. The crude product was purified by column on silica gel (ethyl acetate: petroleum ether=1:8, rf=0.2) to give a brown solid (280.0 mg). LC-MS: [ M+H ]] + :288.1。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-7):
To a stirred suspension of 3, 3-dimethyl-6- (pinacol borate) isoindol-1-one (100.0 mg,0.35 mmol) and 2, 4-dichloro-5-fluoropyrimidine (58.4 mg,0.35 mmol) in dioxane and water (3:1, 8 mL) under nitrogen was added [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (28.4 mg,0.035 mmol) and potassium carbonate (96.6 mg,0.70 mmol). Stirring at 70℃for 2 hours under nitrogen protection and concentrating directly to dryness. The crude product was purified by column on silica gel (ethyl acetate: petroleum ether=1:5, rf=0.3) to give the product as a yellow solid (100.0 mg). LC-MS: [ M+H ]] + :292.0,294.0。
Preparation example 6 Synthesis of common intermediate common int-8
Synthesis of N- (5-bromo-2- (trifluoromethoxy) phenyl) carboxamide (2):
a mixture of acetic anhydride (7.3 mL,78.12 mmol) and formic acid (4.4 mL,117.18 mmol) was stirred at room temperature for 30 min, then cooled to 0deg.C, a solution of 5-bromo-2- (trifluoromethoxy) aniline (2.0 g,7.81 mmol) in dichloromethane (3 mL) was slowly added and the mixture was stirred at room temperature for 2 h. The solution was concentrated to dryness and diluted with ethyl acetate (20 mL) and washed with saturated sodium bicarbonate solution (20 mL) and water (20 mL), respectively. The organic phase was dried over anhydrous sodium sulfate and concentrated to give crude product (2.1 g) as colorless oil. LC-MS: [ M+H ] ] + :283.9，285.9。
Synthesis of 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8):
6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-7) (446.3 mg, 1.5)3 mmol), N- (5-bromo-2- (trifluoromethoxy) phenyl) carboxamide (435.0 mg,1.53 mmol) and cesium carbonate (750.0 mg,2.30 mmol) were dissolved in acetonitrile (8 mL), reacted at 50℃for 18 hours, the reaction solution was dried by spin, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1) to give a white solid (540.0 mg). LC-MS: [ M+H ]] + :511.3,513.3。
Preparation example 7 Synthesis of common intermediate common int-9, common int-10
Synthesis of spiro [ cyclopropane-1, 1 '-isoindol ] -3' -one (2):
ethyl 2-cyanobenzoate (2.4 g,13.70 mmol) was dissolved in diethyl ether, tetraisopropyl titanate (4.3 g,15.13 mmol) was added dropwise to the reaction, followed by ethyl magnesium bromide in tetrahydrofuran (13.7 mL,2 mol/L). The reaction solution was stirred at room temperature for 16 hours. LCMS monitored reaction was complete. To the reaction solution was added a dilute hydrochloric acid solution (20 mL,1 mol/L), followed by extraction with methylene chloride (40 mL. Times.3). The organic phase was washed with saturated brine (30 mL), the organic phase was separated, dried, filtered, concentrated and then purified by column chromatography (methanol: dichloromethane=10%, rf=0.5) to give the product as a white solid (800.0 mg). LCMS [ M+H ] ] + :160.2。
Synthesis of 5' -bromospiro [ cyclopropane-1, 1' -isoindol ] -3' -one (3):
anhydrous aluminum trichloride (1.3 g,9.75 mmol) was placed in a three-necked flask and blanketed with nitrogen, followed by spiro [ cyclopropane-1, 1' -isoindole ]]3' -Ketone (600.0 mg,3.77 mmol) was dissolved in 1, 2-dichloroethane and then added followed by dropwise addition of bromine (783.0 mg,4.90 mmol). After the completion of the dropwise addition, the reaction mixture was heated to 80℃and stirred for 16 hours, and LCMS showed completion of the reaction. The reaction was quenched with saturated sodium thiosulfate (15 mL), extracted with ethyl acetate (3X 20 mL), the organic phases combined and washed once with saturated sodium chloride (15 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was subjected to column chromatography (ethyl acetate: petroleum ether=0-100%, ethyl acetate: petroleum ether=1:1, rf=0.5) to give a pale yellow solid (400.0 mg). LCMS [ M+H ]] + :238.1,240.1。
Synthesis of the pinacol 5' -borate spiro [ cyclopropane-1, 1' -isoindol ] -3' -one (common int-9):
5 '-bromospiro [ cyclopropane-1, 1' -isoindole]-3 '-one (400.0 mg,1.68 mmol), potassium acetate (495.0 mg,5.04 mmol), pinacol bisborate (512.0 mg,2.02 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (137.2 mg,0.17 mmol) was dissolved in 1, 4-dioxane (20 mL) and water (3 mL). The reaction was stirred at 100 ℃ for 16 hours under nitrogen, LCMS showed complete reaction. The reaction solution was poured into water (20 mL) and then extracted with ethyl acetate (15 ml×3), the organic phases were combined and washed once with saturated sodium chloride (15 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product as a brown solid (300.0 mg). The crude product was used directly in the next reaction. LCMS [ M+H ] ] + :286.3.5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindole]-synthesis of 3' -ketone (common int-10):
the compound 5 '-boronic acid pinacol ester spiro [ cyclopropane-1, 1' -isoindole ]]-3' -one (200.0 mg,0.70 mmol), cesium carbonate (455.0 mg,1.40 mmol), 1-bis (diphenylphosphine) ferrocene palladium dichloride dichloromethane complex (56.7 mg,0.07 mmol), 2, 4-dichloro-5-fluoropyrimidine (117.1 mg,0.70 mmol) were suspended in 1, 4-dioxane and water (10 ml, 5:1) and the reaction was reacted under nitrogen at 100 ℃ for 6 hours. The reaction mixture was cooled to room temperature, poured into water (20 mL), and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined and washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (ethyl acetate: petroleum ether=2:1, rf=0.6) to give a yellow solid (150.0 mg). LC-MS: [ M+H ]] + :290.2。
Preparation example 8 Synthesis of common intermediate common int-11
Synthesis of ethyl divinyl phosphonate (2):
the compound ethyl dichlorophosphate (10.0 g,61.39 mmol) was dissolved in dichloromethane (10 mL) and vinylmagnesium bromide (1M) was added at-65℃under nitrogenTetrahydrofuran solution) (123.5 mL,123.46 mmol), and the reaction was stirred at this temperature for 2h. Ethanol (20 mL) was slowly added to the reaction solution, and the mixture was gradually warmed to room temperature. The reaction solution was concentrated and purified by silica gel column chromatography (methanol: dichloromethane=0% -15%) to obtain a pale yellow liquid (3.2 g). LC-MS: [ M+H ] ] + :147.1。
Synthesis of 4-ethoxy-1-methyl-1, 4-azaphosphine-4-oxide (3):
after mixing ethyl divinyl phosphonate (2.0 g,13.70 mmol) with a solution of methylamine in THF (1 m,20 ml), the reaction was carried out for 20 hours at 70 ℃. The reaction solution was cooled and concentrated to obtain a crude product, and the crude product was purified by silica gel column chromatography (methanol: dichloromethane=0% -18%) to obtain a colorless liquid (1.0 g). LC-MS: [ M+H ]] + :178.1。
Synthesis of 1-methyl-1, 4-azaphosphine-4-oxide (common int-11):
the compound 4-ethoxy-1-methyl-1, 4-azaphosphine-4-oxide (1.0 g,5.65 mmol) was dissolved in tetrahydrofuran (10 mL), lithium aluminum hydride (214.4 mg,5.65 mmol) was slowly added at 0deg.C, and reacted at 0deg.C under nitrogen for 2 hours. To the reaction mixture was added water (0.2 mL), 15% aqueous sodium hydroxide solution (0.2 mL) and water (0.6 mL) in this order, and the mixture was stirred at room temperature for half an hour, and then the reaction mixture was filtered through celite, and the cake was washed with ethyl acetate. The filtrate was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by reverse phase column (acetonitrile: water=5% -40%) to give a white solid (200.0 mg). LC-MS: [ M+H ]] + :134.1。
Preparation example 9 Synthesis of common intermediate common int-12
Synthesis of 5' - (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (common int-12):
The compound N- (5-bromo-2- (trifluoromethoxy) phenyl) carboxamide (500 mg,1.76 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (463.6 mg,1.60 mmol) and cesium carbonate (1.0 g,3.07 mmol) were dissolved in acetonitrile (8 mL), displacedNitrogen was warmed to 80℃and stirred for 16 hours, LCMS monitored the starting material was reacted and the product was the main peak. The reaction solution was dried by spinning, water (20 mL) was added to dissolve, extraction was performed with ethyl acetate (20 ml×3), the combined organic phases were washed once with saturated brine (30 mL), dried over anhydrous sodium sulfate, and spun-dried to give a crude product, which was passed through a silica gel column (methanol: dichloromethane=0-2%, dichloromethane: methanol=10:1, rf=0.2) to give a pale yellow solid (260.0 mg). LC-MS: [ M+H ]] + :509.1,511.1。
Preparation example 10 Synthesis of common intermediate common int-13
Synthesis of 2-chloro-5- (difluoromethoxy) -4-iodopyridine (2):
to a solution of 6-chloro-4-iodopyridin-3-ol (4.7 g,18.40 mmol) in N, N-dimethylformamide (20 mL) was added cesium carbonate (7.79 g,23.92 mmol) and sodium 2-chloro-2, 2-difluoroacetate (5.61 g,36.80 mmol) in sequence. The mixture was stirred at 0℃for 3 hours. The reaction was diluted with ethyl acetate (200 mL), washed with water and brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=1:10) to give a white solid (4.4 g). LC-MS: [ M+H ] ] + :306.1,308.1。
Synthesis of N- (2-chloro-5- (difluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethane imine (3):
to a solution of 2-chloro-5- (difluoromethoxy) -4-iodopyridine (2.0 g,6.51 mmol) and benzophenone imine (1.4 g,7.86 mmol) in toluene (30 mL) were added sequentially 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (820.0 mg,1.31 mmol), tris (dibenzylideneacetone) dipalladium (600.0 mg,0.66 mmol) and cesium carbonate (4.3 g,13.19 mmol). The mixture was stirred at 60 ℃ for 1 hour until the starting material was completely consumed. The reaction mixture was cooled to room temperature and concentrated to dryness. The crude product was purified by column chromatography on silica gel (0-5% methanol/dichloromethane) to give a yellow oil (1.8 g). LC-MS: [ M+H ]] + :359.2,361.2。
Synthesis of N- (5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) -1, 1-diphenylmethanimine (4):
to a solution of N- (2-chloro-5- (difluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethane imine (400.0 mg,1.12 mmol) and 1-methylpiperazine (0.16 mL,1.45 mmol) in toluene (20 mL) were successively added 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (137.0 mg,0.22 mmol), tris (dibenzylideneacetone) dipalladium (102.1 mg,0.11 mmol) and sodium t-butoxide (214.3 mg,2.23 mmol), and the mixture was stirred at 110℃for 1 hour. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-5% methanol/dichloromethane) to give a yellow oil (430.0 mg). LC-MS: [ M+H ] ] + :423.2。
Synthesis of 5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-amine (common int-13):
to a solution of N- (5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) -1, 1-diphenylmethanimine (400.0 mg,0.95 mmol) in dichloromethane (2 mL) at room temperature was added trifluoroacetic acid (1 mL), and the reaction solution was stirred at room temperature for 3 hours and then concentrated to dryness. The resulting residue was suspended in hydrochloric acid (3 mL, 1N) and extracted with dichloromethane (10 mL. Times.2). The combined organic phases were washed again with hydrochloric acid (1N, 3 mL). The aqueous layers were combined and concentrated under reduced pressure to give a brown oil (120.0 mg). LC-MS: [ M+H ]] + :259.2。
Preparation example 11 Synthesis of common intermediate common int-14
Synthesis of tert-butyl 4- (3-bromo-4- (trifluoromethoxy) phenyl) -2- (hydroxymethyl) piperazine-1-carboxylate (2):
to a solution of 2-bromo-4-iodo-1- (trifluoromethoxy) benzene (4.8 g,13.08 mmol) in dimethyl sulfoxide (20 mL) was added tert-butyl 2- (hydroxymethyl) piperazine-1-carboxylate (3.4 g,15.72 mmol), cesium carbonate (5.12 g,15.71mmol, cuprous iodide (0.50 g,2.63 mmol) and L-proline (0.30 g,2.61 mmol), and the mixture was stirred under nitrogen at 60℃for 1 hour, after the reaction was completed, the reaction mixture was diluted with water (200 mL), extracted with ethyl acetate (200 mL. Times.3), the combined organic phases were washed with saturated aqueous sodium chloride (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give crude product, which was purified by silica Purification by column chromatography (petroleum ether: ethyl acetate=8:1) gave a yellow oily liquid (1.3 g). LC-MS: [ M+H ]] + :455.1,457.1。
Synthesis of (4- (3-bromo-4- (trifluoromethoxy) phenyl) piperazin-2-yl) methanol (3):
tert-butyl 4- (3-bromo-4- (trifluoromethoxy) phenyl) -2- (hydroxymethyl) piperazine-1-carboxylate (1.3 g,2.86 mmol) was dissolved in dichloromethane (8 mL), trifluoroacetic acid (4 mL) was added and stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was dried by spin-drying to give a pale yellow oil (960.0 mg). The crude product was used directly in the next reaction. LC-MS: [ M+H ]] + :355.1,357.1。
Synthesis of (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (common int-14):
(4- (3-bromo-4- (trifluoromethoxy) phenyl) piperazin-2-yl) methanol (390.0 mg,1.10 mmol) was dissolved in water (2 mL) and acetonitrile (5 mL), then 37% aqueous formaldehyde (0.5 mL) and acetic acid (20.0 mg,0.33 mmol) were slowly added, and sodium triacetoxyborohydride (694.9 mg,3.28 mmol) was added after stirring for 1 hour. After stirring the reaction solution at room temperature for 3 hours, it was quenched with saturated aqueous sodium bicarbonate (20 mL) and extracted with ethyl acetate (30 mL. Times.2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (280.0 mg). LC-MS: [ M+H ] ] + :369.1。
Preparation example 12 Synthesis of common intermediate common int-15
Synthesis of 5-bromo-3-iodo-2-methylbenzoic acid methyl ester (2):
methyl 2-methyl-3-amino-5-bromobenzoate (5.0 g,20.48 mmol) was dissolved in 6N hydrochloric acid (60 mL) at-5 ℃, sodium nitrite (1.6 g,23.19 mmol) was dissolved in water (20 mL) and added dropwise to the reaction system, sodium iodide (3.4 g,22.68 mmol) was dissolved in water (20 mL) after 30 minutes of reaction and added slowly dropwise to the reaction system, and the reaction solution was warmed to room temperature and at 90 DEG CThe reaction was carried out for 1 hour. The reaction was cooled, extracted with ethyl acetate (15 ml×3), the organic phase was washed with saturated aqueous sodium chloride (20 mL), dried and spun-dried, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1) to give a brown solid (4.8 g). 1 H NMR(400MHz,DMSO)δ8.25(d,J＝2.0Hz,1H),7.86(d,J＝2.0Hz,1H),3.84(s,3H),2.49(s,3H)。
Synthesis of methyl 5-bromo-2- (bromomethyl) -3-iodobenzoate (3):
methyl 5-bromo-3-iodo-2-methylbenzoate (4.8 g,13.38 mmol) was dissolved in carbon tetrachloride (70 mL), and then N-bromosuccinimide (2.4 g,13.48 mmol) and dibenzoyl peroxide (320.0 mg,1.32 mmol) were added sequentially, and the reaction was allowed to warm to 80℃for 18 hours, cooled to react, and the solvent was dried by spin-drying, and the crude product was subjected to column chromatography (ethyl acetate: petroleum ether=1:10) to give a yellow solid (5.5 g). 1 H NMR(400MHz,DMSO)δ8.38(d,J＝2.1Hz,1H),8.00(d,J＝2.1Hz,1H),4.97(s,2H),3.88(s,3H)。
Synthesis of 6-bromo-4-iodo-isoindol-1-one (4):
methyl 5-bromo-2- (bromomethyl) -3-iodobenzoate (5.5 g,12.68 mmol) was dissolved in 7M methanolic ammonia (60 mL) and the reaction stirred at room temperature for 2 hours. The reaction mixture was dried by spinning, diluted with saturated aqueous sodium hydrogencarbonate (20 mL), extracted with methylene chloride (20 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered and dried by spinning to give a yellow solid (4.2 g). LC-MS: [ M+H ]] + :338.1,340.1。
Synthesis of 6-bromo-4-iodo-1-oxoisoindole-2-carboxylic acid tert-butyl ester (5):
6-bromo-4-iodoisoindol-1-one (4.2 g,12.43 mmol), 4-dimethylaminopyridine (460.0 mg,3.77 mmol) and triethylamine (5.0 g,49.41 mmol) were dissolved in dichloromethane (100 mL), and di-tert-butyl dicarbonate (8.1 g,37.11 mmol) was slowly added dropwise to the reaction system and reacted at room temperature for 18 hours. The reaction system was diluted with dichloromethane (50 mL), washed with water (100 mL), and the organic phase was separated, dried, filtered and spin-dried. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate=5:1) to give a white solid (2.7 g): LC-MS: [ M ] t Bu+H] + :382.1,384.1。
Synthesis of 6-bromo-4-iodo-3-methyl-1-oxoisoindole-2-carboxylic acid tert-butyl ester (common int-15):
6-bromo-4-iodo-1-oxoisoindole-2-carboxylic acid tert-butyl ester (1.2 g,2.74 mmol) was dissolved in tetrahydrofuran (60 mL), 1M lithium bis trimethylsilylamide (3.3 mL,3.30 mmol) was added dropwise at-70℃and stirred at-70℃for half an hour. Methyl iodide (580.0 mg,4.09 mmol) was added dropwise to the reaction system, the reaction was warmed to room temperature and then stirred for 1 hour, the reaction was quenched with saturated aqueous ammonium chloride (20 mL), extracted with ethyl acetate (20 mL. Times.3), and the organic phase was washed with saturated aqueous sodium chloride (20 mL), dried and dried by spin-drying to give a white solid (1.1 g). LC-MS: [ M ] t Bu+H] + :396.0,398.0。
Preparation example 13 Synthesis of common intermediate common int-16
Synthesis of 1- (4-methoxy-3-nitrophenyl) -4-methylpiperazine (2):
the compound 4-bromo-1-methoxy-2-nitrobenzene (500.0 mg,2.16 mmol), 1-methylpiperazine (280.6 mg,2.80 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (88.5 mg,0.22 mmol), cesium carbonate (1404.2 mg,4.31 mmol) and tris (dibenzylideneacetone) dipalladium (98.7 mg,0.11 mmol) were dissolved in dioxane (8 mL), replaced with nitrogen three times, stirred for 6 hours at 90 ℃ and LCMS detected complete reaction. The reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-34%, petroleum ether/ethyl acetate=1:1, rf=0.2) to give a yellow oil (230.0 mg). LC-MS: [ M+H ]] + :252.1。
Synthesis of 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (common int-16):
the compound 1- (4-methoxy-3-nitrophenyl) -4-methylpiperazine (180.0 mg,0.72 mmol) and Pd/C (80 mg,10% w/w) were suspended in methanol (5 mL), and after three hydrogen substitutions, the reaction was stirred at room temperature for 4 hours. After completion of the reaction, LCMS was monitored, the reaction mixture was filtered through celite, and the cake was washed 3 times with ethyl acetate, filtered After the solution was dried by spinning, a yellow solid (150.0 mg) was obtained. LC-MS: [ M+H ]] + :222.2。
Preparation example 14 Synthesis of common intermediate common int-17
5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (600.0 mg,1.29 mmol), potassium carbonate (533.7 mg,3.86 mmol), ethylene potassium trifluoroborate (189.7 mg,1.42 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (47.1 mg,0.065 mmol) was dissolved in dioxane (6 mL) and water (1 mL), reacted at 95℃for 2 hours under nitrogen protection, the reaction solution was dried by spinning, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=5:1) to give a white solid (470.0 mg). LC-MS [ M-t-Bu+H ]] + :310.1,312.1。
Preparation example 15 Synthesis of common intermediate common int-18
Synthesis of 5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (common int-18):
6-bromo-4-iodo-3-methyl-1-oxoisoindole-2-carboxylic acid tert-butyl ester (1.1 g,2.43 mmol) was dissolved in tetrahydrofuran (60 mL), 1M lithium bis trimethylsilylamide (2.9 mL,2.90 mmol) was added dropwise with stirring at-70℃and stirring at-70℃for half an hour. After methyl iodide (1.0 g,7.05 mmol) was added dropwise to the reaction system, the reaction was warmed to room temperature and stirred for 0.5 hours, the reaction was quenched with saturated aqueous ammonium chloride (50 mL), extracted with ethyl acetate (50 ml×3), the organic phase was washed with saturated aqueous sodium chloride (100 mL), dried, and dried by spin-drying, and the crude product was subjected to column chromatography (petroleum ether: ethyl acetate=10:1) to give a white solid (180.0 mg). LC-MS: [ M ] t Bu+H] + :410.0,412.0。
Preparation example 16 Synthesis of common intermediate common int-19
Synthesis of 6-bromo-3, 3-dimethyl-4- (methylthio) isoindolin-1-one (common int-19):
the compound tert-butyl 5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylate (1.0 g,2.14 mmol), sodium methyl mercaptide (451.4 mg,6.44 mmol), cuprous iodide (40.9 mg,0.21 mmol), (L) -proline sodium salt (59.8 mg,0.43 mmol) was placed in dimethyl sulfoxide (10.0 mL). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (dichloromethane: methanol=20:1) to give a yellow solid (520.0 mg). LC-MS: [ M+H ]] + :286.0,287.9。
Preparation example 17 Synthesis of common intermediate common int-20
(E) -synthesis of 5-bromo-7- (2-ethoxyvinyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (1):
tert-butyl 5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylate (250.0 mg,0.54 mmol), (E) -2- (2-ethoxyvinyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan (130.0 mg,0.66 mmol), potassium phosphate (250.0 mg,1.18 mmol) and [1,1' -bis (diphenylphosphine) ferrocene ]Palladium dichloride (40.0 mg,0.05 mmol) was dissolved in dioxane (5 mL) and water (0.5 mL), and reacted at 100℃for 3 hours under nitrogen. The reaction system was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=10:1) to give a white solid (150.0 mg). 1 HNMR(400MHz,CDCl 3 )δ7.81(s,1H),7.62(s,1H),6.88(d,J＝12.0Hz,1H),6.04(d,J＝12.4Hz,1H),3.97(q,J＝7.2Hz,2H),1.80(s,6H),1.61(s,9H),1.38(t,J＝7.2Hz,3H)。
Synthesis of 2- (6-bromo-3, 3-dimethyl-1-oxoisoindol-4-yl) acetaldehyde (2):
(E) -5-bromo-7- (2-ethoxyvinyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (150.0 mg,0.37 mmol) was dissolved in tetrahydrofuran (3 mL), concentrated hydrochloric acid (1 mL) was added dropwise at room temperature and stirred for 3 hours. To the reaction system was added dropwise saturated aqueous sodium bicarbonate until ph=9. Extraction with ethyl acetate (5 mL. Times.3), washing the organic phase with saturated brine (10 mL), drying, and spin-drying gave a white crude product (80.0 mg). LC-MS: [ M+H ]] + :282.2,284.2。
Synthesis of 6-bromo-4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (3):
the compound 2- (6-bromo-3, 3-dimethyl-1-oxoisoindol-4-yl) acetaldehyde (80.0 mg,0.28 mmol) was dissolved in ethanol (4 mL), sodium borohydride (85.0 mg,2.25 mmol) was added at room temperature and reacted for 1 hour. The reaction was quenched with ice water (8 mL), extracted with ethyl acetate (8 mL. Times.3), the organic phase was washed with saturated aqueous sodium chloride (8 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a white crude product (80.0 mg). LC-MS: [ M+H ] ] + :284.1,286.1。
Synthesis of 4- (2-hydroxyethyl) -3, 3-dimethyl-6-boronic acid pinacol ester-isoindol-1-one (4):
the compound 6-bromo-4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (80.0 mg,0.28 mmol), pinacol biborate (85.0 mg,0.33 mmol), potassium acetate (70.0 mg,0.71 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (20.0 mg,0.03 mmol) was suspended in anhydrous dioxane (4 mL), reacted at 100℃for 18 hours under nitrogen protection, water (10 mL) was added to the system, followed by extraction with ethyl acetate (10 mL. Times.3), and the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a brown crude product (80.0 mg). LC-MS: [ M+H ]] + :332.0。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (common int-20):
the compound 4- (2-hydroxyethyl) -3, 3-dimethyl-6-boronic acid pinacol ester-isoindol-1-one (80.0 mg,0.24 mmol), 2, 4-dichloro-5-fluoropyrimidine (50.0 mg,0.30 mmol), potassium phosphate (125.0 mg,0.59 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (17.0 mg,0.02 mmol) was suspended in anhydrous dioxane (4 mL) and water (0.8 mL) under nitrogenThe reaction was carried out at 100℃for 2 hours. The reaction was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol=10:1) to give a brown solid (60.0 mg). LC-MS: [ M+H ] ] + :336.3。
Preparation example 18 Synthesis of common intermediate common int-21
Synthesis of 4-bromo-1-ethoxy-2-nitrobenzene (2):
4-bromo-1-fluoro-2-nitrobenzene (1.0 g,4.55 mmol) was dissolved in N, N-dimethylformamide (10 mL), and sodium ethoxide (619.3 mg,9.1 mmol) was added to the solution. The mixture was stirred at 50 ℃ for 12 hours until complete consumption of the starting material. The reaction mixture was cooled, quenched with water (10 mL), and extracted with ethyl acetate (50 mL. Times.3). The organic phases were combined, washed with saturated brine (15 mL. Times.3), finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=3:1) afforded a white solid (1.0 g). LC-MS: [ M+H ]] + :246.2。
Synthesis of 1- (4-ethoxy-3-nitrophenyl) -4-methylpiperazine (3):
to a solution of 4-bromo-1-ethoxy-2-nitrobenzene (1.0 g,4.06 mmol) in dioxane (10 mL) was added 1-methylpiperazine (529.2 mg,5.28 mmol), tris (dibenzylideneacetone) dipalladium (186.1 mg,0.20 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (166.9 mg,0.41 mmol) and cesium carbonate (2648.3 mg,8.13 mmol) in this order. The resulting mixture was stirred under nitrogen at 100 ℃ for 6 hours. After the reaction mixture was cooled, it was diluted with water (10 mL) and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=3:1) afforded a white solid (400.0 mg). LC-MS: [ M+H ] ] + :266.2。
Synthesis of 2-ethoxy-5- (4-methylpiperazin-1-yl) aniline (4):
1- (4-ethoxy-3-nitrophenyl) -4-methylpiperazine (200.0 mg,0.75 mmol) and palladium on carbon (80.2 mg,10% w/w) were dissolved in methanol (3 mL). Inserting hydrogen balloon and replacing threeSecondary hydrogen. The mixture was stirred at room temperature for 5 hours until the starting material was completely consumed. The reaction solution was concentrated by filtration to give a black solid (120.0 mg). LC-MS: [ M+H ]] + :236.2。
Preparation example 19 Synthesis of common intermediate common int-22
Synthesis of 6-bromo-4- (dimethylphosphoryl) -3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (2):
the compound 6-bromo-4-iodo-3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (1.3 g,2.88 mmol), dimethylphosphine oxide (292.0 mg,3.74 mmol), tris (dibenzylideneacetone) dipalladium (265.0 mg,0.29 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (335.0 mg,0.58 mmol), triethylamine (874.3 mg,8.64 mmol) were dissolved in 1, 4-dioxane (10 ml) under nitrogen and stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (80 mL. Times.2). The combined organic phases were washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (0-15% methanol/dichloromethane) to give a white solid (550.0 mg). LC-MS: [ M+H ] ] + :402.1,404.1。
Synthesis of 4- (dimethylphosphoryl) -3-methyl-1-oxo-6-boronic acid pinacol ester isoindoline-2-carboxylic acid tert-butyl ester (3):
under nitrogen, 6-bromo-4- (dimethylphosphoryl) -3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (530.0 mg,1.32 mmol), pinacol biborate (503.0 mg,1.98 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were reacted]Palladium dichloride (94.0 mg,0.13 mmol) and potassium acetate (388.0 mg,3.95 mmol) were placed in 1, 4-dioxane (6 ml) and reacted at 90℃for 10 hours. The reaction solution was concentrated by cooling to give a crude black solid (1.2 g) which was used directly in the next reaction. LC-MS: [ M+H ]] + :450.2。
Synthesis of tert-butyl 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (dimethylphosphoryl) -3-methyl-1-oxoisoindoline-2-carboxylate (common int-22):
under nitrogen, 4- (dimethylphosphoryl) -3-methyl-1-oxo-6-boronic acid pinacol ester isoindoline-2-carboxylic acid tert-butyl ester (1.2 g, crude product), 2, 4-dichloro-5-fluoropyrimidine (239.0 mg,1.43 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were added]Palladium dichloride (87.0 mg,0.12 mmol) and potassium phosphate (763.0 mg,3.59 mmol) were dissolved in 1, 4-dioxane (10 ml), and reacted at 90℃for 2 hours. The reaction mixture was diluted with water (50 mL), and extracted with ethyl acetate (100 mL. Times.2). The combined organic phases were washed with saturated sodium chloride solution (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (0-15% methanol/dichloromethane) to give a white solid (420.0 mg). LC-MS: [ M+H ] ] + :454.1. Preparation example 20 Synthesis of common intermediate common int-23
Synthesis of methyl 2-cyanothiophene-3-carboxylate (2):
methyl 2-bromothiophene-3-carboxylate (25.0 g,113.09 mmol) was dissolved in N-methylpyrrolidone (150 mL), cuprous cyanide (20.2 g,225.55 mmol) was added and the reaction stirred under nitrogen at 120℃for 18 hours. LCMS monitored reaction was complete. The reaction mixture was cooled to room temperature, diluted with water (1000 mL), and extracted with ethyl acetate (500 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=10:1) to give the product as a pale yellow solid (12.0 g). LC-MS: [ M+H ]] + :168.1。
Synthesis of spiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -4' (5 ' H) -one (3):
methyl 2-cyanothiophene-3-carboxylate (10.0 g,59.82 mmol) was dissolved in diethyl ether (600 mL), tetraisopropyl titanate (18.7 g,65.79 mmol) was added at room temperature, and then a solution of ethyl magnesium bromide in tetrahydrofuran (65.8 mL,2mol/L,131.60 mmol) was added dropwise. After the completion of the dropping, the reaction solution was stirred at room temperature for two hours. LCMS detected complete reaction. To the reaction mixture was added dilute aqueous hydrochloric acid (100 mL,1 mol/L), and the mixture was extracted with ethyl acetate (400 mL. Times.3). The combined organic phases were washed with saturated brine (300 mL) and the organic phase was separated using anhydrous sulfur Drying sodium acid, filtering and concentrating. The crude product was purified by column chromatography on silica gel (dichloromethane: ethyl acetate=1:1) to give a brown solid (2.2 g). LCMS [ M+H ]] + :166.2。
Synthesis of 2 '-bromospiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -4 '(5' H) -one (4):
spiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-4 '(5' h) -one (2.2 g,13.32 mmol) was dissolved in acetic acid/water=1.2:1 (20 mL) and cooled in an ice bath before slowly dropwise adding bromine (2.3 g,14.39 mmol). After the completion of the dropwise addition, the reaction solution was stirred for 0.5 hour. LCMS showed complete reaction. To the reaction solution was poured a saturated aqueous solution of sodium thiosulfate (30 mL), and the reaction was quenched, followed by extraction with ethyl acetate (100 mL. Times.3). The organic phases were combined and washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (dichloromethane: ethyl acetate=1:1) to give a tan solid (1.5 g). LCMS [ M+H ]] + :244.2,246.2。
Synthesis of 2' -bromo-4 ' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -5' (4'H) -carboxylic acid tert-butyl ester (5):
2 '-bromospiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-4 '(5' H) -one (1.5 g,6.14 mmol) was dissolved in dichloromethane (20 mL) and after cooling on ice di-tert-butyl dicarbonate (2.7 g,12.37 mmol) and triethylamine (1.2 g,11.86 mmol) were added, followed by the final addition of 4-dimethylaminopyridine (80.0 mg,0.65 mmol). The reaction solution was stirred at room temperature for 1 hour. LCMS showed complete reaction. The reaction solution was directly concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate=5:1) to obtain a pale yellow solid (930.0 mg). LCMS: [ M+H-tBu ] + :288.1,290.1。
Synthesis of 4' -oxo-2 ' -boronic acid pinacol ester spiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -5' (4'H) -carboxylic acid tert-butyl ester (6):
2' -bromo-4 ' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-tert-butyl 5' (4'H) -carboxylate (800.0 mg,2.32 mmol), potassium acetate (684.0 mg,6.97 mmol), pinacol biborate (708.0 mg,2.79 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (170.0 mg,0.23 mmol) was dissolved in anhydrous 1, 4-dioxane (8 mL). The reaction solution is stirred at 80 ℃ under the protection of nitrogenAfter stirring for 0.5 hours, LCMS showed complete reaction. The reaction solution was directly used for the next reaction. LCMS: [ M+H ] t Bu] + :336.2。
Synthesis of tert-butyl 2' - (2-chloro-5-fluoropyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -5' (4'H) -carboxylate (common int-23):
under the protection of nitrogen, 4' -oxo-2 ' -boric acid pinacol ester spiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-tert-butyl 5' (4'H) -carboxylate (300.0 mg,0.77 mmol), potassium phosphate (489.0 mg,2.30 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (56.1 mg,0.077 mmol) and 2, 4-dichloro-5-fluoropyrimidine (128.0 mg,0.77 mmol) were placed in 1, 4-dioxane and water (8 mL, 5:1), and the reaction was heated to 80℃and stirred for 2 hours. LCMS monitored reaction was complete. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (ethyl acetate: petroleum ether=2:1) to give a white solid (150.0 mg). LCMS: [ M+H-tBu ] + :340.3。
EXAMPLE 1 Synthesis of Compound PR-01-049
Synthesis of 1-isoindolone-6-boronic acid pinacol ester (2):
the compound 6-bromo-2, 3-dihydro-1H-isoindol-1-one (500 mg,2.36 mmol), pinacol biboronate (898 mg,3.54 mmol), potassium acetate (693 mg,7.07 mmol), 1-bis (diphenylphosphine) ferrocene palladium dichloride (PdCl) 2 dppf,176mg,0.24 mmol) in anhydrous dioxane (15 mL), N 2 The reaction was carried out at 100℃for 3 hours under protection, water (10 mL) was added to the system, followed by extraction with ethyl acetate (20 mL. Times.3), the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product, and purified by column chromatography (methanol: dichloromethane=1:100 to 1:50) to give a white solid (280 mg). LC-MS: [ M+H ]] + :260.0。
Synthesis of 6- (2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindol-1-one (PR-01-049):
the compound 1-isoindolone-6-boronic acid pinacol ester (180 mg,0.70 mmol), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2,323mg,0.83 mmol), 1' -bis (diphenylphosphine) ferrocene palladium dichloride (Pd (dppf) Cl) 2 51mg,0.07 mmol), potassium phosphate (427 mg,2.01 mmol) were dissolved in a mixed solution of 1, 4-dioxane (8 mL) and water (0.8 mL), the system was subjected to vacuum nitrogen substitution three times, heated to 100℃for reaction for 5 hours, and after the reaction was completed, the reaction was cooled to room temperature. The reaction solution was diluted with water (10 mL), the aqueous phase was extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined and washed successively with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was isolated as a white solid PR-01-049 (39.7 mg). 1 H NMR(400MHz,CD 3 OD)δ8.61(s,1H),8.54(d,J＝5.3Hz,1H),8.36(d,J＝8.0Hz,1H),8.30(d,J＝2.8Hz,1H),7.74(d,J＝8.0Hz,1H),7.47(d,J＝5.2Hz,1H),7.25(d,J＝8.4Hz,1H),6.76(dd,J＝9.0,2.8Hz,1H),4.55(s,2H),3.48-3.40(m,4H),3.19-3.10(m,4H),2.74(s,3H)；LC-MS:[M+H] + :485.2。
EXAMPLE 2 Synthesis of Compounds PR-01-075
3, 3-dimethyl-6- (2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindol-1-one (PR-01-075) synthesis:
synthesis of PR-01-075 (2.3 mg) as a white solid starting from 3, 3-dimethyl-6-boronic acid pinacol ester isoindol-1-one in a similar procedure as Compound PR-01-049. 1 H NMR(400MHz,CD 3 OD)δ8.53(d,J＝5.1Hz,1H),8.46(s,1H),8.40(d,J＝8.2Hz,1H),8.18(s,1H),7.72(d,J＝7.9Hz,1H),7.44(d,J＝5.3Hz,1H),7.21(d,J＝8.9Hz,1H),6.72(d,J＝8.9Hz,1H),3.27(m,4H),2.63(m,4H),2.34(s,3H),1.58(s,6H)；LCMS:[M+H] + :513.4。
EXAMPLE 3 Synthesis of Compound PR-01-066
Synthesis of (5-oxo-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-3-yl) boronic acid (2):
3-bromo-6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridin-5-one (50.0 mg,0.24 mmol), pinacol diboronate (78.7 mg,0.31 mmol) and potassium acetate (97.4 mg,0.99 mmol) were dissolved in 1, 4-dioxane (2 mL) and 1,1' -bis (diphenylphosphino) ferrocene palladium (II) dichloride (PdCl) 2 dppf,19.2mg,0.026 mmol) was added to the reaction solution, and replaced with nitrogen three times. The reaction solution was stirred at 80℃for 3 hours. LCMC detected complete reaction. The reaction was concentrated to give the crude product as a black solid (150 mg), which was used directly in the next step without further purification. LCMS [ M+H ]] + :179.1。
Synthesis of 3- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (PR-01-066):
5-oxo-6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridine-3-yl) boronic acid (150.0 mg,0.84 mmol), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2,200mg,0.52 mmol) was dissolved in 1, 4-dioxane (3 mL) and water (0.5 mL), and potassium carbonate (42.8 mg,0.31 mmol) and 1,1' -bis (diphenylphosphino) ferrocene palladium (II) dichloride (8.4 mg,0.01 mmol) were added to the reaction solution. The reaction system was replaced with nitrogen three times. The reaction solution was stirred at 80℃for 5 hours. LCMS detected complete reaction. The reaction mixture was poured into water (5 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, dried and concentrated, purified on a silica gel column (dichloromethane: methanol=100:1-6:1, rf=0.2) and then reversed phase (basic) to give white solid PR-01-066 (22.9 mg). 1 H NMR(400MHz,DMSO)δ9.47(d,J＝2.0Hz,1H),9.06(s,1H),8.92(s,1H),8.70(d,J＝2.0Hz,1H),8.58(d,J＝4.0Hz,1H),7.64(d,J＝4.0Hz,1H),7.53(d,J＝4.0Hz,1H),7.23(dd,J＝9.1,1.3Hz,1H),6.78(dd,J＝12.0,4.0Hz,1H),4.52(s,2H),3.20-3.11(m,4H),2.48-2.42(m,4H),2.21(s,3H)；LCMS:[M+H] + :486.2。
EXAMPLE 4 Synthesis of Compound PR-01-055
Synthesis of 5-bromo-2-methylthiophene-3-carboxylic acid methyl ester (2):
the compound methyl 2-methylthiophene-3-carboxylate (2.00 g,12.80 mmol) and NBS (2.70 g,15.17 mmol) were dissolved in DMF (20 mL) and reacted at room temperature for 16 hours. To the reaction solution was added water (15 mL), extracted with n-hexane (20 mL. Times.3), and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated by filtration to give a yellow oily compound (2.3 g). 1 H NMR(400MHz,CDCl 3 )δ7.34(s,1H),3.86(s,3H),2.69(s,3H)。
Synthesis of 5-bromo-2- (dibromomethyl) thiophene-3-carboxylic acid methyl ester (3):
the compound methyl 5-bromo-2-methylthiophene-3-carboxylate (1.00 g,4.25 mmol), NBS (1.90 g,10.68 mmol), azobisisobutyronitrile (AIBN, 70.6mg,0.43 mmol) was dissolved in carbon tetrachloride (10 mL) and heated to 80℃and stirred for 16 hours. To the reaction solution was added water (10 mL), extracted with dichloromethane (10 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was subjected to column chromatography (petroleum ether: ethyl acetate=100:1) to give a white solid (1.2 g). 1 H NMR(400MHz,CDCl 3 )δ7.82(s,1H),7.32(s,1H),3.89(s,3H)。
Synthesis of 5-bromo-2-formylthiophene-3-carboxylic acid methyl ester (4):
the compound methyl 5-bromo-2- (dibromomethyl) thiophene-3-carboxylate (1.20 g,3.05 mmol), silver nitrate (518.1 mg,3.05 mmol) were dissolved in ethanol (48.8 mL) and water (7.2 mL) heated to 65 ℃ for reaction for 6 hours. TLC monitors the disappearance of the raw materials, cools to room temperature, filters out solids and then collects filtrate for spin-drying to obtain crude products. Water (10 mL) was added, extracted with ethyl acetate (10 mL. Times.3), and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to give a yellow solid (700 mg). 1 H NMR(400MHz,CDCl 3 )δ10.41(s,1H),7.46(d,J＝9.7Hz,1H),3.88(s,3H)。
Synthesis of methyl 5-bromo-2- (3, 4-dimethoxybenzyl) amino) methylthiophene-3-carboxylate (5):
the compound methyl 5-bromo-2-formylthiophene-3-carboxylate (8238 mg,3.32 mmol), (3, 4-Dimethoxyphenyl) Methylamine (DMBNH) 2 837.7mg,5.01 mmol) in dichloromethane (16.6 mL) and glacial acetic acid (2.07 mL), warmed to 35℃and stirred for 30 min, after which sodium triacetoxyborohydride (2.1 g,9.91 mmol) was added and stirring continued for 18 h. To the reaction solution was added dichloromethane (5 mL), which was diluted with water (5 mL) and brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Purification by column chromatography (ethyl acetate: petroleum ether=10:1 to 5:2) afforded yellow solid 5 (624 mg). LC-MS: [ M+H ]] + :400.0。
Synthesis of 5-bromo-2- (3, 4-dimethoxybenzyl) amino) methylthiophene-3-carboxylic acid (6):
the compound methyl 5-bromo-2- (3, 4-dimethoxybenzyl) amino) methylthiophene-3-carboxylate (624 mg,1.56 mmol) was dissolved in methanol (1 mL), 5M NaOH (3.1 mL,15.59 mmol) was added, and the mixture was stirred at room temperature for 5 hours. Methanol in the reaction mixture was spun off, diluted with water (2 mL), ph=5-6 was adjusted with 6N hydrochloric acid, and a solid was precipitated, filtered, and dried under vacuum to give a yellow solid (510 mg). LC-MS: [ M+H ]] + :386.0。
Synthesis of 2-bromo-5- (3, 4-dimethoxybenzyl) -5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (7):
the compound 5-bromo-2- (3, 4-dimethoxybenzyl) amino) methylthiophene-3-carboxylic acid (510 mg,1.32 mmol), 1-hydroxybenzotriazole (268 mg,1.98 mmol) and TEA (400 mg,3.95 mmol) were dissolved in DMF (10 mL), heated to 50℃and stirred for 10 min, then EDCI (380 mg,1.98 mmol) was added and stirred for 3 h at 50 ℃. The reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 ml×2) and saturated brine (5 ml×2), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=6:1 to 3:1) to give a yellow oil (250 mg). 1 H NMR(400MHz,CDCl 3 )δ7.24(s,1H),7.20(d,J＝8.0Hz,1H),6.49-6.41(m,2H),4.67(s,2H),4.28(s,2H),3.83(s,3H),3.79(s,3H)。
Synthesis of 5- (3, 4-dimethoxybenzyl) -2-boronic acid pinacol ester-5, 6-dihydro-4H-thiophene [2,3-c ] pyrrol-4-one (8):
the compound 2-bromo-5- (3, 4-dimethoxybenzyl) -5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (150 mg,0.41 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (32 mg,0.044 mmol), pinacol diboronate (155 mg,0.61 mmol) and potassium acetate (44 mg,0.44 mmol) were dissolved in 1, 4-dioxane (2 mL) and heated to 80℃under nitrogen for 3 hours. The reaction solution of the resulting compound was directly used in the next reaction. LC-MS: [ M+H ]] + :416.0。
Synthesis of 5- (3, 4-dimethoxybenzyl) -2- (2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-thiophene [2,3-c ] pyrrol-4-one (9)
4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2,85.3mg,0.22 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were added to the reaction solution]Palladium (II) dichloride (26 mg,0.036 mmol), potassium carbonate (99.5 mg,0.72 mmol) and water (2 ml) were heated to 80℃under nitrogen to react for 3 hours. Ethyl acetate (5 mL) was added to dilute, water (5 ml×2) was used to wash, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by column chromatography (methanol: dichloromethane=0:100 to 1:10) to give a black oil (73 mg). LC-MS: [ M+H ] ] + :641.0。
Synthesis of 2- (2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-thiophene [2,3-c ] pyrrol-4-one (PR-01-055):
the compound 5- [ (3, 4-dimethoxyphenyl) methyl]-2- (2- { [5- (4-methylpiperazin-1-yl) -2- [ (trifluoromethyl) oxy)]Phenyl group]Amino } pyrimidin-4-yl) -5, 6-dihydro-4H-thiophene [3,2-c]Pyrrole-4-one (60 mg,0.094 mmol) and triethylsilane (144.2 mg,1.24 mmol) were dissolved in trifluoroacetic acid (0.8 mL), and the mixture was stirred at 50℃for 2 hours. The reaction solution was concentrated to dryness by an oil pump, and a white solid PR-01-055 (0.7 mg) was obtained by preparative separation. 1 H NMR(400MHz,CDCl 3 )δ8.48(d,J＝5.1Hz,1H),8.30(d,J＝2.8Hz,1H),7.85(s,1H),7.43(s,1H),7.16(d,J＝8.4Hz,1H),7.11(d,J＝5.2Hz,1H),6.57(dd,J＝9.0,2.7Hz,1H),6.05(s,1H),4.59(s,2H),3.39-3.29(m,4H),2.67-2.56(m,4H),2.38(s,3H)；LC-MS(ESI):(M+H) + 491.4。
EXAMPLE 5 Synthesis of Compound PR-01-054
Synthesis of 2- (2-hydroxypropan-2-yl) thiophene-3-carboxylic acid (2):
thiophene-3-carboxylic acid (500.0 mg,3.90 mmol) was dissolved in tetrahydrofuran (20 mL), the nitrogen was purged three times, the system was cooled to-70℃and n-butyllithium (6 mL,1.6M cyclohexane solution, 9.60 mmol) was slowly added dropwise to the system while keeping the temperature below-55℃and the reaction was continued with stirring at-70℃for 1 hour after the completion of the dropwise addition, then acetone (0.38 mL,5.17 mmol) was slowly added and the system temperature was raised to 0℃and the reaction was continued with stirring at this temperature for 3 hours, and after the completion of the reaction, 4M hydrochloric acid (3 mL) was added dropwise and the reaction was continued with stirring at room temperature overnight. After the reaction was complete, the reaction mixture was filtered through a celite pad and the solid was washed with toluene. The filtrate was concentrated under reduced pressure to give a yellow liquid (434.0 mg).
Synthesis of 5-acetyl-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (3):
2- (2-hydroxy-propan-2-yl) thiophene-3-carboxylic acid (200.0 mg,1.07 mmol) was dissolved in acetonitrile (8 mL), the mixture was evacuated and nitrogen was exchanged three times, boron trifluoride diethyl ether (0.16 mL,1.29 mmol) was added to the system at room temperature, and the mixture was heated to 60℃to react for 16 hours. After the reaction was completed, the solvent was removed by spin-chromatography (petroleum ether: ethyl acetate=10:1, rf=0.6) to give a white solid (75.0 mg). LC-MS: [ M+H ]] + :210.0。
Synthesis of 6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (4):
the compound 5-acetyl-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (500.0 mg,2.39 mmol) and sodium hydroxide (286.8 mg,7.17 mmol) were dissolved in a mixed solution of methanol (15 mL) and water (15 mL), and stirred at room temperature for 2 hours. After the reaction was completed, the pH was adjusted to=5 by adding 1N hydrochloric acid solution, extracted three times with ethyl acetate (15 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentratedCrude products are obtained. Column chromatography (dichloromethane: methanol=10:1, rf=0.4) afforded a white solid (320.0 mg). LC-MS: [ M+H ]] + :168.1。
Synthesis of 2-bromo-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (5):
The compound 6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (170.0 mg,1.02 mmol) was dissolved in a mixed solution of acetic acid (2.8 mL) and water (2.2 mL), the system was cooled to 0℃and bromine (179.0 mg,1.12 mmol) was slowly added dropwise to the system, and after the dropwise addition was completed, the reaction was continued at 0℃for 0.5 hours. After the reaction was completed, the reaction was quenched by adding water (3 mL), extracted three times with ethyl acetate, the organic phases were combined, washed with saturated aqueous sodium sulfite solution, then with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (dichloromethane: methanol=10:1, rf=0.3) gave a tan solid (230.0 mg). LC-MS: [ M+H ]] + :245.9。
Synthesis of 6, 6-dimethyl-2-boronic acid pinacol ester-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (6):
the compound 2-bromo-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (100.0 mg,0.41 mmol), potassium acetate (119.5 mg,1.22 mmol), 1' -bis (diphenylphosphino) ferrocene palladium (II) dichloride (29.7 mg,0.04 mmol), pinacol ester (134.0 mg,0.53 mmol) were dissolved in 1, 4-dioxane (3 mL), the system was evacuated to three nitrogen changes, heated to 90℃for 2 hours, and the reaction was allowed to cool to room temperature. The reaction mixture was diluted with water (10 mL), the aqueous phase was washed with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed successively with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. Column chromatography (dichloromethane: methanol=10:1, rf=0.3) gave a white solid (26.0 mg). LC-MS: [ M+H ] ] + :294.1。
Synthesis of 6, 6-dimethyl-2- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-thiophene [2,3-c ] pyrrol-4-one (PR-01-054):
the compound 6, 6-dimethyl-2-boronic acid pinacol ester-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (26.0 mg,0.09 mmol), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2,41.5mg,0.11 mmol), 1' -bis (diphenylphosphino) ferrocene palladium (II) dichloride (6.5 mg,0.01 mmol), potassium phosphate (57.3 mg,0.27 mmol) were dissolved in a mixed solution of 1, 4-dioxane (1 mL) and water (0.1 mL), the system was evacuated to change nitrogen three times, heated to 90℃for 2 hours, and after the reaction was completed, the reaction was cooled to room temperature. The reaction mixture was diluted with water (10 mL), the aqueous phase was extracted with ethyl acetate (10 mL. Times.3), the combined organic phases were successively washed with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude product, which was isolated as a white solid PR-01-054 (6.8 mg). 1 H NMR(400MHz,DMSO)δ8.84(s,1H),8.62(s,1H),8.48(d,J＝5.2Hz,1H),8.05(s,1H),7.66(d,J＝2.8Hz,1H),7.47(d,J＝5.2Hz,1H),7.21(dd,J＝9.0,1.3Hz,1H),6.73(dd,J＝9.1,3.0Hz,1H),3.21(s,4H),2.47(d,J＝4.8Hz,4H),2.22(s,3H),1.52(s,6H)；LC-MS:[M+H] + :519.2。
EXAMPLE 6 Synthesis of Compounds PR-01-070
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) isoindol-1-one (2):
the compound 6-pinacol borate isoindol-1-one (500 mg,1.93 mmol), 2, 4-dichloro-5-fluoropyrimidine (451.1 mg, 2.704 mmol), bis triphenylphosphine palladium dichloride (135.4 mg,0.193 mmol) and potassium phosphate (1.20 g,5.65 mmol) were dissolved in 1, 4-dioxane (3 mL), heated to 80 ℃ under nitrogen protection, reacted for 16 hours, cooled, dichloromethane (20 mL) was added to the reaction mixture to dilute and filter, the organic phase was washed with water, the combined organic phases were dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (dichloromethane/methanol: 10/1 rf=0.6). After dissolving the product in 5mL of dichloromethane, 20mL of ethyl acetate were added and the solvent was evaporated in a water bath at 40 ℃ until a solid precipitated, which was cooled and filtered to give the product as a pink solid (180 mg). LC-MS: [ M+H ] ] + :264.0。
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindol-1-one (PR-01-070):
the compound 6- (2-chloro-5-fluoropyrimidin-4-yl) isoindol-1-one (160 mg,0.61 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1,250.6mg,0.91 mmol), palladium acetate (13.62 mg,0.061 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (75.6 mg,0.12 mmol) and cesium carbonate (593.2 mg,1.82 mmol) were dissolved in 1, 4-dioxane (2 mL), heated to 100℃under nitrogen protection and reacted for 3 hours, the reaction solution was cooled, ethyl acetate (10 mL) was added to the reaction solution to dilute, water and saturated brine were used, the organic phase was collected and dried over anhydrous sodium sulfate, and the reaction solution was separated to give light yellow solid PR-01-070 (36.1 mg). 1 H NMR(400MHz,DMSO)δ9.07(s,1H),8.73(s,1H),8.64(d,J＝3.5Hz,1H),8.28(s,1H),8.24(d,J＝8.1Hz,1H),7.76(d,J＝8.1Hz,1H),7.55(d,J＝2.9Hz,1H),7.21(d,J＝7.9Hz,1H),6.75(dd,J＝9.1,2.9Hz,1H),4.48(s,2H),3.18-3.14(m,4H),2.48-2.42(m,4H),2.21(s,3H)；LC-MS:[M+H] + :503.1。
EXAMPLE 7 Synthesis of Compound PR-01-069
Synthesis of 6- (5-chloro-2- (methylthio) pyrimidin-4-yl) -isoindol-1-one (2):
the compound 4, 5-dichloro-2- (methylthio) pyrimidine (600 mg,3.08 mmol), pinacol 6-borate-isoindol-1-one (956.4 mg,3.70 mmol), sodium carbonate (815.1 mg,7.70 mmol), 1' -bis-diphenylphosphino ferrocene palladium dichloride (179.9 mg,0.25 mmol) were dissolved in 1, 4-dioxane (6 mL) and water (1.5 mL), reacted for 2 hours at 80℃under nitrogen protection, and the reaction mixture was concentrated and purified by column chromatography (methanol: dichloromethane=1:200-1:100, rf=0.4) to give a white solid (650 mg). LC-MS: [ M+H ] ] + :292.0。
Synthesis of tert-butyl 6- (5-chloro-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (3):
the compound 6- (5-chloro-2- (methylthio) pyrimidine-4-yl) isoindol-1-one (650 mg,2.23 mmol), 4-dimethylaminopyridine (544.4 mg,4.46 mmol), triethylamine (451.3 mg,4.46 mmol) in dichloromethane (8 mL) and di-tert-butyl dicarbonate (972.5 mg,4.46 mmol) were added at 0deg.C and reacted for 2 hours at 25deg.C. 10mL of water was added, dichloromethane (10 mL. Times.3) was added, the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by column chromatography (petroleum ether: ethyl acetate=10:1-5:1, rf=0.3) to give white solid 3 (670 mg). LC-MS [ M-t-Bu+H ]] + :336.0。
Synthesis of tert-butyl 6- (5-chloro-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (4):
the compound 6- (5-chloro-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylic acid tert-butyl ester (670 mg,1.71 mmol) was dissolved in dichloromethane (15 mL), m-chloroperoxybenzoic acid (1.18 g,6.84 mmol) was slowly added at 0 ℃ and reacted at room temperature for 2 hours, saturated sodium bicarbonate solution (3 mL) was added to the reaction solution, 5mL of water was added, dichloromethane (5 ml×3) was added, the extracts were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by column chromatography (petroleum ether: ethyl acetate=5:1-3:1, rf=0.2) to give a white solid (670 mg). LC-MS [ M-Boc+H ] ] + :324.0。
Synthesis of tert-butyl 6- (5-chloro-2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxyphenyl) amino) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylate (5):
to N- [5- (4-methylpiperazin-1-yl) -2- [ (trifluoromethyl) oxy ]]Phenyl group]Cesium carbonate (115 mg,0.35 mmol) was added to a suspension of formamide (common int-3,107mg,0.35 mmol) and tert-butyl 6- (5-chloro-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (100 mg,0.24 mmol) in dry DMF (4 mL). The mixture was stirred at 70℃for 2 hours. The reaction mixture was treated with MeCN/H 2 C18 reverse phase column purification of o=1:1 gave the product (70 mg) as a yellow solid. LC-MS: [ M+H ]] + :619.2.
Synthesis of 6- (5-chloro-2- (5-methylpiperazin-1-yl) -2- (trifluoromethoxyphenyl) amino) pyrimidin-4-yl) -isoindol-1-one (PR-01-069):
to the 6- (5-chloro-2- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy)Phenyl) amino) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylic acid tert-butyl ester (62 mg,0.10 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (0.5 mL,6.73 mmol) was slowly added and stirred at room temperature for 6 hours. The mixture was purified by high performance liquid chromatography to give the product PR-01-069 (37 mg) as a white solid. 1 H NMR(400MHz,DMSO)δ9.30(s,1H),8.72(s,1H),8.62(s,1H),8.06(s,1H),8.01(d,J＝4.0,1H),7.73(d,J＝8.0Hz,1H),7.49(d,J＝4.0Hz,1H),7.19(d,J＝12.0Hz,1H),6.75(dd,J＝8.0,4.0Hz,1H),4.48(s,2H),3.18-3.06(m,4H),2.46-2.36(m,4H),2.21(s,3H).LC-MS[M+H] + :519.1。
EXAMPLE 8 Synthesis of Compound PR-01-068
Synthesis of 6- (5-methyl-2- (methylthio) pyrimidin-4-yl) isoindol-1-one (2):
4-chloro-5-methyl-2- (methylthio) pyrimidine (200 mg,1.15 mmol) and 6-boronic acid pinacol ester isoindol-1-one (297.8 mg,1.15 mmol) are dissolved in 1, 4-dioxane (5 mL) and water (0.5 mL), followed by potassium carbonate (317.9 mg,2.30 mmol) and (1, 1' -bis (diphenylphosphino) ferrocene) palladium dichloride (Pd (dppf) Cl 2 87.8mg,0.12 mmol) was added to the reaction solution, and the mixture was purged three times with nitrogen. The reaction solution was stirred at 80℃for 2 hours. LCMS checked for completion of the reaction, cooled to room temperature, and the reaction mixture was poured into water (5 mL) and extracted with dichloromethane (10 ml×3). The organic phases were combined, washed once with 10mL of brine, separated, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was subjected to column chromatography (ethyl acetate: petroleum ether=0-75%) to give a brown solid (190 mg). LC-MS: [ M+H ]] + :272.1。
Synthesis of 6- (5-methyl-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylic acid tert-butyl ester (3):
the compound 6- (5-methyl-2- (methylthio) pyrimidin-4-yl) isoindol-1-one (190.0 mg,0.70 mmol), triethylamine (212.5 mg,2.1 mmol), 4-dimethylaminopyridine (DMAP, 52.5mg,0.43 mmol) was dissolved in dichloromethane (5.0 mL) and di-tert-butyl dicarbonate ((Boc) was added at 0deg.C 2 O,93.4mg,0.43mmol), stirring for 5 hours at room temperature. The TLC plate detects that the starting material was reacted (petroleum ether: ethyl acetate=1:1, rf=0.4), water (5 mL) was added to the reaction solution, the organic phase was collected and dried over anhydrous sodium sulfate, and after concentration by filtration, a crude product was obtained by column chromatography (ethyl acetate: petroleum ether=0-30%) to give a yellow solid (200.0 mg). LC-MS: [ M+H ] ] + :372.1。
Synthesis of 6- (5-methyl-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylic acid tert-butyl ester (4):
the compound 6- (5-methyl-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylic acid tert-butyl ester (200.0 mg,0.54 mmol) was dissolved in dichloromethane (5 mL), m-chloroperoxybenzoic acid (m-CPBA, 279.6mg,1.62 mmol) was added at 0 ℃ and then stirred for 6 hours at room temperature, the reaction was detected by TLC plates (petroleum ether: ethyl acetate=3:1, rf=0.2), saturated sodium bicarbonate solution (5 mL) was added to the reaction solution, extracted with dichloromethane (5 ml×3), the organic phase was collected and dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was subjected to column chromatography (ethyl acetate: petroleum ether=0-40%) to give a white solid (189 mg). LC-MS [ M-H ]] - :402.0。
Synthesis of tert-butyl 6- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylate (5):
the compound 6- (5-methyl-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylic acid tert-butyl ester (139.0 mg,0.35 mmol), N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) carboxamide (common int-3,124.3mg,0.41 mmol), cesium carbonate (168.4 mg,0.52 mmol) was dissolved in ultra-dry N, N-dimethylformamide (2 mL) and heated to 80℃and stirred for 4 hours. LCMS detected completion of the reaction, water (5 mL) was added to the reaction solution after completion of the reaction, ethyl acetate was extracted (10 ml×3), the combined organic phases were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated by rotary evaporation to give crude product, which was subjected to reverse-phase column chromatography (acetonitrile: water=5-85%) to give a yellow solid (88 mg). LC-MS [ M+H ] ] + :599.0。
Synthesis of 6- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindol-1-one (PR-01-068):
the compound tert-butyl 6- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxoisoindole-2-carboxylate (83.0 mg,0.14 mmol) was dissolved in dichloromethane (2.5 mL) and trifluoroacetic acid (0.5 mL,6.7 mmol) was added and stirred at room temperature for 4 hours. LCMS detected completion of the reaction, the reaction was dried by oil pump, and a white solid (41 mg) was prepared. 1 H NMR(400MHz,CDCl3)δ8.44(d,J＝2.9Hz,1H),8.40(s,1H),8.16(s,1H),7.95(dd,J＝7.9,1.4Hz,1H),7.61(d,J＝7.9Hz,1H),7.41(s,1H),7.13(dd,J＝9.0,1.3Hz,1H),6.77(s,1H),6.50(dd,J＝9.0,2.9Hz,1H),4.55(s,2H),3.28-3.07(m,4H),2.60-2.49(m,4H),2.34(d,J＝2.4Hz,6H)；LC-MS:[M+H] + :499.7。
EXAMPLE 9 Synthesis of Compound PR-01-076
Synthesis of 5- (pinacol borate) -2H-isoindole-1, 3-dione (2):
5-bromo-2H-isoindole-1, 3-dione (400.0 mg,1.77 mmol), pinacol diboronate (675.5 mg,2.66 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]A mixture of palladium dichloride dichloromethane complex (145.9 mg,0.18 mmol) and potassium acetate (347.4 mg,3.54 mmol) in dioxane (8 mL) was stirred at 90℃for 4 hours. The reaction mixture was diluted with ethyl acetate (10 mL) and washed with water (12 mL). The organic phase was concentrated and purified by column on silica gel (petroleum ether/ethyl acetate=4:1, rf=0.2) to give the product as a yellow solid (300 mg). LC-MS: [ M+H ]] + :274.1. Synthesis of 2-carbamoyl-4- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) benzoic acid (3):
5-boronic acid pinacol ester) -2H-isoindole-1, 3-dione (100 mg,0.37 mmol), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (143.5 mg,0.37 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]A mixture of palladium dichloride dichloromethane complex (30.0 mg,0.037 mmol) and potassium carbonate (101 mg,0.73 mmol) was suspended in dioxane-water (3:1, 4 mL), and the reaction solution was stirred under nitrogen at 70℃for 4 hours and then concentrated directly.The crude product was purified by reverse phase column chromatography to give the product as a yellow solid (80 mg). LC-MS: [ M+H ]] + :517.1. Synthesis of 5- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindole-1, 3-dione (PR-01-076):
to a solution of 2-carbamoyl-4- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) benzoic acid (80 mg,0.15 mmol), N, N' -carbonyldiimidazole (37.3 mg,0.23 mmol) in tetrahydrofuran (2 mL) was slowly added triethylamine (47.6 mg,0.47 mmol) and stirred at room temperature for 16 hours. The reaction solution was concentrated, and then purified to give a pale yellow solid product (29 mg). LC-MS: [ M+H ]] + :499.1。
EXAMPLE 10 Synthesis of Compound PR-01-067
Synthesis of 6-bromo-3-methyl-pyridine-2-carboxylic acid ethyl ester (2):
The compound 6-bromo-3-methylpyridine carboxylic acid (1.0 g,4.63 mmol), potassium carbonate (1.6 g,11.57 mmol) and iodoethane (867.2 mg,5.56 mmol) were dissolved in N, N-dimethylformamide (10 mL). The reaction was stirred at 50℃overnight. LCMS and TLC detected complete reaction. The reaction was quenched with saturated ammonium chloride solution (40 mL), the system was extracted three times with ethyl acetate (30 ml×3), the organic phases were combined, washed successively with water and saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product, which was subjected to column chromatography (ethyl acetate: petroleum ether=1:4, rf=0.5) to give a pale yellow oily product (0.96 g). LC-MS: [ M+H ]] + :244.0,246.0。
Synthesis of ethyl 6-bromo-3- (bromomethyl) picolinate (3):
the compound 6-bromo-3-methylpyridine-2-carboxylic acid ethyl ester (800.0 mg,3.28 mmol) was dissolved in carbon tetrachloride (10 mL), N-bromosuccinimide (583.8 mg,3.28 mmol) was added, azobisisobutyronitrile (AIBN, 54.2mg,0.33 mmol) was stirred overnight in an oil bath at 80℃and the reaction was complete by LCMS and TLC. Then quenched with saturated ammonium chloride solution (40 mL), extracted three times with ethyl acetate (20 mL. Times.3), combined withThe organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was subjected to column chromatography (ethyl acetate: petroleum ether=1:4, rf=0.4) to give a pale yellow oily product (700.0 mg). LC-MS: [ M+H ] ] + :321.9,323.9。
Synthesis of 2-bromo-5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (4):
the compound ethyl 6-bromo-3- (bromomethyl) picolinate (700.0 mg,2.17 mmol) was dissolved in 7M methanolic ammonia (10 mL) and reacted at room temperature for 1 hour, with complete reaction as detected by LCMS and TLC. The precipitated solid was filtered and dried to give a white solid product (350.0 mg), LC-MS: [ M+H ]] + :213.0,215.0。
Synthesis of tert-butyl 2-bromo-7-oxo-5, 7-dihydro-6H-pyrrolo [3,4-b ] pyridine-6-carboxylate (5):
the compound 2-bromo-5, 6-dihydro-7H-pyrrolo [3,4-b]Pyridin-7-one (300 mg,1.41 mmol) was dissolved in N, N-dimethylformamide (6 mL), di-tert-butyl dicarbonate (460.0 mg,2.11 mmol), triethylamine (285.4 mg,2.82 mmol), 4-dimethylaminopyridine (17.2 mg,0.14 mmol) was added and stirred at 50℃for 1 hour and the reaction was complete as detected by LCMS and TLC. Then quenched with saturated ammonium chloride solution (40 mL), the system was extracted with ethyl acetate (30 ml×3), the organic phases were combined, washed with water and saturated sodium chloride solution in this order, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was subjected to column chromatography (methanol: dichloromethane=1:20, rf=0.6) to give a pale yellow solid product (430.0 mg). LC-MS: [ M+H-Boc ]] + :213.0,215.0。
Synthesis of 7-oxo-2- (trimethylstannyl) -5, 7-dihydro-6H-pyrrolo [3,4-b ] pyridine-6-carboxylic acid tert-butyl ester (6):
The compound 2-bromo-7-oxo-5, 7-dihydro-6H-pyrrolo [3,4-b]Pyridine-6-carboxylic acid tert-butyl ester (300.0 mg,0.96 mmol) was dissolved in dry toluene (5 mL), hexamethylditin (629 mg,1.92 mmol) and tetraphenylpalladium phosphate (115.6 mg,0.10 mmol) were added, nitrogen was replaced three times, stirred at 100deg.C for 1.5h, and the reaction was complete by LCMS and TLC. Concentrated, and the plate was climbed to give a yellow oily product (160.0 mg). LC-MS: [ M+H ]] + :399.0。
Synthesis of tert-butyl 2- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -7-oxo-5, 7-dihydro-6H-pyrrolo [3,4-b ] pyridine-6-carboxylate (7):
the compound 7-oxo-2- (trimethylstannyl) -5, 7-dihydro-6H-pyrrolo [3,4-b]Pyridine-6-carboxylic acid tert-butyl ester (160 mg,0.40 mmol) was dissolved in toluene (5 mL), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (155.1 mg,0.40 mmol), tetrakis triphenylphosphine palladium (46.2 mg,0.04 mmol), nitrogen substitution three times, stirred at 100deg.C for 16 hours, then quenched with saturated ammonium chloride solution (10 mL), the system extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed sequentially with water and saturated sodium chloride solution, the organic phases dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative TLC to give a white solid (50.0 mg). LC-MS: [ M+H ] ] + :586.2。
Synthesis of 2- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (PR-01-067):
the compound 2- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -7-oxo-5, 7-dihydro-6H-pyrrole [3,4-b]Pyridine-6-carboxylic acid tert-butyl ester (50 mg,0.085 mmol) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (1 mL) was added, stirred for 1 hour at 25℃and then quenched with saturated sodium bicarbonate solution (10 mL), the system was extracted three times with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed successively with water and saturated sodium chloride solution, the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give a white solid (7.3 mg). LC-MS: [ M+H ]] + :486.1。 1 H NMR(400MHz,DMSO)δ9.10(s,1H),9.03(s,1H),8.65(d,J＝5.2Hz,1H),8.46(d,J＝8.0Hz,1H),8.23(d,J＝8.0Hz,1H),7.78(d,J＝5.2Hz,1H),7.58(d,J＝2.8Hz,1H),7.23(dd,J＝9.2,1.6Hz,1H),6.78(dd,J＝8.8,2.8Hz,1H),4.49(s,2H),3.21–3.16(m,4H),2.48–2.43(m,4H),2.23(s,3H)；LC-MS:[M+H] + :486.1。
EXAMPLE 11 Synthesis of Compound PR-01-071
Synthesis of 2- (methylthio) -4- (3-oxoisoindol-5-yl) pyrimidine-5-carbonitrile (2):
4-chloro-2- (methylthio) pyrimidine-5-carbonitrile (1.00 g,5.39 mmol), 6- (pinacol borate) isoindol-1-one (1.5 g,5.79 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (437.7 mg,0.54 mmol) and potassium carbonate (2.2 g,15.91 mmol) were dissolved in 1, 4-dioxane (20 mL) and the reaction liquid nitrogen was replaced three times. The reaction was stirred at 100℃overnight. LCMS detected complete reaction. The reaction solution was concentrated and purified by column chromatography (methanol: dichloromethane=0-10%, methanol: dichloromethane=1:10, rf=0.5) to give a product as a brown solid (1.35 g). LCMS [ M+H ] ] + :283.2。
Synthesis of tert-butyl 6- (5-cyano-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (3):
2- (methylthio) -4- (3-oxoisoindolin-5-yl) pyrimidine-5-carbonitrile (1.35 g,4.78 mmol) was dissolved in dichloromethane (20 mL) and cooled down in an ice bath, followed by the addition of di-tert-butyl dicarbonate (1.57 g,7.19 mmol), triethylamine (0.97 g,9.59 mmol) and 4-dimethylaminopyridine (58.6 mg,0.48 mmol) in sequence. After the reaction was gradually warmed to room temperature and stirred for 3 hours, LCMS showed complete reaction. The reaction solution was poured into water (20 mL), extracted with dichloromethane (20 mL. Times.3), the organic phases were combined and washed with saturated sodium chloride (15 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was subjected to column chromatography (ethyl acetate: petroleum ether=0-50%, ethyl acetate: petroleum ether=1:1, rf=0.5) to give a pale yellow solid (1.1 g). LCMS [ M-t-Bu+H ]] + :327.3。
Synthesis of tert-butyl 6- (5-cyano-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxodihydro-isoindole-2-carboxylate (4):
6- (5-cyano-2- (methylthio) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylic acid tert-butyl ester (450 mg,1.18 mmol) was dissolved in THF (5 mL) and water (5 mL), and potassium hydrogen peroxymonosulfate complex (2.17 g,6.27 mmol) was added to the reaction solution. The reaction was stirred at room temperature overnight and LCMS showed complete reaction. The reaction solution was poured into water (20 mL) and then extracted with ethyl acetate (10 ml×3), the organic phases were combined and washed with saturated sodium chloride (15 mL), dried over anhydrous sodium sulfate, and concentrated to give a crude product as a pale yellow solid (300 mg). LCMS [ M-t-Bu+H ] ] + :359.3。
Synthesis of tert-butyl 6- (5-cyano-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxodihydro-isoindole-2-carboxylate (5):
n- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) carboxamide (300 mg,0.99 mmol), cesium carbonate (644.6 mg,1.98 mmol) was dissolved in N, N-dimethylformamide (5 mL), the reaction solution was left at-10℃and tert-butyl 6- (5-cyano-2- (methylsulfonyl) pyrimidin-4-yl) -1-oxoisoindoline-2-carboxylate (491.9 mg,1.19 mmol) was dissolved in N, N-dimethylformamide (5 mL), followed by addition to the reaction solution. The reaction was stirred at-10 ℃ for 3 hours, LCMS showed about 10% product formation but the starting material disappeared. The reaction mixture was poured into water (30 mL), extracted with ethyl acetate (15 mL. Times.3), and the organic phase was washed with saturated brine and separated. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography (ethyl acetate: petroleum ether=0-50%, ethyl acetate: petroleum ether=1:1, rf=0.6) to give a yellow solid (30 mg). LCMS [ M+H ]] + :610.3。
Synthesis of 2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -4- (3-oxoisoindolin-5-yl) pyrimidine-5-carbonitrile (PR-01-071):
6- (5-cyano-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxodihydro-isoindole-2-carboxylic acid tert-butyl ester (30 mg,0.05 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (0.2 mL) was added dropwise to the reaction solution. The reaction solution was stirred at room temperature for one hour. LCMS showed complete reaction. The reaction solution was concentrated, followed by purification to give a white solid (2.2 mg). 1 H NMR(400MHz,DMSO)δ10.15(s,1H),8.93(s,1H),8.75(s,1H),8.19(s,1H),8.12(d,J＝7.6Hz,1H),7.78(d,J＝8.0Hz,1H),7.36-7.17(m,2H),6.88-6.85(m,1H),4.49(s,2H),3.17-3.15(m,4H),2.45-2.33(m,4H),2.21(s,3H)；LCMS:[M+H] + :510.4。
EXAMPLE 12 Synthesis of Compound PR-01-095
Synthesis of 3, 3-dimethyl-6- (pinacol borate) isoindol-1-one (2):
to a suspension of 6-bromo-3, 3-dimethylisoindol-1-one (250 mg,1.04 mmol), pinacol diboronate (396.1 mg,1.56 mmol) in dioxane (8 mL) was added [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (81.1 mg,0.10 mmol) and potassium acetate (204.1 mg,2.08 mmol). The reaction system was stirred at 80 ℃ for 4 hours under nitrogen protection and then concentrated directly to dryness. The crude product was purified by column on silica gel (ethyl acetate: petroleum ether=1:8, rf=0.2) to give a brown solid (280 mg). LC-MS: [ M+H ]] + :288.1。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (3):
to a stirred suspension of 3, 3-dimethyl-6- (pinacol borate) isoindol-1-one (100 mg,0.35 mmol) and 2, 4-dichloro-5-fluoropyrimidine (58.4 mg,0.35 mmol) in dioxane and water (3:1, 8 mL) under nitrogen was added [1,1' -bis (diphenylphosphine) ferrocene ]Palladium dichloride dichloromethane complex (28.4 mg,0.035 mmol) and potassium carbonate (96.6 mg,0.70 mmol). Stirring at 70℃for 2 hours under nitrogen protection and concentrating directly to dryness. The crude product was purified by column on silica gel (ethyl acetate: petroleum ether=1:5, rf=0.3) to give the product as a yellow solid (100 mg). LC-MS: [ M+H ]] + :292.0,294.0。
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-095):
to a suspension of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (100 mg,0.34 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (104 mg,0.38 mmol) and cesium carbonate (223 mg,0.68 mmol) in dioxane (6 mL) was added 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (BINAP, 43.6mg,0.07 mmol) and palladium acetate (7.6 mg,0.034 mmol) under nitrogen. The reaction was stirred at 100 ℃ for 16 hours under nitrogen protection and then concentrated directly to dryness. The crude product was purified by preparative liquid chromatography to give a white solid (31 mg). 1 H NMR(400MHz,DMSO)δ9.07(s,1H),8.82(s,1H),8.64(d,J＝3.2Hz 1H),8.21-8.19(m,2H),7.81(d,J＝8.0Hz,1H),7.54(s,1H),7.20(d,J＝9.2Hz,1H),6.75(d,J＝9.2Hz,1H),3.16(s,4H),2.44(s,4H),2.21(s,3H),1.49(s,6H)；LC-MS:[M+H] + :531.6。
EXAMPLE 13 Synthesis of Compound PR-01-081
Synthesis of 6- (2, 5-dichloropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (2):
the compound 2,4, 5-trichloropyrimidine (166.1 mg,0.90 mmol), 3-dimethyl-6- (pinacol borate) isoindol-1-one (200.0 mg,0.70 mmol) was dissolved in 1,4 dioxane (5 mL) and water (1 mL), and potassium carbonate (192.5 mg,1.39 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were added ]Palladium dichloride dichloromethane Complex (56.7 mg,0.07 mmol) then under N 2 The reaction was heated to 90 ℃ with stirring for 12 hours, LCMS detected complete reaction, the reaction was poured into water (10 mL), extracted with ethyl acetate (15 ml×3), the organic phases combined, washed twice with water (40 ml×2), once with 40mL brine, and concentrated to give the crude product. The crude product was subjected to column chromatography (ethyl acetate: petroleum ether=0-30%, ethyl acetate: petroleum ether=1:5, rf=0.3) to give a white solid (105 mg). LC-MS: [ M+H ]] + :307.9。
Synthesis of 6- (5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-081):
the compound 6- (2, 5-dichloropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (100 mg,0.32 mmol) and 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (88.1 mg,0.32 mmol) were dissolved in dioxane (5 mL), then palladium acetate (7.2 mg,0.032 mmol), cesium carbonate (208.5 mg,0.64 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (43.6 mg,0.07 mmol) were added to displace three times nitrogen and heated to 100℃for reaction for 12 hours. The reaction mixture was cooled to room temperature and concentrated in vacuo to remove most of the solvent. It was poured into water (5 mL) and extracted with ethyl acetate (30 mL). The organic layer was dried over magnesium sulfate, filtered and concentrated. The solid was prepared by basicity to give a white solid (12.5 mg). 1 H NMR(400MHz,DMSO)δ9.28(s,1H),8.80(s,1H),8.62(s,1H),7.97-8.01(m,2H),7.78(d,J＝7.6Hz,1H),7.53(d,J＝2.8Hz,1H),7.19(d,J＝8.8Hz,1H),6.72-6.75(m,1H),3.16–3.06(m,4H),2.44–2.32(m,4H),2.20(s,3H),1.49(s,6H)；LC-MS:[M+H] + :547.4。
EXAMPLE 14 Synthesis of Compound PR-01-037
Synthesis of methyl 2-bromo-5- (dibromomethyl) thiazole-4-carboxylate (2):
to a solution of carbon tetrachloride (35.0 mL) under nitrogen was added methyl 2-bromo-5-methylthiazole-4-carboxylate (5.0 g,21.18 mmol), N-bromosuccinimide (11.3 g,63.48 mmol), dibenzoyl peroxide (513.5 mg,2.12 mmol). The reaction mixture was heated to 80℃and stirred for 5 hours, after completion of the reaction, quenched with water (50.0 mL), extracted with dichloromethane (50 mL. Times.3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. Column chromatography (PE: ea=100:1) gave a pale yellow solid (3.3 g).
Synthesis of 2-bromo-5-formylthiazole-4-carboxylic acid methyl ester (3):
to a mixture of ethanol (300.0 mL) and water (100.0 mL) under nitrogen was added methyl 2-bromo-5- (dibromomethyl) thiazole-4-carboxylate (16.0 g,40.60 mmol) and silver nitrate (13.8 g,81.24 mmol). The system was heated to 80 ℃ and stirred for 3 hours, cooled and filtered after the reaction was complete. The mixture was extracted three times with dichloromethane (100 mL. Times.3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (PE: ea=10:1) gave a white solid (5.6 g). LC-MS: [ M+H ] ] + :249.9,252.0。
Synthesis of methyl 2-bromo-5- (hydroxymethyl) thiazole-4-carboxylate (4):
the compound methyl 2-bromo-5-formylthiazole-4-carboxylate (5.6 g,22.31 mmol) was dissolved in methanol (100.0 mL), the system was cooled to 0deg.C, sodium borohydride (1.3 g,34.36 mmol) was slowly added, and the mixture was stirred at room temperature for 1 hour until the reaction was complete, and the temperature was loweredThe reaction was quenched by addition of saturated aqueous ammonium chloride (50.0 mL), extracted three times with ethyl acetate (50 mL. Times.3), the combined organic phases were washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (PE: ea=4:1) gave a white solid (3.7 g). LC-MS: [ M+H ]] + :252.0,254.0。
Synthesis of methyl 5- (hydroxymethyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) thiazole-4-carboxylate (5):
to a suspension of methyl 2-bromo-5- (hydroxymethyl) thiazole-4-carboxylate (1.3 g,5.14 mmol), 5-methyl-2- (methylthio) -4- (trimethylstannyl) pyrimidine (1.4 g,4.62 mmol) in dioxane (40 mL) was added bis (triphenylphosphine) palladium dichloride (356.6 mg,0.51 mmol) and cuprous iodide (194.3 mg,1.02 mmol). The reaction system was stirred at 110℃for 2 hours under nitrogen protection. After the reaction was completed, an aqueous potassium fluoride solution (50.0 mL) was added to quench the reaction, the reaction was extracted three times with methylene chloride (50 ml×3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. Column chromatography (PE: ea=2:1) gave a brown oily liquid (600.0 mg). LC-MS: [ M+H ] ] + :312.0. Synthesis of methyl 2- (5-methyl-2- (methylsulfanyl) pyrimidin-4-yl) -5- (((methylsulfonyl) oxy) methyl) thiazole-4-carboxylate (6):
the compound methyl 5- (hydroxymethyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) thiazole-4-carboxylate (1.0 g,3.22 mmol) and triethylamine (649.6 mg,6.42 mmol) were dissolved in dichloromethane (10.0 mL). The system was cooled to 0℃and methanesulfonyl chloride (551.8 mg,4.82 mmol) was slowly added dropwise to the system. After the completion of the dropwise addition, the reaction was continued with stirring at 0℃for 2 hours. After the reaction was completed, the reaction was quenched by addition of saturated aqueous ammonium chloride (15.0 mL), extracted three times with ethyl acetate (15 mL. Times.3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (DCM: meoh=10:1) afforded a brown oily liquid (560.0 mg). LC-MS: [ M+H ]] + :389.9。
Synthesis of methyl 5- (((2, 4-dimethoxybenzyl) amino) methyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) thiazole-4-carboxylate (7):
the compound 2, 4-dimethoxybenzylamine (2.0 g,11.96 mmol) and triethylamine (1.2 g,11.86 mmol) were dissolved in acetonitrile (50.0 mL). The system was cooled to 0deg.C, and methyl 2- (5-methyl-2- (methylsulfanyl) pyrimidin-4-yl) -5- (((methylsulfonyl) oxy) methyl) thiazole-4-carboxylate (930.0 mg,2.39 mmol) was slowly added dropwise to the system. After the completion of the dropwise addition, the reaction was continued with stirring at 0℃for 2 hours. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure and isolated by column chromatography (DCM: meoh=10:1) to give a yellow solid (540.0 mg). LC-MS: [ M+H ] ] + :461.1。
Synthesis of 5- (((2, 4-dimethoxybenzyl) amino) methyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) thiazole-4-carboxylic acid (8):
the compound 5- (((2, 4-dimethoxybenzyl) amino) methyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) thiazole-4-carboxylic acid methyl ester (575.0 mg,1.25 mmol), lithium hydroxide monohydrate (104.9 mg,2.50 mmol) was dissolved in a mixed solution of tetrahydrofuran (2.0 mL) and water (4.0 mL). The reaction was stirred at room temperature for 2 hours. After the reaction is completed, 2N hydrochloric acid is added to adjust the pH of the solution to 3-4, and the solvent is removed by spin-drying under reduced pressure to obtain a brown yellow oily product without further purification. LC-MS: [ M+H ]] + :447.1。
Synthesis of 5- (2, 4-dimethoxybenzyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrolo [3,4-d ] thiazol-4-one (9):
the above crude product and N, N-diisopropylethylamine (484.7 mg,3.75 mmol) were dissolved in N, N-dimethylformamide (60.0 mL), the system was cooled to 0℃and 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (570.4 mg,1.50 mmol) was slowly added to the system. After the addition was completed, the temperature was raised to room temperature, and the reaction was stirred for 1 hour. After the reaction was completed, the reaction was quenched with water, extracted three times with ethyl acetate (50 ml×3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (DCM: meoh=10:1) afforded a yellow oil (125.0 mg). LC-MS: [ M+H ] ] + :429.1。
Synthesis of 5- (2, 4-dimethoxybenzyl) -2- (5-methyl-2- (methylsulfonyl) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrolo [3,4-d ] thiazol-4-one (10):
5- (2, 4-Dimethoxybenzyl) -2- (5-methyl-2- (methylthio) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrole [3,4-d]Thiazol-4-one (200.0 mg,0.47 mmol) was dissolved in dichloromethane (4.0 mL), the system was cooled to 0 ℃ and m-chloroperoxybenzoic acid (161.1 mg,0.93 mmol) was slowly added to the system. After the addition was completed, the reaction was stirred at 0℃for 2 hours. After the reaction was completed, the reaction was quenched by adding an aqueous sodium bicarbonate solution, the organic phase was washed three times with sodium bicarbonate, then with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (DCM: meoh=10:1) afforded a yellow solid (85.0 mg). LC-MS: [ M+H ]] + :461.1。
Synthesis of 5- (2, 4-dimethoxybenzyl) -2- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrolo [3,4-d ] thiazol-4-one (11):
5- (2, 4-Dimethoxybenzyl) -2- (5-methyl-2- (methylsulfonyl) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrole [3,4-d]Thiazol-4-one (30.0 mg,0.07 mmol), N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) carboxamide (common int-3) (23.7 mg,0.08 mmol), potassium carbonate (18.0 mg,0.13 mmol) were dissolved in N, N-dimethylformamide (1.0 mL). The reaction system was stirred at 90℃for 16 hours under nitrogen protection. After the reaction was completed, the reaction was quenched by addition of saturated aqueous ammonium chloride (5.0 mL), extracted three times with methylene chloride (10 mL. Times.3), the organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (DCM: meoh=10:1) afforded a yellow oil (4.0 mg). LC-MS: [ M+H ] ] + :656.2。
Synthesis of 5-methyl-2- (methylthio) -4- (trimethylstannyl) pyrimidine (13):
to a suspension of 4-chloro-5-methyl-2- (methylthio) pyrimidine (4.5 g,25.77 mmol) and hexamethylditin (16.9 g,51.54 mmol) in 1, 4-dioxane (50 mL) was added tetrakis (triphenylphosphine) palladium (1.5 g,1.29 mmol). The reaction system was stirred at 80℃for 12 hours under nitrogen protection. After the reaction was completed, the reaction was quenched by adding an aqueous potassium fluoride solution (50.0 mL), extracted with methylene chloride (50 mL. Times.3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to giveCrude product. Column chromatography (PE: ea=2:1) gave a colourless oil (3.0 g). LC-MS: [ M+H ]] + :305.1。
Synthesis of 2- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrolo [3,4-d ] thiazol-4-one (PR-01-037):
5- (2, 4-Dimethoxybenzyl) -2- (5-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -5, 6-dihydro-4H-pyrrolo [3,4-d]Thiazol-4-one (13.0 mg,0.02 mmol) was dissolved in trifluoroacetic acid (0.5 mL), triethylsilane (0.1 mL), dichloromethane (0.5 mL). The reaction system was stirred at 50℃for 12 hours under nitrogen protection. After the reaction is completed, the mixture is directly concentrated to dryness. The crude product was purified by preparative liquid chromatography to give a yellow solid (4.0 mg). 1 H NMR(400MHz,CD 3 OD)δ8.53(s,1H),8.09(s,1H),7.24(d,J＝9.2Hz 1H),6.74(d,J＝9.2Hz,1H),4.58(s,2H),3.42(s,4H),3.04(s,4H),2.65(s,6H)；LC-MS:[M+H] + :506.1。
EXAMPLE 15 Synthesis of Compound PR-01-091
Synthesis of tert-butyl 1-oxo-3, 5-dihydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate (2):
p-toluenesulfonylmethyloisonitrile (586.0 mg,3.00 mmol) was dissolved in tetrahydrofuran (10 mL), and 1, 8-diazabicyclo [5.4.0 ] was added with stirring at room temperature]Undec-7-ene (457.0 mg,3.00 mmol) was added to the solution, followed by stirring for 15 minutes, and then 2-oxo-2, 5-dihydro-1H-pyrrole-1-carboxylic acid tert-butyl ester (500.0 mg,2.73 mmol) was reacted under stirring at room temperature for 5 hours. After completion of the reaction, water (20 mL) was added to the system, followed by extraction with ethyl acetate (30 ml×3), and the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by column chromatography (dichloromethane: methanol=50:1-10:1, dichloromethane: methanol=10:1, rf=0.3) to give a brown solid (260.0 mg). LCMS [ M+H ]] + :223.2。
Synthesis of tert-butyl 5- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrole [3,4-c ] pyrrole-2 (1H) -carboxylate (3):
the compound 1-oxo-3, 5-dihydropyrrolo [3,4-c]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (80.0 mg,0.36 mmol) was dissolved in N, N-dimethylformamide (5 mL) and 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-2) (140.0 mg,0.36 mmol) and cesium carbonate (235.0 mg,0.72 mmol) were added under nitrogen. The system was heated to 100℃and reacted for 1 hour. The reaction was added with water (10 mL), extracted with ethyl acetate (30 ml×3), and concentrated to give crude product which was purified by preparative TLC (dichloromethane: alcohol=20:1, rf=0.4) to give yellow solid (40.0 mg). LCMS [ M+H ] ] + :574.6。
Synthesis of 5- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 5-dihydropyrrol [3,4-c ] pyrrol-1 (2H) -one (PR-01-091):
the compound 5- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrol [3,4-c]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (40.0 mg,0.07 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.2 mL) was added and the reaction was stirred at room temperature for 1 hour, the reaction solution was concentrated to dryness, and a white solid (23.5 mg) was obtained by preparative purification. 1 H NMR(400MHz,DMSO-d6)δ9.16(s,1H),8.48(d,J＝5.6Hz,1H),8.04(s,1H),7.93(s,1H),7.57(s,1H),7.35(d,J＝2.8Hz,1H),7.23(d,J＝6.0Hz,2H),6.80(dd,J＝9.2,2.8Hz,1H),4.23(s,2H),3.17(s,4H),2.46(s,4H),2.22(s,3H)；LCMS:[M+H] + :474.1。
EXAMPLE 16 Synthesis of Compounds PR-01-098
Synthesis of tert-butyl 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate (2):
the compound 1-oxo-3, 5-dihydropyrrolo [3,4-c]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester 1 (220.0 mg,0.99 mmol), 2, 4-dichloro-5-fluoropyrimidine (165.0 mg,0.99 mmol) were dissolved in acetonitrile (10 mL) and potassium carbonate (410) was added under nitrogen.0mg,2.97 mmol). The system was heated to 80℃and reacted for 12 hours. After the completion of the reaction, the reaction mixture was concentrated to dryness, water (10 mL) was added to the reaction system, extracted with ethyl acetate (20 ml×3), and the crude product was concentrated to give a pale yellow solid (104.0 mg) by preparative TLC (dichloromethane: methanol=20:1, rf=0.6). LCMS: [ M ] t Bu+H] + :296.9。
Synthesis of tert-butyl 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate (3):
the compound 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrolo [3,4-c]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (151.0 mg,0.43 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (117.0 mg,0.43 mmol), palladium acetate (9.0 mg,0.04 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (56.0 mg,0.09 mmol) were dissolved in 1, 4-dioxane (2 mL), and cesium carbonate (277.0 mg,0.85 mmol) was added under nitrogen. After the reaction was raised to 100 ℃ and stirred for 6 hours LCMS detected completion of the reaction. Water was added to the reaction solution and extracted with dichloromethane (20 ml×3), and the organic phases were combined, dried and concentrated to give a crude product which was purified by column chromatography (dichloromethane: methanol=20:1, rf=0.3) to give a brown yellow solid (85.0 mg). LCMS [ M+H ]] + :592.3。
Synthesis of 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 5-dihydropyrrol [3,4-c ] pyrrol-1 (2H) -one (PR-01-098):
the compound 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-oxo-3, 5-dihydropyrrolo [3,4-c ]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (85.0 mg,0.14 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.2 mL) was added. The reaction was stirred at room temperature for 1 hour, and the reaction solution was concentrated to dryness to give a white solid (29.5 mg) by preparative purification. 1 H NMR(400MHz,DMSO-d6)δ9.27(s,1H),8.66(d,J＝4.0Hz,1H),8.09(s,1H),7.72(s,1H),7.44(s,1H),7.34(d,J＝2.8Hz,1H),7.23(d,J＝8.0Hz,1H),6.79(dd,J＝9.2,2.8Hz,1H),4.24(s,2H),3.19–3.13(m,4H),2.47–2.42(m,4H),2.22(s,3H)；LCMS:[M+H] + :492.2。
EXAMPLE 17 Synthesis of Compounds DP-01-130
Synthesis of 1- (4-bromo-1H-pyrrol-2-yl) -N- (2, 4-dimethoxybenzyl) methylamine (2):
the compound 4-bromo-1H-pyrrole-2-carbaldehyde (2.0 g,11.50 mmol) and 2, 4-dimethoxybenzylamine (1.92 g,11.48 mmol) were dissolved in methanol (20 mL), stirred at room temperature for 3 hours, cooled to 0deg.C, sodium borohydride (0.43 g,11.37 mmol) was added in portions, and stirred at room temperature for 1 hour, and LCMS monitored the starting material was reacted. The reaction mixture was quenched with water (40 mL), and the methanol was removed by swirling, followed by extraction with ethyl acetate (30 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and rotary distilled to give crude product. The crude product was chromatographed on a column of silica gel (methanol: dichloromethane=0-3%; dichloromethane/methanol=10/1, rf=0.6) to give a yellow oil (3.2 g). LC-MS: [ M+H ]] + :325.0,327.0。
Synthesis of 6-bromo-2- (2, 4-dimethoxybenzyl) -1, 2-dihydro-3H-pyrrolo [1,2-c ] imidazol-3-one (3):
the compound 1- (4-bromo-1H-pyrrol-2-yl) -N- (2, 4-dimethoxybenzyl) methylamine (500.0 mg,1.54 mmol) was dissolved in THF (8 mL), nitrogen was replaced three times, cooled to 0deg.C, N' -carbonyldiimidazole (374.0 mg,2.31 mmol) was added in portions, and the mixture was returned to room temperature and stirred for 1 hour. Cooling to 0 ℃, 60% sodium hydride (123.0 mg,3.08 mmol) was added in portions, the temperature was returned to room temperature and stirring was continued for 2 hours, LCMS monitored the starting material had reacted. The reaction mixture was quenched with saturated ammonium chloride solution (20 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-20%, petroleum ether/ethyl acetate=1:5, rf=0.4) to give a bright yellow solid (450.0 mg). LC-MS: [ M+H ] ] + :351.1,353.1。
Synthesis of 2- (2, 4-dimethoxybenzyl) -6-boronic acid pinacol ester-1, 2-dihydro-3H-pyrrolo [1,2-c ] imidazol-3-one (4):
the compound 6-bromo-2- (2, 4-dimethoxybenzyl) -12-dihydro-3H-pyrrolo [1,2-c]Imidazol-3-one (400.0 mg,1.14 mmol), pinacol biborate (433.8 mg,1.71 mmol), potassium acetate (223.6 mg,2.28 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (82.7 mg,0.113 mmol) was dissolved in 1, 4-dioxane (10 mL), heated to 80℃under nitrogen for 4 hours, and LCMS monitored the starting material reacted. The reaction solution was directly used for the next reaction. LC-MS: [ M+H ]] + :399.0。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -2- (2, 4-dimethoxybenzyl) -1, 2-dihydro-3H-pyrrolo [1,2-c ] imidazol-3-one (5):
the compound 2- (2, 4-dimethoxy benzyl) -6-boric acid pinacol ester-1, 2-dihydro-3H-pyrrolo [1,2-c]Imidazol-3-one (400.0 mg,1.00 mmol), 2, 4-dichloro-5-fluoropyrimidine (167.7 mg,1.00 mmol), potassium carbonate (416.4 mg,3.01 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (72.9 mg,0.10 mmol) was dissolved in 1, 4-dioxane (10 mL) and water (3 mL), nitrogen was replaced three times, and the temperature was raised to 80℃and stirred for 4 hours. LCMS monitored complete reaction of the starting material, quench with water (20 mL), separate the liquid, collect the organic phase and extract the aqueous phase with dichloromethane (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-25%; petroleum ether/ethyl acetate=4/1, rf=0.3) to give a white solid (72.0 mg). LC-MS: [ M+H ] ] + :403.1。
Synthesis of 2- (2, 4-dimethoxybenzyl) -6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1, 2-dihydro-3H-pyrrolo [1,2-c ] imidazol-3-one (6):
the compound 6- (2-chloro-5-fluoropyrimidin-4-yl) -2- (2, 4-dimethoxybenzyl) -1, 2-dihydro-3H-pyrrolo [1,2-c]Imidazol-3-one (60.0 mg,0.15 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (41.0 mg,0.15 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (18.6 mg,0.03 mmol), palladium acetate (4.5 mg,0.02 mmol) and cesium carbonate (145.6 mg,0.45 mmol) were dissolved in dioxane (5 mL), nitrogen was replaced, heated to 90℃and stirred for 6 hours, and LCMS monitored the starting material to have reacted. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The combined organic phases were washed with saturated brine (10 mL), anhydrous sodium sulfateDrying, filtering and concentrating to obtain crude product. Purification by silica gel column chromatography (methanol: dichloromethane=0-3%; dichloromethane/methanol=20:1, rf=0.4) afforded a yellow solid (90.0 mg). LC-MS: [ M+H ]] + :642.3。
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1, 2-dihydro-3H-pyrrolo [1,2-c ] imidazol-3-one (DP-01-130):
The compound 2- (2, 4-dimethoxybenzyl) -6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1, 2-dihydro-3H-pyrrolo [1,2-c]Imidazol-3-one (90.0 mg,0.14 mmol) was dissolved in dichloromethane (2.5 mL), trifluoroacetic acid (5 mL) and triethylsilane (2 mL) were added to the above solution, and the mixture was stirred for 16 hours at 50℃and LCMS monitored that the starting material had reacted. The reaction solution was dried by spin-drying, followed by preparation and purification to give a white solid (17.0 mg). 1 H NMR(400MHz,CDCl 3 )δ8.32(s,1H),8.29(s,1H),7.93(s,1H),7.37(s,1H),7.15(d,J＝8.8Hz,1H),6.83(s,1H),6.53(d,J＝8.8Hz,1H),5.92(s,1H),4.51(s,2H),3.33(s,4H),2.67(s,4H),2.41(s,3H)；LC-MS:[M+H] + :492.3。
EXAMPLE 18 Synthesis of Compounds PR-01-096
Synthesis of 5-acetyl-2-bromo-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (2):
the compound 5-acetyl-6, 6-dimethyl-5, 6-dihydro-4H-thiophene [2,3-c]Pyrrole-4-one (450.0 mg,2.15 mmol) was dissolved in a mixed solution of acetic acid (12.0 mL) and water (10.0 mL), the system was cooled to 0℃and bromine (377.9 mg,2.36 mmol) was slowly added dropwise to the system, and after the dropwise addition, the reaction was continued at 0℃for 1 hour. After the reaction was completed, water (15 mL) was added for dilution, extraction with ethyl acetate (15 mL. Times.3), and the organic phases were combined, washed with a saturated aqueous sodium sulfite solution (15 mL) and a saturated brine (10 mL) in this order, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Column chromatography (DCM: meoh=10:1) afforded a white solid (583.2 mg). LC-MS: [ M+H ] ] + :288.0,290.0。
Synthesis of 5-acetyl-6, 6-dimethyl-2-boronic acid pinacol ester-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (3):
the compound 5-acetyl-2-bromo-6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (100.0 mg,0.35 mmol), potassium acetate (102.4 mg,1.04 mmol), 1-bis (diphenylphosphine) ferrocene palladium dichloride (25.4 mg,0.04 mmol), pinacol biborate (114.8 mg,0.45 mmol) were dissolved in 1, 4-dioxane (3 mL). The system was evacuated to replace nitrogen three times, heated to 80 ℃ and reacted for 1 hour, and after the reaction was completed, the reaction was cooled to room temperature. The system is a brownish-black turbid solution, and is directly used for the next reaction without further treatment. LC-MS: [ M+H ]] + :336.2。
Synthesis of 5-acetyl-2- (5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (4):
the compound 5-acetyl-6, 6-dimethyl-2-boronic acid pinacol ester-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (81.5 mg,0.24 mmol), 4, 5-dichloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (common int-5) (123.0 mg,0.29 mmol), 1-bis (diphenylphosphine) ferrocene palladium dichloride (17.8 mg,0.02 mmol) and potassium phosphate (154.7 mg,0.73 mmol) were dissolved in a solution of 1, 4-dioxane (2.5 mL), the system was evacuated to replace nitrogen three times, and after heating to 90℃for 2 hours the reaction was cooled to room temperature. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL. Times.3), the combined organic phases were successively washed with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was isolated as a pale yellow solid (72.1 mg) by preparative separation. LC-MS: [ M+H ] ] + :595.1。
Synthesis of 2- (5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (PR-01-096):
the compound 5-acetyl-2- (5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5,6-dihydro-4H-thieno [2,3-c ]]Pyrrole-4-one (100.0 mg,0.17 mmol) and sodium hydroxide (20.2 mg,0.51 mmol) were dissolved in a mixed solution of methanol (4.0 mL) and water (4.0 mL), and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, water (10 mL) was added for dilution, the aqueous phase was extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined and washed successively with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was isolated as a yellow solid (2.5 mg) by preparation. 1 H NMR(400MHz,Methanol-d4)δ8.50(s,1H),8.30(s,1H),7.90(d,J＝2.8Hz,1H),7.27(dd,J＝9.2,1.2Hz,1H),6.83(dd,J＝9.2,2.8Hz,1H),3.89(m,2H),3.61(m,2H),3.29(m,2H),3.17(m,2H),2.97(s,3H),1.63(s,6H)；LC-MS：[M+H] + :553.1。
EXAMPLE 19 Synthesis of Compound PR-01-097
Synthesis of 5-acetyl-2- (2-chloro-5-fluoropyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (2):
the compound 5-acetyl-6, 6-dimethyl-2-boronic acid pinacol ester-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (116.3 mg,0.35 mmol), 2, 4-dichloro-5-fluoropyrimidine (69.5 mg,0.42 mmol), 1-bis (diphenylphosphino) ferrocene palladium dichloride (25.4 mg,0.04 mmol), potassium phosphate (221.0 mg,1.04 mmol) were dissolved in 1, 4-dioxane (4.0 mL) and water (0.4 mL), and the system was evacuated to replace nitrogen three times. Heating to 90 ℃ for reaction for 2 hours, and cooling the reaction liquid to room temperature after the reaction is complete. The reaction was diluted with water (10 mL), extracted with ethyl acetate (10 mL. Times.3), and the organic phases were combined and washed sequentially with water (10 mL) and saturated aqueous sodium chloride (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. Column chromatography (DCM: meoh=10:1) afforded a white solid (91.2 mg). LC-MS: [ M+H ] ] + :340.0。
Synthesis of 5-acetyl-2- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (3):
the compound 5-acetyl-2- (2-chloro-5-fluoropyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (92.0 mg,0.27 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (82.0 mg,0.30 mmol), palladium acetate (9.0 mg,0.04 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (33.7 mg,0.05 mmol) and cesium carbonate (176.6 mg,0.54 mmol) were dissolved in 1, 4-dioxane (5.0 mL), and the system was evacuated to replace nitrogen three times and heated to 100℃for reaction for 16 hours. After the reaction was complete, the reaction was cooled to room temperature. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL. Times.3), the combined organic phases were successively washed with water (10 mL) and saturated aqueous sodium chloride solution (10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. Chromatography on silica gel (DCM: meoh=10:1) afforded a yellow solid (98.4 mg). LC-MS: [ M+H ]] + :579.3。
Synthesis of 2- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (PR-01-097):
The compound 5-acetyl-2- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (110.0 mg,0.19 mmol) and sodium hydroxide (22.8 mg,0.57 mmol) were dissolved in a mixed solution of methanol (4.0 mL) and water (4.0 mL), and the reaction was stirred at room temperature for 2 hours. After the reaction was complete, water (10 mL) was added, extracted with ethyl acetate (10 mL. Times.3), and the combined organic phases were washed successively with water (10 mL) and saturated aqueous sodium chloride (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was isolated as a yellow solid (34.4 mg) by preparative separation. 1 H NMR(400MHz,DMSO-d6)δ9.03(s,1H),8.68(s,1H),8.63(d,J＝3.2Hz,1H),7.71(d,J＝1.6Hz,1H),7.55(d,J＝2.8Hz,1H),7.20(d,J＝8.0Hz,1H),6.74(dd,J＝8.8,2.8Hz,1H),3.23–3.15(m,4H),2.47–2.40(m,4H),2.21(s,3H),1.54(s,6H)；LC-MS:[M+H] + :537.2。
EXAMPLE 20 Synthesis of Compounds PR-01-107
Synthesis of 5-bromo-1-methyl-1, 2-dihydro-3H-indazol-3-one (2):
methyl 5-bromo-2-fluorobenzoate (1.0 g,4.29 mmol) and methylhydrazine (0.2 g,4.34 mmol) were dissolved in N, N-dimethylacetamide (10 mL), and after heating to 150℃in a closed tube for 8 hours, the reaction mixture was cooled, diluted with ethyl acetate (50 mL), washed three times with saturated brine, and the organic phases were combined and dried over anhydrous sodium sulfate, and the organic phase was concentrated in vacuo and column chromatographed (petroleum ether/ethyl acetate=1/10) to give a colorless liquid (580.0 mg). LC-MS: [ M+H ] ] + :226.9,228.9。
Synthesis of 1-methyl-5-boronic acid pinacol ester-1, 2-dihydro-3H-indazol-3-one (3):
the compound 5-bromo-1-methyl-1, 2-dihydro-3H-indazol-3-one (250.0 mg,1.10 mmol), pinacol biborate (419.3 mg,1.65 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (89.9 mg,0.11 mmol) and potassium acetate (324.2 mg,3.30 mmol) were dissolved in 1, 4-dioxane (2 mL) and heated to 80℃under nitrogen for 3 hours. Ethyl acetate (10 mL) was added to the reaction mixture to dilute it, and the mixture was filtered. The filtrate was washed three times with water and saturated brine, and the organic phases were combined and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (petroleum ether/ethyl acetate=1/8) to give the product as a white solid (210.0 mg). LC-MS: [ M+H ]] + :275.1。
Synthesis of 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-methyl-1, 2-dihydro-3H-indazol-3-one (4):
the compound 1-methyl-5-boronic acid pinacol ester-1, 2-dihydro-3H-indazol-3-one (200.0 mg,0.73 mmol), 2, 4-dichloro-5-fluoropyrimidine (182.7 mg,1.09 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (59.6 mg,0.07 mmol) and potassium phosphate (464.6 mg,2.19 mmol) were dissolved in 1, 4-dioxane (2 mL) and water (2 mL) and heated to 80℃under nitrogen for 2 hours. Ethyl acetate (10 mL) was added to the reaction mixture to dilute the mixture, and the resulting mixture was separated by extraction. The organic phases were combined and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (petroleum ether/ethyl acetate=1/3) to give a white solid product Substance (130.0 mg). LC-MS: [ M+H ]] + :279.0。
Synthesis of 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-methyl-1, 2-dihydro-3H-indazol-3-one (PR-01-107):
the compound 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-methyl-1, 2-dihydro-3H-indazol-3-one (100.0 mg,0.36 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (59.5 mg,0.22 mmol), cesium carbonate (223.0 mg,0.69 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (44.7 mg,0.07 mmol) and palladium acetate (11.2 mg,0.05 mmol) were dissolved in 1, 4-dioxane (2 mL) and heated to 100℃under nitrogen for reaction for 4 hours. Ethyl acetate (10 mL) was added to the reaction mixture to dilute the mixture, and the resulting mixture was separated by extraction. The organic phases were combined and dried over anhydrous sodium sulfate, concentrated and purified by reverse phase preparation to give the product as a pale yellow solid (96.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.90(s,1H),8.48(d,J＝2.8Hz,1H),7.64(d,J＝8.8Hz,1H),7.46–7.40(m,2H),7.11–6.99(m,2H),6.91(d,J＝2.8Hz,1H),6.66(dd,J＝6.4,2.8Hz,1H),3.99(s,3H),2.94–2.87(m,4H),2.39–2.31(m,4H),2.20(s,3H)；LC-MS:[M+H] + :518.3。
EXAMPLE 21 Synthesis of Compounds PR-01-083, PR-01-083A, PR-01-083B
Synthesis of 6-bromo-1-oxoisoindole-2-carboxylic acid tert-butyl ester (2):
the compound 6-bromo-1-isoindolinone (1.0 g,4.72 mmol) was dissolved in dichloromethane (20 mL), di-tert-butyl dicarbonate (1.5 g,6.87 mmol) and 4-dimethylaminopyridine (288.3 mg,2.36 mmol) were added under ice-bath and stirred at room temperature for 6 hours. The reaction solution was poured into water (20 mL), then extracted with dichloromethane (40 ml×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by column chromatography (petroleum ether: ethyl acetate=10:1, rf=0.5) to give a white solid (1.2 g). LC-MS: [ M+H ] ] + :314.0。
Synthesis of 5-bromo-1-methyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (3):
the compound tert-butyl 6-bromo-1-oxoisoindole-2-carboxylate (500.0 mg,1.60 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), the reaction was purged three times with nitrogen and cooled to-78℃and 1M lithium bis trimethylsilylamide (1.9 mL,1.92 mmol) was added dropwise under protective conditions, and methyl iodide (454.9 mg,3.20 mmol) was added after 1 hour of reaction at constant temperature. The reaction solution was gradually warmed to room temperature and stirred for 2 hours, and after completion of the reaction, the reaction solution was poured into an aqueous ammonium chloride solution (20 mL), then extracted with ethyl acetate (40 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by chromatography (petroleum ether: ethyl acetate=10:1, rf=0.6) to give a pale yellow oil (250.0 mg). LC-MS: [ M ] t Bu+H] + :270.0,272.0。
Synthesis of 1-methyl-3-oxo-5-boronic acid pinacol ester isoindole-2-carboxylic acid tert-butyl ester (4):
the compound tert-butyl 5-bromo-1-methyl-3-oxoisoindole-2-carboxylate (250.0 mg,0.77 mmol) was dissolved in 1, 4-dioxane (5 mL) and 1,1' -bis (diphenylphosphine) ferrocene was added sequentially]Palladium dichloride (56.1 mg,0.08 mmol), pinacol biborate (253.0 mg,1.00 mmol) and potassium acetate (225.7 mg,2.30 mmol) were replaced 3 times with nitrogen. The reaction mixture was heated to 80℃and reacted for 2 hours. After the reaction was completed, the reaction solution was washed with water (15 mL) and ethyl acetate (20 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness, purified by chromatography column (petroleum ether: ethyl acetate=30:1, rf=0.5), and concentrated to give a yellow oil (242.0 mg). LC-MS [ M-Boc+H ] ] - :272.0。
Synthesis of 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-methyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (5):
the compound 1-methyl-3-oxo-5-boronic acid pinacol ester isoindole-2-carboxylic acid tert-butyl ester (240.0 mg,0.64 mmol), 1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (47.1 mg,0.06 mmol) was dissolved in 1, 4-dioxane (6 mL) and water (2 mL), and potassium phosphate (409.4 mg,1.93 mmol) was added under nitrogen protection, and the reaction mixture was heated to 100℃for 1 hour. After the completion of the reaction, the reaction solution was poured into an aqueous solution (30 mL), followed by extraction with ethyl acetate (30 mL. Times.3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered,concentrated to dryness and purified by column chromatography (petroleum ether: ethyl acetate=5:1, rf=0.5) to give a yellow solid (94.0 mg). LC-MS: [ M ] t Bu+H] + :322.0。
Synthesis of tert-butyl 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-methyl-3-oxoisoindole-2-carboxylate (6):
the compound tert-butyl 5- (2-chloro-5-fluoropyrimidin-4-yl) -1-methyl-3-oxoisoindole-2-carboxylate (80.0 mg,0.21 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (58.3 mg,0.21 mmol), palladium acetate (6.7 mg,0.03 mmol) and 1,1 '-binaphthyl-2, 2' -diphenylphosphine (26.4 mg,0.04 mmol) were dissolved in anhydrous 1, 4-dioxane (3 mL), cesium carbonate (138.0 mg,0.42 mmol) was added under nitrogen protection and the reaction mixture was reacted at 90℃for 5 hours. After completion of the reaction, water was added to dilute (20 mL), and the mixture was extracted with methylene chloride (30 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness and purified by preparative TLC (dichloromethane: methanol=20:1, rf=0.4) to give a yellow solid (95.0 mg). LC-MS: [ M+H ] ] + :617.2。
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3-methylisoindol-1-one (PR-01-083):
the compound 5- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -1-methyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (20.0 mg,0.03 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.2 mL) was added and stirred at room temperature for 1 hour, the reaction mixture was concentrated to dryness, and a white solid (2.5 mg) was isolated by preparation. 1 H NMR(400MHz,CDCl 3 )δ8.57(s,1H),8.44(s,1H),8.38(d,J＝7.6Hz,1H),8.31(s,1H),7.57(d,J＝8.0Hz,1H),7.46(s,1H),7.16(d,J＝9.2Hz,1H),6.55(d,J＝8.4Hz,1H),6.20(s,1H),4.79(d,J＝6.4Hz,1H),3.28(s,4H),2.60(s,4H),2.38(s,3H),1.56(s,3H)；LC-MS:[M+H] + :517.3。
The racemate PR-01-083 was purified by SFC (column: chiralPak IB, 250X 30mm I.D.,5 μm; mobile phase: A phase: carbon dioxide, B phase: [ MEOH+0.1%7mol/L NH.) 3 ]The method comprises the steps of carrying out a first treatment on the surface of the Gradient: b%:20% -20%,10.8min;120 min) to obtainTo white solids PR-01-083A and PR-01-083B.
PR-01-083A:
Chiral analysis conditions: chromatographic column: CHIRALPAK IB 100 x 4.6mm 5 μm; mobile phase: phase A: carbon dioxide; and B phase: meOH (0.05% DEA v/v); gradient: b%:20% -20%; retention time: 3.059min; ee% = 99.7%.
1 H NMR(400MHz,MeOD)δ8.48(s,2H),8.31(d,J＝8.0Hz,1H),8.20(s,1H),7.72(d,J＝8.0Hz,1H),7.24(d,J＝9.2Hz,1H),6.76(d,J＝8.8Hz,1H),4.81–4.76(m,1H),3.47(s,6H),3.31(s,2H),2.99(s,3H),1.52(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :517.2。
PR-01-083B:
Chiral analysis conditions: chromatographic column: CHIRALPAK IB 100 x 4.6mm 5 μm; mobile phase: phase A: carbon dioxide; and B phase: meOH (0.05% DEA v/v); gradient: b%:20% -20%; retention time: 3.692min; ee% = 99.1%.
1 H NMR(400MHz,MeOD)δ8.51(s,1H),8.46(s,1H),8.37(d,J＝8.0Hz,1H),8.13(s,1H),7.74(d,J＝8.4Hz,1H),7.19(d,J＝8.8Hz,1H),6.69(d,J＝9.2Hz,1H),4.81(q,J＝6.4Hz,1H),3.24(s,4H),2.61(s,4H),2.35(s,3H),1.52(d,J＝6.8Hz,3H)；LC-MS:[M+H] + :517.2。
EXAMPLE 22 Synthesis of Compounds PR-01-104
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (2, 2-trifluoroethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-104):
6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (150.0 mg,0.51 mmol), 5- (4-methylpiperazin-1-yl) -2- (2, 2-trifluoroethoxy) aniline (common int-6) (223.1 mg,0.77 mmol), palladium acetate (15.7 mg,0.07 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (64.0 mg,0.10 mmol) and cesium carbonate (502.6 mg,1.54 mmol) were suspended in 1, 4-dioxane (2 mL) and reacted under nitrogen protection at 100℃for 3 hours, the reaction mixtureCooling and filtering. The crude brown solid obtained after concentration of the filtrate was purified by preparation to give a yellow solid (59.8 mg). 1 H NMR(400MHz,DMSO-d6)δ8.83(s,1H),8.65(d,J＝3.6Hz,1H),8.28(s,1H),8.24–8.16(m,2H),7.87–7.77(m,2H),7.08(d,J＝8.8Hz,1H),6.66–6.62(m,1H),8.24–8.16(m,2H),3.09–3.07(m,4H),2.49–2.43(m,4H),2.23(s,3H),1.50(s,6H)；LC-MS:[M+H] + :545.2。
EXAMPLE 23 Synthesis of Compounds PR-01-121
Synthesis of 4-bromo-1- (difluoromethoxy) -2-nitrobenzene (2):
the compound 4-bromo-2-nitrophenol (1.0 g,4.59 mmol) was dissolved in anhydrous tetrahydrofuran (20.0 mL), replaced 3 times with nitrogen, 60% sodium hydride (1.8 g,45.87 mmol) was added under ice-bath conditions and stirred for 0.5 hours. Bromine difluoro methyl phosphonic acid diethyl ester (2.5 g,9.36 mmol) was slowly added dropwise and the reaction was allowed to warm to room temperature for 2 hours, the reaction solution was poured into ice water (20.0 mL), then extracted with ethyl acetate (30.0 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=20:1-5:1) to give a pale yellow oil (897.0 mg). 1 H NMR(400MHz,DMSO)δ8.34(s,1H),8.00(d,J＝8.8Hz,1H),7.50(d,J＝8.8Hz,1H),7.37(t,J＝72.0Hz,1H)。
Synthesis of 1- (4- (difluoromethoxy) -3-nitrophenyl) -4-methylpiperazine (3):
the compound 4-bromo-1- (difluoromethoxy) -2-nitrobenzene (550.0 mg,2.05 mmol) was dissolved in anhydrous toluene (6.0 mL) and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (121.5 mg,0.21 mmol) and tris (dibenzylideneacetone) dipalladium (94.0 mg,0.10 mmol) were added sequentially. Cesium carbonate (1.3 g,4.0 mmol) and N-methylpiperazine (308.4 mg,3.08 mmol) were added under nitrogen protection, and the reaction mixture was heated to 90℃for 12 hours. After the reaction was completed, the reaction solution was poured into water (20.0 mL), then extracted with ethyl acetate (30.0 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Crude product is subjected to silica gelPurification by chromatography (dichloromethane: methanol=10:1) afforded a brown oil (490.0 mg). LC-MS: [ M+1 ]] + :288.1。
Synthesis of 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) aniline (4):
the compound 1- (4- (difluoromethoxy) -3-nitrophenyl) -4-methylpiperazine (100.0 mg,0.35 mmol) was dissolved in ethyl acetate (2.0 mL), palladium on carbon (20%) (30.0 mg) was added under nitrogen and replaced with hydrogen 3 times. The reaction mixture was heated to 40℃and reacted for 16 hours. After the reaction was completed, the reaction solution was filtered through celite, and the cake was washed twice with a small amount of ethyl acetate, and concentrated to give a yellow solid (65.0 mg). LC-MS: [ M+1 ] ] + :258.2。
Synthesis of 6- (2- ((2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-121):
the compound 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) aniline (50.0 mg,0.20 mmol), 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-7) (56.7 mg,0.19 mmol), palladium acetate (5.8 mg,0.026 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (24.2 mg,0.04 mmol) were dissolved in anhydrous 1, 4-dioxane (3 mL), cesium carbonate (126.6 mg,0.39 mmol) was added under nitrogen protection, and the reaction mixture was reacted at 70℃for 5 hours. After the completion of the reaction, the reaction solution was poured into an aqueous solution (20 mL), followed by extraction with methylene chloride (20 mL. Times.3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to dryness, and purified by preparative separation to give a yellow solid (21.5 mg). 1 H NMR(400MHz,DMSO)δ8.82(s,1H),8.65(d,J＝3.6Hz,2H),8.23–8.19(m,2H),7.82(d,J＝8.0Hz,1H),7.67(s,1H),7.09(d,J＝8.8Hz,1H),6.96(t,J＝74.8Hz,1H),6.70(d,J＝9.2Hz,1H),3.12(s,4H),2.43(s,4H),2.21(s,3H),1.49(s,6H)；LC-MS:[M+1] + :513.3。
EXAMPLE 24 Synthesis of Compounds DP-01-140
Synthesis of tert-butyl 5- (2- ((5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) amino) -5-fluoropyrimidin-4-yl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylate (2):
to a solution of 5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-amine (common int-13) (200.0 mg,0.77 mmol) and tert-butyl 5- (2-chloro-5-fluoropyrimidin-4-yl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylate (303.4 mg,0.77 mmol) in toluene (10 mL) was added successively 1,1 '-binaphthyl-2, 2' -diphenylphosphine (96.39 mg,0.16 mmol), tris (dibenzylideneacetone) dipalladium (141.8 mg,0.16 mmol) and cesium carbonate (756.6 mg,2.32 mmol), and the reaction was stirred under nitrogen at 80℃for 2 hours. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane: methanol=0-10%) to give a white solid (315.0 mg). LC-MS: [ M+H ] ] + :614.2
Synthesis of 6- (2- ((5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (DP-01-140):
5- (2- ((5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) amino) -5-fluoropyrimidin-4-yl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (315.0 mg,0.51 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (2 mL) was added to the reaction solution and stirred for 1 hour at room temperature. The crude product obtained by concentrating the reaction solution was purified by preparation to obtain a white solid (140.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.86(s,2H),8.80(s,1H),8.28–8.25(m,2H),7.95(s,2H),7.85(d,J＝8.0Hz,1H),7.03(t,J＝73.6Hz,1H),3.45(s,4H),2.39(s,4H),2.20(s,3H),1.51(s,6H)；LC-MS:[M+H] + :514.2。
EXAMPLE 25 Synthesis of Compounds PR-01-105
Synthesis of 5-bromo-3-iodo-2-methylbenzoic acid methyl ester (2):
2-methyl-3-amino-5-bromobenzoic acid methyl ester (5.0 g,20.48 mmol) was dissolvedSolution sodium nitrite (1.6 g,23.19 mmol) was dissolved in water (20 mL) and added dropwise to the reaction system at-5℃in a 6N aqueous hydrochloric acid (60 mL,360.00 mmol), sodium iodide (3.4 g,22.68 mmol) was dissolved in water (20 mL) and added dropwise thereto slowly after 30 minutes of reaction, and the reaction solution was warmed to room temperature and reacted at 90℃for 1 hour. The reaction was cooled, extracted with ethyl acetate (15 ml×3), and the organic phase was washed with saturated aqueous sodium chloride (20 mL), dried and spun-dried, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1) to give a brown solid (4.8 g). 1 H NMR(400MHz,DMSO)δ8.25(d,J＝2.0Hz,1H),7.86(d,J＝2.0Hz,1H),3.84(s,3H),2.49(s,3H)。
Synthesis of methyl 5-bromo-2- (bromomethyl) -3-iodobenzoate (3):
methyl 5-bromo-3-iodo-2-methylbenzoate (4.8 g,13.38 mmol) was dissolved in carbon tetrachloride (70 mL), and then N-bromosuccinimide (2.4 g,13.48 mmol) and dibenzoyl peroxide (320.0 mg,1.32 mmol) were added sequentially, and the reaction was allowed to warm to 80℃for 18 hours, cooled to react, and the solvent was dried by spin-drying, and the crude product was subjected to column chromatography (ethyl acetate: petroleum ether=1:10) to give a yellow solid (5.5 g). 1 H NMR(400MHz,DMSO)δ8.38(d,J＝2.1Hz,1H),8.00(d,J＝2.1Hz,1H),4.97(s,2H),3.88(s,3H)。
Synthesis of 6-bromo-4-iodo-isoindol-1-one (4):
methyl 5-bromo-2- (bromomethyl) -3-iodobenzoate (5.5 g,12.68 mmol) was dissolved in 7M methanolic ammonia (60 mL) and the reaction stirred at room temperature for 2 hours. The reaction mixture was dried by spinning, diluted with saturated aqueous sodium hydrogencarbonate (20 mL), extracted with methylene chloride (20 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered and dried by spinning to give a yellow solid (4.2 g). LC-MS: [ M+H ]] + :338.1,340.1。
Synthesis of 6-bromo-4-iodo-1-oxoisoindole-2-carboxylic acid tert-butyl ester (5):
6-bromo-4-iodoisoindol-1-one (4.2 g,12.43 mmol), 4-dimethylaminopyridine (460.0 mg,3.77 mmol) and triethylamine (5.0 g,49.41 mmol) were dissolved in dichloromethane (100 mL), and di-tert-butyl dicarbonate (8.1 g,37.11 mmol) was slowly added dropwise to the reaction system and reacted at room temperature for 18 hours. The reaction system was diluted with dichloromethane (50 mL), washed with water (100 mL), and the organic phase was separated and dried Drying, filtering and spin drying. The crude product was subjected to column chromatography (petroleum ether: ethyl acetate=5:1) to give a white solid (2.7 g): LC-MS: [ M ] t Bu+H] + :382.1,384.1。
Synthesis of 6-bromo-4-iodo-3-methyl-1-oxoisoindole-2-carboxylic acid tert-butyl ester (6):
6-bromo-4-iodo-1-oxoisoindole-2-carboxylic acid tert-butyl ester (1.2 g,2.74 mmol) was dissolved in tetrahydrofuran (60 mL), 1M lithium bis trimethylsilylamide (3.3 mL,3.30 mmol) was added dropwise at-70℃and stirred at-70℃for half an hour. Methyl iodide (580.0 mg,4.09 mmol) was added dropwise to the reaction system, the reaction was stirred for 0.5 hours after warming to room temperature, the reaction was quenched with saturated aqueous ammonium chloride (20 mL), extracted with ethyl acetate (20 mL. Times.3), and the organic phase was washed with saturated aqueous sodium chloride (20 mL), dried and spun-dried to give a white solid (1.1 g). LC-MS: [ M ] t Bu+H] + :396.0,398.0。
Synthesis of 5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (7):
6-bromo-4-iodo-3-methyl-1-oxoisoindole-2-carboxylic acid tert-butyl ester (1.1 g,2.43 mmol) was dissolved in tetrahydrofuran (60 mL), 1M lithium bis trimethylsilylamide (2.9 mL,2.90 mmol) was added dropwise with stirring at-70℃and stirring at-70℃for half an hour. Methyl iodide (1.0 g,7.05 mmol) was added dropwise to the reaction system, the reaction was warmed to room temperature and stirred for 0.5 hours, the reaction was quenched with saturated aqueous ammonium chloride (50 mL), extracted with ethyl acetate (50 mL. Times.3), the organic phase was washed with saturated aqueous sodium chloride (100 mL), dried, and spun-dried, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1) to give a white solid (180.0 mg), LC-MS: [ M-tBu+H ] + :410.0,412.0。
(E) -synthesis of 5-bromo-7- (2-ethoxyvinyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (8):
tert-butyl 5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindole-2-carboxylate (250.0 mg,0.54 mmol), (E) -2- (2-ethoxyvinyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan (130.0 mg,0.66 mmol), potassium phosphate (250.0 mg,1.18 mmol) and 1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (40.0 mg,0.06 mmol) was dissolved in dioxane (5 mL) and water (0.5 mL), under nitrogen blanket, 10The reaction was carried out at 0℃for 3 hours. The reaction system was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=10:1) to give a white solid (150.0 mg). 1 H NMR(400MHz,CDCl 3 )δ7.81(s,1H),7.62(s,1H),6.88(d,J＝12.0Hz,1H),6.04(d,J＝12.4Hz,1H),3.97(q,J＝7.2Hz,2H),1.80(s,6H),1.61(s,9H),1.38(t,J＝7.2Hz,3H).
Synthesis of 2- (6-bromo-3, 3-dimethyl-1-oxoisoindol-4-yl) acetaldehyde (9):
(E) -5-bromo-7- (2-ethoxyvinyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (150.0 mg,0.37 mmol) was dissolved in tetrahydrofuran (3 mL), concentrated hydrochloric acid (1 mL,0.24 mmol) was added dropwise at room temperature and stirred for 3 hours. To the reaction system was added dropwise saturated aqueous sodium bicarbonate until ph=9. Extraction with ethyl acetate (5 mL. Times.3), washing the organic phase with saturated brine (10 mL), drying, and spin-drying gave a white crude product (80.0 mg). LC-MS: [ M+H ] ] + :282.2,284.2。
Synthesis of 6-bromo-4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (10):
the compound 2- (6-bromo-3, 3-dimethyl-1-oxoisoindol-4-yl) acetaldehyde (80.0 mg,0.28 mmol) was dissolved in ethanol (4 mL), sodium borohydride (85.0 mg,2.25 mmol) was added at room temperature and reacted for 1 hour. The reaction was quenched with ice water (8 mL), extracted with ethyl acetate (8 mL. Times.3), the organic phase was washed with saturated aqueous sodium chloride (8 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a white crude product (80.0 mg). LC-MS: [ M+H ]] + :284.1,286.1。
Synthesis of 4- (2-hydroxyethyl) -3, 3-dimethyl-6-boronic acid pinacol ester-isoindol-1-one (11):
the compound 6-bromo-4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (80.0 mg,0.28 mmol), pinacol biborate (85.0 mg,0.34 mmol), potassium acetate (70.0 mg,0.71 mmol) and 1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (20.0 mg,0.03 mmol) was suspended in anhydrous dioxane (4 mL), reacted at 100℃for 18 hours under nitrogen protection, water (10 mL) was added to the system, followed by extraction with ethyl acetate (10 mL. Times.3), and the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a brown crude product (80.0 mg). LC-MS: [ M+H ]] + :332.0。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (12):
The compound 4- (2-hydroxyethyl) -3, 3-dimethyl-6-boronic acid pinacol ester-isoindol-1-one (80.0 mg,0.24 mmol), 2, 4-dichloro-5-fluoropyrimidine (50.0 mg,0.30 mmol), potassium phosphate (125.0 mg,0.59 mmol) and 1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (17.0 mg,0.02 mmol) was suspended in anhydrous dioxane (4 mL) and water (0.8 mL) and reacted at 100℃for 2 hours under nitrogen. The reaction was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol=10:1) to give a brown solid (60.0 mg). LC-MS: [ M+H ]] + :336.3。
Synthesis of 6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (PR-01-105):
6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (45.0 mg,0.13 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (37.0 mg,0.13 mmol), cesium carbonate (120.0 mg,0.37 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (20.0 mg,0.03 mmol) and palladium acetate (4.5 mg,0.02 mmol) were dissolved in dioxane (2 mL), the reaction system was dried by spin-drying at 100℃under nitrogen protection for 18 hours, and the crude product was prepared as a white solid (15.5 mg) by basicity. 1 H NMR(400MHz,DMSO-d6)δ9.10(s,1H),8.79(s,1H),8.62(d,J＝3.2Hz,1H),8.04(s,2H),7.46(s,1H),7.20(d,J＝8.8Hz,1H),6.77(d,J＝9.2Hz,1H),4.90(s,1H),3.68(s,2H),3.15(s,4H),2.99(t,J＝6.8Hz,2H),2.44(s,4H),2.21(s,3H),2.07(s,1H),1.56(s,6H)；LC-MS:[M+H] + :575.5。
EXAMPLE 26 Synthesis of Compound PR-01-119
Synthesis of 5-bromo-4-iodo-2- (trifluoromethoxy) aniline (2):
5-bromo-2- (trifluoromethoxy) aniline (1.0 g,3.91 mmol) and N-iodosuccinimide (970.0 mg,4.30 mmol) were dissolved in N, N-dimethylformamideAfter reacting for 8 hours at room temperature, ethyl acetate (60 mL) was added to the amine (20 mL) for dilution. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. Purification by column chromatography on silica gel (petroleum ether/ethyl acetate=5/1) gave a brown liquid (810.0 mg). LC-MS: [ M+H ]] + :381.9,383.9。
Synthesis of 4-amino-2-bromo-5- (trifluoromethoxy) benzonitrile (3):
the compound 5-bromo-4-iodo-2- (trifluoromethoxy) aniline (420.0 mg,1.10 mmol), zinc cyanide (64.6 mg,0.55 mmol), tetrakis (triphenylphosphine) palladium (127.1 mg,0.11 mmol) was dissolved in N, N-dimethylformamide (4 mL) and heated to 90℃under nitrogen for 16 hours. Ethyl acetate (20 mL) was added to the reaction mixture to dilute it, and the mixture was filtered. The filtrate was washed three times with water and saturated brine in this order, the organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/3) to give a yellow solid product (290.0 mg). LC-MS [ M-H ]] - :279.0,281.0。
Synthesis of 2-bromo-4- ((4- (1, 1-dimethyl-3-oxoisoindol-5-yl) -5-fluoropyrimidin-2-yl) amino) -5- (trifluoromethoxy) benzonitrile (4):
4-amino-2-bromo-5- (trifluoromethoxy) benzonitrile (200.0 mg,0.71 mmol), 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-7) (249.1 mg,0.85 mmol) and cesium carbonate (695.6 mg,2.14 mmol) were dissolved in N, N-dimethylformamide (5 mL), and the reaction mixture was stirred at 80℃for 6 hours. After cooling to room temperature, ethyl acetate (40 mL) was added to dilute the mixture, and the organic phase was washed with saturated aqueous sodium chloride (30 ml×3), dried over anhydrous sodium sulfate, concentrated, and chromatographed on silica gel (PE/ea=1/1) to give a white solid (210.0 mg). LCMS [ M+H ]] + :536.2,538.2。
Synthesis of 4- ((4- (1, 1-dimethyl-3-oxoisoindol-5-yl) -5-fluoropyrimidin-2-yl) amino) -2- (4-methylpiperazin-1-yl) -5-5- (trifluoromethoxy) benzonitrile (PR-01-119):
2-bromo-4- ((4- (1, 1-dimethyl-3-oxoisoindol-5-yl) -5-fluoropyrimidin-2-yl) amino) -5- (trifluoromethoxy) benzonitrile (30.0 mg,0.06 mmol), 1-methylpiperazine (11.2 mg,0.11 mmol), tris (dibenzylideneacetone) dipalladium (9.2 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (3.9 mg,0.01 mmol) was suspended in tetrahydrofuran (1 mL). 1M lithium bis (trimethylsilylamide) (0.2 mL,0.20 mmol) was slowly added at room temperature under argon. After the reaction was heated to 70 ℃ and stirred for 2 hours LCMS detected completion of the reaction. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, and concentrated. Purification by basicity gave a white solid (6.0 mg). 1 H NMR(400MHz,Methanol-d 4 )δ8.63(d,J＝3.2Hz,1H),8.59(s,1H),8.46(s,1H),8.39(d,J＝8.2Hz,1H),7.801(d,J＝8.0Hz,1H),7.618(s,1H),3.31(s,4H),2.67(s,4H),2.37(s,3H),1.60(s,6H)；LCMS:[M+H] + :556.1。
EXAMPLE 27 Synthesis of Compound PR-01-116A
Synthesis of 6- (5-fluoro-2- ((5- (1-methyl-4-oxo-1, 4-azaphosphino-4-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-116A):
the compound 1-methyl-1, 4-azaphosphine-4-oxide (40.0 mg,0.30 mmol), 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8) (153.8 mg,0.30 mmol), tris (dibenzylideneacetone) dipalladium (27.5 mg,0.03 mmol), 4, 5-diphenylphosphine-9, 9-dimethylxanthene (17.4 mg,0.03 mmol), potassium phosphate (127.2 mg,0.6 mmol) were dissolved in N, N-dimethylformamide (2 mL) and reacted at 100 ℃ for 4 hours under nitrogen protection. The reaction solution was cooled and filtered, the filtrate was diluted with water (30 mL), extracted with ethyl acetate (30 mL. Times.3), and concentrated to a black-like oil, which was purified by preparative HPLC to give a white solid (60 mg). 1 H NMR(400MHz,MeOD)δ8.92(d,J＝12.8Hz,1H),8.56(s,1H),8.43–8.40(m,2H),7.78(d,J＝7.4Hz,1H),7.59–7.57(m,2H),3.00–2.88(m,2H),2.88–2.79(m,2H),2.37(s,3H),2.33–2.30(m,2H),2.13(t,J＝16.4Hz,2H),1.59(s,6H)；LC-MS:[M+H] + :564.3。
EXAMPLE 28 Synthesis of Compounds PR-01-109
Synthesis of 6- (5-fluoro-2- ((5- (4-isopropylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-109):
the compound 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8) (50.0 mg,0.10 mmol), 1-isopropylpiperazine (19.2 mg,0.15 mmol), tris (dibenzylideneacetone) palladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M lithium bistrimethylsilylamino (0.49 mL,0.49 mmol) were dissolved in tetrahydrofuran (0.5 mL) and reacted at 70 ℃ for 6 hours under nitrogen protection. The reaction was cooled, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (2 mL). The organic phase is dried, spun-dried, and then subjected to alkaline (H 2 O/MeCN(0.1％ NH 3 ·H 2 O)) was prepared, isolated and purified to give a white solid (14.1 mg). 1 H NMR(400MHz,MeOD)δ8.51(d,J＝3.3Hz,1H),8.43(s,1H),8.35(d,J＝7.9Hz,1H),8.20(d,J＝2.6Hz,1H),7.74(d,J＝8.0Hz,1H),7.19(d,J＝8.9Hz,1H),6.68(m,1H),3.26–3.20(m,4H),2.71–2.70(m,5H),1.59(s,6H),1.12(d,J＝6.5Hz,6H)；LCMS:[M+H] + :559.4。
EXAMPLE 29 Synthesis of Compounds PR-01-110
Synthesis of 6- (2- ((5- (4-cyclopropylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-110):
the compound 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8) (50 mg,0.10 mmol), N-cyclopropylpiperazine (18.9 mg,0.15 mmol), tris (dibenzylideneacetone) palladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M lithium bis trimethylsilylamide (0.49 mL, 0.4)9 mmol) was dissolved in tetrahydrofuran (0.5 mL) and reacted at 70℃for 6 hours under nitrogen. The reaction was cooled, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (2 mL). The organic phase was dried, spun-dried, and purified by preparative separation to give a white solid (13.5 mg). 1 H NMR(400MHz,DMSO)δ9.08(s,1H),8.81(s,1H),8.64(d,J＝3.2Hz,1H),8.19–8.18(m,2H),7.80(d,J＝8.0Hz,1H),7.56(d,J＝2.4Hz,1H),7.20(d,J＝8.8Hz,1H),6.73(dd,J＝9.2,2.8Hz,1H),3.10(d,J＝4.8Hz,4H),2.65(d,J＝4.8Hz,4H),1.65(s,1H),1.49(s,6H),0.43–0.42(m,2H),0.34(d,J＝2.8Hz,2H)；LC-MS:[M+1] + :557.2。
EXAMPLE 30 Synthesis of Compounds PR-01-111
Synthesis of 1- (2- ((tert-butyldimethylsilyloxy) ethyl) piperazine (2):
to a solution of N-hydroxyethyl piperazine (1.9 mL,15.37 mmol) and 1H-imidazole (1.6 g,23.50 mmol) in tetrahydrofuran (15 mL) at 0deg.C under nitrogen, tert-butyldimethylchlorosilane (3.5 g,23.22 mmol) was added dropwise, stirred overnight at room temperature, and the reaction mixture was concentrated in vacuo to remove most of the solvent to give the crude product. The crude product was chromatographed (dichloromethane: methanol=0-70%) to give a white solid (900.0 mg). LC-MS [ M+H-t-Bu ] ] + :189.3。
Synthesis of 5- (4- (2- ((tert-butyldimethylsilyloxy) ethyl) piperazin-1-yl) -2- (trifluoromethoxy) aniline (3):
to a solution of 5-bromo-2- (trifluoromethoxy) aniline (200.0 mg,0.78 mmol), tris (dibenzylideneacetone) dipalladium (36.6 mg,0.04 mmol) and 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (31.5 mg,0.08 mmol) in tetrahydrofuran (2 mL) at 25 ℃ under nitrogen was added 1M lithium bis (trimethylsilylamide) (2.3 mL,2.34 mmol) and 1- (2- ((tert-butyldimethylsilyloxy) ethyl) piperazine (381.9 mg,1.56 mmol), and the resulting mixture was stirred at 80 ℃ for 6 hours, the reaction mixture was cooled to room temperature and concentrated in vacuo to remove most of the solvent. The residue was poured into water (5 mL) and extracted with ethyl acetate (10 mL)X 3) extraction. The organic layer was dried over anhydrous sodium sulfate, and the solid was subjected to column chromatography (dichloromethane: methanol=10:1, rf=0.5) to give a white solid (50 mg). LC-MS [ M+H-t-Bu ]] + :364.5。
Synthesis of 6- (2- ((5- (4- (2- (tert-butyldimethylsilyl) oxy) ethyl) piperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (4):
to a solution of 5- (4- (2- ((tert-butyldimethylsilyloxy) ethyl) piperazin-1-yl) -2- (trifluoromethoxy) aniline (30.0 mg,0.07 mmol) and 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-7) (20.9 mg,0.07 mmol) in dioxane (5 mL) was added cesium carbonate (45.6 mg,0.14 mmol) followed by N 2 Palladium acetate (2.2 mg,0.01 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (6.2 mg,0.01 mmol) were added with stirring. The mixture was refluxed for 16 hours, the reaction mixture was cooled to room temperature and concentrated in vacuo to remove most of the solvent. The residue was poured into water (3 mL) and extracted with ethyl acetate (5 ml×3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The solid was subjected to column chromatography (dichloromethane: methanol=10:1, rf=0.3) to give a white solid (20.0 mg). LC-MS [ M+H-t-Bu ]] + :619.3。
Synthesis of 6- (5-fluoro-2- ((5- (4- (2-hydroxyethyl) piperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-111):
6- (2- ((5- (4- (2- (tert-butyldimethylsilyl) oxy) ethyl) piperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (50.0 mg,0.07 mmol) was added to tetrahydrofuran (1 mL) at room temperature, under N 2 1M tetrabutylammonium fluoride (1 mL,1 mmol) was added. The resulting mixture was stirred at 25 ℃ for 5 hours, the reaction solution was concentrated in vacuo to remove most of the solvent, and the solid was made basic to give a white solid (11.2 mg). 1 H NMR(400MHz,Methanol-d 4 )δ8.52(d,J＝3.6Hz,1H),8.44(d,J＝1.2Hz,1H),8.40–8.35(m,1H),8.17(d,J＝2.8Hz,1H),7.74(dd,J＝8.0,0.4Hz,1H),7.19(dd,J＝9.2,1.6Hz,1H),6.69(dd,J＝9.2,2.8Hz,1H),3.72(t,J＝6.0Hz,2H),3.28–3.21(m,4H),2.69(t,J＝5.2Hz,4H),2.59(t,J＝5.6Hz,2H),1.59(s,6H).；LC-MS:[M+H] + :561.1。
EXAMPLE 31 Synthesis of Compounds PR-01-112
Synthesis of 6- (5-fluoro-2- ((5- (4- (2-fluoroethyl) piperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-112):
The compound 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8) (50.0 mg,0.10 mmol), 1- (2-fluoroethyl) -piperazine (20.0 mg,0.15 mmol), tris (dibenzylideneacetone) palladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M bistrimethylsilylamino lithium (0.5 mL,0.50 mmol) were dissolved in tetrahydrofuran (0.5 mL) and reacted at 70 ℃ for 6 hours under nitrogen protection. The reaction was cooled, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (2 mL). The organic phase was dried and spun-dried and the crude product prepared on a silica gel plate (petroleum ether: ethyl acetate: dichloromethane: methanol=10:10:10:1) to give a white solid (5.0 mg). 1 H NMR(400MHz,MeOD)δ8.52(s,1H),8.44(s,1H),8.37(d,J＝7.6Hz,1H),8.19(s,1H),7.75(d,J＝8.0Hz,1H),7.19(d,J＝8.8Hz,1H),6.69(d,J＝9.2Hz,1H),4.67(s,1H),4.55(s,1H),3.25(s,4H),2.80(s,1H),2.71(s,5H),1.59(s,6H)；LC-MS:[M+H] + :563.4。
EXAMPLE 32 Synthesis of Compounds PR-01-114
Synthesis of 6- (5-fluoro-2- ((5- (3-methyl-3, 6-diazabicyclo [3.1.1] heptan-6-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-114):
the compound 6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common)int-8) (50.0 mg,0.10 mmol), 3-methyl-3, 7-diazabicyclo [3.1.1]]Heptane (16.8 mg,0.15 mmol), tris (dibenzylideneacetone) palladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M lithium bis trimethylsilylamide (0.49 mL,0.49 mmol) were dissolved in tetrahydrofuran (0.5 mL) and reacted at 70℃for 6 hours under nitrogen. The reaction was cooled, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (2 mL). The organic phase was dried, spun-dried, and the crude product was made basic in reverse phase to give a white solid (8.1 mg). 1 H NMR(400MHz,MeOD)δ8.53(s,1H),8.40(d,J＝8.4Hz,2H),8.32(d,J＝8.0Hz,1H),7.76(s,2H),7.24(d,J＝8.0Hz,1H),6.29(d,J＝8.4Hz,1H),4.39(d,J＝5.2Hz,2H),3.46(d,J＝13.2Hz,2H),3.27(s,1H),2.79–2.77(m,1H),2.47(s,3H),2.04(d,J＝8.8Hz,1H),1.59(s,6H)；LC-MS:[M+H] + :543.5。
EXAMPLE 33 Synthesis of Compounds PR-01-123, PR-01-123A, PR-01-123B
Synthesis of 6- (2- ((5- (3, 4-dimethylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (PR-01-123):
6- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) -3, 3-dimethylisoindol-1-one (common int-8) (50.0 mg,0.10 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (3.9 mg,0.01 mmol), tris (dibenzylideneacetone) dipalladium (9.2 mg,0.01 mmol) were added to tetrahydrofuran (1.5 mL) and replaced three times with nitrogen. 1, 2-dimethylpiperazine (22.8 mg,0.20 mmol) and 1M lithium bis trimethylsilylamide (0.3 mL,0.3 mmol) were then added to the reaction solution. After stirring the reaction at 70 ℃ for 5 hours LCMS detected completion of the reaction. The reaction mixture was poured into water (5 mL), extracted with ethyl acetate (5 mL. Times.3), and the organic phases were combined and concentrated to dryness. Basic prep HPLC purification gave a pale yellow solid (20.0 mg). 1 H NMR(400MHz,Methanol-d 4 )δ8.51(d,J＝3.2Hz,1H),8.44(s,1H),8.37(d,J＝8.0Hz,1H),8.12–8.10(m,1H),7.75(d,J＝8.0Hz,1H),7.19(d,J＝8.8Hz,1H),6.70–6.67(m,1H),3.58(d,J＝12.0Hz,1H),3.50(d,J＝12.0Hz,1H),2.93–2.86(m,2H),2.55–2.41(m,2H),2.33(s,4H),1.58(s,6H),1.05(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :545.2。
The racemate PR-01-123 was purified by SFC separation (column: chiralPakC-IG, 250X 21.2mm I.D.,5 μm; mobile phase: A phase: carbon dioxide, B phase: [ MEOH+0.1% MEA ]; gradient: B:45% -45%,8min;120 min) to give white solids PR-01-123A and PR-01-123B.
PR-01-123A：
Chiral analysis conditions: chromatographic column: CHIRALPAK IB 100 x 4.6mm 5 μm; mobile phase: phase A: carbon dioxide; and B phase: meOH (0.05% DEA v/v); gradient: b:40% -40%; retention time: 2.987min; ee% = 100%.
1 H NMR(400MHz,Methanol-d 4 )δ8.51(d,J＝3.2Hz,1H),8.44(s,1H),8.37(d,J＝8.0Hz,1H),8.11(d,J＝3.2Hz,1H),7.76(d,J＝8.0Hz,1H),7.19(d,J＝9.2Hz,1H),6.68(d,J＝8.8Hz,1H),3.59(d,J＝11.6Hz,1H),3.50(d,J＝12.0Hz,1H),2.91–2.85(m,2H),2.57–2.43(m,2H),2.33–2.32(m,4H),1.58(s,6H),1.04(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :545.4。
PR-01-123B：
Chiral analysis conditions: chromatographic column: CHIRALPAK IB 100 x 4.6mm 5 μm; mobile phase: phase A: carbon dioxide; and B phase: meOH (0.05% DEAVV/v); gradient: b:40% -40%; retention time: 4.407min; ee% = 100%.
1 H NMR(400MHz,Methanol-d 4 )δ8.51(d,J＝2.8Hz,1H),8.44(s,1H),8.37(d,J＝8.0Hz,1H),8.11(s,1H),7.76(d,J＝8.4Hz,1H),7.19(d,J＝9.2Hz,1H),6.68(d,J＝9.2Hz,1H),3.59(d,J＝12.0Hz,1H),3.49(d,J＝12.4Hz,1H),2.91–2.85(m,2H),2.53–2.40(m,2H),2.325(s,4H),1.59(s,6H),1.04(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :545.4。
EXAMPLE 34 Synthesis of Compounds PR-01-080
Synthesis of 5' - (5-chloro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) -1, 2-dihydropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindol ] -3' -one (PR-01-080):
spiro [ cyclopropane-1, 1 '-isoindole 5' -boronic acid pinacol ester]-3 '-Ketone (common int-9) (100.0 mg,0.35 mmol), potassium carbonate (144.9 mg,1.05 mmol), 4, 5-dichloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) -1, 2-dihydropyrimidin-2-amine (common int-5) (176.8 mg,0.42 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (29.1 mg,0.04 mmol) was dissolved in 1, 4-dioxane (20 mL) and water (3 mL). The reaction was stirred at 100 ℃ for 16 hours under nitrogen, LCMS showed complete reaction. The reaction solution was poured into water (15 mL), followed by extraction with ethyl acetate (3×15 mL), and the organic phase was washed with saturated brine, followed by separation of the organic phase. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (ethyl acetate: petroleum ether=0-100%, rf=0.6) to give the crude product. The crude product was purified by prep. to give a white solid (30.0 mg). 1 H NMR(400MHz,CDCl 3 )δ8.51(s,1H),8.43(s,1H),8.32(d,J＝2.8Hz,1H),8.10(dd,J＝8.0,1.6Hz,1H),7.49(s,1H),7.21–7.11(m,2H),6.94(s,1H),6.54(dd,J＝9.2,2.8Hz,1H),3.24–3.18(m,4H),2.57–2.50(m,4H),2.34(s,3H),1.64(s,4H)；LCMS:[M+H] + :545.5。
Example 35 Synthesis of Compounds PR-01-106:
synthesis of 5' - (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindol ] -3' -one (PR-01-106):
the compound 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindole]-3' -Ketone (common int-10) (150.0 mg,0.52 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (155.0 mg,0.56 mmol), cesium carbonate (506.0 mg,1.55 mmol), tris (dibenzylideneacetone) dipalladium (47.4 mg,0.05 mmol), 1'-binaphthyl-2, 2' -bisdiphenylphosphine (64.5 mg,0.10 mmol) was suspended in toluene (10 mL) and the reaction mixture was reacted at 120℃for 6 hours under nitrogen protection. The reaction solution was cooled to room temperature, poured into water (20 mL), extracted with ethyl acetate (15 ml×3), and the combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated, followed by silica gel column chromatography (dichloromethane: methanol=10:1, rf=0.6) to give a crude product. The crude product was purified by prep. to give a white solid (43.5 mg). 1 H NMR(400MHz,DMSO-d6)δ9.08(s,1H),8.87(s,1H),8.63(d,J＝3.6Hz,1H),8.29(s,1H),8.19(d,J＝8.4Hz,1H),7.51(d,J＝2.8Hz,1H),7.47(d,J＝8.4Hz,1H),7.22–7.19(m,1H),6.75(dd,J＝9.2,2.8Hz,1H),3.16(t,J＝4.4Hz,4H),2.45–2.43(m,4H),2.21(s,3H),1.53(s,4H)；LC-MS:[M+H] + :529.2。
EXAMPLE 36 Synthesis of Compound PR-01-117
Synthesis of 2-chloro-4-iodo-5- (trifluoromethoxy) pyridine (2):
the compound 2-chloro-5- (trifluoromethoxy) pyridine (1.0 g,5.06 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), and after cooling to-78deg.C, lithium diisopropylamide (3.1 mL,6.08mmol,2 mol/L) was slowly added dropwise, and after maintaining the temperature for 1 hour stirring, a solution of iodine (1.41 g,5.56 mmol) in tetrahydrofuran (5 mL) was added dropwise. The reaction mixture was gradually warmed to room temperature and stirred for 2 hours. LCMS monitored reaction was complete. The reaction solution was poured into an aqueous ammonium chloride solution (20 mL), and extracted with ethyl acetate (20 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Purification by silica gel column chromatography (petroleum ether: ethyl acetate=5:1, rf=0.4) afforded a yellow solid (980.0 mg). 1 H NMR(400MHz,CDCl 3 )δ8.25(s,1H),7.88(s,1H)。
Synthesis of N- (2-chloro-5- (trifluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethanimine (3):
the compound 2-chloro-4-iodo-5- (trifluoromethoxy) pyridine (840.0 mg,2.60 mmol), benzophenone imine (470.0 mg,2.60 mmol), tris (dibenzylideneacetone) dipalladium (238.0 mg, 0.2)6 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (323.0 mg,0.52 mmol), cesium carbonate (2.5 g,7.67 mmol) were dissolved in anhydrous toluene (10 mL). The reaction was carried out at 60℃for 5 hours under nitrogen. LCMS monitored complete reaction of starting material. The reaction solution was directly concentrated and purified by silica gel column chromatography (ethyl acetate: petroleum ether=5:1, rf=0.5) to give a yellow oil (750.0 mg). LC-MS: [ M+1 ]] + :377.2,379.2.
Synthesis of N- (2- (4-methylpiperazin-1-yl) -5- (trifluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethanimine (4):
the compound N- (2-chloro-5- (trifluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethane imine (800.0 mg,2.12 mmol), 1-methylpiperazine (213.0 mg,2.13 mmol), tris (dibenzylideneacetone) dipalladium (195.0 mg,0.21 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (172.0 mg,0.42 mmol), sodium t-butoxide (612.0 mg,6.37 mmol) was dissolved in anhydrous toluene (10 mL), replaced with nitrogen three times, and the reaction was heated to 110℃and stirred for 16 hours. LCMS monitored reaction was complete. The reaction solution was directly concentrated and purified by silica gel column chromatography (dichloromethane: methanol=10:1, rf=0.5) to give a yellow solid (680.0 mg). LC-MS: [ M+1 ] ] + :441.5。
Synthesis of 2- (4-methylpiperazin-1-yl) -5- (trifluoromethoxy) pyridin-4-amine (5):
the compound N- (2- (4-methylpiperazin-1-yl) -5- (trifluoromethoxy) pyridin-4-yl) -1, 1-diphenylmethanimine (680.0 mg,1.54 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (2 mL) was added dropwise under nitrogen. The reaction was stirred at room temperature for 4 hours and LCMS monitored to complete the reaction. The reaction solution was directly concentrated and then purified by column chromatography (dichloromethane: methanol=15:1) to give a yellow oil (400.0 mg). LC-MS: [ M+1 ]] + :277.4。
Synthesis of 5' - (5-fluoro-2- ((2- (4-methylpiperazin-1-yl) -5- (trifluoromethoxy) pyridin-4-yl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (PR-01-117):
the compound 2- (4-methylpiperazin-1-yl) -5- (trifluoromethoxy) pyridin-4-amine (120.0 mg,0.43 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindole]3' -one (common int-10) (126.0 mg,0.43 mmol), tris (dibenzylideneacetone) dipalladium (40).0mg,0.04 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (54.0 mg,0.087 mmol) and cesium carbonate (425.0 mg,1.30 mmol) were dissolved in anhydrous toluene (5 mL), and the reaction mixture was heated to 100℃for 6 hours after nitrogen substitution. The reaction solution was directly concentrated and then subjected to column chromatography (dichloromethane: methanol=5:1) to give a crude product. The crude product was purified by prep HPLC to give an off-white solid (22.0 mg). 1 H NMR(400MHz,DMSO-d6)δ9.51(s,1H),8.91(s,1H),8.79(s,1H),8.34(s,1H)，8.25(d,J＝8.0Hz,1H),8.06(s,1H),7.85(s,1H),7.51(d,J＝8.0Hz,1H),3.48(s,4H),2.39(s,4H),2.21(s,3H),1.55(s,4H)；LC-MS:[M+1] + :530.4。
EXAMPLE 37 Synthesis of Compounds DP-01-135
Synthesis of 4-bromo-1- (difluoromethoxy) -2-nitrobenzene (2):
the compound 4-bromo-2-nitrophenol (1.0 g,4.59 mmol), difluorochloroacetic acid (0.72 g,5.52 mmol), cesium carbonate (4.5 g,13.81 mmol) was suspended in DMF (25.0 mL) and stirred at 80℃for 3h. The reaction mixture was diluted with water (80 mL), and extracted with ethyl acetate (80 mL. Times.3). The combined organic phases were washed with saturated aqueous sodium chloride (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Purification by silica gel column chromatography (petroleum ether: ethyl acetate=0% -5%) gave a white solid (0.6 g). LC-MS: [ M+H ]] + :268.1,270.1。
Synthesis of 1- (4- (difluoromethoxy) -3-nitrophenyl) -4-methylpiperazine (3):
the compound 4-bromo-1- (difluoromethoxy) -2-nitrobenzene (0.6 g,2.24 mmol), 1-methylpiperazine (268.8 mg,2.69 mmol), tris [ dibenzylideneacetone]Dipalladium (201.3 mg,0.22 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (127.2 mg,0.22 mmol), cesium carbonate (1.5 g,4.60 mmol) were suspended in toluene (10 mL) and reacted at 90℃for 12h. The reaction solution was purified by silica gel column chromatography (petroleum ether: ethyl acetate=0% -35%) to give a white solid (340.0 mg). LC-MS: [ M+H ]] + :288.1。
Synthesis of 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) aniline (4):
The compound 1- (4- (difluoromethoxy) -3-nitrophenyl) -4-methylpiperazine (290.0 mg,1.01 mmol), pd/C (100 mg,10% w/w) was suspended in ethyl acetate (5 mL) and reacted under a hydrogen atmosphere (1 atm) at 40℃for 2h. The reaction solution was filtered, and the cake was washed with ethyl acetate (50 mL. Times.3). The filtrate was concentrated to give crude product (250.0 mg) as pale yellow solid. LC-MS: [ M+H ]] + :258.1。
Synthesis of 5' - (2- ((2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-135):
the compound 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) aniline (100.0 mg,0.39 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (112.5 mg,0.39 mmol), palladium acetate (9.6 mg,0.04 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (24.8 mg,0.04 mmol), cesium carbonate (253.5 mg,0.78 mmol) were dissolved in 1, 4-dioxane (1 mL), and heated to 90℃under nitrogen to react for 5 hours. The reaction mixture was cooled, diluted with water (30 mL), and extracted with ethyl acetate (30 mL. Times.3). The organic phases are combined and washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a black solid crude product. The crude product was purified by prep HPLC to give a white solid (48.6 mg). 1 H NMR(400MHz,DMSO-d6)δ8.88(s,1H),8.65–8.63(m,2H),8.29(s,1H),8.20(dd,J＝7.2,1.6Hz,1H),7.65(d,J＝2.8Hz,1H),7.47(d,J＝8.0Hz,1H),7.14–6.77(m,2H),6.69(dd,J＝9.2,2.8Hz,1H),3.13(t,J＝4.8Hz,4H),2.46(t,J＝4.8Hz,4H),2.23(s,3H),1.54(s,4H)；LC-MS:[M+H] + :511.2。
EXAMPLE 38 Synthesis of Compounds DP-01-138
Synthesis of (5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-yl) -5-fluoro-4- (spiro [ cyclopropane-1, 1 '-isoindoline ] -5' -yl) pyrimidin-2-amine (DP-01-138):
to 5- (difluoromethoxy) -2- (4-methylpiperazin-1-yl) pyridin-4-amine (common i)nt-13) (120.0 mg,0.46 mmol) and 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a solution of 3' -ketone (common int-10) (127.9 mg,0.44 mmol) in toluene (10 mL) was added sequentially 1,1' -binaphthyl-2, 2' -bisdiphenylphosphine (57.9 mg,0.09 mmol), tris (dibenzylideneacetone) dipalladium (85.1 mg,0.09 mmol), cesium carbonate (302.8 mg,0.93 mmol). The mixture was stirred at 85 ℃ for 16 hours. The reaction mixture was diluted with water (20 mL), and extracted with ethyl acetate (50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was isolated and purified by preparative HPLC to give a yellow solid (13.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.91(s,1H),8.88(s,1H),8.80(d,J＝3.2Hz,1H),8.35(s,1H),8.26(d,J＝8.0Hz,1H),7.95(s,1H),7.93(s,1H),7.52(d,J＝8.0Hz,1H),7.03(t,J＝73.8Hz,1H),3.49–3.41(m,4H),2.43–2.35(m,4H),2.21(s,3H),1.56(s,4H)；LC-MS:[M+H] + :512.2。
EXAMPLE 39 Synthesis of Compounds DP-01-139
Synthesis of 5-bromo-2- (difluoromethoxy) -3-iodopyridine (2):
to N, N-dimethylformamide (300.0 mL) was added 5-bromo-3-iodopyridin-2-ol (30.4 g,101.37 mmol), cesium carbonate (43.0 g,131.97 mmol), sodium difluorochloroacetate (31.3 g,205.3 mmol) under nitrogen. The reaction mixture was heated to 80℃and stirred for 24 hours, after completion of the reaction, quenched with water (100.0 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. Column chromatography (PE: ea=10:1) gave a pale yellow solid (13.4 g). LC-MS: [ M+H ] ] + :349.9,351.9。
Synthesis of N- (5-bromo-2- (difluoromethoxy) pyridin-3-yl) -1, 1-diphenylmethane imine (3):
to a suspension of 5-bromo-2- (difluoromethoxy) -3-iodopyridine (13.4 g,38.19 mmol), benzophenone imine (9.0 g,49.66 mmol) in N, N-dimethylformamide (130.0 mL) was added cuprous bromide (1.1 g,7.66 mmol), (+ -) -1,1' -bis (2-naphthol) (2.2 g,766 mmol) and potassium phosphate (16.3 g,76.8 mmol). The reaction system was stirred at 70℃for 8 hours under nitrogen protection. After the reaction was completed, the reaction mixture was quenched with water (50.0 mL), extracted with ethyl acetate (100 mL. Times.3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. Purification by column chromatography on silica gel (PE: ea=2:1) gave a brown oily liquid (2.3 g). LC-MS: [ M+H ]] + :403.1,405.1。
Synthesis of N- (2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) pyridin-3-yl) -1, 1-diphenylmethanimine (4):
to a 1, 4-dioxane suspension (25.0 mL) of N- (5-bromo-2- (difluoromethoxy) pyridin-3-yl) -1, 1-diphenylmethane imine (2.3 g,5.70 mmol), N-methylpiperazine (685.2 mg,6.84 mmol) was added tris (dibenzylideneacetone) dipalladium (522.0 mg,0.57 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (468.0 mg,1.14 mmol) and cesium carbonate (5.6 g,17.19 mmol). The reaction was stirred under nitrogen at 90℃for 12 hours. After completion of the reaction, water (50.0 mL) was added thereto, followed by extraction with ethyl acetate (50 mL. Times.3). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (PE: ea=2:1) to give a brown oily liquid (310.0 mg). LC-MS: [ M+H ] ] + :423.3。
Synthesis of 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) pyridin-3-amine (5):
to a solution of N- (2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) pyridin-3-yl) -1, 1-diphenylmethanimine (310.0 mg,0.73 mmol) in dichloromethane (5.0 mL) was added trifluoroacetic acid (2.5 mL). The reaction system was stirred at room temperature for 2 hours. The reaction was concentrated and purified by silica gel column chromatography (DCM: meoh=10:1) to give a colorless oily liquid (80.0 mg). LC-MS: [ M+H ]] + :259.1。
Synthesis of 5' - (2- ((2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) pyridin-3-yl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-139):
to 2- (difluoromethoxy) -5- (4-methylpiperazin-1-yl) pyridin-3-amine (100.0 mg,0.39 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a suspension of 3' -ketone (common int-10) (123.0 mg,0.43 mmol) in dioxane (3.0 mL) was added palladium acetate (9.0 mg,0.04 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (48.0 mg,0.08 mmol) and cesium carbonate (252.0 mg,0.77 mmol). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction mixture was diluted with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give a crude product. The crude product was purified by preparative HPLC to give a yellow solid (17.0 mg). 1 H NMR(400MHz,DMSO)δ8.93(s,1H),8.81(d,J＝3.2Hz,1H),8.51(d,J＝2.8Hz,1H),8.41(s,1H),8.32(s,1H),8.26(d,J＝8.0Hz,1H),8.01(t,J＝59.6Hz,1H),7.52(d,J＝8.0Hz,1H),6.62(d,J＝2.4Hz,1H),2.96(t,J＝4.8Hz,4H),2.47(t,J＝4.8Hz,4H),2.22(s,3H),1.56(s,4H)；LC-MS:[M+H] + :512.2。
EXAMPLE 40 Synthesis of Compounds DP-01-150
Synthesis of 4-bromo-2-iodo-1-vinylbenzene (2):
methyl triphenylphosphine iodide (468.0 mg,1.16 mmol) and potassium carbonate (200.0 mg,1.45 mmol) were dissolved in dioxane (10 mL), and 4-bromo-2-iodo-benzaldehyde (300.0 mg,0.97 mmol) was added thereto and reacted at 100℃for 18 hours. The reaction solution was cooled, dried by spinning, and the crude product was subjected to silica gel column chromatography (petroleum ether) to give a colorless oil (230.0 mg). 1 H NMR(400MHz,DMSO)δ8.20(s,1H),7.92(d,J＝8.4Hz,1H),7.77(d,J＝8.4Hz,1H),6.92(dd,J＝17.2,11.1Hz,1H),5.97(d,J＝17.3Hz,1H),5.57(d,J＝11.1Hz,1H)。
Synthesis of 4-bromo-1- (2, 2-difluorocyclopropyl) -2-iodobenzene (3):
sodium iodide (148.0 mg,0.99 mmol) and 4-bromo-2-iodo-1-vinylbenzene (700.0 mg,2.27 mmol) were dissolved in tetrahydrofuran (3 mL), and (trifluoromethyl) trimethylsilane (1.13 g,7.95 mmol) was added thereto, and the reaction was cooled down at 70℃under nitrogen atmosphere. The reaction solution was cooled, dried by spinning, and the crude product was subjected to silica gel column chromatography (petroleum ether) to give a colorless oil (430.0 mg). 1 H NMR(400MHz,DMSO)δ8.10(s,1H),7.60(d,J＝8.0Hz,1H),7.24(d,J＝8.0Hz,1H),2.91(dd,J＝21.2,11.2Hz,1H),2.02–1.94(m,2H)。
Synthesis of N- (5-bromo-2- (2, 2-difluorocyclopropyl) phenyl) -1, 1-diphenylmethane imine (4):
4-bromo-1- (2, 2-difluorocyclopropyl) -2-iodobenzene (430.0 mg,1.20 mmol), benzophenone imine (218.0 mg,1.20 mmol), sodium t-butoxide (345.4 mg,3.59 mmol), tris (dibenzylideneacetone) dipalladium (109.7 mg,0.12 mmol) and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (138.6 mg,0.24 mmol) were dissolved in toluene (6 mL) and reacted at 60℃for 5 hours. The reaction solution was cooled, dried by spinning, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=10:1) to give a white solid (370.0 mg). LC-MS: [ M+H ] ] + :412.0,414.0。
Synthesis of N- (2, 2-difluorocyclopropyl) -5- (4-methylpiperazin-1-yl) phenyl) -1, 1-diphenylmethanimine (5):
n- (5-bromo-2- (2, 2-difluorocyclopropyl) phenyl) -1, 1-diphenylmethanimine (350.0 mg,0.85 mmol), N-methylpiperazine (102.1 mg,1.02 mmol), sodium t-butoxide (244.8 mg,2.55 mmol), tris (dibenzylideneacetone) dipalladium (77.7 mg,0.09 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (105.7 mg,0.17 mmol) were dissolved in toluene (8 mL) and reacted at 110℃for 3 hours. The reaction solution was cooled, dried by spinning, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol=10:1) to give a brown solid (350.0 mg). LC-MS: [ M+H ]] + :432.7。
Synthesis of 2- (2, 2-difluorocyclopropyl) -5- (4-methylpiperazin-1-yl) aniline (6):
n- (2, 2-Difluorocyclopropyl) -5- (4-methylpiperazin-1-yl) phenyl) -1, 1-diphenylmethanimine (100.0 mg,0.23 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (1 mL) was added dropwise, and the mixture was reacted at room temperature for 18 hours. The reaction mixture was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol: ammonia water=100:10:1) to give a white solid (60.0 mg). LC-MS: [ M+H ]] + :268.3。
Synthesis of 5' - (2- ((2- (2, 2-difluorocyclopropyl) -5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-150):
2- (2, 2-difluorocyclopropyl) -5- (4-methylpiperazin-1-yl) aniline (60.0 mg,0.22 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (65.0 mg,0.22 mmol), tris (dibenzylideneacetone) dipalladium (21.0 mg,0.02 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (28.0 mg,0.045 mmol) and cesium carbonate (220.0 mg,0.68 mmol) were dissolved in toluene (5 mL) and reacted at 100℃for 18 hours. The reaction solution was filtered, dried by spinning, and the crude product was prepared by neutrality to give a white solid (18.8 mg). 1 H NMR(400MHz,MeOD)δ8.51(s,1H),8.41(d,J＝3.6Hz,1H),8.33(d,J＝8.0Hz,1H),7.54(d,J＝2.4Hz,1H),7.37(d,J＝8.4Hz,1H),7.14(d,J＝8.4Hz,1H),6.76(dd,J＝8.8,2.8Hz,1H),3.24–3.21(m,4H),2.84–2.75(m,1H),2.62–2.59(m,4H),2.34(s,3H),1.83–1.80(m,1H),1.67–1.52(m,5H)；LC-MS:[M+H] + :521.8。
EXAMPLE 41 Synthesis of Compounds DP-01-149
Synthesis of 4-bromo-1- (bromomethyl) -2-iodobenzene (2):
the compound 4-bromo-2-iodo-1-toluene (9.0 g,30.31 mmol), N-bromosuccinimide (5.9 g,33.15 mmol) and azobisisobutyronitrile (0.5 g,3.04 mmol) were dissolved in carbon tetrachloride (50 mL) and reacted at 80℃for 2 hours. The reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-30%) to give a white solid (2.5 g). 1 H NMR(400MHz,DMSO-d6)δ8.09(d,J＝2.0Hz,1H),7.62(dd,J＝8.4,2.0Hz,1H),7.56(d,J＝8.4Hz,1H),4.70(s,2H)。
Synthesis of 4-bromo-2-iodo-1- (isocyanatomethyl) benzene (3):
to a solution of 4-bromo-1- (bromomethyl) -2-iodobenzene (2.4 g,6.39 mmol) in tetrahydrofuran (30 mL) was added trimethylcyanosilane (1.3 g,13.10 mmol). Then, the reaction mixture was cooled to 0℃and 1M tetrabutylammonium fluoride (12.8 mL,12.8 mmol) was slowly added dropwise, and after the completion of the addition, the reaction was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was quenched with ice-water mixture (30 mL) and the organic phase separated. The aqueous phase was extracted with ethyl acetate (30 mL. Times.2), and the organic phases were combined, and water (30 mL) and saturated brine were used, respectively (30 mL) was washed, dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=10:1) to give a white solid (1.5 g). 1 HNMR(400MHz,DMSO-d6)δ8.13(d,J＝1.4Hz,1H),7.67(dd,J＝8.4,1.4Hz,1H),7.47(d,J＝8.4Hz,1H),4.03(s,2H)。
Synthesis of 1- (4-bromo-2-iodophenyl) cyclopropane-1-carbonitrile (4):
4-bromo-2-iodo-1- (isocyanatomethyl) benzene (1.6 g,4.97 mmol) was dissolved in N, N-dimethylformamide (2 mL) under nitrogen, cooled to 0deg.C, and sodium hydride (398.4 mg,9.94mmol,60% w/w) was added. After stirring the mixture at 0℃for 1 hour, 1-chloro-2-bromoethane (0.82 mL,9.94 mmol) was added and stirring was continued for 1 hour after heating to 25 ℃. The reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=1:5) to give a white solid (1.5 g). 1 H NMR(400MHz,DMSO-d6)δ8.15(d,J＝2.5Hz,1H),7.64(dd,J＝9.0,2.5Hz,1H),7.39(d,J＝9.0Hz,1H),1.80(q,J＝5.1Hz,2H),1.38(q,J＝5.0Hz,2H)。
Synthesis of 1- (4-bromo-2- ((diphenylmethylene) amino) phenyl) cyclopropane-1-carbonitrile (5):
to a solution of 1- (4-bromo-2-iodophenyl) cyclopropane-1-carbonitrile (500.0 mg,1.44 mmol) and diphenylmethane imine (312.5 mg,1.72 mmol) in 1, 4-dioxane (10 mL) was added sequentially 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (178.9 mg,0.29 mmol), tris (dibenzylideneacetone) dipalladium (263.2 mg,0.29 mmol) and cesium carbonate (1.4 g,4.31 mmol) and stirred at 100℃for 2 hours under nitrogen. The reaction solution was cooled to room temperature and concentrated to dryness. The crude product was purified by silica gel column chromatography (methanol: dichloromethane=0-5%) to give a yellow solid (550.0 mg). LC-MS: [ M+H ] ] + :401.1。
Synthesis of 1- (2- ((diphenylmethylene) amino) -4- (4-methylpiperazin-1-yl) phenyl) cyclopropane-1-carbonitrile (6):
to 1- (4-bromo-2- ((diphenylmethylene) amino) phenyl) cyclopropane-1-carbonitrile (550 mg,1.37 mmol), 1-methylpiperazine (206.3 mg,2.06 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy under nitrogenTo a solution of 1, 4-dioxane (20 mL) in 1,1' -biphenol (56.3 mg,0.14 mmol) and tris (dibenzylideneacetone) dipalladium (125.6 mg,0.14 mmol) was added cesium carbonate (1340.1 mg,4.11mmol. The mixture was stirred at 100℃for 2 hours until complete consumption of the starting material, the reaction solution was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-5% methanol/dichloromethane) to give a yellow solid (500.0 mg.) LC-MS: [ M+H] + :421.1。
Synthesis of 1- (2-amino-4- (4-methylpiperazin-1-yl) phenyl) cyclopropane-1-carbonitrile (7):
to a solution of 1- (2- ((diphenylmethylene) amino) -4- (4-methylpiperazin-1-yl) phenyl) cyclopropane-1-carbonitrile (350.0 mg,0.83 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1.5 mL) at room temperature, the reaction solution was stirred at room temperature for 2 hours, then concentrated under reduced pressure, and the obtained residue was neutralized with saturated aqueous sodium hydrogencarbonate (5 mL) and extracted with dichloromethane (10 mL. Times.2). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated by filtration to give the crude product (150.0 mg) as a brown oil. The crude product was used directly in the next reaction. LC-MS: [ M+H ] ] + :257.3.1- (2- ((5-fluoro-4- (3 '-oxospiro [ cyclopropane-1, 1' -isoindoline))]-synthesis of 5' -yl) pyrimidin-2-yl) amino) -4- (4-methylpiperazin-1-yl) phenyl) cyclopropane-1-carbonitrile (DP-01-149):
to 1- (2-amino-4- (4-methylpiperazin-1-yl) phenyl) cyclopropane-1-carbonitrile (200.0 mg,0.78 mmol) and 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a solution of 3' -ketone (common int-10) (226.0 mg,0.78 mmol) in 1, 4-dioxane (10 mL) was added sequentially 1,1' -binaphthyl-2, 2' -bisdiphenylphosphine (48.6 mg,0.08 mmol), tris (dibenzylideneacetone) dipalladium (71.4 mg,0.08 mmol) and cesium carbonate (762.0 mg,2.34 mmol). The mixture was stirred at 100℃for 2 hours under nitrogen. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated by filtration to give the crude product. The crude product was isolated and purified by preparative HPLC to give a yellow solid (8.9 mg). 1 H NMR(400MHz,DMSO-d6)δ9.65(s,1H),9.06(d,J＝3.1Hz,1H),8.95(s,1H),8.35(s,1H),8.28(d,J＝8.2Hz,1H),7.98(d,J＝1.8Hz,1H),7.56(d,J＝8.0Hz,1H),6.88(d,J＝8.3Hz,1H),6.70(d,J＝8.1Hz,1H),3.14–3.05(m,4H),2.43(d,J＝4.8Hz,4H),2.20(s,3H),1.57(s,4H),1.47(d,J＝12.8Hz,4H)；LC-MS:[M+H] + :510.3。
EXAMPLE 42 Synthesis of Compounds DP-01-164
Synthesis of 1-methyl-4- (3-nitro-4- (trifluoromethyl) phenyl) piperazine (2):
4-bromo-2-nitrotrifluoromethylbenzene (1.0 g,3.70 mmol) was dissolved in toluene (10 mL) at room temperature, and 1-methylpiperazine (556.5 mg,5.56 mmol), tris (dibenzylideneacetone) dipalladium (169.6 mg,0.19 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (152.1 mg,0.37 mmol), and sodium t-butoxide (1.1 g,11.45 mmol) were added sequentially. The reaction mixture was stirred at 90℃for 6 hours, cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (15 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was chromatographed on a silica gel column (ethyl acetate/petroleum ether=1:1; r f Purified to give a colorless transparent oil (800.0 mg) =0.3). LC-MS: [ M+H ]] + :290.1。
Synthesis of 5- (4-methylpiperazin-1-yl) -2- (trifluoromethyl) aniline (3):
1-methyl-4- (3-nitro-4- (trifluoromethyl) phenyl) piperazine (40.0 mg,0.14 mmol) was dissolved in methanol (3 mL), pd/C (14.9 mg,10% w/w) was added and the hydrogen was replaced three times. The mixture was stirred at room temperature for 6 hours until the starting material had completely disappeared. The reaction solution was filtered through celite, and the filtrate was concentrated to give a black solid (30.0 mg). The crude product obtained was used directly in the next reaction. LC-MS: [ M+H ]] + :260.1。
Synthesis of 5' - (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethyl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-164):
5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (25.0 mg,0.09 mmol) and 5- (4-methylpiperazine-1-)To the solution was added palladium acetate (2.24 mg,0.01 mmol), cesium carbonate (84.4 mg,0.26 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (10.8 mg,0.017 mmol). The mixture was heated to reflux for 6 hours after three nitrogen substitutions. The reaction solution was poured into water (1 mL) and extracted with ethyl acetate (4 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by basic prep HPLC to give a clear colorless solid (3.2 mg). 1 H NMR(400MHz,DMSO-d6)δ8.85(s,1H),8.75(s,1H),8.56(d,J＝3.6Hz,1H),8.25(s,1H),8.15(d,J＝8.0Hz,1H),7.49(d,J＝8.8Hz,1H),7.45(d,J＝8.8Hz,1H),7.27(s,1H),6.89(d,J＝8.4Hz,1H),3.28–3.22(m,4H),2.46–2.36(m,4H),2.20(s,3H),1.52(s,4H)；LC-MS:[M+H] + :513.3。
EXAMPLE 43 Synthesis of Compounds DP-01-206
Synthesis of 1- (4-methoxy-3-nitrophenyl) -4-methylpiperazine (2):
the compound 4-bromo-1-methoxy-2-nitrobenzene (500.0 mg,2.16 mmol), 1-methylpiperazine (280.6 mg,2.80 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (88.5 mg,0.22 mmol), cesium carbonate (1404.2 mg,4.31 mmol) and tris (dibenzylideneacetone) dipalladium (98.7 mg,0.11 mmol) were dissolved in dioxane (8 mL), replaced with nitrogen three times, warmed to 90 ℃ and stirred for 6 hours, and LCMS detected complete reaction of the starting materials. The reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-34%, petroleum ether/ethyl acetate=1:1, rf=0.2) to give a yellow oil (230.0 mg). LC-MS: [ M+H ]] + :252.1。
Synthesis of 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (3):
the compound 1- (4-methoxy-3-nitrophenyl) -4-methylpiperazine (180.0 mg,0.72 mmol) and Pd/C (80 mg,10% w/w) were suspended in methanol5 mL), the reaction was stirred at room temperature for 4 hours after three hydrogen substitutions. After completion of the LCMS monitoring reaction, the reaction was filtered through celite and the filter cake was washed 3 times with ethyl acetate to give a yellow solid (150.0 mg) after the filtrate was spun dry. LC-MS: [ M+H ] ] + :222.2。
Synthesis of 5' - (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-206):
the compound 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (150.0 mg,0.68 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (216.0 mg,0.75 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (84.4 mg,0.14 mmol), palladium acetate (15.2 mg,0.07 mmol) and cesium carbonate (441.7 mg,1.36 mmol) were dissolved in 1, 4-dioxane (4 mL), nitrogen was replaced, and stirring was carried out at 100℃for 6 hours, and the disappearance of the starting material was monitored by LCMS. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (methanol: dichloromethane=0-16%; dichloromethane/methanol=10:1, r f Crude black solid (111.0 mg) was purified by prep HPLC to give a white solid (27.3 mg). 1 H NMR(400MHz,CDCl 3 )δ8.65(s,1H),8.44–8.33(m,3H),7.87(s,1H),7.33(s,1H),7.18(d,J＝8.4Hz,1H),6.83(d,J＝8.8Hz,1H),6.56(dd,J＝8.4,2.8Hz,1H),3.89(s,3H),3.29–3.10(m,4H),2.66–2.56(m,4H),2.37(s,3H),1.70–1.60(m,2H),1.56-1.52(m,2H)；LC-MS:[M+H] + :475.3。
EXAMPLE 44 Synthesis of Compound DP-01-221
Synthesis of 4-bromo-1-ethoxy-2-nitrobenzene (2):
4-bromo-1-fluoro-2-nitrobenzene (1.0 g,4.55 mmol) was dissolved in N, N-dimethylformamide (10 mL) at room temperature, and sodium ethoxide (619.3 mg,9.1 mmol) was added to the solution. The mixture was stirred at 50℃for 12 hours, Until the raw materials are completely consumed. The reaction mixture was cooled, quenched with water (10 mL), and extracted with ethyl acetate (50 mL. Times.3). The organic phases were combined, washed with saturated brine (15 mL. Times.3), finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=3:1) afforded a white solid (1.0 g). LC-MS: [ M+H ]] + :246.2。
Synthesis of 1- (4-ethoxy-3-nitrophenyl) -4-methylpiperazine (3):
to a solution of 4-bromo-1-ethoxy-2-nitrobenzene (1.0 g,4.08 mmol) in dioxane (10 mL) was added sequentially 1-methylpiperazine (529.2 mg,5.28 mmol), tris (dibenzylideneacetone) dipalladium (186.1 mg,0.20 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (166.9 mg,0.41 mmol) and cesium carbonate (2648.3 mg,8.13 mmol). The resulting mixture was stirred under nitrogen at 100 ℃ for 6 hours. After the reaction mixture was cooled, it was diluted with water (10 mL) and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=3:1) afforded a white solid (400.0 mg). LC-MS: [ M+H ]] + :266.2。
Synthesis of 2-ethoxy-5- (4-methylpiperazin-1-yl) aniline (4):
1- (4-ethoxy-3-nitrophenyl) -4-methylpiperazine (200.0 mg,0.75 mmol) and palladium on carbon (80.22 mg,10% w/w) were dissolved in methanol (3 mL). A hydrogen balloon was inserted and replaced three times with hydrogen. The mixture was stirred at room temperature for 5 hours until the starting material was completely consumed. The reaction solution was concentrated by filtration to give a black solid (120.0 mg). LC-MS: [ M+H ] ] + :236.2。
Synthesis of 5' - (2- ((2-ethoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-221):
to 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline at room temperature]To a solution of 3' -ketone (common int-10) (135.4 mg,0.47 mmol) and 2-ethoxy-5- (4-methylpiperazin-1-yl) aniline (100.0 mg,0.43 mmol) in dioxane (2 mL) was added cesium carbonate (415.4 mg,1.28 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (52.9 mg,0.085 mmol) and tris (dibenzylideneacetone) dipalladium (389mg,0.043 mmol). The reaction solution was heated under reflux for 6 hours under nitrogen protection until the starting material was completely consumed. The reaction mixture was cooled, the solvent was removed in vacuo, diluted with water (5 mL), and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Basic preparation gave a white solid (20.7 mg). 1 H NMR(400MHz,DMSO-d6)δ8.64(s,1H),8.48–8.28(m,3H),7.88(s,1H),7.18(d,J＝8.0Hz,1H),6.82(d,J＝8.8Hz,1H),6.54(dd,J＝8.8,2.8Hz,1H),6.45(s,1H),4.10(q,J＝7.2Hz,2H),3.28–3.09(m,4H),2.65–2.52(m,4H),2.37(s,3H),1.65(m,2H),1.54(m,2H),1.47(t,J＝6.8Hz,3H)；LC-MS:[M+H] + :489.2。
EXAMPLE 45 Synthesis of Compounds DP-01-207
Synthesis of 4-bromo-1-isopropoxy-2-nitrobenzene (2):
the compound 4-bromo-2-nitrophenol (2.0 g,9.17 mmol) and 2-iodopropane (1.1 mL,11.01 mmol) were dissolved in N, N-dimethylformamide (20.0 mL), and potassium carbonate (1.9 g,13.74 mmol) was added to the solution. The mixture was stirred at 110℃for 6 hours. After the reaction was completed, the reaction mixture was cooled, quenched with water (20 mL), and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=3:1) afforded a white solid (1.8 g). LC-MS: [ M+H ] ] + :260.1,262.1。
Synthesis of 1- (4-isopropoxy-3-nitrophenyl) -4-methylpiperazine (3):
4-bromo-1-isopropoxy-2-nitrobenzene (1.0 g,3.83 mmol) and N-methylpiperazine (501.0 mg,5.00 mmol) were dissolved in dioxane (10 mL), and to the solution were added tris (dibenzylideneacetone) dipalladium (176.0 mg,0.19 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (157.9 mg,0.38 mmol) and cesium carbonate (2505.4 mg,7.69 mmol). The resulting mixture was stirred at 100deg.C for 6 hours, after the reaction was completed, the reaction mixture was cooled, quenched with water (10 mL), and quenched with acetic acidEthyl ester extraction (10 ml×3). The combined organic phases were washed with saturated brine (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether/ethyl acetate=3:1) afforded a white solid (700.0 mg). LC-MS: [ M+H ]] + :280.1。
Synthesis of 2-isopropoxy-5- (4-methylpiperazin-1-yl) aniline (4):
1- (4-isopropoxy-3-nitrophenyl) -4-methylpiperazine (300.0 mg,1.07 mmol) and palladium on carbon (114.3 mg,10% w/w) were dissolved in methanol (3 mL). A hydrogen balloon was inserted and replaced three times with hydrogen. The mixture was stirred for 5 hours until the starting material was completely consumed. The reaction solution was concentrated by filtration to give a black solid (200.0 mg). LC-MS: [ M+H ]] + :250.1。
Synthesis of 5' - (5-fluoro-2- ((2-isopropoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-207):
5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (100.0 mg,0.35 mmol) and 2-isopropoxy-5- (4-methylpiperazin-1-yl) aniline (111.9 mg,0.45 mmol) were dissolved in dioxane (1 mL), tris (dibenzylideneacetone) dipalladium (31.61 mg,0.035 mmol) was added to the solution, followed by cesium carbonate (337.4 mg,1.04 mmol) and 1,1' -binaphthyl-2, 2' -bisdiphenylphosphine (43.0 mg,0.07 mmol) were replaced with nitrogen three times. The mixture was refluxed for 6 hours. After completion of the reaction, the solvent was removed in vacuo, diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The combined organic phases were washed with saturated brine (5.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by prep. to give a white solid (39.2 mg). 1 H NMR(400MHz,DMSO-d6)δ8.90(s,1H),8.67(d,J＝3.5Hz,1H),8.30(s,1H),8.24(d,J＝8.3Hz,1H),8.16(s,1H),8.06(s,1H),8.03(d,J＝2.8Hz,1H),7.49(d,J＝8.1Hz,1H),6.95(d,J＝8.9Hz,1H),6.56(dd,J＝8.9,2.8Hz,1H),4.53–4.47(m,1H),3.08–3.05(m,4H),2.47(d,J＝4.6Hz,4H),2.22(s,3H),1.55(s,4H),1.28(d,J＝6.0Hz,6H)；LC-MS:[M+H] + :503.2。
EXAMPLE 46 Synthesis of Compound PR-01-116
Synthesis of 5' - (5-fluoro-2- ((5- (1-methyl-4-oxo-1, 4-azaphosphin-4-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (PR-01-116):
the compound 1-methyl-1, 4-azaphosphine-4-oxide (40.0 mg,0.30 mmol), 5'- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ]3' -Ketone (common int-12) (152.8 mg,0.30 mmol), tris (dibenzylideneacetone) dipalladium (27.5 mg,0.03 mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (17.4 mg,0.03 mmol), potassium phosphate (127.2 mg,0.6 mmol) were dissolved in N, N-dimethylformamide (2 mL) and reacted at 100℃for 4 hours under nitrogen protection. The reaction solution was cooled and filtered, the filtrate was diluted with water (30 mL), extracted with ethyl acetate (30 mL. Times.3), and concentrated to a black-like oil, which was purified by preparative HPLC to give a white solid (55 mg). 1 HNMR(400MHz,MeOD-d4)δ8.94–8.90(m,1H),8.56(d,J＝3.6Hz,1H),8.51(s,1H),8.40–8.38(m,1H),7.58–7.56(m,2H),7.45(d,J＝8.4Hz,1H),3.05–2.94(m,2H),2.90–2.80(m,2H),2.38–2.29(m,5H),2.17–2.08(m,2H),1.71–1.58(m,4H)；LC-MS:[M+H] + :562.2。
EXAMPLE 47 Synthesis of Compounds DP-01-160
Synthesis of 5' - (5-fluoro-2- ((5- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (1):
to 5'- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a mixed solvent of 3 '-ketone (common int-12) (100.0 mg,0.20 mmol), 1-methyl-4-boronic acid pinacol ester-1, 2,3, 6-tetrahydropyridine (56.9 mg,0.26 mmol) 1, 4-dioxane (1.0 mL) and water (0.1 mL) were added 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride (14.4 mg,0.02 mmol) and potassium phosphate(124.8 mg,0.59 mmol). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched by addition of an aqueous solution (10.0 mL) and extracted three times with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was isolated as a yellow solid (8.8 mg). 1 H NMR(400MHz,DMSO-d6)δ9.30(s,1H),8.91(s,1H),8.65(d,J＝3.6Hz,1H),8.30(s,1H),8.20(d,J＝8.4Hz,1H),8.02(s,1H),7.49(d,J＝8.0Hz,1H),7.36(d,J＝8.4Hz,1H),7.29(d,J＝8.4Hz,1H),6.21(s,1H),3.14(s,2H),2.69(s,2H),2.53(s,2H),2.36(s,3H),1.54(s,4H)；LC-MS:[M+H] + :526.4。
Synthesis of 5' - (5-fluoro-2- ((5- (1-methylpiperidin-4-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-160):
to 5'- (5-fluoro-2- ((5- (1-methyl-1, 2,3, 6-tetrahydropyridin-4-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a solution of 3' -ketone (20.0 mg,0.04 mmol) in methanol (2.0 mL) was added palladium on carbon (10.0 mg,10% w/w). The reaction system was stirred at room temperature for 24 hours under a hydrogen atmosphere. After the reaction was completed, palladium on carbon was removed by filtration and concentrated in vacuo. The crude product was isolated as a yellow solid (4.0 mg). 1 H NMR(400MHz,MeOD)δ8.66(s,1H),8.54(d,J＝4.0Hz,1H),8.52(d,J＝2.0Hz,1H),8.48(s,1H),8.36(d,J＝8.0Hz,1H),7.42(d,J＝8.0Hz,1H),7.33(dd,J＝8.4,1.2Hz,1H),7.05(dd,J＝8.4,2.0Hz,1H),3.60(d,J＝12.4Hz,2H),3.19–3.12(m,2H),3.04–2.95(m,1H),2.92(s,3H),2.21–2.06(m,4H),1.70–1.61(m,4H)；LC-MS:[M+H] + :528.3。
EXAMPLE 48 Synthesis of Compounds DP-01-163
Synthesis of 5' - (5-fluoro-2- ((5- ((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1] oct-3-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-163):
5'- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] at room temperature]-3' -Ketone (common int-12) (50.0 mg,0.10 mmol) was dissolved in tetrahydrofuran (0.5 mL) and 8-methyl-3, 8-diaza-bicyclo [3.2.1] was added]Octane hydrochloride (24.0 mg,0.12 mmol), 1M lithium bis (trimethylsilylamide) (0.49 mL,0.49 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (3.9 mg,0.01 mmol), and tris (dibenzylideneacetone) dipalladium (9.2 mg,0.01 mmol). The resulting mixture was stirred at 90℃for 5 hours, and was dried by filtration to give a blackish brown solid, which was subjected to basicity to give a white solid (8.2 mg). 1 H NMR(400MHz,CDCl 3 )δ8.61(s,1H),8.42(d,J＝3.2Hz,1H),8.36(d,J＝8.1Hz,1H),8.16(d,J＝2.9Hz,1H),7.44(s,1H),7.19(d,J＝8.1Hz,1H),7.13(d,J＝9.1Hz,1H),6.42(dd,J＝9.4,3.1Hz,2H),3.36(d,J＝10.9Hz,2H),3.27(s,2H),3.09(d,J＝10.2Hz,2H),2.37(s,3H),2.11–1.93(m,2H),1.77–1.54(m,6H)；LC-MS:[M+H] + :555.3。
EXAMPLE 49 Synthesis of Compounds DP-01-162
Synthesis of tert-butyl 6-methyl-3, 6-diazabicyclo [3.1.1] heptane-3-carboxylate (2):
3, 6-diazabicyclo [3.1.1]Heptane-3-carboxylic acid tert-butyl ester (400.0 mg,2.02 mmol) and formaldehyde (0.65 mL,8.07mmol,37% aqueous formaldehyde) were dissolved in tetrahydrofuran (10 mL) and acetic acid (0.5 mL), stirred at room temperature for 1 hour, sodium borohydride acetate (2.1 g,9.91 mmol) was added to the reaction system, and stirring was continued at room temperature for 17 hours. The reaction solution was diluted with water (8 mL), and the solution was adjusted to ph=9 with potassium carbonate and extracted with ethyl acetate (20 ml×3). The combined organic phases were washed with saturated aqueous sodium chloride (10 mL), dried over anhydrous sodium sulfate, and concentrated by filtration to give a white solid (400.0 mg). LC-MS: [ M+H ]] + :213.2。
Synthesis of 6-methyl-3, 6-diazabicyclo [3.1.1] heptane dihydrochloride (3):
6-methyl-3, 6-diazabicyclo [3.1.1]Heptane-3-carboxylic acid tert-butyl ester(270.0 mg,1.27 mmol) was dissolved in 4M hydrochloric acid (5 mL,20.00 mmol) in 1, 4-dioxane and stirred at room temperature for 6 hours. Methyl tert-butyl ether (15 mL) was added, the solid precipitated, filtered, and the methyl tert-butyl ether (20 mL) was washed, the solid collected and dried to give the product as a white solid (230.0 mg). LC-MS: [ M+H ] ] + :113.2。
Synthesis of 5' - (5-fluoro-2- ((5- (6-methyl-3, 6-diazabicyclo [3.1.1] heptan-3-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-162):
the compound 5'- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-3' -one (common int-12) (50.0 mg,0.10 mmol), 6-methyl-3, 6-diazabicyclo [3.1.1]]Heptanedihydrochloride (21.8 mg,0.19 mmol), tris (dibenzylideneacetone) dipalladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M lithium bis trimethylsilylamide (0.49 mL,0.49 mmol) were dissolved in tetrahydrofuran (0.5 mL) and reacted at 70℃for 6 hours under nitrogen. The reaction solution was cooled to room temperature, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (10 mL. Times.3). The organic phases are combined and dried, filtered and spun-dried. The crude product was prepared from reverse phase formic acid to give a white solid (7.2 mg). 1 H NMR(400MHz,DMSO-d6)δ8.97(s,1H),8.87(s,1H),8.65(d,J＝3.6Hz,1H),8.32(s,1H),8.21(s,1H),8.19(s,1H),7.47–7.45(m,2H),7.23(d,J＝8.0Hz,1H),6.51(dd,J＝8.8,2.8Hz,1H),3.60(d,J＝5.6Hz,2H),3.48(d,J＝10.8Hz,2H),3.31–3.28(m,2H),2.46–2.40(m,1H),2.04(s,3H),1.56–1.53(m,5H)；LC-MS:[M+H] + :541.4。
EXAMPLE 50 Synthesis of Compounds PR-01-113, PR-01-113A, PR-01-113B
Synthesis of tert-butyl 5- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0] heptane-2-carboxylate (2):
2, 5-diazabicyclo [4.1.0] compound ]Tert-butyl heptane-2-carboxylate (250.0 mg,1.26 mmol), 4-bromo-2-nitrotrifluoromethoxybenzene (360.6 mg,1.26 mmol), tris (dibenzylideneacetone) palladium (115.5 mg,0.126 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (120.2 mg,0.25 mmol) and sodium tert-butoxide (363.5 mg,3.78 mmol) were dissolved in toluene (8 mL) and reacted at 90℃for 2 hours under nitrogen. The reaction solution was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=3:1) to give a brown solid (210.0 mg). LC-MS: [ M ] t Bu+H] + :348.0。
Synthesis of 2- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0] heptane dihydrochloride (3):
5- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0]Tert-butyl heptane-2-carboxylate (210.0 mg,0.52 mmol) was dissolved in 2N HCl (10 mL,20.00 mmol) in ethyl acetate and reacted at room temperature for 18 h. The reaction solution was dried by spin to give a yellow oily substance (195.0 mg). LC-MS: [ M+H ]] + :304.2。
Synthesis of 2-methyl-5- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0] heptane (4):
2- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0]Heptane dihydrochloride (190.0 mg,0.63 mmol) and 37% formaldehyde (0.27 mL,2.70 mmol) in water were dissolved in tetrahydrofuran (10 mL) and acetic acid (0.5 mL), stirred at room temperature for 1 hour, and sodium borohydride acetate (532.7 mg,2.51 mmol) was added to the reaction system and reacted at room temperature for 17 hours. Dilution with water (8 mL), potassium carbonate solids adjusted ph=9, extraction with ethyl acetate (10 ml×3), washing of the organic phase with saturated aqueous sodium chloride (10 mL), drying, spin-drying gave the crude product as a brown oily substance (140.0 mg) by column chromatography on silica gel (dichloromethane: methanol=10:1). LC-MS: [ M+H ] ] + :318.2。
Synthesis of 5- (5-methyl-2, 5-diazabicyclo [4.1.0] heptane-2-yl) -2- (trifluoromethoxy) aniline (5):
2-methyl-5- (3-nitro-4- (trifluoromethoxy) phenyl) -2, 5-diazabicyclo [4.1.0]Heptane (130.0 mg,0.41 mmol) was dissolved in methanol (10 mL), seven drops of concentrated hydrochloric acid and 10% palladium on carbon (15.0 mg) were added, and the mixture was reacted at room temperature under hydrogen protection for 2 hours. Filtering, spin-drying the filtrate to obtain brown solid, adding saturated sodium bicarbonate waterThe solution (10 mL) was extracted with ethyl acetate (10 mL. Times.3), and the organic phase was dried and spun-dried to give a brown solid (115.0 mg). LC-MS: [ M+H ]] + :288.3。
Synthesis of 5' - (5-fluoro-2- ((5- (5-methyl-2, 5-diazabicyclo [4.1.0] heptan-2-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (PR-01-113):
the compound 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-3' -one (141.2 mg,0.49 mmol), 5- (5-methyl-2, 5-diazabicyclo [ 4.1.0)]Heptan-2-yl) -2- (trifluoromethoxy) aniline (common int-10) (100.0 mg,0.35 mmol), tris (dibenzylideneacetone) palladium (31.9 mg,0.035 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (43.4 mg,0.07 mmol) and cesium carbonate (340.2 mg,1.04 mmol) were dissolved in toluene (1 mL) and reacted at 100℃for 5 hours under nitrogen protection. The reaction solution was filtered, and the filtrate was dried by spin. The crude product was prepared from formic acid to give a white solid (79.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.87(s,1H),8.63(d,J＝3.6Hz,1H),8.29(s,1H),8.19(d,J＝8.0Hz,1H),8.13(s,1H),7.50–7.46(m,2H),7.20(d,J＝8.4Hz,1H),6.61(dd,J＝9.2,2.8Hz,1H),3.36(s,1H),3.10(t,J＝8.4Hz,1H),2.64–2.61(m,2H),2.52(s,1H),2.40–2.32(m,4H),1.53(s,4H),0.58(d,J＝6.0Hz,1H),0.37(s,1H).LC-MS:[M+H] + :541.2。
The racemate PR-01-113 was purified by SFC separation (column: chiralPak AD, 250X 30mm I.D.,5 μm; mobile phase: A phase: CO) 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: IPA (0.1% 7mol/L NH3 in MeOH); gradient: b%:30%;7min,120 min) to give PR-01-113A (13.0 mg) as a white solid and PR-01-113B (17.5 mg) as a white solid.
PR-01-113A：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: IPA (0.05% DEA v/v); gradient: b%:30.0% retention time of 4.477min, ee% = 100%.
1 H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.87(s,1H),8.63(d,J＝3.2Hz,1H),8.29(s,1H),8.19(d,J＝8.0Hz,1H),7.50–7.46(m,2H),7.20(d,J＝8.4Hz,1H),6.61(dd,J＝9.2,3.2Hz,1H),3.35(s,1H),3.09(t,J＝8.4,1H),2.67–2.59(m,2H),2.48–2.44(m,1H),2.37–2.32(m,4H),1.53(s,4H),0.56–0.53(m,1H),0.33–0.31(m,1H)；LC-MS:[M+H] + :541.1。
PR-01-113B：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: IPA (0.05% DEA v/v); gradient: b%:30.0% retention time 5.334min, ee% = 97.37%.
1 H NMR(400MHz,DMSO-d6)δ8.98(s,1H),8.87(s,1H),8.63(d,J＝3.6Hz,1H),8.29(s,1H),8.19(d,J＝8.4Hz,1H),7.50–7.46(m,2H),7.20(d,J＝8.0Hz,1H),6.61(dd,J＝8.8,2.8Hz,1H),3.35(s,1H),3.09(t,J＝8.4,1H),2.67–2.59(m,2H),2.48–2.44(m,1H),2.37–2.30(m,4H),1.53(s,4H),0.55(q,J＝4.8Hz,1H),0.32(q,J＝4.8Hz,1H)；LC-MS:[M+H] + :541.2。
EXAMPLE 51 Synthesis of Compounds DP-01-203, DP-01-203A, DP-01-203B
Synthesis of 5' - (5-fluoro-2- ((5- (5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-203):
the compound 5'- (2- ((5-bromo-2- (trifluoromethoxy) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-3' -one (common int-12) (50.0 mg,0.10 mmol), 2-methyl-2, 5-diazabicyclo [ 2.2.1) ]Heptanedihydride (18.2 mg,0.10 mmol), tris (dibenzylideneacetone) dipalladium (9.0 mg,0.01 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (7.7 mg,0.02 mmol) and 1M lithium bis trimethylsilylamide (0.5 mL,0.50 mmol) were dissolved in tetrahydrofuran (0.5 mL) and reacted at 70℃for 4 hours under nitrogen. The reaction was cooled, quenched with saturated aqueous ammonium chloride (1 mL) and extracted with ethyl acetate (2 mL). The organic phase was dried, spun-dried, and the crude product was purified by column chromatography on silica gel (dichloromethane: methanol=7:1) to give crude product, which was prepared from reversed-phase formic acid to give a white solid(34.3mg)。 1 H NMR(400MHz,DMSO-d6)δ8.94(s,1H),8.87(s,1H),8.63(d,J＝3.2Hz,1H),8.29(s,1H),8.19–8.18(m,2H),7.47(d,J＝8.4Hz,1H),7.23(d,J＝2.8Hz,1H),7.14(d,J＝8.8Hz,1H),6.36(dd,J＝8.8,2.8Hz,1H),4.26(s,1H),3.47(s,1H),3.38–3.35(s,1H),3.19(d,J＝9.2Hz,1H),2.76(d,J＝7.6Hz,1H),2.54(s,1H),2.27(s,3H),1.88(d,J＝8.8Hz,1H),1.79(d,J＝8.8Hz,1H),1.53(s,4H)；LC-MS:[M+H] + :541.2。
The racemate DP-01-203 was purified by SFC separation (column: chiralPak C-IG, 250X 30mm I.D.,5 μm; mobile phase: A phase: CO) 2 ；B：MeOH+0.1％NH 3 -H 2 O; gradient: b%:40%;8.4min;180 min) to give white solid DP-01-203A (6.4 mg) and white solid DP-01-203B (7.7 mg).
DP-01-203A：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH (0.05% DEA v/v); gradient: b%:40.0% retention time of 3.562min, ee% = 99.77%.
1 H NMR(400MHz,DMSO-d6)δ8.93(s,1H),8.87(s,1H),8.63(d,J＝3.6Hz,1H),8.33(s,1H),8.29(s,1H),8.19(d,J＝8.0Hz,1H),7.47(d,J＝8.0Hz,1H),7.23(d,J＝2.8Hz,1H),7.13(d,J＝8.8Hz,1H),6.35(dd,J＝8.8,2.8Hz,1H),4.23(d,J＝8.4Hz,1H),3.42(s,1H),3.36(s,1H),3.17(d,J＝8.8Hz,1H),2.74(d,J＝8.0Hz,1H),2.46(s,1H),2.24(s,3H),1.85(d,J＝9.2Hz,1H),1.77(d,J＝9.2Hz,1H),1.53(s,4H)；LC-MS:[M+H] + :541.1。
DP-01-203B：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH (0.05% DEAVV/v); gradient: b%:40.0% retention time of 4.433 min, ee% = 90.05%.
1 H NMR(400MHz,DMSO-d6)δ8.93(s,1H),8.87(s,1H),8.63(d,J＝3.2Hz,1H),8.29(s,1H),8.19(d,J＝8.4Hz,1H),7.47(d,J＝8.0Hz,1H),7.23(d,J＝2.8Hz,1H),7.14(d,J＝8.0Hz,1H),6.35(dd,J＝9.2,2.8Hz,1H),4.24(s,1H),3.42(s,2H),3.17(d,J＝8.4Hz,1H),2.74(d,J＝7.6Hz,1H),2.46(s,1H),2.24(s,3H),1.85(d,J＝9.2Hz,1H),1.77(d,J＝9.6Hz,1H),1.53(s,4H)；LC-MS:[M+H] + :541.1。
EXAMPLE 52 Synthesis of Compounds DP-01-161, DP-01-161A, DP-01-161B
Synthesis of 3- (hydroxymethyl) -4-methylpiperazine-1-carboxylic acid tert-butyl ester (2):
the compound 3- (hydroxymethyl) piperazine-1-carboxylic acid tert-butyl ester (2.0 g,9.25 mmol) and formaldehyde (2.3 g,28.37mmol, 37%), glacial acetic acid (0.5 mL) were dissolved in methanol (20 mL) and stirred at room temperature for 1 hour, then cooled to 0 ℃, sodium cyanoborohydride (1.16 g,18.50 mmol) was added in portions and stirred at room temperature for 15 hours, and TLC monitored the starting material was complete. The reaction mixture was diluted with saturated sodium bicarbonate (40 mL) and extracted with dichloromethane (50 mL. Times.3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (methanol: dichloromethane=0-5%, dichloromethane/methanol=10:1, r f Purified to give a yellow oil (1.5 g) =0.3). LC-MS: [ M+H ]] + :231.1。
Synthesis of 3- (fluoromethyl) -4-methylpiperazine-1-carboxylic acid tert-butyl ester (3):
the compound 3- (hydroxymethyl) -4-methylpiperazine-1-carboxylic acid tert-butyl ester (1.5 g,6.51 mmol) was dissolved in dichloromethane (24 mL), a solution of diethylaminosulfur trifluoride (1.2 g,7.44 mmol) in dichloromethane (24 mL) was added dropwise to the above solution after cooling to 0deg.C, and stirring was continued for 12 hours at 40deg.C, and LCMS monitored the starting materials had reacted completely. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (20 mL) and extracted with dichloromethane (20 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (methanol: dichloromethane=0-3%; dichloromethane/methanol=20:1, r f Purified to give a colorless oil (600.0 mg) =0.4). LC-MS: [ M+H ]] + :233.1. Synthesis of 2- (fluoromethyl) -1-methylpiperazine (4):
the compound 3- (fluoromethyl) -4-methyl at room temperatureTert-butyl piperazine-1-carboxylate (500.0 mg,2.15 mmol) was dissolved in methanol (4 mL), a 2M hydrochloric acid/ethyl acetate solution (2.2 mL) was slowly added, the temperature was raised to 40℃and stirred for 6 hours, and LCMS detected complete reaction of the starting materials. The reaction solution was directly concentrated to give a crude product (250.0 mg) which was directly used in the next reaction. LC-MS: [ M+H ]] + :133.2。
Synthesis of 2- (fluoromethyl) -1-methyl-4- (3-nitro-4- (trifluoromethoxy) phenyl) piperazine (5):
the compound 2- (fluoromethyl) -1-methylpiperazine (250.0 mg,1.89 mmol), 4-bromo-2-nitro-1- (trifluoromethoxy) benzene (450.0 mg,1.57 mmol), tris (dibenzylideneacetone) dipalladium (72.0 mg,0.078 mmol), cesium carbonate (1.5 g,4.60 mmol) and 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (64.6 mg,0.16 mmol) were dissolved in 1, 4-dioxane (5 mL) under nitrogen and the mixture heated to 90 ℃ and stirred for 6 hours, LCMS detects complete reaction. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-70%, petroleum ether/ethyl acetate=1:1, r f Purified to give a yellow oil (120.0 mg) =0.3). LC-MS: [ M+H ]] + :338.1。
Synthesis of 5- (3- (fluoromethyl) -4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (6):
the compound 2- (fluoromethyl) -1-methyl-4- (3-nitro-4- (trifluoromethoxy) phenyl) piperazine (120.0 mg,0.36 mmol) and Pd/C (50 mg,10% w/w) were suspended in methanol (3 mL). A hydrogen balloon was inserted, and after three substitutions, the reaction was stirred at 40℃for 6 hours. LCMS monitored completion of the reaction, the reaction was filtered through celite, the filter cake was washed 3 times with ethyl acetate, and the filtrate was dried to give a white solid (100.0 mg, 92%). LC-MS: [ M+H ]] + :308.1。
Synthesis of 5' - (5-fluoro-2- ((5- (3- (fluoromethyl) -4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-161):
the compound 5- (3- (fluoromethyl) -4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (100.0 mg,0.33 mol), 5' - (2-)Chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (105.2 mg,0.36 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (41.1 mg,0.066 mmol), palladium acetate (7.4 mg,0.033 mmol) and cesium carbonate (322.6 mg,0.99 mmol) were dissolved in 1, 4-dioxane (5 mL) and the mixture was stirred for 6 hours after heating to 100deg.C, and LCMS monitored complete reaction of the starting materials. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by prep. to give a white solid (18.0 mg). 1 H NMR(400MHz,CDCl 3 )δ8.62(s,1H),8.45–8.29(m,3H),7.48(s,1H),7.26-7.15(m,2H),7.04(s,1H),6.56(dd,J＝8.8,2.8Hz,1H),4.64–4.35(m,2H),3.55(t,J＝13.2Hz,2H),3.04–2.95(m,2H),2.87(t,J＝6.8Hz,1H),2.58–2.55(m,2H),2.47(s,3H),1.67(t,J＝6.8Hz,2H),1.56(t,J＝6.8Hz,2H)；LC-MS:[M+H] + :561.3。
The racemate DP-01-161 was purified by SFC separation (column: chiralCel OX, 250X 21.2mm I.D.,5 μm; mobile phase: A phase: CO) 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: IPA+0.1% NH 3 -H 2 O; gradient: b%:30%,6.9min;180 min) to give white solid DP-01-161A (5.0 mg) and white solid DP-01-161B (7.0 mg).
DP-01-161A：
Chiral analysis conditions: chromatographic column: chiralCel OX,250×21.2mm i.d.,5 μm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH (0.05% DEA v/v); gradient: b:30% retention time of 3.938min; ee% = 100.0%.
1 H NMR(400MHz,CDCl 3 )δ8.62(s,1H),8.43(d,J＝3.2Hz,1H),8.36(d,J＝8.6Hz,1H),8.31(d,J＝2.8Hz,1H),7.47(s,1H),7.22(d,J＝8.0Hz,1H),7.18–7.15(m,1H),6.56(dd,J＝9.2,2.8Hz,1H),6.29(s,1H),4.54(d,J＝3.6Hz,1H),4.43–4.41(m,1H),3.55(t,J＝12.8Hz,2H),3.06–2.89(m,2H),2.88–2.79(m,1H),2.53–2.48(m,2H),2.43(s,3H),1.65–1.62(m,2H),1.56–1.54(m,2H)；LC-MS:[M+H] + :561.1。
DP-01-161B：
Chiral analysis conditions: chromatographic column: chiralCel OX,250×21.2mm I.D.,5 μm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH (0.05% DEA v/v); gradient: b:30% retention time 4.659min; ee% = 96.3%.
1 H NMR(400MHz,CDCl 3 )δ8.62(s,1H),8.43(d,J＝3.2Hz,1H),8.36(d,J＝8.0Hz,1H),8.31(d,J＝2.8Hz,1H),7.47(s,1H),7.22(d,J＝8.0Hz,1H),7.17(d,J＝8.8Hz,1H),6.56(dd,J＝9.2,3.2Hz,1H),6.33(s,1H),4.54(d,J＝4.0Hz,1H),4.3–4.41(m,1H),3.55(t,J＝12.8Hz,2H),3.04–2.97(m,1H),2.96–2.77(m,2H),2.53–2.47(m,2H),2.43(s,3H),1.65–1.61(m,2H),1.55–1.53(m,2H)；LC-MS:[M+H] + :561.3。
EXAMPLE 53 Synthesis of Compound DP-01-201-2
Synthesis of 5-bromo-7- (2-hydroxyethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (1):
5-bromo-1, 1-dimethyl-3-oxo-7-vinylisoindoline-2-carboxylic acid tert-butyl ester (300.0 mg,0.82 mmol) was dissolved in tetrahydrofuran (30 mL), 1M borane (1.0 mL) was added under ice bath to the solution, after reacting at room temperature for 1 hour, methanol (1 mL), sodium hydroxide (65.5 mg,1.64 mmol) and 30% hydrogen peroxide (1.17 mL) were added under ice bath, and after stirring at room temperature for 2 hours, the reaction was quenched with saturated aqueous sodium thiosulfate (15 mL) and extracted with ethyl acetate (15 mL. Times.3). The organic phases are combined and dried, filtered and concentrated to obtain a crude product. Purification by column chromatography on silica gel (petroleum ether: ethyl acetate=3:1) gave a white solid (80.0 mg). LC-MS: [ M ] t Bu+H] + :328.2,330.2。
Synthesis of 5-bromo-7- (2-methoxyethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (2):
5-bromo-7- (2-hydroxyethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (70.0 mg,0.18 mmol) was dissolved in tetrahydrofuran (8 mL), 1M lithium bis trimethylsilylamide (0.2 mL,0.20 mmol) was added dropwise at-70℃and stirred at-70℃for half an hour. Methyl iodide (38.8 mg,0.27 mmol) was added dropwise to the reaction system,the reaction was warmed to room temperature and stirred for 0.5 hours. The reaction was quenched with saturated aqueous ammonium chloride (10 mL), extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated aqueous sodium chloride (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give a white solid (28.0 mg). LC-MS: [ M+H ]] + :398.0,400.0。
Synthesis of 7- (2-methoxyethyl) -1, 1-dimethyl-3-oxo-5-boronic acid pinacol ester-isoindoline-2-carboxylic acid tert-butyl ester (3):
the compound tert-butyl 5-bromo-7- (2-methoxyethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylate (28.0 mg,0.07 mmol), pinacol biborate (19.6 mg,0.077 mmol), potassium acetate (17.3 mg,0.18 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (5.1 mg, 0.0071 mmol) was suspended in anhydrous dioxane (3 mL) and reacted under nitrogen at 100deg.C for 5 hours. The reaction solution was diluted with water (5 mL), extracted with ethyl acetate (5 mL. Times.3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product (30.0 mg). The crude product was used directly in the next reaction. LC-MS: [ M+H ] ] + :446.2。
Synthesis of 5- (2-chloro-5-fluoropyrimidin-4-yl) -7- (2-methoxyethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (4):
the compound 7- (2-methoxyethyl) -1, 1-dimethyl-3-oxo-5-boronic acid pinacol ester-isoindoline-2-carboxylic acid tert-butyl ester (30.0 mg,0.067 mmol), 2, 4-dichloro-5-fluoropyrimidine (22.5 mg,0.135 mmol), potassium phosphate (42.9 mg,0.20 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (4.9 mg,0.0067 mmol) was suspended in anhydrous dioxane (4 mL) and water (0.8 mL) and reacted for 4 hours at 100deg.C under nitrogen. The crude product obtained by concentrating the reaction solution was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:1) to obtain a brown solid (22.0 mg). LC-MS: [ M+H ]] + :450.3,452.3。
Synthesis of tert-butyl 5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -7- (2-methylethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylate (5):
5- (2-chloro-5-fluoropyrimidin-4-yl) -7- (2-methoxy)Tert-butyl (17.0 mg,0.037 mmol) of ethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylate, 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (common int-16) (12.5 mg,0.056 mmol), cesium carbonate (36.9 mg,0.11 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (4.7 mg,0.0075 mmol) and palladium acetate (0.8 mg,0.0035 mmol) were dissolved in toluene (1 mL) and reacted at 100℃for 5 hours under nitrogen protection. The crude product obtained by concentration of the reaction solution was purified by preparative TLC (dichloromethane: methanol=10:1) to give a white solid (19.0 mg). LC-MS: [ M+H ] ] + :635.4。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4- (2-methylethyl) -3, 3-dimethylisoindol-1-one (DP-01-210-2):
5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -7- (2-methylethyl) -1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (17.0 mg,0.027 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was reacted at room temperature for 1 hour. The reaction solution was concentrated in vacuo. Dichloromethane (1 mL) and solid sodium carbonate (3 mg) were added to the crude product, and stirred for 1 hour. The organic phase was purified by preparative TLC (dichloromethane: methanol=10:1) to give a white solid (11.1 mg). 1 H NMR(400MHz,DMSO-d6)δ8.65(d,J＝3.2Hz,1H),8.24(s,1H),8.10(s,2H),7.84(d,J＝2.8Hz,1H),6.93(d,J＝8.8Hz,1H),6.61(dd,J＝8.8,2.8Hz,1H),4.94(s,1H),3.80(s,3H),3.71(t,J＝6.8Hz,2H),3.05(t,J＝6.8Hz,6H),2.96(s,3H),2.49–2.38(m,4H),2.23(s,3H),1.55(s,6H)；LC-MS:[M+H] + :535.2。
EXAMPLE 54 Synthesis of Compound DP-01-205
Synthesis of methyl (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) mesylate (1):
methanesulfonyl chloride (139.7 mg,1.22 mmol) was added dropwise under nitrogen to anhydrous dichloro of triethylamine (164.5 mg,1.63 mmol) and (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (common int-14) (300.0 mg,0.81 mmol)In a solution of methane (5 mL) and stirred at room temperature for 1 hour. After completion of the reaction, a saturated aqueous sodium hydrogencarbonate solution (10 mL) was added to the reaction mixture, and the mixture was extracted with methylene chloride (20 mL. Times.2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (300.0 mg). LC-MS: [ M+H ] ] + :446.9,448.9。
Synthesis of 2- (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (2):
to a solution of methyl (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanesulfonate (300.0 mg,0.67 mmol) in N, N-dimethylformamide (5 mL) was added sodium cyanide (92.6 mg,1.90 mmol) and potassium iodide (21.6 mg,0.13 mmol) in this order, and the reaction was stirred at 100℃for 16 hours. After the reaction was completed, the reaction solution was diluted with saturated aqueous sodium hydrogencarbonate (20 mL) and extracted with ethyl acetate (20 mL. Times.2). The organic phases were combined, washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (180.0 mg). LC-MS: [ M+H ]] + :377.9,379.9。
Synthesis of 2- (4- (3- (diphenylmethylene) amino) -4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (3):
to a solution of 2- (4- (3-bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (180.0 mg,0.48 mmol) in 1, 4-dioxane (3 mL) was added benzophenone imine (258.8 mg,1.43 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (59.3 mg,0.095 mmol), tris (dibenzylideneacetone) dipalladium (87.2 mg,0.095 mmol) and cesium carbonate (465.2 mg,1.43 mmol) in this order under nitrogen, and heated to 100℃with stirring for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (200.0 mg). LC-MS: [ M+H ] ] + :479.2。
Synthesis of 2- (4- (3-amino-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (4):
to 2- (4- (3- (diphenylmethylene) amino) -4- (trifluoromethoxy) phenyl) -1-methylpiperazine-2-To a solution of acetonitrile (200.0 mg,0.42 mmol) in 1, 4-dioxane (3.0 mL) was added 4M dioxane hydrochloride solution (3 mL,12 mmol), and the reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give a pale yellow oil (105.0 mg). The crude product was used directly in the next reaction. LC-MS: [ M+H ]] + :315.3。
Synthesis of 2- (4- (3- ((5-fluoro-4- (3 ' -oxospiro [ cyclopropane-1, 1' -isoindoline ] -5' -yl) pyrimidin-2-yl) amino) -4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (DP-01-205):
to a solution of 2- (4- (3-amino-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) acetonitrile (110.0 mg,0.35 mmol) in 1, 4-dioxane (10 mL) was added successively 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindoline]3' -Ketone (common int-10) (153.5 mg,0.53 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (43.6 mg,0.07 mmol), tris (dibenzylideneacetone) dipalladium (32.1 mg,0.035 mmol) and cesium carbonate (342.1 mg,1.05 mmol) were heated to 100℃and stirred for 4 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by prep. HPLC to give a yellow solid (5.1 mg). 1 H NMR(400MHz,DMSO-d6)δ9.15(s,1H),8.88(s,1H),8.63(d,J＝3.5Hz,1H),8.29(s,1H),8.21(d,J＝8.3Hz,1H),7.51–7.49(m,2H),7.24(d,J＝9.0Hz,1H),6.77(dd,J＝9.2,2.9Hz,1H),3.60(d,J＝11.2Hz,1H),3.47(d,J＝13.1Hz,1H),2.96–2.81(m,3H),2.75–2.63(m,2H),2.46(s,1H),2.34(t,J＝9.7Hz,1H),2.27(s,3H),1.54(s,4H)；LC-MS:[M+H] + :568.2。
EXAMPLE 55 Synthesis of Compounds DP-01-208
Synthesis of 5' - (2-chloro-5-fluoro-6-methylpyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (1):
spiro [ cyclopropane-1, 1 '-isoindoline ] to 5' -boronic acid pinacol ester at room temperature]To a solution of 3 '-ketone (common int-9) (300.0 mg,1.05 mmol) in dioxane (5 mL) and water (1 mL) was added 2, 4-dichloro-5-fluoro-6-methylpyrimidine (228.5 mg,1.26 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (79.8 mg,0.11 mmol) and potassium phosphate (669.9 mg,3.16 mmol). The nitrogen was replaced three times and the resulting mixture was stirred at 90℃for 4 hours. The reaction mixture was cooled, quenched with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, washed with saturated brine (10 mL. Times.3), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (petroleum ether: ethyl acetate=5:1) afforded a white solid (200.0 mg). LC-MS: [ M+H ]] + :304.2。
Synthesis of 5' - (5-fluoro-6-methyl-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-208):
at room temperature, 5'- (2-chloro-5-fluoro-6-methylpyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]To a solution of 3' -ketone (78.9 mg,0.26 mmol) and 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (common int-1) (88.1 mg,0.32 mmol) in 1, 4-dioxane (1 mL) was added cesium carbonate (257.5 mg,0.79 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (32.8 mg,0.053 mmol) and tris (dibenzylideneacetone) dipalladium (24.1 mg,0.026 mmol). After three nitrogen substitutions, the mixture was reacted at 90℃for 6 hours. After the reaction was complete, the excess solvent was removed in vacuo, diluted with water (10 mL) and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Purification by basic preparation gave a white solid (11.8 mg). 1 H NMR(400MHz,DMSO-d6)δ8.87–8.85(m,2H),8.25(s,1H),8.17(d,J＝8.2Hz,1H),7.64(s,1H),7.46(d,J＝8.3Hz,1H),7.19(d,J＝8.0Hz,1H),6.79–6.63(m,1H),3.15(s,4H),2.45–2.42(m,7H),2.21(s,3H),1.53(s,4H)；LC-MS:[M+H] + :543.4。
EXAMPLE 56 Synthesis of Compounds DP-01-219, DP-01-219A, DP-01-219B
Synthesis of (4- (3- (diphenylmethylene) amino) -4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (1):
direction (4- (3)To a solution of 1, 4-dioxan (3 mL) in bromo-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (common int-14) (210.0 mg,0.57 mmol) was added, in order, benzophenone imine (206.2 mg,1.14 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (68.5 mg,0.11 mmol), tris (dibenzylideneacetone) dipalladium (100.7 mg,0.11 mmol) and cesium carbonate (556.0 mg,1.71 mmol), and the mixture was heated to 100℃under nitrogen atmosphere and stirred for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (160.0 mg). LC-MS: [ M+H ]] + :470.1。
Synthesis of (4- (3-amino-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (2):
(4- (3- (diphenylmethylene) amino) -4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (160.0 mg,0.34 mmol) was dissolved in 1, 4-dioxane (3 mL), a 4M dioxane hydrochloride solution (3 mL,12 mmol) was added, and the reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to give a pale yellow oil (50.0 mg). The crude product was used directly in the next reaction. LC-MS: [ M+H ] ] + :306.3。
Synthesis of 5' - (5-fluoro-2- ((5- (3- (hydroxymethyl) -4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-219):
to a solution of (4- (3-amino-4- (trifluoromethoxy) phenyl) -1-methylpiperazin-2-yl) methanol (50.0 mg,0.16 mmol) in 1, 4-dioxane (10 mL) was added successively 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindoline]3' -Ketone (common int-10) (57.0 mg,0.20 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (20.4 mg,0.033 mmol), tris (dibenzylideneacetone) dipalladium (15.0 mg,0.016 mmol) and cesium carbonate (160.3 mg,0.49 mmol) were heated to 100℃and stirred for 4 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by prep. to give a yellow solid (4.2 mg). 1 H NMR(400MHz,DMSO-d6)δ9.11(s,1H),8.88(s,1H),8.62(d,J＝3.4Hz,1H),8.28(s,1H),8.20(d,J＝8.4Hz,1H),7.52–7.44(m,2H),7.22(d,J＝9.1Hz,1H),6.75(d,J＝9.2Hz,1H),4.57(s,1H),3.64(d,J＝10.7Hz,2H),3.51(d,J＝10.8Hz,1H),3.39(s,1H),2.81(s,2H),2.61–2.54(m,1H),2.35–2.27(m,1H),2.25(s,3H),2.13(s,1H),1.54(s,4H)；LC-MS:[M+H] + :559.3。
The racemate DP-01-219 was purified by SFC separation (column: CHIRALPAK C-IG 5 μm 4.6 x 100mm I.D.,5 μm; mobile phase: A phase: CO) 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: etOH (0.05% DEA v/v); gradient: b%:40.0%,8 min) to give white solids DP-01-219A and white solids DP-01-219B.
DP-01-219A：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, 5 μm mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: etOH (0.05% DEA v/v); gradient: b%:40% with retention time 2.727min; ee% = 100.0%.
1 H NMR(400MHz,DMSO-d6)δ9.10(s,1H),8.87(s,1H),8.62(d,J＝3.5Hz,1H),8.27(s,1H),8.20(d,J＝8.1Hz,1H),7.46(dd,J＝11.1,5.5Hz,2H),7.21(d,J＝8.6Hz,1H),6.74(dd,J＝9.1,2.8Hz,1H),4.56(t,J＝5.4Hz,1H),3.63(d,J＝11.0Hz,2H),3.50(d,J＝11.8Hz,1H),3.36(s,1H),2.80(s,2H),2.60–2.53(m,1H),2.26–2.22(m,4H),2.12(s,1H),1.53(s,4H)；LC-MS:[M+H] + :559.2。
DP-01-219B：
Chiral analysis conditions: chromatographic column: CHIRALPAK C-IG 5 μm 4.6 x 100mm, 5 μm mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: etOH (0.05% DEA v/v); gradient: b%:40% with a retention time of 3.532min; ee% = 97.6%.
1 H NMR(400MHz,DMSO-d6)δ9.10(s,1H),8.87(s,1H),8.62(d,J＝3.5Hz,1H),8.27(s,1H),8.20(d,J＝8.4Hz,1H),7.46(dd,J＝11.3,5.4Hz,2H),7.21(d,J＝8.6Hz,1H),6.74(dd,J＝9.1,2.8Hz,1H),4.56(t,J＝5.5Hz,1H),3.63(d,J＝10.9Hz,2H),3.50(d,J＝11.2Hz,1H),3.36(s,1H),2.80(s,2H),2.60–2.53(m,1H),2.31–2.22(m,4H),2.12(s,1H),1.53(s,4H)；LC-MS:[M+H] + :559.2。
EXAMPLE 57 Synthesis of Compounds DP-01-229
(E) -synthesis of tert-butyl 6-bromo-4- (2-ethoxyvinyl) -3-methyl-1-isoindoline-2-carboxylate (1):
the compound tert-butyl 6-bromo-4-iodo-3-methyl-1-oxoisoindole-2-carboxylate (common int-15) (700.0 mg,1.55 mmol), (E) -1-ethoxyvinyl-2-boronic acid pinacol ester (337.3 mg,1.56 mmol), potassium phosphate (657.3 mg,3.10 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (113.3 mg,0.156 mmol) was placed in dioxane (10 mL) and water (1 mL) and reacted at 100℃for 3 hours under nitrogen. The reaction solution was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate=100:1-50:1) to give a white oily compound (350.0 mg). 1 H NMR(400MHz,DMSO-d6)δ7.95(s,1H),7.60(s,1H),7.48(d,J＝12.8Hz,1H),5.89(d,J＝12.8Hz,1H),5.23(d,J＝6.4Hz,1H),4.04–3.95(m,2H),1.53(d,J＝1.2Hz,9H),1.50–1.47(m,3H),1.32–1.26(m,3H).
Synthesis of 2- (6-bromo-3-methyl-1-oxoisoindol-4-yl) acetaldehyde (2):
the compound (E) -6-bromo-4- (2-ethoxyvinyl) -3-methyl-1-isoindoline-2-carboxylic acid tert-butyl ester (310.0 mg,0.78 mmol) was dissolved in formic acid (1.5 mL), heated to 50℃and stirred for 1 hour. The reaction mixture was diluted with water (10 mL), made weakly basic with saturated sodium bicarbonate solution, and extracted with ethyl acetate (3X 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give a white solid (120.0 mg). The crude product was used directly in the next reaction. LC-MS: [ M+H ] + ]:268.0,270.0。
Synthesis of 6-bromo-4- (2-hydroxyethyl) -3-methylisoindol-1-one (3):
the compound 2- (6-bromo-3-methyl-1-oxoisoindol-4-yl) acetaldehyde (115.0 mg,0.43 mmol) was dissolved in ethanol (1 mL) and sodium borohydride (24.5 mg,0.65 mmol) was added at 0deg.C and reacted at room temperature for 1.5 hours. The reaction mixture was diluted with 5mL of water and extracted with ethyl acetate (3X 10 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give a crude white solid (60.0 mg). LC-MS: [ M+H ] + ]:269.9,271.9。
Synthesis of 4- (2-hydroxyethyl) -3-methyl-6-boronic acid pinacol ester isoindol-1-one (4):
will be converted intoCompound 6-bromo-4- (2-hydroxyethyl) -3-methylisoindol-1-one (55.0 mg,0.20 mmol), pinacol biborate (60.9 mg,0.24 mmol), potassium acetate (59.9 mg,0.61 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (14.8 mg,0.02 mmol) was placed in anhydrous dioxane (1 mL). The reaction was stirred at 100℃for 3 hours under nitrogen. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane: methanol=50:1 to 20:1) to give a white solid compound (37.0 mg). LC-MS: [ M+H ]] + :318.1。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3-methylisoindol-1-one (5):
the compound 4- (2-hydroxyethyl) -3-methyl-6-boronic acid pinacol ester isoindol-1-one (30.1 mg,0.095 mmol), 2, 4-dichloro-5-fluoropyrimidine (18.4 mg,0.11 mmol), potassium phosphate (41.0 mg,0.19 mmol) and [1,1' -bis (diphenylphosphine) ferrocene ]Palladium dichloride (6.9 mg,0.0095 mmol) was placed in dioxane (1 mL) and water (0.2 mL) and the reaction stirred at 100deg.C under nitrogen for 3 hours. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane: methanol=30:1 to 10:1) to give a white solid (20.0 mg). LC-MS: [ M+H ]] + :322.0。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4- (2-hydroxyethyl) -3-methylisoindol-1-one (DP-01-229):
the compound 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3-methylisoindol-1-one (16.1 mg,0.05 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (common int-16) (12.9 mg,0.06 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (6.6 mg,0.01 mmol), palladium acetate (2.2 mg,0.01 mmol) and cesium carbonate (32.5 mg,0.10 mmol) were placed in anhydrous dioxane (0.5 mL) and reacted under nitrogen protection at 90℃for 3 hours with stirring. The reaction was diluted with 5mL of water and extracted with ethyl acetate (3X 5 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give a yellow solid (2.1 mg) which was isolated and purified. 1 H NMR(400MHz,DMSO-d6)δ8.78(s,1H),8.65(d,J＝3.6Hz,1H),8.21(s,1H),8.09(s,2H),7.87(d,J＝2.8Hz,1H),6.93(d,J＝8.8Hz,1H),6.61(dd,J＝8.8,2.8Hz,1H),4.93–4.78(m,2H),3.81(s,3H),3.76–3.66(m,2H),3.06–3.01(m,4H),2.99–2.89(m,2H),2.47–2.42(m,4H),2.20(s,3H),1.44(d,J＝6.6Hz,3H)；LC-MS:[M+H] + :507.2。
EXAMPLE 58 Synthesis of Compounds DP-01-146
Synthesis of tert-butyl 4- (3-amino-4- (trifluoromethoxy) phenyl) piperazine-1-carboxylate (2):
The compound 5-bromo-2- (trifluoromethoxy) aniline (2.0 g,7.81 mmol), tris (dibenzylideneacetone) dipalladium (0.7 g,0.76 mmol), 2-dicyclohexylphosphino-2' - (N, N-dimethylamine) -biphenyl (0.6 g,1.52 mmol) was dissolved in tetrahydrofuran (20 mL), replaced three times with nitrogen, 1M lithium bis (trimethylsilyl) amide (18.8 mL,18.80 mmol) was added dropwise, piperazine-1-carboxylate (2.3 g,12.35 mmol) was added, the temperature was raised to 70 ℃ and stirred for 4 hours, LCMS detection was complete, water (50 mL) was added to the reaction solution, extraction was performed with ethyl acetate (50 ml×3), the combined organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, rotary distilled to give crude product, and red solid (998.0 mg) was obtained by reverse column chromatography (acetonitrile: water=5-60%). LC-MS: [ M+H ]] + :362.2。
Synthesis of tert-butyl 4- (3- ((5-fluoro-4- (3 ' -oxospiro [ cyclopropane-1, 1' -isoindoline ] -5' -yl) pyrimidin-2-yl) amino) -4- (trifluoromethoxy) phenyl) piperazine-1-carboxylate (3):
tert-butyl 4- (3-amino-4- (trifluoromethoxy) phenyl) piperazine-1-carboxylate (74.9 mg,0.21 mmol) and 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (common int-10) (50.0 mg,0.17 mmol) was dissolved in 1, 4-dioxane, and cesium carbonate (168.7 mg,0.52 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (24.9 mg,0.04 mmol) and tris (dibenzylideneacetone) dipalladium (18.3 mg,0.02 mmol) were added to the reaction flask and nitrogen sparged for 1 minute. The reaction mixture was reacted at 100℃for 16 hours. LCMS detected complete reaction. The reaction solution was poured into water (10 mL) at room temperature, and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give a yellow solid (30.0 mg). LC-MS: [ M+H ] ] + :615.2。
Synthesis of 5' - (5-fluoro-2- ((5- (piperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-146):
4- (3- ((5-fluoro-4- (3 '-oxospiro [ cyclopropane-1, 1' -isoindoline)]-5' -yl) pyrimidin-2-yl) amino) -4- (trifluoromethoxy) phenyl) piperazine-1-carboxylic acid tert-butyl ester (30.0 mg,0.05 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.5 mL) was added to the reaction solution and stirred at room temperature for 10 hours. LCMS detected complete reaction. The reaction solution was concentrated under reduced pressure, and a yellow solid (6.1 mg) was obtained by separation and purification. 1 H NMR(400MHz,DMSO)δ9.05(s,1H),8.86(s,1H),8.63(s,1H),8.29(s,1H),8.19(d,J＝9.2Hz,1H),7.54(s,1H),7.49–7.46(m,1H),7.20–7.18(m,1H),6.73–7.71(m,1H),3.05(s,4H),2.82(s,4H),1.53(s,4H),1.23(s,1H)；LC-MS:[M+H] + :515.2。
EXAMPLE 59 Synthesis of Compound DP-01-200-2
Synthesis of 5-bromo-7-formyl-1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (1):
5-bromo-1, 1-dimethyl-3-oxo-7-vinylisoindole-2-carboxylic acid tert-butyl ester (common int-17) (200.0 mg,0.55 mmol) was dissolved in isopropanol (5 mL), sodium periodate (233.6 mg,1.08 mmol) was added, followed by potassium osmium sulfate dihydrate (8.50 mg,0.023 mmol). The reaction solution was stirred at room temperature for 4 hours. The reaction solution was poured into ice water (10 mL), extracted with ethyl acetate (10 mL. Times.3), and the organic phase was dried and concentrated to give a white solid (190.0 mg). LC-MS [ M-t-Bu+H ]] + :312.1,314.1。
Synthesis of 5-bromo-7- (hydroxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (2):
5-bromo-7-formyl-1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (170.0 mg,0.46 mmol) was dissolved in ethanol (5 mL), and sodium borohydride (26.2 mg,0.69 mmol) was added under ice-bath and reacted at room temperature for 1 hour. The reaction mixture was dried by spinning, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL. Times.3), and the organic phase was extractedDrying and spin-drying gave a white solid (90.0 mg). LC-MS [ M-t-Bu+H ]] + :314.2,316.2。
Synthesis of 5-bromo-7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (3):
5-bromo-7- (hydroxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (70.0 mg,0.19 mmol) was dissolved in tetrahydrofuran (5 mL), 1M lithium bis trimethylsilylamide (0.3 mL,0.30 mmol) was added dropwise at-70℃and stirred for half an hour at-70 ℃. Methyl iodide (53.7 mg,0.38 mmol) was added dropwise to the reaction system, the reaction was warmed to room temperature and stirred for 0.5 hours, quenched with saturated aqueous ammonium chloride (10 mL), extracted with ethyl acetate (10 mL. Times.3), the organic phase was washed with saturated aqueous sodium chloride (20 mL), dried, and spun-dried, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give a white solid (30.0 mg), LC-MS: [ M+H] + :384.2,386.2。
Synthesis of 7- (methoxymethyl) -1, 1-dimethyl-3-oxo-5-boronic acid pinacol ester isoindole-2-carboxylic acid tert-butyl ester (4):
The compound tert-butyl 5-bromo-7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylate (30.0 mg,0.078 mmol), pinacol biborate (21.8 mg,0.086 mmol), potassium acetate (19.2 mg,0.20 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (5.71 mg,0.0079 mmol) was suspended in anhydrous dioxane (3 mL), reacted at 95℃for 18 hours under nitrogen protection, water (5 mL) was added to the system, followed by extraction with ethyl acetate (5 mL. Times.3), the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product (30.0 mg). LC-MS [ M-t-Bu+H ]] + :432.3。
Synthesis of 5- (2-chloro-5-fluoropyrimidin-4-yl) -7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (5):
the compound 7- (methoxymethyl) -1, 1-dimethyl-3-oxo-5-boronic acid pinacol ester isoindole-2-carboxylic acid tert-butyl ester (30.0 mg,0.069 mmol), 2, 4-dichloro-5-fluoropyrimidine (25.0 mg,0.15 mmol), potassium phosphate (44.3 mg,0.21 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (17.0 mg,0.023 mmol) was suspended in anhydrous dioxane (4 mL) and water (0.8 mL) and the strips were incubated at 100deg.C under nitrogenThe reaction was carried out for 4 hours under the column. The reaction solution was dried by spin-drying, and the crude product was subjected to silica gel column chromatography (dichloromethane: methanol=10:1) to give a brown solid (30.0 mg). LC-MS: [ M+H ] ] + :436.3。
Synthesis of 5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (6):
tert-butyl 5- (2-chloro-5-fluoropyrimidin-4-yl) -7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylate (20.0 mg,0.046 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (common int-16) (15.2 mg,0.068 mmol), cesium carbonate (44.8 mg,0.14 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (5.7 mg,0.009 mmol) and palladium acetate (1.6 mg, 0.0079 mmol) were dissolved in 1, 4-dioxane (1 mL), reacted at 100 ℃ for 5 hours under nitrogen protection, and the crude product was dried by spin-drying on a silica gel plate (dichloromethane: methanol=10:1) to give a white solid (9.0 mg). LC-MS: [ M+H ]] + :621.2。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4- (methoxymethyl) -3, 3-dimethylisoindol-1-one (DP-01-200-2):
5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -7- (methoxymethyl) -1, 1-dimethyl-3-oxoisoindole-2-carboxylic acid tert-butyl ester (9.0 mg,0.015 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.5 mL,6.53 mmol) was added, and the reaction mixture was dried by spinning at room temperature for 1 hour. Dichloromethane (1 mL) and sodium carbonate (3 mg) were added and stirred for 1 hour, and the organic phase was prepared as a white solid (5.3 mg) via a silica gel preparation plate (dichloromethane: methanol=10:1). 1 H NMR(400MHz,DMSO)δ8.66(d,J＝3.2Hz,1H),8.37(s,1H),8.23(s,1H),8.18(s,1H),7.87(d,J＝2.8Hz,1H),6.93(d,J＝8.8Hz,1H),6.61(dd,J＝8.8,2.8Hz,1H),5.61(s,1H),4.83(s,2H),3.81(s,3H),3.05(s,4H),2.96(s,3H),2.50(s,4H),2.25(s,3H),1.53(s,6H)；LC-MS:[M+H] + :521.3。
EXAMPLE 60 Synthesis of Compound DP-01-223
Synthesis of 6-bromo-3, 3-dimethyl-4- (S-methylsulfonylimino) isoindolin-1-one (1):
the compound 6-bromo-3, 3-dimethyl-4- (methylthio) isoindolin-1-one (200.0 mg,0.70 mmol), iodobenzene diacetic acid (563.0 mg,1.75 mmol), ammonium carbamate (109.0 mg,1.40 mmol) was dissolved in methanol (2.0 mL). The reaction solution was stirred at room temperature for 12 hours. After the reaction was complete, the excess solvent was removed in vacuo. The crude product was purified by column chromatography (dichloromethane: methanol=20:1) to give a yellow oily liquid (107.0 mg). LC-MS: [ M+H ]] + :317.0,319.0。
Synthesis of (1, 1-dimethyl-7- (S-methylsulfonylimino) -3-oxoisoindolin-5-yl) boronic acid (2):
the compound 6-bromo-3, 3-dimethyl-4- (S-methylsulfonylimino) isoindolin-1-one (107.0 mg,0.34 mmol), [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (24.9 mg,0.034 mmol), pinacol biborate (111.3 mg,0.44 mmol), potassium acetate (99.2 mg,1.01 mmol) were placed in 1, 4-dioxane (2.0 mL). The reaction system was stirred at 80℃for 2 hours under nitrogen protection. After the reaction was completed, the reaction solution was directly concentrated to give a black solid (95.9 mg). LC-MS: [ M+H ]] + :283.0。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethyl-4- (S-methylsulfonylimino) isoindolin-1-one (3):
(1, 1-dimethyl-7- (S-methylsulfonylimino) -3-oxoisoindolin-5-yl) boronic acid (95.9 mg,0.34 mmol), 2, 4-dichloro-5-fluoropyrimidine (67.5 mg,0.40 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (24.7 mg,0.034 mmol), potassium phosphate (214.6 mg,1.01 mmol) were placed in a mixed solution of 1, 4-dioxane and water (2.2 mL,1, 4-dioxane: water=10:1, v/v). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (dichloromethane: methanol=10:1) afforded a tan solid (98.5 mg). LC-MS: [ M+H ]] + :369.0。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethyl-4 (S-methylsulfonylimino) isoindolin-1-one (DP-01-223):
2-methoxy-5- (4-methylpiperazin-1-yl) aniline (75.5 mg,0.34 mmol), 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethyl-4- (S-methylsulfonylamino) isoindolin-1-one (98.5 mg,0.27 mmol), tris (dibenzylideneacetone) dipalladium (28.4 mg,0.031 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (38.6 mg,0.062 mmol), cesium carbonate (202.0 mg,0.62 mmol) were placed in 1, 4-dioxane (1.0 mL). The reaction system was stirred at 100℃for 2 hours under nitrogen protection. After completion of the reaction, water (10.0 mL) was added thereto, and the mixture was quenched and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The yellow solid (4.3 mg) was obtained by preparative high performance liquid chromatography. 1 H NMR(400MHz,CDCl 3 )δ9.12(s,1H),8.80(s,1H),8.46(s,1H),8.32(s,1H),7.90(s,1H),6.84(d,J＝8.4Hz,1H),6.59(d,J＝8.0Hz,1H),6.40(s,1H),3.91(s,3H),3.21(s,4H),3.16(s,3H),2.79(s,4H),2.47(s,3H),1.93–1.90(m,6H)；LC-MS:[M+H] + :554.2。
EXAMPLE 61 Synthesis of Compound DP-01-225
Synthesis of 6-bromo-3, 3-dimethyl-4- (methylsulfonyl) isoindolin-1-one (1):
the compound 6-bromo-3, 3-dimethyl-4- (methylthio) isoindolin-1-one (500.0 mg,1.75 mmol) and potassium hydrogen peroxysulfate complex salt (3.22 g,9.30 mmol) were placed in a mixed solution of tetrahydrofuran and water (20 mL, tetrahydrofuran: water=1:1). The reaction system was stirred at room temperature for 12 hours. After completion of the reaction, the reaction was quenched with water (50.0 mL) and extracted with ethyl acetate (50 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (dichloromethane: methanol=20:1) to give a yellow solid (200.0 mg). LC-MS: [ M+H ]] + :318.0,319.9。
Synthesis of (1, 1-dimethyl-7- (methylsulfonyl) -3-oxoisoindolin-5-yl) boronic acid (2):
the compound 6-bromo-3, 3-dimethyl-4- (methylsulfonyl) isoindolin-1-one (24.0 mg,0.075 mmol) was reacted with [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (5.5 mg,0.0075 mmol), pinacol biborate (24.9 mg,0.098 mmol), potassium acetate (22.2 mg,0.23 mmol) were placed in 1, 4-dioxane (1.0 mL). The reaction system was stirred at 80℃for 2 hours under nitrogen protection. After the reaction was completed, the reaction solution was directly concentrated to obtain a crude black solid (46.3 mg) which was directly used for the next reaction. LC-MS: [ M+H ] ] + :284.1。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethyl-4- (methylsulfonyl) isoindolin-1-one (3):
(1, 1-dimethyl-7- (methylsulfonyl) -3-oxoisoindolin-5-yl) boronic acid (22.6 mg,0.08 mmol), 2, 4-dichloro-5-fluoropyrimidine (15.1 mg,0.09 mmol) and [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (5.5 mg,0.0075 mmol), potassium phosphate (48.0 mg,0.23 mmol) were placed in a mixed solution of 1, 4-dioxane and water (1.1 mL,1, 4-dioxane: water=10:1). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After the reaction was completed, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (dichloromethane: methanol=10:1) afforded a yellow solid (14.8 mg). LC-MS: [ M+H ]] + :370.0,372.0。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethyl-4- (methylsulfonyl) isoindolin-1-one (DP-01-225):
6- (2-chloro-5-fluoropyrimidin-4-yl) -3, 3-dimethyl-4- (methylsulfonyl) isoindolin-1-one (15.0 mg,0.04 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (9.90 mg,0.04 mmol), tris (dibenzylideneacetone) dipalladium (7.4 mg,0.008 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (10.1 mg,0.016 mmol), cesium carbonate (52.8 mg,0.16 mmol) were placed in 1, 4-dioxane (1.0 mL). The reaction system was stirred at 100℃for 2 hours under nitrogen protection. To be reversed The reaction was quenched by the addition of water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The mixture was purified by preparative separation to give a yellow solid (7.0 mg). 1 H NMR(400MHz,MeOD)δ8.91(s,1H),8.73(s,1H),8.56(d,J＝3.2Hz,1H),8.17(d,J＝2.4Hz,1H),6.93(d,J＝8.8Hz,1H),6.64(dd,J＝8.8,2.4Hz,1H),3.89(s,3H),3.23(s,3H),3.16–3.09(m,4H),2.66–2.56(m,4H),2.32(s,3H),1.86(s,6H)；LC-MS:[M+H] + :555.2。
EXAMPLE 62 Synthesis of Compounds DP-01-239
Synthesis of 1- (4-bromo-2-nitrophenoxy) -2-methylpropan-2-ol (2):
the compound 4-bromo-1-fluoro-2-nitrobenzene (500.0 mg,2.27 mmol), 2-methylpropane-1, 2-diol (266.3 mg,2.95 mmol) and cesium carbonate (1.5 g,4.60 mmol) were placed in N, N-dimethylformamide (10 mL), nitrogen was replaced, and the temperature was raised to 80℃and stirred for 6 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL. Times.3). The combined organic phases were washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography on silica gel (ethyl acetate: petroleum ether=0-25%) to give a yellow oil (461.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.10(d,J＝2.4Hz,1H),7.80(dd,J＝9.0,2.4Hz,1H),7.34(d,J＝9.0Hz,1H),4.70(s,1H),3.89(s,2H),1.19(s,6H)。
Synthesis of 2-methyl-1- (4- (4-methylpiperazin-1-yl) -2-nitrophenoxy) propan-2-ol (3):
the compound 1- (4-bromo-2-nitrophenoxy) -2-methylpropan-2-ol (200.0 mg,0.69 mmol), 1-methylpiperazine (82.9 mg,0.83 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (56.6 mg,0.14 mmol), cesium carbonate (449.3 mg,1.38 mmol) and tris (dibenzylideneacetone) dipalladium (63.1 mg,0.069 mmol) were placed in 1, 4-dioxane (5 mL), purged with nitrogen three times, and heated to 90 ℃ and stirred for 6 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL. Times.3). Combining organic phases And brine (40 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. Purification by silica gel column chromatography (methanol: dichloromethane=0-5%) gave a yellow oil (160.0 mg). LC-MS: [ M+H ]] + :310.2。
Synthesis of 1- (2-amino-4- (4-methylpiperazin-1-yl) phenoxy) -2-methylpropan-2-ol (4):
the compound 2-methyl-1- (4- (4-methylpiperazin-1-yl) -2-nitrophenoxy) propan-2-ol (150.0 mg,0.48 mmol) and palladium on carbon (50.0 mg,10% w/w) were suspended in methanol (4 mL), and after three hydrogen substitutions, the reaction was stirred at room temperature for 4 hours. After completion of the reaction by LCMS, the reaction mixture was filtered through celite, the filter cake was washed 3 times with ethyl acetate, and the filtrate was dried to give a yellow solid (128.0 mg). LC-MS: [ M+H ]] + :280.4.5'- (5-fluoro-2- ((2- (2-hydroxy-2-methylpropyloxy) -5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-synthesis of 3' -ketone (DP-01-239):
the compound 1- (2-amino-4- (4-methylpiperazin-1-yl) phenoxy) -2-methylpropan-2-ol (128.0 mg,0.46 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindoline]3' -Ketone (159.3 mg,0.55 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (57.0 mg,0.092 mmol), cesium carbonate (298.5 mg,0.92 mmol) and tris (dibenzylideneacetone) dipalladium (41.9 mg,0.046 mmol) were placed in 1, 4-dioxane (5 mL), nitrogen was replaced, and the mixture was stirred at 100℃for 6 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. Purification by prep. gave a yellow solid (41.1 mg). 1 H NMR(400MHz,CDCl 3 )δ8.57(s,1H),8.37–8.27(m,3H),7.95(s,1H),7.66(s,1H),7.25(d,J＝8.4Hz,1H),7.14(d,J＝8.0Hz,1H),6.85(d,J＝8.4Hz,1H),6.57–6.47(m,1H),3.90–3.79(m,2H),3.19(s,4H),2.60(s,4H),2.35(s,3H),1.70–1.63(m,2H),1.55–1.46(m,2H),1.39(s,6H)；LC-MS:[M+H] + :533.8。
EXAMPLE 63 Synthesis of Compounds DP-01-242
Synthesis of 1-methyl-4- (4-methyl-3-nitrophenyl) piperazine (2):
the compound 4-bromo-1-methyl-2-nitrobenzene (300.0 mg,1.39 mmol), 1-methylpiperazine (166.9 mg,1.67 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxy-biphenyl (114.0 mg,0.28 mmol), cesium carbonate (905.0 mg,2.78 mmol) and tris (dibenzylideneacetone) dipalladium (127.2 mg,0.14 mmol) were placed in 1, 4-dioxane (6 mL) under nitrogen, and stirred for 6 hours at 90 ℃. LCMS detected the starting material had reacted. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (40 mL. Times.3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (methanol: dichloromethane=0-5%) to give a yellow oil (216.0 mg). LC-MS: [ M+H ]] + :236.1。
Synthesis of 2-methyl-5- (4-methylpiperazin-1-yl) aniline (3):
the compound 1-methyl-4- (4-methyl-3-nitrophenyl) piperazine (200.0 mg,0.85 mmol) and palladium on carbon (80.0 mg,10% w/w) were suspended in methanol (4 mL), and after three hydrogen substitutions, the reaction was stirred at room temperature for 4 hours. After completion of the reaction by LCMS, the reaction was filtered through celite, the filter cake was washed 3 times with ethyl acetate, and the filtrate was concentrated to a white solid (170.0 mg). LC-MS: [ M+H ] ] + :206.2。
Synthesis of 5' - (5-fluoro-2- ((2-methyl-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-242):
the compound 2-methyl-5- (4-methylpiperazin-1-yl) aniline (120.0 mg,0.58 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (203.2 mg,0.70 mmol), cesium carbonate (380.9 mg,1.17 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (72.8 mg,0.12 mmol) and tris (dibenzylideneacetone) dipalladium (53.5 mg,0.06 mmol) were placed in 1, 4-dioxane (4 mL), nitrogen was replaced, and the mixture was stirred at 100℃for 6 hours. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL. Times.3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. A pale yellow solid (122.9 mg) was obtained. 1 H NMR(400MHz,DMSO-d6)δ8.85–8.82(m,2H),8.54(d,J＝3.6Hz,1H),8.27(s,1H),8.17(d,J＝8.0Hz,1H),7.45(d,J＝8.4Hz,1H),7.16(d,J＝2.0Hz,1H),7.05(d,J＝8.4Hz,1H),6.67(dd,J＝8.0,2.4Hz,1H),3.11–3.03(m,4H),2.46–2.39(m,4H),2.20(s,3H),2.14(s,3H),1.52(s,4H)；LC-MS:[M+H] + :459.6。
EXAMPLE 64 Synthesis of Compound DP-01-252
Synthesis of 2- (4-bromo-2-nitrophenoxy) ethan-1-ol (2):
the compound 4-bromo-1-fluoro-2-nitrobenzene (500.0 mg,2.27 mmol), ethylene glycol (423.4 mg,6.82 mmol), cesium carbonate (2.96 g,9.08 mmol) was placed in N, N-dimethylformamide (10.0 mL). The reaction mixture was stirred at 80℃for 12 hours. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (PE: ea=2:1) gave a yellow oily liquid (360.0 mg).
Synthesis of 2- (4- (4-methylpiperazin-1-yl) -2-nitrophenoxy) ethan-1-ol (3):
2- (4-bromo-2-nitrophenoxy) ethan-1-ol (360.0 mg,1.37 mmol) was dissolved in anhydrous 1, 4-dioxane (6 mL), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (56.4 mg,0.14 mmol), tris (dibenzylideneacetone) dipalladium (63.2 mg,0.069 mmol), cesium carbonate (895.4 mg,2.75 mmol) and N-methylpiperazine (206.5 mg,2.06 mmol) were added sequentially, and the reaction mixture was stirred at 90℃for 12 hours. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, finally dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (DCM: meoh=20:1) afforded a green oily liquid (102.0 mg). LC-MS: [ M+H ]] + :282.2。
Synthesis of 2- (2-amino-4- (4-methylpiperazin-1-yl) phenoxy) ethan-1-ol (4):
the compound 2- (4- (4-methylpiperazin-1-yl) -2-nitrophenoxy) ethan-1-ol (102.0 mg,0.36 mmol) and palladium on carbon (35.0 mg,10% w/w) were placed in methanol (5 mL) and the reaction was stirred at room temperature under hydrogen atmosphere for 2 hours. After the reaction was completed, the reaction solution was filtered through celite, the cake was washed twice with a small amount of methanol, and the filtrate was concentrated to give a brown-yellow solid (98.3 mg). LC-MS: [ M+H ] ] + :252.2。
Synthesis of 5' - (5-fluoro-2- ((2- (2-hydroxyethoxy) -5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-252):
2- (2-amino-4- (4-methylpiperazin-1-yl) phenoxy) ethan-1-ol (98.3 mg,0.39 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (113.3 mg,0.39 mmol), tris (dibenzylideneacetone) dipalladium (32.5 mg,0.035 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (44.2 mg,0.07 mmol), cesium carbonate (231.3 mg,0.71 mmol) were placed in 1, 4-dioxane (2.0 mL). The reaction system was stirred at 100℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Yellow solid (29.4 mg) was isolated by preparative separation. 1 H NMR(400MHz,MeOD)δ8.52(s,1H),8.50(d,J＝3.6Hz,1H),8.40–8.32(m,2H),7.42(d,J＝8.0Hz,1H),6.94(d,J＝8.8Hz,1H),6.61(dd,J＝8.8,2.8Hz,1H),4.17–4.05(m,2H),3.99–3.86(m,2H),3.21–3.09(m,4H),2.71–2.58(m,4H),2.36(s,3H),1.74–1.52(m,4H)；LC-MS:[M+H] + :505.3。
EXAMPLE 65 Synthesis of Compounds DP-01-258
Synthesis of 6- (2- ((2-ethoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (DP-01-258):
6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (50.0 mg,0.15 mmol), 2-ethyl Oxy-5- (4-methylpiperazin-1-yl) aniline (45.6 mg,0.19 mmol), cesium carbonate (145.6 mg,0.45 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (18.6 mg,0.03 mmol) and palladium acetate (3.3 mg,0.015 mmol) were placed in 1, 4-dioxane (1 mL) and reacted at 100℃for 18 hours under nitrogen protection. The reaction solution is cooled to room temperature and then concentrated under reduced pressure to obtain crude products. The crude product was prepared by acidity to give a yellow solid (19.5 mg). 1 H NMR(400MHz,DMSO-d6)δ8.82(s,1H),8.66(d,J＝3.2Hz,1H),8.18–8.15(m,2H),8.08–8.06(m,2H),7.90(d,J＝2.8Hz,1H),6.93(d,J＝8.8Hz,1H),6.59(dd,J＝8.8,2.8Hz,1H),4.06(q,J＝7.2Hz,2H),3.70(t,J＝7.2Hz,2H),3.07–2.99(m,6H),2.48–2.43(m,4H),2.21(s,3H),1.58(s,6H),1.34(t,J＝6.8Hz,3H)；LC-MS:[M+H] + :535.2。
EXAMPLE 66 Synthesis of Compounds DP-01-259
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (DP-01-259):
6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (2-hydroxyethyl) -3, 3-dimethylisoindol-1-one (50.0 mg,0.15 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (49.4 mg,0.22 mmol), cesium carbonate (145.6 mg,0.45 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (18.6 mg,0.03 mmol) and palladium acetate (3.3 mg,0.015 mmol) were suspended in 1, 4-dioxane (1 mL), reacted at 100℃for 18 hours under nitrogen atmosphere, and the reaction mixture was cooled and concentrated. The crude product was prepared by acidity to give a yellow solid (21.5 mg). 1 H NMR(400MHz,DMSO-d6)δ8.82(s,1H),8.66(d,J＝3.2Hz,1H),8.33(s,2H),8.22(s,1H),8.09(s,1H),8.07(s,1H),7.86(d,J＝2.8Hz,1H),6.94(d,J＝9.2Hz,1H),6.62(dd,J＝9.2,2.8Hz,1H),3.81(s,3H),3.70(t,J＝6.8Hz,2H),3.07–2.98(m,6H),2.49–2.40(m,4H),2.21(s,3H),1.58(s,6H)；LC-MS:[M+H] + :521.2。
EXAMPLE 67 Synthesis of Compounds DP-01-261
Synthesis of (1R, 5S) -3- (4-methoxy-3-nitrophenyl) -8-methyl-3, 8-diazabicyclo [3.2.1] octane (3):
The compound 4-bromo-1-methoxy-2-nitrobenzene (258.0 mg,1.11 mmol) was dissolved in anhydrous 1, 4-dioxane (3 mL) and 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (41.5 mg,0.10 mmol) and tris (dibenzylideneacetone) dipalladium (92.5 mg,0.10 mmol) were added sequentially. Sodium tert-butoxide (194.1 mg,2.02 mmol) and 8-methyl-3, 8-diaza-bicyclo [3.2.1] are added under nitrogen protection]Octane hydrochloride (200.0 mg,1.00 mmol), and the reaction was stirred at 100℃for 2 hours. After the reaction was completed, the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness. Purification by column chromatography on silica gel (dichloromethane: methanol=10:1) afforded a brown oil (197.0 mg). LC-MS: [ M+H ]] + :278.1。
Synthesis of 2-methoxy-5- ((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1] oct-3-yl) aniline (4):
the compound (1R, 5S) -3- (4-methoxy-3-nitrophenyl) -8-methyl-3, 8-diazabicyclo [3.2.1]Octane (197.0 mg,0.71 mmol) was dissolved in methanol (5 mL), palladium on carbon (30.0 mg,10% w/w) was added under nitrogen, and replaced 3 times with hydrogen. The reaction solution was stirred at room temperature for 4 hours, then filtered through celite, the cake was washed twice with a small amount of methanol, and the filtrate was concentrated to give a brown yellow solid (160.0 mg). LC-MS: [ M+H ] ] + :248.2。
Synthesis of 5' - (5-fluoro-2- ((2-methoxy-5- ((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1] oct-3-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindolin ] -3' -one (DP-01-261):
the compound 2-methoxy-5- ((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1]Octane-3-yl) aniline (150.0 mg,0.61 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindole)]3' -Ketone (175.7 mg,0.61 mmol), tris (dibenzylideneacetone) dipalladium (55.5 mg,0.06 mmol) and 1,1' -binaphthyl-2, 2' -bisdiphenylphosphine (75.5 mg,0.12 mmol) were placed in anhydrous 1, 4-dioxane (5 mL), nitrogenCesium carbonate (494.0 mg,1.52 mmol) was added under protection and the reaction was stirred for 4 hours at 100 ℃. After the reaction was completed, the reaction mixture was diluted with water (20 mL) and extracted with methylene chloride (30 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The mixture was purified by preparative separation to give a yellow solid (32.6 mg). 1 H NMR(400MHz,DMSO-d6)δ8.91(s,1H),8.65(d,J＝3.2Hz,1H),8.30(s,1H),8.23(d,J＝8.0Hz,1H),8.18(s,1H),7.85(d,J＝2.8Hz,1H),7.48(d,J＝8.0Hz,1H),6.91(d,J＝8.8Hz,1H),6.47(dd,J＝8.8,2.8Hz,1H),3.79(s,3H),3.28–3.25(m,4H),2.85(d,J＝8.8Hz,2H),2.27(s,3H),1.94(d,J＝8.8Hz,2H),1.68–1.66(m,2H),1.55(s,4H)；LC-MS:[M+H] + :501.1。
EXAMPLE 68 Synthesis of Compound DP-01-222
Synthesis of tert-butyl 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3-methyl-1-oxoisoindoline-2-carboxylate (1):
the compound 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (dimethylphosphoryl) -3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (200.0 mg,0.44 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (97.0 mg,0.44 mmol), palladium acetate (9.0 mg,0.04 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (24.0 mg,0.04 mmol) and cesium carbonate (429.0 mg,1.32 mmol) were placed in 1, 4-dioxane (3 ml) under nitrogen atmosphere and heated to 100℃for reaction for 16 hours. After the reaction mixture was cooled to room temperature, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (30 mL. Times.3). The combined organic phases were washed with saturated brine (80 mL. Times.2), dried over anhydrous sodium sulfate, and concentrated by filtration to give the crude product as a yellow oil. The crude product was purified by reverse phase chromatography (10% methanol in dichloromethane) to give a yellow solid (120.0 mg). LC-MS: [ M+H ] ] + :639.2。
Synthesis of 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3-methylisoindol-1-one (DP-01-222):
the compound 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (100.0 mg,0.16 mmol), trifluoroacetic acid (0.3 mL) was dissolved in dichloromethane (1 mL) and reacted at room temperature for 1 hour. The reaction solution was directly concentrated and purified by preparative high performance liquid chromatography to give a yellow solid (45.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.92(s,1H),8.68(d,J＝3.2Hz,1H),8.33–8.32(m,3H),7.74(d,J＝2.4Hz,1H),6.93(d,J＝8.8Hz,1H),6.63(dd,J＝8.8,2.4Hz,1H),5.08(q,J＝6.4Hz,1H),3.79(s,3H),3.01(s,4H),2.44(s,4H),2.20(s,3H),1.80(dd,J＝13.2,3.2Hz,6H),1.59(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :539.3。
EXAMPLE 69 Synthesis of Compound DP-01-224
Synthesis of 7- (benzylthio) -5-bromo-1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (1):
5-bromo-7-iodo-1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (500.0 mg,1.07 mmol) and benzyl mercaptan (146.6 mg,1.18 mmol) were dissolved in 1, 4-dioxane (10 mL), and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (124.1 mg,0.21 mmol), tris (dibenzylideneacetone) dipalladium (196.5 mg,0.21 mmol) and triethylamine (0.45 mL) were added sequentially. Stirred at 50℃for 2 hours. The reaction solution was cooled to room temperature and concentrated to dryness. The crude product was purified by silica gel column chromatography (0-5% methanol/dichloromethane) to give a yellow oil (350.0 mg). LC-MS: [ M ] t Bu+H] + :406.1。
Synthesis of 5-bromo-1, 1-dimethyl-3-oxo-7-sulfamoyl isoindoline-2-carboxylic acid tert-butyl ester (2):
7- (Benzylthio) -5-bromo-1, 1-dimethyl-3-oxoisoindoline-2-carboxylic acid tert-butyl ester (350.0 mg,0.76 mmol) was dissolved in acetonitrile (5 mL), and acetic acid (0.1 mL) and water (0.2 mL) were added, followed by addition of 1, 3-dichloro-5, 5-dimethylhydantoin (298.2 mg,1.51 mmol) in portions. The reaction was stirred at room temperature for 30 minutes. Then, the reaction mixture was slowly dropped into 25% aqueous ammonia (2 mL),stirring was then continued for 0.5 hours at room temperature. After completion of the reaction, the reaction mixture was diluted with water (20 mL) and extracted with methylene chloride (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (0-50% ethyl acetate/petroleum ether) to give a yellow solid (260.0 mg). LC-MS: [ M ] t Bu+H] + :363.2。
Synthesis of 1, 1-dimethyl-3-oxo-7-sulfamoyl-5-boronic acid pinacol ester isoindoline-2-carboxylic acid tert-butyl ester (3):
the compound 5-bromo-1, 1-dimethyl-3-oxo-7-sulfamoyl isoindoline-2-carboxylic acid tert-butyl ester (190.0 mg,0.45 mmol), pinacol biborate (138.0 mg,0.54 mmol), [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (33.2 mg,0.046 mmol) and potassium acetate (66.7 mg,0.68 mmol) were suspended in anhydrous dioxane (5 mL) and stirred at 90℃for 1 hour under nitrogen. After completion of the reaction, water (10 mL) was added to the system, followed by extraction with ethyl acetate (50 mL. Times.3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (0-5% methanol/dichloromethane) to give a grey solid (150.0 mg). LC-MS: [ M ] t Bu+H] + :411.1。
Synthesis of 5- (2-chloro-5-fluoropyrimidin-4-yl) -1, 1-dimethyl-3-oxo-7-sulfonylisoindoline-2-carboxylic acid tert-butyl ester (4):
the compound 1, 1-dimethyl-3-oxo-7-sulfamoyl-5-boronic acid pinacol ester isoindoline-2-carboxylic acid tert-butyl ester (150.0 mg,0.32 mmol), 2, 4-dichloro-5-fluoropyrimidine (80.6 mg,0.48 mmol), potassium phosphate (204.8 mg,0.97 mmol) and [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (47.1 mg,0.06 mmol) was suspended in anhydrous 1, 4-dioxane (3 mL) and water (0.3 mL) and the reaction was stirred at 100deg.C under nitrogen for 1 hour. After the reaction is finished, the reaction liquid is directly concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (0-5% methanol/dichloromethane) to give a brown solid (35.0 mg). LC-MS: [ M ] t Bu+H] + :415.1。
Synthesis of 5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -1, 1-dimethyl-3-oxo-7-sulfonylisoindoline-2-carboxylic acid tert-butyl ester (5):
to a solution of tert-butyl 5- (2-chloro-5-fluoropyrimidin-4-yl) -1, 1-dimethyl-3-oxo-7-sulfonylisoindoline-2-carboxylate (35.0 mg,0.07 mmol) in 1, 4-dioxane (2 mL) was added successively 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (22.6 mg,0.10 mmol), 1 '-binaphthyl-2, 2' -diphenylphosphine (42.3 mg,0.07 mmol), tris (dibenzylideneacetone) dipalladium (62.2 mg,0.07 mmol) and cesium carbonate (66.4 mg,0.20 mmol), and heated to 100℃and stirred for 1 hour. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was chromatographed on silica gel (0-20% methanol/dichloromethane) to give a black solid (20.0 mg). LC-MS: [ M+H ] ] + :656.2。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3, 3-dimethyl-1-oxoisoindoline-4-sulfonamide (DP-01-224):
5- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -1, 1-dimethyl-3-oxo-7-sulfonylisoindoline-2-carboxylic acid tert-butyl ester (20.0 mg,0.03 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added thereto for 1 hour at room temperature. At the end of the reaction, the reaction mixture was concentrated in vacuo and the crude was purified by preparative chromatography to give a white solid (5.0 mg). 1 HNMR(400MHz,DMSO-d6)δ9.13(s,1H),8.78(s,1H),8.73(d,J＝3.2Hz,1H),8.42(s,1H),8.30(s,1H),7.83(d,J＝2.9Hz,1H),6.94(d,J＝8.9Hz,1H),6.62(dd,J＝8.9,2.8Hz,1H),3.81(s,3H),3.05–3.02(m,4H),2.45–2.43(m,4H),2.20(s,3H),1.72(s,6H)；LC-MS:[M+H] + :556.3。
EXAMPLE 70 Synthesis of Compounds DP-01-228
Synthesis of tert-butyl 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3-methyl-1-oxoisoindoline-2-carboxylate (1):
the compound 6- (2-chloro-5-fluoropyrimidin-4-yl) -4- (dimethylphosphoryl) -3-methyl-1-oxo-isoindoline-2-carboxylic acid tert-butyl ester200.0mg,0.44 mmol), 5- (4-methylpiperazin-1-yl) -2-trifluoromethoxyaniline (146.0 mg,0.53 mmol), palladium acetate (9.0 mg,0.04 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (25.0 mg,0.04 mmol), cesium carbonate (429.0 mg,1.32 mmol) were dissolved in 1, 4-dioxane (3 ml), and the mixture was reacted at 100℃under nitrogen atmosphere for 3 hours. After the reaction solution was cooled, the sample was purified by a column chromatography (0% -10% methanol/dichloromethane) to obtain a yellow solid (130.0 mg). LC-MS: [ M+H ] ] + :693.3。
Synthesis of 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3-methylisoindol-1-one (DP-01-228):
after dissolving the compound 4- (dimethylphosphoryl) -6- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (100.0 mg,0.14 mmol), trifluoroacetic acid (0.3 ml) in dichloromethane (1 ml), the mixture was reacted at room temperature for 1 hour. After the reaction solution was dried by spin-drying, it was purified by preparation (42.0 mg). 1 H NMR(400MHz,DMSO)δ9.21(s,1H),8.90(s,1H),8.66(d,J＝3.2Hz,1H),8.31–8.28(m,2H),7.33(d,J＝2.8Hz,1H),7.21(d,J＝8.8Hz,1H),6.78(dd,J＝8.8,2.8Hz,1H),5.06(q,J＝6.4Hz,1H),3.17–3.10(m,4H),2.46–2.40(m,4H),2.20(s,3H),1.77(dd,J＝13.2,2.0Hz,6H),1.57(d,J＝6.4Hz,3H)；LC-MS:[M+H] + :593.3。
EXAMPLE 71 Synthesis of Compound DP-01-235
Synthesis of 1- (tert-butyl) 3-methyl-4-methylpiperazine-1, 3-dicarboxylic acid ester (2):
compound 1- (tert-butyl) 3-methylpiperazine-1, 3-dicarboxylic acid ester (1.0 g,4.09 mmol), aqueous formaldehyde solution (647.4 mg,8.20mmol, 38%) was dissolved in acetonitrile (10 ml) and water (2 ml), and sodium cyanoborohydride (516.6 mg,8.22 mmol) was slowly added to the reaction solution and stirred at room temperature for 2 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL. Times.3). The combined organic phases were washed with saturated brine (80 mL. Times.2), anhydrous sodium sulfateDrying, filtering and concentrating to obtain colorless oily crude product. The crude product was purified by column chromatography (0-10% ethyl acetate/petroleum ether) to give a colorless oil (560.0 mg). LC-MS: [ M+H ] ] + :259.3。
Synthesis of 3- (2-hydroxypropyl-2-yl) -4-methylpiperazine-1-carboxylic acid tert-butyl ester (3):
the compound 1- (tert-butyl) 3-methyl-4-methylpiperazine-1, 3-dicarboxylic acid ester (540.0 mg,2.09 mmol) was dissolved in methylene chloride (10 mL), and after cooling the reaction solution to-65℃methyl magnesium bromide (4.6 mL,4.60mmol,1 mol/L) was slowly added dropwise. After the completion of the dropwise addition, the reaction mixture was allowed to stand at room temperature for 2 hours. The reaction mixture was diluted with 50mL of aqueous ammonium chloride, and extracted with methylene chloride (50 mL. Times.2). The combined organic phases were washed with saturated brine (100 mL. Times.2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=2:1) to give a colorless oil (390.0 mg). 1 H NMR(400MHz,DMSO)δ4.26(s,1H),3.75(d,J＝11.2Hz,1H),3.60(d,J＝12.8Hz,1H),2.92(s,1H),2.80–2.74(m,1H),2.38(s,3H),2.34–2.30(m,1H),2.03(dd,J＝9.2,4.0Hz,1H),1.40(s,9H),1.09(d,J＝8.4Hz,6H)。
Synthesis of 2- (1-methylpiperazin-2-yl) propan-2-ol (4):
the compound 3- (2-hydroxy-prop-2-yl) -4-methylpiperazine-1-carboxylic acid tert-butyl ester (370.0 mg,1.43 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (1 mL) was added, and after stirring at room temperature for 1 hour, the reaction solution was dried by spinning. Methyl tert-butyl ether was slurried and filtered to give a crude white product (200.0 mg) which was used directly in the next reaction. LC-MS: [ M+H ]] + :159.1。
Synthesis of 2- (4- (4-methoxy-3-nitrophenyl) -1-methylpiperazin-2-yl) propan-2-ol (5):
the compound 2- (1-methylpiperazin-2-yl) propan-2-ol (150.0 mg,0.95 mmol), 4-bromo-1-methoxy-2-nitrobenzene (263.2 mg,1.13 mmol), tris (dibenzylideneacetone) dipalladium (91.5 mg,0.10 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (47.6 mg,0.10 mmol), sodium t-butoxide (182.4 mg,1.90 mmol) was dissolved in toluene (3 mL) and reacted at 90℃for 5 hours. The reaction mixture was cooled, diluted with water (30 mL), and extracted with ethyl acetate (30 mL. Times.3). The combined organic phases were washed with saturated aqueous sodium chloride (50 mL), dried over anhydrous sodium sulfate Filtering and concentrating to obtain a crude product. The crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=1:1) to give a colorless oil (45.0 mg). LC-MS: [ M+H ]] + :310.1。
Synthesis of 2- (4- (3-amino-4-methoxyphenyl) -1-methylpiperazin-2-yl) propan-2-ol (6):
the compound 2- (4- (4-methoxy-3-nitrophenyl) -1-methylpiperazin-2-yl) propan-2-ol (40.0 mg,0.13 mmol), iron powder (36.2 mg,0.65 mmol), ammonium chloride (34.8 mg,0.65 mmol) were dissolved in methanol (1 mL) and water (0.2 mL), reacted at 80℃for 3 hours, the reaction solution was cooled, filtered and concentrated to give a brown solid (35.0 mg). LC-MS: [ M+H ]] + :280.1。
Synthesis of 5' - (5-fluoro-2- ((5- (3- (2-hydroxypropan-2-yl) -4-methylpiperazin-1-yl) -2-methoxyphenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-235):
the compound 2- (4- (3-amino-4-methoxyphenyl) -1-methylpiperazin-2-yl) propan-2-ol (30.0 mg,0.11 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropan-1, 1' -isoindoline]3' -Ketone (31.8 mg,0.11 mmol), tris (dibenzylideneacetone) dipalladium (9.6 mg,0.01 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (6.9 mg,0.01 mmol), cesium carbonate (71.5 mg,0.22 mmol) were dissolved in 1, 4-dioxane (1 mL) and heated to 100deg.C under nitrogen for 16 hours. The reaction mixture was cooled, diluted with water (30 mL), and extracted with ethyl acetate (30 mL. Times.3). The combined organic phases were washed with saturated brine (80 mL. Times.2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by prep. to give a white solid (14.1 mg). 1 H NMR(400MHz,DMSO-d6)δ8.89(s,1H),8.64(d,J＝3.6Hz,1H),8.28(s,1H),8.22–8.21(m,2H),7.84(d,J＝2.8Hz,1H),7.49(d,J＝8.0Hz,1H),6.93(d,J＝8.8Hz,1H),6.55(dd,J＝8.8,2.8Hz,1H),4.29(s,1H),3.80(s,3H),3.37(s,2H),2.85(d,J＝12.0Hz,1H),2.73–2.70(m,1H),2.46(d,J＝4.8Hz,2H),2.40(s,3H),2.19(dd,J＝9.6,3.2Hz,1H),1.54(s,4H),1.10(s,3H),1.02(s,3H).LC-MS:[M+H] + :533.3。
EXAMPLE 72 Synthesis of Compounds DP-01-240, DP-01-240A, DP-01-240B
Synthesis of tert-butyl 4- (3-bromo-4-methoxyphenyl) -2- (hydroxymethyl) piperazine-1-carboxylate (2):
to a solution of 2-bromo-4-iodo-1-methoxybenzene (4.1 g,13.10 mmol) in dimethyl sulfoxide (20 mL) was added tert-butyl 2- (hydroxymethyl) piperazine-1-carboxylate (3.4 g,15.72 mmol), cesium carbonate (5.1 g,15.65 mmol), cuprous iodide (0.5 g,2.63 mmol) and L-proline (0.3 g,2.61 mmol), and the mixture was stirred under nitrogen at 90℃for 1 hour. After completion of the reaction, the reaction mixture was diluted with 200mL of water and extracted with ethyl acetate (200 mL. Times.3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=8:1) to give a yellow oily liquid (1.2 g). LC-MS: [ M+H ]] + :401.1,403.1。
Synthesis of (4- (3-bromo-4-methoxyphenyl) piperazin-2-yl) methanol (3):
tert-butyl 4- (3-bromo-4-methoxyphenyl) -2- (hydroxymethyl) piperazine-1-carboxylate (1.1 g,2.74 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (4 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was directly concentrated to give an oil (815.0 mg) which was directly used in the next reaction. LC-MS: [ M+H ] ] + :301.1,303.1。
Synthesis of (4- (3-bromo-4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (4):
(4- (3-bromo-4-methoxyphenyl) piperazin-2-yl) methanol (330.0 mg,1.10 mmol) was dissolved in water (2 mL) and acetonitrile (5 mL), then a 38% aqueous formaldehyde solution (0.5 mL) and acetic acid (30.0 mg,0.5 mmol) were added to the reaction solution, and after stirring at room temperature for 1 hour, sodium triacetoxyborohydride (694.9 mg,3.28 mmol) was added. The suspension was then stirred for 3 hours, quenched with saturated aqueous sodium bicarbonate (20 mL) and extracted with ethyl acetate (100 mL. Times.2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give the crude product. The crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (238.0 mg). LC-MS: [ M+H ]] + :315.1,317.1。
Synthesis of (4- (3- ((diphenylmethylene) amino) -4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (5):
to a solution of (4- (3-bromo-4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (500.0 mg,1.59 mmol) in 1, 4-dioxane (3 mL) was added benzophenone imine (575.0 mg,3.17 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (197.6 mg,0.32 mmol), tris (dibenzylideneacetone) dipalladium (290.5 mg,0.32 mmol) and cesium carbonate (1550.5 mg,4.76 mmol) in this order, and the mixture was heated to 100℃under nitrogen atmosphere and stirred for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (320.0 mg). LC-MS: [ M+H ] ] + :416.3。
Synthesis of (4- (3-amino-4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (6):
(4- (3- ((diphenylmethylene) amino) -4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (320.0 mg,0.77 mmol) was dissolved in 1, 4-dioxane (3 mL) and 2M dioxane hydrochloride solution (3 mL) was added, and the reaction solution was stirred at room temperature for 1 hour. After the completion of the reaction, the reaction mixture was concentrated in vacuo to give a pale yellow oil (110.0 mg) which was used directly in the next reaction. LC-MS: [ M+H ]] + :252.3。
Synthesis of 5' - (5-fluoro-2- ((5- (3- (hydroxymethyl) -4-methylpiperazin-1-yl) -2-methoxyphenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-240):
to a solution of (4- (3-amino-4-methoxyphenyl) -1-methylpiperazin-2-yl) methanol (100.0 mg,0.40 mmol) in 1, 4-dioxane (5 mL) was added successively 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (138.1 mg,0.48 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (49.6 mg,0.08 mmol), tris (dibenzylideneacetone) dipalladium (36.4 mg,0.04 mmol) and cesium carbonate (388.9 mg,1.19 mmol) were heated to 100℃and stirred for 4 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by prep. HPLC to give a yellow solid (22.8 mg). 1 HNMR(400MHz,DMSO-d6)δ8.90(s,1H),8.65(d,J＝3.6Hz,1H),8.30(s,1H),8.27–8.24(m,2H),7.88(s,1H),7.51(d,J＝8.2Hz,1H),6.95(d,J＝8.8Hz,1H),6.61(d,J＝8.8Hz,1H),4.57(s,1H),3.81(s,3H),3.62(s,1H),3.52(d,J＝10.8Hz,1H),3.42–3.38(m,1H),3.28–3.27(m,1H),2.84(s,1H),2.73(s,1H),2.49–2.42(m,1H),2.33–2.28(m,5H),1.55(s,4H)；LC-MS:[M+H] + :505.2。
The racemate DP-01-240 was purified by SFC (column CHIRALPAK AD X4.6 mm 5 μm; mobile phase: A phase: 50% n-Hexane (0.05% DEA); B phase: etOH; gradient: B%:50%,10 min) to give white solid DP-01-240A and white solid DP-01-240B.
DP-01-240A：
Chiral analysis conditions: chromatographic column: CHIRALPAK AD 100 x 4.6mm 5 μm, mobile phase: phase A: 50% (0.05% DEAN-Hexane), phase B: etOH; gradient: b%:50% retention time 6.084 min, ee% = 98.65%.
1 H NMR(400MHz,DMSO)δ8.96(s,1H),8.71(d,J＝3.5Hz,1H),8.36(s,1H),8.31–8.30(m,2H),7.93(d,J＝2.9Hz,1H),7.57(d,J＝8.0Hz,1H),7.00(d,J＝8.9Hz,1H),6.66(dd,J＝8.9,3.0Hz,1H),4.60(s,1H),3.87(s,3H),3.66(s,1H),3.57(d,J＝11.9Hz,1H),3.45–3.44(m,2H),2.86(d,J＝11.2Hz,1H),2.75(d,J＝14.0Hz,1H),2.50(s,1H),2.38(d,J＝13.2Hz,1H),2.31(s,3H),2.21(s,1H),1.61(s,4H)；LC-MS:[M+H] + :505.3。
DP-01-240B：
Chiral analysis conditions: chromatographic column: CHIRALPAK AD 100 x 4.6mm 5 μm, mobile phase: phase A: 50% (0.05% DEAN-Hexane), phase B: etOH; gradient: b%:50% retention time 7.531 min, ee% = 94.29%.
1 H NMR(400MHz,DMSO-d6)δ8.90(s,1H),8.65(d,J＝3.6Hz,1H),8.30(s,1H),8.28–8.24(m,2H),7.87(d,J＝2.7Hz,1H),7.51(d,J＝8.1Hz,1H),6.94(d,J＝8.9Hz,1H),6.60(dd,J＝8.8,2.9Hz,1H),4.53(s,1H),3.81(s,3H),3.62(d,J＝10.2Hz,1H),3.51(d,J＝11.3Hz,1H),3.41–3.36(m,2H),2.80(d,J＝11.3Hz,1H),2.70(d,J＝14.3Hz,1H),2.45(d,J＝11.2Hz,1H),2.34–2.29(m,1H),2.25(s,3H),2.15(s,1H),1.55(s,4H)；LC-MS:[M+H] + :505.2。
EXAMPLE 73 Synthesis of Compounds DP-01-243
Synthesis of 4-bromo-1-cyclopropyl-2-nitrobenzene (2):
the compound 4-bromo-1-iodo-2-nitrobenzene (2.5 g,7.62 mmol), cyclopropylboronic acid (1.0 g,11.64 mmol), palladium acetate (172.9 mg,0.77 mmol), tricyclohexylphosphine (215.9 mg,0.77 mmol), potassium phosphate (4.9 g,23.08 mmol) was dissolved in toluene (30 ml) and water (3 ml), and the mixture was heated to 100℃under nitrogen protection to react for 3 hours. After the reaction solution was cooled, the reaction solution was diluted with water (300 mL) and extracted with ethyl acetate (300 mL. Times.2). The organic phase was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (0% -15% ethyl acetate/petroleum ether) to give a yellow solid (730.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.09(d,J＝2.0Hz,1H),7.78(dd,J＝8.4,2.0Hz,1H),7.20(d,J＝8.4Hz,1H),2.14(tt,J＝8.4,5.2Hz,1H),1.05–0.99(m,2H),0.79–0.71(m,2H)。
Synthesis of 1- (4-cyclopropyl-3-nitrophenyl) -4-methylpiperazine (3):
the compound 4-bromo-1-cyclopropyl-2-nitrobenzene (700.0 mg,2.89 mmol), N-methylpiperazine (433.0 mg,4.32 mmol), tris (dibenzylideneacetone) dipalladium (265.0 mg,0.29 mmol), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (237.0 mg,0.58 mmol), cesium carbonate (2.8 g,8.59 mmol) were dissolved in 1, 4-dioxane (7 ml) and reacted at 100℃for 6 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL. Times.2). The organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by silica gel column chromatography (0-40% ethyl acetate/petroleum ether) to give a yellow oil (330.0 mg). LC-MS: [ M+H ]] + :262.1。
Synthesis of 2-cyclopropyl-5- (4-methylpiperazin-1-yl) aniline (4):
the compound 1- (4-cyclopropyl-3-nitrophenyl) -4-methylpiperazine (100.0 mg,0.38 mmol), iron powder (63.0 mg,1.13 mmol), ammonium chloride (161.0 mg,3.01 mmol) were dissolved in methanol (1 mL) and water (0.2 mL), reacted at 80℃for 3 hours, the reaction solution was cooled, filtered, and the cake was ultrasonically filtered 3 times using methanol washing, and the filtrates were concentrated to give a black crude product (80.0 mg)) Directly used in the next step. LC-MS: [ M+H ]] + :232.1。
Synthesis of 5' - (2- ((2-cyclopropyl-5- (4-methylpiperazin-1-yl) phenyl) amino) -5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-243):
The compound 2-cyclopropyl-5- (4-methylpiperazin-1-yl) aniline (70.0 mg,0.30 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (87.0 mg,0.30 mmol), tris (dibenzylideneacetone) dipalladium (28.8 mg,0.03 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (37.3 mg,0.06 mmol), cesium carbonate (293.0 mg,0.90 mmol) were placed in 1, 4-dioxane (1 mL) and heated to 100deg.C under nitrogen for 16 hours. The reaction mixture was cooled, diluted with water (30 mL), and extracted with ethyl acetate (30 mL. Times.3). The organic phase was washed with saturated brine (80 mL. Times.2), dried over anhydrous sodium sulfate, and concentrated by filtration. The crude product was purified by preparative high performance liquid chromatography to give a white solid (15.2 mg). 1 H NMR(400MHz,DMSO-d6)δ8.87(s,1H),8.70(s,1H),8.60(d,J＝3.6Hz,1H),8.30(s,1H),8.21(d,J＝8.0Hz,1H),7.47(d,J＝8.0Hz,1H),7.41(s,1H),6.88(d,J＝8.4Hz,1H),6.65(d,J＝8.4Hz,1H),3.10(s,4H),2.47(s,4H),2.23(s,3H),1.94–1.86(m,1H),1.54(s,4H),0.82(d,J＝8.0Hz,2H),0.52(d,J＝4.4Hz,2H)；LC-MS:[M+H] + :485.3。
EXAMPLE 74 Synthesis of Compounds DP-01-256
Synthesis of 6-bromo-3-methyl-4- (methylthio) -1-oxoisoindoline-2-carboxylic acid tert-butyl ester (1):
the compound 6-bromo-4-iodo-3-methyl-1-oxoisoindoline-2-carboxylic acid tert-butyl ester (1.6 g,3.54 mmol), sodium methyl mercaptide (744.2 mg,10.62 mmol), cuprous iodide (67.4 mg,0.35 mmol) and L-proline sodium salt (98.5 mg,0.72 mmol) were dissolved in dimethyl sulfoxide (16.0 mL). The reaction system was stirred at 90℃for 3 hours under nitrogen protection. After completion of the reaction, the reaction was quenched by addition of water (20.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated Filtering and concentrating to obtain crude product. Column chromatography (DCM: meoh=20:1) afforded a yellow solid (900.0 mg). LC-MS: [ M+H-Boc ]] + :272.9,274.9。
Synthesis of 6-bromo-3-methyl-4- (methylthio) isoindolin-1-one (2):
the compound 6-bromo-3-methyl-4- (methylsulfanyl) -1-oxoisoindoline-2-carboxylic acid tert-butyl ester (900.0 mg,2.42 mmol) was dissolved in dichloromethane (10.0 mL) and trifluoroacetic acid (1.0 mL) was slowly added dropwise. The reaction system was stirred at room temperature for 1 hour and then concentrated to give a crude product. Purification by column chromatography (DCM: meoh=20:1) afforded a yellow solid (530.0 mg). LC-MS: [ M+H ]] + :272.0,274.0。
Synthesis of 6-bromo-3-methyl-4- (methylsulfonyl) isoindolin-1-one (3):
the compound 6-bromo-3-methyl-4- (methylthio) isoindolin-1-one (528.0 mg,1.94 mmol) and potassium hydrogen peroxymonosulfate complex (3.58 g,10.34 mmol) were dissolved in tetrahydrofuran (5 mL) and water (5 mL), and the reaction system was stirred at room temperature for 12 hours. After completion of the reaction, the reaction was quenched with water (20.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Purification by column chromatography (DCM: meoh=20:1) afforded a yellow solid (300.0 mg). LC-MS: [ M+H ]] + :303.9,305.9。
Synthesis of (1-methyl-7- (methylsulfonyl) -3-oxoisoindolin-5-yl) boronic acid (4):
The compound 6-bromo-3-methyl-4- (methylsulfonyl) isoindolin-1-one (300.0 mg,0.99 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (72.0 mg,0.10 mmol), pinacol biborate (325.2 mg,1.28 mmol), potassium acetate (289.8 mg,2.95 mmol) were placed in 1, 4-dioxane (3.0 mL). The reaction system was stirred at 80℃for 1 hour under nitrogen protection. After the reaction was completed, the reaction solution was concentrated in vacuo to give a black solid (266.4 mg) which was directly used for the next reaction. LC-MS: [ M+H ]] + :270.1。
Synthesis of 6- (2-chloro-5-fluoropyrimidin-4-yl) -3-methyl-4- (methylsulfonyl) isoindolin-1-one (5):
(1-methyl-7- (methylsulfonyl) -3-oxoisoindolin-5-yl) boronic acid (266)4mg,0.99 mmol), 2, 4-dichloro-5-fluoropyrimidine (197.4 mg,1.18 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (72.0 mg,0.10 mmol), potassium phosphate (626.4 mg,2.95 mmol) were dissolved in a mixed solution of 1, 4-dioxane and water (3.3 mL,1, 4-dioxane: water=10:1). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Column chromatography (DCM: meoh=10:1) afforded a yellow solid (280.0 mg). LC-MS: [ M+H ] ] + :356.1,358.0。
Synthesis of 6- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -3-hydroxy-3-methyl-4- (methylsulfonyl) isoindolin-1-one (DP-01-256):
2-methoxy-5- (4-methylpiperazin-1-yl) aniline (34.2 mg,0.15 mmol), 6- (2-chloro-5-fluoropyrimidin-4-yl) -3-methyl-4- (methylsulfonyl) isoindolin-1-one (50.0 mg,0.14 mmol), tris (dibenzylideneacetone) dipalladium (12.9 mg,0.014 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (17.6 mg,0.028 mmol), cesium carbonate (91.9 mg,0.28 mmol) were placed in 1, 4-dioxane (1.0 mL). The reaction system was stirred at 90℃for 2 hours under nitrogen protection. After completion of the reaction, the reaction was quenched with water (10.0 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Yellow solid (14.0 mg) was obtained by preparative separation. 1 H NMR(400MHz,CDCl 3 )δ8.90(s,1H),8.86(s,1H),8.48(d,J＝2.8Hz,1H),8.24(d,J＝2.4Hz,1H),7.90(s,1H),6.84(d,J＝8.8Hz,1H),6.58(dd,J＝8.8,2.8Hz,1H),3.91(s,3H),3.33(s,3H),3.26–3.16(m,4H),2.69–2.56(m,4H),2.28(s,3H),2.13(s,3H)；LC-MS:[M+H] + :557.1。
EXAMPLE 75 Synthesis of Compound DP-01-267
Synthesis of 5-acetyl-2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (2):
the compound 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (214.9 mg,0.97 mmol), 5-acetyl-2- (2-chloro-5-fluoropyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] under nitrogen ]Pyrrole-4-one (300.0 mg,0.88 mmol), tris (dibenzylideneacetone) dipalladium (80.9 mg,0.088 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (110.0 mg,0.18 mmol), cesium carbonate (575.4 mg,1.77 mmol) were placed in 1, 4-dioxane (3.0 mL), and heated to 100℃to react for 2 hours. After the reaction was complete, the reaction was cooled to room temperature. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed successively with water (10 mL) and saturated aqueous sodium chloride (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. Purification by column chromatography (dichloromethane: methanol=10:1) afforded a yellow solid (65.4 mg). LC-MS: [ M+H ]] + :525.1。
Synthesis of 2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c ] pyrrol-4-one (DP-01-267):
the compound 5-acetyl-2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6, 6-dimethyl-5, 6-dihydro-4H-thieno [2,3-c]Pyrrole-4-one (50.0 mg,0.095 mmol) and sodium hydroxide (11.4 mg,0.28 mmol) were dissolved in a mixed solution of tetrahydrofuran (1.0 mL) and water (1.0 mL), and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL. Times.3). The combined organic phases were washed successively with water (10 mL) and saturated aqueous sodium chloride (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give crude product. The crude product was isolated as a yellow solid (8.0 mg). 1 H NMR(400MHz,DMSO-d6)δ8.38(dd,J＝4.8,2.8Hz,2H),8.01(d,J＝1.2Hz,1H),7.87(s,1H),6.84(d,J＝8.8Hz,1H),6.58(dd,J＝8.8,2.8Hz,1H),6.23(s,1H),3.90(s,3H),3.36–3.22(m,4H),2.71–2.63(m,4H),2.40(s,3H),1.70(s,6H)；LCMS:[M+H] + :483.0。
EXAMPLE 76 Synthesis of Compound DP-01-269
Synthesis of 5' - (2, 5-dichloropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (1):
spiro [1,1 '-isoindole ] as 5' -boronic acid pinacol ester]-3 '-one (200.0 mg,0.70 mmol), cesium carbonate (455.0 mg,1.40 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (56.7 mg,0.07 mmol), 2,4, 5-trichloropyrimidine (127.0 mg,0.69 mmol) were dissolved in 1, 4-dioxane and water (10 mL, 5:1), and stirred at 100deg.C under nitrogen for 6 hours. The reaction mixture was cooled to room temperature, poured into water (20 mL), and extracted with ethyl acetate (15 mL. Times.3). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (ethyl acetate: petroleum ether=2:1) to give a yellow solid (150.0 mg). LC-MS: [ M+H ]] + :306.2,308.1.5'- (5-chloro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-synthesis of 3' -ketone (DP-01-269):
the compound 5'- (2, 5-dichloropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]-3' -Ketone (150.0 mg,0.49 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (109.0 mg,0.49 mmol), cesium carbonate (478.0 mg,1.47 mmol), tris (dibenzylideneacetone) dipalladium (47.4 mg,0.052 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (64.5 mg,0.10 mmol) were dissolved in toluene (10 mL) and stirred under nitrogen for 6 hours at 100 ℃. The reaction mixture was cooled to room temperature, poured into water (20 mL), and extracted with ethyl acetate (15 mL. Times.3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated and chromatographed on silica gel (dichloromethane: methanol=10:1) to give the crude product. The crude product was purified by prep. to give a white solid (28.5 mg). 1 H NMR(400MHz,DMSO-d6)δ8.87(s,1H),8.62(s,1H),8.37(s,1H),8.11(d,J＝1.0Hz,1H),8.03(dd,J＝8.0,1.6Hz,1H),7.84(d,J＝2.2Hz,1H),7.45(d,J＝8.1Hz,1H),6.92(d,J＝8.9Hz,1H),6.60(dd,J＝8.9,2.9Hz,1H),3.79(s,3H),3.01–2.98(m,4H),2.43–2.41(m,4H),2.20(s,3H),1.53(d,J＝2.1Hz,4H).LC-MS:[M+H] + :491.5。
EXAMPLE 77 Synthesis of Compounds DP-01-245
Synthesis of methyl 5-chloro-2- (chloromethyl) nicotinate (2):
the compound methyl 5-chloro-2-methylnicotinate (3.1 g,16.70 mmol), trichloroisocyanuric acid (11.6 g,49.91 mmol), benzamide (0.2 g,1.65 mmol) were dissolved in chloroform (50 mL) and stirred at 65℃for 40 hours. To the reaction solution was added 50mL of a saturated sodium carbonate solution, followed by stirring for 20 minutes, and extraction with methylene chloride (20 mL. Times.3) was performed. The organic phases were combined and dried over anhydrous sodium sulfate, concentrated by filtration, and purified by column chromatography (petroleum ether: ethyl acetate=150:1 to 80:1) to give a colorless oil (2.1 g). LCMS [ M+H ]] + :220.0,222.0。
Synthesis of 3-chloro-6- (4-methoxybenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (3):
the compound methyl 5-chloro-2- (chloromethyl) nicotinate (2.0 g,9.09 mmol) and 4-methoxybenzylamine (1.5 g,10.93 mmol) were dissolved in acetonitrile (10 mL), N-diisopropylethylamine (2.35 g,18.18 mmol) was slowly added dropwise and stirred at room temperature for 16 hours. The reaction mixture was diluted with water (20 mL) and extracted with methylene chloride (20 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=4:1-2:1) to give a white solid compound (1.2 g). 1 H NMR(400MHz,DMSO)δ8.79(d,J＝2.4Hz,1H),8.24(d,J＝2.0Hz,1H),7.25(d,J＝8.8Hz,2H),6.91(d,J＝8.4Hz,2H),4.68(s,2H),4.40(s,2H),3.73(s,3H)。
Synthesis of 3-chloro-6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (4):
the compound 3-chloro-6- (4-methoxybenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridin-5-one (1.2 g,4.16 mmol) was dissolved in dry tetrahydrofuran (20 mL), and lithium bis (trimethylsilylamide) (16.6 mL,16.6mmol,1 mol/L) was slowly added dropwise at 0deg.C and stirred at room temperature for 1 hour, methyl iodide (2.36 g,16.63 mmol) was slowly added dropwise and stirring continuedShould be 1 hour. The reaction was quenched with 10mL of saturated ammonium chloride and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=100:1 to 30:1) to give a white solid compound (580.0 mg). LCMS [ M+H ]] + :317.1,319.1。
Synthesis of (6- (4-methoxybenzyl) -7, 7-dimethyl-5-oxo-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-3-yl) boronic acid (5):
the compound 3-chloro-6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridin-5-one (580.0 mg,1.83 mmol), pinacol biborate (781.6 mg,3.08 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (195.6 mg,0.41 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2 ',4',6' -triisopropyl-1, 1' -biphenyl) (2 ' -amino-1, 1' -biphenyl-2-yl) palladium (II) (173.7 mg,0.21 mmol), potassium acetate (604.1 mg,6.16 mmol) were dissolved in dioxane (10 mL) and reacted under nitrogen at 80℃for 5 hours. The reaction mixture was cooled, diluted with water (10 mL), and extracted with ethyl acetate (15 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated to give a crude black product (350.0 mg) which was used directly in the next reaction. LCMS [ M+H ] ] + :327.1。
Synthesis of 3- (2-chloro-5-fluoropyrimidin-4-yl) -6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (6):
the compound (6- (4-methoxybenzyl) -7, 7-dimethyl-5-oxo-6, 7-dihydro-5H-pyrrolo [3, 4-b)]Pyridine-3-yl) boronic acid (350.0 mg,1.07 mmol), 2, 4-dichloro-5-fluoropyrimidine (268.8 mg,1.61 mmol), potassium phosphate (683.3 mg,3.22 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium (II) dichloride (78.5 mg,0.11 mmol) was dissolved in dioxane (8 mL) and water (2 mL). The reaction was carried out at 80℃for 5 hours under nitrogen protection. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, and the crude product obtained by filtration concentration was purified by column chromatography (petroleum ether: ethyl acetate=5:1 to 3:1) to give a white solid (250.0 mg). LCMS [ M+H ]] + :413.1,415.1。
Synthesis of 3- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (7):
the compound 3- (2-chloro-5-fluoropyrimidin-4-yl) -6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridin-5-one (250.0 mg,0.61 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (147.5 mg,0.67 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (75.9 mg,0.12 mmol), tris (dibenzylideneacetone) dipalladium (55.9 mg,0.061 mmol), cesium carbonate (589.3 mg,1.81 mmol) were dissolved in dioxane and reacted for 5 hours under nitrogen protection at 100 ℃. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated by filtration, and purified by column chromatography (dichloromethane: methanol=100:1 to 40:1) to give a white solid (240.0 mg). LCMS [ M+H ] ] + :598.4。
Synthesis of 3- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b ] pyridin-5-one (DP-01-245):
the compound 3- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6- (4-methoxybenzyl) -7, 7-dimethyl-6, 7-dihydro-5H-pyrrolo [3,4-b]Pyridin-5-one (240.0 mg,0.40 mmol) was dissolved in trifluoroacetic acid (3 mL) and trifluoromethanesulfonic acid (1 mL) and reacted at 60℃for 2.5 h. The reaction was concentrated to completion, 10mL of water was added and the pH of the solution was adjusted to alkaline with ammonia. Extracted with dichloromethane (15 mL x 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by prep. to give the compound as a yellow solid (40.2 mg). 1 H NMR(400MHz,DMSO)δ9.31(s,1H),9.07(s,1H),8.70(d,J＝3.2Hz,1H),8.50(s,1H),8.32(s,1H),7.86(d,J＝2.8Hz,1H),6.93(d,J＝8.8Hz,1H),6.60(dd,J＝9.2,3.2Hz,1H),3.79(s,3H),3.07–3.00(m,4H),2.46–2.41(m,4H),2.20(s,3H),1.51(s,6H).LCMS:[M+H] + :478.2。
EXAMPLE 78 Synthesis of Compounds DP-01-246
Synthesis of methyl 3- (bromomethyl) -6-chloropicolinate (2):
methyl 6-chloro-3-methylpyridine carboxylate (2.9 g,15.62 mmol) was dissolved in carbon tetrachloride (50 mL) and azobisisobutyronitrile (257.5 mg,1.57 mmol) and N-bromosuccinimide (2.8 g,15.73 mmol) were added under nitrogen. The reaction was stirred at 80℃for 16 hours, after the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate=8:1) to give a white solid (2.2 g). LC-MS: [ M+H ] ] + :264.1,266.1。
Synthesis of 2-chloro-6- (4-methoxybenzyl) -5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (3):
methyl 3- (bromomethyl) -6-chloropicolinate (2.2 g,8.32 mmol) and (4-methoxyphenyl) methylamine (1.4 g,10.20 mmol) were dissolved in tetrahydrofuran (30 mL), N-diisopropylethylamine (1.6 g,12.38 mmol) was slowly added and stirred at room temperature for 6 hours, quenched with saturated aqueous ammonium chloride (100 mL), extracted with ethyl acetate (200 mL. Times.2), the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=2:1) to give a white solid (1.8 g). LC-MS: [ M+H] + :289.1,291.1。
Synthesis of 2-chloro-6- (4-methoxybenzyl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (4):
2-chloro-6- (4-methoxybenzyl) -5, 6-dihydro-7H-pyrrolo [3,4-b]Pyridin-7-one (1.8 g,6.23 mmol), sodium hydride (550.0 mg,13.75mmol,60% w/w) were added to tetrahydrofuran (50 mL), stirred at room temperature for 1 hour, methyl iodide (2.0 g,14.09 mmol) was added, and stirring was continued at room temperature for 1 hour. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (150 mL. Times.2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration to give a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=1:1) to give a white solid (1.2 g). LC-MS: [ M+H ] ] + :317.1。
Synthesis of 6- (4-methoxybenzyl) -5, 5-dimethyl-2- (trimethylstannyl) -5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (5):
2-chloro-6- (4-methoxybenzyl) -5, 5-dimethyl1-5, 6-dihydro-7H-pyrrolo [3,4-b]Pyridin-7-one (310.0 mg,0.98 mmol), tetrakis (triphenylphosphine) palladium (116.0 mg,0.10 mmol) and hexamethylditin (490.0 mg,1.50 mmol) were dissolved in 1, 4-dioxane (10 mL) and heated to 100deg.C under nitrogen and stirred for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate=1:1) to give a gray solid (360.0 mg). LC-MS: [ M+H ]] + :447.1。
Synthesis of 2- (2-chloro-5-fluoropyrimidin-4-yl) -6- (4-methoxybenzyl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (6):
to 6- (4-methoxybenzyl) -5, 5-dimethyl-2- (trimethylstannyl) -5, 6-dihydro-7H-pyrrolo [3,4-b]A solution of pyridin-7-one (360.0 mg,0.81 mmol) in 1, 4-dioxane (5 mL) was successively added copper iodide (30.4 mg,0.16 mmol), 2, 4-dichloro-5-fluoropyrimidine (203.7 mg,1.22 mmol) and [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (117.1 mg,0.16 mmol) was heated to 100℃under nitrogen and stirred for 2 hours. The reaction was cooled to room temperature and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give a yellow solid (280.0 mg). LC-MS: [ M+H ] ] + :413.2,415.2。
Synthesis of 2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6- (4-methoxybenzyl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (7):
to 2- (2-chloro-5-fluoropyrimidin-4-yl) -6- (4-methoxybenzyl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b]To a solution of pyridin-7-one (260.0 mg,0.63 mmol) in 1, 4-dioxane (10 mL) was added 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (199.0 mg,0.90 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (40.8 mg,0.066 mmol), tris (dibenzylideneacetone) dipalladium (45.0 mg,0.049 mmol) and cesium carbonate (617.5 mg,1.90 mmol) in this order, and the mixture was heated to 100℃and stirred for 16 hours. The reaction solution was cooled to room temperature and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (0-20% methanol/dichloromethane) to give a yellow solid (70.0 mg). LC-MS: [ M+H ]] + :598.3。
Synthesis of 2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b ] pyridin-7-one (DP-01-246):
to 2- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -6- (4-methoxybenzyl) -5, 5-dimethyl-5, 6-dihydro-7H-pyrrolo [3,4-b]To a solution of pyridin-7-one (60.0 mg,0.10 mmol) in trifluoroacetic acid (2 mL) was added trifluoromethanesulfonic acid (1 mL) and heated to 60℃and stirred for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo, and the crude product was purified by prep. to give a yellow solid (5.1 mg). 1 HNMR(400MHz,DMSO)δ9.22(s,1H),8.72(d,J＝3.1Hz,1H),8.36(d,J＝8.0Hz,1H),8.27–8.20(m,2H),8.17(s,1H),7.94(d,J＝2.8Hz,1H),6.92(d,J＝8.9Hz,1H),6.59(dd,J＝8.9,2.9Hz,1H),3.80(s,3H),3.06–3.02(m,4H),2.47–2.43(m,4H),2.22(s,3H),1.53(s,6H)；LC-MS:[M+H] + :478.3。
EXAMPLE 79 Synthesis of Compounds DP-01-266, DP-01-266A, DP-01-266B
Synthesis of 2- (4-methoxy-3-nitrophenyl) -5-methyl-2, 5-diazabicyclo [2.2.1] heptane (2):
4-bromo-1-methoxy-2-nitrobenzene (300.0 mg,1.29 mmol), 2-methyl-2, 5-diazabicyclo [2.2.1]Heptane dihydrochloride (239.3 mg,1.29 mmol), sodium t-butoxide (621.3 mg,6.47 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (123.3 mg,0.26 mmol) and tris (dibenzylideneacetone) dipalladium (118.4 mg,0.13 mmol) were dissolved in toluene (7 mL) and reacted at 90℃for 3 hours under nitrogen protection. The crude product obtained by spin-drying the reaction solution was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:1) to obtain a yellow solid (290.0 mg). LC-MS: [ M+H ]] + :263.9。
Synthesis of 2-methoxy-5- (5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) aniline (3):
the compound 2- (4-methoxy-3-nitrophenyl) -5-methyl-2, 5-diazabicyclo [2.2.1]Heptane (290.0 mg,1.10 mmol) was dissolved in methanol (10 mL), 10% palladium on carbon (30.0 mg) was added and reacted at room temperature under hydrogen protection for 18 hours. For reaction liquidsThe mixture was filtered through celite, and the filtrate was dried to give a brown solid (250.0 mg). LC-MS: [ M+H ]] + :233.9。
Synthesis of 5' - (5-fluoro-2- ((2-methoxy-5- (5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-266):
2-methoxy-5- (5-methyl-2, 5-diazabicyclo [ 2.2.1)]Heptan-2-yl) aniline (250.0 mg,1.07 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (310.4 mg,1.07 mmol), cesium carbonate (1047.4 mg,3.21 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (133.5 mg,0.21 mmol) and palladium acetate (24.1 mg,0.11 mmol) were dissolved in dioxane (5 mL) and reacted at 100℃for 18 hours under nitrogen protection. The crude product obtained by concentration of the reaction solution was prepared by acidic reverse phase to obtain brown solid (192.9 mg). 1 H NMR(400MHz,MeOD)δ8.55(s,1H),8.53(s,1H),8.43(d,J＝3.6Hz,1H),8.27(d,J＝8.0Hz,1H),8.01(d,J＝2.8Hz,1H),7.36(d,J＝8.0Hz,1H),6.89(d,J＝8.8Hz,1H),6.26(dd,J＝8.8,2.8Hz,1H),4.45(s,1H),4.11(s,1H),3.86(s,3H),3.71(dd,J＝10.8,2.4Hz,1H),3.49(t,J＝12.0Hz,2H),3.17(d,J＝9.6Hz,1H),2.79(s,3H),2.23(q,J＝9.6Hz,2H),1.67–1.57(m,4H)；LC-MS:[M+H] + :487.5。
The racemate DP-01-266 was purified by SFC separation (column: chiralPak IH,150*20mm I.D, 5 μm; mobile phase: A phase: CO) 2 The method comprises the steps of carrying out a first treatment on the surface of the Phase B MEOH+0.1% NH 3 H 2 O; gradient: b%:50%;20min,3 h) to give white solids DP-01-266A and white solids DP-01-266B. DP-01-266A:
chiral analysis conditions: chromatographic column: CHIRALPAK IH 5 μm 4.6 x 100mm, mobile phase: phase A: CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH; gradient: b%:50.0%, retention time 6.736 min, ee% = 100%.
1 H NMR(400MHz,MeOD)δ8.53(s,1H),8.46(d,J＝3.6Hz,1H),8.31(d,J＝8.0Hz,1H),8.01(d,J＝2.8Hz,1H),7.38(d,J＝8.0Hz,1H),6.88(d,J＝8.8Hz,1H),6.24(dd,J＝8.8,2.8Hz,1H),4.25(s,1H),3.86(s,3H),3.56–3.51(m,2H),3.33(s,1H),2.84–2.81(m,2H),2.38(s,3H),1.98(q,J＝8.4Hz,2H),1.66–1.59(m,4H)；LC-MS:[M+H] + :487.5。
DP-01-266B：
Chiral analysis conditions: chromatographic column: CHIRALPAK IH 5 μm 4.6 x 100mm, mobile phase: phase A CO 2 The method comprises the steps of carrying out a first treatment on the surface of the And B phase: meOH; gradient: b%:50.0%, retention time 20.346 min, ee% = 100%.
1 H NMR(400MHz,MeOD)δ8.54(s,1H),8.47(d,J＝3.6Hz,1H),8.32(d,J＝8.0Hz,1H),8.02(d,J＝2.8Hz,1H),7.39(d,J＝8.0Hz,1H),6.88(d,J＝8.8Hz,1H),6.24(dd,J＝8.8,2.8Hz,1H),4.25(s,1H),3.86(s,3H),3.55–3.50(m,2H),3.33(s,1H),2.85–2.78(m,2H),2.37(s,3H),1.98(q,J＝8.0Hz,2H),1.68–1.58(m,4H)；LC-MS:[M+H] + :487.5。
EXAMPLE 80 Synthesis of Compounds DP-01-270
Synthesis of 7- (4-methoxy-3-nitrophenyl) -9-methyl-3-oxo-7, 9-diazabicyclo [3.3.1] nonane (2):
4-bromo-1-methoxy-2-nitrobenzene (118.7 mg,0.51 mmol), 9-methyl-3-oxa-7, 9-diazabicyclo [3.3.1] under nitrogen]Nonane dihydrochloride (100.0 mg,0.46 mmol), tris (dibenzylideneacetone) dipalladium (42.6 mg,0.047 mmol), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (44.4 mg,0.093 mmol) and cesium carbonate (606.2 mg,1.86 mmol) were dissolved in toluene (3 mL) and reacted with stirring at 90℃for 5 hours. LCMS detected complete reaction. The reaction mixture was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography on silica gel (dichloromethane: ethyl acetate=20:1) to give a yellow oil (110.0 mg). LC-MS: [ M+H ]] + :294.2。
Synthesis of 2-methoxy-5- (9-methyl-3-oxo-7, 9-diazabicyclo [3.3.1] non-7-yl) aniline (3):
7- (4-methoxy-3-nitrophenyl) -9-methyl-3-oxo-7, 9-diazabicyclo [3.3.1]Nonane (110.0 mg,0.38 mmol) was dissolved in methanol (2 mL) and tetrahydrofuran (1 mL), and 10% palladium on carbon (39.9 mg) The reaction system was added and the reaction was carried out at room temperature for 18 hours under normal pressure after three times of hydrogen substitution. LCMS detected complete reaction. The reaction solution was concentrated by filtration to give a yellow oil (80.0 mg). LC-MS: [ M+H ] ] + :264.3。
Synthesis of 5' - (5-fluoro-2- ((2-methoxy-5- (9-methyl-3-oxo-7, 9-diazabicyclo [3.3.1] non-7-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline ] -3' -one (DP-01-270):
2-methoxy-5- (9-methyl-3-oxo-7, 9-diazabicyclo [ 3.3.1)]Non-7-yl) aniline (80.0 mg,0.30 mmol), 5'- (2-chloro-5-fluoropyrimidin-4-yl) spiro [ cyclopropane-1, 1' -isoindoline]3' -Ketone (88.0 mg,0.30 mmol), tris (dibenzylideneacetone) dipalladium (27.8 mg,0.03 mmol), 1' -binaphthyl-2, 2' -bisdiphenylphosphine (18.9 mg,0.03 mmol) and cesium carbonate (296.9 mg,0.91 mmol) were placed in 1, 4-dioxane (2 mL), nitrogen was replaced three times, and the reaction solution was stirred at 90℃for 5 hours. LCMS detected completion of the reaction, and the reaction mixture was cooled to room temperature, diluted with water (10 mL) and extracted with ethyl acetate (20 ml×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. Purification by column chromatography (dichloromethane: methanol=20:1) followed by purification yielded a yellow solid (2.5 mg). 1 H NMR(400MHz,DMSO)δ8.90(s,1H),8.65(d,J＝3.2Hz,1H),8.30(s,1H),8.23(d,J＝8.0Hz,1H),8.18(s,1H),7.90(d,J＝2.4Hz,1H),7.47(d,J＝8.4Hz,1H),6.95(d,J＝8.8Hz,1H),6.48(dd,J＝8.8,2.8Hz,1H),3.83-3.80(m,5H),3.71(d,J＝10.4Hz,2H),3.36–3.34(m,1H),3.30–3.29(m,1H),3.23(dd,J＝11.6,2.8Hz,2H),2.78(s,2H),2.46(s,3H),1.54(d,J＝2.9Hz,4H)；LCMS:[M+H] + :517.3。
EXAMPLE 81 Synthesis of Compound PR-01-073
Synthesis of methyl 2-cyano-6-methylbenzoate (2):
the compound methyl 2-bromo-6-methylbenzoate (3.0 g,13.10 mmol), cuprous cyanide (1.4 g,15.63 mmol) and cuprous iodide (0.25 g,1.31 mmol) were suspended in N, N-dimethylformamide (20 mL), stirred at 130℃under nitrogen for 18 hours, and cooled to room Temperature. The crude product was chromatographed on a column of silica gel (ethyl acetate: petroleum ether=1:10) to give a white solid (1.1 g). 1 H NMR(400MHz,CDCl 3 )δ7.58(dd,J＝7.2,1.2Hz,1H),7.51–7.40(m,2H),4.01(s,3H),2.48(s,3H)。
Synthesis of 7-methylisoindolin-1-one (3):
the compound methyl 2-cyano-6-methylbenzoate (1.0 g,5.71 mmol) was dissolved in ethanol (80 mL), and Raney nickel alloy (100.0 mg) was added. The reaction was carried out at room temperature for 18 hours under 150psi hydrogen pressure. The reaction solution was filtered through celite, and the filtrate was dried by spinning to give a white solid (800.0 mg). The product was used directly in the next reaction. LCMS [ M+H ]] + :148.3。
Synthesis of 6-iodo-7-methylisoindolin-1-one (4):
in an ice-water bath, 7-methylisoindolin-1-one (800.0 mg,5.44 mmol) was dissolved in concentrated sulfuric acid (8 mL) and stirred for 0.5 hours, N-iodosuccinimide (2.29 g,10.18 mmol) was added to the reaction system, and after stirring the reaction in an ice-water bath for 2 hours, the reaction solution was poured into ice water and quenched. Extraction with ethyl acetate (10 ml×3), washing the organic phase with saturated aqueous sodium sulfite (15 mL) and water (15 mL), drying over anhydrous sodium sulfate, concentrating the crude product, purifying by column chromatography to give (petroleum ether: ethyl acetate=1:1) as a white solid compound (300.0 mg). 1 HNMR(400MHz,DMSO-d6)δ8.57(s,1H),7.98(d,J＝8.0Hz,1H),7.18(d,J＝8.0Hz,1H),4.24(s,2H),2.70(s,3H)。
Synthesis of 7-methyl-6-boronic acid pinacol ester isoindolin-1-one (5):
The compound 6-iodo-7-methylisoindolin-1-one (100.0 mg,0.37 mmol), pinacol borane (100.0 mg,0.78 mmol), triethylamine (150.0 mg,1.48 mmol), 2- (dicyclohexylphosphino) biphenyl (26.0 mg,0.074 mmol) and palladium acetate (5.0 mg,0.022 mmol) were suspended in dioxane (2 mL) and reacted at 80℃for 1 hour under nitrogen. The reaction was cooled, diluted with water (8 mL), and extracted with ethyl acetate (8 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, and spin-dried to give a crude product (70.0 mg, purity: 40%) which was directly used for the next reaction. LCMS [ M+H ]] + :274.3。
Synthesis of 7-methyl-6- (2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) isoindolin-1-one (PR-01-073):
the compound 7-methyl-6-boronic acid pinacol ester isoindolin-1-one (70.0 mg,0.10mmol, purity: 40%), 4-chloro-N- (5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) pyrimidin-2-amine (50.0 mg,0.13 mmol), potassium carbonate (45.5 mg,0.33 mmol) and [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride dichloromethane complex (8.1 mg,0.01 mmol) was dissolved in dioxane (5 mL) and water (1 mL), reacted for 3 hours at 100℃under nitrogen protection and concentrated directly to dryness. Purification by acid preparation gave a white solid (24.7 mg). 1 H NMR(400MHz,MeOD)δ8.55(d,J＝5.2Hz,1H),8.21(s,1H),7.68(d,J＝7.6Hz,1H),7.51(d,J＝8.0Hz,1H),7.25(d,J＝9.2Hz,1H),7.09–7.07(m,1H),6.76(dd,J＝9.2,2.8Hz,1H),4.45(s,2H),3.82(d,J＝12.4Hz,2H),3.60–3.54(m,2H),3.27–3.24(m,2H),3.13–3.05(m,2H),2.96(s,3H),2.77(s,3H)；LC-MS:[M+H] + :499.5。
EXAMPLE 82 Synthesis of Compounds DP-01-274
Synthesis of 2' - (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -5' (4'H) -carboxylic acid tert-butyl ester (1):
2' - (2-chloro-5-fluoropyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-5 '(4'H) -carboxylic acid tert-butyl ester (50.0 mg,0.13 mmol), 2-methoxy-5- (4-methylpiperazin-1-yl) aniline (28.0 mg,0.13 mmol), cesium carbonate (123.0 mg,0.38 mmol), tris (dibenzylideneacetone) dipalladium (12.0 mg,0.013 mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (16.0 mg,0.026 mmol) were placed in anhydrous dioxane (1 mL) and reacted at 100 ℃ under nitrogen for 3 hours. LCMS monitored reaction was complete. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane: methanol=10:1) to give a yellow solid (50.0 mg). LC-MS: [ M+H ]] + :581.5。
Synthesis of 2'- (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -4 '(5' H) -one (DP-01-274):
2' - (5-fluoro-2- ((2-methoxy-5- (4-methylpiperazin-1-yl) phenyl) amino) pyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds ]Tert-butyl 5 '(4'H) -carboxylate (50.0 mg,0.086 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.2 mL) was slowly added under ice-bath cooling and the reaction stirred at room temperature for 1 hour. LCMS monitored reaction was complete. The reaction solution was concentrated and purified by ammonium bicarbonate to give a yellow solid (30.4 mg). 1 H NMR(400MHz,DMSO-d6)δ8.71(s,1H),8.65(d,J＝3.2Hz,1H)8.10(s,1H),7.96(d,J＝2.8Hz,1H),7.80(s,1H),6.92(d,J＝8.8Hz,1H),6.58(dd,J＝8.8,2.8Hz,1H),3.81(s,3H),3.10–3.07(m,4H),2.49–2.46(m,4H),2.24(s,3H),1.69–1.67(m,2H),1.56–1.54(m,2H)；LC-MS:[M+H] + :481.2。
EXAMPLE 83 Synthesis of Compound DP-01-275
Synthesis of 2' - (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -5' (4'H) -carboxylic acid tert-butyl ester (1):
2' - (2-chloro-5-fluoropyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2,3-c ]]Pyrrole compounds]-5 '(4'H) -carboxylic acid tert-butyl ester (40.0 mg,0.10 mmol), 5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) aniline (28.0 mg,0.10 mmol), cesium carbonate (99.0 mg,0.30 mmol), tris (dibenzylideneacetone) dipalladium (9.2 mg, 0.010mmol) and 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (13.5 mg,0.022 mmol) were placed in anhydrous dioxane (1 mL) and reacted at 100 ℃ under nitrogen for 3 hours. LCMS monitored reaction was complete. The reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane: methanol=10:1) to give a yellow solid (50.0 mg). LC-MS: [ M+H ]] + :635.5。
Synthesis of 2'- (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) spiro [ cyclopropane-1, 6' -thieno [2,3-c ] pyrrole ] -4 '(5' H) -one (DP-01-275):
2' - (5-fluoro-2- ((5- (4-methylpiperazin-1-yl) -2- (trifluoromethoxy) phenyl) amino) pyrimidin-4-yl) -4' -oxospiro [ cyclopropane-1, 6' -thieno [2, 3-c)]Pyrrole compounds]Tert-butyl 5 '(4'H) -carboxylate (50.0 mg,0.079 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (0.2 mL) was slowly added after cooling in an ice bath, and the reaction was stirred at room temperature for 1 hour. LCMS monitored reaction was complete. The reaction solution was concentrated and purified by ammonium bicarbonate to give a pale yellow solid (28.2 mg). 1 H NMR(400MHz,DMSO-d6)δ9.02(s,1H),8.69(s,1H),8.62(d,J＝3.2Hz,1H),7.78(s,1H),7.53(d,J＝2.8Hz,1H),7.20(d,J＝9.2Hz,1H),6.74(dd,J＝9.2,2.8Hz,1H),3.20–3.18(m,4H),2.49–2.47(m,4H),2.25(s,3H),1.66–1.64(m,2H),1.55–1.53(m,2H)；LC-MS:[M+H] + :535.1。
Example 84
The following compounds were prepared according to the procedure of example 1:
example 85
The following compounds were prepared according to the procedure for example 13:
example 86
The following compounds were prepared according to the procedure for example 7:
example 87
The following compounds were prepared according to the procedure for example 12:
example 88
The following compounds were prepared according to method III:
example 89
The following compounds were prepared according to the procedure for example 12:
EXAMPLE 90 test of Compounds for PLK1 inhibitory Activity
The test compounds were diluted 3-fold in a 384-well dilution plate with DMSO, and the diluted compounds were transferred 25nL using ECHO in a 384-well reaction plate to ensure a DMSO content of 0.5%. Transfer 2.5. Mu.L of 2 XPLK 1 enzyme solution to 384 well reaction plates, centrifuge at 1000rpm for 1min, incubate at 25℃for 10min. Transfer of 2.5. Mu.L of 2 XATP &PLK tide mixed solution was centrifuged at 1000rpm for 1min in 384 well reaction plates and incubated at 25℃for 60min. Transfer 4. Mu.L ADP-Glo Reagent to 384 well reaction plates, centrifuge at 1000rpm for 1min, incubate at 25℃for 40min. Transfer 8. Mu.L ADP-Glo Detection reagent to 384 well reaction plates, centrifuge at 1000rpm for 1min, incubate at 25℃for 40min. The RLU (Relative luminescence unit) signal was read using a BMG microplate reader. The signal intensity was used to characterize the extent of kinase activity. Calculating positive drug IC by utilizing GraphPad software nonlinear fitting formula (1) 50 ：
Y＝Bottom + (Top-Bottom)/(1+10^((LogIC50-X) ×HillSlope)) (1)
Wherein X represents the log of the compound concentration and Y represents the inhibition (% inhibition).
The results show that the compounds of the invention have good PLK1 inhibitory activity with an IC50 of less than 100nM, preferably less than 10nM, and that the IC50 of representative compounds are shown in Table 1 below:
TABLE 1 inhibitory Activity of representative Compounds against PLK1
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A compound represented by the following formula (I),
or a pharmaceutically acceptable salt, deuterated thereof, wherein:
E is independently selected from C, N, O;
t, M, Q is independently selected from N, CR3;
x, Y, Z is independently selected from N, CR;
ring a is selected from phenyl, containing 1-3 5-11 membered heteroaryl groups independently selected from N, O, S;
the B ring is connected with the ring carbon atom or the ring nitrogen atom of the A ring;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, optionally one or more of which are independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from one or more of F, cl, br, OH, CN, -NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-11 membered heteroaryl independently selected from N, O, S;
R4 is selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r6 is selected fromNR8R9、NR8C(O)R9、C(O)NR8R9、C 1-4 Alkyl, piperazinyl, bridged piperazinyl, spiropiperazinyl, said alkyl, piperazinyl, bridged piperazinyl, spiropiperazinyl optionally substituted with one or more R7;
or R6 together with the ring C atom, ring Y atom, and the bond between the ring C atom and ring Y atom to which it is attached form a ring containing 1 to 3 independent groupsA 5-7 membered heterocycloalkyl group optionally selected from N, O, S, which ring is optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r7 is independently selected from H, F, cl, br, OH, CN, NR R9, = O, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, said alkyl, alkoxy, cycloalkyl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, said alkyl, alkenyl, alkynyl, alkoxy optionally being substituted with one or more groups selected from F, cl, br, OH, CN, NH 2 Is substituted by a group of (2);
when R8 and R9 are attached to the same atom, R8 and R9 together with the atom to which they are attached may form a 3-7 membered heterocycloalkyl containing 1-3 members independently selected from N, O, S, optionally substituted with one or more members independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
m is 0, 1, 2 or 3.
w is 0, 1, 2.
2. The compound of claim 1, having the structure of formula (IA):
Wherein:
t, M, Q is independently selected from N, CR3;
x, Y, Z is independently selected from N, CR;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, which cycloalkyl group may optionally be independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkyneOptionally one or more of the radicals, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl are independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being independently selected from one or more of F, cl, br, OH, CN, -NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S;
r4 is selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
r7 is independently selected from H, F, cl, br, OH, CN, NR R9, = O, C 1-4 Alkyl, C 3-7 Cycloalkyl groups, said alkyl and cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, said alkyl, alkenyl, alkynyl, alkoxy optionally being substituted with one or more groups selected from F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
when R8 and R9 are attached to the same atom, R8 and R9 together with the atom to which they are attached may form a 3-7 membered heterocycloalkyl containing 1-3 members independently selected from N, O, S, optionally substituted with one or more members independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy groups optionally substituted with one or more groups independently selected from F, cl, br, OH, CN, NH 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 The group of the alkoxy group is substituted;
m is 0, 1, 2 or 3;
n is 0, 1, 2, 3, 4, 5, 6, 7 or 8.
Preferably:
t is N, M, Q are independently selected from CR3;
x, Y, Z is independently selected from CR5;
r1 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl groups, optionally one or more of which are independently selected fromF、Cl、Br、OH、CN、NH 2 Is substituted by a group of (2);
when R1 is independently C 1-4 When alkyl, both R1's together with the C to which they are attached may form a 3-7 membered cycloalkyl group, which cycloalkyl group may optionally be independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
r2 is independently selected from H, F, cl, br, OH, CN, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups are substituted;
r3 is independently selected from H, F, cl, br, OH, CN, NO 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl, containing 1-3-7 membered heterocycloalkyl independently selected from N, O, S, containing 1-3 5-6 membered heteroaryl independently selected from N, O, S, -C (O) R8, -C (O) NR8R9, -S (O) 2 R8、-S(O) 2 NR8R9、-NR8R9, -NR8C (O) R9, -P (O) R8R9, said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycloalkyl, heteroaryl optionally being selected independently from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, C 1-4 Alkoxy, C 3-7 Cycloalkyl groups are substituted;
r4 is selected from C 1-4 Alkoxy, said alkoxy optionally being substituted with oneSubstituted with one or more groups independently selected from F, cl, br;
r5 is independently selected from H, F, cl, br, OH, CN, NH 2 、C 1-4 An alkyl group, optionally one or more independently selected from F, cl, br, OH, CN, NH 2 Is substituted by a group of (2);
r7 is independently selected from H, F, cl, br, OH, CN, NH 2 、＝O、C 1-4 Alkyl, C 3-7 Cycloalkyl groups, said alkyl and cycloalkyl groups optionally being independently selected from one or more of F, cl, br, OH, CN, NH 2 、C 1-4 The group of the alkyl group is substituted;
r8 and R9 are independently selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy groups, said alkyl groups, alkoxy groups optionally being independently selected from F, cl, br, OH, CN, NH by one or more 2 、C 1-4 The group of the alkyl group is substituted;
m is 0, 1, 2 or 3;
n is 0, 1, 2, 3 or 4.
3. A compound according to claim 1 or claim 2, having the structure (IC) as shown in the following formula (IC):
wherein,
x, Y is independently selected from N, CR;
r1 is independently selected from H, OH, = O, C 1-4 An alkyl group;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r2 is independently selected from H, C 1-4 Alkyl, -S (O) 2 R8、-P (O) R8R9, said alkyl groups optionally being independently selected from one or more of OH, C 1-4 The group of the alkoxy group is substituted;
r3 is selected from H, F, cl, br, CN, C 1-4 An alkyl group;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, cyclopropenyl optionally substituted with one or more groups independently selected from F, cl, br, CN, OH;
r5 is independently selected from H, CN;
r7 is independently selected from H, C 1-4 An alkyl group optionally substituted with one or more groups independently selected from F, cl, br, OH, CN;
r8 and R9 are independently selected from NH 2 、C 1-4 An alkyl group.
Preferably, the structure is as shown in the following formula (ID):
Wherein,
r1 is independently selected from H, =o, methyl;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r3 is selected from H, F, cl, br, CN and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, cyclopropenyl optionally substituted with one or more groups independently selected from F, cl, br, CN, OH.
More preferably, the structure is represented by the following formula (IB):
wherein:
r1 is independently selected from H, methyl;
r3 is selected from H, F, cl, br and methyl.
Alternatively, more preferably, the structure is as shown in the following formula (IF):
wherein,
r3 is selected from H, F, cl, br and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, said alkyl, alkoxy optionally substituted with one or more groups independently selected from F, cl, br; preferably methoxy, trifluoromethoxy;
r6 is selected from
4. The compound of claim 1, wherein the structure is represented by the following formula (IE):
wherein,
r1 is independently selected from H, methyl;
when R1 are both methyl, both R1 together with the C to which they are attached may form cyclopropane group;
r3 is selected from H, F, cl, br and methyl;
r4 is selected from C 1-4 Alkyl, C 1-4 Alkoxy, said alkyl, alkoxy optionally substituted with one or more groups independently selected from F, cl, br.
5. The compound of claim 1, wherein the compound is selected from the group consisting of:
6. a pharmaceutical composition comprising a compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.
7. A method of inhibiting PLK1, wherein a compound according to any one of claims 1-5 is used.
8. A method of preventing and/or treating a PLK 1-related disorder, wherein a therapeutically and/or prophylactically effective amount of a compound according to any one of claims 1-5 and a pharmaceutical composition according to claim 6 is administered to a subject in need thereof.
Preferably, the PLK 1-related disease is cancer.
More preferably, the cancer is selected from the group consisting of rectal cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, gastric cancer, head and neck cancer, ovarian cancer, uterine cancer, glioma, liver cancer, esophageal cancer, bladder cancer, renal cancer, lymphoma, melanoma, osteosarcoma, leukemia, myelodysplastic syndrome.
9. Use of a compound according to any one of claims 1-5 and a pharmaceutical composition according to claim 6 for the preparation of a PLK1 inhibitor.
10. Use of a compound according to any one of claims 1-5 and a pharmaceutical composition according to claim 6 for the manufacture of a medicament for the prevention and/or treatment of PLK 1-related diseases.
Preferably, the PLK 1-related disease is cancer.
More preferably, the cancer is selected from the group consisting of rectal cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, gastric cancer, head and neck cancer, ovarian cancer, uterine cancer, glioma, liver cancer, esophageal cancer, bladder cancer, renal cancer, lymphoma, melanoma, osteosarcoma, leukemia, myelodysplastic syndrome.
CN202311140853.0A 2022-09-08 2023-09-06 Compounds as PLK1 inhibitors, and preparation method and application thereof Pending CN117658987A (en)

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GB0317841D0 (en) * 2003-07-30 2003-09-03 Cyclacel Ltd Compound
US7521457B2 (en) * 2004-08-20 2009-04-21 Boehringer Ingelheim International Gmbh Pyrimidines as PLK inhibitors
WO2009040399A1 (en) * 2007-09-28 2009-04-02 Nerviano Medical Sciences S.R.L. Substituted pyrrolo-pyrimidine derivatives, process for their preparation and their use as kinase inhibitors
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