CN115232144A - Nitrogen-containing fused ring derivative, pharmaceutical composition, preparation method and application thereof - Google Patents

Nitrogen-containing fused ring derivative, pharmaceutical composition, preparation method and application thereof Download PDF

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CN115232144A
CN115232144A CN202210425594.5A CN202210425594A CN115232144A CN 115232144 A CN115232144 A CN 115232144A CN 202210425594 A CN202210425594 A CN 202210425594A CN 115232144 A CN115232144 A CN 115232144A
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CN115232144B (en
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宋云龙
沈光远
刘鹏
李曼华
金磊
王国成
苗新园
黄悦
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Changchun Genescience Pharmaceutical Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/12—Drugs for genital or sexual disorders; Contraceptives for climacteric disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/16—Masculine contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/18—Feminine contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/44—Iso-indoles; Hydrogenated iso-indoles
    • C07D209/48—Iso-indoles; Hydrogenated iso-indoles with oxygen atoms in positions 1 and 3, e.g. phthalimide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Abstract

The invention provides a nitrogen condensed ring derivative shown as a formula I, a pharmaceutical composition, and a preparation method and application thereof. The compound has good GnRH receptor antagonism and longer half-life, and can be used for treating or preventing gonad hormone-related diseases and symptoms and preparing medicaments for the diseases and symptoms.

Description

Nitrogen-containing fused ring derivative, pharmaceutical composition, preparation method and application thereof
The invention claims priority of prior application named as 'nitrogenous fused ring derivatives, pharmaceutical compositions, preparation methods and applications' with patent application number 202110438605.9, which is submitted to the intellectual property office of China at 22.4.2021. The entire disclosure of this prior application is incorporated herein by reference.
Technical Field
The invention belongs to the field of medicines, and particularly relates to a nitrogen-containing fused ring derivative, a pharmaceutical composition, and a preparation method and application thereof.
Background
Gonadotropin-releasing hormone (GnRH), also known as luteinizing hormone-releasing hormone (LH-RH), is a decapeptide hormone (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH) synthesized by the neuroendocrine cells of the hypothalamus 2 ) Are central regulatory factors in the endocrine-reproductive system. Which is transported to the pituitary via the hypothalamic pituitary portal circulation, binds to GnRH receptor cells of the anterior pituitary, and secretion and release of gonadotropins such as Luteinizing Hormone (LH) and Follicle Stimulating Hormone (FSH), regulates the normal development of the ovary and corpus luteum, and plays an important role in the hypothalamic-pituitary-gonadal axis. The GnRH receptor exerts its regulatory role by coupling to G proteins capable of activating the calcium second messenger system of phosphatidylinositol, whereas LH regulates the production of sex steroids and FSH regulates spermatogenesis in males and development of follicles in females.
LH and FSH are released into the circulation and bind to receptors on specific cells of the ovary or testis, stimulating steroid production. In the presence of sex steroids, disease conditions such as endometriosis, uterine fibroids and prostate cancer are exacerbated and therapeutic control by administration of long-acting peptide GnRH receptor agonists and antagonists.
Peptide GnRH receptor antagonists include cetrorelix, ganirelix, etc., which block GnRH from binding to its receptor primarily by rapidly competing with GnRH receptors, and block formation of dimer complexes and signal transmission, thereby inhibiting the release of LH and FSH. However, peptide compounds have many problems to be solved including oral absorbability, dosage form, dose volume, drug stability, sustained action, and metabolic stability. The main reason why the treatment with the small-molecule GnRH receptor antagonist is superior to the existing peptide treatment method is that the small-molecule GnRH receptor antagonist can be directly orally taken, and is convenient and quick. Research proves that the small molecule antagonist has obvious curative effect on hormone-dependent diseases such as endometriosis, precocious puberty, prostatic cancer and the like.
In view of the limitations of peptidic GnRH receptor antagonists, several non-peptidic GnRH receptor antagonists have been proposed and put into development, clinical trials and marketing, for example Elagolix developed by AbbVie corporation has been approved for the treatment of endometriosis, while uterine fibroid indications have been declared on the market; relugolix developed by Takada corporation has been approved for the treatment of uterine fibroids, and indications for both endometriosis and prostate cancer have entered the third clinical stage; linzagolix, developed by the cooperation of ObsEva and Kissei corporation, was in the third stage of clinical trials for both endometriosis and uterine fibroids.
Although a great deal of significant research has been conducted in this field, there is a continuing need to develop more effective small molecule GnRH receptor antagonists, and the present invention provides a compound having a novel structure as a GnRH receptor antagonist and finds that a compound having such a structure has excellent activity for effectively treating endocrine-reproductive system diseases.
Disclosure of Invention
In order to improve the technical problems, the invention provides a compound shown as a formula I, a racemate, a stereoisomer, a tautomer, an isotopic marker, a solvate, a polymorph, a pharmaceutically acceptable salt or a prodrug compound thereof:
Figure BDA0003608368340000021
Wherein Y is selected from the following groups, unsubstituted or substituted by X and/or-O-J: c 6-20 An aromatic or 5-20 membered heteroaromatic ring; preferably, A and-E-G are in the meta position, X and-O-J are in the meta position, A and X are in the ortho position, -E-G and-O-J are in the ortho position;
a is selected from unsubstituted or optionally substituted by one, two or more R 1 Substituted by
Figure BDA0003608368340000022
Or
Figure BDA0003608368340000023
B is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted 5-20 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-20 membered heterocyclic ring or 5-20 membered heteroaromatic ring;
e is selected from O, C (O), NH, NC 1-40 Alkyl, N (C) 3-20 Cycloalkyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、NHC(O)、OCH 2 、O(CH 2 ) 2 、NHCH 2 Or N (C) 1-40 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-40 Alkyl radical, C 6-20 Aryl, 5-20 membered heterocyclyl or 5-20 membered heteroaryl;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-40 Alkyl radical, C 3-20 Cycloalkyl radical, C 6-20 Aryl, 5-20 membered heteroaryl, or J-O-fused to the Y to which it is attached, preferably to the phenyl ring to which it is attached, to form a 5-20 membered heteroaryl ring;
x is selected from hydrogen or halogen;
Figure BDA0003608368340000024
is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyloxy radical, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, -NH 2 、-COOH、-COOM、-NH-C(O)-NH 2 、-C(O)C 1-40 Alkyl, -C (O) OC 1-40 Alkyl, -C (O) C 1-40 Hydroxyalkyl, -S (O) 2 -NHC 1-40 Alkyl, -S (O) 2 C 6-20 Aryl, -S (O) 2 -5-20 membered heteroaryl, NH-C (O) -NH-OC 1-40 Alkyl or NH 2 -C (O) -; wherein M is selected from cations;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 、COOH、NH-C(O)-NH 2 Or NH-C (O) -NH-OC 1-40 An alkyl group;
each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-40 Alkyl radical, C 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl group, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkanoyloxy groupBase, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 。
According to an embodiment of the invention, Y is selected from the following groups, unsubstituted or substituted by X and/or-O-J: a benzene ring or a pyridine ring;
b is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted 5-14 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-14 membered heterocyclic ring or 5-14 membered heteroaromatic ring;
e is selected from O, C (O), NH, NC 1-8 Alkyl, N (C) 3-8 Cycloalkyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 、NHC(O)、NHCH 2 Or N (C) 1-8 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl radical, C 6-14 Aryl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure BDA0003608368340000031
And B is selected from a thiophene ring, G is selected from a 9-14 membered heteroaryl or a 9-14 membered heterocyclyl, said heteroaryl or heterocyclyl preferably being a fused ring group;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl radical, C 3-8 Cycloalkyl, C 6-14 Aryl, 5-14 membered heteroaryl, or J-O-is attached to the C at the ortho position of the phenyl ring to which it is attached to form a 5-14 membered heteroaromatic ring which is attached to the phenyl ring in juxtaposition;
x is selected from hydrogen or halogen;
Figure BDA0003608368340000032
is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 、COOH、-COOM、NH-C(O)-NH 2 、-C(O)C 1-8 Alkyl, -C (O) OC 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl, NH-C (O) -NH-OC 1-8 Alkyl or NH 2 -C (O) -; wherein M is selected from monovalent cations;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 、COOH、NH-C(O)-NH 2 Or NH-C (O) -NH-OC 1-8 An alkyl group.
Each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-8 Alkyl radical, C 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyloxy radical, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 。
According to an embodiment of the invention, said B is selected from unsubstituted or optionally substituted by one, two or more R 2 Substituted thiophene, furan, imidazole, pyrazole rings;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 Substituted thiophene, furan, imidazole, pyrazole, pyrrole, tetrahydropyrrole rings;
E is selected from O, C (O), NH, NCH 3 N (-cyclopropyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 、NHC(O)、NHCH 2 Or N (CH) 3 )CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure BDA0003608368340000033
And B is selected from the thiophene ring, G is selected from unsubstituted or optionally substituted with one, two or more R 4 Substituted C 9-14 An aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl group which may be a fused ring group;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl radical, C 3-8 A cycloalkyl group or J-O-is linked to C at the ortho position in the benzene ring to which it is linked, to form an oxazole ring, thiazole ring, isoxazole ring, pyrrole ring, furan ring, imidazole ring, pyridine ring, or pyrazine ring which is in parallel with the benzene ring;preferably, J is selected from H, methyl, cyclopropylmethyl, n-butyl, methoxyethyl, cyclopentyl;
x is selected from hydrogen, fluorine, chlorine or bromine;
Figure BDA0003608368340000041
is a linking site;
each R 1 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOM, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C (O) C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, ph-NH-C (O) -NH-OC 1-8 Alkyl radical, C 1-8 alkyl-NH-C (O) -, di (C) 1-8 Alkyl) N-C 1-8 Alkyl, hydroxy C 1-8 An alkyl group; m is selected from monovalent cations, e.g. Na + 、K + Or
Figure BDA0003608368340000042
Each R 2 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOM, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C (O) C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, ph-NH-C (O) -NH-OC 1-8 Alkyl radical, C 1-8 alkyl-NH-C (O) -, di (C) 1-8 Alkyl) N-C 1-8 Alkyl, hydroxy C 1-8 An alkyl group; m is selected from monovalent cations, e.g. Na + 、K + Or
Figure BDA0003608368340000043
Each R 3 Are identical or different and are each independently selected from hydrogen, oxo (= O), C 1-8 Alkyl, COOH, COOC 1-8 An alkyl group;
each R 4 Identical or different, independently of one another, from hydrogen, halogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl or C 1-8 An alkyloxy group;
each R 5 Identical or different, independently of one another, from hydrogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl, C 1-8 An alkyloxy group;
each R a Identical or different, independently of one another, from hydrogen, halogen, C 1-8 Alkyl radical, C 3-8 Cycloalkyl radical, C 1-8 An alkyloxy group.
According to an embodiment of the invention, the structure of the compound of formula I is as shown in formula I':
Figure BDA0003608368340000044
Wherein A, E, G, J, X has the definition described above.
According to an embodiment of the invention, the compound of formula I has the structure shown in formula I-1 or I-2:
Figure BDA0003608368340000045
wherein B, D, E, G, J, X, R 1 Having the definition as described above, n is selected from an integer from 1 to 4.
According to an embodiment of the invention, the compound of formula I has the structure shown in formula I-3, formula I-4, formula I-5, formula I-6, formula I-7 or formula I-8:
Figure BDA0003608368340000046
Figure BDA0003608368340000051
of these, E, G, J, X, R 1 Having the definitions as described above.
According to an embodiment of the invention, the compound of formula I has the structure shown in formula II:
Figure BDA0003608368340000052
wherein the content of the first and second substances,
e is selected from O, C (O), NH, NCH 3 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 NHC (O), N (-cyclopropyl) CH 2 、NHCH 2 Or N (CH) 3 )CH 2 ；
G has the meaning indicated above, preferably unsubstituted or optionally substituted by one, two or more R 4 Substituted C 9-14 An aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl group, said aryl, heteroaryl or heterocyclyl group being a group of a fused ring;
R 4 having the definitions as described above.
According to an embodiment of the invention, the compound of formula I has the structure shown in formula II:
Figure BDA0003608368340000053
wherein G has the meaning as indicated above, preferably unsubstituted or optionally substituted by one, two or more R 4 Substituted C 9-14 An aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl group, said aryl, heteroaryl or heterocyclyl group being a group of a fused ring;
R 4 Having the definitions as described above.
In a preferred embodiment of the present invention, G is selected from the group consisting of:
Figure BDA0003608368340000054
Figure BDA0003608368340000061
Figure BDA0003608368340000071
according to an embodiment of the invention, the structure of the compound of formula I is as follows:
Figure BDA0003608368340000072
Figure BDA0003608368340000081
Figure BDA0003608368340000091
Figure BDA0003608368340000101
Figure BDA0003608368340000111
Figure BDA0003608368340000121
Figure BDA0003608368340000131
the present invention also provides a process for the preparation of compounds of formula I, comprising the following scheme 1 or scheme 2:
scheme 1: reacting the compound 1 under the action of alkali to obtain a compound shown as a formula I-1;
Figure BDA0003608368340000132
wherein B, E, G, J, X, R 1 And n has the meaning indicated above, R is C 1-8 An alkyl group;
according to an embodiment of the present invention, the base in scheme 1 may be an inorganic base, preferably an alkali metal hydroxide, such as lithium hydroxide, lithium hydroxide monohydrate, sodium hydroxide, potassium hydroxide;
scheme 2: reacting the compound 2 with the compound 3 to obtain a compound shown as a formula I-2;
Figure BDA0003608368340000141
wherein D, E, G, J, X, R 1 And n has the definition set out above; y being a reactive group capable of causing, in a reaction, compound 3 to be substituted or coupled to the D ring of compound 2, e.g. halogen, NH 2 、B(OH) 2 ；
According to an embodiment of the present invention, the reaction in scheme 2 may be carried out in the presence of a base, such as an organic base, which may be at least one of triethylamine, diisopropylethylamine, pyridine, DMAP, DBU; and an organometallic compound such as at least one of lithium diisopropylamide, methyllithium, and butyllithium.
According to an embodiment of the present invention, the reaction described in scheme 1 or scheme 2 may be carried out in the presence of a solvent such as an organic solvent or a mixed solvent of an organic solvent and water. For example, the organic solvent may be selected from at least one of the following: alcohols such as methanol, ethanol, isopropanol, n-butanol; ethers such as ethyl propyl ether, n-butyl ether, anisole, phenetole, cyclohexyl methyl ether, dimethyl ether, diethyl ether, dimethyl glycol, diphenyl ether, propyl ether, isopropyl ether, isobutyl ether, isopentyl ether, ethylene glycol dimethyl ether, isopropylethyl ether, methyl tert-butyl ether, tetrahydrofuran, methyl tetrahydrofuran, dioxane, dichlorodiethyl ether, and polyethers of ethylene oxide and/or propylene oxide; aliphatic, cycloaliphatic or aromatic hydrocarbons, such as pentane, hexane, heptane, octane, nonane, and hydrocarbons which may be substituted by fluorine and/or chlorine atoms, such as dichloromethane, trichloromethane, carbon tetrachloride, fluorobenzene, chlorobenzene or dichlorobenzene; cyclohexane, methylcyclohexane, petroleum ether, acetone, octane, benzene, toluene, chlorobenzene, bromobenzene and xylene; esters such as methyl acetate, ethyl acetate, butyl acetate, isobutyl acetate and dimethyl carbonate, dibutyl carbonate or ethylene carbonate.
The invention also provides a pharmaceutical composition, which comprises at least one of a therapeutically effective amount of compound shown in formula (I), racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.
According to an embodiment of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.
According to an embodiment of the invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.
The present invention also provides a method for treating a sex hormone related disease, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one compound of formula (I), its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof.
The present invention also provides a method for treating a gonadal hormone related disorder comprising administering to a subject a prophylactically or therapeutically effective amount of the above pharmaceutical composition.
The sex-hormone related diseases include sex-hormone dependent cancer, bone metastasis of sex-hormone dependent cancer, prostatic hypertrophy, hysteromyoma, endometriosis, uterine fibroids, precocious puberty, amenorrhea, premenstrual syndrome, dysmenorrhea, multiple-atrial ovarian syndrome, polycystic ovarian syndrome, acne, alopecia, alzheimer's disease, infertility, irritable bowel syndrome, hormone-independent and LH-RH sensitive benign or malignant tumors or flushing; the sex hormone dependent cancer is selected from prostate cancer, uterine tumor, breast cancer, and pituitary cancer.
In some embodiments, the patient comprises a mammal, preferably a human.
The invention also provides at least one of a compound shown as a formula (I), racemate, stereoisomer, tautomer, isotopic marker, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, or a pharmaceutical composition for preventing or treating diseases related to the gonadal hormone.
The invention also provides application of at least one of the compound shown in the formula (I), racemate, stereoisomer, tautomer, isotopic marker, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof in preparing medicines.
According to an embodiment of the present invention, the use may be a use in the manufacture of a medicament for the prevention or treatment of a sex hormone related disease, such as a use in the manufacture of a GnRH receptor antagonist.
According to an embodiment of the present invention, the use may be in the manufacture of a medicament for the treatment or prevention of sex hormone dependent cancer, bone metastasis of sex hormone dependent cancer, prostatic hypertrophy, uterine myoma, endometriosis, uterine fibroids, precocious puberty, amenorrhea, premenstrual syndrome, dysmenorrhoea, multi-atrial ovarian syndrome, polycystic ovarian syndrome, acne, alopecia, alzheimer's disease, infertility, irritable bowel syndrome, hormone-independent and LH-RH sensitive benign or malignant tumors, or diseases of flushing; such as the use in the preparation of reproductive regulators, contraceptives, ovulation inducers; or as a medicament for the preparation of a medicament for the prevention of postoperative recurrence of a sex hormone dependent cancer, wherein the sex hormone dependent cancer is selected from the group consisting of prostate cancer, uterine tumors, breast cancer, pituitary cancer.
The medicament can be used for diseases related to gonadal hormone.
The effective dose of the active compound can vary widely and is generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of the compound actually administered will generally be determined by a physician, in the light of the relevant circumstances, and will include the condition to be treated, the chosen route of administration, the actual compound administered; age, weight and response of the individual patient; severity of patient symptoms, etc.
The amount of compound or composition administered to a patient is not fixed and depends on the drug administered, the purpose of the administration such as prevention or treatment; the condition of the patient, the mode of administration, etc. In therapeutic applications, compositions can be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dosage will depend on the disease state being treated and the judgment of the attending clinician, which will depend on factors such as the severity of the disease, the age, weight and general condition of the patient.
The composition administered to the patient may be in the form of a pharmaceutical composition as described above. These compositions may be sterilized by conventional sterilization techniques or may be sterilized by filtration.
Therapeutic dosages of the compounds of the invention may be determined, for example, by: the particular use of the treatment, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the invention in the pharmaceutical composition may not be fixed and will depend on a variety of factors including the dosage, chemical properties (e.g., hydrophobicity), and the route of administration. The compounds of the present invention can be provided, for example, by containing about 0.1 to 10% w/v of the physiological buffered aqueous solution of the compound for parenteral administration. Some typical dosage ranges are from about 1. Mu.g/kg to about 1g/kg body weight/day. In certain embodiments, the dosage range is from about 0.01mg/kg to about 100mg/kg of body weight per day. The dosage will likely depend on such variables as the type and extent of progression of the disease or disorder, the general health status of the particular patient, the relative biological efficacy of the selected compound, the excipient formulation and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.
Advantageous effects
The compound provided by the invention has good GnRH receptor antagonism, and can be used for treating or preventing gonad hormone-related diseases and symptoms and preparing medicaments for the diseases and symptoms. Also, the half-life of the compounds is longer.
Definition and description of terms
Unless otherwise indicated, the definitions of radicals and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and definitions of groups and structures of compounds after combination are to be understood as being within the scope of the present description and/or claims.
Unless otherwise indicated, the numerical ranges set forth in the specification and claims are equivalent to at least each specific integer recited therein. For example, a numerical range of "1-40" is equivalent to reciting each of the integer values in the numerical range of "1-10," i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and each of the integer values in the numerical range of "11-40," i.e., 11, 12, 13, 14, 15, 35, 36, 37, 38, 39, 40. Further, when certain numerical ranges are defined as "numbers," it should be understood that the two endpoints of the range, each integer within the range, and each decimal within the range are recited. For example, "a number of 0 to 10" should be understood to not only recite each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, but also to recite at least the sum of each integer with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.
It should be understood that in describing 1,2 or more herein, "more" shall mean an integer greater than 2, e.g., greater than or equal to 3, e.g., 3, 4, 5, 6, 7, 8, 9, or 10.
The term "optional" (or "optionally", "optionally") means substituted with zero, one or more substituents, e.g., "optionally substituted with one, two or more R" means may be unsubstituted (unsubstituted) or may optionally be substituted with one, two or more R.
The term "halogen" denotes fluorine, chlorine, bromine and iodine.
The term "C 1-40 Alkyl is understood to meanA straight or branched chain saturated monovalent hydrocarbon group having 1 to 40 carbon atoms. For example, "C 1-10 Alkyl "denotes straight-chain and branched alkyl groups having 1,2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms," C 1-8 Alkyl "denotes straight and branched chain alkyl groups having 1,2, 3, 4, 5, 6, 7, or 8 carbon atoms," C 1-6 Alkyl "denotes straight-chain and branched alkyl groups having 1,2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof.
The term "C 2-40 Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more double bonds and having from 2 to 40 carbon atoms, preferably" C 2-10 Alkenyl ". "C 2-10 Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably" C 2-8 Alkenyl ". "C 2-10 Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms, for example having 2, 3, 4, 5 or 6 carbon atoms (i.e. C) 2-6 Alkenyl) having 2 or 3 carbon atoms (i.e., C) 2-3 Alkenyl). It is understood that in the case where the alkenyl group contains more than one double bond, the double bonds may be separated or conjugated to each other. The alkenyl group is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, (E) -but-2-enyl, (Z) -but-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hexa-vinyl5-alkenyl, (E) -hex-4-alkenyl, (Z) -hex-4-alkenyl, (E) -hex-3-alkenyl, (Z) -hex-3-alkenyl, (E) -hex-2-alkenyl, (Z) -hex-2-alkenyl, (E) -hex-1-alkenyl, (Z) -hex-1-alkenyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2-enyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E) -1-methylbut-2-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.
The term "C 2-40 Alkynyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more triple bonds and having from 2 to 40 carbon atoms, preferably" C 2-10 Alkynyl ". The term "C 2-10 Alkynyl "is understood as preferably meaning a straight-chain or branched monovalent hydrocarbon radical which contains one or more triple bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, for example 2, 3, 4, 5, 6, 7 or 8 carbon atoms (i.e.," C 2-8 Alkynyl ") having 2, 3, 4, 5, or 6 carbon atoms (i.e.," C 2-6 Alkynyl ") having 2 or 3 carbon atoms (" C) 2-3 Alkynyl "). The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl -alkynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbut-3-ynyl, 1,1-dimethylbut-2-ynyl or 3,3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.
The term "C 3-40 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) hydrocarbon or tricyclic hydrocarbon ring having 3 to 40 carbon atoms, preferably" C 3-10 Cycloalkyl group ", more preferably" C 3-8 Cycloalkyl groups ". The term "C 3-10 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic (e.g. bridged, spiro) hydrocarbon ring or tricycloalkane having 3,4, 5, 6, 7, 8, 9 or 10 carbon atoms. Said C is 3-10 Cycloalkyl can be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or bicyclic, such as bornyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo [2.1.1]Hexyl, bicyclo [2.2.1]Heptyl, bicyclo [2.2.1]Heptenyl, 6,6-dimethylbicyclo [3.1.1]Heptyl, 2,6,6-trimethylbicyclo [3.1.1]Heptyl, bicyclo [2.2.2]Octyl, 2,7-diazaspiro [3,5]Nonanyl, 2,6-diazaspiro [3,4 ]An octyl group, or a tricyclic hydrocarbon group such as an adamantyl group.
Unless otherwise defined, the term "3-20 membered heterocyclyl" means a saturated or unsaturated non-aromatic ring or ring system, e.g., which is a 4-, 5-, 6-or 7-membered monocyclic, 7-, 8-, 9-, 10-, 11-or 12-membered bicyclic (e.g., fused, bridged, spiro) or 10-, 11-, 12-, 13-, 14-or 15-membered tricyclic ring system, and contains at least one, e.g., 1, 2, 3, 4, 5 or more heteroatoms selected from 8978 zft 8978 and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitroxides, -S (O) -or-S (O) 2 -state of (c). Preferably, the heterocyclic group may be selected from "3-10 membered heterocyclic group". The term "3-10 membered heterocyclyl" means a saturated or unsaturated non-aromatic ring or ring system and contains at least one heteroatom selected from O, S and N. The heterocyclic group may be attached to the molecule through any of the carbon atoms or through a nitrogen atom (if present)The rest of the connection. The heterocyclic group may include fused or bridged rings as well as spiro rings. In particular, the heterocyclic group may include, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a 5,5 membered ring, such as hexahydrocyclopenta [ c ] ]Pyrrole-2 (1H) -cyclic rings, or 5,6 membered bicyclic rings, e.g., hexahydropyrrolo [1,2-a]A pyrazin-2 (1H) -yl ring. The heterocyclyl group may be partially unsaturated, i.e. it may contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H- [1,3,4]Thiadiazinyl, 4,5-dihydrooxazolyl, or 4H- [1,4]Thiazinyl, or it can be benzo-fused, such as but not limited to dihydroisoquinolinyl. When the 3-20 membered heterocyclic group is linked to another group to form the compound of the present invention, the carbon atom of the 3-20 membered heterocyclic group may be linked to another group, or the heterocyclic atom of the 3-20 membered heterocyclic ring may be linked to another group. For example, when the 3-20 membered heterocyclic group is selected from piperazinyl, it may be such that the nitrogen atom on the piperazinyl group is attached to another group. Or when the 3-20 membered heterocyclyl group is selected from piperidinyl, it may be that the nitrogen atom on the piperidinyl ring and the carbon atom in the para position are attached to other groups.
The term "C 6-20 Aryl "is understood to mean preferably a mono-, bi-or tricyclic hydrocarbon ring of monovalent or partially aromatic character having 6 to 20 carbon atoms, which may be a monoaromatic or polyaromatic ring fused together, preferably" C 6-14 Aryl ". The term "C 6-14 Aryl "is understood as preferably meaning a monocyclic, bicyclic or tricyclic hydrocarbon ring of monovalent or partial aromaticity having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" C) 6-14 Aryl group "), in particular a ring having 6 carbon atoms (" C 6 Aryl radicals ") For example phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C 9 Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C) 10 Aryl radicals), such as tetralinyl, dihydronaphthyl or naphthyl, or rings having 13 carbon atoms ("C 13 Aryl radicals), such as the fluorenyl radical, or a ring having 14 carbon atoms ("C) 14 Aryl), such as anthracenyl. When said C is 6-20 When the aryl group is substituted, it may be mono-or polysubstituted. And, the substitution site thereof is not limited, and may be, for example, ortho-, para-or meta-substitution.
The term "5-20 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic (e.g., fused, bridged, spiro) or tricyclic aromatic ring systems: having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, e.g., "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: which have 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and, in addition, in each case can be benzo-fused. "heteroaryl" also refers to a group in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclic rings, where the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-or 7-indolyl, 2-, 3-, 4-, 5-, 6-or 7-indazolyl, 2-, 4-, 5-, 6-, 7-or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7-or 8-phthalazinyl (phthalazinyl), 2-, 3-, 4-, 5-or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-or 8-cinnolinyl, 2-, 4-, 6-, 7-or 8-pteridinyl, 2-, 4-, 6-, 7-or 4-, 6-pteridinyl, 2-, 4-, 3-, 4-, 6-, 7-or 8-pteridinyl, 2-, 4-, 3-, 4-, 6-, 7-or 8-pteridinyl, 4-, 5-, 6-, 7-or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-or 9-carbolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-or 9-pyridyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-or 10-phenanthrolinyl, 1-, 2-, 1-, 6-, 8-, or 10-phenanthrolinyl 3-, 4-, 6-, 7-, 8-or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-or 10-phenazinyl, 2-, 3-, 4-, 5-, 6-or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-or 10-benzisoquinolinyl, 2-, 3-, 4-or thieno [ 8978 zft 8978-b ] furyl, 2-, 3-, 5-, (a) phenothiazinyl, a 1-, 2-, 3-, 7-, 8-, 9-, or 10-benzothioxanil group, 6-, 7-, 8-, 9-, 10-or 11-7H-pyrazino [2,3-c ] carbazolyl, 2-, 3-, 5-, 6-or 7-2H-furo [3,2-b ] -pyranyl, 2-, 3-, 4-, 5-, 7-or 8-5H-pyrido [2,3-d ] -o-oxazinyl, 1-, 3-or 5-1H-pyrazolo [4,3-d ] -oxazolyl, 2-, 4-or 54H-imidazo [4,5-d ] thiazolyl, 3-, 5-or 8-pyrazino [2,3-d ] pyridazinyl, 2-, 3-, 5-or 6-imidazo [2,1-b ] thiazolyl, 2-, 3-, 5-or 6-imidazo [4,5-d ] thiazolyl 1-, 3-, 6-, 7-, 8-or 9-furo [3,4-c ] cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10-or 11-4H-pyrido [2,3-c ] carbazolyl, 2-, 3-, 6-or 7-imidazo [1,2-b ] [1,2,4] triazinyl, 7-benzo [ b ] thienyl, 2-, 4-, 5-, 6-or 7-benzoxazolyl, 2-, 4-, 5-, 6-or 7-benzimidazolyl, 2-, 4-, 5-, 6-or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-or 9-benzoxazolyl (benzoxapinyl), 2-, 4-, 5-, 6-, 7-or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-or 11-1H-pyrrolo [ 8978 zft 8978-b ] [2] benzazepinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo [ b ] thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. . When the 5-20 membered heteroaryl group is linked to another group to form the compound of the present invention, the carbon atom on the 5-20 membered heteroaryl ring may be linked to another group, or the heteroatom on the 5-20 membered heteroaryl ring may be linked to another group. When the 5-20 membered heteroaryl group is substituted, it may be mono-or poly-substituted. Also, there is no limitation on the substitution site thereof, and for example, hydrogen attached to a carbon atom on a heteroaryl ring may be substituted, or hydrogen attached to a heteroatom on a heteroaryl ring may be substituted.
The term "spiro" refers to a ring system in which two rings share 1 ring atom.
The term "fused ring" refers to a ring system in which two rings share 2 ring atoms.
The term "bridged ring" refers to a ring system in which two rings share more than 3 ring-forming atoms.
Unless otherwise indicated, heterocyclyl, heteroaryl or heteroarylene include all possible isomeric forms thereof, e.g., positional isomers thereof. Thus, for some illustrative non-limiting examples, forms may be included that are substituted at 1, 2 or more of their 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) or bonded to other groups, including pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.
The term "oxo" refers to an oxy substitution (= O) formed after oxidation of a carbon atom, a nitrogen atom, or a sulfur atom in a substituent.
"hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
"THF" refers to tetrahydrofuran. "EtOAc" refers to ethyl acetate. "MeOH" refers to methanol. "DMF" refers to N, N-dimethylformamide. "DIPEA" refers to N, N-diisopropylethylamine. "TFA" refers to trifluoroacetic acid. "MeCN" refers to acetonitrile. "DMA" refers to N, N-dimethylacetamide. "Et 2 O "means diethyl ether. "DCE" refers to 1,2 dichloroethane. "NBS" refers to N-bromosuccinimide. "NIS" refers to N-iodosuccinimide. "Cbz-Cl" refers to benzyl chloroformate. Pd 2 (dba) 3 "refers to tris (dibenzylideneacetone) dipalladium. "Dppf" refers to 1,1' -bisdiphenylphosphinoferrocene. "HATU" refers to 2- (7-benzotriazole oxide) -N, N, N ', N' -tetramethyluronium hexafluorophosphate. "KHMDS refers to potassium hexamethyldisilazide. "LiHMDS" refers to lithium bistrimethylsilyl amide. "MeLi" refers to methyllithium. "n-BuLi" refers to n-butyllithium. "NaBH (OAc) 3 "refers to sodium triacetoxyborohydride.
Unless otherwise indicated, the definitions of terms herein apply equally to groups comprising the term, e.g. C 1-6 The definition of alkyl also applies to C 1-6 Alkyloxy, C 3-8 cycloalkyl-C 1-6 Alkyl-, and the like.
It will be appreciated by those skilled in the art that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; these compounds may also form inner salts if they contain both an acidic centre (e.g. carboxyl) and a basic centre (e.g. amino).
The compounds of the invention may be present in the form of solvates, such as hydrates, wherein the compounds of the invention comprise as structural element of the crystal lattice of the compound a polar solvent, such as in particular water, methanol or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric proportions.
Depending on their molecular structure, the compounds of the invention may be chiral and may therefore exist in various enantiomeric forms. These compounds may thus be present in racemic or optically active form. The compound of the invention covers isomers or mixtures and racemates thereof, wherein each chiral carbon is in R or S configuration. The compounds of the invention or intermediates thereof may be separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from mixtures by reaction with optically active resolving agents. Examples of suitable resolving agents are optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulphonic acids. The chromatographic enantiomeric resolution can also advantageously be carried out with the aid of optically active resolving agents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers, which are immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.
The corresponding stable isomers can be isolated according to known methods, for example by extraction, filtration or column chromatography.
The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses or primates, most preferably humans.
The term "therapeutically effective amount" means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought by a researcher, veterinarian, medical doctor or other clinician in a tissue, system, animal, individual, or human, which includes one or more of the following: (1) prevention of diseases: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not experienced or developed disease pathology or symptomatology. (2) inhibition of diseases: for example, inhibiting the disease, disorder or condition (i.e., arresting the further development of the pathology and/or condition) in an individual who is experiencing or presenting the pathology or condition of the disease, disorder or condition. (3) relieving the diseases: such as relieving the disease, disorder or condition (i.e., reversing the pathology and/or symptomatology) in an individual who is experiencing or developing the pathology or symptomatology of the disease, disorder or condition.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.
Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.
The structure of the compounds of the invention is determined by Nuclear Magnetic Resonance (NMR) or/and liquid mass chromatography (LC-MS). NMR chemical shifts (δ) are given in units of parts per million (ppm). NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d) 6 ) Deuterated methanol (CD) 3 OD) and deuterated chloroform (CDCl) 3 ) Internal standard is Tetramethylsilane (TMS).
LC-MS was measured using an Agilent 1200Infinity Series Mass spectrometer. HPLC was carried out using an Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18X 4.6mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18X 4.6mm column).
The thin layer chromatography silica gel plate adopts a tobacco yellow sea HSGF254 or Qingdao GF254 silica gel plate, the specification adopted by TLC is 0.15 mm-0.20 mm, and the specification adopted by the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm. The column chromatography uses 200-300 mesh silica gel of Futai Huanghai silica gel as a carrier.
All reactions of the present invention are carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, without specific indication, the solvent is a dried solvent, and the reaction temperature is given in degrees centigrade.
Example 1
Preparation of 3- [ 2-fluoro-4-methoxy-5- (quinolin-6-yl-methoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-001)
Figure BDA0003608368340000201
First step of
Preparation of 6- (bromomethyl) quinoline
Hydroxymethylquinoline 029a (500mg, 3.14mmol) and phosphine tribromide (1.28g, 47mmol) were dissolved in 1,4-dioxane (20 mL). The reaction solution was reacted at 25 ℃ for 0.5h. After completion of the reaction, the pH was adjusted to 7-8 with saturated sodium bicarbonate solution, followed by extraction with dichloromethane (2X 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 6- (bromomethyl) quinoline 029b (615 mg, yellow solid), yield: 79 percent.
MS m/z(ESI)：221.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.93(dd,J＝4.2,1.6Hz,1H),8.13(dd,J＝15.6,8.4Hz,2H),7.83(d,J＝2.0Hz,1H),7.76(dd,J＝8.8,2.0Hz,1H),7.44(dd,J＝8.4,4.4Hz,1H),4.68(s,2H).
Second step of
Preparation of 6- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline
4-fluoro-2-methoxy-5-nitrophenol 029c (300mg, 1.6 mmol) and 6- (bromomethyl) quinoline 029b (391mg, 1.76mmol) were dissolved in acetonitrile (20 mL), and potassium carbonate (422mg, 3.2mmol) was added thereto. The reaction mixture was reacted at 60 ℃ for 1 hour. Then extracted with dichloromethane (2X 60 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated to give a crude product which was purified over silica gel column (petroleum ether/ethyl acetate = 5/1) to give 6- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 029d (430 mg, yellow solid) in 75.31% yield.
MS m/z(ESI)：329.1[M+1] + 。
1 H NMR(400MHz,CD 3 OD)δ8.95(dd,J＝4.2,1.6Hz,1H),8.21(t,J＝9.2Hz,2H),7.92(s,1H),7.82(dd,J＝8.8,1.8Hz,1H),7.72(d,J＝7.2Hz,1H),7.47(dd,J＝8.2,4.4Hz,1H),6.77(d,J＝12.4Hz,1H),5.34(s,2H),3.99(s,3H).
The third step
Preparation of 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) aniline
6- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 029d (200mg, 6.1mmol) and iron powder (85.1636 mg,1.525 mmol) were dissolved in acetonitrile (20 mL) and a saturated ammonium chloride solution (5 mL) was added thereto. The reaction mixture was reacted at 25 ℃ for 16 hours. The reaction mixture was filtered to remove iron powder, and then extracted with dichloromethane (2X 60 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) aniline 029e (166 mg, red solid) in 82.10%.
MS m/z(ESI)：299.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.92(dd,J＝4.2,1.6Hz,1H),8.15(dd,J＝13.4,8.4Hz,2H),7.87(s,1H),7.77(dd,J＝8.8,1.6Hz,1H),7.42(dd,J＝8.4,4.4Hz,1H),6.68(d,J＝12.0Hz,1H),6.42(d,J＝8.8Hz,1H),5.25(s,2H),3.82(s,3H).
The fourth step
Preparation of 4- ({ [ 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) aniline 029e (20mg, 0.07mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (25.82mg, 0.077mmol) were dissolved in tetrahydrofuran (2 mL), triethylamine (21.21mg, 0.21mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 24 hours. This step was used directly in the next reaction without work-up.
MS m/z(ESI)：540.12[M+1] + 。
The fifth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (2 mL) were added to the reaction solution in the previous step, followed by addition of lithium hydroxide monohydrate (16.99mg, 0.405mmol). The reaction mixture was reacted at 25 ℃ for 1 hour. After completion of the reaction, the pH was adjusted to 6-7 with 1N hydrochloric acid, and the mixture was extracted with dichloromethane (2X 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The residue obtained was purified by preparative (acetonitrile/water/0.05% formic acid) to give 3- [ 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-001 (80 mg, light yellow solid) in 57% yield.
MS m/z(ESI)：494.1[M+1] + 。
HPLC:98.17％(214nm),95.65％(254nm).
1 H NMR(400MHz,d6-DMSO)δ14.54(s,1H),12.01(s,1H),8.92(dd,J＝4.2,1.6Hz,1H),8.38(d,J＝7.2Hz,1H),8.05(dd,J＝5.2,3.6Hz,2H),7.84(dd,J＝8.8,1.6Hz,1H),7.61-7.55(m,1H),7.34(d,J＝7.2Hz,1H),7.17(d,J＝11.6Hz,1H),5.21(s,2H),3.87(s,3H).
19 F NMR(376MHz,d6-DMSO)δ-128.11.
Example 2
Preparation of 3- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-002)
Figure BDA0003608368340000211
First step of
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxybenzene
4-fluoro-2-methoxyphenol 30a (2g, 14mmol) was added to acetone (20 mL), benzyl bromide (2.9g, 17mmol) and potassium carbonate (2.92g, 21.11mmol) were added, and the reaction was stirred at 65 ℃ for 16 hours. After completion of the reaction, water (50 mL) was added to the reaction liquid, extracted three times with ethyl acetate, the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1) to obtain 1- (benzyloxy) -4-fluoro-2-methoxybenzene 30b (3 g, yellow solid) in yield: 83 percent.
MS m/z(ESI)：233.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.46-7.29(m,5H),6.81(dd,J＝8.8,5.6Hz,1H),6.66(dd,J＝10.4,2.8Hz,1H),6.53(td,J＝8.5,2.8Hz,1H),5.11(s,2H),3.87(s,3H).
Second step of
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene
1- (benzyloxy) -4-fluoro-2-methoxybenzene 30b was dissolved in acetic anhydride (3 mL), cooled to 0 ℃, nitric acid (0.2mL, 65%) was added, and stirred at 0 ℃ for 4 hours, water (10 mL) was added to the reaction mixture after completion of the reaction, extracted three times with dichloromethane, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene 30c (550 mg, pale yellow solid) in yield: 83 percent.
MS m/z(ESI)：278.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.64(d,J＝7.2Hz,1H),7.47-7.37(m,5H),6.74(d,J＝12.4Hz,1H),5.15(s,2H),3.95(s,3H).
The third step
Preparation of 4-fluoro-2-methoxy-5-nitrophenol
1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene 30c (550mg, 1.98mmol) was dissolved in boron trichloride (3mL, 1M in DCM) and the reaction was stirred for 2 h at 25 ℃. After completion of the reaction, water (10 mL) was added to the reaction solution by LCMS, followed by dichloromethane (3 × 10 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain 4-fluoro-2-methoxy-5-nitrophenol 30d (200 mg, yellow solid) in yield: 51 percent.
MS m/z(ESI)：188.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.66(d,J＝7.2Hz,1H),6.74(d,J＝11.6Hz,1H),3.00(s,1H).
The fourth step
Preparation of isoquinolin-8-yl methanol
Isoquinolin-8-yl-carbaldehyde 30e (250mg, 1.59mmol) was dissolved in methanol (10 mL), sodium borohydride (30mg, 0.80mmol) was added, and the reaction was stirred at 25 ℃ for 4 hours. After completion of the reaction, water (10 mL) was added to the reaction solution, and extracted with dichloromethane (3 × 20 mL), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give isoquinolin-8-ylcarbinol 30f (210 mg, yellow oil) in yield: 75 percent.
MS m/z(ESI)：160.2[M+1] + 。
The fifth step
Preparation of 8- (chloromethyl) isoquinoline
Isoquinolin-8-ylcarbinol 30f (210mg, 1.38mmol) was dissolved in dichloromethane (20 mL), tsCl (395mg, 2.07mmol), DMAP (50mg, 0.41mmol) and triethylamine (419mg, 4.14mmol) were added, and the reaction solution was stirred at 25 ℃ for 16 hours. After completion of the reaction, water (20 mL) was added to the reaction solution, extracted with dichloromethane (3 × 20 mL), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give 30g (200 mg, pale yellow oil) of 8- (chloromethyl) isoquinoline, yield: 73 percent.
MS m/z(ESI)：178.2[M+1] + 。
The sixth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
30g (200mg, 1.13mmol) of 8- (chloromethyl) isoquinoline was dissolved in acetonitrile (20 mL), 4-fluoro-2-methoxy-5-nitrophenol 30d (232mg, 1.24mmol), potassium carbonate (312mg, 2.26mmol) and potassium iodide (19mg, 0.11mmol) were added, and the reaction solution was stirred at 65 ℃ for 2 hours. After completion of the reaction, water (20 mL) was added to the reaction solution, extracted with dichloromethane (3 × 20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/2) to obtain 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline for 30h (150 mg, pale yellow solid), yield: 36 percent.
MS m/z(ESI)：329.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.59(s,1H),8.62(d,J＝5.6Hz,1H),7.91-7.84(m,1H),7.81(d,J＝7.2Hz,1H),7.72(dd,J＝11.6,5.4Hz,3H),6.76(d,J＝12.4Hz,1H),5.63(s,2H),3.93(s,3H).
Seventh step
Preparation of 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyaniline
8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline (130mg, 0.37mmol) was dissolved in acetonitrile (10 mL) for 30h, iron powder (62mg, 1.11mmol) and saturated ammonium chloride solution (5 mL) were added, and the reaction mixture was stirred at 25 ℃ for 3 hours. After completion of the reaction by LCMS, the solid in the reaction solution was filtered off, extracted with dichloromethane (3 × 20 mL), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyaniline 30i (100 mg, pale yellow solid 81%), yield: 81 percent.
MS m/z(ESI)：299.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.66(s,1H),8.60(d,J＝5.0Hz,1H),7.86-7.79(m,1H),7.76-7.65(m,3H),6.69(d,J＝12.0Hz,1H),6.51(d,J＝8.8Hz,1H),5.55(s,2H),3.81(s,3H).
Eighth step
4- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyaniline 30i (100mg, 0.34mmol) was dissolved in tetrahydrofuran (20 mL), 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (228mg, 0.68mmol) and triethylamine (103mg, 1.02mmol) were added, and the reaction was stirred at 25 ℃ for 16 hours. After completion of the reaction monitored by LCMS, the reaction was concentrated to give the compound 4- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 30j (120 mg, brown solid), yield: and 59 percent of the total weight of the solution.
MS m/z(ESI)：540.1[M+1] + 。
The ninth step
Preparation of 3- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 30j (120mg, 0.222mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL) and methanol (5 mL), lithium hydroxide (24mg, 0.56mmol) was added, and the reaction was stirred at 25 ℃ for 3 hours. After completion of the reaction monitored by LCMS, the reaction was adjusted to pH 5-6 with 1N HCl, extracted with ethyl acetate (3X 20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated, and the residue was prepared (acetonitrile/water/0.1% formic acid) to give 3- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-002 (30 mg, pale yellow solid), yield: 32 percent.
MS m/z(ESI)：494.2[M+1] + 。
HPLC:99.57％(214nm),97.09％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.52(s,1H),12.04(s,1H),9.70(s,1H),8.65(d,J＝6.0Hz,1H),8.16-8.11(m,2H),7.96-7.90(m,2H),7.46-7.40(m,2H),7.19(d,J＝11.6Hz,1H),5.62(s,2H),3.84(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-74.17,-127.58--127.79.
Examples 3 and 4
Preparation of 3- (5- { [ (4S) -5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-003) and 3- (5- { [ (4R) -5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-004)
Figure BDA0003608368340000241
First step of
3- (2-bromo-4,5-difluorophenoxy) propionic acid
To a solution of 2-bromo-4,5-difluorophenol 031a (5 g,23.9 mmol) and 3-bromopropionic acid (3.6 g,23.9 mmol) was added aqueous sodium hydroxide (2.1g, 52.5 mmol) solution (6 mL). The reaction mixture was stirred at 100 ℃ for 6 hours. Water (40 mL) was added. The mixture solution was acidified with hydrochloric acid (2M) to pH =1-2, and then extracted with ethyl acetate (3 × 50 mL). The combined organic phases are washed with Na 2 SO 4 Dried and then concentrated. The residue was purified by flash column chromatography to give 3- (2-bromo-4,5-difluorophenoxy) propionic acid 031b (3.6 g, yellow solid). Yield: 48 percent.
MS m/z(ESI)：263.0(M-OH)。
1 H NMR(400MHz,CDCl 3 )δ7.38(dd,J＝9.2,8.4Hz,1H),6.80(dd,J＝11.6,7.2Hz,1H),4.25(t,J＝6.0Hz,2H),2.92(t,J＝6.0Hz,2H).
Second step of
Preparation of 8-bromo-5,6-difluoro-2,3-dihydro-1-benzopyran-4-one
To a solution of 3- (2-bromo-4,5-difluorophenoxy) propionic acid 031b (3.6g, 13.5 mmol) in dichloromethane (100 mL) was added a catalytic amount of DMF. Oxalyl chloride (3.4 g, 27mmol) was slowly added dropwise to the reaction at 0 ℃. After the addition was complete, the reaction mixture was stirred at 25 ℃ for 1h. Aluminum trichloride (2g, 14.8mmol) was added at 0 ℃. The reaction mixture was stirred at 25 ℃ for 16h. Water (40 mL) was added and stirred for 20min. The mixture was extracted with dichloromethane (2X 50 mL). All organic phases were combined, the organic phase was washed with 0.5M aqueous NaOH (2X 50 mL), brine (2X 40 mL), dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 6/1) to obtain 8-bromo-5,6-difluoro-2,3-dihydro-1-benzopyran-4-one 031c (3.1 g, yellow solid) with a yield of 78%.
MS m/z(ESI)：263.0(M+H)。
1 H NMR(400MHz,CDCl 3 )δ7.64-7.59(dd,J＝9.2,8.0Hz,1H),4.65-4.62(m,2H),2.88-2.84(m,2H).
The third step
Preparation of 8-bromo-5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-ol
8-bromo-5,6-difluoro-2,3-dihydro-1-benzopyran-4-one 031c (0.5g, 1.9mmol) was dissolved in MeOH (5 mL) to which was slowly added sodium borohydride (108mg, 2.8mmol). The reaction solution was stirred at 25 ℃ for 15 minutes. The reaction solution was acidified with 2M hydrochloric acid to pH =6-7 and extracted with ethyl acetate (3 × 10 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 8-bromo-5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-ol 031d (0.5 g, yellow solid). Yield: 89 percent.
MS m/z(ESI)：247.0(M-OH)。
1 H NMR(400MHz,CDCl 3 )δ7.37(t,J＝9.0Hz,1H),5.09(s,1H),4.45(d,J＝10.8Hz,1H),4.30(t,J＝11.6Hz,1H),3.74(s,1H),2.13-1.99(m,2H).
The fourth step
Preparation of 8-bromo-5,6-difluoro-4- (4-fluoro-5-methoxy-2-nitrophenoxy) -3,4-dihydro-2H-1-benzopyran
To a solution of 8-bromo-5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-ol 031d (450mg, 1.7 mmol) in THF (5 mL) was added 4-fluoro-2-methoxy-5-nitrophenol (318mg, 1.7 mmol) and triphenylphosphine (580mg, 2.21mmol). DIAD (447mg, 2.21mmol) was added slowly at 0 ℃. The reaction was then stirred at 45 ℃ for 1h. The reaction mixture was concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate =10/1 to 5/1) to give 8-bromo-5,6-difluoro-4- (4-fluoro-5-methoxy-2-nitrophenoxy) -3,4-dihydro-2H-1-benzopyran 031e (800 mg, yellow solid). Yield: 86 percent.
MS m/z(ESI)：456.0(M+Na)。
1 H NMR(400MHz,CDCl 3 )δ7.79(d,J＝7.6Hz,1H),7.46-7.39(m,1H),6.77(d,J＝12.4Hz,1H),5.55(s,1H),4.56-4.42(m,2H),3.92(s,3H),2.34-2.30(m,1H),2.13-2.06(m,1H).
The fifth step
Preparation of 2- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -5-fluoro-4-methoxyaniline
To a solution of 8-bromo-5,6-difluoro-4- (4-fluoro-5-methoxy-2-nitrophenoxy) -3,4-dihydro-2H-1-benzopyran 031e (690mg, 1.59mmol) in methanol/ethyl acetate =2/1 (40 mL) was added 10% pd/C (100 mg). The reaction solution was replaced with hydrogen three times and stirred at 25 ℃ for 5 hours. The reaction mixture was filtered and concentrated. The obtained residue was separated and purified by a silica gel column (dichloromethane/methanol =100/1 to 10/1) to give 2- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -5-fluoro-4-methoxyaniline 031f (380 mg, brown solid). Yield: 66 percent.
MS m/z(ESI)：326.1(M+H)。
1 H NMR(400MHz,CDCl 3 )δ7.29(s,1H),7.04(dd,J＝18.4,9.2Hz,1H),6.75(d,J＝8.4Hz,1H),6.60(d,J＝6.8Hz,1H),5.43(s,1H),4.42-4.23(m,2H),3.80(s,3H),2.25(d,J＝12.0Hz,1H),1.96(s,1H).
The sixth step
Preparation of dimethyl 4- [ ({ 5- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
To a solution of 5- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -2-fluoro-4-methoxyaniline 031f (100mg, 0.31mmol) in THF (2 mL) was added 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (104mg, 0.31mmol) and triethylamine (94mg, 0.93mmol). The reaction mixture was stirred at 25 ℃ for 6h. The reaction mixture was concentrated. The resulting residue was separated and purified by silica gel column (petroleum ether/ethyl acetate =6/1 to 3/1) to give 4- [ ({ 5- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 031g (180 mg, yellow solid). Yield: 87 percent.
MS m/z(ESI)：567.1(M+H)。
1 H NMR(400MHz,CDCl 3 )δ8.79(d,J＝14.4Hz,1H),7.93(d,J＝5.2Hz,1H),7.67(d,J＝8.0Hz,1H),7.05(dd,J＝18.8,9.2Hz,1H),6.74(d,J＝12Hz,1H),6.63-6.61(m,1H),6.50(s,1H),5.50(s,1H),4.52-4.42(m,1H),4.35-4.25(m,1H),3.95-3.78(m,9H),2.29(dd,J＝14.8,2.0Hz,1H),2.01-1.90(m,1H).
Seventh step
Preparation of 3- (5- { [ (4S) -5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of 4- [ ({ 5- [ (5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl) oxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 031g (24mg, 0.04mmol) in tetrahydrofuran/methanol/water =1 (6 mL) was added lithium hydroxide (4 mg, 0.16mmol). The reaction mixture was stirred at 25 ℃ for 1h. The reaction solution was acidified with hydrochloric acid (2M) to pH =6-7 and extracted with dichloromethane (3 × 15 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residual oil was first subjected to chiral preparative HPLC (column: chiralpak-OD, mobile phase carbon dioxide-methanol (0.1% dea)), then lyophilized and purified by conventional preparative HPLC (column: -Xbridge-C18 × 19mm.5um, mobile phase acetonitrile-water (0.1% formic acid)) to give 3- (5- { [ (4S) -5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-003 (4 mg, near white solid) and 3- (5- { [ (4R) -5,6-difluoro-3,4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -25 zxft-2H-1-benzopyran-4-yl ] oxy } -5732, yield near white solid [ 3432H-5732): 19 percent.
Cpd-003:
MS m/z(ESI)：521.1(M+H)。
1 H NMR(400MHz,d 6 -DMSO)δ14.54(s,1H),12.00(d,J＝4.0Hz,1H),7.47-7.27(m,3H),7.18(d,J＝11.2Hz,1H),6.79-6.70(m,1H),5.45(s,1H),4.34(d,J＝10.6Hz,1H),4.16(t,J＝11.6Hz,1H),3.83(d,J＝2.8Hz,3H),2.16-2.12(m,1H),2.01-1.93(m,1H).
Cpd-004:
MS m/z(ESI)：521.0(M+H)。
1 H NMR(400MHz,CD 3 OD)δ7.31(s,1H),7.24-7.07(m,2H),7.05-7.01(dd,J＝11.2,3.2Hz,1H),6.67-6.58(m,1H),5.51(s,1H),4.36-4.25(m,2H),3.88(d,J＝9.2Hz,3H),2.28-2.25(m,1H),1.99-1.88(m,1H).
Example 5
Preparation of 3- {4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-005)
Figure BDA0003608368340000261
First step of
Preparation of 2- (benzyloxy) -4-fluoro-1-methoxybenzene
5-fluoro-2-methoxyphenol 035a (1 g,7 mmol) was dissolved in N, N-dimethylformamide (10 mL), and benzyl bromide (1.44g, 8.4mmol) and potassium carbonate (2.91g, 21mmol) were added. The reaction mixture was stirred at 80 ℃ for 6 hours. The reaction was quenched with water (100 mL) and extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 4/1) to give the title product 2- (benzyloxy) -4-fluoro-1-methoxybenzene 035b (1.5 g, white solid) with a yield of 87%.
MS m/z(ESI)：255.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ7.43(d,J＝7.2Hz,2H),7.40-7.34(m,2H),7.34-7.29(m,1H),6.81(dd,J＝8.8,5.2Hz,1H),6.65(dd,J＝10.0,2.8Hz,1H),6.63-6.57(m,1H),5.13(s,2H),3.86(s,3H).
Second step of
Preparation of 1- (benzyloxy) -5-fluoro-2-methoxy-4-nitrobenzene
2- (benzyloxy) -4-fluoro-1-methoxybenzene 035b (100mg, 0.43mmol) was dissolved in acetic anhydride (3 mL), and 65% nitric acid (109mg, 1.72mmol) was added dropwise under an ice-water bath. The reaction solution was stirred at 25 ℃ for 12 hours. The reaction solution was adjusted to pH 7 with sodium hydroxide solution, followed by extraction with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 4/1) to give the title product 1- (benzyloxy) -5-fluoro-2-methoxy-4-nitrobenzene 035c (110 mg, white solid) with a yield of 83%.
MS m/z(ESI)：278.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ7.59(d,J＝7.2Hz,1H),7.46-7.35(m,5H),6.75(d,J＝12.4Hz,1H),5.21(s,2H),3.94(s,3H).
The third step
Preparation of 5-fluoro-2-methoxy-4-nitrophenol
1- (benzyloxy) -5-fluoro-2-methoxy-4-nitrobenzene 035c (1g, 3.6mmol) was dissolved in 1M boron trichloride in dichloromethane (10.8mL, 10.8mmol). The reaction mixture was stirred at 25 ℃ for 12 hours. The reaction solution was quenched with methanol (10 mL), and the concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 3/2) to give the title product 5-fluoro-2-methoxy-4-nitrophenol 035d (0.6 g, yellow solid) with a yield of 80%.
MS m/z(ESI)：188.1[M+1] + .
1 H NMR(400MHz,DMSO)δ11.29(s,1H),7.62(d,J＝7.6Hz,1H),6.86(d,J＝12.8Hz,1H),3.86(s,3H).
The fourth step
Preparation of 2,3-difluoro-6-methoxybenzaldehyde
A2M lithium diisopropylamide solution (3.82mL, 7.64mmol) was dissolved in tetrahydrofuran (10 mL), a solution of 1,2-difluoro-4-methoxybenzene 035e (1000mg, 6.94mmol) in tetrahydrofuran (3 mL) was added dropwise at-78 deg.C, and after stirring at-78 deg.C for 1 hour, N-dimethylformamide (558mg, 7.64mmol) was added dropwise and further stirred for 1 hour. The reaction solution was quenched with acetic acid (1.5 mL) and water (50 mL), and then extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (dichloromethane/methanol = 30/1) to give the title product 2,3-difluoro-6-methoxybenzaldehyde 035f (1050 mg, yellow solid) in 79% yield.
MS m/z(ESI)：173.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ10.44-10.37(m,1H),7.34(dd,J＝18.0,9.2Hz,1H),6.72-6.68(m,1H),3.92(s,3H).
The fifth step
Preparation of 2-vinyl-3,4-difluoro-1-methoxybenzene
Tris (triphenylphosphine) methyl bromide (1.71g, 4.80mmol) was dissolved in tetrahydrofuran (15 mL) and cooled to-15 deg.C, and a solution of 1M potassium tert-butoxide (6.0 mL,6.00mmol, 3eq) in tetrahydrofuran was added dropwise. The reaction solution was stirred at 25 ℃ for 1 hour. After cooling to-15 ℃ a solution of 2,3-difluoro-6-methoxybenzaldehyde 035f (689mg, 4.00mmol) in tetrahydrofuran (3 mL) was added and the reaction was stirred at 25 ℃ for an additional 1 hour. The reaction was quenched with saturated ammonium chloride solution (100 mL) and extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 50/1) to give the title product 2-vinyl-3,4-difluoro-1-methoxybenzene 035g (4.25 g, light yellow oil) in 75% yield.
MS m/z(ESI)：171.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ7.02-6.95(m,1H),6.80(dd,J＝18.0,12.0Hz,1H),6.55(ddd,J＝9.2,3.6,2.0Hz,1H),6.05(dt,J＝18.0,1.6Hz,1H),5.56(dt,J＝12.0,1.6Hz,1H),3.83(s,3H).
The sixth step
Preparation of 2- (2,3-difluoro-6-methoxyphenyl) ethanol
2-vinyl-3,4-difluoro-1-methoxybenzene 035g (1000mg, 5.88mmol) was dissolved in tetrahydrofuran (5 mL), a solution of 0.5M 9-BBN (35mL, 17.64mmol) in tetrahydrofuran was added dropwise, the reaction mixture was stirred at 25 ℃ for 16 hours, and then 3N NaOH (9.8 mL) and H were added 2 O 2 (3332mg, 29.4 mmol). The reaction mixture was stirred at 25 ℃ for a further 1 hour. The reaction solution was quenched with water (100 mL) and the pH adjusted To 6, then extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The residue after concentration was purified by silica gel column (petroleum ether/ethyl acetate = 1/2) to give the title product 2- (2,3-difluoro-6-methoxyphenyl) ethanol 035h (800 mg, colorless oil) in 65% yield.
1 H NMR(400MHz,DMSO)δ7.23(dd,J＝19.6,9.2Hz,1H),6.79-6.76(m,1H),4.75(t,J＝5.2Hz,1H),3.79(s,3H),3.49(dd,J＝12.4,7.2Hz,2H),2.78-2.75(m,2H).
Seventh step
8978 preparation of zxft 8978-difluoro-3- [2- (5-fluoro-2-methoxy-4-nitrophenoxy) ethyl ] -4-methoxybenzene
2- (2,3-difluoro-6-methoxyphenyl) ethanol 035h (250mg, 1.34mmol) was dissolved in tetrahydrofuran (5 mL), and 5-fluoro-2-methoxy-4-nitrophenol 035d (303mg, 1.61mmol), triphenylphosphine (703mg, 2.68mmol) and diisopropyl azodicarboxylate (542mg, 2.68mmol) were added in that order. The reaction solution was stirred in a microwave reactor at 60 ℃ for 1 hour. The reaction was concentrated, and the resulting residue was purified by reverse phase column (acetonitrile/water = 3/2) to give the title product 1,2-difluoro-3- [2- (5-fluoro-2-methoxy-4-nitrophenoxy) ethyl ] -4-methoxybenzene 035i (220 mg, light yellow solid) in 41% yield.
MS m/z(ESI)：358.1[M+1] + .
1 H NMR(400MHz,DMSO)δ7.61(d,J＝7.2Hz,1H),7.32-7.27(m,2H),6.86-6.81(m,1H),4.27(t,J＝7.2Hz,2H),3.82(s,3H),3.80(s,3H),3.09(t,J＝6.4Hz,2H).
Eighth step
Preparation of 4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyaniline
1,2-difluoro-3- [2- (5-fluoro-2-methoxy-4-nitrophenoxy) ethyl ] -4-methoxybenzene 035i (200mg, 0.56mmol) was dissolved in methanol (10 mL) and wet palladium on charcoal (60mg, 0.56mmol) was added. The reaction solution was stirred at 25 ℃ for 6 hours under a hydrogen atmosphere. Filtration and concentration of the filtrate gave a residue which was purified by silica gel column (petroleum ether/ethyl acetate = 1/2) to give the title product 4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyaniline 035j (160 mg, light brown oil) in 78% yield.
MS m/z(ESI)：328.2[M+1] + .
1 H NMR(400MHz,DMSO)δ7.32-7.24(m,1H),6.84-6.78(m,1H),6.74-6.69(m,1H),6.44-6.38(m,1H),4.67(s,2H),3.94(t,J＝7.2Hz,2H),3.81(s,3H),3.62(s,3H),2.98(dd,J＝7.2,6.0Hz,2H).
The ninth step
Preparation of 4- [ ({ 4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyaniline 035j (130mg, 0.40mmol) was dissolved in tetrahydrofuran (5 mL), 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (135mg, 0.4 mmol) and triethylamine (122mg, 1.2mmol) were added sequentially, and the reaction was stirred at 25 ℃ for 16 hours, the reaction was concentrated to give the title product 4- [ ({ 4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 035k (180 mg, yellow oil) with a yield of 71%.
MS m/z(ESI)：569.1[M+1] + .
The tenth step
Preparation of 3- {4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ 4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 035k (200mg, 0.35mmol) was dissolved in tetrahydrofuran (4 mL), lithium hydroxide monohydrate (74mg, 1.75mmol) was added, the reaction was stirred at 25 ℃ for 1 hour then water (1 mL) was added, the reaction was stirred at 25 ℃ for 2 hours, the pH was adjusted to 6 with 1N hydrochloric acid solution, then ethyl acetate (3X 50 mL) was extracted, the organic phases were combined and washed with saturated brine, anhydrous sodium sulfate was dried, the concentrated residue was purified by preparative HPLC (acetonitrile/water (0.1% trifluoroacetic acid) to give the title product 3- {4- [2- (2,3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } -3245 zxft-dioxo-6-thiophene-1H-1-pyridine-1-yellow carboxylic acid yield (32 mg, 31 g: 32 d) and yellow yield of p-pyridine-3748 g.
MS m/z(ESI)：523.0[M+1] + .
HPLC:100％(214nm),99.06％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.55(s,1H),11.99(s,1H),7.39(s,1H),7.31(dd,J＝19.6,9.2Hz,1H),7.16-7.10(m,2H),6.87-6.82(m,1H),4.18(t,J＝7.2Hz,2H),3.82(s,3H),3.68(s,3H),3.10(t,J＝6.8Hz,2H).
Example 6
Preparation of 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- {4- [ ((methoxycarbamoyl) amino ] phenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid (Cpd-006)
Figure BDA0003608368340000291
First step of
Preparation of 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline
1,2-difluoro-3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -4-methoxybenzene 047a (2.4g, 7.3mmol) was dissolved in acetonitrile (20 mL). Iron powder (1.22g, 21.9mmol) and saturated ammonium chloride solution were then added. The reaction mixture was reacted at 25 ℃ for 8 hours. Then extracted with dichloromethane (2X 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline 047b (2.0 g, brown solid) in 78.08% yield.
MS m/z(ESI)：314.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.08(dd,J＝18.4,9.2Hz,1H),6.69-6.45(m,3H),5.06(s,2H),3.80(s,3H),3.73(s,3H).
Second step of
Preparation of 2-amino-5-bromothiophene-3,4-dicarboxylic acid diethyl ester
2-aminothiophene-3,4-dicarboxylic acid diethyl ester 047c (330mg, 1.36mmol) was dissolved in dichloromethane (5 mL) and N-bromosuccinimide (265.57mg, 1.49mmol) was added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then extracted with dichloromethane (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-amino-5-bromothiophene-3,4-dicarboxylic acid diethyl ester 047d (398 mg, red solid) in 91.96% yield.
MS m/z(ESI)：322.0[M+1] + 。
1 H NMR(400MHz,CD 3 OD)δ4.33(q,J＝7.2Hz,2H),4.22(q,J＝7.2Hz,2H),1.36(t,J＝7.2Hz,3H),1.27(t,J＝7.2Hz,3H).
The third step
Preparation of 2-bromo-5- [ (phenoxycarbonyl) amino ] -3,4-dicarboxylic acid diethyl ester
2-amino-5-bromothiophene-3,4-dicarboxylic acid diethyl ester 047d (400mg, 1.24mmol) and phenyl chloroformate (582.4 mg, 3.72mmol) were dissolved in toluene (5 mL). The reaction mixture was reacted at 110 ℃ for 3 hours. After completion of the reaction, it was cooled to room temperature and concentrated to give a crude product, which was passed through a silica gel column (petroleum ether/ethyl acetate = 8/2) to give 2-bromo-5- [ (phenoxycarbonyl) amino ] -3,4-dicarboxylic acid diethyl ester 047e (306 mg, white solid) in 50.22% yield.
MS m/z(ESI)：422.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.40(t,J＝8.0Hz,2H),7.27(d,J＝7.6Hz,1H),7.19(d,J＝7.6Hz,2H),4.34(dq,J＝21.2,7.2Hz,4H),1.35(dt,J＝23.2,7.2Hz,6H).
The fourth step
Preparation of 2-bromo-5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-3,4-dicarboxylic acid diethyl ester
Diethyl 2-bromo-5- [ (phenoxycarbonyl) amino ] thiophene-3,4-dicarboxylate 047e (388mg, 0.878mmol) and 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline (250mg, 0.798mmol) were dissolved in tetrahydrofuran (10 mL), triethylamine (282.09mg, 2.79mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 24 hours. Concentration gave a crude product which was passed through a silica gel column (petroleum ether/ethyl acetate = 7/3) to give 2-bromo-5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-3,4-dicarboxylic acid diethyl ester 047f (380 mg, yellow solid) in 64.79% yield.
MS m/z(ESI)：663.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ10.67(s,1H),7.62(s,1H),7.10(dd,J＝12.0,7.2Hz,1H),6.71(d,J＝12.0Hz,1H),6.58(ddd,J＝9.2,3.6,2.0Hz,1H),5.18(d,J＝1.6Hz,2H),4.37(q,J＝7.2Hz,2H),4.24(q,J＝7.2Hz,2H),3.82(d,J＝5.2Hz,6H),1.39(t,J＝7.2Hz,3H),1.27(dd,J＝7.2,2.4Hz,3H).
The fifth step
Preparation of ethyl 6-bromo-3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
Reacting 2-bromo-5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy]-2-fluoro-4-methoxyphenyl } carbamoyl) amino]Thiophene-3,4-dicarboxylic acid diethyl ester 047f (180mg, 0.272mmol) and sodium methoxide (44.08mg, 0.82mmol) were dissolved in methanol (2 mL). The reaction mixture was reacted at 40 ℃ for 16 hours. After the reaction was completed, the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, and the mixture was quenched and extracted with dichloromethane (2X 5 mL). The organic phases are combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain a crude product, and the crude product is purified by silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain 6-bromo-3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [2,3-d]Pyrimidine-5-carboxylic acid ethyl ester 047g (110 mg, white solid), yield: 59.13%. MS m/z (ESI): 615.0[ M ] +1] + 。
The sixth step
Preparation of ethyl 6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
6-bromo-3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid ethyl ester 047g (110mg, 0.178mmol), (6-aminopyridin-3-yl) borandiol (49.32mg, 0.357mmol), potassium carbonate (74.14mg, 0.54mmol), tris (dibenzylideneacetone) dipalladium (49.12mg, 0.054mmol) and 2-bicyclic hexylphosphine-2 ',6' -diisopropoxybiphenyl (25.03mg, 0.0536mmol) were dissolved in toluene: ethanol: water =7:2:1 (1 mL). The reaction solution was subjected to microwave treatment at 80 ℃ for 30 minutes. After completion of the reaction, it was extracted with dichloromethane (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the crude product, which was purified over silica gel column (petroleum ether/ethyl acetate = 2/8) to give ethyl 6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 047H (47 mg, white solid) with 33.50% yield.
MS m/z(ESI)：628.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.23(d,J＝8.4Hz,2H),7.07(d,J＝9.2Hz,1H),6.98(d,J＝7.2Hz,1H),6.72(d,J＝10.8Hz,1H),6.65(d,J＝8.4Hz,2H),6.55(d,J＝9.2Hz,1H),5.11(q,J＝10.4Hz,2H),4.32(dd,J＝14.4,7.2Hz,2H),3.76(d,J＝3.2Hz,6H),1.26(t,J＝7.2Hz,3H).
Seventh step
Preparation of 6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid
Ethyl 6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 047H (44mg, 0.0701mmol) and sodium hydroxide (8.82mg, 0.21mmol) were dissolved in methanol: water =3:1 (2 mL). The reaction mixture was reacted at 40 ℃ for 16 hours. After completion of the reaction, the mixture was extracted with methylene chloride (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid 047i (30.0 mg, light yellow solid) in 64.19% yield.
MS m/z(ESI)：600.7[M+1] + 。
Eighth step
Preparation of 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- {4- [ ((methoxycarbamoyl) amino ] phenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid
6- (4-aminophenyl) -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid 047i (30mg, 0.05mmol) and 4-nitrophenyl-N-methoxycarbamate were dissolved in tetrahydrofuran (2 mL) and triethylamine (46mg, 0.45mmol) was added. The reaction mixture was reacted at 25 ℃ for 90 minutes. After completion of the reaction, the mixture was extracted with methylene chloride (2X 35 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give a crude product which was prepared (acetonitrile/water/0.01% formic acid) to give 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- {4- [ ((methoxycarbamoyl) amino ] phenyl } -2,4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid Cpd-006 (5.3 mg, white solid) in 15% yield.
MS m/z(ESI)：673.1[M+1] + 。
HPLC:95.91％(214nm),90.84％(254nm).
1 H NMR(400MHz,d6-DMSO)δ9.55(s,1H),9.01(s,1H),8.21(s,1H),7.64(d,J＝8.8Hz,2H),7.39(d,J＝8.4Hz,2H),7.12(d,J＝7.2Hz,1H),7.00(d,J＝11.6Hz,1H),6.83(d,J＝9.6Hz,1H),4.94(s,2H),3.76(d,J＝5.6Hz,6H),3.60(s,3H).
19 F NMR(376MHz,d6-DMSO)δ-128.13，-140.25，-148.13--148.37。
Example 7
Preparation of methyl 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate (Cpd-007)
Figure BDA0003608368340000311
First step of
Preparation of diethyl 2-amino-5- { [ (tert-butoxy) carbonyl ] amino } thiophene-3,4-dicarboxylate
2,5-diaminothiophene-3,4-dicarboxylic acid diethyl ester 48a (4.00g, 15.49mmol) was dissolved in dichloromethane (50 mL), followed by the addition of Boc anhydride (3.38g, 15.49mmol), triethylamine (4.70g, 46.47mmol), DMAP (189mg, 1.55mmol). The reaction solution was stirred at room temperature for 2 hours. LCMS detection the solvent was evaporated after completion of the reaction, and the crude product was purified by silica gel column (petroleum ether/ethyl acetate = 30%) and concentrated under reduced pressure to give the desired product diethyl 2-amino-5- { [ (tert-butoxy) carbonyl ] amino } thiophene-3,4-dicarboxylate 48b (2.00 g, white solid) in yield: 25 percent.
MS m/z(ESI)：359.1[M+1] + 。
Second step of
Preparation of diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ (phenoxycarbonyl) amino ] thiophene-3,4-dicarboxylate
Diethyl 2-amino-5- { [ (tert-butoxy) carbonyl ] amino } thiophene-3,4-dicarboxylate 48b (800mg, 2.23mmol) was dissolved in toluene (20 mL), followed by addition of phenyl chloroformate (1.04g, 6.69mmol), and stirring at 110 ℃ for 3h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove toluene. The crude product was purified by silica gel column (petroleum ether/ethyl acetate = 20%) and concentrated to give the desired product 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ (phenoxycarbonyl) amino ] thiophene-3,4-dicarboxylic acid diethyl ester 48c (800 mg, yellow solid), yield: 45 percent.
MS m/z(ESI)：479.2[M+1] + 。
The third step
Preparation of 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-3,4-dicarboxylic acid diethyl ester
Diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ (phenoxycarbonyl) amino ] thiophene-3,4-dicarboxylate 48c (600mg, 1.25mmol) and 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline 47b (470mg, 1.5 mmol) were dissolved in THF (20 mL), and triethylamine (380mg, 3.75mmol) was then added to the reaction solution. The reaction was stirred at room temperature for 16h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by silica gel column (petroleum ether/ethyl acetate = 30%) and concentrated to give diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-3,4-dicarboxylate 48d (300 mg, white solid) in 33% yield.
MS m/z(ESI)：698.2(M+23)。
The fourth step
Preparation of methyl 6- { [ (tert-butoxy) carbonyl ] amino } -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
2- { [ (tert-butoxy) carbonyl ] amino } -5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-3,4-dicarboxylic acid diethyl ester 48d (200mg, 0.29mmol) was dissolved in methanol (20 mL), followed by addition of sodium methoxide (47mg, 0.87mmol). The reaction solution was stirred at 40 ℃ for 48 hours, after completion of the reaction was cooled to room temperature, the reaction solution was adjusted to pH 6-7 with 1N hydrochloric acid, and then extracted with ethyl acetate (3X 30 mL). After the organic phases were combined, saturated brine was washed with water, dried over anhydrous sodium sulfate, the crude product after concentration, and after purification by silica gel column (petroleum ether/ethyl acetate = 40%), the crude product was concentrated to give 6- { [ (tert-butoxy) carbonyl ] amino } -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-1-pyrimidine [ 3732 ] methyl formate as a white solid in a yield of 70 mg: 3736 mg.
MS m/z(ESI)：638.1[M+1] + 。
The fifth step
Preparation of methyl 6-amino-3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
Methyl 6- { [ (tert-butoxy) carbonyl ] amino } -3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 48e (30mg, 0.08mmol) was dissolved in DCM (5 mL) and trifluoroacetic acid (1 mL) was added. The reaction was stirred at room temperature for 2h. After completion of the reaction the solvent was concentrated by evaporation to afford preparation 48f (23 mg, white solid) of methyl 6-amino-3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate, yield: 81 percent.
MS m/z(ESI)：538.1[M+1] + 。
The sixth step
Preparation of methyl 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylate
Preparation of methyl 6-amino-3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 48f (23mg, 0.04mmol) was dissolved in anhydrous tetrahydrofuran (9 mL) followed by addition of phenyl chloroformate (2mg, 0.13mmol). The reaction was stirred at 90 ℃ for 2h. After completion of the reaction, concentration under reduced pressure gave 48g (40 mg, brown oil) of methyl 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylate, yield: 71 percent.
MS m/z(ESI)：562.1[M+1] + 。
Seventh step
Preparation of methyl 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
48g (20mg, 0.03mmol) of methyl 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylate was dissolved in THF (10 mL), followed by addition of triethylamine (63mg, 0.62mmol) and methylhydroxylamine hydrochloride (26mg, 0.31mmol). The reaction was stirred at room temperature for 16h. After completion of the reaction the solvent was evaporated to give the crude product which was purified by preparative HPLC (0.1% formic acid/acetonitrile/water) to give 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester Cpd-007 (2 mg, white solid) yield: 10 percent.
MS m/z(ESI)：611.1[M+1] + 。
HPLC:98.94％(214nm),99.08％(254nm).
1 H NMR(400MHz,CD 3 OD)δ7.50-7.40(m,1H),7.23(dd,J＝19.6,9.2Hz,2H),7.03(d,J＝7.2Hz,1H),6.94(d,J＝11.2Hz,1H),6.80-6.76(m,1H),5.11(d,J＝1.4Hz,2H),3.87(s,3H),3.85(s,3H),3.81(d,J＝1.2Hz,6H).
19 F NMR(376MHz,CD 3 OD)δ-129.23,-142.22,-150.25--150.82.
Example 8
Preparation of 3- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-008)
Figure BDA0003608368340000331
First step of
Preparation of methyl 5-amino-4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-methoxy-5-nitrobenzoate 064a (1.00g, 4.36mmol) was dissolved in acetonitrile (30 mL) and saturated ammonium chloride solution (10 mL), followed by addition of iron powder (2.44g, 43.63mmol). The reaction solution was stirred at room temperature for 4 hours. After LCMS detection reaction was complete, iron powder was removed by filtration, dichloromethane (100 mL) was added for extraction, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was evaporated to give the target product methyl 5-amino-4-fluoro-2-methoxybenzoate 064b (800 mg, white solid) in yield: 87 percent.
MS m/z(ESI)：200.1[M+1] + 。
Second step of
Preparation of methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate
Methyl 5-amino-4-fluoro-2-methoxybenzoate 064B (800mg, 2.23mmol) was dissolved in dichloromethane (30 mL), followed by addition of B deg.C-anhydride (4.38g, 20.08mmol), DMAP (49mg, 2.01mmol) and triethylamine (1.22mg, 12.05mmol) and stirring at 25 deg.C for 16h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and the solvent was removed. The crude product was concentrated after passing through a silica gel column (petroleum ether/ethyl acetate = 85/15) to give the desired product methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate 064c (300 mg, colorless oil) in yield: 22 percent.
MS m/z(ESI)：300.1[M+1] + 。
The third step
Preparation of N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamic acid tert-butyl ester
Methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate 064c (500mg, 1.67mmol) was dissolved in THF (10 mL) followed by the addition of diisobutylaluminum hydride (2.4 mL,1M in hexanes) at 0 deg.C. The reaction was stirred at room temperature for 2h. After completion of the reaction, saturated ammonium chloride was added to dissolve and quench, extraction was performed with ethyl acetate (3 × 20 mL), and after drying over anhydrous sodium sulfate, concentration was performed under reduced pressure to obtain a crude product, which was purified with a silica gel column (petroleum ether/ethyl acetate = 30%) and then concentrated to obtain N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamic acid tert-butyl ester 064d (230 mg, colorless oil), yield: 46 percent.
MS m/z(ESI)：294.2[M+1] + 。
The fourth step
Preparation of tert-butyl N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamate
N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamic acid tert-butyl ester 064d (230mg, 0.85mmol) was dissolved in dichloromethane (20 mL), followed by addition of carbon tetrabromide (337mg, 1.01mmol) and triphenylphosphine (267mg, 1.01mmol). The reaction solution was stirred at 25 ℃ for 16 hours. After completion of the reaction, concentration gave a crude product which was purified by silica gel column (petroleum ether/ethyl acetate = 7/3) and concentrated to give N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 064e (180 mg, white solid) in yield: 57 percent.
MS m/z(ESI)：356.2[M+1] + 。
The fifth step
Preparation of N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamic acid tert-butyl ester
Indole (105mg, 0.90mmol) was dissolved in tetrahydrofuran (10 mL) and NaH (36mg, 0.90mmol) was added. After the reaction was stirred at room temperature for 0.5h, N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 064e (150mg, 0.45mmol) was added. The reaction was stirred at room temperature for 16 hours. After completion of the reaction, sodium hydride was quenched by addition of ice water, followed by extraction with dichloromethane (3 × 20 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude product, which was purified by silica gel column (petroleum ether/ethyl acetate = 1/1) and then concentrated to remove the solvent to give tert-butyl N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamate 064f (60 mg, white solid) in yield: 34 percent.
MS m/z(ESI)：371.2[M+1] + 。
The sixth step
Preparation of 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline
N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamic acid tert-butyl ester 064f (60mg, 0.18mmol) was dissolved in dichloromethane (9 mL), followed by the addition of trifluoroacetic acid (3 mL). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction, concentration under reduced pressure gave 064g (40 mg, white solid) of 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline, yield: and 75 percent.
MS m/z(ESI)：271.1[M+1] + 。
Step seven
Preparation of 4- ({ [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
064g (40mg, 0.15mmol) of 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline was dissolved in tetrahydrofuran (20 mL), and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (70mg, 0.19mmol) and triethylamine (45mg, 0.44mmol) were added to stir the reaction mixture at 25 ℃ for 16 hours. Upon completion of the reaction monitored by LCMS, the compound 4- ({ [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 064h (60 mg, brown solid) was obtained in yield: and 55 percent.
MS m/z(ESI)：512.2[M+1] + 。
The eighth step
Preparation of 3- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction mixture in the seventh step. After addition of lithium hydroxide monohydrate (11mg, 0.25mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give the crude product, which was purified by preparative HPLC (0.1% trifluoroacetic acid/acetonitrile/water) to give 3- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2,4-dioxy-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester Cpd-008 (2 mg, white solid), yield: 4 percent.
MS m/z(ESI)：466.2[M+1] + 。
HPLC:98.19％(214nm),93.43％(254nm).
1 H NMR(400MHz,CD 3 OD)δ7.42(d,J＝8.0Hz,1H),7.27(d,J＝8.0Hz,1H),7.23(s,1H),7.03(t,J＝7.6Hz,2H),6.96(s,2H),6.93(dd,J＝9.2,6.0Hz,2H),4.04(s,2H),3.92(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-124.40--124.56.
Example 9
Preparation of 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-009)
Figure BDA0003608368340000351
First step of
Preparation of methyl 4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-hydroxybenzoate 065a (2g, 11.80mmol), methyl iodide (3g, 23.60mmol) and cesium carbonate (4g, 12.98mmol) were dissolved in acetonitrile (10 mL). The mixture was reacted at 25 ℃ for 6 hours, followed by extraction with ethyl acetate (3 × 50 mL), and the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain methyl 4-fluoro-2-methoxybenzoate 065b (2.1 g, clear oil), yield: 87 percent.
MS m/z(ESI)：185.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.84(dd,J＝8.8,6.4Hz,1H),6.69-6.64(m,2H),3.89(s,3H),3.87(s,3H).
Second step of
Preparation of methyl 5-bromo-4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-methoxybenzoate 065b (1.00g, 5.4 mmol), NBS (0.96g, 5.4 mmol), and ferric trichloride (90mg, 0.54mmol) were dissolved in acetonitrile (4 mL) and stirred at 60 ℃ for 1 hour. After completion of the reaction, the residue obtained after concentration was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain methyl 5-bromo-4-fluoro-2-methoxybenzoate 065c (1.10 g, white solid), yield: 68 percent.
MS m/z(ESI)：263.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ7.95(d,J＝8.0Hz,1H),7.31(d,J＝11.2Hz,1H),3.85(s,3H),3.78(s,3H).
The third step
Preparation of 5-bromo-4-fluoro-2-methoxybenzoic acid
Methyl 5-bromo-4-fluoro-2-methoxybenzoate 065c (1.00g, 3.80mmol) and lithium hydroxide (455mg, 19.00mmol) were dissolved in THF/H 2 O (10 mL). The mixture was stirred at 25 ℃ for 1 hour. After the reaction was completed, the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, followed by extraction with ethyl acetate (2X 30 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 5-bromo-4-fluoro-2-methoxybenzoic acid 065d (900 mg, white solid) in a yield of 76%.
MS m/z(ESI)：249.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ12.99(s,1H),7.92(d,J＝8.4Hz,1H),7.27(d,J＝11.2Hz,1H),3.84(s,3H).
The fourth step
Preparation of 5-bromo-4-fluoro-2-methoxybenzoyl chloride
5-bromo-4-fluoro-2-methoxybenzoic acid 065d (900mg, 3.6 mmol) was dissolved in anhydrous dichloromethane (20 mL), and oxalyl chloride (3.00g, 24mmol) and DMF (2.00g, 28mmol) were added dropwise. The mixture was stirred at 25 ℃ for 0.5 hour. After the reaction was completed, the reaction mixture was concentrated to remove excess solvent and oxalyl chloride, and the obtained 5-bromo-4-fluoro-2-methoxybenzoyl chloride 065e (900 mg, white solid) was used directly in the next reaction with a yield of 65%.
MS m/z(ESI)：263.0(M-Cl+CH 3 )。
The fifth step
Preparation of 1- [ (5-bromo-4-fluoro-2-methoxyphenyl) carbonyl ] indole
1H-indole (2g, 20.30mmol) was dissolved in tetrahydrofuran (10 mL). Sodium hydride (1.00g, 27.20mol) was then added at 0 ℃. The mixture was stirred at 25 ℃ for 1 hour, and then 5-bromo-4-fluoro-2-methoxybenzoyl chloride 065e (900mg, 3.4 mmol) was dissolved in tetrahydrofuran (5 mL) and added dropwise to the solution, with continued stirring at 25 ℃. After 1.5 h, the reaction was quenched by slowly pouring it into ice water and adjusting its pH to 6-7 with 1M dilute hydrochloric acid, extracted with dichloromethane (2X 30 mL), the organic phases combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 3/7) to give 1- [ (5-bromo-4-fluoro-2-methoxyphenyl) carbonyl ] indole 065f (400 mg, white solid) in 27% yield.
MS m/z(ESI)：348.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ8.29(d,J＝8.0Hz,1H),7.95(d,J＝8.0Hz,1H),7.64(d,J＝7.2Hz,1H),7.39-7.30(m,3H),7.22(d,J＝3.6Hz,1H),6.70(d,J＝3.6Hz,1H),3.77(s,3H).
The sixth step
Preparation of N- (diphenylmethylene) -2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline
1- [ (5-bromo-4-fluoro-2-methoxyphenyl) carbonyl ] indole 065f (320mg, 0.92mmol), benzhydrylamine (666mg, 3.68mmol), BINAP (114mg, 0.18mmol), tris (dibenzylideneacetone) dipalladium (84mg, 0.09mmol) and cesium carbonate (599mg, 1.84mmol) were dissolved in DMF (5 mL). The mixture was reacted at 120 ℃ for 6 hours. After the reaction was completed, quenched with water (50 mL), and extracted with dichloromethane (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 065g (220 mg, yellow oil) of N- (diphenylmethylene) -2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline, yield: and 43 percent.
MS m/z(ESI)：449.1[M+1] + 。
Seventh step
Preparation of 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline
065g (110mg, 0.24mmol) of N- (diphenylmethylene) -2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline was dissolved in tetrahydrofuran (5 mL), concentrated hydrochloric acid (1 mL) was slowly added dropwise, and the mixture was reacted at 25 ℃ for 5 minutes. After the reaction was completed, the pH was adjusted to 7 to 8 with a saturated sodium bicarbonate solution and extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried with anhydrous sodium sulfate, and the obtained residue was purified with a silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline 065h (80 mg, orange oil), yield: 74 percent.
MS m/z(ESI)：285.1[M+1] + 。
Eighth step
Preparation of dimethyl 4- [ ({ { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline 065h (40mg, 0.14mmol) and methyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (236 mg, 0.70mmol) were dissolved in tetrahydrofuran (5 mL) and then triethylamine (79mg, 1.41mmol) was added, and the mixture was stirred at 25 ℃ for 16 hours after completion of the reaction, the residue obtained after concentration was purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to give 4- [ ({ { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester preparation 065i (50 mg, white solid) with a yield of 34%.
MS m/z(ESI)：526.0[M+1] + 。
The ninth step
Preparation of methyl 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylate
Preparation of methyl 4- [ ({ { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 065i (50mg, 0.09mmol) was dissolved in tetrahydrofuran (5 mL), and lithium hydroxide (11mg, 0.47mmol) was dissolved in water (5 mL) and added to the above solution. The mixture was reacted at 25 ℃ for 5 minutes. After the reaction was complete, it was extracted with dichloromethane (3X 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated to give methyl 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylate 065j (35 mg, yellow solid) in yield: 44 percent.
MS m/z(ESI)：494.0[M+1] + 。
The tenth step
Preparation of 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methyl 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylate 065j (35mg, 0.0709 mmol) and trimethyltin hydroxide (64mg, 0.3545mmol) were dissolved in 1,2-dichloroethane (4 mL). The mixture was reacted at 80 ℃ for 16 hours. After the reaction was completed, the reaction mixture was directly concentrated and preliminarily purified by reverse column (water: acetonitrile = 3:2) to prepare 3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-009 (2.31 mg, white solid) with a yield of 6%.
MS m/z(ESI)：480.0[M+1] + 。
HPLC:100％(214nm),99.36％(254nm)。
1 H(400MHz,d 6 -DMSO)δ12.02(s,1H),8.27(d,J＝8.0Hz,1H),7.76(d,J＝8.3Hz,1H),7.66(d,J＝7.2Hz,1H),7.44(d,J＝12.0Hz,1H),7.39-7.30(m,3H),7.20(d,J＝3.6Hz,1H),6.76(d,J＝3.6Hz,1H),3.84(s,3H).
Example 10
Preparation of 3- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-010)
Figure BDA0003608368340000371
First step of
Preparation of 2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione
2-fluoro-4-methoxyaniline 66a (500mg, 3.54mmol) was dissolved in N, N-dimethylformamide (10 mL) and phthalic anhydride (577 mg, 3.89mmol) was added. The reaction solution was stirred at 120 ℃ for 4 hours. After cooling to room temperature, the dichloromethane was extracted three times, and the dichloromethane layer was washed with a saturated NaCl solution and then with anhydrous Na 2 SO 4 Drying, filtering, and concentrating under reduced pressure to obtain 2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione 66b (500 mg, white solid), yield: 49 percent.
MS m/z(ESI)：272.2[M+1] + 。
Second step of
Preparation of 5- (1,3-dioxoisopropan-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione 66b (200mg, 0.74mmol) was dissolved in chlorosulfonic acid (3.44mL, 29.49mmol) and stirred at room temperature for 2h. After completion of the reaction, the reaction solution was poured into ice water, and the aqueous phase was extracted with ethyl acetate (2X 30 mL) and the organic phase was discarded. The remaining aqueous phase was adjusted to pH 5 with 5N aqueous hydrochloric acid and extracted with ethyl acetate (3X 30 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 5- (1,3-dioxoisopropyl-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 66c (250 mg, white solid) yield: and 64 percent.
MS m/z(ESI)：392.0[M+1] + 。
The third step
Preparation of 2- { [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid
Indole (95mg, 0.81mmol) was dissolved in THF (5 mL), naH (95mg, 0.81mmol, kerosene-preserved, content 60%) was added to the reaction solution while cooling on ice, and after stirring at room temperature for 0.5h, 5- (1,3-dioxoisopropanol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 66c (150mg, 0.41mmol) was added. The reaction mixture was stirred at room temperature for 4h, after completion of the reaction, water was added, dichloromethane was extracted three times, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to give 66d 2- { [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid (180 mg, white solid) in yield: 85 percent.
MS m/z(ESI)：469.1[M+1] + 。
The fourth step
Preparation of 2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione
2- { [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl]Carbamoyl } benzoic acid 66d (130mg, 0.32mmol) and sodium acetate (79mg, 0.96mmol) were dissolved in acetic anhydride (5 mL). The reaction solution is in 9Stirred at 0 ℃ for 2 hours. After the reaction was complete, the reaction was cooled to room temperature and then saturated NaHCO was used 3 Wash and extract with ethyl acetate (3X 30 mL). The organic phases are combined, washed with saturated salt water, dried by anhydrous sodium sulfate and concentrated to obtain 2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ]Isoindole-1,3-dione 66e (130 mg, colorless gum), yield: 81 percent.
MS m/z(ESI)：451.1[M+1] + 。
The fifth step
Preparation of 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyaniline
2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione 66e (140mg, 0.31mmol) was dissolved in ethanol (10 mL) and then hydrazine hydrate (1.56mL, 31.08mmol) was added and stirred at 90 ℃ for 1h. After completion of the reaction, the solvent was evaporated and then extracted with dichloromethane (3X 20 mL). The organic phases were combined, washed with brine and dried over anhydrous sodium sulfate. The residue obtained after concentration was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to give 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyaniline 66f (80 mg, white solid) in yield: and 76 percent.
MS m/z(ESI)：321.1[M+1] + 。
The sixth step
Preparation of 4- ({ [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-5- (indole-1-sulfonyl) -4-methoxyaniline 66f (50mg, 0.16mmol) was dissolved in anhydrous tetrahydrofuran (9 mL), followed by the addition of triethylamine (47mg, 0.47mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (73mg, 0.20mmol). The reaction was stirred at room temperature for 16h. After the reaction is finished, the next reaction is directly carried out without post-treatment.
MS m/z(ESI)：562.1[M+1] + 。
Seventh step
Preparation of 3- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
MeOH (1 mL) and water (1 mL) were added to 66g of the reaction solution in the sixth step, followed by lithium hydroxide monohydrate (11mg, 0.25mmol). The reaction was stirred at room temperature for 2h. After completion of the reaction the solvent was evaporated to give the crude product, which was purified by preparative HPLC (0.1% formic acid/acetonitrile/water) to give 3- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-010 (20 mg, white solid) in yield: 46 percent.
MS m/z(ESI)：516.0[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,CD 3 OD)δ8.38(d,J＝8.0Hz,1H),7.70(d,J＝6.8Hz,1H),7.65(d,J＝3.6Hz,1H),7.58-7.53(m,1H),7.34(s,1H),7.22(dd,J＝9.2,5.6Hz,2H),7.12(d,J＝11.2Hz,1H),6.69(d,J＝3.6Hz,1H),3.74(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-108.72.
Example 11
Preparation of 3- (5- (((1H-indol-3-yl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-011)
Figure BDA0003608368340000391
First step of
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxybenzene
4-fluoro-2-methoxyphenol (1.00g, 7.04mmol) and K 2 CO 3 (1.5g, 10.55mmol) was dissolved in acetone (10 mL) and benzyl bromide (1.4g, 8.4mmol) was added. Reacting for 16h at 65 ℃. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE: etOAc =50 1) to afford preparation 068b (1.4 g, white solid) of 1- (benzyloxy) -4-fluoro-2-methoxybenzene, yield: 77 percent.
MS m/z(ESI)：233.2[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ7.42(d,J＝7.2Hz,2H),7.39-7.33(m,2H),7.33-7.28(m,1H),6.80(dd,J＝8.8,5.6Hz,1H),6.65(dd,J＝10.2,3.0Hz,1H),6.55-6.49(m,1H),5.09(s,2H),3.86(s,3H).
Second step of
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene
Compound 068b (500mg, 2.15mmol) was dissolved in acetic anhydride (3 mL) and nitric acid (65%) (0.2 mL) was slowly added dropwise at 0 ℃. The reaction is carried out for 4h at 0 ℃. The reaction solution was quenched with water (10 mL), extracted with dichloromethane (3X 20 mL), washed with saturated NaCl water, and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE: etOAc =10 1) to give 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene 068c (430 mg, light yellow solid) in yield: 58 percent.
MS m/z(ESI)：278.1[M+1] + .
The third step
Preparation of 4-fluoro-2-methoxy-5-nitrophenol
The compound 068c (550mg, 1.98mmol) was dissolved in a solution of boron trichloride in methylene chloride (1M, 5mL) and reacted at room temperature for 2 hours. The reaction was quenched with water (10 mL), extracted with dichloromethane DCM (3X 10 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE: etOAc = 2:1) to give 4-fluoro-2-methoxy-5-nitrophenol 068d (200 mg, yellow solid) in yield: 50 percent.
1 H NMR(400MHz,CDCl 3 )δ7.66(d,J＝7.2Hz,1H),6.74(d,J＝11.8Hz,1H),5.56(s,1H),4.00(s,3H).
The fourth step
Preparation of 3- (4-fluoro-2-methoxy-5-nitrophenoxy) -1-tolyl-1H-indole
(2S, 3R) -N, N, N-triethyl-2-hydroxy-1- (4-methyl-1-sulfonylphenyl) -2,3-indoline-3-ammonium bromide (500mg, 1mmol), compound 068d (994.6mg, 5mmol) and triethylamine (537.8mg, 5mmol) were dissolved in ethyl acetate (30 mL) and reacted at 80 ℃ for 5h. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (3X 30 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting intermediate was dissolved in ethyl acetate (30 mL), and boron trifluoride diethyl etherate (750 mg, 5mmol) was added. React for 3h at 50 ℃. Reacting the reaction solution With saturated NaHCO 3 The solution was quenched (50 mL), extracted with ethyl acetate (3X 30 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE: etOAc = 1:1) to give 068e (350 mg, light brown solid), yield: and 72 percent.
MS m/z(ESI)：457.1[M+1] + .
The fifth step
Preparation of 2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) aniline
Compound 068e (700mg, 1.53mmol) is dissolved in methanol (15 mL) and saturated NH 4 To a mixed solution of Cl (5 mL) solution was then added iron powder (857mg, 15.3mmol). The reaction was carried out at room temperature for 16h. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was dissolved in ethyl acetate (20 mL) and saturated NaHCO was used 3 And NaCl solution, and drying the product by anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) aniline 068f (470 mg, brown solid) in yield: 65 percent.
MS m/z(ESI)：427.2[M+1] + .
The sixth step
Preparation of methyl 4- (3- (2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylate
Compound 068f (420mg, 0.98mmol), triethylamine (298mg, 2.95mmol) and 2,3-dimethyl-4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (395mg, 1.18mmo) were dissolved in THF (20 mL) and reacted at room temperature for 16h. The reaction was concentrated under reduced pressure to give 068g (600 mg, brown oily liquid) of 4- (3- (2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester in yield: and 55 percent.
MS m/z(ESI)：668.1[M+1] + .
Seventh step
Preparation of 3- (2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
068g (600mg, 0.90mmol) of compound and monohydrate and lithium hydroxide (188mg, 4.49mmol) were dissolved in THF/MeOH/H 2 O =5, 1 (20 mL), and reacted at room temperature for 3h. Concentrating the reaction solution under reduced pressure, and subjecting the residue to reverse phase column chromatography (ACN: H) 2 O (0.2% FA) = 60%) purification to give 3- (2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d]Pyrimidine-5-carboxylic acid 068h (200 mg, brown solid), yield: 32 percent.
MS m/z(ESI)：622.1[M+1] + .
The eighth step
Preparation of 3- (5- (((1H-indol-3-yl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 068h (100mg, 0.16mmol) was dissolved in a mixed solution of THF/MeOH =2:1 (10 mL), and cesium carbonate (262mg, 0.80mmol) was added and reacted at room temperature for 16h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to preparative HPLC (ACN: H) 2 O(0.1％NH 3 ) = 95%) and purifying to obtain 3- (5- (((1H-indole-3-yl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d]Pyrimidine-5-carboxylic acid Cpd-011 (10 mg, light yellow solid), yield: 13 percent.
MS m/z(ESI)：468.1[M+1] + .
1 H NMR(400MHz,DMSO-d 6 )δ10.91(s,1H),7.32(t,J＝8.4Hz,2H),7.15(d,J＝10.2Hz,2H),7.07(q,J＝7.2Hz,1H),7.00-6.95(m,1H),6.74(d,J＝7.6Hz,1H),6.28-6.26(m,1H),3.91(s,3H).
19 F NMR(376MHz,DMSO-d 6 )δ-126.46(s,1H).
Example 12
Preparation of 3- (5- ((1H-indol-3-yl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-012)
Figure BDA0003608368340000411
First step of
Preparation of 1- (3- ((4-fluoro-2-methoxy-5-nitrophenyl) amino) -1H-indol-1-yl) ethan-1-one
To a solution of 1-acetyl-2H-indol-3-one 069a (1.5g, 8.6 mmol) in AcOH (15 mL) was added 4-fluoro-2-methoxy-5-nitroaniline (1.6 g,8.6 mmol). The reaction mixture was stirred at 120 ℃ for 8 hours. The reaction was filtered, the cake was slurried with methanol and dried to give the product 1- (3- ((4-fluoro-2-methoxy-5-nitrophenyl) amino) -1H-indol-1-yl) ethan-1-one 069b (0.8 g, red solid). The yield is 24%.
MS m/z(ESI)：344.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ8.39(d,J＝8.4Hz,1H),7.72(s,1H),7.63(s,1H),7.47(d,J＝7.6Hz,1H),7.38(t,J＝7.2Hz,1H),7.33-7.22(m,3H),4.05(s,3H),2.62(s,3H).
Second step of
Preparation of 1- {3- [ (5-amino-4-fluoro-2-methoxyphenyl) amino ] indol-1-yl } ethanone
To a solution of 1- {3- [ (4-fluoro-2-methoxy-5-nitrophenyl) amino ] indol-1-yl } ethanone 069b (200mg, 0.58mmol) in ethyl acetate/dichloromethane (20 mL/1 mL) was added Pd/C (25 mg). The reaction mixture was stirred at 25 ℃ for 8 hours. The reaction was filtered and concentrated to give the product 1- {3- [ (5-amino-4-fluoro-2-methoxyphenyl) amino ] indol-1-yl } ethanone 069c (70 mg, brown solid). The yield was 26.8%.
MS m/z(ESI)：314.2[M+1] + 。
The third step
Preparation of dimethyl 4- { [ (2-fluoro-4-methoxy-5- { [1- (4-methyl-1-sulfonylphenyl) indol-3-yl ] amino } phenyl) carbamoyl ] amino } thiophene-2,3-dicarboxylate
To a solution of 1- {3- [ (5-amino-4-fluoro-2-methoxyphenyl) amino ] indol-1-yl ] ethanone 069c (70mg, 0.22mmol) in THF (3 mL) were added triethylamine (226mg, 2.2mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (187mg, 0.56mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was concentrated. The concentrated residue was purified on a flash preparative column to give the product dimethyl 4- { [ (2-fluoro-4-methoxy-5- { [1- (4-methyl-1-sulfonylphenyl) indol-3-yl ] amino } phenyl) carbamoyl ] amino } thiophene-2,3-dicarboxylate 069d (110 mg, a red brown solid). The yield was 62%.
MS m/z(ESI)：555.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.78(s,1H),8.51(s,1H),7.88(s,1H),7.81(d,J＝7.6Hz,1H),7.53(d,J＝10.4Hz,2H),7.43-7.36(m,1H),7.30(d,J＝7.6
Hz,1H),6.73(d,J＝11.7Hz,1H),6.49(s,1H),3.93-3.86(t,J＝12Hz,9H),2.64(s,3H).
The fourth step
Preparation of 3- (5- ((1H-indol-3-yl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To 4- { [ (2-fluoro-4-methoxy-5- { [1- (4-methyl-1-sulfonylphenyl) indol-3-yl group]Amino } phenyl) carbamoyl]Amino } thiophene-2,3-dicarboxylic acid dimethyl ester 069d (110mg, 0.20mmol) in THF/MeOH/H 2 To a solution of O = 1. The reaction mixture was stirred at 25 ℃ for 20 minutes. And purifying the reaction solution by a quick column to obtain a crude product. Purifying the crude product by high performance liquid chromatography to obtain the product 3- (5- ((1H-indole-3-yl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ]Pyrimidine-5-carboxylic acid Cpd-012 (1.02 mg, brown solid). Yield: 2.33 percent.
MS m/z(ESI)：467.0[M+1] + 。
1 H NMR(400MHz,CD 3 OD)δ7.31(dd,J＝23.6,8.0Hz,2H),7.14(s,1H),7.05(t,J＝7.2Hz,2H),6.91(dd,J＝19.4,11.6Hz,2H),6.44(d,J＝7.6Hz,1H),3.99(s,3H).
Example 13
Preparation of 3- [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-013)
Figure BDA0003608368340000421
First step of
Preparation of 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene
Under a nitrogen atmosphere, tert-butyl nitrite (8.32g, 80.7 mmol) and copper bromide (9.01, 40.3mmol) were dissolved in acetonitrile (40 mL), the reaction solution was heated to 50 ℃ and then a solution of 4-fluoro-2-methoxy-5-nitroaniline 070a (5g, 26.9 mmol) in acetonitrile (20 mL) was added dropwise, and the reaction solution was stirred at 50 ℃ for 3 hours. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate = 10/1) to give the title product, 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 070b (5.2 g, yellow solid), 73% yield.
MS m/z(ESI)：250.0[M+1] + .
1 H NMR(400MHz,DMSO-d 6 )δ8.41(d,J＝8.4Hz,1H),7.44(d,J＝13.6Hz,1H),4.00(s,3H).
Second step of
Preparation of 5-bromo-2-fluoro-4-methoxyaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 070b (2g, 8mmol) was dissolved in ethanol/water =4/1 (20 mL), and iron powder (2.23g, 40mmol) and ammonium chloride (2.14g, 40mmol) were added. The reaction solution was stirred at 90 ℃ for 2 hours. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate = 3/2) to give the title product 5-bromo-2-fluoro-4-methoxyaniline 070c (1.5 g, brown solid) with a yield of 81%.
MS m/z(ESI)：220.1[M+1] + .
The third step
Preparation of tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate
5-bromo-2-fluoro-4-methoxyaniline 070c (1.5g, 6.8mmol) was dissolved in tetrahydrofuran (20 mL), di-tert-butyl dicarbonate (1.78g, 8.1mmol) and triethylamine (2.06g, 20.4 mmol) were added, and the reaction mixture was stirred at 60 ℃ for 16 hours. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain the title product (5-bromo-2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester 070d (1.3 g, yellow solid) with a yield of 56%.
MS m/z(ESI)：342.0[M+1] + .
The fourth step
Preparation of tert-butyl [ 2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl ] carbamate
Under a nitrogen atmosphere, (5-bromo-2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester 070d (1.2g, 3.7 mmol) was dissolved in dioxane (15 mL), 4,4,5,5-tetramethyl-2- (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) -1,3,2-dioxaborane (1.41g, 5.5 mmol), 1,1' -bis (diphenylphosphino) ferrocene palladium (0.27g, 0.3 mmol), and potassium acetate (0.73g, 7.4 mmol) were added, and the reaction was stirred at 90 ℃ for 8 hours. The residue obtained by concentrating the reaction liquid was purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to obtain the title product [ tert-butyl 2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl ] carbamate 070e (1.1 g, yellow solid) in a yield of 75%.
MS m/z(ESI)：390.2(M+23).
The fifth step
Preparation of tert-butyl [5- (dihydroxyboranyl) -2-fluoro-4-methoxyphenyl ] carbamate
[ 2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) phenyl ] carbamic acid tert-butyl ester 070e (1.1g, 3mmol) was dissolved in tetrahydrofuran (20 mL) and water (10 mL), sodium periodate (1.93g, 9mmol) and ammonium chloride (0.48g, 9mmol) were added, and the reaction was stirred at 25 ℃ for 16 hours. Water (100 mL) was added to the reaction solution, followed by extraction with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by concentration was purified by a silica gel column (petroleum ether/ethyl acetate = 9/1) to obtain the title product [5- (dihydroxyboranyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 070f (0.55 g, yellow solid) with a yield of 56%.
MS m/z(ESI)：308.2(M+23).
1 H NMR(400MHz,DMSO-d 6 )δ8.63(s,1H),7.68(s,2H),7.56(d,J＝9.6Hz,1H),6.88(d,J＝12.8Hz,1H),3.78(s,3H),1.43(s,9H).
The sixth step
Preparation of {5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamic acid tert-butyl ester
1-benzofuran-3-carbaldehyde (256mg, 1.75mmol) was dissolved in toluene (5 mL) under a nitrogen atmosphere, and [5- (dihydroxyboryl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 070f (500mg, 1.75mmol), 2- [ bis (2,4-di-tert-butyl-phenoxy) phosphinoxy ] -3,5-di (tert-butyl) phenyl-palladium (II) chloride dimer (138mg, 0.09mmol), and potassium phosphate (742mg, 3.5 mmol) were added, and the reaction solution was stirred at 50 ℃ for 6 hours. The residue obtained by the concentration was purified by a silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 070g (40 g, colorless oil) of tert-butyl {5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamate as the title product, yield: 6%.
MS m/z(ESI)：410.1(M+23).
Seventh step
Preparation of tert-butyl [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamate
[5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamic acid tert-butyl ester 070g (40mg, 0.11mmol) was dissolved in dichloromethane (2 mL) under a nitrogen atmosphere, triethylsilane (24mg, 0.21mmol) was added under an ice-water bath, and after stirring for 10 minutes, trifluoroacetic acid (36mg, 0.31mmol) was added, and the reaction mixture was stirred at 25 ℃ for 1 hour. Water (50 mL) was added to the reaction mixture, which was then extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to give the title product tert-butyl [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamate 070h (35 mg, yellow oil) in 82% yield.
MS m/z(ESI)：394.1(M+23).
Eighth step
Preparation of 5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyaniline
Tert-butyl [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamate 070h (35mg, 0.1mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (215mg, 1.88mmol) was added, and the reaction was stirred at 25 ℃ for 1 hour. The residue obtained by the concentration was purified by a silica gel column (petroleum ether/ethyl acetate = 3/2) to obtain the title product 5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyaniline 070i (25 mg, yellow oil) in a yield of 78%.
MS m/z(ESI)：272.2[M+1] + .
The ninth step
Preparation of dimethyl 4- ({ [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyaniline 070i (30mg, 0.11mmol) was dissolved in tetrahydrofuran (3 mL), 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (37mg, 0.111mmol) and triethylamine (34mg, 0.33mmol) were added in this order, the reaction was stirred at 25 ℃ for 16 hours, and the reaction was concentrated to give the title product 4- ({ [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 070j (40 mg, yellow oil) in 56% yield.
MS m/z(ESI)：513.0[M+1] + .
The tenth step
Preparation of 3- [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 070j (40mg, 0.08mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (17mg, 0.39mmol) was added, and the reaction mixture was stirred at 25 ℃ for 2 hours. Water (0.5 mL) was then added and the reaction was stirred at 25 ℃ for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution and then extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by preparative HPLC (acetonitrile/water (0.1% formic acid) to give the title product 3- [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-013 (5 mg, white solid) in 13% yield.
MS m/z(ESI)：466.9[M+1] + .
HPLC:98.37％(214nm),97.95％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.54(s,1H),11.90(s,1H),7.75(s,1H),7.54(dd,J＝9.6,8.8Hz,2H),7.34-7.26(m,3H),7.22-7.17(m,1H),7.13(d,J＝12.0Hz,1H),3.95(s,2H),3.92(s,3H).
Example 14
Preparation of 3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-014)
Figure BDA0003608368340000451
First step of
Preparation of 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene
Cuprous bromide (5.79g, 0.04mol) and tert-butyl nitrite (8.32g, 0.08mol) were dissolved in acetonitrile (20 mL), stirred at 50 ℃ for 10 minutes, a solution of 4-fluoro-2-methoxy-5-nitroaniline (5.00g, 0.03mol) in acetonitrile (30 mL) was added to the reaction solution, and the reaction solution was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 9/1) to obtain 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 071b (4.9 g, yellow solid) in 70% yield.
MS m/z(ESI)：250.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.36(d,J＝8.0Hz,1H),6.78(d,J＝12.4Hz,1H),4.01(s,3H)。
Second step of
Preparation of 5-bromo-2-fluoro-4-methoxyaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 071b (4.88g, 0.02mol), iron powder (2.18g, 0.04mmol) and ammonium chloride (2.09g, 0.04mmol) were dissolved in ethanol: water =1:1 (20 mL), and the reaction mixture was reacted at 25 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 7/3) to obtain 5-bromo-2-fluoro-4-methoxyaniline 071c (2.11 g, yellow solid) in 48% yield.
MS m/z(ESI)：220.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.00(d,J＝9.4Hz,1H),6.66(d,J＝12.2Hz,1H),3.80(s,3H)。
The third step
Preparation of tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate
5-bromo-2-fluoro-4-methoxyaniline 071c (2g, 9.1mmol) and di-tert-butyl dicarbonate (2.98g, 13.6 mmol) were dissolved in tetrahydrofuran (20 mL), triethylamine (1.84g, 18.2mmol) was added at room temperature, and the reaction mixture was reacted at 60 ℃ for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was purified by silica gel column (petroleum ether/ethyl acetate = 15/1) to obtain tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate 071d (788 mg, yellow solid) in 26% yield.
MS m/z(ESI)：264.0(M-56)。
1 H NMR(400MHz,CDCl 3 )δ8.20(s,1H),6.61(d,J＝12.4Hz,1H),6.40(s,1H),3.77(s,3H),1.45(s,9H)。
The fourth step
Preparation of tert-butyl (2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl) carbamate
Tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate 071d (438mg, 1.36mmol), 4,4,5,5-tetramethyl-2- (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) -1,3,2-dioxaborane (415mg, 1.36mmol), 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium (100mg, 0.14mmol) and potassium acetate (267mg, 2.7mmol) were dissolved in 1,4-dioxane (10 mL) and the reaction solution was reacted at 90 ℃ for 10 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). The obtained residue was purified by silica gel column (petroleum ether/ethyl acetate = 9/1) to obtain (2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) phenyl) carbamic acid tert-butyl ester 071e (358 mg, yellow solid) in 70% yield.
MS m/z(ESI)：312.1(M-56)。
1 H NMR(400MHz,CDCl 3 )δ8.13(s,1H),6.61(d,J＝13.0Hz,1H),6.33(s,1H),3.78(s,3H),1.51(s,6H),1.34(s,9H),1.27(s,3H),1.24(s,3H)。
The fifth step
Preparation of (5- ((tert-butoxycarbonyl) amino) -4-fluoro-2-methoxyphenyl) boronic acid
Tert-butyl (2-fluoro-4-methoxy-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl) phenyl) carbamate 071e (345mg, 0.93mmol), sodium periodate (601mg, 2.8mmol) and ammonium chloride (150mg, 0.08mmol) were dissolved in tetrahydrofuran: water =3:1 (20 mL), and the reaction mixture was reacted at room temperature for 48 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give (5- ((tert-butoxycarbonyl) amino) -4-fluoro-2-methoxyphenyl) boronic acid 071f (212 mg, grey solid) in 73% yield.
MS m/z(ESI)：230.1(M-56)。
The sixth step
Preparation of tert-butyl (5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamate
(5- ((tert-butoxycarbonyl) amino) -4-fluoro-2-methoxyphenyl) boronic acid 071f (200mg, 0.7 mmol), 1-benzothiophene-3-carbaldehyde (170mg, 1.05mmol), 2- [ bis (2,4-di-tert-butyl-phenoxy) phosphinyloxy ] -3,5-di (tert-butyl) phenyl-palladium (II) chloride dimer (55mg, 0.03mmol), and potassium phosphate (296mg, 1.39mmol) were dissolved in toluene (10 mL), and the reaction solution was reacted at 50 ℃ for 2 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. Obtained residue the obtained residue was purified by silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain 071g (75 mg, yellow solid) of (tert-butyl 5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamate, a yield of 25%.
MS m/z(ESI)：386.2(M-17)。
1 H NMR(400MHz,CDCl 3 )δ7.89(d,J＝5.8Hz,1H),7.86-7.80(m,1H),7.40-7.29(m,2H),7.21(s,1H),6.71(d,J＝12.2Hz,1H),6.36(s,1H),3.79(s,2H),1.46(s,7H)。
Seventh step
Preparation of 5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyaniline
071g (65mg, 0.13mmol) of t-butyl (5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamate, triethylsilane (19mg, 0.16mmol) and trifluoroacetic acid (45mg, 0.34mmol) were dissolved in dichloromethane (10 mL), and the reaction solution was reacted at room temperature for 10 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with dichloromethane (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. Obtained residue the obtained residue was purified by silica gel column (petroleum ether/ethyl acetate = 4/1) to give 5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyaniline 071h (47 mg, grey solid) in 96% yield.
MS m/z(ESI)：288.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.90-7.79(m,1H),7.75-7.64(m,1H),7.41-7.30(m,2H),6.98(s,1H),6.65(d,J＝12.4Hz,1H),6.54(d,J＝9.8Hz,1H),4.07(s,2H),3.77(s,3H)。
Eighth step
Preparation of methyl 4- (3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyaniline 071h (10mg, 0.14mmol) and 2,3-dimethyl-4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (56mg, 0.11116mmol) were dissolved in tetrahydrofuran (8 mL), triethylamine (28mg, 0.28mmol) was added at room temperature, the reaction liquid was reacted at room temperature for 16 hours, after the reaction was completed, water (20 mL) was added to the reaction liquid, and extraction was performed with ethyl acetate (3 × 20 mL), all organic phases were combined, anhydrous sodium sulfate was dried and concentrated, the resultant residue was purified with a silica gel column (petroleum ether/ethyl acetate = 7/3) to give (4- (3- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid methyl ester (31 mg, 07140% yield as a yellow solid.
MS m/z(ESI)：529.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.85(s,1H),8.74(s,1H),7.96(dd,J＝6.6,2.2Hz,1H),7.85(s,1H),7.79(dd,J＝6.6,2.2Hz,1H),7.70(d,J＝9.4Hz,1H),7.41-7.32(m,2H),7.26(s,1H),7.03(d,J＝13.0Hz,1H),4.08(s,2H),3.86(s,3H),3.81(s,6H)。
The ninth step
Preparation of 3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 071i (25mg, 0.05mmol) and lithium hydroxide (11mg, 0.5mmol) were dissolved in tetrahydrofuran: methanol: water =1:1:1 (6 mL), and the reaction mixture was reacted at room temperature for 1 hour. After the reaction was completed, the reaction solution was quenched to pH <7 by adding 1N hydrochloric acid, and extracted with ethyl acetate (3X 10 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified using a reverse phase column (47% acetonitrile/water) to afford 3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid, cpd-014 (7.92 mg, white solid) in 34% yield.
MS m/z(ESI)：483.0[M+1] + 。
HPLC:99.67％(214nm),98.99％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ7.96(dd,J＝6.6,2.2Hz,1H),7.84(dd,J＝6.6,2.2Hz,1H),7.41-7.33(m,2H),7.26(s,1H),7.20(d,J＝8.6Hz,1H),7.12(d,J＝12.0Hz,1H),6.95(s,1H),4.11(s,2H),3.89(s,3H)。
Example 15
Preparation of 3- { 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-015)
Figure BDA0003608368340000471
First step of
Preparation of methyl 4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-hydroxybenzoate 072a (3.00g, 17.63mmol) was dissolved in acetonitrile (30 mL) and iodomethane (3.75g, 26.45mmol) and cesium carbonate (11.49g, 35.26mmol) were added. The reaction solution was stirred at room temperature for 16 hours. Upon completion of the reaction by LCMS, extraction with dichloromethane (3 × 100 mL) was added, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and then subjected to silica gel column (petroleum ether/ethyl acetate = 6/4) after evaporation of the solvent to obtain the objective product methyl 4-fluoro-2-methoxybenzoate 072b (3.00 g, anhydrous oil), yield: 83 percent.
MS m/z(ESI)：185.2[M+1] + 。
Second step of
Preparation of methyl 4-fluoro-2-methoxy-5-nitrobenzoate
Methyl 4-fluoro-2-methoxybenzoate 072b (1.00g, 5.43mmol) was dissolved in concentrated sulfuric acid (4 mL), followed by addition of concentrated nitric acid (0.5 mL) while cooling on ice. Stirring at 0 deg.C for 0.5h. After completion of the reaction, the reaction solution was poured into ice, the pH was adjusted to 7 to 8 by addition of a saturated sodium carbonate solution, followed by extraction with methylene chloride (3X 30 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and then the solvent was removed. The crude product was passed through a silica gel column (petroleum ether/ethyl acetate = 30/70) and concentrated to give the target product methyl 4-fluoro-2-methoxy-5-nitrobenzoate 072c (700 mg, white solid) in yield: 53 percent.
MS m/z(ESI)：230.2[M+1] + 。
The third step
Preparation of 4-fluoro-2-methoxy-5-nitrobenzoic acid
Methyl 4-fluoro-2-methoxy-5-nitrobenzoate 072c (300mg, 1.75mmol) was dissolved in THF (10 mL) and water (2 mL) followed by addition of sodium hydroxide (262mg, 5.23mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction, the pH was adjusted to 4-5 by addition of 1N hydrochloric acid, and the mixture was extracted with dichloromethane (3X 20 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4-fluoro-2-methoxy-5-nitrobenzoic acid 072d (300 mg, white solid) in yield: 72 percent.
MS m/z(ESI)：216.2[M+1] + 。
The fourth step
Preparation of 4-fluoro-2-methoxy-5-nitrobenzoyl chloride
4-fluoro-2-methoxy-5-nitrobenzoic acid 072d (100mg, 0.46mmol) was dissolved in dichloromethane (10 mL) followed by addition of oxalyl chloride (118mg, 0.93mmol) and DMF (0.1 mL). The reaction solution was stirred at 25 ℃ for 1 hour. After completion of the reaction, concentration was carried out to give 4-fluoro-2-methoxy-5-nitrobenzoyl chloride 072e (120 mg, white solid), yield: 88 percent.
MS m/z(ESI)：230.2(M-Cl+CH 3 )。
The fifth step
Preparation of 3- [ (4-fluoro-2-methoxy-5-nitrophenyl) carbonyl ] -1H-indole
4-fluoro-2-methoxy-5-nitrobenzoyl chloride 072e (120mg, 0.51mmol) and aluminum trichloride (137mg, 1.02mmol) were dissolved in dichloromethane (15 mL) followed by the addition of indole (120mg, 1.02mmol). The reaction was stirred at room temperature for 1 h. After completion of the reaction, water (10 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate, the solvent was evaporated and concentrated after filtration to give a crude product, which was concentrated off after passing through a silica gel column (petroleum ether/ethyl acetate = 6/4) to give 3- [ (4-fluoro-2-methoxy-5-nitrophenyl) carbonyl ] -1H indole 072f (150 mg, white solid), yield: 85 percent.
MS m/z(ESI)：315.2[M+1] + 。
The sixth step
Preparation of 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyaniline
3- [ (4-fluoro-2-methoxy-5-nitrophenyl) carbonyl ] -1H indole 072f (120mg, 0.38mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (213mg, 3.82mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and then extracted with dichloromethane (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 072g (100 mg, white solid) of 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyaniline, yield: 83 percent.
MS m/z(ESI)：285.2[M+1] + 。
Seventh step
Preparation of 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
072g (50mg, 0.18mmol) of 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyaniline was dissolved in tetrahydrofuran (10 mL), and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (71mg, 0.21mmol) and triethylamine (53mg, 0.53mmol) were added. The reaction solution was stirred at 25 ℃ for 16 hours. After LCMS monitoring the reaction was complete, the reaction gave compound 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 072H (100 mg, light yellow liquid), yield: 54 percent.
MS m/z(ESI)：593.1[M+1] + 。
Eighth step
Preparation of 3- { 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction mixture in the seventh step. After addition of lithium hydroxide monohydrate (12mg, 0.28mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give a crude product which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester Cpd-015 (20 mg, white solid) yield: and 43 percent.
MS m/z(ESI)：480.1[M+1] + 。
HPLC:99.19％(214nm),99.18％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.53(s,1H),12.06(s,1H),11.99(s,1H),8.19-8.15(m,1H),7.66(d,J＝3.2Hz,1H),7.55(d,J＝8.4Hz,1H),7.50(dd,J＝6.4,2.0Hz,1H),7.36(s,1H),7.30(d,J＝12.4Hz,1H),7.27-7.20(m,2H),3.81(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-117.01.
Example 16
Preparation of 3- (5- ((1H-indol-7-yl) methoxy) -2-fluoro-4-methoxyphenyl-2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-016)
Figure BDA0003608368340000491
First step of
1-tosyl-1H-indole-7-carbaldehyde
To tetrahydrofuran (20 mL) were added sodium hydride (124mg, 5.17mmol) and indole-7-carbaldehyde 78a (500mg, 3.44mmol) under ice-bath conditions, and after stirring at room temperature for 20 minutes, p-toluenesulfonyl chloride (985mg, 5.17mmol) was added to the mixture. The mixture was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was quenched by addition of saturated aqueous ammonium chloride (20 mL), extracted with ethyl acetate (2X 50 mL), and the organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated by silica gel column chromatography, eluted with n-hexane/ethyl acetate (4:1 (v/v)), and separated and purified to give 1-tosyl-1H-indole-7-carbaldehyde 78b (700 mg, white solid) in a yield of 65%.
MS m/z(ESI)：300.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.50(s,1H),7.91-7.84(m,2H),7.68(dd,J＝7.6,1.1Hz,1H),7.46(dd,J＝16.0,8.1Hz,3H),7.32(d,J＝8.0Hz,2H),7.03(d,J＝3.6Hz,1H),2.29(s,3H).
Second step of
(1-tosyl-1H-indol-7-yl) methanol
1-tosyl-1H-indole-7-carbaldehyde 78b (700mg, 2.32mmol) was dissolved in a solution of MeOH (10 mL), and sodium borohydride (26mg, 6.96mmol) was added thereto, and the reaction solution was stirred at 25 ℃ for 2 hours. After completion of the reaction, the reaction mixture was concentrated to remove the solvent, and water (20 mL) was added to the remaining organic solvent and extracted with EtOAc (3X 20 mL). The combined organic layers were washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to give (1-tosyl-1H-indol-7-yl) methanol 78c (520 mg, yield: 76%).
MS m/z(ESI)：324.1(M+23)。
1 H NMR(400MHz,DMSO-d 6 )δ7.79(d,J＝3.8Hz,1H),7.56(dd,J＝12.8,8.3Hz,3H),7.48(d,J＝7.2Hz,1H),7.36(d,J＝8.0Hz,2H),7.27(t,J＝7.6Hz,1H),6.89(d,J＝3.8Hz,1H),5.25(t,J＝28.4Hz,1H),4.79(d,J＝46.8Hz,2H),2.32(s,3H).
The third step
7- (bromomethyl) -1-tosyl-1H-indole
(1-tosyl-1H-indol-7-yl) methanol 78c (520mg, 1.76mmol) was dissolved in methylene chloride (6 mL), and tetrabromomethane (23mg, 0.07mmol) and triphenylphosphine (18mg, 0.07mmol) were added thereto. The reaction solution was stirred at room temperature for 4 hours. After completion of the reaction, the mixture was concentrated to give a crude residue. To the residue was added water and extracted with ethyl acetate (60 mL. Times.3). The organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated to give the crude product. Purification by column on silica gel (petroleum ether/ethyl acetate = 4/1) gave 7- (bromomethyl) -1-tosyl-1H-indole 78d (430 mg, off-white solid) in 66% yield.
1 H NMR(400MHz,CDCl 3 )δ7.74(d,J＝3.8Hz,1H),7.59(d,J＝8.4Hz,2H),7.47(dd,J＝7.8,1.2Hz,1H),7.43-7.37(m,1H),7.23(dd,J＝14.2,7.8Hz,3H),6.71(d,J＝3.8Hz,1H),5.15(s,2H),2.35(s,3H).
The fourth step
7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1-tosyl-1H-indole
7- (bromomethyl) -1-tosyl-1H-indole 78d (430mg, 1.16mmol) was dissolved in 5mL of an acetonitrile solution, and 4-fluoro-2-methoxy-5-nitrophenol (220mg, 1.18mmol) and potassium carbonate (195mg, 1.41mmol) were added thereto. The reaction solution was stirred at 70 ℃ for 2 hours. Water was added to the reaction solution and the mixture was extracted with DCM (3X 60 mL). The organic layers were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was isolated and purified by silica gel column (petroleum ether/ethyl acetate = 3/2) to give 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1- (4-methyl-1-sulfonylphenyl) indole 78e (500 mg, yellow solid) in 47% yield.
1 H NMR(400MHz,CDCl 3 )δ7.74(d,J＝3.8Hz,1H),7.62-7.44(m,5H),7.40(d,J＝7.2Hz,1H),7.29(d,J＝7.8Hz,1H),7.19(d,J＝8.0Hz,2H),6.75(d,J＝3.8Hz,1H),5.52(s,2H),3.95(s,3H),2.33(s,3H).
The fifth step
2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) aniline
7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1- (4-methyl-1-sulfonylphenyl) indole 78e (500mg, 1mmol) was dissolved in MeOH (5 mL), and palladium/carbon (50 mg) was added thereto, the reaction system was placed under a hydrogen balloon atmosphere, and the air in the reaction system was replaced. After the reaction was complete, the mixture was filtered off palladium on carbon, the filter cake was washed with methanol (3 × 30 mL), and the organic phase was concentrated to give 2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) aniline 78f (300 mg, as a pale yellow solid) in 60% yield. Directly used for the next reaction.
MS m/z(ESI)：441.1[M+1] + 。
The sixth step
4- (3- (2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
To a solution of 2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) aniline 78f (300mg, 1mmol) in THF (5 mL) were added 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (522mg, 1.53mmol) and triethylamine (1.05g, 10.35mmol), and the reaction was stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, water was added to the residue and the mixture was extracted with ethyl acetate (3 × 60 mL). The organic layers were combined, washed with saturated sodium chloride solution, the organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, which was further purified with a silica gel column (petroleum ether/ethyl acetate = 4/1) to give 78g (250 mg, white solid) of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate in 37% yield.
MS m/z(ESI)：682.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.97(s,1H),8.97(s,1H),8.86(s,1H),8.86(s,1H),7.97-7.82(m,2H),7.98-7.83(m,2H),7.74-7.57(m,4H),7.73-7.58(m,4H),7.52(d,J＝7.2Hz,1H),7.52(d,J＝7.2Hz,1H),7.37-7.20(m,3H),7.39-7.19(m,3H),6.98(dd,J＝26.4,8.2Hz,2H),6.98(dd,J＝26.4,8.2Hz,2H),5.35(s,2H),5.35(s,2H),3.89(s,4H),3.86(d,J＝27.2Hz,9H),3.83(s,4H),2.26(s,2H),2.26(s,3H).
Seventh step
3- (5- ((1H-indol-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate (250mg, 0.36mmol) was dissolved in a solvent (tetrahydrofuran/methanol = 2/1), and cesium carbonate (586 mg,1.8 mmol) was added thereto, and the reaction solution was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, water (10 mL) was added to the residue, and the mixture was extracted with ethyl acetate (3X 30 mL). The organic layers were combined, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was purified by HPLC (mobile phase: acetonitrile/water) preparation to give 3- (5- ((1H-indol-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-016 (10 mg, yellow solid), yield: 7 percent.
MS m/z(ESI)：482.1[M+1] + 。
HPLC:98.84％(214nm),98.18％(254nm)。
1 H NMR(400MHz,DMSO-d 6 )δ14.58(s,1H),11.95(s,1H),11.25(s,1H),7.57(d,J＝7.9Hz,1H),7.36-7.33(m,2H),7.18-7.12(m,2H),7.00(t,J＝7.5Hz,1H),6.49-6.48(m,2H),5.23(s,2H),3.80(s,3H)。
Example 17
Preparation of (3- (2-fluoro-4-methoxy-5- (quinoline-8-methoxy) phenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-017)
Figure BDA0003608368340000511
First step of
Preparation of 8-hydroxymethylquinoline
8-Formylquinoline (1g, 6.36mmol) was dissolved in MeOH Alcohol (10 mL), then NaBH was added 4 (720mg, 3mmol). After reaction at room temperature for 30min, the reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (3X 10 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 8-hydroxymethylquinoline 080b (0.8 g, grey solid), yield: 71 percent.
MS m/z(ESI)：160.2[M+1] + .
Second step of
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline
4-fluoro-2-methoxy-5-nitrophenol (500mg, 2.67mmol), 8-hydroxymethylquinoline (425.3mg, 2.67mmol) and PPh3 (911.1mg, 3.47mmol) were dissolved in dry THF (10 mL) solution, followed by the addition of DIAD (702.4mg, 3.47mmol). Reacting at 45 ℃ for 2h. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE: etOAc = 25%) to give 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline 080c (380 mg, yellow solid) in yield: 39 percent of
MS m/z(ESI)：329.1[M+1] + .
The third step
Preparation of 2-fluoro-4-methoxy-5- (quinoline-8-methoxy) aniline
Compound 080c (370mg, 1.13mmol) dissolved in MeOH (10 mL) and saturated NH 4 To the combined solution of Cl (2 mL) was then added iron powder (629mg, 11.27mmol). Reacting at room temperature for 2h, filtering, and concentrating the filtrate under reduced pressure. The resulting residue was dissolved in ethyl acetate (20 mL) and saturated NaHCO was used 3 And NaCl solution, and drying the mixture by anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 2-fluoro-4-methoxy-5- (quinoline-8-methoxy) aniline 080d (330 mg, brown solid) in yield: 88 percent. Yield: 44 percent.
MS m/z(ESI)：299.2[M+1] + .
The fourth step
Preparation of 4- (3- (2-fluoro-4-methoxy-5- (quinolin-8-ylmethoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
Compound 080d (320mg, 1.07mmol) and triethylamine (326mg, 3.22mmol) were dissolved in THF (10 mL) and 4- ((phenoxycarbonyl) amino) thiophenedimethyl-2,3-dicarboxylic acid dimethyl ester (432mg, 1.2872mmol) was added. After reacting at room temperature for 16h, the reaction solution was concentrated under reduced pressure to give 4- (3- (2-fluoro-4-methoxy-5- (quinolin-8-ylmethoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 080e (450 mg, yellow oily liquid), yield: 62 percent.
MS m/z(ESI)：540.2[M+1] + .
The fifth step
Preparation of (3- (2-fluoro-4-methoxy-5- (quinoline-8-methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 080e dissolved (450mg, 0.83mmol) in THF/MeOH/H 2 To a mixed solution of O = 5. The reaction was carried out at room temperature for 2h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by preparative HPLC (ACN: H20 (0.2% FA) = 50%) to give (3- (2-fluoro-4-methoxy-5- (quinoline-8-methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d) ]Pyrimidine-5-carboxylic acid Cpd-017 (60 mg, light yellow solid), yield: 14.6 percent.
MS m/z(ESI)：494.1[M+1] + .
1 H NMR(400MHz,DMSO-d 6 )δ14.59(s,1H),11.98(s,1H),8.94-8.92(m,1H),8.43(dd,J＝8.2,1.6Hz,1H),8.00(d,J＝8.2Hz,1H),7.95(d,J＝6.2Hz,1H),7.70-7.65(m,1H),7.60(dd,J＝8.2,4.2Hz,1H),7.41(d,J＝7.2Hz,1H),7.38(s,1H),7.16(d,J＝11.6Hz,1H),5.64(s,2H),3.85(s,3H).
19 F NMR(376MHz,DMSO-d 6 )δ-128.91(s,1H).
Example 18
Preparation of 3- (2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-018)
Figure BDA0003608368340000531
First step of
Preparation of 5- (bromomethyl) quinoxaline
5-methylquinoxaline 081a (2.20g, 0.02mol) is dissolved in carbon tetrachloride (30 mL), and N-bromosuccinimide (2.72g, 0.02mol) and a catalytic amount of benzoyl peroxide are added to react at 80 ℃ for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was washed with water (100 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/80% petroleum ether) to give 5- (bromomethyl) quinoxaline 081b (2.80 g, yellow solid), yield: 82 percent.
MS m/z(ESI)：223.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.07(d,J＝1.8Hz,1H),9.03(d,J＝1.8Hz,1H),8.11(dd,J＝8.4,1.4Hz,1H),8.06(dd,J＝7.2,1.2Hz,1H),7.87(dd,J＝8.4,7.2Hz,1H),5.29(s,2H).
Second step of
Preparation of 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline
5- (bromomethyl) quinoxaline 081b (142mg, 0.64mmol) was dissolved in acetonitrile (10 mL), and potassium carbonate (106mg, 0.76mmol) and 4-fluoro-2-methoxy-5-nitrophenol (119mg, 0.64mmol) were added and reacted at 70 ℃ for 3 hours. After the reaction was completed, water (10 mL) was added for quenching, extraction was performed with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to obtain 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 081c (200 mg, white solid), yield: 95 percent.
MS m/z(ESI)：330.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.90(dd,J＝5.4,1.8Hz,2H),8.11(d,J＝8.4Hz,1H),7.99(d,J＝7.2Hz,1H),7.86(d,J＝7.2Hz,1H),7.83-7.79(m,1H),6.74(d,J＝12.4Hz,1H),5.86(s,2H),3.96(s,3H).
The third step
Preparation of 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline
5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 081c (257mg, 0.78mmol) is dissolved in ethanol (3 mL) and water (3 mL), iron powder (131mg, 2.34mmol) and ammonium chloride (125mg, 2.34mmol) are added, and the reaction solution is reacted at 80 ℃ for 1 hour. After the completion of the reaction, extraction was performed with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 4/1) to obtain 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline 081d (143 mg, white solid), yield: 61 percent.
MS m/z(ESI)：300.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.01(dd,J＝11.0,1.8Hz,2H),8.09(dd,J＝8.2,1.4Hz,1H),7.98-7.89(m,2H),6.81(d,J＝12.4Hz,1H),6.62(d,J＝8.8Hz,1H),5.63(s,2H),4.65(s,2H),3.68(s,3H).
The fourth step
Preparation of 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline 081d (113mg, 0.38mmol) is dissolved in tetrahydrofuran (5 mL), triethylamine (57mg, 0.57mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (152mg, 0.45mmol) are added, the reaction is stirred at 50 ℃ for 4 hours, after completion of the reaction, the reaction liquid is concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 081e (184 mg, white solid) with a yield of 90%.
MS m/z(ESI)：541.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.02(dd,J＝7.4,1.8Hz,2H),8.97(s,1H),8.83(s,1H),8.11(dd,J＝8.2,1.2Hz,1H),8.02(d,J＝6.0Hz,1H),7.94-7.90(m,2H),7.87(d,J＝8.0Hz,1H),7.04(d,J＝12.4Hz,1H),5.68(s,2H),3.88(s,3H),3.83(s,3H),3.76(s,3H).
The fifth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 081e (134mg, 0.25mmol) was dissolved in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL), and lithium hydroxide (31mg, 0.74mmol) was added to react at 25 ℃ for 2 hours. After the reaction was completed, dilute hydrochloric acid was added to adjust to pH =4-5 and extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-018 (36 mg, white solid), yield: 29 percent.
MS m/z(ESI)：495.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.58(s,1H),11.98(s,1H),9.01(d,J＝1.8Hz,1H),8.98(d,J＝1.8Hz,1H),8.12(dd,J＝8.4,1.2Hz,1H),8.04(d,J＝6.0Hz,1H),7.95-7.91(m,1H),7.41-7.38(m,2H),7.16(d,J＝11.6Hz,1H),5.64(s,2H),3.84(s,3H).
Example 19
Preparation of 3- (2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-019)
Figure BDA0003608368340000541
First step of
Preparation of 5-methyl quinazoline
To a solution of 5-bromoquinazoline 083a (1.00g, 4.80mmol) in N, N-dimethylformamide (20 mL) were added methylboronic acid (340mg, 5.70mmol), 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium (350mg, 0.40mmol) and potassium carbonate (710mg, 7.20mmol). The reaction mixture was stirred in a microwave oven at 80 ℃ for 1 hour. After the reaction was completed, water quenching was added, extraction was performed with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to obtain 5-methyl quinazoline 083b (299 mg, white solid) in yield: 44 percent.
MS m/z(ESI)：145.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.74(s,1H),9.32(s,1H),7.93-7.88(m,1H),7.86-7.84(m,1H),7.57(d,J＝6.8Hz,1H),2.78(s,3H).
Second step of
Preparation of 5- (bromomethyl) quinazoline
To a solution of 5-methyl quinazoline 083b (299mg, 2.07mmol) in carbon tetrachloride (10 mL) were added N-bromosuccinimide (443mg, 2.49mmol) and 2,2' -azobis (2-methylpropanenitrile) (34mg, 0.21mmol), and the reaction mixture was stirred at 85 ℃ under nitrogen for 3 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain 5- (bromomethyl) quinazoline 083c (290 mg, white solid) in yield: and 63 percent.
MS m/z(ESI)：223.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.93(s,1H),9.38(s,1H),8.04-7.98(m,2H),7.90(d,J＝6.4Hz,1H),5.37(s,2H).
The third step
Preparation of 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinazoline
To a solution of 4-fluoro-2-methoxy-5-nitrophenol (292mg, 1.56mmol) in acetonitrile was added potassium carbonate (270mg, 1.95mmol) and 5- (bromomethyl) quinazoline 083c (290mg, 1.30mmol), and the reaction mixture was stirred at 70 ℃ for 2 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 20/1) to give 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinazoline 083d (317 mg, yellow solid) in yield: 74 percent.
MS m/z(ESI)：330.0[M+1] + 。
The fourth step
Preparation of 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) aniline
To a solution of 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinazoline 083d (287 mg, 0.87mmol) in ethanol (5 mL) and water (5 mL) were added iron powder (146mg, 2.61mmol) and ammonium chloride (140mg, 2.61mmol), and the reaction mixture was stirred at 80 ℃ for 1 hour. After the reaction was completed, the reaction was filtered while it was hot, the filtrate was extracted with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated and dried to obtain 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) aniline 083e (217 mg, yellow solid), yield: 83 percent.
MS m/z(ESI)：300.1[M+1] + 。
The fifth step
Preparation of dimethyl 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
To a solution of 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) aniline 083e (167mg, 0.56mmol) in tetrahydrofuran (10 mL) was added triethylamine (85mg, 0.84mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (225mg, 0.67mmol), the reaction mixture was stirred at 25 ℃ for 8 hours, after completion of the reaction, water was added to quench, dichloromethane (3 × 20 mL) was used for extraction, the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester f (200 mg, white solid) with a yield of 66%.
MS m/z(ESI)：541.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.85(s,1H),9.35(s,1H),8.98(s,1H),8.83(s,1H),8.03-8.01(m,2H),7.93(s,1H),7.89-7.87(m,2H),7.05(d,J＝12.6Hz,1H),5.62(s,2H),3.89(s,3H),3.83(s,3H),3.72(s,3H).
The sixth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 083f (170mg, 0.31mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (23mg, 0.94mmol) and reacted at 25 ℃ for 2 hours. After the reaction was completed, 2N diluted hydrochloric acid was added dropwise to adjust pH =4-5, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-019 (56 mg, white solid) with yield: 36 percent.
MS m/z(ESI)：495.0[M+1] + 。
HPLC:99.35％(214nm),98.12％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ9.83(s,1H),9.35(s,1H),8.03-8.01(m,2H),7.88-7.85(m,1H),7.37(d,J＝7.4Hz,1H),7.15(d,J＝11.6Hz,1H),7.11(s,1H),5.62(s,2H),3.83(s,3H).
Example 20
Preparation of 3- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-020)
Figure BDA0003608368340000561
First step of
Preparation of 2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione
2-fluoro-4-methoxyaniline 085a (5.00g, 35.40mmol) and 2-benzofuran-1,3-dione (16.00g, 106.20mmol) were dissolved in DMF. The mixture was reacted at 120 ℃ for 1 hour in a microwave oven, then quenched by addition of water (50 mL), extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the residue obtained was separated and purified with a silica gel column (petroleum ether/ethyl acetate = 2/1) to give 2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione 085b (6.00 g, pink solid) in yield: and 55 percent.
MS m/z(ESI)：272.0[M+1] + 。
1 H NMR(400MHz,d 6- DMSO)δ8.01-7.98(m,2H),7.96-7.93(m,2H),7.47(t,J＝8.8Hz,1H),7.09(dd,J＝12.0,2.8Hz,1H),6.97-6.94(m,1H),3.84(s,3H).
Second step of
Preparation of 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
After 2- (2-fluoro-4-methoxyphenyl) isoindole-1,3-dione 085b (3.00g, 11.00mmol) was dissolved in dichloromethane (3 mL) and slowly added dropwise to chlorosulfonic acid (30 mL) and stirred at 25 ℃ for 4 hours, the reaction was quenched by slowly pouring into ice water and extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated to give 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 085c (3.10 g, pink solid) with a yield of 60%.
MS m/z(ESI)：392.0(M+23)。
1 H NMR(400MHz,d 6- DMSO)δ8.00-7.97(m,2H),7.94-7.91(m,2H),7.80(d,J＝9.2Hz,1H),7.11(d,J＝12.4Hz,1H),3.84(s,3H).
The third step
Preparation of 2- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] isoindole-1,3-dione
3-methyl-1H-indole (532mg, 4.06mmol) was dissolved in tetrahydrofuran (10 mL). Then sodium hydride (195mg, 8.11mmol) was added at 0 ℃. The mixture was stirred at 0 ℃ for 0.5 h, then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 085c (300mg, 0.81mmol) was dissolved in tetrahydrofuran (5 mL) and the solution was added dropwise with continued stirring at 25 ℃. After 1.5 hours, the reaction was quenched by slowly pouring it into ice water and adjusting its pH to 6-7 with 1M dilute hydrochloric acid, followed by extraction with dichloromethane (2X 30 mL), combining the organic phases and drying over anhydrous sodium sulfate, and purifying the concentrated residue by column chromatography (petroleum ether/ethyl acetate: 3/7) to give 2- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] isoindole-1,3-dione 085d (150 mg, white solid) in 35% yield.
MS m/z(ESI)：465.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.17(d,J＝8.0Hz,1H),7.99-7.98(m,2H),7.84-7.82(m,3H),7.49(d,J＝7.6Hz,1H),7.33-7.29(m,3H),6.75(d,J＝11.2Hz,1H),3.70(s,3H),2.28(s,3H)。
The fourth step
Preparation of 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) aniline
2- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] isoindole-1,3-dione 085d (140mg, 0.30mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The mixture was stirred at 90 ℃ for 1 hour, then the solvent was removed by spin-drying, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 2/3) to give preparation 085e (80 mg, orange solid) of 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) aniline, yield: 63%.
MS m/z(ESI)：335.0[M+1] + 。
The fifth step
Preparation of 4- ({ [ [ [ [ [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
The preparation of 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) aniline 085e (30mg, 0.09mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (72mg, 0.21mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (36mg, 0.36mmol) was added, the mixed solution was stirred at 25 ℃ for 16 hours, the reaction solution was not subjected to any post-treatment to obtain a stock solution of 4- ({ [ [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 085f (bright yellow solution), which was directly used in the next reaction.
MS m/z(ESI)：576.0[M+1] + 。
The sixth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution of the seventh step were added methanol (3 mL) and water (1 mL), followed by lithium hydroxide (10mg, 0.43mmol). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction is finished, the reaction product is directly concentrated, is primarily purified by a reverse column (water: acetonitrile = 3:2) and is then purified by a preparation (acetonitrile/0.1% formic acid water solution) to obtain the Cpd-020 (2.09 mg, white solid) prepared from the 3- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-formic acid, wherein the yield is 4%.
MS m/z(ESI)：530.0[M+1] + 。
HPLC:98.01％(214nm),97.90％(254nm)。
1 H NMR(400MHz,d 6- DMSO)δ11.89(s,1H),8.42(d,J＝8.0Hz,1H),7.64(dd,J＝6.0,2.8Hz,1H),7.58-7.56(m,1H),7.50(d,J＝1.2Hz,1H),7.34(d,J＝11.6Hz,1H),7.27-7.22(m,3H),3.77(s,3H),2.26(s,3H).
Example 21
Preparation of 3- [5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-021)
Figure BDA0003608368340000571
First step of
Preparation of 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
2H-1,4-benzoxazine (79mg, 0.6 mmol) and triethylamine (164mg, 1.60mmol) were dissolved in acetonitrile (5 mL). Then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 086a (200mg, 0.5mmol) was added. The reaction mixture was reacted at 100 ℃ for 3 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 086b (210 mg, white solid) in 75% yield.
MS m/z(ESI)：468.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.06(d,J＝8.0Hz,1H),7.98-7.92(m,2H),7.81(dd,J＝5.6,3.2Hz,2H),7.57(d,J＝8.4Hz,1H),7.03-6.97(m,1H),6.93-6.77(m,3H),4.10(q,J＝7.2Hz,2H),3.94-3.89(m,2H),3.63(s,3H).
Second step of
Preparation of 5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyaniline
5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 086b (210mg, 0.448mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction mixture was reacted at 90 ℃ for 3 hours. Then extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyaniline 086c (110 mg, white solid) in 65% yield.
MS m/z(ESI)：339[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.57-7.47(m,2H),7.01-6.96(m,1H),6.88-6.83(m,2H),6.64(d,J＝12.0Hz,1H),4.04-4.00(m,2H),3.92-3.88(m,2H),3.53(s,3H).
The third step
Preparation of dimethyl 4- ({ [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
(5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyaniline 086c (80mg, 0.24mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (237 mg, 0.71mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (71mg, 0.71mmol) was added, and the reaction solution was reacted at 25 ℃ for 1 hour to obtain 4- ({ [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 086d without any post-treatment.
MS m/z(ESI)：580.1[M+1] + 。
The fourth step
Preparation of 3- [5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution in the third step, methanol (3 mL) and water (1 mL) were added, followed by lithium hydroxide (6 mg, 0.14mmol). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction is finished, the reaction product is directly concentrated, is primarily purified by a reverse column (water: acetonitrile = 60), and is then purified by a preparation (acetonitrile/0.1% formic acid water solution) to obtain 3- [5- (2,3-dihydro-1,4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-021 (25 mg, white solid) with the yield of 52%.
MS m/z(ESI)：534.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.37(s,1H),8.09(d,J＝8.4Hz,1H),7.44-7.31(m,2H),7.04-6.97(m,1H),6.90-6.82(m,2H),6.63(s,1H),4.13-3.96(m,2H),3.94-3.81(m,2H),3.69(s,3H).
Example 22
Preparation of 3- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoline-1-sulfonyl) phenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-022)
Figure BDA0003608368340000591
First step of
Preparation of 3,4-dihydro-2H-quinoxaline-1-carboxylic acid tert-butyl ester
1,2,3,4-tetrahydroquinoxaline 087a (500mg, 3.72mmol) was dissolved in dichloromethane (20 mL) and triethylamine (1.13g, 11.18mmol), N, N-dimethylpyridin-4-amine (45mg, 0.37mmol) and B-anhydride (813mg, 3.72mmol) were added. The reaction solution was stirred at 25 ℃ for 3 hours. After completion of the reaction, the solvent was removed by concentration under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 80/20) to give 3,4-dihydro-2H-quinoxaline-1-carboxylic acid tert-butyl ester 087b (300 mg, light yellow oil), yield: 33 percent.
MS m/z(ESI)：257.1(M+Na)。
Second step of
Preparation of 4-methyl-2,3-dihydroquinoxaline-1-carboxylic acid tert-butyl ester
3,4-dihydro-2H-quinoxaline-1-carboxylic acid tert-butyl ester 087b (700mg, 2.99mmol) is dissolved in acetonitrile (3 mL), iodomethane (826mg, 5.97mmol) and potassium carbonate (825mg, 5.97mmol) are then added, stirring is carried out at 80 ℃ for 16 hours, after the reaction is completed, water (10 mL) is added to the reaction mixture, extraction is carried out three times with dichloromethane, the organic phase is washed with saturated brine (20 mL), drying is carried out with anhydrous sodium sulfate, and then concentration is carried out to obtain a crude product, which is purified by a silica gel column (petroleum ether/ethyl acetate = 85/15) to obtain 4-methyl-2,3-dihydroquinoxaline-1-carboxylic acid tert-butyl ester 087c (550 mg, brown oil) in yield: 38 percent.
MS m/z(ESI)：271.1(M+Na)。
The third step
Preparation of 1-methyl-3,4-dihydro-2H-quinoxaline
4-methyl-2,3-dihydroquinoxaline-1-carboxylic acid tert-butyl ester 087c (300mg, 1.21mmol) was dissolved in dichloromethane (5 mL) followed by the addition of trifluoroacetic acid (1 mL). The reaction mixture was stirred at 25 ℃ for 2 hours. LCMS monitoring completion of the reaction, concentration under reduced pressure gave 1-methyl-3,4-dihydro-2H-quinoxaline preparation 087d (200 mg, brown oil), yield: 89 percent.
MS m/z(ESI)：149.2[M+1] + 。
The fourth step
Preparation of 2- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) phenyl ] isoindole-1,3-dione
Preparation of 1-methyl-3,4-dihydro-2H-quinoxaline 087d (250mg, 0.10mmol) and 5- (1,3-dioxoisopropanol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (686mg, 1.86mmol) were dissolved in 1,2-dichloroethane (10 mL), followed by the addition of DIPEA (654mg, 5.06mmol) and pyridine (267mg, 3.37mmol). The reaction solution was stirred at 100 ℃ for 1 hour. After completion of the reaction, water (10 mL) was added to the reaction solution, extraction was performed with dichloromethane (3 × 20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to obtain a crude product, which was subjected to silica gel column (petroleum ether/ethyl acetate = 4/6) to obtain 2- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) phenyl ] isoindole-1,3-dione 087e (200 mg, white solid) in yield: 12 percent.
MS m/z(ESI)：482.2[M+1] + 。
The fifth step
Preparation of 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) aniline
2- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoline-1-sulfonyl) phenyl ] isoindole-1,3-dione 087e (100mg, 0.22mmol) was dissolved in ethanol (5 mL), followed by addition of hydrazine hydrate (1 mL). The reaction solution was stirred at 80 ℃ for 2 hours. After completion of the reaction, water (20 mL) was added to the reaction solution, extraction was performed with dichloromethane (3 × 20 mL), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to obtain a crude product, and preparation of 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) aniline through silica gel column (petroleum ether/ethyl acetate = 3/7) 30g (50 mg, white solid) was obtained in yield: and 63 percent.
MS m/z(ESI)：374.2[M+1] + 。
The sixth step
Preparation of 4- ({ [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) phenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
30g (50mg, 0.14mmol) of 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) aniline was dissolved in tetrahydrofuran (20 mL), 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (57mg, 0.17mmol) and triethylamine (43mg, 0.43mmol) were added, and the reaction solution was stirred at 25 ℃ for 16 hours. After completion of the reaction, LCMS monitored, this step was used directly in the next step without work-up.
MS m/z(ESI)：593.1[M+1] + 。
The ninth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) phenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution in the sixth step were added 2mL and 5mL of methanol, and lithium hydroxide (24mg, 0.56mmol) was added and the reaction solution was stirred at 25 ℃ for 3 hours. After the reaction was completed by LCMS monitoring, the reaction was concentrated to remove the organic solvent and the residue was subjected to preparative (acetonitrile/water/0.1% formic acid) to give 3- [ 2-fluoro-4-methoxy-5- (4-methyl-2,3-dihydroquinoxaline-1-sulfonyl) phenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-022 (8 mg, orange solid), yield: 17 percent.
MS m/z(ESI)：547.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,CD 3 OD)δ8.05(d,J＝8.0Hz,1H),7.34-7.29(m,2H),7.10(d,J＝11.6Hz,1H),7.05-6.99(m,1H),6.69(d,J＝7.6Hz,1H),6.59(t,J＝7.6Hz,1H),3.78(dd,J＝6.0,4.8Hz,2H),3.48(s,3H),3.03(t,J＝5.6Hz,2H),2.82(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-77.43,-111.21.
Example 23
Preparation of 3- [5- (4-Chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-023)
Figure BDA0003608368340000601
First step of
Preparation of 2- [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione
4-chloro-1H-indole 088a (135mg, 0.89mmol) was dissolved in tetrahydrofuran (5 mL). Sodium hydride (58mg, 2.43mmol) was then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 0.5 hour. Then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (300mg, 0.81mmol) was dissolved in tetrahydrofuran (2 mL), and the solution was added dropwise thereto, and the reaction solution was reacted at 25 ℃ for 3 hours. Then, the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, methylene chloride (2X 10 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 2- [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 088b (112 mg, white solid) in 26% yield.
MS m/z(ESI)：503.0[M+1] + 。
Second step of
Preparation of 2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione
2- [5- (4-Chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 088b (89mg, 0.18mmol) was dissolved in acetic acid (2 mL). Then sodium acetate (43mg, 0.53mmol) was added. The reaction mixture was reacted at 90 ℃ for 1.5 hours. The pH was then adjusted to about 7 with saturated sodium bicarbonate solution, extracted with ethyl acetate (2X 5 mL), and the organic phases were combined and dried over anhydrous sodium sulfate to give 2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione 088c (70 mg, white solid) in 79% yield.
MS m/z(ESI)：485.0[M+1] + 。
The third step
Preparation of 5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline
2- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione 088c (80mg, 0.16mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction mixture was reacted at 90 ℃ for 4 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 088d (26 mg, white solid) in 40% yield.
MS m/z(ESI)：355.0[M+1] + 。
The fourth step
Preparation of dimethyl 4- ({ [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 088d (80mg, 0.23mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (90mg, 0.27mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (68mg, 0.67mmol) was added, the reaction solution was reacted at 25 ℃ for 1 hour, and the reaction solution was not subjected to any post-treatment to give a stock solution of 4- ({ [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 088e.
MS m/z(ESI)：596.0[M+1] + 。
The fifth step
Preparation of 3- [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution in the fourth step were added methanol (3 mL) and water (1 mL), followed by lithium hydroxide (5mg, 0.14mmol). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated, primarily purified by reverse column (water: acetonitrile = 60), and then purified by preparative method (acetonitrile/0.1% formic acid aqueous solution) to obtain 3- [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-023 (2.52 mg, white solid) with a yield of 9%.
MS m/z(ESI)：550.0[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ12.01(s,1H),8.53(d,J＝8.0Hz,1H),7.88(d,J＝3.6Hz,1H),7.62(d,J＝8.4Hz,1H),7.38(dd,J＝11.6,9.6Hz,3H),7.26(t,J＝8.0Hz,1H),6.84(d,J＝3.6Hz,1H),3.82(s,3H).
Example 24
Preparation of 3- {5- [ (5-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-024)
Figure BDA0003608368340000621
First step of
Preparation of 8-bromo-5-chloroisoquinoline
5-chloroisoquinoline 089a (1g, 6.1mmol) was dissolved in sulfuric acid (15 mL), N-bromosuccinimide (1.19g, 6.7mmol) was added under ice-water bath, and the reaction was stirred at 25 ℃ for 16 hours. The reaction was quenched by addition of water (100 mL), adjusted to pH 8 by addition of aqueous ammonia, and extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 4/1) to give the title product 8-bromo-5-chloroisoquinoline 089b (1.5 g, white solid) in 91% yield.
MS m/z(ESI)：242.1[M+1] + .
Second step of
Preparation of methyl 5-chloroisoquinoline-8-carboxylate
In an autoclave, 8-bromo-5-chloroisoquinoline 089b (1.5g, 6.2mmol) was dissolved in methanol (30 mL) and 1,1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride dichloromethane complex (0.51g, 0.6 mmol) and sodium acetate (0.76g, 9.3mmol) were added. The reaction mixture was stirred at 60 ℃ for 8 hours under a carbon monoxide atmosphere and 10 atmospheres of pressure. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate = 4/1) to obtain the title product methyl 5-chloroisoquinoline-8-carboxylate 089c (1.1 g, white solid) in a yield of 75%.
MS m/z(ESI)：222.2[M+1] + .
The third step
Preparation of (5-chloroisoquinolin-8-yl) methanol
Methyl 5-chloroisoquinoline-8-carboxylate 089c (500mg, 2.26mmol) was dissolved in tetrahydrofuran (10 mL) methanol (2 mL) and sodium borohydride (256mg, 6.77mmol) was added under an ice-water bath. The reaction solution was stirred at 25 ℃ for 16 hours. The reaction was quenched with water (100 mL) and extracted with ethyl acetate (3X 50 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the title product (5-chloroisoquinolin-8-yl) methanol 089d (400 mg, yellow solid) was obtained in 73% yield.
MS m/z(ESI)：194.1[M+1] + .
The fourth step
Preparation of 5-chloro-8- (chloromethyl) isoquinoline
(5-Chloroisoquinolin-8-yl) methanol 089d (400mg, 2.07mmol) was dissolved in thionyl chloride (10 mL). The reaction solution was stirred at 25 ℃ for 2 hours. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to give the title product 5-chloro-8- (chloromethyl) isoquinoline 089e (350 mg, yellow solid) in a yield of 71%.
MS m/z(ESI)：212.1[M+1] + .
The fifth step
Preparation of 5-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
4-fluoro-2-methoxy-5-nitrophenol (106mg, 0.57mmol) was dissolved in N, N-dimethylformamide (3 mL), and 5-chloro-8- (chloromethyl) isoquinoline 089e (120mg, 0.57mmol), potassium carbonate (157mg, 1.13mmol) and potassium iodide (94mg, 0.57mmol) were added. The reaction solution was stirred at 80 ℃ for 2 hours. The reaction was quenched with water (50 mL) and extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate = 1/1) to give the title product 5-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 089f (90 mg, yellow solid) in 39% yield.
MS m/z(ESI)：363.0[M+1] + .
The sixth step
Preparation of 5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
(5-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 089f (90mg, 0.25mmol) was dissolved in ethanol (4 mL) and water (1 mL), iron powder (70mg, 1.24mmol) and ammonium chloride (67mg, 1.24mmol) were added, the reaction was stirred at 90 ℃ for 6 hours, filtered, and the residue obtained by concentrating the filtrate was purified with a silica gel column (dichloromethane/methanol = 10/1) to give the title product 5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 089g (30 mg, yellow oil) in 34% yield.
MS m/z(ESI)：333.0[M+1] + .
Seventh step
Preparation of 4- [ ({ {5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
5- [ (5-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 089g (20mg, 0.06mmol) was dissolved in tetrahydrofuran (3 mL), and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (20mg, 0.06mmol) and triethylamine (19mg, 0.18mmol) were added in this order, and the reaction mixture was stirred at 25 ℃ for 16 hours, and the reaction mixture was concentrated to give the title product 4- [ ({ {5- [ (5-Chloroisoisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 089h (30 mg, yellow oil) in 60% yield.
MS m/z(ESI)：574.1[M+1] + .
Eighth step
Preparation of 3- {5- [ (5-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ {5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 089h (30mg, 0.05mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (111mg, 0.26mmol) was added, and the reaction was stirred at 25 ℃ for 2 hours. Water (0.5 mL) was then added and the reaction was stirred at 25 ℃ for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution and extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by preparative HPLC (acetonitrile/water (0.1% formic acid) to give the title product 3- {5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-024 (5.3 mg, white solid) in 19% yield.
MS m/z(ESI)：528.2[M+1] + .
HPLC:100％(214nm),98.17％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.52(s,1H),11.93(s,1H),9.61(s,1H),8.73(d,J＝6.0Hz,1H),8.07(d,J＝6.0Hz,1H),7.99(d,J＝7.6Hz,1H),7.79(d,J＝7.6Hz,1H),7.41(d,J＝7.2Hz,1H),7.31(s,1H),7.17(d,J＝11.6Hz,1H),5.62(s,2H),3.84(s,3H).
Example 25
Preparation of 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-025)
Figure BDA0003608368340000641
First step of
Preparation of [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine
After stirring a solution of 2-bromo-5-fluorobenzaldehyde 090a (5.00g, 0.02mol) and 2,2-dimethoxyethylamine (3.88g, 0.04mol) in ethanol (30 mL) at 80 ℃ for 8 hours, sodium cyanoborohydride (1.55g, 0.02mol) and a catalytic amount of acetic acid (1 mL) were added and stirred at 25 ℃ for 1 hour. After completion of the reaction, saturated aqueous ammonium chloride solution was added and quenched, extracted with ethyl acetate (3 × 50 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 090b (5.18 g, colorless oil) in yield: 72 percent.
MS m/z(ESI)：292.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.48(dd,J＝8.8,5.2Hz,1H),7.19(dd,J＝9.4,3.2Hz,1H),6.85(td,J＝8.4,3.2Hz,1H),4.50(t,J＝5.6Hz,1H),3.86(s,2H),3.39(s,6H),2.76(d,J＝5.6Hz,2H).
Second step of
Preparation of [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) [ (4-sulfonylphenyl) methyl ] amine
To a solution of [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 090b (5.18g, 0.02mol) and triethylamine (2.69g, 0.03mol) in dichloromethane (30 mL) was added p-toluenesulfonyl chloride (3.71g, 0.02mol) dropwise at 0 ℃ and reacted at 0 ℃ for 3 hours. After completion of the reaction, water (20 mL) was added to quench, extraction was performed with ethyl acetate (3 × 50 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 20/1) to give [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) [ (4-sulfonylphenyl) methyl ] amine 090c (7.80 g, white solid), yield: 98 percent.
MS m/z(ESI)：470.1(M+23)。
1 H NMR(400MHz,d 6 -DMSO)δ7.77(d,J＝8.0Hz,2H),7.63(dd,J＝8.8,5.4Hz,1H),7.46(d,J＝8.0Hz,2H),7.28(dd,J＝10.0,3.2Hz,1H),7.15-7.12(m,1H),4.37(s,2H),4.33(t,J＝5.2Hz,1H),3.28(d,J＝5.2Hz,2H),3.14(s,6H),2.42(s,3H).
The third step
Preparation of 8-bromo-5-fluoroisoquinoline
A solution of [ (2-bromo-5-fluorophenyl) methyl ] (2,2-dimethoxyethyl) [ (4-sulfonylphenyl) methyl ] amine 090c (5.00g, 0.01mol) (10 mL) in methylene chloride (10 mL) was added dropwise to a solution of aluminum trichloride (6.75g, 0.06mol) in methylene chloride (10 mL) at 0 ℃ and, after completion of the dropwise addition, reacted at 0 ℃ for 8 hours. After completion of the reaction, the reaction solution was slowly added dropwise to ice water to quench, and extracted with dichloromethane (3X 50 mL). The organic phase was washed with a saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 4/1) to give 8-bromo-5-fluoroisoquinoline 090d (1.13 g, white solid) in yield: 45 percent.
MS m/z(ESI)：226.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ9.49(s,1H),8.76(d,J＝5.8Hz,1H),8.02(dd,J＝8.4,4.8Hz,1H),7.97(dd,J＝5.8,0.6Hz,1H),7.62(dd,J＝9.8,8.4Hz,1H).
The fourth step
Preparation of methyl 5-fluoroisoquinoline-8-carboxylate
To a solution of 8-bromo-5-fluoroisoquinoline 090d (1.13g, 5.00mmol) and triethylamine (1.52g, 15.00mmol) in methanol (10 mL) was added [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) (0.37g, 0.50mmol) and reacted under 1atm carbon monoxide at 80 ℃ for 8 hours. After completion of the reaction, water was added) (10 mL) and the mixture was quenched and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give methyl 5-fluoroisoquinoline-8-carboxylate 090e (474 mg, yellow solid) in yield: 46 percent.
MS m/z(ESI)：206.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ10.16(s,1H),8.73(d,J＝5.8Hz,1H),8.33-8.29(m,1H),8.02(d,J＝5.8Hz,1H),7.76-7.71(m,1H),3.99(s,3H).
The fifth step
Preparation of (5-fluoroisoquinolin-8-yl) methanol
To a solution of methyl 5-fluoroisoquinoline-8-carboxylate 090e (400mg, 1.95mmol) in methanol (5 mL) was added sodium borohydride (221mg, 5.85mmol) at 0 ℃ and the mixture was reacted at 0 ℃ for 8 hours. After completion of the reaction, the reaction mixture was quenched dropwise with saturated aqueous ammonium chloride (5 mL) and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by reverse phase column chromatography to give (5-fluoroisoquinolin-8-yl) methanol 090f (90 mg, yellow solid) in yield: 26 percent.
MS m/z(ESI)：178.1[M+1] + 。
The sixth step
Preparation of 8- (bromomethyl) -5-fluoroisoquinoline
To a solution of (5-fluoroisoquinolin-8-yl) methanol 090f (90mg, 0.51mmol) in dichloromethane (3 mL) was added dropwise phosphine tribromide (138mg, 0.51mmol) at 0 ℃ and the reaction was carried out at 0 ℃ for 2 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added to adjust pH =6-7, and extraction was performed with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to give 090g (100 mg, white solid) of 8- (bromomethyl) -5-fluoroisoquinoline, yield: the content of the active carbon is 82%,
MS m/z(ESI)：240.0[M+1] + 。
seventh step
Preparation of 5-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
To a solution of 4-fluoro-2-methoxy-5-nitrophenol (62mg, 0.33mmol) and potassium carbonate (69mg, 0.50mmol) in acetonitrile (5 mL) was added 090g (80mg, 0.33mmol) of 8- (bromomethyl) -5-fluoroisoquinoline, and the mixture was reacted at 25 ℃ for 8 hours. After completion of the reaction, it was quenched by addition of water (5 mL) and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 5-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 090h (100 mg, white solid) in yield: 87 percent.
MS m/z(ESI)：347.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ9.63(s,1H),8.70(d,J＝5.8Hz,1H),8.00(dd,J＝10.2,6.6Hz,2H),7.83(dd,J＝8.0,5.4Hz,1H),7.67(dd,J＝10.2,8.0Hz,1H),7.31(d,J＝13.4Hz,1H),5.76(s,2H),3.88(s,3H).
Eighth step
Preparation of 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyaniline
To a solution of 5-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 090h (80mg, 0.23mmol) in ethanol (2 mL) and water (2 mL) were added iron powder (39mg, 0.69mmol) and ammonium chloride (37mg, 0.69mmol), and the mixture was reacted at 80 ℃ for 1 hour. After completion of the reaction, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to obtain 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyaniline 090i (56 mg, yellow solid), yield: 77 percent.
MS m/z(ESI)：317.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ9.62(s,1H),8.68(d,J＝5.6Hz,1H),7.97(d,J＝5.6Hz,1H),7.76-7.72(m,1H),7.65-7.60(m,1H),6.81(d,J＝12.4Hz,1H),6.64(d,J＝8.8Hz,1H),5.50(s,2H),4.66(s,2H),3.63(s,3H).
The ninth step
Preparation of 4- [ ({ { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
To a solution of 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyaniline 090i (46mg, 0.15mmol) and triethylamine (44mg, 0.44mmol) in tetrahydrofuran (3 mL) were added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (59mg, 0.17mmol), reacted at 25 ℃ for 3 hours, quenched after completion of the reaction, with water (5 mL), extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give dimethyl 4- [ ({ { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 090j (78 mg, yellow solid) in 96% yield.
MS m/z(ESI)：558.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ9.62(s,2H),8.83(s,1H),8.68(d,J＝5.8Hz,2H),7.97(d,J＝5.8Hz,2H),7.93(s,1H),7.87(d,J＝8.0Hz,1H),7.04(d,J＝12.6Hz,1H),5.56(s,2H),3.89(s,3H),3.83(s,3H),3.72(s,3H).
The tenth step
Preparation of 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of dimethyl 4- [ ({ { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 090j (78mg, 0.14mmol) in tetrahydrofuran (1 mL), methanol (1 mL) and water (1 mL) was added lithium hydroxide monohydrate (18mg, 0.42mmol), and the reaction was carried out at 25 ℃ for 3 hours. After completion of the reaction, 1M diluted hydrochloric acid was added dropwise to adjust pH =4-5, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give preparation Cpd-025 (11 mg, white solid) of 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid: 16 percent.
MS m/z(ESI)：512.0[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.54(s,1H),12.00(s,1H),9.60(s,1H),8.68(d,J＝5.8Hz,1H),7.98(d,J＝5.8Hz,1H),7.80(dd,J＝7.8,5.4Hz,1H),7.63(dd,J＝10.0,8.0Hz,1H),7.42-7.37(m,2H),7.17(d,J＝11.4Hz,1H),5.57(s,2H),3.83(s,3H).
Example 26
Preparation of 3- (2-fluoro-4-methoxy-5- ((quinoxalin-5-ylmethyl) amino) phenyl) -2,4 dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-026)
Figure BDA0003608368340000661
First step of
Preparation of 5- (bromomethyl) quinoxaline
5-Methylquinoxaline 091a (2.20g, 0.02mol) is dissolved in carbon tetrachloride (30 mL), and N-bromosuccinimide (2.72g, 0.02mol) and a catalytic amount of benzoyl peroxide are added to react at 80 ℃ for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was washed with water (100 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/50% petroleum ether) to give 5- (bromomethyl) quinoxaline 091b (2.80 g, yellow solid) in yield: 82 percent.
MS m/z(ESI)：223.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.07(d,J＝1.8Hz,1H),9.03(d,J＝1.8Hz,1H),8.11(dd,J＝8.4,1.4Hz,1H),8.06(dd,J＝7.2,1.2Hz,1H),7.87(dd,J＝8.4,7.2Hz,1H),5.29(s,2H).
Second step of
Preparation of 4-fluoro-2-methoxy-5-nitro-N- (quinoxalin-5-ylmethyl) aniline
5- (bromomethyl) quinoxaline 091b (1.70g, 7.60mmol) was dissolved in acetonitrile (20 mL), and potassium carbonate (1.26g, 9.10 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (1.41g, 7.60mmol) were added to react at 70 ℃ for 3 hours. After the reaction was completed, water (10 mL) was added to quench, a solid was precipitated, filtered, and the obtained filter cake was dried to obtain 4-fluoro-2-methoxy-5-nitro-N- (quinoxalin-5-ylmethyl) aniline 091c (1.75 g, brown solid), yield: 70 percent.
MS m/z(ESI)：329.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.04(dd,J＝4.6,1.8Hz,2H),8.01(dd,J＝8.0,2.0Hz,1H),7.84-7.78(m,2H),7.09(d,J＝13.2Hz,1H),7.03(d,J＝7.6Hz,1H),6.32(s,1H),5.02(d,J＝4.4Hz,2H),3.98(s,3H).
The third step
Preparation of 4-fluoro-6-methoxy-1-N- (quinoxalin-5-ylmethyl) benzene-1,3-diamine
4-fluoro-2-methoxy-5-nitro-N- (quinoxalin-5-ylmethyl) aniline 091c (1.70g, 5.20mmol) was dissolved in ethanol (5 mL) and water (5 mL), iron powder (0.87g, 15.60mmol) and ammonium chloride (0.83g, 15.60mmol) were added, and the reaction solution was reacted at 80 ℃ for 1 hour. After the reaction was completed, extraction was performed with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to obtain 4-fluoro-6-methoxy-1-N- (quinoxalin-5-ylmethyl) benzene-1,3-diamine 091d (728 mg, yellow solid), yield: 46 percent.
MS m/z(ESI)：299.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.00(s,2H),7.98(d,J＝8.4Hz,1H),7.84-7.78(m,1H),7.71(d,J＝6.8Hz,1H),6.65(d,J＝12.4Hz,1H),5.93(d,J＝8.8Hz,1H),5.35(t,J＝6.2Hz,1H),4.87(d,J＝6.2Hz,2H),4.30(s,2H),3.70(s,3H).
The fourth step
Preparation of dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoxalin-5-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
4-fluoro-6-methoxy-1-N- (quinoxalin-5-ylmethyl) benzene-1,3-diamine 091d (628mg, 2.11mmol) was dissolved in tetrahydrofuran (5 mL), triethylamine (639mg, 6.32mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (847mg, 2.53mmol) were added, the reaction was stirred at 50 ℃ for 5 hours, after completion of the reaction, the reaction solution was concentrated and purified by column chromatography (mobile phase: 60% ethyl acetate/40% petroleum ether) to give 4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoxalin-5-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate dimethyl 091e (1.02 g, yellow solid) in 90% yield.
MS m/z(ESI)：540.2[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.01-8.99(m,2H),8.65(s,2H),7.98(d,J＝6.8Hz,1H),7.82-7.75(m,3H),7.09(d,J＝8.0Hz,1H),6.85(d,J＝12.4Hz,1H),5.53(t,J＝6.0Hz,1H),4.92(d,J＝6.2Hz,2H),3.81(s,3H),3.80(s,3H),3.79(s,3H).
The fifth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoxalin-5-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 091e (200mg, 0.37mmol) was dissolved in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL), and lithium hydroxide (47mg, 1.11mmol) was added and reacted at 25 ℃ for 4 hours. After the reaction was completed, dilute hydrochloric acid was added to adjust pH =4-5 and extracted with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by reverse phase column chromatography (mobile phase: 30% acetonitrile/70% water) to give 3- [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-026 (44 mg, yellow solid) with a yield: 24 percent.
MS m/z(ESI)：494.1[M+1] + 。
HPLC:94.27％(214nm),88.60％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.11(s,1H),8.97(s,2H),7.99-7.96(m,1H),7.81-7.79(m,1H),6.91(d,J＝11.4Hz,1H),6.48(s,2H),5.50(s,1H),4.87(d,J＝5.8Hz,1H),3.85(s,3H).
Example 27
Preparation of 3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-027)
Figure BDA0003608368340000681
First step of
Preparation of 2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid
Sodium hydride (132mg, 2.43mmol) and 6-fluoroindole (151mg, 0.97mmol) were dissolved in tetrahydrofuran (10 mL) under ice-bath conditions, and after stirring at room temperature for 20 minutes, 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 092a (300mg, 0.81mmol) was added. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (5 mL) and extracted with ethyl acetate (2X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 4/1) to give 092b 2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid (158 mg, yellow solid) in a yield of 40%.
MS m/z(ESI)：487.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.80(s,1H),7.84(d,J＝7.2Hz,1H),7.73-7.65(m,3H),7.54(d,J＝5.0Hz,2H),7.41-7.45(m,1H),7.27(d,J＝12.1Hz,1H),7.13-7.10(m,1H),6.80-6.76(m,1H),3.69(s,3H)。
Second step of
Preparation of 2- (2-fluoro-5- (6-fluoro-1H-indol-1-yl) sulfanilamide) -4-methoxybenzene) isoindole-1,3-dione
092b (158mg, 0.32mmol) of 2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid and sodium acetate (79mg, 0.97mmol) were dissolved in acetic anhydride (5 mL), and the reaction solution was reacted at 90 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (5 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 4/1) to give 2- (2-fluoro-5- (6-fluoro-1H-indol-1-yl) sulfanilamide) -4-methoxybenzene) isopropyl-1,3-dione 092c (113 mg, white solid) in 74% yield.
MS m/z(ESI)：469.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.57(d,J＝8.0Hz,1H),8.03-8.05(m,2H),7.97-7.95(m,2H),7.75-7.69(m,1H),7.75-7.66(m,1H),7.45–7.41(m,2H),7.17-7.121(m,1H),6.82(d,J＝3.7Hz,1H),3.82(s,1H)。
The third step
Preparation of 2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline
2- (2-fluoro-5- (6-fluoro-1H-indol-1-yl) sulfanilamide) -4-methoxybenzene) isopropyl-1,3-dione 092c (113mg, 0.24mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL), and the reaction mixture was reacted at 90 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (5 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 4/1) to give 2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 092d (63 mg, yellow solid) in a yield of 77%.
MS m/z(ESI)：339.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ7.67-7.60(m,3H),7.40-7.37(m,1H),7.15-7.10(m,1H),7.01(d,J＝12.8Hz,1H),6.76-6.75(m,,1H),3.56(s,3H)。
The fourth step
Preparation of 4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
22-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 092d (63mg, 0.18mmol) and dimethyl 4- ((phenoxycarbonyl) amino) thienyl-2,3-dicarboxylate (124mg, 0.37mmol) were dissolved in tetrahydrofuran (5 mL), triethylamine (37mg, 0.37mmol) was added thereto, and the reaction mixture was reacted at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (5 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to give 4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 092e (63 mg, white solid) in 52% yield.
MS m/z(ESI)：580.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.31(s,1H),8.97(s,1H),8.90(d,J＝8.1Hz,1H),8.02(s,1H),7.71(d,J＝3.7Hz,1H),7.68-7.64(m,1H),7.42-7.39(m,1H),7.28(d,J＝12.7Hz,1H),7.15-7.10(m,1H),6.78(d,J＝3.7Hz,1H),3.90(s,3H),3.84(s,3H),3.67(s,3H)。
The fifth step
Preparation of 3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 092e (63mg, 0.11mmol) and lithium hydroxide (13mg, 0.54mmol) were dissolved in tetrahydrofuran: methanol: water =1:1:1 (6 mL), and the reaction mixture was reacted at room temperature for 1 hour. After completion of the reaction, the reaction mixture was quenched by addition of 1M hydrochloric acid (5 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The residue obtained was purified by HPLC preparation (mobile phase: acetonitrile/water) to give 3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-027 (7 mg, white solid) in 12% yield.
MS m/z(ESI)：534.0[M+1] + 。
HPLC:99.80％(214nm),98.95％(254nm)。
1 H NMR(400MHz,DMSO-d 6 )δ14.34(s,1H),12.07(s,1H),8.56(d,J＝8.0Hz,1H),7.76-7.66(m,2H),7.41-7.35(m,3H),7.17-7.12(m,1H),6.82(d,J＝3.7Hz,1H),3.81(s,3H)。
Example 28
Preparation of 3- (5- ((5-chloro-1H-indol-1-yl) sulfonyl) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-028)
Figure BDA0003608368340000701
First step of
Preparation of 2- [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione
5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 093a (250mg, 0.68mmol) and 5-chloro-1H-indole (123mg, 0.81mmol) were dissolved in dry THF (5 mL), sodium hydride (24mg, 1.01mmol) was added, and the reaction was carried out at 25 ℃ for 3 hours. After completion of the reaction, water (5 mL) was added to quench, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to give 2- [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 093b (184 mg, white solid) in yield: 56 percent.
MS m/z(ESI)：485.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.81(d,J＝8.0Hz,1H),7.86-7.84(m,1H),7.79(d,J＝3.6Hz,1H),7.73-7.71(m,2H),7.67(d,J＝8.8Hz,1H),7.56-7.54(m,2H),7.32(dd,J＝8.8,2.0Hz,1H),7.24(d,J＝12.2Hz,1H),6.77(d,J＝3.6Hz,1H),3.69(s,3H).
Second step of
Preparation of 5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline
2- [5- (5-Chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 093b (184mg, 0.38mmol) was dissolved in ethanol (5 mL), and hydrazine hydrate (38mg, 0.76mmol) was added to react at 90 ℃ for 1 hour. After completion of the reaction, it was quenched by addition of water (10 mL), extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 4/1) to give 5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 093c (66 mg, yellow solid) in yield: 49 percent.
MS m/z(ESI)：355.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ7.74(d,J＝3.6Hz,1H),7.71(d,J＝2.0Hz,1H),7.62(d,J＝8.8Hz,1H),7.58(d,J＝9.6Hz,1H),7.32(dd,J＝8.8,2.0Hz,1H),7.01(d,J＝12.8Hz,1H),6.73(d,J＝3.2Hz,1H),5.25(s,2H),3.54(s,3H).
The third step
Preparation of dimethyl 4- ({ [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
A solution of 5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 093c (60mg, 0.17mmol), 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dimethylester (68mg, 0.20mmol) and triethylamine (51mg, 0.51mmol) in tetrahydrofuran was stirred at 50 ℃ for 3 hours after completion of the reaction, quenched by addition of water (50 mL), extracted with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 4- ({ [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dimethylester 093d (80 mg, yellow solid) in 79% yield.
MS m/z(ESI)：596.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.32(s,1H),8.97(s,1H),8.90(d,J＝8.6Hz,1H),8.03(s,1H),7.78(d,J＝3.6Hz,1H),7.71(d,J＝2.0Hz,1H),7.64(d,J＝8.8Hz,1H),7.31(dd,J＝8.8,2.0Hz,1H),7.26(d,J＝12.8Hz,1H),6.76(d,J＝3.8Hz,1H),3.90(s,3H),3.84(s,3H),3.66(s,3H).
The fourth step
Preparation of 3- [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 093d (70mg, 0.12mmol) was dissolved in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL), and lithium hydroxide monohydrate (15mg, 0.35mmol) was added to react at 25 ℃ for 2 hours. After completion of the reaction, dilute hydrochloric acid was added to adjust pH =4-5, and extraction was performed with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-028 (10 mg, white solid) in yield: 16 percent.
MS m/z(ESI)：550.0[M+1] + 。
HPLC:99.45％(214nm),98.19％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.33(s,1H),11.99(s,1H),8.49(d,J＝8.0Hz,1H),7.81(d,J＝3.6Hz,1H),7.75(d,J＝2.0Hz,1H),7.64(d,J＝8.8Hz,1H),7.38(d,J＝11.6Hz,1H),7.32(s,1H),7.26(dd,J＝8.8,2.0Hz,1H),6.80(dd,J＝3.6,0.6Hz,1H),3.79(s,3H).
Example 29
Preparation of 3- [5- (6-Chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-029)
Figure BDA0003608368340000711
First step of
Preparation of 2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid
6-chloro-1H-indole 094a (250mg, 1.65mmol) was dissolved in tetrahydrofuran (5 mL). Sodium hydride (118mg, 4.95mmol) was then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 0.5 hour. Then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (609mg, 1.64mmol) was dissolved in tetrahydrofuran (2 mL), and the solution was added dropwise thereto, and the reaction solution was reacted at 25 ℃ for 3 hours. Then, the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, methylene chloride (2X 10 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 094b 2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid (150 mg, white solid) in 16% yield.
MS m/z(ESI)：503.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ13.18(s,1H),10.29(s,1H),8.56(d,J＝8.4Hz,1H),7.89(d,J＝7.6Hz,1H),7.72(d,J＝3.6Hz,1H),7.68(s,1H),7.64(d,J＝8.4Hz,2H),7.60-7.54(m,2H),7.30-7.21(m,2H),6.80(d,J＝3.6Hz,1H),3.67(s,3H).
Second step of
Preparation of 2- [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione
094b (150mg, 0.298mmol) 2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid was dissolved in acetic acid (2 mL). Sodium acetate (73mg, 0.895mmol) was then added. The reaction mixture was reacted at 90 ℃ for 1.5 hours. Then, the pH was adjusted to about 7 with a saturated sodium bicarbonate solution, extracted with ethyl acetate (2X 5 mL), and the organic phases were combined and dried over anhydrous sodium sulfate to give 2- [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 094c (80 mg, white solid) with a yield of 50%.
MS m/z(ESI)：485.0[M+1] + 。
The third step
Preparation of 5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline
2- [5- (6-Chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 094c (62mg, 0.13mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction mixture was reacted at 90 ℃ for 4 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 094d (35 mg, white solid) in 69% yield.
MS m/z(ESI)：355.0[M+1] + 。
The fourth step
Preparation of dimethyl 4- ({ [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 094d (30mg, 0.085 mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (34mg, 0.27mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (25mg, 0.25mmol) was added, the reaction solution was reacted at 25 ℃ for 1 hour, and the reaction solution was not subjected to any post-treatment to obtain a stock solution of 4- ({ [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 094e.
MS m/z(ESI)：596.0[M+1] + 。
The fifth step
Preparation of 3- [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 094e (5 mL stock solution) was dissolved in methanol with lithium hydroxide (6 mg, 0.14mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was finished, 3- [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-029 (2.06 mg, white solid) was obtained by direct concentration, after preliminary purification by reverse column (water: acetonitrile = 60).
MS m/z(ESI)：550.0[M+1] + 。
HPLC:97.60％(214nm),98.03％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ12.09(s,1H),8.57(d,J＝8.0Hz,1H),7.79(d,J＝3.6Hz,1H),7.68(d,J＝8.4Hz,1H),7.61(s,1H),7.42-7.36(m,2H),7.32-7.29(m,1H),6.85(d,J＝3.6Hz,1H),3.80(s,3H).
Example 30
Preparation of 3- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-030)
Figure BDA0003608368340000731
First step of
Preparation of 2- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione
5-fluoro-1H-indole 095a (60mg, 0.44mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (32mg, 1.33mmol) was added in an ice-water bath, the reaction was stirred at 0 ℃ for 15 minutes, then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (165mg, 0.44mmol) was added, and the reaction was stirred at 25 ℃ for 4 hours. The reaction was quenched with water (50 mL) and extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the title product, 2- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione 095b (200 mg, yellow oil) was obtained in 29% yield.
MS m/z(ESI)：469.1[M+1] + .
Second step of
Preparation of 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyaniline
2- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1,3-dione 095b (200mg, 0.42mmol) was dissolved in ethanol (5 mL), and hydrazine hydrate (214mg, 4.27mmol) was added. The reaction solution was stirred at 90 ℃ for 1 hour. The reaction solution was concentrated to give a residue, which was purified by a silica gel column (dichloromethane/methanol = 20/1) to give the title product 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyaniline 095c (30 mg, yellow oil) in 18% yield.
MS m/z(ESI)：339.1[M+1] + .
The third step
Preparation of 4- ({ [ [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyaniline 095c (30mg, 0.09mmol) was dissolved in tetrahydrofuran (3 mL), 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (30mg, 0.09mmol) and triethylamine (27mg, 0.27mmol) were added in this order, and the reaction was stirred at 25 ℃ for 16 hours, and the reaction was concentrated to give the title product 4- ({ [ [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 095d (40 mg, yellow oil) in 39% yield.
MS m/z(ESI)：580.1[M+1] + .
The fourth step
Preparation of 3- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- ({ [ [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 095d (30mg, 0.05mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (111mg, 0.26mmol) was added, and the reaction solution was stirred at 25 ℃ for 2 hours. Water (0.5 mL) was then added and the reaction was stirred at 25 ℃ for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution and then extracted with ethyl acetate (3X 30 mL). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was purified by preparative HPLC (acetonitrile/water (0.1% formic acid) to give the title product 3- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-030 (4.03 mg, light yellow solid) in 14% yield.
MS m/z(ESI)：534.0[M+1] + .
HPLC:98.93％(214nm),98.22％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ12.04(s,1H),8.50(d,J＝8.0Hz,1H),7.81(d,J＝3.6Hz,1H),7.64(dd,J＝9.2,4.4Hz,1H),7.48(dd,J＝9.2,2.4Hz,1H),7.38(t,J＝5.6Hz,2H),7.09(td,J＝9.2,2.4Hz,1H),6.81(d,J＝3.6Hz,1H),3.80(s,3H).
Example 31
Preparation of 3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-031)
Figure BDA0003608368340000741
First step of
Preparation of 2- ((2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid
7-fluoro-1H-indole 096a (300mg, 2.2mmol) was dissolved in a dry tetrahydrofuran (5 mL) solution and cooled to 0 ℃ under nitrogen. Sodium hydride (300mg, 11mmol) was then added and the mixture stirred for 30 min. Then 5- (1,3-dioxapolyalcohol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (410mg, 1.1mmol) was added, and the reaction solution was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was quenched with water and extracted with ethyl acetate (3X 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by chromatography column (methanol/dichloromethane = 0-10%) to give 2- { [ 2-fluoro-5- (7-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid 096b (260 mg, yellow solid) yield: 35 percent.
MS m/z(ESI)：487.1[M+1] + 。
Second step of
Preparation of 2- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) isoindole-1,3-dione
096b (160mg, 0.3mmol) 2- { [ 2-fluoro-5- (7-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid was dissolved in acetic anhydride (5 mL), followed by addition of sodium acetate (134mg, 1.6 mmol) to the solution. The reaction solution was stirred at 90 ℃ for 2 hours under nitrogen protection. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 2- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) isoindole-1,3-dione 096c (140 mg, yellow solid), yield: 70 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：491.0(M+23)。
The third step
Preparation of 2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline
2- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) isoindole-1,3-dione 096c (140mg, 0.34mmol) was dissolved in ethanol (6 mL), followed by the addition of hydrazine hydrate (68mg, 0.36mmol) to the solution. The resulting mixture was stirred at 90 ℃ for 2 hours under nitrogen. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by chromatography column (ethyl acetate/petroleum ether = 0-20%) to give 2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 096d (85 mg, brown solid) in yield: and 63 percent.
MS m/z(ESI)：339.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.78-7.75(m,,2H),7.31(d,J＝7.8Hz,1H),7.13-7.08(m,1H),6.92-6.87(m,1H),6.66-6.56(m,2H),3.62(s,3H).
The fourth step
Preparation of 4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 096d (85mg, 0.25mmol) was dissolved in tetrahydrofuran (5 mL), and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (343mg, 1.0mmol) and triethylamine (52mg, 0.51mmol) were added in this order. The reaction solution was stirred at room temperature for 16 hours. After the reaction was complete, the resulting mixture was washed with water (10 mL). The resulting mixture was then extracted with ethyl acetate (3X 10 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 096e (60 mg, brown solid), yield: 42 percent. The crude product was used directly in the next step.
MS m/z(ESI)：580.1[M+1] + 。
The fifth step
Preparation of 3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 096e (60mg, 0.10 mmol) was dissolved in tetrahydrofuran/methanol/water =1:1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography (water/acetonitrile = 40%) to give 3- [ 2-fluoro-5- (7-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-031 (30 mg, yellow solid) in yield: 53 percent.
MS m/z(ESI)：534.0[M+1] + 。
HPLC:99.13％(214nm),98.49％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ12.01(s,1H),8.42-8.39(m,1H),7.87(d,J＝3.8Hz,1H),7.48(d,J＝7.6Hz,1H),7.45-7.36(m,2H),7.25-7.20(m,1H),7.09-7.04(m,1H),6.89-6.87(m,1H),3.81(s,3H).
Example 32
Preparation of 3- [ 2-fluoro-5- (4-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-032)
Figure BDA0003608368340000751
First step of
Preparation of 2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid
4-fluoro-1H-indole 098a (250mg, 1.85mmol) was dissolved in tetrahydrofuran (5 mL) and sodium hydride (133mg, 5.55mmol) was added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 0.5 hour. Then 5- (1,3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (684mg, 1.85mmol) was dissolved in tetrahydrofuran (2 mL), and the solution was added dropwise thereto, and the reaction mixture was reacted at 25 ℃ for 3 hours. Then, the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, methylene chloride (2X 10 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 098b 2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid (230 mg, white solid) in a yield of 25%.
MS m/z(ESI)：487.0[M+1] + 。
Second step of
Preparation of 2- [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione
098b (200mg, 0.41mmol) 2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid was dissolved in acetic acid (2 mL). Then sodium acetate (101mg, 1.23mmol) was added. The reaction mixture was reacted at 90 ℃ for 1.5 hours. The pH was then adjusted to about 7 with saturated sodium bicarbonate solution, extracted with ethyl acetate (2X 5 mL), and the organic phases were combined and dried over anhydrous sodium sulfate to give 2- [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 098c (150 mg, yellow solid) in 70% yield.
MS m/z(ESI)：469.0[M+1] + 。
The third step
Preparation of 5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline
2- [5- (4-Fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1,3-dione 098c (150mg, 0.32mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction mixture was reacted at 90 ℃ for 4 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 098d (50 mg, yellow solid) in 37% yield.
MS m/z(ESI)：339.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.57(d,J＝3.6Hz,1H),7.50(d,J＝8.4Hz,1H),7.15(td,J＝8.0,5.2Hz,1H),6.87(dd,J＝9.6,8.4Hz,1H),6.76(s,1H),6.67(dd,J＝3.6,0.8Hz,1H),6.57(d,J＝12Hz,1H),3.58(s,3H).
The fourth step
Preparation of dimethyl 4- ({ [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyaniline 098d (30mg, 0.088mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (35mg, 0.10mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (26mg, 0.26mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction solution was not subjected to any post-treatment to obtain a stock solution of 4- ({ [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 098e.
MS m/z(ESI)：580.0[M+1] + 。
The fifth step
Preparation of 3- [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Adding methanol to the reaction solution in the fourth step: water =3:1 (4 mL) and lithium hydroxide (3mg, 0.078mmol). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, 3- [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-032 (2.06 mg, white solid) was obtained after direct concentration and preliminary purification by reverse column (water: acetonitrile = 60) preparation (acetonitrile/0.1% aqueous formic acid solution) purification with yield: 8%.
MS m/z(ESI)：534.0[M+1] + 。
HPLC:99.48％(214nm),98.87％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ12.04(s,1H),8.53(d,J＝8.0Hz,1H),7.80(d,J＝3.6Hz,1H),7.47(d,J＝8.4Hz,1H),7.39(d,J＝11.2Hz,2H),7.25(td,J＝8.2,5.6Hz,1H),7.10(dd,J＝9.6,8.4Hz,1H),6.89(dd,J＝3.6,0.8Hz,1H),3.81(s,3H).
Example 33
Preparation of 3- {5- [ (5,7-Difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-033)
Figure BDA0003608368340000771
First step of
Preparation of [ (2-bromo-3,5-difluorophenyl) methyl ]2,2-dimethoxyethyl) amine
2-bromo-3,5-difluorobenzaldehyde 099a (5.00g, 22.60mmol) was dissolved in ethanol (50 mL). 2,2-dimethoxyethylamine (2.61g, 24.80mmol) was then added at 25 ℃. The reaction mixture was reacted at 80 ℃ for 16 hours. The solvent was then dried by evaporation and extracted with ethyl acetate (2X 50 mL), the organic phases combined and dried over anhydrous sodium sulfate and concentrated to give [ (2-bromo-3,5-difluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 099b (5.70 g, red solid) 73% yield.
MS m/z(ESI)：310.0[M+1] + 。
Second step of
Preparation of N- [ (2-bromo-3,5-difluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine
[ (2-bromo-3,5-difluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 099b (5.70g, 1.86mmol) was dissolved in dichloromethane (60 mL). Triethylamine (5.64g, 55.80mmol) and 4-toluenesulfonyl chloride (10.64g, 55.80mmol) were then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then, it was extracted with water (2X 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give N- [ (2-bromo-3,5-difluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine 099c (7.20 g, red solid) in 75% yield.
MS m/z(ESI)：489.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.73(d,J＝8.0Hz,2H),7.34(d,J＝8.0Hz,2H),7.08(d,J＝9.2Hz,1H),6.81(td,J＝8.4,2.6Hz,1H),5.30(s,1H),4.51(s,2H),3.29(d,J＝5.2Hz,2H),3.24(s,6H),2.45(s,3H).
The third step
Preparation of 8-bromo-5,7-difluoroisoquinoline
N- [ (2-bromo-3,5-difluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine 099c (1.50g, 3.20mmol) was dissolved in dichloromethane (10 mL) and added dropwise to a solution of anhydrous aluminum trichloride (2.55g, 19.20mmol) in dichloromethane (5 mL) at 0 ℃. The reaction mixture was reacted at 25 ℃ for 16 hours. Then, the mixture was quenched by addition of ice water and extracted with water (2X 50 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give 8-bromo-5,7-difluoroisoquinoline 099d (450 mg, yellow solid) in 53% yield.
MS m/z(ESI)：244.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.73(d,J＝8.0Hz,2H),7.34(d,J＝8.0Hz,2H),7.08(d,J＝9.2Hz,1H),6.81(td,J＝8.4,2.6Hz,1H),5.30(s,1H),4.51(s,2H),3.29(d,J＝5.2Hz,2H),3.24(s,6H),2.45(s,3H).
The fourth step
Preparation of 5,7-difluoro-isoquinoline-8-carboxylic acid methyl ester
8-bromo-5,7-difluoroisoquinoline 099d (460mg, 1.88mmol) was dissolved in methanol (10 mL). Triethylamine (572mg, 5.65mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium) (137mg, 0.19mmol) were then added thereto, and the reaction solution was reacted at 80 ℃ for 16 hours under pressure sealed using a closed vessel charged with carbon monoxide. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 55/45) to give 5,7-difluoroisoquinoline-8-carboxylic acid methyl ester 099e (60 mg, brown solid) in 60% yield.
MS m/z(ESI)：224.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.70(s,1H),8.68(d,J＝5.6Hz,1H),7.91(d,J＝5.2Hz,1H),7.24(d,J＝9.6Hz,1H),4.09(s,3H).
The fifth step
Preparation of (5,7-difluoroisoquinolin-8-yl) methanol
5,7-Difluoroisoquinoline-8-carboxylic acid methyl ester 099e (100mg, 0.45mmol) was dissolved in tetrahydrofuran (5 mL). Then, lithium aluminum hydride (85mg, 2.24mmol) was added thereto at 0 ℃ and the reaction mixture was reacted at 0 ℃ for 0.5 hour. Then, the mixture was quenched by addition of ice water and extracted with dichloromethane (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give (5,7-difluoroisoquinolin-8-yl) methanol 099f (70 mg, yellow solid) in 72% yield.
MS m/z(ESI)：196.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.70(s,1H),8.65(d,J＝5.6Hz,1H),7.88(d,J＝5.6Hz,1H),7.20(t,J＝9.6Hz,1H),5.23(s,2H).
The sixth step
Preparation of 8- (bromomethyl) -5,7-difluoroisoquinoline
(5,7-Difluoroisoquinolin-8-yl) methanol 099f (70mg, 0.35mmol) was dissolved in dichloromethane (5 mL). Then, triphenylphosphine (188mg, 0.71mmol) and carbon tetrabromide (237mg, 0.71mmol) were added, and the reaction mixture was reacted at 45 ℃ for 16 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 8- (bromomethyl) -5,7-difluoroisoquinoline 099g (52 mg, brown solid) in 50% yield.
MS m/z(ESI)：258.0[M+1] + 。
Seventh step
Preparation of 5,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
8- (bromomethyl) -5,7-difluoroisoquinoline 099g (70mg, 0.27mmol) and 4-fluoro-2-methoxy-5-nitrophenol (55mg, 0.30mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (196mg, 0.28mmol) was added thereto, and the reaction mixture was reacted at 45 ℃ for 16 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 099h (14 mg, white solid) in 13% yield.
MS m/z(ESI)：365.0[M+1] + 。
Eighth step
Preparation of 5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
5,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 099h (20mg, 0.04mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (2 g) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 099i (12 mg, white solid) in 75% yield.
MS m/z(ESI)：335.0[M+1] + 。
The ninth step
Preparation of dimethyl 4- [ ({ [5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 099i (30mg, 0.10mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (42mg, 0.12mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (31mg, 0.31mmol) was added, and the reaction solution was reacted at 25 ℃ for 1 hour to obtain a stock solution 4- [ ({ [5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 099j without any post treatment.
MS m/z(ESI)：529.1[M+1] + 。
The tenth step
Preparation of 3- {5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Prepare for
4- [ ({ [5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 099j (5 mL stock solution) and lithium hydroxide (3mg, 0.078mmol) were dissolved in methanol: water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, 3- {5- [ (5,7-difluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-033 (1.11 mg, white solid) was obtained by direct concentration after preparative purification by reverse column (water: acetonitrile = 82).
MS m/z(ESI)：530.0[M+1] + 。
HPLC:99.40％(214nm),97.26％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.85(s,1H),9.62(s,1H),8.65(d,J＝5.6Hz,1H),7.96(d,J＝5.6Hz,1H),7.83(t,J＝10.2Hz,1H),7.32(d,J＝7.2Hz,1H),7.21(s,1H),7.13(d,J＝11.6Hz,1H),5.49(s,2H),3.76(s,3H).
Example 34
Preparation of 3- {5- [ (1-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-034)
Figure BDA0003608368340000791
First step of
Preparation of isoquinoline-8-carboxylic acid methyl ester
8-Bromoisoquinoline 100a (1.80g, 8.65mmol) was dissolved in methanol (30 mL), followed by addition of Pd (dppf) Cl 2 (633mg, 0.87mmol) and triethylamine (2.63g, 25.95mmol). The reaction solution was stirred at 80 ℃ for 16 hours under carbon monoxide protection. Upon completion of the LCMS detection reaction, the organic solvent was removed by distillation under reduced pressure and then subjected to silica gel column (petroleum ether/ethyl acetate = 6/4) to obtain the target product methyl isoquinoline-8-carboxylate 100b (1.00 g, white solid) in yield: 56 percent.
MS m/z(ESI)：188.1[M+1] + 。
Second step of
Preparation of 2-oxo-8- (methoxycarbonyl) isoquinoline
Isoquinoline-8-carboxylic acid methyl ester 100b (1.50g, 8.01mmol) was dissolved in chloroform (10 mL) and then m-chloroperoxybenzoic acid (2.77g, 16.03mmol) was added while cooling on ice. Stir at rt for 3h. After the reaction was completed, a small amount of saturated solution of sodium carbonate was added, the sodium carbonate solution layer was extracted with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to remove the solvent to obtain the target product 2-oxo-8- (methoxycarbonyl) isoquinoline 100c (600 mg, brown solid), yield: 33 percent.
MS m/z(ESI)：204.0[M+1] + 。
The third step
Preparation of 1-chloroisoquinoline-8-carboxylic acid methyl ester
2-Oxo-8- (methoxycarbonyl) isoquinoline 100c (450mg, 2.21mmol) was dissolved in phosphorus oxychloride (8 mL). The reaction was stirred at 105 ℃ for 4h. After completion of the reaction, the solvent was drained, saturated sodium carbonate solution was added to adjust the pH to 7-8, followed by extraction with dichloromethane (3X 20 mL), drying over anhydrous sodium sulfate, and concentration under reduced pressure to give methyl 1-chloroisoquinoline-8-carboxylate 100d (200 mg, brown oil) in yield: 37 percent.
MS m/z(ESI)：222.2[M+1] + 。
The fourth step
Preparation of (1-chloroisoquinolin-8-yl) methanol
Methyl 1-chloroisoquinoline-8-carboxylate 100d (300mg, 1.35mmol) was dissolved in tetrahydrofuran (20 mL) and diisobutylaluminum hydride (10.8mL, 10.83mmol,1M in n-hexane) was added under ice-bath. The reaction solution was stirred at 0 ℃ for 1 hour. After completion of the reaction, a saturated ammonium chloride solution was added, followed by extraction with dichloromethane (3 × 20 mL), washing of the organic phase with saturated brine, drying over anhydrous sodium sulfate, filtration and concentration under reduced pressure to remove the solvent, and purification by silica gel column (petroleum ether/ethyl acetate = 1/1) gave (1-chloroisoquinolin-8-yl) methanol 100e (120 mg, white solid) in yield: 44 percent.
MS m/z(ESI)：194.2[M+1] + 。
The fifth step
Preparation of 8- (bromomethyl) -1-chloroisoquinoline
(1-Chloroisoquinolin-8-yl) methanol 100e (100mg, 0.52mmol) as solvent in dichloromethane (10 mL) was added, followed by carbon tetrabromide (205mg, 0.62mmol) and triphenylphosphine (163mg, 0.62mmol). The reaction was stirred at room temperature for 1 h. After completion of the reaction, the solvent was concentrated by evaporation to give a crude product, which was concentrated off the solvent after passing through a silica gel column (petroleum ether/ethyl acetate = 6/4) to give 8- (bromomethyl) -1-chloroisoquinoline 100f (120 mg, white solid) in yield: 82 percent.
MS m/z(ESI)：256.0[M+1] + 。
The sixth step
Preparation of 1-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
8- (bromomethyl) -1-chloroisoquinoline 100f (100mg, 0.39mmol) was dissolved in acetonitrile (20 mL), followed by the addition of 4-4-fluoro-2-methoxy-5-nitrophenol (80mg, 0.43mmol), potassium carbonate (108mg, 0.78mmol) and potassium iodide (6mg, 0.04mmol). The reaction was carried out at 65 ℃ for 2 hours. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent, and the crude product was purified by silica gel column (petroleum ether/ethyl acetate = 7/3) to give 100g (150 mg, pale yellow solid) of 1-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline, yield: 85 percent.
MS m/z(ESI)：363.2[M+1] + 。
Seventh step
Preparation of 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
100g (120mg, 0.33mmol) of 1-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (185mg, 3.31mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and extracted with dichloromethane (3X 20 mL) and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 100h (100 mg, light pink solid) in yield: 73 percent.
MS m/z(ESI)：333.1[M+1] + 。
Eighth step
Preparation of 4- [ ({ 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
5- [ (1-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline (50mg, 0.15mmol) was dissolved in tetrahydrofuran (10 mL) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (60mg, 0.18mmol) and triethylamine (46mg, 0.45mmol) were added. The reaction solution was stirred at 25 ℃ for 16 hours. After completion of the reaction monitored by LCMS, the compound 4- [ ({ 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 100i (100 mg, light yellow liquid) was obtained after the reaction, yield: 54 percent.
MS m/z(ESI)：574.1[M+1] + 。
The ninth step
Preparation of 3- {5- [ (1-Chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction mixture in the seventh step. Lithium hydroxide monohydrate (11mg, 0.26mmol) was added. The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give the crude product which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- {5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-034 (20 mg, light yellow solid), yield: 42 percent.
MS m/z(ESI)：528.1[M+1] + 。
HPLC:97.21％(214nm),97.20％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.52(s,1H),11.95(s,1H),8.29(d,J＝5.4Hz,1H),8.08(d,J＝7.6Hz,1H),7.98(d,J＝7.2Hz,1H),7.93(d,J＝5.4Hz,1H),7.87-7.82(m,1H),7.35(s,1H),7.29(d,J＝7.2Hz,1H),7.15(d,J＝11.4Hz,1H),5.72(s,2H),3.82(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-128.48.
Example 35
Preparation of 3- (2-fluoro-5- ((6-fluoro-2,3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) methyl) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-035)
Figure BDA0003608368340000811
First step of
Preparation of 6-fluoro-3,4-dihydro-2H-1,4-benzoxazine
To a solution of 2-amino-4-fluorophenol 102a (1.00g, 7.90mmol) and potassium carbonate (2.18g, 15.80mmol) in N, N-dimethylformamide (10 mL) was added 1,2-dibromoethane (1.78g, 9.40mmol), and the mixture was reacted at 80 ℃ for 8 hours. After the reaction was completed, quenching was performed by adding water (10 mL), extraction was performed with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 6-fluoro-3,4-dihydro-2H-1,4-benzoxazine 102b (178 mg, yellow oil), yield: 15 percent.
MS m/z(ESI)：154.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ6.60(dd,J＝8.6,5.6Hz,1H),6.34(dd,J＝10.6,3.0Hz,1H),6.20(td,J＝8.6,3.0Hz,1H),6.05(s,1H),4.09-4.03(m,2H),3.29-3.24(m,2H).
Second step of
Preparation of 6-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine
To a solution of 6-fluoro-3,4-dihydro-2H-1,4-benzoxazine 102b (90mg, 0.59mmol) and potassium carbonate (122mg, 0.88mmol) in acetonitrile (5 mL) was added 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (155mg, 0.59mmol) and reacted at 70 ℃ for 8 hours. After completion of the reaction, quenching was performed by addition of water (5 mL), extraction was performed with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to give 6-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 102c (190 mg, yellow solid), yield: 96 percent.
MS m/z(ESI)：337.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ7.88(d,J＝8.8Hz,1H),7.33(d,J＝13.6Hz,1H),6.70(dd,J＝8.8,5.8Hz,1H),6.42(dd,J＝11.4,2.8Hz,1H),6.31(td,J＝8.4,2.8Hz,1H),4.43(s,2H),4.19-4.16(m,2H),3.98(s,3H),3.43-3.40(m,2H).
The third step
Preparation of 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline
To a solution of 6-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 102c (190mg, 0.57mmol) in ethanol (5 mL) and water (5 mL) were added iron powder (95mg, 1.69mmol) and ammonium chloride (91mg, 1.69mmol), and the mixture was reacted at 80 ℃ for 1 hour. After the reaction was completed, ethyl acetate was added for extraction (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to give 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline 102d (124 mg, yellow oil) in yield: 72 percent.
MS m/z(ESI)：307.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ6.70(dd,J＝8.4,5.6Hz,1H),6.63(d,J＝12.0Hz,2H),6.31-6.24(m,2H),4.31(s,2H),4.23-4.21(m,2H),3.78(s,3H),3.41-3.39(m,2H).
The fourth step
Preparation of dimethyl 4- [ ({ 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
To a solution of 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline 102d (114mg, 0.37mmol) and triethylamine (56mg, 0.56mmol) in tetrahydrofuran (5 mL) was added 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (150mg, 0.45mmol) and reacted at 25 ℃ for 3 hours, after completion of the reaction, water quenching was added and extraction with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl acetate = 3/1) to give 4- [ ({ { 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3 dimethyl ester as a white solid in a yield of 4398 mg, 4398%.
MS m/z(ESI)：548.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.89(s,1H),8.76(s,1H),7.85(s,1H),7.75(d,J＝9.2Hz,1H),7.05(d,J＝12.8Hz,1H),6.66(dd,J＝8.6,5.8Hz,1H),6.35(dd,J＝11.6,2.8Hz,1H),6.25(td,J＝8.6,2.8Hz,1H),4.36(s,2H),4.18-4.16(m,2H),3.85(s,3H),3.82-3.81(m,6H),3.42-3.39(m,2H).
The fifth step
Preparation of 3- { 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Lithium hydroxide monohydrate (23mg, 0.55mmol) was added to a solution of dimethyl 4- [ ({ { 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 102e (100mg, 0.18mmol) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL), and stirred at 25 ℃ for 2 hours. After completion of the reaction, dilute hydrochloric acid was added to adjust pH =4-5, and extraction was performed with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by reverse phase column chromatography (mobile phase: 45% acetonitrile/55% water) to give 3- { 2-fluoro-5- [ (6-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-035 (75 mg, yellow solid), yield: 82 percent.
MS m/z(ESI)：502.0[M+1] + 。
HPLC:99.07％(214nm),96.70％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.10-7.06(m,2H),6.63(dd,J＝8.6,5.8Hz,1H),6.51(s,1H),6.44(dd,J＝11.6,2.8Hz,1H),6.25(td,J＝8.4,2.8Hz,1H),4.37(s,2H),4.15-4.12(m,2H),3.88(s,3H),3.47-3.39(m,2H).
Example 36
Preparation of 3- (2-fluoro-5- ((5-fluoro-2,3-dihydro-4H-benzo [ b ] [1,4] oxazin-4-yl) methyl) -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-036)
Figure BDA0003608368340000831
First step of
Preparation of 5-fluoro-3,4-dihydro-2H-1,4-benzoxazine
To a solution of 2-amino-3-fluorophenol 103a (1.00g, 7.90mmol) and potassium carbonate (2.18g, 15.80mmol) in N, N-dimethylformamide (10 mL) was added 1,2-dibromoethane (1.78g, 9.40mmol), and the reaction was carried out at 80 ℃ for 8 hours. After completion of the reaction, water (10 mL) was added for quenching, extraction was performed with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 5-fluoro-3,4-dihydro-2H-1,4-benzoxazine 103b (262 mg, yellow oil), yield: 22 percent.
MS m/z(ESI)：154.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ6.65-6.60(m,1H),6.54-6.51(m,1H),6.48-6.42(m,1H),5.64(s,1H),4.15-4.12(m,2H),3.29-3.27(m,2H).
Second step of
Preparation of 5-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine
To a solution of 5-fluoro-3,4-dihydro-2H-1,4-benzoxazine 103b (90mg, 0.59mmol) and potassium carbonate (122mg, 0.88mmol) in acetonitrile (5 mL) was added 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (155mg, 0.59mmol) and reacted at 70 ℃ for 8 hours. After completion of the reaction, water (5 mL) was added for quenching, extraction with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give 5-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 103c (181 mg, yellow solid) in yield: 92 percent.
MS m/z(ESI)：337.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.24(d,J＝8.8Hz,1H),7.30(d,J＝13.6Hz,1H),6.83-6.78(m,1H),6.75-6.67(m,2H),4.21(s,2H),4.09-4.07(m,2H),3.91(s,3H),3.12-3.10(m,2H).
The third step
Preparation of 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline
To a solution of 5-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 103c (181mg, 0.54mmol) in ethanol (5 mL) and water (5 mL) were added iron powder (90mg, 1.61mmol) and ammonium chloride (86mg, 1.61mmol), and the mixture was reacted at 80 ℃ for 1 hour. After the reaction was completed, ethyl acetate was added for extraction (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to obtain 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline 103d (150 mg, yellow oil), yield: 91 percent.
MS m/z(ESI)：307.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.04(d,J＝10.0Hz,1H),6.80-6.74(m,1H),6.67-6.59(m,3H),4.19(s,2H),4.08-4.05(m,2H),3.71(s,3H),3.08-3.06(m,2H).
The fourth step
Preparation of dimethyl 4- [ ({ { 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
To a solution of 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyaniline 103d (140mg, 0.46mmol) and triethylamine (56mg, 0.56mmol) in tetrahydrofuran (5 mL) was added 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (184mg, 0.55mmol), reacted at 25 ℃ for 3 hours, after completion of the reaction, water quenched and extracted with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl acetate = 3/1) to give 4- [ ({ { 2-fluoro-5- [ (5-fluoro-2,3-dihydro-3562 zxft-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3 dimethyl ester as a white solid in yield (4388 mg ).
MS m/z(ESI)：548.0[M+1] + 。
1 H NMR(400MHz,DMSO-d6)δ8.93(s,1H),8.79(s,1H),8.06(d,J＝9.4Hz,1H),7.91(s,1H),7.01(d,J＝13.0Hz,1H),6.80-6.68(m,2H),6.66-6.64(m,1H),4.18(s,2H),4.04-4.02(m,2H),3.88(s,3H),3.82(s,3H),3.73(s,3H),3.05-3.03(m,2H).
Five steps
Preparation of 3- { 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Lithium hydroxide monohydrate (23mg, 0.55mmol) was added to a solution of dimethyl 4- [ ({ { 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 103e (100mg, 0.18mmol) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL), and stirred at 25 ℃ for 2 hours. After completion of the reaction, dilute hydrochloric acid was added to adjust pH =4-5, and extraction was performed with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by reverse phase column chromatography (mobile phase: 45% acetonitrile/55% water) to give 3- { 2-fluoro-5- [ (5-fluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-036 (74 mg, white solid), yield: 81 percent.
MS m/z(ESI)：502.0[M+1] + 。
HPLC:96.22％(214nm),97.04％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.42(d,J＝8.8Hz,1H),7.08(d,J＝11.8Hz,1H),6.80-6.63(m,4H),4.20(s,2H),4.04-4.00(m,2H),3.78(s,3H),3.05-3.04(m,2H).
Example 37
Preparation of 3- [5- (2,3-dihydro-1,4-benzoxazin-4-ylmethyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-037)
Figure BDA0003608368340000851
First step of
Preparation of 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene
(4-fluoro-2-methoxy-5-nitrophenyl) methanol 104a (60mg, 0.30mmol), triphenylphosphine (156mg, 0.60mmol) and carbon tetrabromide (198mg, 0.60mmol) were dissolved in dichloromethane (5 mL). The mixture was stirred at 25 ℃ for 2 hours and then concentrated, and the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene 104b (50 mg, white solid) in yield: 56 percent.
MS m/z(ESI)：264.0[M+1] + 。
Second step of
Preparation of 4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine
1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene 104b (50mg, 0.19mmol), 3,4-dihydro-2H-1,4-benzoxazine (51mg, 0.38mmol) and triethylamine (61mg, 1.51mmol) were dissolved in acetonitrile (5 mL), the mixture was stirred at 90 ℃ for 1 hour and concentrated, and the resulting residue was separated and purified with a silica gel column (petroleum ether/ethyl acetate = 1/2) to give 4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 104c (35 mg, orange solid) in yield: 46 percent.
MS m/z(ESI)：319.1[M+1] + 。
The third step
Preparation of 5- (2,3-dihydro-1,4-benzoxazin-4-ylmethyl) -2-fluoro-4-methoxyaniline
4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl]-2,3-dihydro-1,4-benzoxazine 104c (35mg, 0.11mmol) dissolved in NH 4 To a Cl/MeCN (4 mL) solution, iron powder (308mg, 5.50mmol) was added, and the mixture was stirred at 25 ℃ for 1 hour. The reaction solution is filtered andconcentration gave 5- (2,3-dihydro-1,4-benzoxazine-4-methyl) -2-fluoro-4-methoxyaniline 104d (25 mg, brown oil). The yield was 47%.
MS m/z(ESI)：289.1[M+1] + 。
The fourth step
Preparation of dimethyl 4- ({ [ (5- (2,3-dihydro-1,4-benzoxazin-4-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (2,3-dihydro-1,4-benzoxazine-4-methyl) -2-fluoro-4-methoxyaniline 104d (25mg, 0.09mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (145mg, 0.43mmol) were dissolved in tetrahydrofuran (5 mL) then triethylamine (48mg, 0.87mmol) was added and the mixture was stirred at 25 ℃ for 16 h the reaction solution was not post-treated to give a stock solution of 4- ({ [ (5- (2,3-dihydro-1,4-benzoxazine-4-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate prepared 104e (bright yellow solution) which was used directly in the next reaction.
MS m/z(ESI)：530.1[M+1] + 。
The fifth step
Preparation of 3- [5- (2,3-dihydro-1,4-benzoxazin-4-ylmethyl) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution in the fourth step, methanol (3 mL) and water (1 mL) were added, followed by lithium hydroxide (9 mg, 0.38mmol). The mixture was reacted at 25 ℃ for 90 minutes. After the reaction, the reaction mixture was directly concentrated, primarily purified by a reverse column (water: acetonitrile = 3:2), and then purified by a preparation (acetonitrile/0.1% formic acid aqueous solution) to obtain 3- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid preparation Cpd-037 (2.10 mg, orange solid) with a yield of 5%.
MS m/z(ESI)：484.1[M+1] + 。
HPLC:99.42％(214nm),97.83％(254nm)。
1 H NMR(400MHz,d 6- DMSO)11.88(s,1H),7.32-7.29(m,2H),7.16(d,J＝12.0Hz,1H),6.69-6.64(m,2H),6.53-6.47(m,2H),4.37(s,2H),4.21-4.19(m,2H),3.91(s,3H),3.38-3.36(m,2H).
Example 38
Preparation of 4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -3- (methoxycarbonyl) thiophene-2-carboxylic acid (Cpd-038)
Figure BDA0003608368340000861
First step of
Preparation of 2,3-difluoro-6-methoxyaniline
2,3-difluoro-6-methoxybenzoic acid 105a (700mg, 3.72mmol) was dissolved in N, N-dimethylformamide (20 mL). Triethylamine (565mg, 5.58mmol) and diphenylphosphorylazide (565mg, 5.58mmol) were added in this order. The reaction solution was reacted at room temperature for 3 hours. After the reaction was completed, pH =7-8 was adjusted with saturated sodium bicarbonate solution, followed by extraction with ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give 2,3-difluoro-6-methoxyaniline 105b (640 mg, yellow solid) in 97% yield.
MS m/z(ESI)：160.2[M+1] + 。
Second step of
Preparation of 2-amino-3,4-difluorophenol
2,3-difluoro-6-methoxyaniline 105b (580mg, 3.64mmol) was dissolved in dichloromethane (5 mL), and boron tribromide (4.57g, 18.22mmol) was added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then, the pH =7-8 was adjusted with saturated sodium bicarbonate, extracted with ethyl acetate (2 × 5 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 2-amino-3,4-difluorophenol 105c (550 mg, brown oily liquid) in 65% yield.
MS m/z(ESI)：146.2[M+1] + 。
The third step
Preparation of 5,6-difluoro-3,4-dihydro-2H-1,4-benzoxazine
2-amino-3,4-difluorophenol 105c (500mg, 3.45mmol) was dissolved in N, N-dimethylformamide (5 mL). Potassium carbonate (2.59g, 10.34mmol) and 1,2-dibromoethane (777mg, 4.13mmol) were then added in this order, and the reaction mixture was reacted at 85 ℃ for 16 hours. After completion of the reaction, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5,6-difluoro-3,4-dihydro-2H-1,4-benzoxazine 105d (246 mg, brown oily liquid) in 33% yield.
MS m/z(ESI)：172.2[M+1] + 。
The fourth step
Preparation of 5,6-difluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine
5,6-difluoro-3,4-dihydro-2H-1,4-benzoxazine 105d (120mg, 0.70mmol) and 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (222mg, 0.84mmol) were dissolved in N, N-dimethylformamide (5 mL). Then, potassium carbonate (290mg, 2.10mmol) and potassium iodide (8mg, 0.004mmol) were added, and the reaction solution was reacted at 65 ℃ for 16 hours. After completion of the reaction, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petrol ether/ethyl acetate: 80/20) to give 5,6-difluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 105e (150 mg, brown oily liquid) in 54% yield.
MS m/z(ESI)：355.1[M+1] + 。
The fifth step
Preparation of 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyaniline
5,6-difluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2,3-dihydro-1,4-benzoxazine 105e (120mg, 0.34mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (189mg, 3.39mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyaniline 105f (140 mg, brown oily liquid) in 95% yield.
MS m/z(ESI)：325.2[M+1] + 。
The sixth step
Preparation of dimethyl 4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-2,3-dicarboxylate
5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyaniline 105f (60mg, 0.19mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (124mg, 0.37mmol) were dissolved in tetrahydrofuran (5 mL). Then, triethylamine (56mg, 0.56mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction solution was not subjected to any post-treatment to give 109g of dimethyl 4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-2,3-dicarboxylate as a starting solution.
MS m/z(ESI)：566.1[M+1] + 。
Seventh step
Preparation of 4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -3- (methoxycarbonyl) thiophene-2-carboxylic acid
4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 109g (5 mL stock solution) was dissolved in lithium hydroxide (13mg, 0.53mmol) in methanol water =3:1 (4 mL.) the reaction solution was reacted at 25 ℃ for 90 minutes after completion of the reaction, after preliminary purification by reverse column (water: acetonitrile = 30) direct concentration to yield 4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -3- (methoxycarbonyl) thiophene-2-carboxylic acid d-038 (13.24 mg, 13.13 mg) yield of yellow solid.
MS m/z(ESI)：520.2[M+1] + 。
HPLC:99.22％(214nm),99.69％(254nm).
1 H NMR(400MHz,CDCl 3 )δ14.37(s,1H),10.16(s,1H),7.49(d,J＝8.4Hz,1H),6.90(s,1H),6.84(d,J＝11.2Hz,1H),6.62-6.51(m,2H),4.42-4.28(m,2H),4.14-4.00(m,2H),3.89(s,3H),3.22-3.08(m,2H).
Example 39
Preparation of 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-039)
Figure BDA0003608368340000881
First step of
Preparation of isoquinolin-8-yl methanol
Isoquinoline-8-carbaldehyde 107a (300mg, 1.91mmol) was dissolved in anhydrous methanol (10 mL) and then sodium borohydride (22mg, 0.57mmol) was added under an ice bath. The reaction was stirred at 0 ℃ for 1 hour. Upon completion of the reaction by LCMS, saturated ammonium chloride solution (2 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate, and the organic solvent was distilled off under reduced pressure to give isoquinolin-8-ylcarbinol 107b (250 mg, yellow oil), yield: 74 percent.
MS m/z(ESI)：160.1[M+1] + 。
Second step of
Preparation of 8- (bromomethyl) isoquinoline
Isoquinolin-8-ylcarbinol 107b (300mg, 1.88mmol) was dissolved in tetrahydrofuran (5 mL), followed by the addition of boron tribromide (763mg, 2.82mmol) while cooling on ice. Stir at rt for 1h. After completion of the reaction, the reaction mixture was poured into ice, followed by addition of a sodium carbonate solution to adjust the pH to 6-7, followed by extraction with methylene chloride (3X 10 mL), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. After concentration under reduced pressure and removal of the solvent, the objective product 8- (bromomethyl) isoquinoline 107c (180 mg, yellow oil) was obtained in yield: 34 percent.
MS m/z(ESI)：222.2[M+1] + 。
The third step
Preparation of 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline
8- (bromomethyl) isoquinoline 107c (150mg, 0.67mmol) and 4-fluoro-2-methoxy-5-nitroaniline (188mg, 1.01mmol) were dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (187mg, 1.35mmol). The reaction was stirred at 50 ℃ for 2h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), and after drying over anhydrous sodium sulfate, the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 1/2) to give 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline 107d (100 mg, yellow solid), yield: 41 percent.
MS m/z(ESI)：328.1[M+1] + 。
The fourth step
Preparation of 4-fluoro-6-methoxy-1-N- (quinolin-8-ylmethyl) benzene-1,3-diamine
Preparation 107d (50mg, 0.15mmol) of 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (85mg, 1.53mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and then extracted with dichloromethane (3X 20 mL) and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4-fluoro-6-methoxy-1-N- (quinolin-8-ylmethyl) benzene-1,3-diamine 107e (40 mg, light yellow oil), yield: 84 percent.
MS m/z(ESI)：298.1[M+1] + 。
The fifth step
Preparation of 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide 111e (40mg, 0.13mmol) was dissolved in tetrahydrofuran (10 mL) followed by the addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (54mg, 0.16mmol) and triethylamine (41mg, 0.31mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. Upon completion of the reaction monitored by LCMS, compound 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 1107f (80 mg, light yellow liquid) was obtained, yield: 66 percent.
MS m/z(ESI)：539.1[M+1] + 。
The sixth step
Preparation of 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction mixture in the fifth step. Lithium hydroxide monohydrate (9 mg, 0.22mmol) was added. The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give a crude product which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-039 (20 mg, yellow solid) in yield: 54 percent.
MS m/z(ESI)：493.2[M+1] + 。
HPLC:99.81％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.68(s,1H),11.86(s,1H),9.61(s,1H),8.52(d,J＝5.6Hz,1H),7.83(t,J＝6.0Hz,2H),7.71-7.64(m,1H),7.58(d,J＝7.2Hz,1H),7.30(s,1H),6.99(d,J＝11.6Hz,1H),6.62(d,J＝7.6Hz,1H),5.72(t,J＝6.0Hz,1H),4.86(d,J＝6.0Hz,2H),3.90(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-135.56.
Example 40
Preparation of 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } -2,4-dioxo-1H-thiophene [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-040)
Figure BDA0003608368340000891
First step of
Preparation of 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-N-methyl-5-nitroaniline
4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline 108a (40mg, 0.12mmol) was dissolved in methanol (10 mL), followed by addition of acetic acid (0.05 mL), paraformaldehyde (18mg, 0.61mmol), stirring of the mixture at room temperature for 30 min, and finally addition of sodium cyanoborohydride (23mg, 0.37mmol). The reaction was stirred at 55 ℃ for 48h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), and after drying over anhydrous sodium sulfate, the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 1/5) to give 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-N-methyl-5-nitroaniline 108b (20 mg, yellow solid), yield: and 43 percent.
MS m/z(ESI)：342.1[M+1] + 。
Second step of
Preparation of 4-fluoro-2-methoxy-N-methyl-5-nitro-N- (isoquinolin-8-ylmethyl) aniline
4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-N-methyl-5-nitroaniline 108b (50mg, 0.15mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (82mg, 1.46mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and extracted with dichloromethane (3X 20 mL) and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4-fluoro-2-methoxy-N-methyl-5-nitro-N- (isoquinolin-8-ylmethyl) aniline 108c (40 mg, light yellow oil) as a yield: 79 percent.
MS m/z(ESI)：312.1[M+1] + 。
The third step
Preparation of 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
4-fluoro-2-methoxy-N-methyl-5-nitro-N- (quinolin-8-ylmethyl) aniline 108c (30mg, 0.10mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (39mg, 0.12mmol) and triethylamine (29mg, 0.29mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. Upon completion of the reaction monitored by LCMS, compound 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 108d (60 mg, light yellow liquid) was obtained in yield: 56 percent.
MS m/z(ESI)：553.1[M+1] + 。
The fourth step
Preparation of 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } -2,4-dioxo-1H-thiophene [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction mixture in the third step. Lithium hydroxide monohydrate (7mg, 0.16mmol) was added. The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give the crude product, which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } -2,4-dioxo-1H-thiophene [3,4-d ] pyrimidine-5-carboxylic acid Cpd-040 (10 mg, light yellow solid), yield: 21 percent.
MS m/z(ESI)：507.1[M+1] + 。
HPLC:100％(214nm),99.65％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.92(s,1H),9.85(s,1H),8.52(d,J＝5.6Hz,1H),7.90(d,J＝7.2Hz,1H),7.83(d,J＝5.6Hz,1H),7.76–7.67(m,2H),7.32(s,1H),7.23(d,J＝8.0Hz,1H),7.13(d,J＝11.6Hz,1H),5.76(s,2H),3.90(s,3H),3.36(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-127.13–-127.37.
EXAMPLE 41
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (quinoline-8-methyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-041)
Figure BDA0003608368340000911
First step of
Preparation of quinolin-8-yl-methanols
Quinoline 8-carbaldehyde 109a (300mg, 1.9 mmol) was dissolved in methanol (5 mL). Sodium borohydride (42mg, 0.19mmol) was then added at 0 deg.C. The reaction mixture was reacted at 0 ℃ for 1 hour. Then, dichloromethane (2X 10 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated to give quinolin-8-yl-methanol 109b (190 mg, yellow solid) in 96% yield.
MS m/z(ESI)：160.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.89(dd,J＝4.4,1.6Hz,1H),8.23(dd,J＝8.4,1.6Hz,1H),7.78(dd,J＝8.4,1.2Hz,1H),7.63-7.58(m,1H),7.48-7.52(m,2H),5.22(s,2H).
Second step of
Preparation of 8- (bromomethyl) quinoline
Quinolin-8-ylmethanol 109b (100mg, 0.63mmol) was dissolved in dichloromethane (5 mL). Triphenylphosphine (329mg, 1.2mmol) and carbon tetrabromide (416mg, 1.2mmol) were then added. The reaction mixture was reacted at 25 ℃ for 1 hour. Then, dichloromethane (2X 5 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 8- (bromomethyl) quinoline 109c (100 mg, red solid) in 52% yield.
MS m/z(ESI)：224.0[M+1] + 。
The third step
Preparation of 4-fluoro-2-methoxy-5-nitro-N- (quinoline-8-methyl) aniline
8- (bromomethyl) quinoline 109c (100mg, 0.47mmol) and 4-fluoro-2-methoxy-5-nitrophenol (96mg, 0.52mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (196mg, 1.52mmol) and potassium iodide (8mg, 0.004mmol) were added thereto, and the reaction solution was reacted at 25 ℃ for 1 hour. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 50/50) to give 4-fluoro-2-methoxy-5-nitro-N- (quinoline-8-methyl) aniline 109d (80 mg, yellow solid) in 46% yield.
MS m/z(ESI)：328.1[M+1] + 。
The fourth step
Preparation of 4-fluoro-6-methoxy-1-N- (quinoline-8-methyl) benzene-1,3-diamine
4-fluoro-2-methoxy-5-nitro-N- (quinolin-8-methyl) aniline 109d (80mg, 0.24mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (2 g) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 4-fluoro-6-methoxy-1-N- (quinolin-8-methyl) benzene-1,3-diamine 109e (60 mg, white solid) in 74% yield.
MS m/z(ESI)：287.1[M+1] + 。
The fifth step
Preparation of dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoline-8-methyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
4-fluoro-6-methoxy-1-N- (quinoline-8-methyl) benzene-1,3-diamine 109e (30mg, 0.10mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (37mg, 0.111mmol) were dissolved in tetrahydrofuran (5 mL), and then triethylamine (30mg, 0.30mmol) was added, and the reaction solution was reacted at 25 ℃ for 1 hour to obtain a raw solution of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoline-8-methyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 109f without any post-treatment.
MS m/z(ESI)：539.1[M+1] + 。
The sixth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (quinolin-8-methyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (quinoline-8-methyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 109f (5 mL stock) was dissolved in methanol with lithium hydroxide (3mg, 0.078mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was finished, the reaction was directly concentrated and purified by preparative (acetonitrile/0.1% aqueous formic acid) preliminary purification on a reverse column (water: acetonitrile = 82) to give 3- { 2-fluoro-4-methoxy-5- [ (quinoline-8-methyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-041 (2.07 mg, yellow solid) in 7% yield.
MS m/z(ESI)：493.1[M+1] + 。
HPLC:98.23％(214nm),96.87％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.76(s,1H),8.94(dd,J＝4.2,1.6Hz,1H),8.37(dd,J＝8.4,1.6Hz,1H),7.87(d,J＝7.2Hz,1H),7.69(d,J＝6.4Hz,1H),7.58-7.51(m,2H),7.21(s,1H),6.96(d,J＝11.6Hz,1H),6.61(d,J＝7.6Hz,1H),4.86(d,J＝6.0Hz,2H),3.87(s,3H).
Example 42
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-042)
Figure BDA0003608368340000921
First step of
Preparation of 4-fluoro-2-methoxy-N- (naphthalen-1-ylmethyl) -5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline 110a (300mg, 1.61mmol), 2- (chloromethyl) naphthalene (313mg, 1.77mmol), potassium iodide (134mg, 0.81mmol), potassium carbonate (445mg, 3.22mmol) were dissolved in acetonitrile (10 mL). The mixture was reacted at 80 ℃ for 4 hours, then extracted with dichloromethane (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain preparation 110b (400 mg, orange solid) of 4-fluoro-2-methoxy-N- (naphthalen-1-ylmethyl) -5-nitroaniline, yield: 61 percent.
MS m/z(ESI)：349.1(M+23)。
1 H NMR(400MHz,CDCl 3 )δ8.04-8.01(m,1H),7.93-7.91(m,1H),7.86(d,J＝8.0Hz,1H),7.56-7.44(m,4H),7.35(d,J＝7.2Hz,1H),6.63(d,J＝12.4Hz,1H),4.76(s,2H),3.88(s,3H).
Second step of
Preparation of 4-fluoro-6-methoxy-1-N- (naphthalen-1-ylmethyl) benzene-1,3-diamine
4-fluoro-2-methoxy-N- (naphthalen-1-ylmethyl) -5-nitroaniline 110b (200mg, 0.61mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (686mg, 12.26mmol) and the mixture was stirred at 25 ℃ for 1 hour. The reaction was filtered and concentrated to give preparation 110c (120 mg, brown solid) of 4-fluoro-6-methoxy-1-N- (naphthalen-1-ylmethyl) benzene-1,3-diamine in 46% yield.
MS m/z(ESI)：297.1[M+1] + 。
The third step
Preparation of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
Preparation of 4-fluoro-6-methoxy-1-N- (naphthalen-1-ylmethyl) benzene-1,3-diamine 110c (60mg, 0.20mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (339mg, 1.01mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (113mg, 2.02mmol) was added, the mixture was stirred at 25 ℃ for 16 hours, the reaction solution was not subjected to any post-treatment to obtain a stock of dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 110d (bright yellow solution) which was used directly in the next reaction.
MS m/z(ESI)：538.1[M+1] + 。
The fourth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 110d (5 mL stock) was dissolved in methanol with lithium hydroxide (1695g, 0.65mmol): water =3:1 (4 mL). The mixture was reacted at 25 ℃ for 1 hour. After the reaction was completed, the reaction mixture was directly concentrated and preliminarily purified by reverse column (water: acetonitrile = 3:2) to prepare 3- { 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-042 (2.16 mg, orange solid) with a yield of 3%.
MS m/z(ESI)：492.1[M+1] + 。
HPLC:99.74％(214nm),98.80％(254nm)。
1 H NMR(400MHz,d 6- DMSO)δ11.82(s,1H),8.13-8.11(m,1H),7.93-7.92(m,1H),7.81(d,J＝7.6Hz,1H),7.55-7.41(m,5H),7.26(s,1H),6.99(d,J＝11.6Hz,1H),6.60(d,J＝7.6Hz,1H),4.71(d,J＝5.6Hz,2H),3.88(s,3H).
Example 43
Preparation of 3- [ 2-fluoro-5- (isoquinoline-8-amino) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-043)
Figure BDA0003608368340000931
First step of
Preparation of isoquinoline-8-carboxylic acid
Isoquinoline-8-carboxylic acid methyl ester 111a (300mg, 1.75mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL) followed by the addition of sodium hydroxide (240mg, 4.81mmol). The reaction was stirred at room temperature for 1 hour. Upon completion of LCMS detection reaction, organic solvent was distilled off under reduced pressure and water (20 mL) was added, followed by lyophilization to give isoquinoline-8-carboxylic acid 111b (250 mg, white solid) in yield: 81 percent.
MS m/z(ESI)：174.1[M+1] + 。
Second step of
Preparation of isoquinoline-8-carbonyl chloride
Preparation of isoquinoline-8-carboxylic acid 111b (300mg, 1.73mmol) was dissolved in dichloromethane (10 mL) followed by addition of DMF (0.1 mL) and oxalyl chloride (439mg, 3.46mmol). Stir at rt for 1h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and the solvent was removed to obtain the desired product, isoquinoline-8-carbonyl chloride 111c (300 mg, white solid), in yield: 81 percent.
MS m/z(ESI)：188.2(M-Cl+CH 3 )。
The third step
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-carboxamide
4-fluoro-2-methoxy-5-nitroaniline (582mg, 3.13mmol) was dissolved in ethyl acetate (20 mL), followed by addition of pyridine (247mg, 3.13mmol), and finally isoquinoline-8-carbonyl chloride 111c (300mg, 1.56mmol) was dissolved in ethyl acetate (5 mL) and added to the reaction solution under ice bath. The reaction was stirred at 0 ℃ for 2h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), drying over anhydrous sodium sulfate, and the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 55/45) to give N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-carboxamide 111d (50 mg, yellow solid), yield: 8 percent.
MS m/z(ESI)：342.1[M+1] + 。
The fourth step
Preparation of N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide
N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-carboxamide 111d (50mg, 0.15mmol) is dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (82mg, 1.46mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and then extracted with dichloromethane (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide 111e (40 mg, light yellow oil) as a yield: 79 percent.
MS m/z(ESI)：312.1[M+1] + 。
The fifth step
Preparation of 4- ({ [ 2-fluoro-5- (isoquinolin-8-amino) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide 111e (30mg, 0.10mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (39mg, 0.12mmol) and triethylamine (29mg, 0.29mmol). The reaction solution was stirred at 25 ℃ for 16 hours. Upon completion of the reaction monitored by LCMS, the compound dimethyl 4- ({ [ 2-fluoro-5- (isoquinolin-8-amino) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 111f (60 mg, light yellow liquid) was obtained in yield: 56 percent.
MS m/z(ESI)：553.1[M+1] + 。
The sixth step
Preparation of 3- [ 2-fluoro-5- (isoquinolin-8-amino) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction solution in the fifth step. Lithium hydroxide monohydrate (7mg, 0.16mmol) was added. The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give the crude product, which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- [ 2-fluoro-5- (isoquinoline-8-amino) -4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-043 (5 mg, light yellow solid), yield: 18 percent.
MS m/z(ESI)：507.1[M+1] + 。
HPLC:100％(214nm),99.67％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.57(s,1H),11.96(s,1H),10.09(s,1H),9.63(s,1H),8.58(d,J＝5.6Hz,1H),8.13(d,J＝7.6Hz,1H),7.99(d,J＝8.0Hz,1H),7.95-7.90(m,2H),7.90-7.84(m,1H),7.30(d,J＝11.6Hz,2H),3.94(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-122.09.
Example 44
Preparation of 3- (2-fluoro-4-methoxy-5- (phenylsulfonamido) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-044)
Figure BDA0003608368340000951
First step of
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) benzenesulfonamide
4-fluoro-2-methoxy-5-nitroaniline 112a (2000mg, 10.74mmol) was dissolved in a dichloromethane (30 mL) solution, pyridine (850mg, 10.7446mmol) was added, and the mixture was stirred at 0 ℃ for 5min. Benzenesulfonyl chloride (2087mg, 11.82mmol) was then added dropwise to the reaction mixture. The reaction was carried out at 25 ℃ for 3 hours under a nitrogen balloon atmosphere. After completion of the reaction, water (50 mL) was added to the reaction mixture. The reaction was extracted with dichloromethane (3X 30 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 2/1) to give N- (4-fluoro-2-methoxy-5-nitrophenyl) benzenesulfonamide 112b (2600 mg, yellow solid), yield: 66.74%.
MS m/z(ESI)：327.0[M+1] + 。
Second step of
Preparation of tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) (phenylsulfonyl) carbamate
N- (4-fluoro-2-methoxy-5-nitrophenyl) benzenesulfonamide 112b (200mg, 0.61mmol) was dissolved in a solution of dichloromethane (10 mL). Di-tert-butyl dicarbonate (267mg, 1.23mmol) and triethylamine (186mg, 1.84mmol) were added in this order. The reaction mixture was reacted at 25 ℃ for 12 hours. After completion of the reaction, water (20 mL) was added to the reaction mixture. Then extracted with ethyl acetate (3X 10 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and concentrated. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate = 3/1) to give tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) (benzenesulfonyl) carbamate 112c (190 mg, white solid) in yield: 65.43 percent.
MS m/z(ESI)：449.0(M+23)。
The third step
Preparation of N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (phenylsulfonyl) carbamic acid tert-butyl ester
Tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) (benzenesulfonyl) carbamate 112c (150mg, 0.35mmol) was dissolved in methanol (20 mL), and ammonium chloride (21mg, 0.39mmol) was added, followed by iron powder (196mg, 3.52mmol). The resulting mixture was stirred at 25 ℃ for 3 hours, after completion of the reaction. The resulting mixture was washed with 30mL of water. The resulting mixture was then extracted with ethyl acetate (3X 20 mL). The organic phases were collected, combined and dried over anhydrous sodium sulfate and concentrated. The resulting residue was further purified by silica gel column (petroleum ether/ethyl acetate = 2/1), N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (benzenesulfonyl) carbamic acid tert-butyl ester 112d (50 mg, white solid), yield: 32.26 percent.
MS m/z(ESI)：297.0(M+1-100)。
The fourth step
Preparation of 3- (5- (N- (tert-butoxycarbonyl) phenylsulfonamido) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Tert-butyl N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (phenylsulfonyl) carbamate 112d (50mg, 0.14mmol) and dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylate (71mg, 0.21mmol) and triethylamine (144mg, 1.42mmol) were dissolved in tetrahydrofuran (20 mL). The resulting mixture was stirred at 25 ℃ for 12h. Water (10 mL) and methanol (15 mL) were added, and the mixture was stirred at 25 ℃ for 1h, followed by addition of lithium hydroxide (34mg, 1.42mmol). The resulting mixture was stirred at 25 ℃ for 1h. After completion of the reaction, the resulting mixture was washed with 30mL of water. Then extracted with ethyl acetate (3X 20 mL) to give a mixture. The organic phases were collected, combined and dried over anhydrous sodium sulfate and concentrated. The obtained residue was further purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain 112e (60 mg, white solid) of 3- (5- (N- (tert-butoxycarbonyl) phenylsulfonamido) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (yield: 69.49%.
MS m/z(ESI)：492.0(M+1-100)。
The fifth step
Preparation of 3- (2-fluoro-4-methoxy-5- (phenylsulfonamido) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
112e (50mg, 0.09mmol) of 3- (5- (N- (tert-butoxycarbonyl) phenylsulfonamido) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (50mg, 0.09mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (104mg, 0.91mmol) was added. The resulting mixture was stirred at 25 ℃ for 30 minutes, and after completion of the reaction, concentrated. The concentrated solution was purified by column chromatography (mobile phase: acetonitrile/water = 3/1) to give a crude product, which was purified by preparative column (mobile phase: 0.1% formic acid/acetonitrile/water) to give 3- (2-fluoro-4-methoxy-5- (phenylsulfonamido) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-044 (21.9 mg, white solid) with yield: 46.88%.
MS m/z(ESI)：492.0[M+1] + 。
HPLC:98.48％(214nm),96.28％(254nm)。
1 H NMR(400MHz,d 6 -DMSO)δ14.55(s,1H),11.96(s,1H),9.64(s,1H),7.73-7.50(m,5H),7.43(d,J＝8.0Hz,1H),7.36(s,1H),7.05(d,J＝11.6Hz,1H),3.49(s,3H).
Example 45
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-045)
Figure BDA0003608368340000961
First step of
Preparation of N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (chloromethyl) -3,4-difluoro-1-methoxybenzene 113a (200mg, 1.0mmol) was dissolved in acetonitrile (5 mL) solution, followed by addition of 4-fluoro-2-methoxy-5-nitroaniline (386mg, 2.0mmol), potassium carbonate (717mg, 5.2mmol), and potassium iodide (34mg, 0.2mmol). The mixture was stirred at 50 ℃ for 2 hours under a nitrogen atmosphere. After completion of the reaction, the reaction solution was quenched with water and extracted with ethyl acetate (3X 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by chromatography column (ethyl acetate/petroleum ether = 0-20%) to give N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 113b (80 mg, yellow solid), yield: 20 percent.
MS m/z(ESI)：343.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.58(d,J＝7.4Hz,1H),7.07-7.01(m,1H),6.58(t,J＝8.9Hz,2H),4.47(s,2H),3.91(d,J＝5.8Hz,6H).
Second step of
Preparation of N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine
N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 113b (80mg, 0.23mmol) was dissolved in acetonitrile (5 mL) and iron powder (130mg, 2.3 mmol) and saturated ammonium chloride solution (5 mL) were added to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 113c (60 mg, yellow solid), yield: 70 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.1[M+1] + 。
The third step
Preparation of dimethyl 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 113c (60mg, 0.19mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (343mg, 1.0 mmol) and triethylamine (52mg, 0.51mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 113d (35 mg, yellow solid), yield: 33 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：554.1[M+1] + 。
The fourth step
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 113d (35mg, 0.06mmol) was dissolved in tetrahydrofuran/methanol/water =1:1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography (water/acetonitrile = 40%) to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-045 (15 mg, yellow solid) yield: 49 percent.
MS m/z(ESI)：508.0[M+1] + 。
HPLC:98.04％(214nm),96.30％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.67-14.60(m,1H),11.95(s,1H),7.42-7.27(m,2H),6.96(d,J＝11.4Hz,1H),6.88-6.82(m,1H),6.78(d,J＝7.6Hz,1H),4.83-4.80(m,1H),4.21(s,2H),3.82(d,J＝9.4Hz,6H).
Example 46
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-046)
Figure BDA0003608368340000971
First step of
Preparation of N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (chloromethyl) -3,4-difluoro-1-methoxybenzene 114a (200mg, 1.0mmol) was dissolved in acetonitrile (5 mL) solution, followed by addition of 4-fluoro-2-methoxy-5-nitroaniline (386 mg, 2.0mmol), potassium carbonate (717mg, 5.2mmol), and potassium iodide (34mg, 0.2mmol). The mixture was stirred at 50 ℃ for 2 hours under a nitrogen atmosphere. After completion of the reaction, the reaction solution was quenched with water and extracted with ethyl acetate (3X 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by chromatography (ethyl acetate/petroleum ether = 0-20%) to give N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 114b (80 mg, yellow solid), yield: 20 percent.
MS m/z(ESI)：343.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.58(d,J＝7.4Hz,1H),7.07-7.01(m,1H),6.58(t,J＝8.9Hz,2H),4.47(s,2H),3.91(d,J＝5.8Hz,6H).
Second step of
Preparation of N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline
N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 114b (80mg, 0.23mmol) was dissolved in methanol (5 mL), followed by addition of paraformaldehyde (36mg, 0.58mmol) and acetic acid (0.35mg, 0.0058mmol) to the solution. The reaction was stirred at room temperature for 0.5 h under nitrogen. Subsequently, sodium cyanoborohydride (18mg, 0.29mmol) was added to the system, and the reaction solution was stirred at room temperature for 2 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 114c (60 mg, yellow solid) in yield: 70 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：357.1[M+1] + 。
The third step
Preparation of N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxy-N1-toluene-1,3-diamine
N- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 114c (60mg, 0.19mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (130mg, 2.3mmol) and saturated ammonium chloride solution (5 mL) to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxy-N1-toluene-1,3-diamine 114d (50 mg, yellow solid) in yield: 85 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：327.1[M+1] + 。
The fourth step
Preparation of dimethyl 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
N1- (2,3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxy-N1-toluene-1,3-diamine 114d (50mg, 0.15mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (343mg, 1.0 mmol) and triethylamine (52mg, 0.51mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 114e (35 mg, yellow solid) yield: 33 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：568.1[M+1] + 。
The fifth step
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 114e (35mg, 0.06mmol) was dissolved in tetrahydrofuran/methanol/water =1:1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography (water/acetonitrile = 40%) to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid, cpd-046 (15 mg, yellow solid), yield: 49 percent.
MS m/z(ESI)：522.0[M+1] + 。
HPLC:98.81％(214nm),97.53％(254nm).
1 H NMR(400MHz,DMSO)δ14.65(s,1H),11.90(s,1H),7.36(s,1H),7.30-7.22(m,1H),7.07(d,J＝11.8Hz,1H),6.80-6.68(m,2H),4.41-4.32(m,5H),3.91(s,3H),3.50(s,3H).
Example 47
Preparation of 3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-047)
Figure BDA0003608368340000991
First step of
Preparation of 8- (benzylthio) isoquinoline
8-bromoisoquinoline (1.8g, 8.6 mmol), tris (dibenzylideneacetone) dipalladium (398mg, 0.4mmol) and 4,5-bis-diphenylphosphine-9,9-dimethylxanthene (501mg, 0.9mmol) were dissolved in 1,4 dioxane (20 mL), phenylmethanethiol (1.28g, 10.3mmol) and N, N-diisopropylethylamine (2.22g, 17.2 mmol) were added, and the reaction solution was stirred at 110 ℃ for 16h under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 9/1) to obtain 8- (benzylthio) isoquinoline 115b (2.1 g, yellow solid) in a yield of 95%.
MS m/z(ESI)：252.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.77(s,1H),8.56(d,J＝5.8Hz,1H),7.69-7.67(m,1H),7.62(d,J＝5.6Hz,1H),7.55-7.53(m,2H),7.26-7.24(m,5H),4.20(s,2H)。
Second step of
Preparation of isoquinoline-8-sulfonyl chloride
8- (benzylthio) isoquinoline 115b (1.56g, 6.2mmol) was dissolved in acetonitrile (20 mL) and acetic acid: water =2:1 (15 mL), 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (1.47g, 7.4 mmol) was added under an ice water bath, and the reaction solution was reacted at 25 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 7/3) to obtain isoquinoline-8-sulfonyl chloride 115c (506 mg, yellow solid) in a yield of 35%.
MS m/z(ESI)：227.9[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.28(s,1H),8.76(d,J＝6.6Hz,1H),8.60(d,J＝6.4Hz,1H),8.34(d,J＝8.4Hz,1H),8.28(d,J＝7.2Hz,1H),8.22-8.16(m,1H)。
The third step
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-sulfonamide
Isoquinoline-8-sulfonyl chloride 115c (500mg, 2.2 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (347mg, 4.9mmol) were dissolved in dichloromethane (15 mL), pyridine (347mg, 4.4 mmol) was added at room temperature, and the reaction mixture was reacted at 25 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by reverse phase column (58% acetonitrile/water) to give N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-sulfonamide 115d (136 mg, yellow solid) in 16% yield.
MS m/z(ESI)：378.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.60(s,1H),10.07(s,1H),8.70(d,J＝5.6Hz,1H),8.27(d,J＝8.4Hz,1H),8.05-7.97(m,2H),7.83-7.76(m,1H),7.06(d,J＝13.4Hz,2H),3.12(s,3H)。
The fourth step
Preparation of N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-sulfonamide
Dissolving N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-sulfonamide 115d (40mg, 0.11mmol), iron powder (17mg, 0.31mmol) and ammonium chloride (17mg, 0.32mmol) in ethanol: water =1:1 (5 mL), and the reaction mixture was reacted at 80 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched by addition of water (10 mL) and extracted with ethyl acetate (3X 10 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-sulfonamide 115e (35 mg, yellow solid) in 81% yield.
MS m/z(ESI)：348.1[M+1] + 。
The fifth step
Preparation of 4- (3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-sulfonamide 115e (25mg, 0.07mmol) and dimethyl-4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (29mg, 0.08mmol) tetrahydrofuran (3 mL) were added with triethylamine (22mg, 0.2mmol) at room temperature, the reaction solution was reacted at room temperature for 16 hours, after the reaction was completed, water (10 mL) was added to the reaction solution to quench, extraction was performed with ethyl acetate (3X 10 mL), all organic phases were combined, anhydrous sodium sulfate was dried and concentrated, and the obtained residue was purified with a reverse phase column (54% acetonitrile/water) to give 4- (3- (2-fluoro-5- (isoquinoline-8-sulfonamide) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 115f (22 mg, white solid) in 49% yield.
MS m/z(ESI)：589.0[M+1] + 。
The sixth step
Preparation of 3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 115f (20mg, 0.03mmol) and lithium hydroxide (3mg, 0.1mmol) were dissolved in tetrahydrofuran: methanol: water =1:1:1 (3 mL), and the reaction mixture was reacted at room temperature for 1 hour. After completion of the reaction, the reaction mixture was quenched by addition of 1M hydrochloric acid (5 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The residue obtained was purified by HPLC prep (mobile phase: acetonitrile/water) to give 3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-047 (0.72 mg, white solid) in 4% yield.
MS m/z(ESI)：543.0[M+1] + 。
HPLC:96.95％(214nm),91.19％(254nm)。
1 H NMR(400MHz,MeOD-d 4 )δ10.10(s,1H),8.58(s,1H),8.13(d,J＝8.0Hz,2H),7.96(s,1H),7.78(s,1H),7.52(s,1H),6.94(s,1H),6.90(d,J＝8.0Hz,1H),6.55(s,1H),3.01(s,3H)。
Example 48
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-048)
Figure BDA0003608368340001011
First step of
Preparation of methyl 1-methoxyisoquinoline-8-carboxylate
Methyl 1-chloroisoquinoline-8-carboxylate 116a (160mg, 0.72mmol) and sodium methoxide (195mg, 3.61mmol) were dissolved in methanol (2 mL). The mixture was reacted at 90 ℃ for 4 hours in a microwave apparatus. After the completion of the reaction, the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 3/1) to give methyl 1-methoxyisoquinoline-8-carboxylate 116b (110 mg, white solid) in 56% yield.
MS m/z(ESI)：218.1[M+1] + 。
Second step of
Preparation of (1-methoxyisoquinolin-8-yl) methanol
Methyl 1-methoxyisoquinoline-8-carboxylate 116b (100mg, 0.46mmol) was dissolved in tetrahydrofuran (5 mL) and DABAL-H/THF solution (2.5 mL) was slowly added dropwise and the mixture stirred at 0 ℃ for 1H. After the reaction was completed, a saturated ammonium chloride solution (5 mL) was added and quenched, and extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified with a silica gel column (petroleum ether/ethyl acetate = 2/1) to obtain (1-methoxyisoquinolin-8-yl) methanol 116c (90 mg, white solid) in yield: 82 percent.
MS m/z(ESI)：190.1[M+1] + 。
The third step
Preparation of 8- (chloromethyl) -1-methoxyisoquinoline
(1-Methoxyisoquinolin-8-yl) methanol 116c (60mg, 0.31mmol), tsCl (121mg, 0.63mmol), DMAP (8mg, 0.06mmol) and triethylamine (64mg, 0.63mmol) were dissolved in dichloromethane (5 mL). The mixture was stirred at 25 ℃ for 1 hour. After the completion of the reaction, the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 3/1) to give 8- (chloromethyl) -1-methoxyisoquinoline 116d (40 mg, white solid) in a yield of 53%.
MS m/z(ESI)：208.1[M+1] + 。
The fourth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1-methoxyisoquinoline
8- (chloromethyl) -1-methoxyisoquinoline 116d (40mg, 0.19mmol), 4-fluoro-2-methoxy-5-nitrophenol (43mg, 0.23mmol), potassium carbonate (53mg, 0.39mmol) and potassium iodide (48mg, 0.29mmol) were dissolved in acetonitrile (5 mL). The mixture was reacted at 80 ℃ for 2 hours. After completion of the reaction, the reaction mixture was extracted with dichloromethane (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1-methoxyisoquinoline 116e (50 mg, orange solid) in yield: and 64 percent.
MS m/z(ESI)：359.0[M+1] + 。
The fifth step
Preparation of 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] aniline
8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1-methoxyisoquinoline 116e (50mg, 0.14mmol) is dissolved in NH 4 To a Cl/MeCN (4 mL) solution, iron powder (156mg, 2.79mmol) was added, and the mixture was stirred at 25 ℃ for 1 hour. The reaction mixture was filtered and concentrated to give 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy]Aniline 116f (35 mg, brown solid) in 53% yield.
MS m/z(ESI)：329.1[M+1] + 。
The sixth step
Preparation of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] aniline 116f (35mg, 0.11mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (179mg, 0.53mmol) were dissolved in tetrahydrofuran (5 mL) then triethylamine (59mg, 1.06mmol) was added and the mixture stirred at 25 ℃ for 16 hours the reaction solution was not worked up to give a stock of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 116g (bright yellow solution) which was used directly in the next reaction.
MS m/z(ESI)：570.1[M+1] + 。
Seventh step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolving 116g (5 mL of stock solution) of dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate with lithium hydroxide (8mg, 0.35mmol) in methanol: water =3:1 (4 mL). The mixture was reacted at 25 ℃ for 1 hour. After the reaction was finished, the reaction mixture was directly concentrated and preliminarily purified by reverse column (water: acetonitrile = 3:2) to prepare 3- { 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-048 (2.46 mg, light yellow solid) with a yield of 6%.
MS m/z(ESI)：524.0[M+1] + 。
HPLC:99.38％(214nm),97.78％(254nm)。
1 H NMR(400MHz,d 6 -DMSO)δ11.95(s,1H),8.00(d,J＝5.8Hz,1H),7.87-7.85(m,1H),7.74(d,J＝5.2Hz,2H),7.42(d,J＝5.8Hz,1H),7.36(s,1H),7.24(d,J＝7.2Hz,1H),7.17(d,J＝11.6Hz,1H),5.63(s,2H),3.91(s,3H),3.85(s,3H).
Example 49
Preparation of 3- [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-049)
Figure BDA0003608368340001021
First step of
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
2-bromo-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 121d (180mg, 0.70mmol) and methylbenzeneboronic acid (78mg, 1.30mmol) were dissolved in toluene/ethanol/water (2ml, 7. Potassium carbonate (181mg, 1.31mmol) and ferrocene palladium dichloride (151mg, 0.13mmol) were then added and the reaction was reacted with a microwave at 80 ℃ for 1 hour under nitrogen. After completion of the reaction, the solvent was removed under reduced pressure, water (2 mL) was added, extraction was performed with ethyl acetate (3X 10 mL), and the mixture was washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 44- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 120b (50 mg, yellow solid) in 21% yield.
MS m/z(ESI)：335.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.08(s,1H),7.97(d,J＝8.0Hz,1H),7.84(d,J＝7.5Hz,1H),7.69(d,J＝7.2Hz,1H),7.49(t,J＝7.7Hz,1H),6.72(dd,J＝12.4,4.8Hz,1H),5.77(d,J＝5.6Hz,2H),3.96(s,4H).
Second step of
Preparation of 5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyaniline
4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 120b (50mg, 0.15mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (84mg, 1.50mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyaniline 120c (40 mg, brown solid) in 88% yield.
MS m/z(ESI)：305.2[M+1] + 。
The third step
Preparation of dimethyl 4- ({ [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyaniline 120c (47mg, 0.15mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (104mg, 0.31mmol) were dissolved in tetrahydrofuran (5 mL). Then, triethylamine (47mg, 0.46mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction solution is not subjected to any post-treatment to obtain a stock solution of 4- ({ [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dimethyl dicarboxylate 120d.
MS m/z(ESI)：546.0[M+1] + 。
The fourth step
Preparation of 3- [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 121d (5 mL stock solution) was dissolved in methanol with lithium hydroxide (13mg, 0.53mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for hours. After the reaction was finished, the reaction mixture was directly concentrated and subjected to preliminary purification by a reverse column (water: acetonitrile = 20) to prepare and purify 3- [5- (1,3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-049 (6.45 mg, white solid) with a yield of 8.80%.
MS m/z(ESI)：500.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,CDCl 3 )δ9.21(s,1H),8.04(d,J＝7.6Hz,1H),7.71(d,J＝6.8Hz,1H),7.51(t,J＝7.6Hz,1H),7.30(s,1H),7.15(d,J＝7.2Hz,1H),7.01(d,J＝11.2Hz,1H),5.62(s,2H),3.90(s,3H).
19 F NMR(376MHz,CDCl 3 )δ-129.22.
Example 50
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] phenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-050)
Figure BDA0003608368340001041
First step of
Preparation of 2-bromo-4-methyl-1,3-benzothiazole
4-methyl-1,3-benzothiazol-2-amine 121a (2.00g, 12.20mmol) was dissolved in acetonitrile (25 mL). Cuprous bromide (2.10g, 14.60mmol) and tert-butyl nitrite (1.89g, 0.02mmol) were added in this order. The reaction mixture was reacted at room temperature for 0.5 hour. After the reaction was completed, pH =6-7 was adjusted with a dilute hydrochloric acid solution, followed by extraction with ethyl acetate (3 × 10 mL) and washing with 1M hydrochloric acid (10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 99/1) to give 2-bromo-4-methyl-1,3-benzothiazole 121b (2.58 g, orange solid) with a yield of 88%.
MS m/z(ESI)：228.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.62(dd,J＝7.6,0.8Hz,1H),7.33-7.25(m,2H),2.71(s,3H).
Two steps of
Preparation of 2-bromo-4- (bromomethyl) -1,3-benzothiazole
2-bromo-4-methyl-1,3-benzothiazole 121b (500mg, 2.19mmol) was dissolved in carbon tetrachloride (10 mL) and benzoyl peroxide (106mg, 0.44mmol) and N-bromosuccinimide (468mg, 2.63mmol) were added at room temperature. The reaction mixture was reacted at 85 ℃ for 20 hours. After completion of the reaction, the reaction mixture was concentrated to remove carbon tetrachloride, diluted with dichloromethane (15 mL), washed with water (5 mL) and saturated brine (2X 5 mL) in this order, the organic phases were combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 80/20) to give 2-bromo-4- (bromomethyl) -1,3-benzothiazole 121c (670 mg, pale yellow solid) in 90% yield.
MS m/z(ESI)：308.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.76(dd,J＝8.0,1.2Hz,1H),7.56-7.50(m,1H),7.40(t,J＝7.8Hz,1H),4.99(s,2H).
The third step
Preparation of 2-bromo-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
2-bromo-4- (bromomethyl) -1,3-benzothiazole 121c (300mg, 0.98mmol) was dissolved in acetonitrile (10 mL), 4-fluoro-2-methoxy-5-nitrophenol (183mg, 0.98mmol), potassium carbonate (81mg, 0.49mmol) and potassium iodide (81mg, 0.49mmol) were added in this order, and the reaction solution was reacted at 65 ℃ for 1 hour. After completion of the reaction, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 2-bromo-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 121d (280 mg, yellow solid) in 62% yield.
MS m/z(ESI)：413[M+1] + 。
The fourth step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzothiazole
2-bromo-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 121d (180mg, 0.70mmol) and methylbenzeneboronic acid (78mg, 1.30mmol) were dissolved in toluene/ethanol/water (2ml, 7. Potassium carbonate (181mg, 1.31mmol) and ferrocene palladium dichloride (151mg, 0.13mmol) were then added and the reaction was reacted in a microwave at 80 ℃ for 1 hour under nitrogen. After completion of the reaction, the solvent was removed under reduced pressure, water (2 mL) was added, extraction was performed with ethyl acetate (3X 10 mL), and the mixture was washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzothiazole 121e (40 mg, yellow solid) in 24% yield.
MS m/z(ESI)：349.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.95(d,J＝7.3Hz,1H),7.82(d,J＝8.0Hz,1H),7.63(d,J＝7.5Hz,1H),7.39(t,J＝7.7Hz,1H),6.72(d,J＝12.4Hz,1H),5.75(s,2H),3.97(s,3H),2.93(s,3H).
The fifth step
Preparation of 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] aniline
4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzothiazole 121e (40mg, 0.11mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (64mg, 1.15mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] aniline 121f (40 mg, brown solid) in 88% yield.
MS m/z(ESI)：319.2[M+1] + 。
The sixth step
4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
Preparation of esters
2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] aniline 121f (35mg, 0.11mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (74mg, 0.22mmol) were dissolved in tetrahydrofuran (5 mL). Then, triethylamine (33mg, 0.33mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction mixture was not subjected to any post-treatment to give 121g of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate as a raw material.
MS m/z(ESI)：560.2[M+1] + 。
Seventh step
4- [ ({ 5- [ (5,6-difluoro-2,3-dihydro-1,4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -3- (methoxycarbonyl)
Preparation of thiophene-2-carboxylic acid
Dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 121g (5 mL stock solution) was dissolved in methanol with lithium hydroxide (13mg, 0.53mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and subjected to preliminary purification by a reverse column (water: acetonitrile = 20) to obtain 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzothiazol-4-yl) methoxy ] phenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-050 (4.30 mg, white solid) with a yield of 9.28%.
MS m/z(ESI)：514.1[M+1] + 。
HPLC:99.47％(214nm),100％(254nm).
1 H NMR(400MHz,CDCl 3 )δ7.89(d,J＝7.2Hz,1H),7.64(d,J＝7.6Hz,1H),7.41(t,J＝7.6Hz,1H),7.31(s,1H),7.16(d,J＝7.2Hz,1H),7.01(d,J＝11.2Hz,1H),5.55(s,2H),3.91(s,3H),2.80(s,3H).
19 F NMR(376MHz,CDCl 3 )δ-129.22.
Example 51
Preparation of 3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-051)
Figure BDA0003608368340001061
First step of
Preparation of 1H benzo [ d ] [1,2,3] triazole-4-carboxylic acid
2,3-diaminobenzoic acid (1000mg, 6.6mmol) 122a was dissolved in acetic acid (6 mL) and water (9 mL). A solution of sodium nitrite (544mg, 7.9 mmol) in water (5 mL) was then gradually added dropwise to the mixture, and the reaction mixture was stirred at room temperature for 18 hours. After completion of the reaction, filtration and washing of the solid with water gave 1H-1,2,3-benzotriazole-4-carboxylic acid 122b (1100 mg, grey solid) in yield: 92 percent.
MS m/z(ESI)：164.1[M+1] + 。
1 H NMR(400MHz,DMSO-d6)δ8.36(d,J＝8.2Hz,1H),8.12(d,J＝7.2Hz,1H),7.60-7.38(m,1H).
Second step of
Preparation of 1H-1,2,3-benzotriazole-4-carboxylic acid methyl ester
1H-1,2,3-benzotriazole-4-carboxylic acid 122b (1100mg, 6.6 mmol) was dissolved in methanol (20 mL), and then thionyl chloride (2188mg, 18mmol) was gradually added dropwise to the solution at 0 ℃ and the reaction solution was stirred at 40 ℃ for 16 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on silica gel column (MeOH/DCM = 10%) to give 1H-1,2,3-benzotriazole-4-carboxylic acid methyl ester 122c (906 mg, grey solid), yield: 68 percent.
MS m/z(ESI)：178.1[M+1] + 。
The third step
Preparation of 1H-1,2,3-benzotriazol-4-yl methanol
Methyl 1H-1,2,3-benzotriazole-4-carboxylate 122c (900mg, 5.07mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and then a diisobutylaluminum hydride solution (1.0M in n-hexane) (10 mL) was added dropwise to the solution at 0 ℃. The mixture was stirred at 0 ℃ for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified on silica gel column (MeOH/DCM = 10%) to give 1H-1,2,3-benzotriazol-4-yl methanol 122d (540 mg, grey solid), yield: 60 percent.
MS m/z(ESI)：150.1[M+1] + 。
The fourth step
Preparation of 4- (chloromethyl) -1H benzo [ d ] [1,2,3] triazole
1H-1,2,3-benzotriazol-4-ylmethanol 122d (540mg, 3.60mmol) was dissolved in ultra dry tetrahydrofuran (6 mL), followed by addition of thionyl chloride (10 mL) at 0 ℃ under nitrogen protection. The reaction was stirred at room temperature for 3 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 122e of 4- (chloromethyl) -1H benzo [ d ] [1,2,3] triazole (363 mg, yellow solid), yield: 60 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：168.1[M+1] + 。
The fifth step
Preparation of 4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1H-benzo [ d ] [1,2,3] triazole
4- (chloromethyl) -1H-benzo [ d ] [1,2,3] triazole 122e (363mg, 2.16mmol)), 4-fluoro-2-methoxy-5-nitroaniline (522mg, 2.80mmol), potassium carbonate (2013mg, 10.8mmol), and potassium iodide (52mg, 0.21mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50 ℃ for two hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on a silica gel column (MeOH/DCM = 10%) to give 4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1H-benzo [ d ] [1,2,3] triazole 122f (300 mg, yellow solid) yield: 44 percent.
MS m/z(ESI)：319.1[M+1] + 。
The sixth step
Preparation of 5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline
4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1H-benzo [ d ] [1,2,3] triazole 122f (300mg, 0.94mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (650mg, 9.4mmol) and saturated ammonium chloride solution (5 mL) to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 122g (200 mg, yellow solid) of 5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline, yield: 74 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：289.1[M+1] + 。
Seventh step
Preparation of 4- (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2,3-dicarboxylic acid dimethyl ester
122g (200mg, 0.64mmol) of 5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline was dissolved in tetrahydrofuran (6 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (686mg, 2.0 mmol) and triethylamine (162mg, 1.52mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 4- (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2,3-dicarboxylic acid dimethyl ester 122H (85 mg, yellow solid) yield: 27 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：530.1[M+1] + 。
Eighth step
Preparation of 3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolving (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2,3-dicarboxylic acid dimethyl ester 122H (85mg, 0.16mmol) in tetrahydrofuran/methanol/water =1:1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative purification (FA) to give 3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-051 (8 mg, yellow solid) in yield: 10 percent.
MS m/z(ESI)：484.1[M+1] + 。
HPLC:95.98％(214nm),89.42％(254nm).
1 H NMR(400MHz,DMSO-d6)δ15.99-15.84(m,1H),14.66-14.42(m,1H),11.97(s,1H),7.65-7.27(m,5H),7.16(d,J＝11.6Hz,1H),5.39(s,2H),3.82(s,3H).
Example 52
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzooxazol-4-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-052)
Figure BDA0003608368340001081
First step of
Preparation of 2-methyl-1,3-benzoxazole-4-carboxylic acid
123a (2.00g, 0.01mol) 2-amino-3-hydroxybenzoic acid was dissolved in trimethyl orthoacetate (20 mL) and stirred at 90 ℃ for 3 hours. After the reaction was completed, water (10 mL) was added to quench, extraction was performed with ethyl acetate (3 × 50 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to obtain preparation 123b of 2-methyl-1,3-benzoxazole-4-carboxylic acid (1.45 g, orange solid), yield: and 63 percent.
MS m/z(ESI)：178.2[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ13.03(s,1H),7.91(dd,J＝8.0,1.2Hz,1H),7.85(dd,J＝7.8,1.0Hz,1H),7.46-7.41(m,1H),2.67(s,3H).
Second step of
Preparation of (2-methyl-1,3-benzoxazol-4-yl) methanol
Preparation 123b (1.00g, 5.60mmol) of 2-methyl-1,3-benzoxazole-4-carboxylic acid was dissolved in tetrahydrofuran (5 mL), and 1N borane in tetrahydrofuran (5 mL) was added dropwise and reacted at 0 ℃ for 3 hours. After the reaction was completed, methanol was added dropwise to quench, and the reaction solution was extracted with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give (2-methyl-1,3-benzoxazol-4-yl) methanol 123c (356 mg, yellow solid) in yield: 39 percent.
MS m/z(ESI)：164.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.42–7.38(m,1H),7.27–7.24(m,2H),5.03(s,2H),2.62(s,3H).
The third step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzoxazole
Diisopropyl azodicarboxylate (289mg, 1.10mmol) was added dropwise to a solution of (2-methyl-1,3-benzoxazol-4-yl) methanol 123c (150mg, 0.92mmol), 4-fluoro-2-methoxy-5-nitrophenol (206mg, 1.10mmol) and triphenylphosphine (223mg, 1.10mmol) in tetrahydrofuran (10 mL) at 0 ℃ and stirred at 0 ℃ for 3 hours. After the reaction was completed, water quenching was added, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 4/1) to obtain 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzoxazole 123d (295 mg, white solid) in yield: 96 percent.
MS m/z(ESI)：333.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.87(d,J＝7.4Hz,1H),7.46(dd,J＝7.8,2.2Hz,2H),7.32(t,J＝7.8Hz,1H),6.72(d,J＝12.4Hz,1H),5.54(s,2H),3.95(s,3H),2.70(s,3H).
The fourth step
Preparation of 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzoxazol-4-yl) methoxy ] aniline
To a solution of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1,3-benzoxazole 123d (285mg, 0.86mmol) in ethanol (3 mL) and water (3 mL) were added iron powder (144mg, 2.57mmol) and ammonium chloride (138mg, 2.57mmol), and the mixture was reacted at 80 ℃ for 1 hour. After the reaction was completed, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to obtain 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzoxazol-4-yl) methoxy ] aniline 123e (246 mg, yellow solid), yield: 94 percent.
MS m/z(ESI)：303.1[M+1] + 。
The fifth step
Preparation of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzooxazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
To a solution of 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzoxazol-4-yl) methoxy ] aniline 123e (216mg, 0.71mmol) and triethylamine (217mg, 2.14mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (264mg, 0.79mmol) and reacted at 25 ℃ for 2 hours after completion of the reaction, water (5 mL) was added and quenched, extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to give dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-3425 zf-dicarboxylate (white solid yield: 40 mg, 123 mg).
MS m/z(ESI)：544.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.95(s,1H),8.82(s,1H),7.94(s,1H),7.81(d,J＝8.0Hz,1H),7.64(d,J＝8.0Hz,1H),7.45(d,J＝7.4Hz,1H),7.37(t,J＝7.8Hz,1H),7.02(d,J＝12.6Hz,1H),5.30(s,2H),3.89(s,3H),3.83(s,3H),3.75(s,3H),2.62(s,3H).
The sixth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzooxazol-4-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of dimethyl 4- [ ({ { 2-fluoro-4-methoxy-5- [ (2-methyl-1,3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 123f (134mg, 0.25mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide monohydrate (31mg, 0.74mmol), and the mixture was reacted at 25 ℃ for 1 hour. After the reaction was completed, 1M diluted hydrochloric acid was added dropwise to adjust pH =6-7, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give preparation Cpd-052 (5 mg, white solid) of 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid: 4 percent.
MS m/z(ESI)：498.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.66(d,J＝7.8Hz,1H),7.47(d,J＝7.4Hz,1H),7.37(t,J＝7.8Hz,1H),7.31(d,J＝7.4Hz,1H),7.11(d,J＝11.6Hz,1H),7.04(s,1H),5.26(s,2H),3.81(s,3H),2.61(s,3H).
Example 53
Preparation of 3- [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxa-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-053)
Figure BDA0003608368340001101
First step of
Preparation of 4- (bromomethyl) -1,3-benzodioxoles
1,3-benzodioxazol-4-ylmethanol 124a (250mg, 1.64mmol), triphenylphosphine (862mg, 3.29mmol) and carbon tetrabromide (1.00g, 3.29mmol) were dissolved in dichloromethane (10 mL). The mixture was stirred at 25 ℃ for 2 hours and then concentrated, and the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 4- (bromomethyl) -1,3-benzodioxole 124b (220 mg, white oil), yield: 50 percent.
1 H NMR(400MHz,CDCl 3 )δ6.82-6.78(m,3H),6.02(s,2H),4.47(s,2H).
Second step of
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzodioxole
4- (bromomethyl) -1,3-benzodioxole 124b (120mg, 0.56mmol), 4-fluoro-2-methoxy-5-nitrophenol (104mg, 0.56mmol), potassium carbonate (154mg, 1.11mmol) and potassium iodide (139mg, 0.84mmol) were dissolved in acetonitrile (5 mL), the mixture was stirred at 90 ℃ for 1 hour and concentrated, and the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 1/2) to give 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzodioxole 124c (80 mg, white solid) in yield: 35 percent.
MS m/z(ESI)：344.0(M+23)。
1 H NMR(400MHz,CDCl 3 )δ7.78(d,J＝7.2Hz,1H),6.94(dd,J＝7.6,1.6Hz,1H),6.86-6.79(m,2H),6.71(d,J＝12.4Hz,1H),6.04(s,2H),5.16(s,2H),3.95(s,3H).
The third step
Preparation of 5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyaniline
4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzodioxole 124c (80mg, 0.25mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL), iron powder (279mg, 4.98mmol) was added and the mixture was stirred at 25 ℃ for 1 hour. The reaction was filtered and concentrated to give preparation 124d of 5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyaniline (50 mg, brown oil). The yield was 55%.
MS m/z(ESI)：292.1[M+1] + 。
The fourth step
Preparation of dimethyl 4- ({ [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
Preparation of 5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyaniline 124d (50mg, 0.17mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (288mg, 0.86mmol) were dissolved in tetrahydrofuran (5 mL) then triethylamine (90mg, 1.72mmol) was added and the mixture was stirred at 25 ℃ for 16 hours the reaction solution was not subjected to any post-treatment to give a stock of dimethyl 4- ({ [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 124e (brilliant yellow solution) which was used directly in the next reaction.
MS m/z(ESI)：533.0[M+1] + 。
The fifth step
Preparation of 3- [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxa-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolve 4- ({ [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 124e (5 mL stock solution) with lithium hydroxide (27mg, 1.13mmol) in methanol: water =3:1 (4 mL). The mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and preliminarily purified by reverse column (water: acetonitrile = 3:2) to obtain 3- [5- (1,3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxa-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-053 (2.63 mg, yellow solid) with a yield of 4%.
MS m/z(ESI)：487.0[M+1] + 。
HPLC:98.36％(214nm),100％(254nm)。
1 H NMR(400MHz,d 6 -DMSO)δ11.99(s,1H),7.37(s,1H),7.26(d,J＝7.2Hz,1H),7.13(d,J＝11.6Hz,1H),6.94(m,2H),6.86(t,J＝7.6Hz,1H),6.02(s,2H),4.92(s,2H),3.83(s,3H).
Example 54
Preparation of 3- [ 2-fluoro-5- (1H-indazol-7-ylmethoxy) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-054)
Figure BDA0003608368340001111
First step of
Preparation of 1H-indazol-7-ylmethanol
Methyl 1H-indazole-7-carboxylate 125a (50mg, 0.28mmol) was dissolved in tetrahydrofuran (5 mL). Diisobutylaluminum hydride (5 mL) was then added at 0 ℃. The reaction mixture was reacted at 25 ℃ for 1 hour. Then, the mixture was quenched with a saturated ammonium chloride solution, extracted with dichloromethane (2X 10 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 50/50) to give 1H-indazol-7-ylcarbinol 125b (190 mg, white solid) in 81% yield.
MS m/z(ESI)：149.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ13.01(s,1H),8.06(d,J＝1.2Hz,1H),7.64(d,J＝8.0Hz,1H),7.31(dd,J＝7.0,0.8Hz,1H),7.08(dd,J＝8.0,7.12Hz,1H),5.27(t,J＝5.6Hz,1H),4.81(d,J＝5.6Hz,2H).
Second step of
Preparation of 7- (bromomethyl) -1H-indazole
1H-indazol-7-ylcarbinol 125b (120mg, 0.81mmol) was dissolved in dichloromethane (5 mL). Triphenylphosphine (424mg, 1.62mmol) and carbon tetrabromide (537mg, 1.62mmol) were then added. The reaction mixture was reacted at 25 ℃ for 1 hour. Then, dichloromethane (2X 5 mL) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 7- (bromomethyl) -1H-indazole 125c (100 mg, red solid) with a yield of 52%.
MS m/z(ESI)：211.0[M+1] + 。
The third step
Preparation of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1H-indazole
7- (bromomethyl) -1H-indazole 125c (100mg, 0.47mmol) and 4-fluoro-2-methoxy-5-nitrophenol (97mg, 0.52mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (196mg, 0.28mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 50/50) to give 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1H-indazole 125d (143 mg, white solid) with a yield of 85%.
MS m/z(ESI)：318.1[M+1] + 。
The fourth step
Preparation of 2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyaniline
7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1H-indazole 125d (140mg, 0.57mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (2 g) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyaniline 125e (112 mg, white solid) in 74% yield.
MS m/z(ESI)：288.1[M+1] + 。
The fifth step
Preparation of 4- ({ [ [ 2-fluoro-5- (1H-indazol-7-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyaniline 125e (30mg, 0.10mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (42mg, 0.12mmol) are dissolved in tetrahydrofuran (5 mL), then triethylamine (31mg, 0.31mmol) is added, the reaction solution reacts for 1 hour at 25 ℃, and the reaction solution is not subjected to any post-treatment to obtain a stock solution 4- ({ [ [ 2-fluoro-5- (1H-indazol-7-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 125f.
MS m/z(ESI)：529.1[M+1] + 。
The sixth step
Preparation of 3- [ 2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [ [ 2-fluoro-5- (1H-indazol-7-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 125f (5 mL stock solution) was dissolved in methanol with lithium hydroxide (3mg, 0.078mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, 3- [ 2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-054 (4.23 mg, white solid) was obtained after direct concentration, preliminary purification by reverse column (water: acetonitrile = 82).
MS m/z(ESI)：483.1[M+1] + 。
HPLC:98.73％(214nm),95.05％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ13.25(s,1H),8.10(s,1H),7.76(d,J＝8.0Hz,1H),7.40(d,J＝6.8Hz,1H),7.29(d,J＝7.2Hz,1H),7.23(s,1H),7.14-7.08(m,2H),5.26(s,2H),3.78(s,3H).
Example 55
Preparation of 3- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-055)
Figure BDA0003608368340001131
First step of
Preparation of 4-fluoro-2-methoxy-N-methyl-N- ((2-methyl-1H-isoquinolin-8-yl) methyl) -5-nitroaniline
4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline 126a (40mg, 0.12mmol) was dissolved in jiachun (10 mL), followed by addition of acetic acid (0.05 mL), paraformaldehyde (18mg, 0.61mmol), stirring of the mixture at room temperature for 30 min, and finally addition of sodium cyanoborohydride (23mg, 0.37mmol). The reaction solution was stirred at 55 ℃ for 48h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), and after drying over anhydrous sodium sulfate, the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 1/5) to give 4-fluoro-2-methoxy-N-methyl-N- ((2-methyl-1H-isoquinolin-8-yl) methyl) -5-nitroaniline 126b (20 mg, yellow solid), yield: 41 percent.
MS m/z(ESI)：360.2[M+1] + 。
Second step of
Preparation of 4-fluoro-6-methoxy-N1-methyl-N1- ((2-methyl-1H-isoquinolin-8-yl) methyl) benzene-1,3-diamine
4-fluoro-2-methoxy-N-methyl-N- ((2-methyl-1H-isoquinolin-8-yl) methyl) -5-nitroaniline 126b (40mg, 0.11mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by addition of iron powder (62mg, 1.11mmol). The reaction was stirred at 25 ℃ for 2h. After completion of the reaction the iron powder was removed by filtration and then extracted with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 4-fluoro-6-methoxy-N1-methyl-N1- ((2-methyl-1H-isoquinolin-8-yl) methyl) benzene-1,3-diamine 126c (30 mg, light yellow oil), yield: 65 percent.
MS m/z(ESI)：330.1[M+1] + 。
The third step
Preparation of dimethyl 4- (2- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) acetamido) thiophene-2,3-dicarboxylate
4-fluoro-6-methoxy-N1-methyl-N1- ((2-methyl-1H-isoquinolin-8-yl) methyl) benzene-1,3-diamine 126c (25mg, 0.08mmol) was dissolved in tetrahydrofuran (10 mL) followed by the addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (31mg, 0.09mmol) and triethylamine (23mg, 0.23mmol). The reaction solution was stirred at 25 ℃ for 16 hours. Upon completion of the reaction monitored by LCMS, the compound 4- (2- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) acetamido) thiophene-2,3-dicarboxylic acid dimethyl ester 126d (50 mg, light yellow liquid) was obtained, yield: 58 percent.
MS m/z(ESI)：571.2[M+1] + 。
The fourth step
3- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-
Preparation of formic acid
Methanol (2 mL) and water (1 mL) were added to the reaction solution in the fifth step. Lithium hydroxide monohydrate (7mg, 0.16mmol) was added. The reaction was stirred at room temperature for 1h. After the reaction was complete the solvent was evaporated to give the crude product, which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-055 (5 mg, pale yellow solid), yield: 18 percent.
MS m/z(ESI)：525.1[M+1] + 。
HPLC:98.26％(214nm),97.38％(254nm).
1 H NMR(400MHz,CD 3 OD)δ7.29-7.22(m 2H),7.16(d,J＝7.2Hz,1H),6.99-6.92(m,2H),6.79(d,J＝2.4Hz,1H),4.61-4.57(m,1H),4.41-4.37(m,1H),4.23-1.15(m,2H),3.93(s,3H),3.53(d,J＝6.4Hz,2H),3.20(d,J＝6.4Hz,2H),2.94(s,3H),2.57(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-126.02.
Example 56
Preparation of 3- { 2-fluoro-5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-056)
Figure BDA0003608368340001141
First step of
Preparation of [ (2-bromo-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine
2-bromo-3-fluorobenzaldehyde 127a (4.5g, 22.2mmol) was dissolved in ethanol (60 mL). 2,2-dimethoxyethylamine (2.57g, 24.4 mmol) was then added at 25 ℃. The reaction mixture was reacted at 80 ℃ for 16 hours. The solvent was then spun dry and extracted with ethyl acetate (2X 50 mL), the organic phases combined and dried over anhydrous sodium sulfate and concentrated to give [ (2-bromo-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 127b (4.23 g, yellow liquid) in 55% yield.
MS m/z(ESI)：292.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.29-7.22(m,2H),7.22-7.17(m,2H),7.03(td,J＝8.4,1.6Hz,2H),4.50(t,J＝5.6Hz,2H).
Second step of
Preparation of N- [ (2-bromo-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine
[ (2-bromo-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine 127b (4.2g, 1.86mmol) was dissolved in dichloromethane (60 mL). Triethylamine (4.36g, 0.04mmol) and 4-toluenesulfonyl chloride (8.24g, 0.04mmol) were then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then, it was extracted with water (2X 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give N- [ (2-bromo-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine 127c (4.2 g, red solid) in 58% yield.
MS m/z(ESI)：446.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.72(d,J＝8.4Hz,2H),7.38-7.27(m,4H),7.02(s,1H),5.30(s,1H),4.55(s,2H),3.30(d,J＝5.2Hz,2H),3.21(s,6H),2.44(s,3H).
The third step
Preparation of 8-bromo-7-fluoroisoquinoline
N- [ (2-bromo-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine 127c (3.6g, 0.008mmol) was dissolved in dichloromethane (10 mL) and added dropwise to a solution of anhydrous aluminum trichloride (6.4g, 0.048mmol) in dichloromethane (5 mL) at 0 ℃. The reaction mixture was reacted at 25 ℃ for 16 hours. Then, the mixture was quenched by addition of ice water and extracted with water (2X 50 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give 8-bromo-7-fluoroisoquinoline 127d (1.2 g, yellow solid) in 60% yield.
MS m/z(ESI)：226.0[M+1] + 。
The fourth step
Preparation of 7-fluoroisoquinoline-8-carboxylic acid methyl ester
8-bromo-7-fluoroisoquinoline 127d (1.2g, 5mmol) was dissolved in methanol (12 mL). Triethylamine (1.55g, 15mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium) (370mg, 0.0mmol) were then added thereto, and the mixture was sealed under pressure in a closed vessel filled with carbon monoxide, and the reaction mixture was reacted at 80 ℃ for 16 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 55/45) to give methyl 7-fluoroisoquinoline-8-carboxylate 127e (832 mg, brown solid) in 70% yield.
MS m/z(ESI)：206.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ 1 H NMR(400MHz,CDCl 3 )δ9.61(s,1H),8.61(d,J＝5.6Hz,1H),7.98(dd,J＝9.2,5.2Hz,1H),7.73(d,J＝5.6Hz,1H),7.55(t,J＝9.2Hz,1H),4.10(s,3H).
The fifth step
Preparation of (7-fluoroisoquinolin-8-yl) methanol
Methyl 7-fluoroisoquinoline-8-carboxylate 127e (200mg, 0.97mmol) was dissolved in tetrahydrofuran (5 mL). Then, lithium aluminum hydride (184mg, 4.87mmol) was added thereto at 0 ℃ and the reaction mixture was reacted at 0 ℃ for 0.5 hour. Then, the mixture was quenched by addition of ice water and extracted with dichloromethane (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give (7-fluoroisoquinolin-8-yl) methanol 127f (120 mg, yellow solid) in 66% yield.
MS m/z(ESI)：178.0[M+1] + 。
The sixth step
Preparation of 8- (bromomethyl) -7-fluoroisoquinoline
(7-Fluoroisoquinolin-8-yl) methanol 127f (70mg, 0.35mmol) and triethylamine (205mg, 2.03mmol) were dissolved in dichloromethane (5 mL). Then, p-toluenesulfonyl chloride (386mg, 2.03mmol) and 4-dimethylaminopyridine (8mg, 0.068mmol) were added to the reaction mixture, and the reaction mixture was reacted at 25 ℃ for 1 hour. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 127g (100 mg, brown solid) of 8- (bromomethyl) -7-fluoroisoquinoline in 55% yield.
MS m/z(ESI)：196.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ 1 H NMR(400MHz,CDCl 3 )δ9.62(s,1H),8.64(d,J＝5.6Hz,1H),7.91(dd,J＝9.2,5.2Hz,1H),7.77(d,J＝5.6Hz,1H),7.55(t,J＝9.2Hz,1H),5.17(d,J＝1.4Hz,2H).
Seventh step
Preparation of 7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
127g (100mg, 0.51mmol) of 8- (bromomethyl) -7-fluoroisoquinoline and 4-fluoro-2-methoxy-5-nitrophenol (105mg, 0.56mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (211mg, 1.5 mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 2 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 127h (62 mg, yellow solid) in 35% yield.
MS m/z(ESI)：347.0[M+1] + 。
Eighth step
Preparation of 5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 127h (60mg, 0.17mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (2 g) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 127i (50 mg, brown solid) in 55% yield.
MS m/z(ESI)：317.0[M+1] + 。
The ninth step
Preparation of 4- [ ({ [5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
5- [ (7-Fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 127i (30mg, 0.09mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (31mg, 0.09mmol) were dissolved in tetrahydrofuran (5 mL), then triethylamine (28mg, 0.28mmol) was added, and the reaction solution was reacted at 25 ℃ for 1 hour to obtain a stock solution 4- [ ({ [5- [ (7-Fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 127j without any post-treatment.
MS m/z(ESI)：558.1[M+1] + 。
The tenth step
Preparation of 3- {5- [ (7-Fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ [5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 127j (5 mL stock solution) was dissolved in methanol with lithium hydroxide (3mg, 0.078mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and subjected to preliminary purification by reverse column (water: acetonitrile = 82) to prepare and purify 3- {5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-056 (7.2 mg, white solid) with a yield of 50%.
MS m/z(ESI)：512.0[M+1] + 。
HPLC:100％(214nm),99.38％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ12.01(s,1H),9.59(s,1H),8.59(d,J＝5.6Hz,1H),8.16(dd,J＝9.2,5.2Hz,1H),7.94(d,J＝5.6Hz,1H),7.76(t,J＝9.2Hz,1H),7.43-7.37(m,2H),7.17(d,J＝11.6Hz,1H),5.55(d,J＝1.2Hz,2H),3.79(s,3H).
Example 57
Preparation of 3- (5- ((6,7-Difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-057)
Figure BDA0003608368340001161
First step of
Preparation of (2-bromo-3,4-difluorophenyl) methanol
2-bromo-3,4-difluorobenzoic acid 129a (5000mg, 21.1mmol) was dissolved in ultra-dry tetrahydrofuran (50 mL), followed by addition of borane tetrahydrofuran solution (1M) (10 mL) dropwise at 0 ℃ under a nitrogen atmosphere. After the reaction was completed, water was added to quench, and then the reaction solution was extracted with ethyl acetate (3 × 40 mL), and the combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by silica gel column (ethyl acetate/petroleum ether = 20%) to give (2-bromo-3,4-difluorophenyl) methanol 129b (4600 mg, white solid) in yield: 88 percent.
1 H NMR(400MHz,CDCl 3 )δ7.29-7.25(m,1H),7.19-7.13(m,1H),4.74(s,2H),1.96(s,1H).
Second step of
Preparation of 2-bromo-3,4-difluorobenzaldehyde
(2-bromo-3,4-difluorophenyl) methanol 129b (4600mg, 20.7mmol) was dissolved in tetrahydrofuran (20 mL), and manganese dioxide (3898mg, 44.9mmol) was gradually added to the solution, and the reaction solution was stirred at room temperature for 4 hours. After the reaction was complete, the solid residue was filtered off, the filtrate was extracted with ethyl acetate (3 × 20 mL), the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 2-bromo-3,4-difluorobenzaldehyde 129c (4000 mg, grey solid) yield: 86 percent.
1 H NMR(400MHz,CDCl 3 )δ10.28(d,J＝0.6Hz,1H),7.79-7.75(m,1H),7.231-7.25(m,1H).
The third step
Preparation of N- (2-bromo-3,4-difluorobenzyl) -2,2-dimethoxyethane-1-amine
2-bromo-3,4-difluorobenzaldehyde 129c (4000mg, 18.1mmol) was dissolved in ethanol solution (30 mL) followed by the addition of 2,2-dimethoxyethane-1-amine (2093mg, 19.9mmol). The reaction solution was stirred at 80 ℃ for 16 hours under a nitrogen atmosphere. After the reaction was completed, after the reaction solution was cooled to room temperature, sodium cyanoborocyanide (9099mg, 144.80mmol) was added, the reaction solution was stirred at room temperature for 1 hour, then the mixture was concentrated and extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (ethyl acetate/petroleum ether = 20%) to give N- (2-bromo-3,4-difluorobenzyl) -2,2-dimethoxyethane-1-amine 129d (3000 mg, yellow oil), yield: 48 percent.
MS m/z(ESI)：310.0[M+1] + 。
The fourth step
Preparation of N- (2-bromo-3,4-difluorobenzyl) -N- (2,2-dimethoxyethyl) -4-toluenesulfonamide
N- (2-bromo-3,4-difluorobenzyl) -2,2-dimethoxyethane-1-amine 129d (3000mg, 9.7mmol) was dissolved in e-dichloromethane solution (20 mL), followed by addition of triethylamine (4078mg, 40mmol) and 4-toluenesulfonyl chloride (4610mg, 24mmol) at 0 ℃. The reaction solution was stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified on a silica gel column (ethyl acetate/petroleum ether = 20%) to give N- (2-bromo-3,4-difluorobenzyl) -N- (2,2-dimethoxyethyl) -4-toluenesulfonamide 129e (3500 mg, white solid), yield: 79 percent.
MS m/z(ESI)：486.1(M+23)。
1 H NMR(400MHz,CDCl 3 )δ7.71(d,J＝8.2Hz,2H),7.36-7.27(m,3H),7.16-7.10(m,1H),4.48(s,2H),3.32-3.18(m,8H),2.45(s,3H).
The fifth step
Preparation of 8-bromo-6,7-difluoroisoquinoline
N- (2-bromo-3,4-difluorobenzyl) -N- (2,2-dimethoxyethyl) -4-toluenesulfonamide 129e (3500 mg,7.5 mmol) was dissolved in ultra dry dichloromethane (20 mL), and then the solution was slowly added to a mixture of aluminum trichloride (10026mg, 75.2mmol) in dichloromethane (20 mL) under nitrogen at 0 ℃. The reaction solution was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was gradually added to ice water to quench, followed by extraction with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (dichloromethane/petroleum ether = 80%) to give 8-bromo-6,7-difluoroisoquinoline 129f (1062 mg, orange solid) in yield: and 52 percent.
MS m/z(ESI)：244.0[M+1] + 。
The sixth step
Preparation of 6,7-difluoro-isoquinoline-8-carboxylic acid methyl ester
8-bromo-6,7-difluoroisoquinoline 129f (1062mg, 4.4 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (750mg, 1.10mmol) and triethylamine (896mg, 8.9mmol) were dissolved in methanol (20 mL), and then the reaction solution was stirred at 100 ℃ for 16 hours under an atmosphere of carbon monoxide (1.5 MPa). After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (ethyl acetate/petroleum ether = 20%) to give 6,7-difluoro isoquinoline-8-carboxylic acid methyl ester 129g (600 mg, grey solid), yield: 61 percent.
MS m/z(ESI)：224.1[M+1] + 。
Seventh step
Preparation of (6,7-Difluoroisoquinolin-8-yl) methanol
129g (600mg, 2.7 mmol) of methyl 6,7-difluoroisoquinoline-8-carboxylate was dissolved in anhydrous tetrahydrofuran (10 mL) and then a solution of lithium aluminum hydride (1.0M in tetrahydrofuran) (3 mL) was added dropwise to the solution at 0 ℃. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give (6,7-difluoroisoquinolin-8-yl) methanol 129h (300 mg, grey solid) yield: 56 percent.
MS m/z(ESI)：196.1[M+1] + 。
The eighth step
Preparation of 8- (bromomethyl) -6,7-difluoroisoquinoline
(6,7-Difluoroisoquinolin-8-yl) methanol 129h (300mg, 1.53mmol) was dissolved in dichloromethane (5 mL) and phosphorus tribromide (0.5 mL) was added dropwise to the solution at 0 ℃. The reaction mixture was stirred at 25 ℃ for 2 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 8- (bromomethyl) -6,7-difluoroisoquinoline 129i (180 mg, yellow solid), yield: 46 percent.
MS m/z(ESI)：258.0[M+1] + 。
The ninth step
Preparation of 6,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) isoquinoline
8- (bromomethyl) -6,7-difluoroisoquinoline 129i (100mg, 0.39mmol), 4-fluoro-2-methoxy-5-nitroaniline (250mg, 1.40mmol), potassium carbonate (1011mg, 5.4 mmol) and potassium iodide (52mg, 0.21mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 50%) to give 6,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) isoquinoline 129j (55 mg, yellow solid), yield: 40 percent.
MS m/z(ESI)：365.1[M+1] + 。
The tenth step
Preparation of 5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline
6,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) isoquinoline 129j (55mg, 0.15mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (650mg, 9.4 mmol) and saturated ammonium chloride solution (5 mL) to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline 129k (50 mg, yellow solid) yield: 96 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：335.1[M+1] + 。
The eleventh step
Preparation of 4- (3- (5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- ((6,7-Difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline 129k (50mg, 0.13mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (686mg, 2.0 mmol) and triethylamine (162mg, 1.52mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 129l (65 mg, yellow solid) of 4- (3- (5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester, yield: 82 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：576.2[M+1] + 。
Twelfth step
Preparation of 3- (5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Is provided with
Dimethyl 4- (3- (5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 129l (65mg, 0.11mmol) was dissolved in tetrahydrofuran/water =1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative purification (FA) to give 3- (5- ((6,7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-057 (12 mg, yellow solid) in yield: 20 percent.
MS m/z(ESI)：530.0[M+1] + 。
HPLC:95.84％(214nm),91.20％(254nm).
1 H NMR(400MHz,DMSO-d6)δ14.52(s,1H),12.00(s,1H),9.58(s,1H),8.61(d,J＝5.6Hz,1H),8.16-8.12(m,1H),7.91(d,J＝5.6Hz,1H),7.42-7.30(m,2H),7.18(d,J＝11.6Hz,1H),5.59(s,2H),3.80(s,3H).
Example 58
Preparation of 3- { 2-fluoro-5- [ (7-fluoroquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-058)
Figure BDA0003608368340001191
First step of
Preparation of 7-fluoro-8-methylquinoline
3-fluoro-2-methylaniline 130a (2.50g, 20.00mmol) and sodium iodide (450mg, 3.00mmol) were dissolved in concentrated sulfuric acid (6.8 mL). After the reaction flask was stirred at 140 ℃ for 0.5 hour, glycerol (2.12g, 20.00mmol) was slowly added. The reaction mixture was reacted at 140 ℃ for 2 hours. After completion of the reaction, the reaction solution was returned to room temperature, poured slowly into ice water, adjusted to pH =8-1 with saturated sodium carbonate solution, and extracted with ethyl acetate (3 × 100 mL). Then, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 90/10) to obtain 7-fluoro-8-methylquinoline 130b (1.10 g, colorless oily liquid) in 31% yield.
MS m/z(ESI)：162.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.96(dd,J＝4.0,1.6Hz,1H),8.13(dd,J＝8.4,1.6Hz,1H),7.66(dd,J＝9.2,6.0Hz,1H),7.41-7.29(m,2H),2.72(d,J＝2.4Hz,3H).
Second step of
Preparation of 8- (bromomethyl) -7-fluoroquinoline
7-fluoro-8-methylquinoline 130b (500mg, 3.10 mmol) was dissolved in carbon tetrachloride (10 mL), and benzoyl peroxide (150mg, 3.10 mmol) and N-bromosuccinimide (662mg, 3.72mmol) were added at room temperature. The reaction mixture was reacted at 85 ℃ for 4 hours. After completion of the reaction, the reaction mixture was concentrated to remove carbon tetrachloride, diluted with dichloromethane (15 mL), washed with water (5 mL) and saturated brine (2X 5 mL) in this order, the organic phases were combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 50/50) to give 8- (bromomethyl) -7-fluoroquinoline 130c (670 mg, yellow solid) in a yield of 90%.
MS m/z(ESI)：240.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.04(d,J＝2.6Hz,1H),8.17(dd,J＝8.0,1.2Hz,1H),7.86-7.78(m,1H),7.49-7.41(m,1H),7.40-7.32(m,1H),5.25(d,J＝1.6Hz,2H).
The third step
Preparation of 7-fluoro-8- (5-fluoro-2-methoxy-4-nitrophenoxymethyl) quinoline
8- (bromomethyl) -7-fluoroquinoline 130c (100mg, 0.42mmol) was dissolved in acetonitrile (5 mL), 4-fluoro-2-methoxy-5-nitrophenol (94mg, 0.50mmol), potassium carbonate (174mg, 1.26mmol), and potassium iodide (28mg, 0.17mmol) were sequentially added, and the reaction solution was reacted at 65 ℃ for 1 hour. After completion of the reaction, water (2 mL) was added, and the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 7-fluoro-8- (5-fluoro-2-methoxy-4-nitrophenoxymethyl) quinoline 130d (85 mg, yellow solid) in 59% yield.
MS m/z(ESI)：347.1[M+1] + 。
The fourth step
Preparation of 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyaniline
7-fluoro-8- (5-fluoro-2-methoxy-4-nitrophenoxymethyl) quinoline 130d (85mg, 0.25mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (137mg, 2.46mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyaniline 130e (80 mg, brown solid) in 95% yield.
MS m/z(ESI)：317.1[M+1] + 。
The fifth step
Preparation of 4- [ ({ 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyaniline 130e (80mg, 0.25mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (158mg, 0.50mmol) were dissolved in tetrahydrofuran (5 mL). Then, triethylamine (77mg, 0.75mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction solution was not subjected to any post-treatment to give a stock solution of 4- [ ({ 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 130f.
MS m/z(ESI)：558.2[M+1] + 。
The sixth step
Preparation of 3- { 2-fluoro-5- [ (7-fluoroquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 130f (5 mL stock solution) was dissolved in methanol with lithium hydroxide (13mg, 0.53mmol): water =3:1 (4 mL). The reaction mixture was reacted at room temperature for 2 hours. After the reaction was finished, the reaction mixture was directly concentrated and purified by a reverse column (water: acetonitrile =20 = 80) to give 3- { 2-fluoro-5- [ (7-fluoroquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-058 (75.60 mg, yellow solid) in 97.00% yield.
MS m/z(ESI)：512.2[M+1] + 。
HPLC:98.27％(214nm),94.14％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.33(s,1H),9.01(dd,J＝4.0,1.6Hz,1H),8.49(dd,J＝8.4,1.6Hz,1H),8.20(dd,J＝9.2,6.4Hz,1H),7.67(t,J＝9.2Hz,1H),7.60(dd,J＝8.4,4.4Hz,1H),7.36(d,J＝6.4Hz,1H),7.06(d,J＝11.6Hz,1H),6.66-6.58(m,1H),5.60–5.50(m,2H),3.72(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-111.54,-129.40.
Example 59
Preparation of 3- {5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-059)
Figure BDA0003608368340001211
First step of
Preparation of 1,2-difluoro-3-methyl-4-nitrobenzene
Adding HNO 3 (65%, 6.80g, 0.07mol) in H 2 SO 4( 15 mL), the mixture was stirred at 0 ℃ for 10min, 1,2-difluoro-3-methylbenzene 131a (6.00g, 0.05mol) was then slowly added, and stirring was continued for 10min. After the reaction was completed, the reaction solution was quenched by pouring it into ice water, extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 4/1) to obtain 1,2-difluoro-3-methyl-4-nitrobenzene 131b (4.80 g, white oil), yield: and 47 percent.
1 H NMR(400MHz,d 6 -DMSO)δ7.98-7.94(m,1H),7.58(dd,J＝17.6,9.2Hz,1H),2.45(d,J＝2.4Hz,3H).
Second step of
Preparation of 3,4-difluoro-2-methylaniline
1,2-difluoro-3-methyl-4-nitrobenzene 131b (4.80g, 0.0277mol) was dissolved in NH 4 To a Cl/MeCN (20 mL) solution, iron powder (31.02g, 0.5539mol) was added and stirred at 25 ℃ for 1 hour. The reaction was filtered and concentrated, and the residue was isolated and purified on silica gel column (petroleum ether/ethyl acetate = 2/1) to give 3,4-difluoro-2-methylaniline 131c (3 g, brown oil), yield: 56 percent.
MS m/z(ESI)：144.1[M+1] + 。
The third step
Preparation of 6,7-difluoro-8-methylquinoline
3,4-difluoro-2-methylaniline 131c (1.40g, 0.0098mol) and NaI (0.73g, 0.0049mol) were dissolved in H 2 SO 4( 15 mL), after oil bath Wen was raised to 140 ℃, glycerol (5.42g, 0.0588mol) was slowly added and stirring was continued for 3 hours, after the reaction was finished, the reaction solution was poured into ice-methanol to quench, and was filtered again, the pH of the filtrate was adjusted to =9 after spin-drying, and the filtrate was extracted with ethyl acetate (3 × 50 mL), the organic phases were combined and dried with anhydrous sodium sulfate, and the residue obtained after concentration was separated and purified with a silica gel column (petroleum ether/ethyl acetate = 6/1) to obtain 6,7-difluoro-8-methylquinoline 131d (120 mg, pale yellow solid) in yield: 6 percent.
MS m/z(ESI)：180.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.93(d,J＝4.0Hz,1H),8.10(d,J＝8.0Hz,1H),7.44-7.37(m,2H),2.76(d,J＝2.8Hz,3H).
The fourth step
Preparation of 8- (bromomethyl) -6,7-difluoroquinoline
6,7-difluoro-8-methylquinoline 131d (110mg, 0.61mmol), BPO (74mg, 0.31mmol) and NBS (153mg, 0.86mmol) were dissolved in CCl 4( 6 mL) of the solution, and the mixture was stirred at 85 ℃ for 5 hours under a nitrogen atmosphere. After the reaction was completed, the reaction mixture was directly concentrated, and the residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 8- (bromomethyl) -6,7-difluoroquinoline 131e (106 mg, white solid), yield: 60 percent.
MS m/z(ESI)：257.9[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.01(s,1H),8.14(dd,J＝8.0,6.8Hz,2H),7.56-7.48(m,3H),5.24-5.23(m,2H).
The fifth step
Preparation of 6,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline
8- (bromomethyl) -6,7-difluoroquinoline 131e (86mg, 0.33mmol), 4-fluoro-2-methoxy-5-nitrophenol (75mg, 0.40mmol), potassium carbonate (92mg, 0.67mmol) and potassium iodide (83mg, 0.50mmol) were dissolved in acetonitrile (10 mL). The mixture was reacted at 80 ℃ for 2 hours. After the reaction was completed, extraction was performed with dichloromethane (3 × 50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and after concentration, the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 6,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 131f (100 mg, yellow solid) in yield: 65 percent.
MS m/z(ESI)：365.0[M+1] + 。
The sixth step
Preparation of 5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
6,7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 131f (80mg, 0.2196mmol) was dissolved in NH 4 To a Cl/MeCN (4 mL) solution, iron powder (246mg, 4.39mmol) was added and stirred at 25 ℃ for 1 hour. The reaction was filtered and concentrated to give 5- [ (6,7-difluoroquinolin-8-yl) methoxy]131g (80 mg, brown solid) of 2-fluoro-4-methoxyaniline, yield: 76%.
MS m/z(ESI)：335.1[M+1] + 。
Step seven
Preparation of dimethyl 4- [ ({ {5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
Prepare for
5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 131g (70mg, 0.21mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (351mg, 1.05mmol) were dissolved in tetrahydrofuran (10 mL), then triethylamine (117mg, 2.10mmol) was added, and the mixture was stirred at 25 ℃ for 16 hours.
MS m/z(ESI)：576.1[M+1] + 。
Eighth step
Preparation of 3- {5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ {5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 131h (10 mL stock solution) was dissolved in methanol with lithium hydroxide (50mg, 2.10mmol): water =3:1 (4 mL). The mixture was reacted at 25 ℃ for 1 hour. After the reaction was completed, the reaction mixture was directly concentrated and preliminarily purified by reverse column (water: acetonitrile = 3:2) to prepare 3- {5- [ (6,7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-059 (34.68 mg, white solid) with a yield of 30%.
MS m/z(ESI)：530.0[M+1] + 。
HPLC:97.11％(214nm),97.00％(254nm)。
1 H NMR(400MHz,d 6 -DMSO)δ12.03(s,1H),8.99(dd,J＝4.0,1.6Hz,1H),8.46(dd,J＝8.4,1.6Hz,1H),8.20(dd,J＝10.8,9.2Hz,1H),7.65(dd,J＝8.4,4.4Hz,1H),7.42-7.39(m,2H),7.13(d,J＝11.6Hz,1H),5.61(s,2H),3.77(s,3H).
Example 60
Preparation of 3- (5- ((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-060)
Figure BDA0003608368340001231
First step of
Preparation of 1,5-difluoro-2-methyl-3-nitrobenzene
2,4-difluorotoluene (1.00g, 7.80mmol) and nitric acid (740mg, 11.70mmol) were dissolved in sulfuric acid (4 mL), and the reaction was stirred at 0 ℃ for 1h. After completion of the reaction, the reaction mixture was quenched with sodium bicarbonate to pH >7 and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was separated and purified by silica gel column (petroleum ether/ethyl acetate = 9/1) to give 1,5-difluoro-2-methyl-3-nitrobenzene 132b (638 mg, yellow solid) in 46% yield.
1 H NMR(400MHz,CDCl 3 ):δ8.00(t,J＝7.8Hz,1H),6.98(dd,J＝10.4,9.0Hz,1H),2.33(s,3H)。
Second step of
Preparation of 3,5-difluoro-2-methylaniline
1,5-difluoro-2-methyl-3-nitrobenzene 132b (600mg, 3.42mmol), iron powder (580mg, 10.40mmol) and ammonium chloride (556mg, 10.40mmol) were dissolved in ethanol: water =1:1 (10 mL), and the reaction mixture was reacted at 80 ℃ for 3 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 9/1) to obtain 3,5-difluoro-2-methylaniline 132c (462 mg, gray solid) in 87% yield.
MS m/z(ESI)：144.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ6.72(dd,J＝10.8,9.4Hz,1H),6.58(dd,J＝9.6,8.0Hz,1H),3.34(s,2H),2.16(s,3H)。
The third step
Preparation of 5,7-difluoro-8-methylquinoline
3,5-difluoro-2-methylaniline 132c (400mg, 2.80mmol) and sodium iodide (47mg, 0.28mmol) were dissolved in sulfuric acid (8 mL), glycerol (326mg, 3.10mmol) was added at 140 ℃ and the reaction mixture was reacted at 140 ℃ for 6 hours. After completion of the reaction, methanol (20 mL) was added to the reaction mixture under ice-bath conditions, followed by filtration and concentration. Saturated sodium bicarbonate solution was then added to the reaction solution to pH >7 and extracted with ethyl acetate (3 × 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by reverse phase column (33% acetonitrile/water) to give 5,7-difluoro-8-methylquinoline 132d (112 mg, yellow solid) in 22% yield.
MS m/z(ESI)：180.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.97(d,J＝2.4Hz,1H),8.36(d,J＝8.6Hz,1H),7.56(dd,J＝8.6,4.0Hz,1H),7.28(t,J＝9.8Hz,1H),2.53(s,3H)。
The fourth step
Preparation of 8- (bromomethyl) -5,7-difluoroquinoline
5,7-difluoro-8-methylquinoline 132d (96mg, 0.53mmol), N-bromosuccinimide (114mg, 0.64mmol) and benzoyl peroxide (65mg, 0.27mmol) were dissolved in carbon tetrachloride (4 mL) and the reaction was reacted at 85 ℃ for 6h under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched by addition of water (10 mL) and extracted with ethyl acetate (3X 10 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by reverse phase column (48% acetonitrile/water) to give 8- (bromomethyl) -5,7-difluoroquinoline 132e (76 mg, yellow solid) in 54% yield.
MS m/z(ESI)：258.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.01(d,J＝4.0Hz,1H),8.73(dt,J＝8.8,1.6Hz,1H),7.83(tt,J＝6.4,4.2Hz,2H),5.22(dd,J＝36.2,1.8Hz,2H)。
The fifth step
Preparation of 5,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline
8- (bromomethyl) -5,7-difluoroquinoline 132e (66mg, 0.26mmol), 4-fluoro-2-methoxy-5-nitrophenol (57mg, 0.31mmol), and potassium carbonate (42mg, 0.31mmol) were dissolved in acetonitrile (10 mL), and the reaction solution was reacted at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was isolated and purified by silica gel column (petroleum ether/ethyl acetate = 3/2) to give 5,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline 132f (69 mg, white solid) in 73% yield.
MS m/z(ESI)：365.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.00(dd,J＝4.2,1.2Hz,1H),8.67(d,J＝8.6Hz,1H),7.99(d,J＝7.4Hz,1H),7.89-7.72(m,2H),7.28(d,J＝13.4Hz,1H),5.61(d,J＝1.6Hz,2H),3.83(s,3H)。
The sixth step
Preparation of 5- ((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline
5,7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline 132f (59mg, 0.12mmol), iron powder (45mg, 0.81mmol) and ammonium chloride (43mg, 0.81mmol) were dissolved in ethanol: water =1:1 (3 mL), and the reaction mixture was reacted at 60 ℃ for 3 hours. After completion of the reaction, the reaction mixture was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 132g of 5- ((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline (55 mg, grey solid) in 96% yield.
MS m/z(ESI)：335.1[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ8.99(d,J＝3.4Hz,1H),8.68(d,J＝8.6Hz,1H),7.88-7.69(m,2H),6.84(d,J＝12.6Hz,1H),6.63(d,J＝8.8Hz,1H),5.36(s,2H),3.61(s,3H)。
Seventh step
Preparation of dimethyl 4- (3- (5- (((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
5- ((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline 132g (55mg, 0.12mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (83mg, 0.25mmol) were dissolved in tetrahydrofuran (10 mL), triethylamine (33mg, 0.33mmol) was added to the reaction at room temperature, the reaction liquid was reacted at 25 ℃ for 16 hours, after the reaction was completed, water (20 mL) was added to the reaction liquid, and the reaction liquid was quenched, extracted with ethyl acetate (3X 20 mL), all organic phases were combined, anhydrous sodium sulfate was dried and concentrated, and the resultant residue was separated and purified with a silica gel column (petroleum ether/ethyl acetate = 3/2) to give 4- (3- (5- (((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl urea) thiophene-3425 zxh dimethyl ester (132 mg, 58 mg, 59% yield as a yellow solid.
MS m/z(ESI)：576.0[M+1] + 。
1 H NMR(400MHz,d 6 -DMSO)δ8.98(dd,J＝5.0,3.8Hz,2H),8.83(s,1H),8.69(d,J＝8.6Hz,1H),7.94(s,1H),7.86-7.72(m,3H),7.03(d,J＝12.6Hz,1H),5.42(d,J＝1.2Hz,2H),3.89(s,3H),3.83(s,3H),3.69(s,3H)。
Eighth step
Preparation of 3- (5- ((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- (((5,7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 132H (45mg, 0.08mmol) and lithium hydroxide (18mg, 0.78mmol) were dissolved in tetrahydrofuran: methanol water = 1.
MS m/z(ESI)：530.0[M+1] + 。
HPLC:99.61％(214nm),97.40％(254nm)。
1 H NMR(400MHz,d 6 -DMSO)δ11.99(s,1H),9.00-8.99(m,1H),8.67(d,J＝8.8Hz,1H),7.95-7.69(m,3H),7.35(d,J＝8.8Hz,2H),7.16(d,J＝11.6Hz,1H),5.40(s,2H),3.79(s,3H)。
Example 61
Preparation of 3- {5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-061)
Figure BDA0003608368340001251
First step of
1,5-difluoro-2-methyl-3-nitrobenzene 148a (5g, 39mmol) was dissolved in concentrated sulfuric acid (20 mL). Nitric acid (2.70g, 42.9 mmol) was then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then quenched with ice and adjusted to pH 7-8 with 1M sodium hydroxide solution. Extraction with dichloromethane and purification of the concentrated residue by column chromatography (petrol ether/ethyl acetate: 100/0) gave 1,5-difluoro-2-methyl-3-nitrobenzene 134b (3.8 g, yellow solid) in 51% yield.
1 H NMR(400MHz,CDCl 3 )δ8.00(t,J＝8.0Hz,1H),6.98(dd,J＝10.6,8.8Hz,1H),2.38-2.27(m,3H).
Second step of
Preparation of 3,5-difluoro-2-methylaniline
1,5-difluoro-2-methyl-3-nitrobenzene 134b (500mg, 2.88mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (241mg, 4.33mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 3,5-difluoro-2-methylaniline 134c (200 mg, yellow liquid) in 43% yield.
MS m/z(ESI)：144.0[M+1] + 。
The third step
N- (3,5-difluoro-2-methylphenyl) acetamide
3,5-difluoro-2-methylaniline 134c (4.5g, 31.4 mmol) was dissolved in dichloromethane (50 mL) and acetyl chloride (7.39g, 94.1mmol) and triethylamine (9.51g, 94.1mmol) were added dropwise at 25 ℃. The reaction mixture was reacted at 25 ℃ for 1 hour. The solvent was then spun dry, extracted with aqueous sodium bicarbonate solution and dichloromethane, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give N- (3,5-difluoro-2-methylphenyl) acetamide 134d (4.5 g, yellow solid) in 70% yield.
MS m/z(ESI)：186[M+1] + 。
The fourth step
N- (3,5-difluoro-2-methyl-6-nitrophenyl) acetamide
N- (3,5-difluoro-2-methylphenyl) acetamide 134d (5g, 27.0 mmol) was dissolved in concentrated sulfuric acid (10 mL). Nitric acid (1.87g, 29.7 mmol) was then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 1 hour. Then quenched with ice and adjusted to pH 7-8 with 1M sodium hydroxide solution. Extraction with dichloromethane and purification of the concentrated residue by column chromatography (petroleum ether/ethyl acetate: 100/0) gave N- (3,5-difluoro-2-methyl-6-nitrophenyl) acetamide 134e (4 g, yellow liquid) in 58% yield.
MS m/z(ESI)：231[M+1] + 。
The fifth step
Preparation of 3,5-difluoro-2-methyl-6-nitroaniline
N- (3,5-difluoro-2-methyl-6-nitrophenyl) acetamide 134e (2.5 g,10.9 mmol) was dissolved in ethanol (20 mL). Concentrated sulfuric acid (2 mL) was then added at 25 ℃. The reaction mixture was reacted at 90 ℃ for 4 hours. The solvent was then spun dry and quenched with ice and the pH adjusted to 7-8 with 1M sodium hydroxide solution. Extraction with dichloromethane and purification of the concentrated residue by column chromatography (petroleum ether/ethyl acetate: 60/40) gave 3,5-difluoro-2-methyl-6-nitroaniline 134f (2.1 g, yellow liquid) in 92% yield.
MS m/z(ESI)：189[M+1] + 。
The sixth step
Preparation of 4,6-difluoro-3-methylbenzene-1,2-diamine
3,5-difluoro-2-methyl-6-nitroaniline 134f (2.1g, 11.2mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (0.63g, 11.2mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 4,6-difluoro-3-methylbenzene-1,2-diamine 134g (1.5 g, red solid) in 76% yield.
MS m/z(ESI)：159.0[M+1] + 。
Seventh step
Preparation of 6,8-difluoro-5-methylquinoxaline
4,6-difluoro-3-methylbenzene-1,2-diamine 134g (1.5 g,9.5 mmol) was dissolved in tetrahydrofuran (9 mL). Then, hydrogen fluoride (4.5 mL) was added to the reaction solution, and the reaction mixture was reacted at 0 ℃ for 1 hour. The pH was then adjusted to 7-8 with saturated sodium bicarbonate solution and extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 6,8-difluoro-5-methylquinoxaline 134h (0.4 g, yellow solid) in 21% yield.
MS m/z(ESI)：191.0[M+1] + 。
Eighth step
Preparation of 5- (bromomethyl) -6,8-difluoroquinoxaline
6,8-difluoro-5-methylquinoxaline 134h (200mg, 1.11mmol) was dissolved in carbon tetrachloride (15 mL). Then, dibenzoyl peroxide (134mg, 0.55mmol) and N-bromosuccinimide (237mg, 1.33mmol) were added thereto at 25 ℃ and the reaction mixture was reacted at 85 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (bromomethyl) -6,8-difluoroquinoxaline 134i (250 mg, red solid) in 78% yield.
MS m/z(ESI)：259.0[M+1] + 。
The ninth step
Preparation of 6,8-difluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline
134i (300mg, 1.16mmol) and 4-fluoro-2-methoxy-5-nitrophenol (216mg, 1.116mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (320mg, 2.32mmol) and potassium iodide (288mg, 1.73mmol) were added, and the reaction mixture was reacted at 65 ℃ for 2 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 6,8-difluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 134j (330 mg, white solid) in 70% yield.
MS m/z(ESI)：366.0[M+1] + 。
The tenth step
5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyaniline
6,8-difluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 134j (150mg, 0.4 mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (22mg, 0.41mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyaniline 134k (70 mg, brown solid) in 46% yield.
MS m/z(ESI)：336.0[M+1] + 。
The eleventh step
Dimethyl 4- [ ({ {5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyaniline 134k (60mg, 0.18mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (60mg, 0.18mmol) were dissolved in tetrahydrofuran (10 mL), triethylamine (54mg, 0.53mmol) were then added, and the reaction liquid was reacted at 25 ℃ for 1 hour, the reaction liquid was not post-treated at all to give dimethyl 4- [ ({ {5- [ (3536 zft 3536-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-3926 zft dicarboxylate 148l 148.
MS m/z(ESI)：577.0[M+1] + 。
The twelfth step
Preparation of 3- {5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ {5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 148l (10 mL stock solution) was dissolved in methanol with lithium hydroxide (5mg, 0.13mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction is finished, the reaction product is directly concentrated, and after the initial purification by a reverse column (water: acetonitrile =5 = 95), the 3- {5- [ (6,8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-061 (2.06 mg, yellow solid) is obtained after preparation and purification, and the yield is 4%.
MS m/z(ESI)：531.0[M+1] + 。
HPLC:90.74％(214nm),91.01％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.53(s,1H),12.00(s,1H),9.08(dd,J＝26.4,1.6Hz,2H),8.01(s,1H),7.40-7.32(m,2H),7.12(d,J＝11.6Hz,1H),5.50(s,2H),3.76(s,3H).
Example 62
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-062)
Figure BDA0003608368340001281
First step of
Preparation of 4-methyl-3H-1,3-benzoxazole-2-thione
2-amino-3-methylphenol 137a (3.80g, 30.86mmol) was dissolved in anhydrous methanol (50 mL), and then potassium sulfamethyl acetoacetate (9.96g, 60.71mmol) was added. The reaction was stirred at 90 ℃ for 4 hours. After LCMS detection of reaction completion, the reaction was cooled to room temperature. The solvent was distilled off under reduced pressure and purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to give 4-methyl-3H-1,3-benzoxazole-2-thione 137b (5.0 g, white solid) as a product, yield: 88 percent.
MS m/z(ESI)：166.2[M+1] + 。
Second step of
Preparation of 2-chloro-4-methyl-1,3-benzoxazole
4-methyl-3H-1,3-benzoxazole-2-thione 137b (2.00g, 12.11mmol) was dissolved in dichloromethane (10 mL) and phosphorus oxychloride (10 mL) was added followed by phosphorus pentachloride (2.77g, 13.32mmol) next. The reaction was stirred at 100 ℃ for 16 hours. After LCMS detection of reaction completion, the reaction was cooled to room temperature. After the solvent was distilled off under reduced pressure, ice was added and the pH was adjusted to 8-9 by saturated sodium carbonate solution, followed by extraction with methylene chloride (3X 20 mL). After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure and then purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain a product 2-chloro-4-methyl-1,3-benzoxazole 137b (500 mg, light yellow solid), yield: 19 percent.
MS m/z(ESI)：168.2[M+1] + 。
The third step
Preparation of 2-methoxy-4-methyl-1,3-benzoxazole
4-methyl-3H-1,3-benzoxazole-2-thione 137c (220mg, 1.31mmol) was dissolved in methanol (10 mL) followed by the addition of sodium methoxide (142mg, 2.63mmol). The reaction was refluxed at 90 ℃ for 5 hours. After completion of the reaction monitored by LCMS, the reaction was allowed to cool to room temperature and the solvent was distilled off under reduced pressure. Purification on silica gel column (petroleum ether/ethyl acetate = 1/1) gave the product 2-methoxy-4-methyl-1,3-benzoxazole 137d (180 mg, white solid) in yield: and 76 percent.
MS m/z(ESI)：164.2[M+1] + 。
The fourth step
Preparation of 4- (bromomethyl) -2-methoxy-1,3-benzoxazole
Isoquinolin-8-ylcarbinol 137d (200mg, 1.23mmol) was dissolved in carbon tetrachloride (5 mL) followed by the addition of N-bromosuccinimide (305mg, 1.72mmol) and benzoyl peroxide (148mg, 0.61mmol). The reaction was stirred at 80 ℃ for 4h. After the reaction was completed, the solvent was distilled off under reduced pressure and purified by a silica gel column to obtain 4- (bromomethyl) -2-methoxy-1,3-benzoxazole 137e (130 mg, pale yellow solid) as a product, yield: 39 percent.
MS m/z(ESI)：242.0[M+1] + 。
The fifth step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1,3-benzoxazole
4- (bromomethyl) -2-methoxy-1,3-benzoxazole 137e (100mg, 0.41mmol) and 4-fluoro-2-methoxy-5-nitroaniline (77mg, 0.41mmol) were dissolved in acetonitrile (20 mL) followed by the addition of potassium carbonate (114mg, 0.83mmol). The reaction was stirred at 50 ℃ for 4h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), drying over anhydrous sodium sulfate, and the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 4/1) to give 137f (120 mg, yellow solid) of preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1,3-benzoxazole: 67%.
MS m/z(ESI)：349.2[M+1] + 。
The sixth step
Preparation of 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] aniline
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1,3-benzoxazole 137f (100mg, 0.29mmol) was dissolved in acetonitrile (10 mL) and saturated ammonium chloride (2 mL), followed by addition of iron powder (160mg, 2.87mmol). The reaction was stirred at 25 ℃ for 0.5h. After completion of the reaction iron powder was removed by filtration and then extracted with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 137g (80 mg, brown solid) of 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] aniline (yield: 79 percent.
MS m/z(ESI)：319.1[M+1] + 。
Step seven
Preparation of 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzooxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2,3-dicarboxylic acid dimethyl ester
137g (40mg, 0.13mmol) of 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] aniline was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (51mg, 0.15mmol) and triethylamine (38mg, 0.38mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. After completion of the reaction monitored by LCMS, the solvent was distilled off under reduced pressure and purified by a silica gel column to give preparation of compound 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2,3-dicarboxylic acid dimethyl ester 137h (60 mg, yellow solid), yield: 68 percent.
MS m/z(ESI)：539.1[M+1] + 。
Eighth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzoxazol-4-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Preparation of 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1,3-benzooxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2,3-dicarboxylic acid dimethyl ester 137h (60mg, 0.07mmol) was dissolved in tetrahydrofuran (5 mL) and water (1 mL) and then lithium hydroxide monohydrate (9mg, 0.21mmol) was added. The reaction was stirred at room temperature for 1h. After completion of the reaction the solvent was evaporated to give a crude product which was purified by preparative (0.1% formic acid/acetonitrile/water) to give 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-062 (10 mg, white solid) yield: 27 percent.
MS m/z(ESI)：532.1[M+1] + 。
HPLC:98.97％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.56(s,1H),11.91(s,1H),7.54(d,J＝8.2Hz,1H),7.43(d,J＝7.2Hz,1H),7.33(d,J＝7.2Hz,1H),7.27(dd,J＝10.4,5.2Hz,2H),7.13(d,J＝11.6Hz,1H),5.19(s,2H),4.14(s,3H),3.83(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-128.55.
Example 63
Preparation of 3- (5- ((5-chloro-7-fluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-063)
Figure BDA0003608368340001301
First step of
Preparation of 4-chloro-2-fluoro-6-nitrophenol
Fuming nitric acid (3.97g, 40.92mmol) was added dropwise to a solution of 4-chloro-2-fluorophenol 146a (5.00g, 34.10 mmol) in acetic acid (30 mL) at 0 ℃. After the completion of the dropwise addition, the reaction mixture was further stirred at 0 ℃ for 1 hour. The reaction mixture was slowly poured into water to precipitate a solid, filtered, washed with water and the filter cake was dried to give 4-chloro-2-fluoro-6-nitrophenol 146b (5.89 g, yellow solid) in yield: 90 percent.
1 H NMR(400MHz,CDCl 3 )δ10.37(s,1H),7.94(t,J＝2.3Hz,1H),7.45(dd,J＝9.5,2.5Hz,1H).
Second step of
Preparation of 4-chloro-2-fluoro-6-nitrophenyl triflate
4-chloro-2-fluoro-6-nitrophenol 146b (5.89g, 30.80mmol) was dissolved in dichloromethane (30 mL), triethylamine (4.67g, 46.20 mmol) was added, trifluoromethanesulfonic anhydride (8.69g, 30.80mmol) was added dropwise, and then stirring was carried out at 0 ℃ for 2 hours. After the reaction was completed, water quenching was added and extraction was performed with dichloromethane (3 × 100 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to obtain 4-chloro-2-fluoro-6-nitrophenyl trifluoromethanesulfonate 146c (9.54 g, white solid) in yield: 96 percent.
1 H NMR(400MHz,CDCl 3 )δ7.99(t,J＝2.2Hz,1H),7.63(dd,J＝8.8,2.5Hz,1H).
The third step
Preparation of 5-chloro-1-fluoro-2-methyl-3-nitrobenzene
To a solution of 4-chloro-2-fluoro-6-nitrophenyl triflate 146c (9.54g, 29.48mmol) in dioxane (60 mL) was added methylboronic acid (2.12g, 35.49mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (2.16g, 2.96mmol) and potassium carbonate (6.12g, 44.26mmol) and stirred at 90 ℃ for 8 hours. After the reaction was completed, water was added to quench and extraction was performed with ethyl acetate (3 × 100 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to obtain 5-chloro-1-fluoro-2-methyl-3-nitrobenzene 146d (1.81 g, white solid) in yield: 32 percent.
1 H NMR(400MHz,CDCl 3 )δ7.75(t,J＝1.7Hz,1H),7.33(dd,J＝8.7,2.1Hz,1H),2.44(d,J＝2.2Hz,3H).
The fourth step
Preparation of 5-chloro-3-fluoro-2-methylaniline
To a solution of 5-chloro-1-fluoro-2-methyl-3-nitrobenzene 146d (1.81g, 9.55mmol) in ethanol (10 mL) and water (10 mL) were added ammonium chloride (1.53g, 28.64mmol) and iron powder (1.60g, 28.64mmol), and the mixture was stirred at 80 ℃ for 1 hour. After the completion of the reaction, filtration was carried out while it was hot, the filtrate was extracted with ethyl acetate (3 × 50 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 5-chloro-3-fluoro-2-methylaniline 146e (1.00 g, colorless liquid), yield: 66 percent.
MS m/z(ESI)：160.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ6.48(dd,J＝9.2,1.6Hz,1H),6.45-6.43(m,1H),3.77(s,2H),2.01(d,J＝2.0Hz,3H).
The fifth step
Preparation of 5-chloro-7-fluoro-8-methylquinoline
To a solution of 5-chloro-3-fluoro-2-methylaniline 146e (1.00g, 6.30mmol) and sodium iodide (0.47g, 3.10 mmol) in concentrated sulfuric acid (20 mL) at 140 ℃ was slowly added glycerol (3.48g, 37.80mmol) and the reaction was stirred at 140 ℃ for 10 hours. After the reaction, the reaction solution was slowly added to methanol to quench, filtered, the filtrate was concentrated and then added with ammonia water to adjust the system to alkalinity, extracted with ethyl acetate (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain 5-chloro-7-fluoro-8-methylquinoline 146f (120 mg, yellow solid), yield: 10 percent.
MS m/z(ESI)：196.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.06(dd,J＝4.2,1.6Hz,1H),8.56(dd,J＝8.5,1.6Hz,1H),7.87(d,J＝9.5Hz,1H),7.70(dd,J＝8.5,4.2Hz,1H),2.60(d,J＝2.5Hz,3H).
The sixth step
Preparation of 8- (bromomethyl) -5-chloro-7-fluoroquinoline
To a solution of 5-chloro-7-fluoro-8-methylquinoline 146f (120mg, 0.61mmol) in carbon tetrachloride (10 mL) were added 2,2' -azobis (2-methylpropionitrile) (10mg, 0.06mmol) and N-bromosuccinimide (120mg, 0.67mmol), and the mixture was stirred at 80 ℃ for 3 hours. After the reaction was completed, water was added to quench, and the mixture was extracted with dichloromethane (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to give 146g of 8- (bromomethyl) -5-chloro-7-fluoroquinoline (104 mg, white solid) in yield: 62 percent.
MS m/z(ESI)：276.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.08(d,J＝4.2Hz,1H),8.57(dd,J＝8.5,1.6Hz,1H),7.55(dd,J＝8.5,4.2Hz,1H),7.50(d,J＝9.2Hz,1H),5.20(s,2H).
Seventh step
Preparation of 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline
To a solution of 4-fluoro-2-methoxy-5-nitrophenol (85mg, 2.47mmol) in acetonitrile (10 mL) were added potassium carbonate (78mg, 0.57mmol) and 146g of 8- (bromomethyl) -5-chloro-7-fluoroquinoline (104mg, 0.38mmol), and the reaction mixture was stirred at 70 ℃ for 2 hours. After the reaction was completed, water quenching was added, extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 4/1) to obtain 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 146h (135 mg, white solid) in yield: 93 percent.
MS m/z(ESI)：381.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.12(dd,J＝4.2,1.6Hz,1H),8.65(dd,J＝8.6,1.6Hz,1H),8.04(d,J＝8.9Hz,2H),7.77(dd,J＝8.6,4.2Hz,1H),7.25(d,J＝13.4Hz,1H),5.76(s,2H),3.83(s,3H).
Eighth step
Preparation of 5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
To a solution of 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 146h (135mg, 0.35mmol) in ethanol (5 mL) and water (5 mL) were added iron powder (59mg, 1.06mmol) and ammonium chloride (57mg, 1.06mmol) and the reaction mixture was stirred at 80 ℃ for 1 h. After the completion of the reaction, filtration was carried out while it was hot, the filtrate was extracted with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated and dried to give 5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 146i (110 mg, yellow solid), yield: 88 percent.
MS m/z(ESI)：351.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.02(dd,J＝4.2,1.6Hz,1H),8.55(dd,J＝8.6,1.6Hz,1H),7.52-7.48(m,2H),6.68(d,J＝8.8Hz,1H),6.62(d,J＝12.0Hz,1H),5.70(d,J＝1.6Hz,2H),3.71(s,3H).
The ninth step
Preparation of 4- [ ({ {5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
Triethylamine (39mg, 0.38mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (103mg, 0.31mmol) were added to a solution of 5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 146i (90mg, 0.26mmol) in tetrahydrofuran (5 mL), stirred at 25 ℃ for 4 hours, after the reaction was completed, water was added to quench, extracted with ethyl acetate (3X 10 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 4- [ ({ {5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 146j (126 mg, yellow solid) in 83% yield.
MS m/z(ESI)：592.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.10(dd,J＝4.2,1.6Hz,1H),8.97(s,1H),8.83(s,1H),8.65(
dd,J＝8.6,1.6Hz,1H),8.03(d,J＝9.4Hz,1H),7.94(s,1H),7.88(d,J＝8.0Hz,1H),7.75(dd,J＝8.6,4.2Hz,1H),6.99(d,J＝12.6Hz,1H),5.60(s,2H),3.90(s,3H),3.83(s,3H),3.66(s,3H).
The tenth step
Preparation of 3- {5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of dimethyl 4- [ ({ {5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 146j (106mg, 0.18mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (13mg, 0.54mmol), and the mixture was reacted at 25 ℃ for 2 hours. After the reaction was completed, 2N diluted hydrochloric acid was added dropwise to adjust pH =5-6, extracted with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- {5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-063 (35 mg, white solid), yield: 36 percent.
MS m/z(ESI)：545.9[M+1] + 。
HPLC:98.65％(214nm),98.73％(254nm).
1 H NMR(400MHz,DMSO-d6)δ9.10(dd,J＝4.2,1.6Hz,1H),8.65(dd,J＝8.6,1.6Hz,1H),8.04(d,J＝9.4Hz,1H),7.75(dd,J＝8.6,4.2Hz,1H),7.38(d,J＝7.4Hz,1H),7.18(s,1H),7.10(d,J＝11.6Hz,1H),5.55(s,2H),3.74(s,3H).
Example 64
Preparation of 3- { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-064)
Figure BDA0003608368340001321
First step of
Preparation of 5-fluoro-4-methyl-1,3-benzothiazol-2-amine
3-fluoro-2-methylaniline 147a (10 g, 0.08mol) and potassium thiocyanate (31.06g, 0.32mol) were dissolved in acetic acid (100 mL), stirred at room temperature for 30min, and then Br was added 2 (12.64g, 0.0791mol) was diluted with acetic acid (30 mL) and added dropwise to the above solution while cooling on ice. The reaction solution was reacted at room temperature for 5 hours. After the reaction was completed, the reaction was quenched with water, adjusted to pH =8-9 with sodium hydroxide solution, and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 99/1) to give 5-fluoro-4-methyl-1,3-benzothiazol-2-amine 147b (8 g, white solid) in 10% yield.
MS m/z(ESI)：183.1[M+1] + 。
Second step of
Preparation of 2-bromo-5-fluoro-4-methyl-1,3-benzothiazole
5-fluoro-4-methyl-1,3-benzothiazol-2-amine 147b (8g, 0.04mol) was dissolved in acetonitrile (100 mL). Cuprous bromide (2.5 g, 0.04mol) and tert-butyl nitrite (13.58g, 0.1317 mol) were added in this order. The reaction mixture was reacted at 50 ℃ for 3 hours. After the reaction was completed, the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 99/1) to give 2-bromo-5-fluoro-4-methyl-1,3-benzothiazole 147c (9 g, yellow solid) with a yield of 8%.
1 H NMR(400MHz,CDCl 3 )δ7.56(dd,J＝8.4,4.4Hz,1H),7.16(t,J＝9.2Hz,1H),2.62(d,J＝2.0Hz,3H).
The third step
Preparation of 2-bromo-4- (bromomethyl) -5-fluoro-1,3-benzothiazole
2-bromo-5-fluoro-4-methyl-1,3-benzothiazole 147c (300mg, 1.22mmol) was dissolved in carbon tetrachloride (10 mL) and benzoyl peroxide (148mg, 0.61mmol) and N-bromosuccinimide (304mg, 1.71mmol) were added at room temperature. The reaction solution was reacted at 85 ℃ for 3 hours. After the reaction, the reaction mixture was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 99/1) to give 2-bromo-4- (bromomethyl) -5-fluoro-1,3-benzothiazole 147d (100 mg, white solid) with a yield of 17.67%.
MS m/z(ESI)：323.9[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.72(dd,J＝8.8,4.8Hz,1H),7.22(t,J＝9.2Hz,1H),4.98(d,J＝0.8Hz,2H).
The fourth step
Preparation of 2-bromo-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
2-bromo-4- (bromomethyl) -5-fluoro-1,3-benzothiazole 147d (600mg, 1.84mmol) was dissolved in acetonitrile (30 mL), 4-fluoro-2-methoxy-5-nitrophenol (345mg, 1.84mmol), potassium carbonate (510mg, 3.69mmol) and potassium iodide (459mg, 2.77mmol) were sequentially added, and the reaction solution was reacted at 65 ℃ for 1 hour. After completion of the reaction, the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 2-bromo-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 147e (400 mg, yellow solid) in 20% yield.
MS m/z(ESI)：452.9(M+23)。
The fifth step
Preparation of 5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
2-bromo-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 147e (110mg, 0.25mmol) and methylbenzeneboronic acid (7mg, 0.12mmol) were dissolved in toluene/ethanol/water (2ml, 7. Then, potassium carbonate (105mg, 0.76mmol) and tetrakis (triphenylphosphine) palladium (88mg, 0.07mmol) were added thereto, and the reaction solution was subjected to microwave reaction at 80 ℃ for 1 hour under nitrogen protection. After completion of the reaction, the solvent was removed under reduced pressure, water (2 mL) was added, extraction was performed with ethyl acetate (3X 10 mL), and the mixture was washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 80/20) to give 5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 147f (80 mg, yellow solid) in 71% yield.
MS m/z(ESI)：353.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.14(s,1H),8.01(d,J＝7.2Hz,1H),7.95(dd,J＝8.8,4.8Hz,1H),7.32-7.27(m,1H),6.69(d,J＝12.4Hz,1H),5.75(d,J＝0.8Hz,2H),3.90(s,3H).
The sixth step
Preparation of 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline
5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 147f (160mg, 0.45mmol) was dissolved in acetonitrile (2 mL). Then, a saturated ammonium chloride solution (2 mL) and iron powder (508mg, 9.08mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 147g (135 mg, brown solid) of 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline in 64% yield.
MS m/z(ESI)：323.1[M+1] + 。
Seventh step
Preparation of dimethyl 4- [ ({ { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
147g (135mg, 0.42mmol) of 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (280mg, 0.83mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (423.78mg, 4.188mmol) was then added to the reaction solution, and the reaction mixture was reacted at 25 ℃ for 16 hours. The reaction solution was not subjected to any post-treatment to give a stock solution of 4- [ ({ { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 147h.
MS m/z(ESI)：564.0[M+1] + 。
Eighth step
Preparation of 3- { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 147h (5 mL stock solution) was dissolved in methanol with lithium hydroxide (50mg, 2.12mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and subjected to preliminary purification by reverse column (water: acetonitrile = 20) to prepare and purify 3- { 2-fluoro-5- [ (5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-064 (62.03 mg, light yellow solid) in 54% yield.
MS m/z(ESI)：518.1[M+1] + 。
HPLC:100％(214nm),97.01％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.55(s,1H),12.02(s,1H),9.50(s,1H),8.30(dd,J＝8.8,5.2Hz,1H),7.50(t,J＝9.2Hz,1H),7.40(t,J＝3.6Hz,2H),7.13(d,J＝11.6Hz,1H),5.45(s,2H),3.78(s,3H).
Example 65
Preparation of 3- {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-065)
Figure BDA0003608368340001341
First step of
Preparation of 1-benzoyl-3- (3-chloro-2-methylphenyl) thiourea
3-chloro-2-methylaniline 148a (10g, 70.6mmol) was dissolved in tetrahydrofuran (90 mL). Benzoyl isothiocyanate (23.04g, 141.2mmol) was then added at 25 ℃. The reaction mixture was reacted at 25 ℃ for 16 hours. Then spin-drying the solvent and then mixing with ethanol: water =4:1 (50 mL) was filtered and the residue was taken to give 1-benzoyl-3- (3-chloro-2-methylphenyl) thiourea 148b (20.2 mg, white solid) in 84% yield.
MS m/z(ESI)：305[M+1] + 。
Second step of
Preparation of (3-chloro-2-methylphenyl) thiourea
1-benzoyl-3- (3-chloro-2-methylphenyl) thiourea 148b (20g, 65.6 mmol) was dissolved in methanol: water =3:1 (60 mL). Potassium carbonate (27.20g, 196.8mmol) was then added at 25 ℃. The reaction mixture was reacted at 70 ℃ for 2 hours. The solvent was then dried by spinning, water was added and the residue was filtered to give (3-chloro-2-methylphenyl) thiourea 148c (11 g, white solid) in 75% yield.
MS m/z(ESI)：201[M+1] + 。
The third step
Preparation of 5-chloro-4-methyl-1,3-benzothiazol-2-amine
(3-chloro-2-methylphenyl) thiourea 148c (12g, 59.8mmol) was dissolved in chloroform (50 mL), and then liquid bromine (11.47g, 71.7mmol) was added dropwise at 25 ℃. The reaction mixture was reacted at 60 ℃ for 2 hours. The solvent was then spun dry, water was added and the residue filtered to give 5-chloro-4-methyl-1,3-benzothiazol-2-amine 148d (11 g, white solid) in 83% yield.
MS m/z(ESI)：199[M+1] + 。
The fourth step
Preparation of 5-chloro-4-methyl-1,3-benzothiazole
5-chloro-4-methyl-1,3-benzothiazol-2-amine 148d (1.3g, 0.006mol) was dissolved in 1,4-dioxane (20 mL). Thereafter, tert-butyl nitrite (2.01g, 0.011mol) was added thereto, and the reaction mixture was reacted at 90 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 60/40) to give 5-chloro-4-methyl-1,3-benzothiazole 148e (1.1 g, brown solid) in 83% yield.
MS m/z(ESI)：184[M+1] + 。
The fifth step
Preparation of 4- (bromomethyl) -5-chloro-1,3-benzothiazole
5-chloro-4-methyl-1,3-benzothiazole 148e (1.2g, 1.80mmol) was dissolved in carbon tetrachloride (15 mL). Then, dibenzoyl peroxide (0.79g, 3.2mmol) and N-bromosuccinimide [1.39g,7.8mmol ] were added thereto at 25 ℃ and the reaction mixture was reacted at 85 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 4- (bromomethyl) -5-chloro-1,3-benzothiazole 148e (0.9 g, red solid) in 48% yield.
MS m/z(ESI)：263.0[M+1] + 。
The sixth step
Preparation of 5-chloro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
4- (bromomethyl) -5-chloro-1,3-benzothiazole 148e (900mg, 3.43mmol) and 4-fluoro-2-methoxy-5-nitrophenol (769mg, 4.11mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (0.947g, 6.85mmol) and potassium iodide (0.853g, 5.14mmol) were added, and the reaction mixture was reacted at 65 ℃ for 2 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5-chloro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 148f (700 mg, yellow solid) in 50% yield.
MS m/z(ESI)：369.0[M+1] + 。
Seventh step
Preparation of 5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline
5-chloro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 148f (900mg, 2.44mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (408mg, 7.32mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 148g (700 mg, brown solid) of 5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline in 76% yield.
MS m/z(ESI)：339.0[M+1] + 。
Eighth step
Preparation of 4- [ ({ {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline 148g (200mg, 0.59mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (296mg, 0.88mmol) were dissolved in tetrahydrofuran (10 mL), then triethylamine (178mg, 1.77mmol) was added, and the reaction solution was reacted at 25 ℃ for 1 hour to obtain a raw solution of dimethyl 4- [ ({ {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 148h without any post-treatment.
MS m/z(ESI)：580.0[M+1] + 。
The ninth step
Preparation of 3- {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 148h (10 mL stock solution) was dissolved in methanol with lithium hydroxide (21mg, 0.078mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and subjected to preliminary purification by reverse column (water: acetonitrile = 5) to prepare and purify 3- {5- [ (5-chloro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-065 (96.43 mg, brown solid) in 47% yield.
MS m/z(ESI)：534.0[M+1] + 。
HPLC:96.25％(214nm),97.75％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.56(s,1H),12.01(s,1H),9.49(s,1H),8.30(d,J＝8.4Hz,1H),7.67(d,J＝8.8Hz,1H),7.44-7.37(m,2H),7.13(d,J＝11.6Hz,1H),5.52(s,2H),3.77(s,3H).
Example 66
Preparation of 3- (5- ((5-chloro-7-fluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-066)
Figure BDA0003608368340001371
First step of
Preparation of 2-amino-5-chloro-3-fluorobenzoic acid
To a solution of 149a (10.00g, 64.50mmol) of 2-amino-3-fluorobenzoic acid in N, N-dimethylformamide (60 mL) was added N-chlorobutyldiimide (10.34g, 77.40mmol), and the reaction was stirred at 25 ℃ for 16 hours. After completion of the reaction, water was added to quench and filtered. The filter cake was dried to give 149b 2-amino-5-chloro-3-fluorobenzoic acid (11.0 g, yellow solid) in: and 90 percent.
MS m/z(ESI)：190.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ7.53-7.51(m,1H),7.44(dd,J＝11.2,2.5Hz,1H).
Second step of
Preparation of methyl 2-amino-5-chloro-3-fluorobenzoate
To a solution of 149b (11.00g, 58.00mmol) of 2-amino-5-chloro-3-fluorobenzoic acid in N, N-dimethylformamide (100 mL) were added iodomethane (8.83g, 69.60mmol) and potassium carbonate (12.02g, 87.00mmol), and the reaction was stirred at 25 ℃ for 16 hours. After completion of the reaction, water was added to quench and filtered. The filter cake was dried to give methyl 2-amino-5-chloro-3-fluorobenzoate 149c (10.00 g, as a pink solid) in yield: 85 percent.
MS m/z(ESI)：204.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ7.53(s,1H),7.49(dd,J＝11.2,2.4Hz,1H),6.70(s,2H),3.83(s,3H).
The third step
Preparation of methyl 2-bromo-5-chloro-3-fluorobenzoate
To a solution of methyl 2-amino-5-chloro-3-fluorobenzoate 149c (10.00g, 49.10 mmol) in acetonitrile (60 mL) was added dropwise a 40% aqueous solution of hydrogen bromide (60 mL) and an aqueous solution of sodium nitrite (4.07g, 58.90mmol) under nitrogen protection at 0 ℃ to react for 10 minutes, and cuprous bromide (8.45g, 58.90mmol) was added in portions. After the addition was complete, the reaction was continued at 0 ℃ for 1 hour, after which time it was diluted with water and extracted with ethyl acetate (3X 100 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to give methyl 2-bromo-5-chloro-3-fluorobenzoate 149d (9.60 g, white solid), yield: 73 percent.
1 H NMR(400MHz,CDCl 3 )δ760-7.59(m,1H),7.29–7.27(m,1H),3.96-3.95(m,3H).
The fourth step
Preparation of (2-bromo-5-chloro-3-fluorophenyl) methanol
To a solution of methyl 2-bromo-5-chloro-3-fluorobenzoate 149d (9.76g, 36.50mmol) in tetrahydrofuran (60 mL) at 0 deg.C under nitrogen was added dropwise a solution of 1N diisobutylaluminum hydride in hexanes (40 mL) and the reaction was stirred at 0 deg.C for 2 hours. After completion of the reaction, water and saturated sodium bicarbonate solution were added to quench and filtered, and the filtrate was extracted with ethyl acetate (3 × 50 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give (2-bromo-5-chloro-3-fluorophenyl) methanol 149e (7.60 g, white solid) in yield: 87 percent.
1 H NMR(400MHz,CDCl 3 )δ7.34(s,1H),7.09(dd,J＝8.0,2.4Hz,1H),4.74(s,2H),2.11(s,1H).
The fifth step
Preparation of 2-bromo-5-chloro-3-fluorobenzaldehyde
To a solution of (2-bromo-5-chloro-3-fluorophenyl) methanol 149e (7.60g, 31.70mmol) in methylene chloride (60 mL) was added Dess-Martin reagent (17.48g, 41.20mmol), and the reaction was stirred at 25 ℃ for 0.5 hour. After completion of the reaction, it was quenched by addition of saturated aqueous sodium bisulfite and saturated sodium bicarbonate solution, and extracted with the filtrate with dichloromethane (3X 100 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to give 2-bromo-5-chloro-3-fluorobenzaldehyde 149f (6.50 g, white solid) in yield: 86 percent.
1 H NMR(400MHz,CDCl 3 )δ10.24-10.23(m,1H),7.66-7.62(m,1H),7.32(dt,J＝7.7,2.4Hz,1H).
The sixth step
Preparation of [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine
A solution of 2-bromo-5-chloro-3-fluorobenzaldehyde 149f (6.50g, 27.40mmol) and 2,2-dimethoxyethylamine (3.46g, 32.80mmol) in ethanol (60 mL) was stirred at 80 ℃ for 8 hours, followed by addition of sodium cyanoborohydride (2.07g, 32.80mmol) and acetic acid (2 mL) and reaction at 25 ℃ for 1 hour. After completion of the reaction, saturated aqueous ammonium chloride solution was added to quench, extraction was performed with ethyl acetate (3 × 100 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 149g (8.00 g, yellow oily substance) of [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine, yield: 89 percent.
MS m/z(ESI)：328.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.27-7.26(m,1H),7.07(dd,J＝8.0,2.4Hz,1H),4.50(t,J＝5.4Hz,1H),3.89(s,2H),3.39(s,6H),3.22(s,1H),2.76(d,J＝5.4Hz,2H).
Seventh step
Preparation of N- [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine
To a solution of 149g (8.60g, 26.30mmol) of [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] (2,2-dimethoxyethyl) amine and 149g (8.60g, 39.30mmol) of triethylamine (3.99g, 39.40mmol) in dichloromethane (80 mL) was added dropwise p-toluenesulfonyl chloride (5.52g, 28.90mmol) at 0 ℃ and reacted at 0 ℃ for 3 hours. After completion of the reaction, it was quenched by addition of water (100 mL), extracted with dichloromethane (3X 100 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 20/1) to give N- [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine 149h (11.00 g, white solid), yield: 87 percent.
MS m/z(ESI)：504.0(M+23)。
1 H NMR(400MHz,CDCl 3 )δ7.72(d,J＝8.4Hz,2H),7.34(d,J＝8.0Hz,2H),7.17(s,1H),7.04(dd,J＝8.0,2.4Hz,1H),4.51(s,2H),4.41(t,J＝5.4Hz,1H),3.29(d,J＝5.4Hz,2H),3.27(s,6H),2.45(s,3H).
Eighth step
Preparation of 8-bromo-5-chloro-7-fluoroisoquinoline
To a solution of aluminum trichloride (15.20g, 114.00mmol) in methylene chloride (30 mL) was added dropwise a solution of N- [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] -N- (2,2-dimethoxyethyl) -4-methyl-1-sulfonylamine (149 h) (11.00g, 22.80mmol) in methylene chloride (60 mL) at 0 ℃ and, after completion of the dropwise addition, the reaction was carried out at 0 ℃ for 8 hours. After completion of the reaction, the reaction solution was quenched by slowly dropping into ice water and extracted with dichloromethane (3X 100 mL). The organic phase was washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to give 8-bromo-5-chloro-7-fluoroisoquinoline 149i (2.85 g, white solid) with yield: and 48 percent.
MS m/z(ESI)：262.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.56-9.54(m,1H),8.80-8.78(m,1H),8.29-8.25(m,1H),8.04(d,J＝6.0Hz,1H).
The ninth step
Preparation of 5-chloro-7-fluoroisoquinoline-8-carboxylic acid methyl ester
To a solution of 8-bromo-5-chloro-7-fluoroisoquinoline 149i (1.00g, 3.80mmol) and triethylamine (580mg, 5.70mmol) in methanol (20 mL) was added [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) (280mg, 0.30mmol), and the mixture was reacted at 80 ℃ for 16 hours under 1atm of carbon monoxide. After completion of the reaction, it was quenched by addition of water (20 mL) and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give 5-chloro-7-fluoroisoquinoline-8-carboxylic acid methyl ester 149j (653 mg, white solid) in yield: 71 percent.
MS m/z(ESI)：240.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.53(s,1H),8.76(d,J＝5.8Hz,1H),8.26(d,J＝10.0Hz,1H),8.10(d,J＝5.8Hz,1H),4.05(s,3H).
The tenth step
Preparation of (5-chloro-7-fluoroisoquinolin-8-yl) methanol
To a solution of methyl 5-chloro-7-fluoroisoquinoline-8-carboxylate 149j (353mg, 1.47mmol) in tetrahydrofuran (10 mL) was added dropwise a 1N solution of lithium aluminum hydride in tetrahydrofuran (2.2 mL) at 0 ℃ under a nitrogen blanket, and the mixture was reacted at 0 ℃ for 2 hours. After completion of the reaction, water was added to the reaction solution to quench (5 mL), and the mixture was extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give (5-chloro-7-fluoroisoquinolin-8-yl) methanol 149k (300 mg, yellow solid) in yield: 96 percent.
MS m/z(ESI)：212.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.70(s,1H),8.69(d,J＝5.8Hz,1H),8.08(d,J＝9.6Hz,1H),8.02(dd,J＝5.8,0.8Hz,1H),5.57(t,J＝5.6Hz,1H),5.01(dd,J＝5.2,2.0Hz,2H).
The eleventh step
Preparation of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline
After adding phosphine tribromide (687 mg, 2.54mmol) dropwise to a solution of (5-chloro-7-fluoroisoquinolin-8-yl) methanol 149k (358mg, 1.69mmol) in dichloromethane (10 mL) at 0 ℃, the reaction was carried out at 0 ℃ for 2 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added to adjust pH =6-7, and extraction was performed with ethyl acetate (3 × 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to give 149l of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline (380 mg, yellow oil) in yield: the content of the organic solvent is 82%,
MS m/z(ESI)：276.0[M+1] + 。
twelfth step
Preparation of 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
To a solution of 4-fluoro-2-methoxy-5-nitrophenol (311mg, 1.66mmol) and potassium carbonate (287mg, 2.08mmol) in acetonitrile (10 mL) was added 149l (380mg, 1.38mmol) of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline, and the mixture was reacted at 70 ℃ for 3 hours. After completion of the reaction, it was quenched by addition of water (10 mL) and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 149m (187 mg, white solid) in yield: 35 percent.
MS m/z(ESI)：381.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.66(s,1H),8.74(d,J＝5.8Hz,1H),8.19(d,J＝9.6Hz,1H),8.04(dd,J＝19.8,6.6Hz,2H),7.29(d,J＝13.4Hz,1H),5.75(s,2H),3.83(s,3H).
Thirteenth step
Preparation of 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline to a solution of 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline 149m (157mg, 0.41mmol) in ethanol (4 mL) and water (4 mL) were added iron powder (69mg, 1.24mmol) and ammonium chloride (66mg, 1.24mmol) and reacted at 80 ℃ for 1 hour. After completion of the reaction, extraction with ethyl acetate (3X 20 mL), drying of the organic phase over anhydrous sodium sulfate and filtration, and concentration and drying of the filtrate gave 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 149n (104 mg, yellow solid) in yield: 72 percent.
MS m/z(ESI)：351.1[M+1] + 。
Fourteenth step
Preparation of 4- [ ({ {5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
To a solution of 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 149n (100mg, 0.29mmol) and triethylamine (43mg, 0.43mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (115mg, 0.34mmol), reacted at 25 ℃ for 5 hours, quenched after completion of the reaction by addition of water (5 mL), extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give 4- [ ({ 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester o (110 mg, 149% yield: 65%.
MS m/z(ESI)：592.1[M+1] + 。
The fifteenth step
Preparation of 3- {5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid to a solution of 4- [ ({ {5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 149o (100mg, 0.17mmol) in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL), lithium hydroxide (12mg, 0.51mmol) was added and the reaction was carried out at 25 ℃ for 3 hours. After completion of the reaction, 1M diluted hydrochloric acid was added dropwise to adjust pH =4-5, extracted with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- {5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-066 (44 mg, white solid), yield: 48 percent.
MS m/z(ESI)：546.0[M+1] + 。
HPLC:98.48％(214nm),96.00％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ9.67(s,1H),8.74(d,J＝6.0Hz,1H),8.16(d,J＝9.6Hz,1H),8.08(dd,J＝6.0,1.0Hz,1H),7.36(d,J＝7.4Hz,1H),7.27(s,1H),7.16(d,J＝11.6Hz,1H),5.55(s,2H),3.79(s,3H).
Example 67
Preparation of 3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-067)
Figure BDA0003608368340001401
First step of
Preparation of N- ((5-chloro-2-methylphenyl) aminomethylsulfonyl) benzamide
5-chloro-2-methylaniline 150a (2.0 g, 14.1mmol) was dissolved in tetrahydrofuran (100 mL), followed by addition of benzoyl isothiocyanate (3.45g, 21.1mmol). The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, concentration was carried out to obtain the compound N- ((5-chloro-2-methylphenyl) aminomethylmethanesulfonyl) benzamide 150b (4 g, yellow solid) in a yield of 89%.
MS m/z(ESI)：305.0[M+1] + 。
Second step of
Preparation of 1- (5-chloro-2-methylphenyl) thiourea
N- ((5-chloro-2-methylphenyl) aminomethylcarbamoyl) benzamide 150b (4 g, 13.1mmol) was dissolved in methanol (60 mL), an aqueous solution (20 mL) of sodium carbonate (2.78g, 26.2mmol) was added, and the reaction solution was stirred at 70 ℃ for 2 hours. After completion of the reaction, concentration was carried out to obtain a crude product, which was then extracted with dichloromethane (3X 30 mL), and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and distilled under reduced pressure to remove the solvent, thereby obtaining compound 1- (5-chloro-2-methylphenyl) thiourea 150c (2.57 g, yellow oil) in 98% yield.
MS m/z(ESI)：201.1[M+1] + 。
The third step
Preparation of 7-chloro-4-methylbenzo [ d ] thiazol-2-amine
1- (5-chloro-2-methylphenyl) thiourea 150c (2.57g, 12.8 mmol) was dissolved in chloroform (100 mL), liquid bromine (2.04g, 12.8 mmol) was added, and the reaction solution was stirred at 70 ℃ for 2 hours. After the reaction was completed, an aqueous sodium thiosulfate solution (50 mL) was added, the organic phase was separated after washing with water, the aqueous phase was extracted with dichloromethane (3X 30 mL), the organic phase was separated, the combined organic phases were dried over anhydrous sodium sulfate, and concentrated to give compound 7-chloro-4-methylbenzo [ d ] thiazol-2-amine 150d (1.8 g, white solid) in 71% yield.
MS m/z(ESI)：199.0[M+1] + 。
The fourth step
Preparation of 7-chloro-4-methylbenzo [ d ] thiazole
7-Chlorobenzene-4-methyl [ d ] thiazol-2-amine 150d (1.8g, 9.06mmol) was dissolved in tetrahydrofuran (100 mL), tert-butyl nitrite (2.82g, 27.3mol) was added, and the reaction solution was stirred at 70 ℃ for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column (petroleum ether/ethyl acetate = 20/1) to obtain 7-chloro-4-methylbenzo [ d ] thiazole 150e (1.3 g, yellow solid) in 74% yield.
MS m/z(ESI)：184.0[M+1] + 。
The fifth step
Preparation of 4- (bromomethyl) -7-chlorobenzo [ d ] thiazole
7-chloro-4-methylbenzo [ d ] thiazole 150e (400mg, 2.18mmol) was dissolved in carbon tetrachloride (50 mL), and then N-bromosuccinimide (426.4mg, 2.39mmol) and azobisisobutyronitrile (35.8mg, 0.218mmol) were added, respectively, and the reaction solution was stirred at 80 ℃ for 3 hours. After the completion of the reaction, the reaction solution was concentrated, and the crude product was purified by silica gel column (petroleum ether/ethyl acetate = 20/1) to obtain 4- (bromomethyl) -7-chlorobenzo [ d ] thiazole 150f (430 mg, white solid) in 71% yield.
MS m/z(ESI)：263.9[M+1] + 。
The sixth step
Preparation of 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole
4- (bromomethyl) -7-chlorobenzo [ d ] thiazole 150f (200mg, 0.76mmol), 4-fluoro-2-methoxy-5-nitroaniline (171mg, 0.91mmol), potassium carbonate (211mg, 1.52mmol) and potassium iodide (63mg, 0.38mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 23%) to give 150g of 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole (210 mg, white solid), yield: 71 percent.
MS m/z(ESI)：369.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.57(s,1H),7.86(d,J＝7.4Hz,1H),7.70(t,J＝5.6Hz,2H),7.29(d,J＝13.4Hz,1H),5.66(s,2H),3.90(s,3H).
Seventh step
Preparation of 5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline
150g (85mg, 0.23mmol) of 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole, iron powder (38mg, 0.69mmol) and ammonium chloride (37mg, 0.69mmol) were dissolved in ethanol: water =1:1 (12 mL), and the reaction mixture was stirred at 60 ℃ for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was quenched with water (20 mL), dichloromethane (3 × 20 mL) was extracted, and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline after 150h (80 mg, brown solid), yield: 97 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：339.1[M+1] + 。
Eighth step
Preparation of 4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline 150h (80mg, 0.24mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (95mg, 0.28mmol) and triethylamine (47mg, 0.47mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was finished, rotary evaporation was performed to obtain a crude product of 4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 150i (70 mg, yellow solid), yield: 48 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：580.0[M+1] + 。
The ninth step
Preparation of 3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 150i (60mg, 0.10 mmol) was dissolved in tetrahydrofuran/water =1:1 (10 mL), followed by lithium hydroxide (8mg, 0.31mmol). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was adjusted to pH <7 with 1M HCl, washed with water (10 mL), extracted with ethyl acetate (3X 10 mL), the combined organic layers were washed with brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA) to give 3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-067 (43.03 mg, yellow solid), yield: and 76 percent.
MS m/z(ESI)：534.0[M+1] + 。
HPLC:100％(214nm),98.48％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.56(s,1H),11.99(s,1H),9.51(s,1H),7.71(d,J＝10.4Hz,2H),7.37(d,J＝8.4Hz,2H),7.16(d,J＝11.6Hz,1H),5.49(s,2H),3.84(s,3H).
Example 68
Preparation of 3- { 2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-068)
Figure BDA0003608368340001431
First step of
Preparation of 1-benzoyl-3- (5-fluoro-2-methylphenyl) thiourea
5-fluoro-2-methylaniline 151a (2g, 169mol) was dissolved in tetrahydrofuran (20 mL), and benzoyl isothiocyanate (5.22g, 32mmol) was added dropwise to the solution. The reaction solution was stirred at 25 ℃ for 16 hours under air protection. After the reaction was complete, the solvent was removed in vacuo and the solid was washed with a mixed solvent of ethanol: petroleum ether =4:1 (50 mL) and filtered. Filter cake was 1-benzoyl-3- (5-fluoro-2-methylphenyl) thiourea 151b (2.8 g, white solid), yield: 59 percent.
MS m/z(ESI)：289.0[M+1] + 。
Second step of
Preparation of (5-fluoro-2-methylphenyl) thiourea
1-benzoyl-3- (5-fluoro-2-methylphenyl) thiourea 151b (2.8g, 9.7mmol) and potassium carbonate (2.68g, 19.4mmol) were dissolved in methanol: water =4:1 (30 mL), and the reaction solution was stirred at 70 ℃ for 2 hours. After the reaction was complete, the solvent was removed in vacuo and then water (30 mL) was added. The solid was collected by filtration and further washed with water (30 mL). Filter cake was (5-fluoro-2-methylphenyl) thiourea 151c (1.8 g, white solid), yield: 96 percent.
MS m/z(ESI)：185.1[M+1] + 。
The third step
Preparation of 7-fluoro-4-methyl-1,3-benzothiazol-2-amine
(5-fluoro-2-methylphenyl) thiourea 151c (1.8g, 0.9mmol) was dissolved in chloroform (30 mL) and liquid bromine (2.35g, 14.7mmol) was added dropwise at room temperature. The reaction solution was stirred at 70 ℃ for 18 hours. After the reaction was completed, the solvent was removed in vacuo, and the solid was dissolved in a mixed solvent of dichloromethane: methanol = 1: 1 (30 mL), to which an aqueous solution of sodium thiosulfate (20 mL) was added. The solvent was then removed in vacuo and water (10 mL) was added. The solid was collected by filtration and purified by silica gel column (ethyl acetate/petroleum ether = 32%) to give 7-fluoro-4-methyl-1,3-benzothiazol-2-amine 151d (890 mg, yellow solid), yield: 49 percent.
MS m/z(ESI)：183.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.08-7.04(m,1H),6.84-6.74(m,1H),5.48(s,2H),2.50(s,3H).
The fourth step
Preparation of 7-fluoro-4-methyl-1,3-benzothiazole
7-fluoro-4-methyl-1,3-benzothiazol-2-amine 151d (890mg, 4.88mmol) was dissolved in 1,4-dioxane (30 mL), and tert-butyl nitrite (1000mg, 9.77mmol) was then added dropwise to the solution at room temperature. The mixture was stirred at 90 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (30 mL), extracted with ethyl acetate (3X 30 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 5%) to give 7-fluoro-4-methyl-1,3-benzothiazole 151e (440 mg, yellow liquid), yield: and 53 percent.
MS m/z(ESI)：168.2(M+H)。
1 H NMR(400MHz,CDCl 3 )δ9.01(d,J＝1.2Hz,1H),7.32-7.27(m,1H),7.10-7.05(m,1H),2.75(s,3H).
The fifth step
Preparation of 4- (bromomethyl) -7-fluoro-1,3-benzothiazole
7-fluoro-4-methyl-1,3-benzothiazole 151e (440mg, 2.63mmol), N-bromosuccinimide (562mg, 3.16mmol) and azobisisobutyronitrile (216mg, 2.16mmol) were dissolved in carbon tetrachloride (10 mL). The reaction solution was stirred at 85 ℃ for 16 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 5%) to give 4- (bromomethyl) -7-fluoro-1,3-benzothiazole 151f (270 mg, yellow solid) in yield: 41 percent.
MS m/z(ESI)：248.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.10(d,J＝1.2Hz,1H),7.55(dd,J＝8.2,5.2Hz,1H),7.16(t,J＝8.6Hz,1H),5.05(s,2H).
The sixth step
Preparation of 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
4- (bromomethyl) -7-fluoro-1,3-benzothiazole 151f (270mg, 1.09mmol), 4-fluoro-2-methoxy-5-nitroaniline (246mg, 1.31mmol), potassium carbonate (303mg, 2.19mmol) and potassium iodide (91mg, 0.54mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 23%) to give 151g of 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole (270 mg, white solid) in yield: 68 percent.
MS m/z(ESI)：353.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.09(d,J＝1.0Hz,1H),7.85(d,J＝7.2Hz,1H),7.67(dd,J＝8.2,5.2Hz,1H),7.21(t,J＝8.6Hz,1H),6.73(d,J＝12.4Hz,1H),5.71(s,2H),3.95(s,3H).
Seventh step
Preparation of 2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline
Dissolve 151g (150mg, 0.43mmol) of 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole, iron powder (71mg, 1.27mmol) and ammonium chloride (68mg, 1.27mmol) in ethanol: water =1:1 (10 mL), the reaction was stirred at 60 ℃ for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was quenched with water (20 mL), extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline 151h (130 mg, brown solid) in yield: 71 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：323.1[M+1] + 。
The eighth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((7-fluoropheno [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline 151h (130mg, 0.40mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (162mg, 0.48mmol) and triethylamine (81mg, 0.81mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction is finished, the crude product 4- (3- (2-fluoro-5- ((7-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dimethyl dicarboxylate 151i is used in the next step without treatment.
MS m/z(ESI)：564.0[M+1] + 。
The ninth step
Preparation of 3- { 2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolving 4- (3- (2-fluoro-5- ((7-fluoropheno [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 151i in tetrahydrofuran/methanol/water =1:1:1 (6 mL), then lithium hydroxide (7 mg, 0.27mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction was adjusted to pH <7 with 1M HCl, washed with water (10 mL), extracted with ethyl acetate (3X 10 mL), the combined organic layers were washed with brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA) to give 3- { 2-fluoro-5- [ (7-fluoro-1,3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-068 (6.8 mg, white solid), yield: 15 percent.
MS m/z(ESI)：518.0[M+1] + 。
HPLC:99.71％(214nm),99.53％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.57(s,1H),12.01(s,1H),9.51(d,J＝1.4Hz,1H),7.74(dd,J＝8.2,5.4Hz,1H),7.55-7.28(m,3H),7.14(s,1H),5.46(s,2H),3.82(s,3H).
Example 69
Preparation of 3- {7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzooxazol-5-yl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-069)
Figure BDA0003608368340001451
First step of
Preparation of 2-amino-6-methoxy-4-nitrophenol
2-methoxy-4,6-dinitrophenol 155a (1.00g, 4.67mmol) was dissolved in ethanol (20 mL) and then stannous chloride (2.69g, 14.01mmol) was added. The reaction was stirred at 90 ℃ for 2 hours. After LCMS detection of reaction completion, the reaction was cooled to room temperature. The solvent was distilled off under reduced pressure and purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain the product 2-amino-6-methoxy-4-nitrophenol 155b (300 mg, yellow solid) in yield: 31 percent.
MS m/z(ESI)：185.1[M+1] + 。
Second step of
Preparation of 7-methoxy-5-nitro-1,3-benzoxazole
2-amino-6-methoxy-4-nitrophenol 155b (150mg, 0.81mmol) was dissolved in trimethyl orthoformate (10 mL). The reaction was refluxed at 90 ℃ for 2 hours. After completion of the reaction monitored by LCMS, the reaction was allowed to cool to room temperature and the solvent was distilled off under reduced pressure. Purification on silica gel column (petroleum ether/ethyl acetate = 1/1) gave the product 7-methoxy-5-nitro-1,3-benzoxazole 155c (100 mg, yellow solid) in yield: 60 percent.
MS m/z(ESI)：195.1[M+1] + 。
The third step
Preparation of 5-nitro-1,3-benzoxazol-7-ol
7-methoxy-5-nitro-1,3-benzoxazole 155c (120mg, 0.62mmol) was dissolved in DMF (5 mL) followed by the addition of lithium chloride (52mg, 1.24mmol). The reaction was stirred in a microwave reactor at 160 ℃ for 2h. After completion of the reaction, reverse column (water/acetonitrile = 3/7) gave 5-nitro-1,3-benzoxazol-7-ol 155d (80 mg, yellow solid) in yield: 68 percent.
MS m/z(ESI)：181.1[M+1] + 。
The fourth step
Preparation of 7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -5-nitro-1,3-benzoxazole
5-Nitro-1,3-benzoxazol-7-ol 155d (80mg, 0.44mmol) and 2- (chloromethyl) -3,4-difluoro-1-methoxybenzene (94mg, 0.48mmol) were dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (122mg, 0.88mmol) and potassium iodide (36mg, 0.22mmol). The reaction was stirred at 50 ℃ for 4h. After completion of the reaction, water (20 mL) was added, followed by extraction with dichloromethane (3 × 20 mL), drying over anhydrous sodium sulfate, and the residue obtained after concentration under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 4/1) to give 7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -5-nitro-1,3-benzoxazole 155e (50 mg, yellow solid) in 26% yield.
MS m/z(ESI)：337.1[M+1] + 。
The fifth step
Preparation of 7- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzoxazol-5-amine
7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -5-nitro-1,3-benzoxazole 155e (45mg, 0.13mmol) was dissolved in acetonitrile (10 mL) and saturated ammonium chloride (2 mL), followed by addition of iron powder (74mg, 1.34mmol) and reaction stirred at 25 ℃ for 0.5h after completion of the reaction iron powder was filtered off, then extracted with dichloromethane (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 7- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzoxazole-5-amine 155f (40 mg, yellow solid) in 63% yield.
MS m/z(ESI)：307.1[M+1] + 。
The sixth step
Preparation of dimethyl 4- [ ({ - [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzoxazol-5-yl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
7- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzooxazol-5-amine 155f (40mg, 0.13mmol) was dissolved in tetrahydrofuran (5 mL), followed by addition of 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (52mg, 0.15mmol) and triethylamine (39mg, 0.39mmol). The reaction solution was stirred at 25 ℃ for 16 hours after LCMS monitoring the completion of the reaction, the reaction solution was not post-treated to give 155g of stock solution 4- [ ({ [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzooxazol-5-yl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester.
MS m/z(ESI)：548.1[M+1] + 。
Seventh step
Preparation of 3- {7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzooxazol-5-yl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ - [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzooxazol-5-yl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 155g (5 mL stock solution) was dissolved in methanol with lithium hydroxide (5mg, 0.111mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, the reaction mixture was directly concentrated and subjected to preliminary purification by reverse column (water: acetonitrile = 20) to obtain 3- {7- [ ((2,3-difluoro-6-methoxyphenyl) methoxy ] -1,3-benzoxazol-5-yl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-069 (2.20 mg, white solid) after preparative purification, with a yield of 10%.
MS m/z(ESI)：502.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ8.76(s,1H),7.53(dd,J＝19.6,9.6Hz,1H),7.46(d,J＝1.2Hz,1H),7.33(s,1H),7.04(s,1H),6.98(d,J＝9.6Hz,1H),5.29(s,2H),3.81(m,3H).
Example 70
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-070)
Figure BDA0003608368340001471
First step of
Preparation of methyl 2-methylquinoline-8-carboxylate
8-bromo-2-methylquinoline 156a (1400mg, 6.3mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1383mg, 1.9mmol) and triethylamine (1913mg, 18.9mmol) were dissolved in methanol (40 mL), and then the reaction solution was stirred at 70 ℃ for 8 hours under an atmosphere of carbon monoxide (1.5 MPa). After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified on a silica gel column (ethyl acetate/petroleum ether = 20%) to give methyl 2-methylquinoline-8-carboxylate 156b (1300 mg, grey solid) in yield: 92 percent.
MS m/z(ESI)：202.1[M+1] + 。
Second step of
Preparation of (2-methylquinolin-8-yl) methanol
Methyl 2-methylquinoline-8-carboxylate 156b (600mg, 2.7 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) and a solution of lithium aluminium hydride (1.0M in tetrahydrofuran) (3 mL) was added dropwise to the solution at 0 ℃. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give (2-methylquinolin-8-yl) methanol 156c (300 mg, grey solid) in yield: 56 percent.
MS m/z(ESI)：196.1[M+1] + 。
The third step
Preparation of 8- (bromomethyl) -2-methylquinoline
(2-Methylquinolin-8-yl) methanol 156c (300mg, 1.53mmol) was dissolved in dichloromethane (5 mL) and phosphorus tribromide (0.5 mL) was added dropwise to the solution at 0 ℃. The reaction solution was stirred at 25 ℃ for 2 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 8- (bromomethyl) -2-methylquinoline 156d (180 mg, yellow solid) in yield: 46 percent.
MS m/z(ESI)：236.0[M+1] + . The fourth step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methylquinoline
8- (bromomethyl) -2-methylquinoline 156d (100mg, 0.39mmol), 4-fluoro-2-methoxy-5-nitroaniline (250mg, 1.40mmol), potassium carbonate (1011mg, 5.4 mmol) and potassium iodide (52mg, 0.21mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 50%) to give 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methylquinoline 156e (55 mg, yellow solid) in yield: 40 percent.
MS m/z(ESI)：343.1[M+1] + 。
The fifth step
Preparation of 2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) aniline
8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methylquinoline 156e (55mg, 0.15mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (650mg, 9.4 mmol) and a saturated ammonium chloride solution (5 mL) to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) aniline 156f (50 mg, yellow solid), yield: 96 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.1[M+1] + 。
The sixth step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate
2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) aniline 156f (50mg, 0.13mmol) was dissolved in tetrahydrofuran (6 mL), followed by the addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (686mg, 2.0 mmol) and triethylamine (162mg, 1.52mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 156g (65 mg, yellow solid) of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate, yield: 82 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：554.1[M+1] + 。
Seventh step
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
156g (65mg, 0.11mmol) of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate was dissolved in tetrahydrofuran/water =1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative purification (FA) to give 3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-070 (12 mg, yellow solid), yield: 20 percent.
MS m/z(ESI)：530.0[M+1] + 。
HPLC:97.59％(214nm),97.57％(254nm).
1 H NMR(400MHz,DMSO-d6)δ11.77(s,1H),8.29(d,J＝8.4Hz,1H),7.93-7.87(m,2H),7.61-7.55(m,1H),7.46(d,J＝8.4Hz,1H),7.34(d,J＝6.6Hz,1H),7.14(d,J＝11.4Hz,2H),5.62(s,2H),3.85(s,3H),2.64(s,3H)
Example 71
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-071)
Figure BDA0003608368340001481
First step of
Preparation of 8-bromo-3-methylquinoline
2-bromoaniline 157a (10g, 5.8mmol) and 2-methylprop-2-enal (6.12g, 8.8mmol) were dissolved in 6N hydrochloric acid (10 mL). The reaction was stirred at 110 ℃ for 16 h under nitrogen. After the reaction was complete, the reaction was quenched with 5N sodium hydroxide solution to pH >9, extracted with dichloromethane (3X 20 mL) and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by silica gel column (ethyl acetate/petroleum ether = 7%) to give 8-bromo-3-methylquinoline 157b (2.5 g, yellow liquid), yield: 19 percent.
MS m/z(ESI)：222.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.92(d,J＝2.0Hz,1H),8.10-7.86(m,2H),7.73(dd,J＝8.2,1.0Hz,1H),7.38(t,J＝7.8Hz,1H),2.56(s,3H).
Second step of
Preparation of methyl 3-methylquinoline-8-carboxylate
8-bromo-3-methylquinoline 157b (1.5g, 6.8mmol) and 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride (2.49g, 3.4 mmol) were dissolved in methanol (10 mL), triethylamine (1.38g, 13.6 mmol) was added at room temperature, and then the reaction solution was stirred at 100 ℃ for 8 hours under an atmosphere of carbon monoxide (1.5 MPa). After completion of the reaction, the reaction solution was filtered, water (20 mL) was added to the filtrate, extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (ethyl acetate/petroleum ether = 30%) to give methyl 3-methylquinoline-8-carboxylate 157c (270 mg, yellow solid), yield: 20 percent.
MS m/z(ESI)：202.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.93(d,J＝2.0Hz,1H),8.03-7.92(m,2H),7.88(dd,J＝8.2,1.2Hz,1H),7.59-7.48(m,1H),4.06(s,3H),2.54(s,3H).
The third step
Preparation of (3-methylquinolin-8-yl) methanol
Methyl 3-methylquinoline-8-carboxylate 157c (100mg, 0.50mmol) was dissolved in tetrahydrofuran (10 mL) and diisobutyl lithium hydride (0.5mL, 0.50mmol) was added dropwise at-78 ℃. The reaction solution was stirred at-78 ℃ for 1 hour. After the reaction was completed, the reaction solution was quenched with saturated sodium potassium tartrate solution (10 mL), washed with water (20 mL), extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give crude (3-methylquinolin-8-yl) methanol 157d (90 mg, yellow solid) as a crude product, yield: 94 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：174.3[M+1] + 。
The fourth step
Preparation of 8- (bromomethyl) -3-methylquinoline
(3-Methylquinolin-8-yl) methanol 157d (70mg, 0.40mmol) was dissolved in dichloromethane (5 mL) and phosphorus tribromide (33mg, 0.12mmol) was added dropwise to the solution at 0 ℃. The mixture was stirred at 0 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (10 mL), extracted with dichloromethane (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration gave the crude 8- (bromomethyl) -3-methylquinoline 157e (73 mg, yellow solid) as a yield: 72 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：238.1(M+H)。
The fifth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline
8- (bromomethyl) -3-methylquinoline 157e (73mg, 0.31mmol)), 4-fluoro-2-methoxy-5-nitroaniline (69mg, 0.37mmol), potassium carbonate (85mg, 0.62mmol) and potassium iodide (25mg, 0.15mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 23%) to give 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline 157f (60 mg, white solid) in yield: 56 percent.
MS m/z(ESI)：343.1[M+1] + 。
The sixth step
Preparation of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline
Dissolve 78- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline 157f (50mg, 0.15mmol), iron powder (24mg, 0.44mmol) and ammonium chloride (23mg, 0.44mmol) in ethanol: water =1:1 (5 mL), and the reaction was stirred at 60 ℃ for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was quenched with water (10 mL), extracted with ethyl acetate (3X 10 mL), and the combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 157g (60 mg, brown solid) of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline (yield: 86 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.3[M+1] + 。
Step seven
Preparation of 4- [ ({ { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester
157g (65mg, 0.21mmol) of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline was dissolved in tetrahydrofuran (5 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (83mg, 0.24mmol) and triethylamine (42mg, 0.42mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, filtration and concentration gave 4- [ ({ { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 157h (30 mg, yellow solid) in yield: 25 percent. The crude product was used directly in the next reaction
MS m/z(ESI)：554.0[M+1] + 。
The eighth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 157h (30mg, 0.05mmol) was dissolved in tetrahydrofuran/water =1:1 (6 mL), then lithium hydroxide (4 mg, 0.16mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was adjusted to pH <7 with 1M HCl, washed with water (10 mL), extracted with ethyl acetate (3X 10 mL), the combined organic layers were washed with brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA) to give 3- { 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-071 (9.71 mg, white solid) yield: 36 percent.
MS m/z(ESI)：508.1[M+1] + 。
HPLC:99.01％(214nm),96.63％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.95(s,1H),8.79(d,J＝2.4Hz,1H),8.18(d,J＝0.8Hz,1H),7.88(dd,J＝14.8,7.6Hz,2H),7.65–7.60(m,1H),7.39(d,J＝7.4Hz,1H),7.34(s,1H),7.15(d,J＝11.6Hz,1H),5.61(s,2H),3.84(s,3H),2.51(s,3H).
Example 72
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (pd-072)
Figure BDA0003608368340001501
First step of
Preparation of 2-methoxy-8-methylquinoline
Compound 158a (500mg, 2.81mmol) and CH 3 ONa (1.52g, 28.1mmol) in CH 3 OH (10 mL) and reacted at 65 ℃ for 48h. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in water (10 mL), extracted with ethyl acetate (3X 10 mL), washed with saturated NaCl, and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE) to give 2-methoxy-8-methylquinoline 158b (250 mg, colorless oily liquid), yield: 49 percent.
MS m/z(ESI)：174.0[M+1] + .
Second step of
Preparation of 8- (bromomethyl) -2-methoxyquinoline
Compound 158b (100mg, 0.58mmol), BPO (7mg, 0.03mmol) and NBS (113mg, 0.64mmol) were dissolved in CCl 4 (10 mL) at 80 ℃ for 2h. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (PE) to give 8- (bromomethyl) -2-methoxyquinoline 158c (120 mg, white solid), yield: 83 percent.
MS m/z(ESI)：252.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ7.99(d,J＝8.8Hz,1H),7.73(dd,J＝7.2,1.4Hz,1H),7.70(dd,J＝8.0,1.4Hz,1H),7.34(dd,J＝8.0,7.2Hz,1H),6.95(d,J＝8.8Hz,1H),5.15(s,2H),4.13(s,3H).
The third step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methoxyquinoline
The compound 158c (100mg, 0.40mmol), 4-fluoro-2-methoxy-5-naphthol (82mg, 0.44mmol) and K 2 CO 3 (165mg, 1.2mmol) was dissolved in ACN (10 mL) and reacted at 80 ℃ for 2 hours. The reaction was filtered, the filtrate was concentrated, and the resulting residue was purified by column chromatography (EtOAc: PE = 35%) to give 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methoxyquinoline 158d (100 mg, white solid), yield: 70 percent.
MS m/z(ESI)：359.1[M+1] + .
The fourth step
Preparation of 2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) aniline
Compound 158d (90mg, 0.25mmol) is dissolved in MeOH NH 4 To a solution of Cl =5:1 (12 mL), fe powder (140mg, 2.5 mmol) was added and reacted at room temperature for 2h. The reaction solution was filtered, the filtrate was concentrated, the obtained residue was dissolved in water, extracted with ethyl acetate, washed with saturated NaCl water, and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) aniline 158e (80 mg, light brown solid) in yield: 96 percent.
MS m/z(ESI)：329.2[M+1] + .
The fifth step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate
Compound 158e (70mg, 0.21mmol) and TEA (65mg, 0.63mmol) were dissolved in anhydrous THF (5 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (107mg, 0.31mmol) and reacted at room temperature for 16h. The reaction was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (EtOAc: PE = 30%) to give 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 158d (60 mg, yellow solid), yield: 45 percent.
MS m/z(ESI)：570.1[M+1] + .
The sixth step
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 158d (60mg, 0.11mmol) was dissolved in THF/MeOH/H 2 O = 3. The reaction mixture was concentrated under reduced pressure, and the residue obtained was subjected to preparative HPLC (ACN: H) 2 O (0.1% by weight FA) = 72%) was purified to give 3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d]Pyrimidine-5-carboxylic acid Cpd-072 (27.32 mg, white solid), yield: and 48 percent.
MS m/z(ESI)：524.1[M+1] + .
1 H NMR(400MHz,DMSO-d 6 )δ14.55(s,1H),11.96(s,1H),8.27(d,J＝8.8Hz,1H),7.91-7.85(m,1H),7.82(d,J＝7.2Hz,1H),7.51-7.45(m,1H),7.36(s,1H),7.34(d,J＝7.4Hz,1H),7.15(d,J＝11.6Hz,1H),7.04(d,J＝8.8Hz,1H),5.58(s,2H),3.91(s,3H),3.85(s,3H).
19 F NMR(376MHz,DMSO-d 6 )δ-128.59(s,1H).
Example 73
Preparation of 3- (2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-073)
Figure BDA0003608368340001521
First step of
Preparation of quinoline-8-carboxylic acid methyl ester
Compound 159a (1 g,5.8 mmol) is dissolved in dry DCM (25 mL), oxalyl chloride (1.1 g,8.7 mmol) and a catalytic amount of DMF are added under ice bath conditions, and the reaction is carried out at 0 ℃ for 1h followed by MeOH (10 mL). The reaction was concentrated under reduced pressure and the residue was dissolved in EtOAc (20 mL) and separately washed with saturated NaHCO 3 And saturated NaCl water washing, dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 50%) to give quinoline-8-carboxylic acid methyl ester 159b (0.9 g, orange yellow oily liquid), yield: 79 percent.
MS m/z(ESI)：188.0[M+1] + .
Second step of
Preparation of methyl 3-hydroxyquinoline-8-carboxylate
Compound 159b (900mg, 4.8mmol) was dissolved in AcOH (5 mL), followed by addition of H 2 O 2 (30%, 1 mL) and reacted at 85 ℃ for 4h. Pouring the reaction solution into NaHCO 3 In the solution, DCM is extracted, saturated NaCl solution is washed with water, and anhydrous sodium sulfate is dried. Filtration and concentration of the filtrate under reduced pressure gave a residue which was purified by column chromatography (MeOH: DCM = 5%) to give methyl 3-hydroxyquinoline-8-carboxylate 159c (200 mg, light brown solid) in yield: 21 percent. MS m/z (ESI): 204.2[ M ] +1 ] + .
The third step
Preparation of methyl 3-methoxyquinoline-8-carboxylate
Compound 159c (200mg, 1mmol), K 2 CO 3 (408mg, 3mmol) and CH 3 I (210mg, 1.5 mmol) was dissolved in anhydrous DMF (10 mL) and reacted at 80 ℃ for 4h. The reaction was poured into water, extracted with EtOAc, washed with saturated NaCl, and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure the resulting residue was purified by column chromatography (EtOAc: PE = 30%) to give methyl 3-methoxyquinoline-8-carboxylate 159d (200 mg, orange solid) in yield: 89 percent.
MS m/z(ESI)：218.2[M+1] + .
The fourth step
Preparation of (3-methoxyquinolin-8-yl) methanol
Compound 159d (250mg, 0.92mmol) was dissolved in anhydrous THF (10 mL), and LiAlH4 (218mg, 5.75mmol) was added at 0 ℃ and reacted at 0 ℃ for 20min. The reaction solution is saturated with NH 4 The Cl solution was quenched, extracted with EtOAc, washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure the resulting residue was purified by column chromatography (EtOAc: PE = 35%) to give (3-methoxyquinolin-8-yl) methanol 159e (125 mg, yellow oily liquid), yield: and 55 percent.
MS m/z(ESI)：190.2[M+1] + .
The fifth step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -3-methoxyquinoline
Compound 159e (125mg, 0.66mmol), 4-fluoro-2-methoxy-5-nitroPhenylphenol (136mg, 0.73mmol) and PPh 3 (225mg, 0.86mmol) was dissolved in dry THF (10 mL) and DIAD (174mg, 0.86mmol) was added dropwise and reacted at 45 ℃ for 2h. Cooled to room temperature and filtered to give 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -3-methoxyquinoline 159f (100 mg, light yellow solid), yield: 40 percent.
MS m/z(ESI)：359.1[M+1] + .
The sixth step
Preparation of 2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) aniline
Compound 159f (90mg, 0.25mmol) was dissolved in MeOH (10 mL) and saturated NH 4 To a mixed solution of Cl (2 mL), iron powder (140mg, 2.5mmol) was then added. Reacting at room temperature for 2h, filtering, and concentrating the filtrate under reduced pressure. The resulting residue was dissolved in ethyl acetate (20 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 159g (70 mg, light brown solid) of 2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) aniline in yield: 68 percent.
MS m/z(ESI)：329.1[M+1] + .
Seventh step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylate
159g (70mg, 0.21mmol) of the compound and TEA (65mg, 0.64mmol) were dissolved in anhydrous THF (5 mL), and then 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (107mg, 0.32mmol) was added and reacted at room temperature for 16h. The reaction was concentrated under reduced pressure to give 4- (3- (2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 159h (60 mg, yellow solid), yield: 40 percent.
MS m/z(ESI)：570.1[M+1] + .
Eighth step
Preparation of 3- (2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 159H (60mg, 0.11mmol) was dissolved in THF/MeOH/H 2 O =3mmol), and reacted at room temperature for 1h. The reaction mixture was concentrated under reduced pressure, and the residue obtained was subjected to preparative HPLC (ACN: H) 2 O(0.1％NH 3 ) = 90%) to obtain 3- (2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) phenyl) -2,4-dioxy-1H-thieno [3,4-d]Pyrimidine-5-carboxylic acid Cpd-073 (0.71 mg, white solid), yield: 1.3 percent.
MS m/z(ESI)：524.1[M+1] + .
1 H NMR(400MHz,CD 3 OD)δ8.51(s,1H),7.76(dd,J＝13.0,7.8Hz,2H),7.63(s,1H),7.58-7.49(m,1H),7.10(d,J＝5.8Hz,1H),6.94(d,J＝10.6Hz,2H),5.65(s,2H),3.94(s,3H),3.89(s,3H).
Example 74
Preparation of 3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-074)
Figure BDA0003608368340001541
First step of
Preparation of 3- (bromomethyl) benzothiophene
3-methylbenzothiophene 160a (200mg, 1.35mmol), N-bromosuccinimide (288mg, 1.62mmol) and azobisisobutyronitrile (110mg, 0.67mmol) were dissolved in carbon tetrachloride (5 mL), followed by stirring for 2 hours at 85 ℃ under a nitrogen atmosphere. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 40 mL), and the combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained crude product was purified by silica gel column (petroleum ether) to obtain 3- (bromomethyl) benzothiophene 160b (100 mg, white solid) in yield: 32 percent.
1 H NMR(400MHz,CDCl 3 )δ7.93-7.83(m,2H),7.50(s,1H),7.49-7.43(m,1H),7.43-7.36(m,1H),4.76(s,2H).
Second step of
Preparation of 3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene
3- (bromomethyl) benzothiophene 160b (100mg, 0.39mmol), 4-fluoro-2-methoxy-5-nitrophenol (250mg, 1.40mmol), potassium carbonate (1011mg, 5.4 mmol), and potassium iodide (52mg, 0.21mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 50%) to give 3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene 160c (55 mg, white solid), yield: 40 percent.
MS m/z(ESI)：356.1[M+1] + 。
The third step
Preparation of 5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyaniline
3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene 160c (55mg, 0.15mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (650mg, 9.4 mmol) and saturated ammonium chloride solution (5 mL) to the solution. The reaction was stirred at room temperature for 2 hours under nitrogen. After the reaction was completed, the reaction solution was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyaniline 160d (50 mg, yellow solid), yield: 96 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：304.1[M+1] + 。
The fourth step
Preparation of 4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- (Benzothien-3-ylmethoxy) -2-fluoro-4-methoxyaniline 160d (50mg, 0.13mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (686mg, 2.0mmol) and triethylamine (162mg, 1.52mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 160e (65 mg, yellow solid), yield: 82 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：545[M+1] + 。
The fifth step
Preparation of 3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 160e (65mg, 0.11mmol) was dissolved in tetrahydrofuran/water =1:1 (5 mL), then lithium hydroxide (108mg, 2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by preparative (NH) 3 ) To obtain 3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d]Pyrimidine-5-carboxylic acid Cpd-074 (10 mg, yellow solid), yield: 20 percent.
MS m/z(ESI)：408.9[M+1] + 。
HPLC:99.63％(214nm),98.72％(254nm).
1 H NMR(400MHz,DMSO)δ11.64(s,1H),8.02(d,J＝6.4Hz,1H),7.92(d,J＝6.8Hz,1H),7.87(s,1H),7.42(s,2H),7.33(d,J＝7.2Hz,1H),7.12(d,J＝11.6Hz,1H),7.03(s,1H),5.25(s,2H),3.81(s,3H).
Example 75 and example 76
Preparation of 3- { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-075) and 3- (2-fluoro-5- ((6-fluorobenzo [ d ] thiazol-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-076)
Figure BDA0003608368340001551
First step of
Preparation of 1-benzoyl-3- (4-fluoro-3-methylphenyl) thiourea
4-fluoro-3-methylaniline 163a (3 g, 24mmol) was dissolved in tetrahydrofuran (10 mL). Benzoyl isothiocyanate (7.83g, 48mmol) was then added at 25 ℃. The reaction mixture was reacted at 25 ℃ for 16 hours. Then spin-drying the solvent and then mixing with ethanol: water =4:1 (50 mL) was filtered and the residue was collected to give 1-benzoyl-3- (4-fluoro-3-methylphenyl) thiourea 163b (7 g, white solid) in 90% yield.
MS m/z(ESI)：289[M+1] + 。
Second step of
Preparation of (4-fluoro-3-methylphenyl) thiourea
1-benzoyl-3- (4-fluoro-3-methylphenyl) thiourea 163b (8.00g, 27.7 mmol) was dissolved in methanol: water =3:1 (60 mL). Potassium carbonate (11.49g, 83.1 mmol) was then added at 25 ℃. The reaction mixture was reacted at 70 ℃ for 2 hours. Then, the solvent was spin-dried, and water was added to filter the residue to give (4-fluoro-3-methylphenyl) thiourea 163c (4 g, white solid) in a yield of 79%.
MS m/z(ESI)：185[M+1] + 。
The third step
Preparation of 6-fluoro-7-methyl-1,3-benzothiazol-2-amine
(4-fluoro-3-methylphenyl) thiourea 163c (4.00g, 21mmol) was dissolved in chloroform (50 mL) and liquid bromine (4.16g, 23mmol) was added dropwise at 25 ℃. The reaction mixture was reacted at 60 ℃ for 2 hours. Then the solvent is dried by spinning, water is added, and filter residue is filtered to obtain a mixture of 6-fluoro-7-methyl-1,3-benzothiazole-2-amine 163d and 6-fluoro-5-methylbenzo [ d ] thiazole-2-amine 170d (2g, 170d =2, white solid), the yield is 45%, the mixture cannot be separated by a column, the peak time of high performance liquid chromatography is basically the same, the next steps are put together for reaction and treatment, and the synthesis steps of the molecule Cpd-076 from the fourth step to the ninth step are not specially described, taking the synthesis step of Cpd-075 as an example, and can not be separated until the final step is prepared.
MS m/z(ESI)：183[M+1] + 。
The fourth step
Preparation of 6-fluoro-7-methyl-1,3-benzothiazole
6-fluoro-7-methyl-1,3-benzothiazol-2-amine 163d (1.8g, 0.01mol) was dissolved in 1,4-dioxane (20 mL). Thereafter, tert-butyl nitrite (3.06g, 0.30mol) was added to the reaction mixture, and the reaction mixture was reacted at 90 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 60/40) to give 6-fluoro-7-methyl-1,3-benzothiazole 163e (0.8 g, brown solid) in 43% yield.
MS m/z(ESI)：168[M+1] + 。
The fifth step
Preparation of 5- (bromomethyl) -6,8-difluoroquinoxaline
6-fluoro-7-methyl-1,3-benzothiazole 163e (650mg, 3.6mmol) was dissolved in carbon tetrachloride (15 mL). Then dibenzoyl peroxide (436.mg, 1.8mmol) and N-bromosuccinimide [706mg,3.96mmol ] were added at 25 ℃ and the reaction solution was reacted at 85 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 5- (bromomethyl) -6,8-difluoroquinoxaline 163f (500 mg, red solid) in 48% yield.
MS m/z(ESI)：259.0[M+1] + 。
The sixth step
Preparation of 6-fluoro-7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
5- (bromomethyl) -6,8-difluoroquinoxaline 163f (600mg, 2.44mmol) and 4-fluoro-2-methoxy-5-nitrophenol (456 mg, 2.43mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (673mg, 4.87mmol) and potassium iodide (607mg, 3.66mmol) were added, and the reaction mixture was reacted at 65 ℃ for 2 hours. The solvent was then spun dry and extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 163g (500 mg, yellow solid) of 6-fluoro-7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole in 52% yield.
MS m/z(ESI)：353.0[M+1] + 。
Seventh step
Preparation of 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyaniline
163g (500mg, 1.4 mmol) of 6-fluoro-7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (79mg, 1.4 mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyaniline 163h (400 mg, brown solid) in 79% yield.
MS m/z(ESI)：323.0[M+1] + 。
Eighth step
Preparation of dimethyl 4- [ ({ { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyaniline 163h (450mg, 1.4 mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (468mg, 1.39mmol) were dissolved in tetrahydrofuran (10 mL), then triethylamine (423mg, 4.20mmol) was added, and the reaction liquid was reacted at 25 ℃ for 1 hour to obtain a stock solution 4- [ ({ { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 163i without any post-treatment.
MS m/z(ESI)：564.0[M+1] + 。
The ninth step
Preparation of 3- { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 163i (10 mL stock solution) was dissolved in methanol with lithium hydroxide (21mg, 0.078mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was completed, 3- { 2-fluoro-5- [ (6-fluoro-1,3-benzothiazol-7-yl) methoxy ] -4-methoxyphenyl } -2,4-dioxo-1H thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-075 (2.84 mg, white solid) was obtained after direct concentration and preparative purification by reverse column (water: acetonitrile = 5).
MS m/z(ESI)：518.0[M+1] + 。
HPLC:96.58％(214nm),95.15％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.53(s,1H),12.01(s,1H),9.42(s,1H),8.14(dd,J＝8.8,4.4Hz,1H),7.49(dd,J＝9.6,9.2Hz,1H),7.39(s,1H),7.32(d,J＝7.2Hz,1H),7.20(d,J＝11.6Hz,1H),5.32(s,2H),3.86(s,3H).
3- (2-fluoro-5- ((6-fluorobenzo [ d ] thiazol-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxy-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-076 (6.21 mg, white solid) yield: 32%.
MS m/z(ESI)：518.0[M+1] + 。
HPLC:95.18％(214nm),95.00％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.57(s,1H),12.04(s,1H),9.41(d,J＝6.0Hz,1H),8.28(d,J＝6.4Hz,1H),8.15(d,J＝9.6Hz,1H),7.40(d,J＝2.4Hz,1H),7.37(d,J＝7.2Hz,1H),7.17(d,J＝11.6Hz,1H),5.18(s,2H),3.96(s,3H).
Example 77 and example 78
Preparation of 3- [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-077) and 3- [5- (1,3-benzothiazol-5-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-078)
Figure BDA0003608368340001571
First step of
Preparation of 1-benzoyl-3- (3-methylphenyl) thiourea
3-methylaniline 165a (5g, 0.05mol) was dissolved in tetrahydrofuran (80 mL). Benzoyl isothiocyanate (15g, 0.09mol) was then added at 25 ℃. The reaction mixture was reacted at 25 ℃ for 16 hours. Then spin-drying the solvent and then mixing with ethanol: water =4:1 (50 mL) was filtered to obtain the residue which was used to give 1-benzoyl-3- (3-methylphenyl) thiourea 165b (14 g, yellow solid) in 88% yield.
MS m/z(ESI)：271.1[M+1] + 。
Second step of
Preparation of (3-methylphenyl) thiourea
Dissolve 1-benzoyl-3- (3-methylphenyl) thiourea 165b (5g, 0.02mol) in methanol: water =3:1 (60 mL). Potassium carbonate (7.67g, 0.05mol) was then added at 25 ℃. The reaction mixture was reacted at 70 ℃ for 2 hours. Then, the solvent was spin-dried, water was added, and filtration was carried out to obtain the residue as (3-methylphenyl) thiourea 165c (2.5 g, white solid) in a yield of 64%.
MS m/z(ESI)：167.1[M+1] + 。
The third step
Preparation of 7-methyl-1,3-benzothiazol-2-amine
(3-methylphenyl) thiourea 165c (1.8g, 0.01mol) was dissolved in chloroform (50 mL) and then liquid bromine (2.1g, 0.01mol) was added dropwise at 25 ℃. The reaction mixture was reacted at 60 ℃ for 2 hours. Then the solvent is dried by spinning, water is added, filtration is carried out, and filter residues are obtained to obtain a mixture of 7-methyl-1,3-benzothiazole-2-amine 165d and 5-methyl-1,3-benzothiazole-2-amine 171d (1g, 171d =1, white solid), the yield is 45%, the mixture cannot be separated by a column, the peak time of high performance liquid chromatography is basically the same, the next steps are put together for reaction and treatment, the synthesis steps of the molecule Cpd-078 from the fourth step to the ninth step are not specially explained, and the synthesis steps of Cpd-077 are taken as an example, and can not be separated until the last step is prepared.
MS m/z(ESI)：165.1[M+1] + 。
The fourth step
Preparation of 7-methyl-1,3-benzothiazole
7-methyl-1,3-benzothiazol-2-amine 165d (1g, 0.006mol) was dissolved in 1,4-dioxane (20 mL). Thereafter, tert-butyl nitrite (1.9g, 0.018mol) was added thereto, and the reaction solution was reacted at 90 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 60/40) to give 7-methyl-1,3-benzothiazole 165e (0.5 g, brown solid) in 44% yield.
MS m/z(ESI)：150.1[M+1] + 。
The fifth step
Preparation of 7- (bromomethyl) -1,3-benzothiazole
7-methyl-1,3-benzothiazole 165e (600mg, 4.02mmol) was dissolved in carbon tetrachloride (15 mL). NBS (858 mg, 4.82mmol) and BPO (487mg, 2.01mmol) were then added thereto at 25 ℃ and the reaction mixture was reacted at 85 ℃ for 2 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 7- (bromomethyl) -1,3-benzothiazole 165f (500 mg, yellow solid) in 43% yield.
MS m/z(ESI)：228.1[M+1] + 。
The sixth step
Preparation of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
7- (bromomethyl) -1,3-benzothiazole 165f (800mg, 3.50mmol) and 4-fluoro-2-methoxy-5-nitrophenol (787 mg, 4.20mmol) were dissolved in acetonitrile (30 mL). Then, potassium carbonate (969mg, 7.01mmol) and potassium iodide (873mg, 5.26mmol) were added, and the reaction solution was reacted at 65 ℃ for 2 hours. The solvent was then spun dry and extracted with dichloromethane (2X 50 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 165g of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole (600 mg, yellow solid) in 40% yield.
MS m/z(ESI)：335.0[M+1] + 。
Seventh step
Preparation of 5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyaniline
7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 165g (300mg, 0.89mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (1g, 17mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2X 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyaniline 165h (250 mg, brown solid) in 73% yield.
MS m/z(ESI)：305.1[M+1] + 。
The eighth step
Preparation of dimethyl 4- ({ [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate
5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyaniline 165h (200mg, 0.65mmol) and 4- [ ((phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (550mg, 1.64mmol) were dissolved in tetrahydrofuran (10 mL), and then triethylamine (368mg, 6.57mmol) was added, and the reaction solution was reacted at 25 ℃ for 16 hours to obtain a stock solution 4- ({ [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 165i without any post-treatment.
MS m/z(ESI)：564.0[M+1] + 。
The ninth step
Preparation of 3- [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- ({ [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylate 165i (10 mL stock solution) was dissolved in methanol with lithium hydroxide (61mg, 1.28mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 90 minutes. After the reaction was finished, 3- [5- (1,3-benzothiazol-7-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-077 (27.03 mg, white solid) was obtained after direct concentration, preliminary purification by reverse column (water: acetonitrile = 5).
MS m/z(ESI)：500.1[M+1] + 。
HPLC:100％(214nm),99.43％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.52(s,1H),11.98(s,1H),9.44(s,1H),8.10(dd,J＝7.2,2.0Hz,1H),7.60-7.55(m,2H),7.37(s,1H),7.33(d,J＝7.2Hz,1H),7.18(d,J＝11.6Hz,1H),5.29(s,2H),3.86(s,3H).
3- [5- (1,3-benzothiazol-5-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-078 (88.86 mg, white solid) yield 65%.
MS m/z(ESI)：500.0[M+1] + 。
HPLC:98.40％(214nm),95.02％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.31(s,1H),9.43(s,1H),8.20-8.17(m,2H),7.57(dd,J＝8.4,1.2Hz,1H),7.21(d,J＝7.2Hz,1H),7.10(d,J＝11.6Hz,1H),6.62(s,1H),5.16(d,J＝2.4Hz,2H),3.84(s,3H).
Example 79
Preparation of 3- {5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-079)
Figure BDA0003608368340001601
First step of
Preparation of 4-chloro-2-fluoro-6-nitroaniline
2-fluoro-6-nitroaniline 166a (2g, 12.8 mmol) and N-chlorosuccinimide (1.71g, 12.8 mmol) were dissolved in N, N-dimethylformamide (20 mL). The reaction solution was stirred at 80 ℃ for 16 hours under nitrogen. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 8%) to give 4-chloro-2-fluoro-6-nitroaniline 166b (2.13 g, yellow solid), yield: 86 percent.
MS m/z(ESI)：191.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ7.96(t,J＝2.4Hz,1H),7.25(dd,J＝9.6,3.2Hz,1H),6.12(s,2H).
Second step of
Preparation of 2-bromo-5-chloro-1-fluoro-3-nitrobenzene
Cuprous bromide (2.82g, 19.6 mmol) and tert-butyl nitrite (4.05g, 39.3 mmol) were dissolved in acetonitrile (10 mL), and a solution of 4-chloro-2-fluoro-6-nitroaniline 166b (2.5g, 13.1mmol) in acetonitrile (10 mL) was added dropwise at 50 ℃. The reaction solution was stirred at 50 ℃ for 2 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 5%) to give 2-bromo-5-chloro-1-fluoro-3-nitrobenzene 166c (2.5 g, yellow solid), yield: 73 percent.
1 H NMR(400MHz,CDCl 3 )δ7.68(t,J＝2.0Hz,1H),7.40(dd,J＝7.6,2.4Hz,1H).
The third step
Preparation of 2-bromo-5-chloro-3-fluoroaniline
2-bromo-5-chloro-1-fluoro-3-nitrobenzene 166c (2.5g, 9.8mmol), iron powder (1.64g, 29.4mmol) and ammonium chloride (1.57g, 29.4mmol) were dissolved in ethanol: water =1:1 (30 mL), and the reaction mixture was stirred at 60 ℃ for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was quenched with water (10 mL), extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 2-bromo-5-chloro-3-fluoroaniline 166d (1.8 g, brown solid) in yield: 77 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：223.9[M+1] + 。
The fourth step
Preparation of 1-benzoyl-3- (2-bromo-5-chloro-3-fluorophenyl) thiourea
2-bromo-5-chloro-3-fluoroaniline 166d (2g, 8.9mmol) was dissolved in tetrahydrofuran (30 mL) and benzoyl isothiocyanate (2.9g, 17.8mmol) was added dropwise to the solution. The reaction solution was stirred at 25 ℃ for 16 hours under air protection. After the reaction was complete, the solvent was removed in vacuo and the solid was washed with a mixed solvent of ethanol: petroleum ether =4:1 (50 mL) and filtered. The filter cake was 1-benzoyl-3- (2-bromo-5-chloro-3-fluorophenyl) thiourea 166e (3.4 g, white solid), yield: 96 percent.
MS m/z(ESI)：387.0(M+H)。
The fifth step
Preparation of (2-bromo-5-chloro-3-fluorophenyl) thiourea
1-benzoyl-3- (2-bromo-5-chloro-3-fluorophenyl) thiourea 166e (3.4g, 8.8mmol) and potassium carbonate (2.43g, 17.6mmol) were dissolved in methanol water =4:1 (20 mL), and the reaction was stirred at 70 ℃ for 2 hours. After the reaction was complete, the solvent was removed in vacuo and then water (30 mL) was added. The solid was collected by filtration and further washed with water (20 mL). Filter cake was (2-bromo-5-chloro-3-fluorophenyl) thiourea 166f (1.58 g, white solid), yield: 62 percent. MS m/z (ESI): 283.0[ M ] +1] + 。
The sixth step
Preparation of 4-bromo-7-chloro-5-fluoro-1,3-benzothiazol-2-amine
(2-bromo-5-chloro-3-fluorophenyl) thiourea 166f (1.58g, 5.6 mmol) was dissolved in chloroform (20 mL), and liquid bromine (1.07g, 6.7 mmol) was added dropwise at room temperature. The reaction solution was stirred at 70 ℃ for 18 hours. After the reaction was completed, the solvent was removed in vacuo, and the solid was dissolved in a mixed solvent of dichloromethane: methanol = 1: 1 (20 mL), to which an aqueous solution of sodium thiosulfate (50 mL) was added. The solvent was then removed in vacuo and water (50 mL) was added. The solid was collected by filtration and the filter cake was 166g (1.8 g, yellow solid) of 4-bromo-7-chloro-5-fluoro-1,3-benzothiazol-2-amine, yield: 94 percent.
MS m/z(ESI)：281.0[M+1] + 。
Seventh step
Preparation of 4-bromo-7-chloro-5-fluoro-1,3-benzothiazole
166g (1.8g, 6.4 mmol) of 4-bromo-7-chloro-5-fluoro-1,3-benzothiazol-2-amine was dissolved in 1,4-dioxane (20 mL), and tert-butyl nitrite (1.98g, 19.2 mmol) was added dropwise to the solution at room temperature. The mixture was stirred at 90 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (10 mL), extracted with ethyl acetate (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 6%) to give 4-bromo-7-chloro-5-fluoro-1,3-benzothiazole 166h (1.05 g, yellow solid), yield: 61 percent.
MS m/z(ESI)：265.9[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.65(s,1H),7.91(d,J＝8.8Hz,1H).
Eighth step
Preparation of 7-chloro-5-fluoro-1,3-benzothiazole-4-methyl formate
4-bromo-7-chloro-5-fluoro-1,3-benzothiazole 166h (1.05g, 3.9mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1.43g, 1.97mmol) and triethylamine (954mg, 7.88mmol) were dissolved in methanol (20 mL) and the reaction solution was subsequently stirred at 100 ℃ for 24 h under an atmosphere of carbon monoxide (1.5 MPa). After completion of the reaction, the reaction solution was filtered, water (20 mL) was added to the filtrate, extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 15%) to give 7-chloro-5-fluoro-1,3-benzothiazole-4-carboxylic acid methyl ester 166i (174 mg, white solid) in yield: 18 percent.
MS m/z(ESI)：246.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ9.21(s,1H),7.38(d,J＝9.2Hz,1H),4.09(s,3H).
The ninth step
Preparation of (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methanol
7-chloro-5-fluoro-1,3-benzothiazole-4-carboxylic acid methyl ester 166i (150mg, 0.61mmol) is dissolved in anhydrous tetrahydrofuran (10 mL) and a diisobutylaluminum hydride solution (1.0M in hexanes) (1.2 mL) is added dropwise to the solution at 0 ℃. The mixture was stirred at 0 ℃ for 2 hours. After the reaction was complete, the reaction was quenched with saturated sodium potassium tartrate (10 mL), washed with water (10 mL), extracted with ethyl acetate (3 × 20 mL), and the combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methanol 166j (130 mg, yellow solid), yield: 88 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：218.0[M+1] + 。
The tenth step
Preparation of 4- (bromomethyl) -7-chloro-5-fluoro-1,3-benzothiazole
(7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methanol 166j (130mg, 0.60mmol) was dissolved in dichloromethane (10 mL) and phosphorus tribromide (161mg, 0.60mmol) was added dropwise to the solution at 0 ℃. The mixture was stirred at 0 ℃ for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (10 mL), extracted with dichloromethane (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration gave crude 4- (bromomethyl) -7-chloro-5-fluoro-1,3-benzothiazole 166k (110 mg, yellow solid), yield: 60 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：280.0[M+1] + 。
The eleventh step
Preparation of 7-chloro-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole
4- (bromomethyl) -7-chloro-5-fluoro-1,3-benzothiazole 166k (110mg, 0.39mmol), 4-fluoro-2-methoxy-5-nitroaniline (88mg, 0.47mmol), potassium carbonate (108mg, 0.78mmol) and potassium iodide (32mg, 0.19mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50 ℃ for 2 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (ethyl acetate/petroleum ether = 23%) to give 7-chloro-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 166l (65 mg, white solid) in yield: 42 percent.
MS m/z(ESI)：387.0[M+1] + 。
The twelfth step
Preparation of 5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline
Dissolve 7-chloro-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1,3-benzothiazole 166l (60mg, 0.15mmol), iron powder (43mg, 0.77mmol) and ammonium chloride (41mg, 0.77mmol) in ethanol: water =1:1 (10 mL), and the reaction mixture was stirred at 60 ℃ for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was quenched with water (10 mL), extracted with ethyl acetate (3 × 10 mL), and the combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline 166m (50 mg, brown solid) in yield: 88 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：357.0[M+1] + 。
Thirteenth step
Preparation of dimethyl 4- [ ({ [5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate
5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline 166m (45mg, 0.13mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester (50mg, 0.15mmol) and triethylamine (25mg, 0.25mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, filtration and concentration gave 4- [ ({ [5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 166n (50 mg, yellow solid), yield: 53 percent. The crude product was used directly in the next reaction.
MS m/z(ESI)：598.0[M+1] + 。
Fourteenth step
Preparation of 3- {5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolving dimethyl 4- [ ({ [5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylate 166n (45mg, 0.075mmol) in tetrahydrofuran/water =1:1 (5 mL), then lithium hydroxide (10mg, 0.37mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction was adjusted to pH <7 with 1M HCl, washed with water (10 mL), extracted with ethyl acetate (3X 10 mL), the combined organic layers were washed with brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA) to give 3- {5- [ (7-chloro-5-fluoro-1,3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-079 (17.39 mg, white solid) yield: 41 percent.
MS m/z(ESI)：552.0[M+1] + 。
HPLC:100.00％(214nm),99.50％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.52(s,1H),12.00(s,1H),9.58(s,1H),7.83(d,J＝9.6Hz,1H),7.38(s,1H),7.35(d,J＝7.2Hz,1H),7.13(d,J＝11.6Hz,1H),5.43(s,2H),3.78(s,3H).
Example 80
Preparation of 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-080)
Figure BDA0003608368340001631
First step of
Preparation of 2,3-dicyano diethyl fumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL, 0.07mol) and ethyl 2-cyanoacetate 004a (5 mL, 0.04mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction mixture was stirred under reflux at 90 ℃ for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid precipitated. The reaction was filtered and washed with glacial ethanol to give 2,3-diethyl dicyano fumarate 004b (2.33 g, white solid) in yield: 26 percent.
MS m/z(ESI)：223.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ4.49(q,J＝7.2Hz,4H),1.44(t,J＝7.2Hz,6H).
Second step of
Preparation of 5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester
2,3-dicyano diethyl fumarate 004b (500mg, 2.25mmol) was dissolved in ethanol (10 mL), acethydrazide (218mg, 2.48mmol) and sodium acetate (18mg, 0.23mmol) were added, and the reaction mixture was reacted at 90 ℃ for 0.5 hour. After completion of the reaction, it was cooled to room temperature and extracted with dichloromethane (3X 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 50% ethyl acetate/50% petroleum ether) to give 5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester 004c (333 mg, white solid) with yield: 31 percent.
MS m/z(ESI)：228.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ4.42(q,J＝7.2Hz,2H),4.30(q,J＝7.2Hz,2H),1.40(t,J＝7.2Hz,3H),1.35(t,J＝7.2Hz,3H).
The third step
Preparation of ethyl 3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylate
5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester 004c (390mg, 1.24mmol) was dissolved in dichloromethane (4 mL), 1,1' -carbonyldiimidazole (241mg, 1.49mmol), triethylamine (376 mg, 3.72mmol) and 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyaniline (282mg, 1.24mmol) were added, and the reaction solution was reacted at 25 ℃ for 1.5 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3X 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 5% methanol/95% dichloromethane) to give 3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylic acid ethyl ester 004d (554 mg, white solid) with a yield of 79%.
MS m/z(ESI)：567.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.81(s,1H),7.75(d,J＝7.6Hz,1H),7.15-7.08(m,1H),7.04(brs,2H),6.73(d,J＝11.8Hz,1H),6.61-6.58(m,1H),5.17(d,J＝1.4Hz,2H),4.43(q,J＝7.1Hz,2H),4.29(q,J＝7.1Hz,2H),3.83(s,3H),3.82(s,3H),1.42(t,J＝7.1Hz,3H),1.33(t,J＝7.1Hz,3H).
The fourth step
Preparation of 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylic acid ethyl ester 004d (100mg, 0.18mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (38mg, 0.90mmol) was added, the reaction was stirred at 25 ℃ for 4 hours, after completion of the reaction, the reaction system was made acidic with 2N diluted hydrochloric acid, dichloromethane was added to extract (2X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by preparative column (mobile phase: 0.1% acetic acid/acetonitrile/water) to give 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-3536 zzft 3536-tetrahydro-1H-pyrazolo [3,4 ] pyrimidine-3 d-3926 d ] pyrimidine-3 d-08017 mg, white solid yield.
MS m/z(ESI)：493.1[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,CDCl 3 )δ14.07(s,1H),10.33(s,1H),7.12(dd,J＝18.7,9.3Hz,1H),6.92(d,J＝7.2Hz,1H),6.83(d,J＝11.1Hz,1H),6.59(d,J＝9.1Hz,1H),5.14(s,2H),3.89(s,3H),3.79(s,3H).
Example 81
Preparation of 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4,6-dioxy-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-081)
Figure BDA0003608368340001641
First step of
Preparation of ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4,6-dioxy-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate
Ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4,6-dioxy-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate 073a (100mg, 0.19mmol) was dissolved in acetonitrile (10 mL) followed by the addition of iodomethane (27mg, 0.19mmol) and potassium carbonate (80mg, 0.58mmol). Stir at room temperature for 1h. After the reaction was completed, the pH was adjusted to 2 to 3 by adding 6N hydrochloric acid solution, and then purified by reverse column (acetonitrile/water: 75/25) to obtain the objective product ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4,6-dioxy-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate 073b (15 mg, white solid), yield: 10 percent.
MS m/z(ESI)：535.1[M+1] + 。
The third step
Preparation of 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylic acid
Ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4,6-dioxy-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate 073b (15mg, 0.03mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL), followed by addition of lithium hydroxide monohydrate (4mg, 0.09mmol). The reaction was stirred at room temperature for 1h. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and then the mixture was purified by reverse column (acetonitrile/water) followed by preparative (acetonitrile/0.1% formic acid aqueous solution) to give 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4,6-dioxy-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid Cpd-081 (5 mg, white solid), yield: 27 percent.
MS m/z(ESI)：507.1[M+1] + 。
HPLC:98.37％(214nm),98.57％(254nm).
1 H NMR(400MHz,CD 3 OD)δ7.22(dd,J＝19.2,9.2Hz,1H),6.97(dd,J＝22.4,9.2Hz,2H),6.77(d,J＝9.2Hz,1H),5.09(s,2H),3.90(s,3H),3.85(s,3H),3.79(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-128.78,-142.14,-150.58.
Example 82
Preparation of methyl 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylate (Cpd-082)
Figure BDA0003608368340001651
First step of
Preparation of 2,3-dicyano diethyl fumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL, 0.07mol) and ethyl 2-cyanoacetate 075a (5 mL, 0.04mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction mixture was stirred under reflux at 90 ℃ for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid precipitated. The reaction was filtered and washed with glacial ethanol to give 2,3-diethyl dicyanocarbamate 075b (2.33 g, white solid) in yield: 26 percent.
MS m/z(ESI)：223.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ4.49(q,J＝7.2Hz,4H),1.44(t,J＝7.2Hz,6H).
Second step of
Preparation of 5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester
2,3-dicyano diethyl fumarate 075b (500mg, 2.25mmol) was dissolved in ethanol (10 mL), acethydrazide (218mg, 2.48mmol) and sodium acetate (18mg, 0.23mmol) were added, and the reaction mixture was reacted at 90 ℃ for 0.5 hour. After completion of the reaction, it was cooled to room temperature and extracted with dichloromethane (3X 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 50% ethyl acetate/50% petroleum ether) to give 5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester 075c (333 mg, white solid) in yield: 31 percent.
MS m/z(ESI)：228.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ4.42(q,J＝7.2Hz,2H),4.30(q,J＝7.2Hz,2H),1.40(t,J＝7.2Hz,3H),1.35(t,J＝7.2Hz,3H).
The third step
Preparation of ethyl 3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylate
5-amino-1H-pyrazole-3,4-dicarboxylic acid ethyl ester 075c (390mg, 1.24mmol) was dissolved in dichloromethane (4 mL), 1,1' -carbonyldiimidazole (241mg, 1.49mmol), triethylamine (376mg, 3.72mmol) and 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyaniline (282mg, 1.24mmol) were added, and the reaction solution was reacted at 25 ℃ for 1.5 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3X 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 5% methanol/95% dichloromethane) to give 3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylic acid ethyl ester 075d (554 mg, white solid) with a yield of 79%.
MS m/z(ESI)：567.2[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.81(s,1H),7.75(d,J＝7.6Hz,1H),7.15-7.08(m,1H),7.04(brs,2H),6.73(d,J＝11.8Hz,1H),6.61-6.58(m,1H),5.17(d,J＝1.4Hz,2H),4.43(q,J＝7.1Hz,2H),4.29(q,J＝7.1Hz,2H),3.83(s,3H),3.82(s,3H),1.42(t,J＝7.1Hz,3H),1.33(t,J＝7.1Hz,3H).
The fourth step
Preparation of 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
3- (3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4,5-dicarboxylic acid ethyl ester 075d (307mg, 0.54mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (65mg, 2.71mmol) was added, the reaction was stirred at 25 ℃ for 4 hours, after completion of the reaction, the reaction system was adjusted to acidic with 2N diluted hydrochloric acid, dichloromethane was added to extract (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 5- (5- (((3963 zxft 63-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-3 d ] pyrimidine-3 d as a white solid in yield (58 mg, 3926 d).
MS m/z(ESI)：493.1[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ14.07(s,1H),10.33(s,1H),7.12(dd,J＝18.7,9.3Hz,1H),6.92(d,J＝7.2Hz,1H),6.83(d,J＝11.1Hz,1H),6.59(d,J＝9.1Hz,1H),5.14(s,2H),3.89(s,3H),3.79(s,3H).
The fifth step
Preparation of methyl 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylate
After 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid 075e (58mg, 0.12mmol) was dissolved in dichloromethane (2 mL), oxalyl chloride (150mg, 1.18mmol) and a catalytic amount of N, N-dimethylformamide were added, and the reaction mixture was concentrated and dissolved in dichloromethane (2 mL), and the solution was added dropwise to methanol (2 mL) and reacted at 25 ℃ for 0.5 hour, the reaction mixture was concentrated and purified by a preparative column (mobile phase: 0.1% acetic acid/acetonitrile/water) to obtain 5- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -pyrimidine-3-carboxylic acid 075e (58mg, 0.18mmol), and yield of white pyridine-5- (((34578 zxft) was increased by stirring to obtain white pyridine-3-5- (((345732).
MS m/z(ESI)：507.2[M+1] + 。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.52-7.45(m,1H),7.13(d,J＝7.2Hz,1H),7.05(d,J＝11.6Hz,1H),6.93-6.90(m,1H),4.94(s,2H),3.85(s,3H),3.81(s,3H),3.78(s,3H).
Example 83
Preparation of 5- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -4,6-dioxy-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-083)
Figure BDA0003608368340001661
First step of
Preparation of 5- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) -1H-pyrazole-3,4-dicarboxylic acid diethyl ester
5-amino-1H-pyrazole-3,4-dicarboxylic acid diethyl ester (114mg, 0.50mmol) was dissolved in dichloromethane (4 mL), 1,1' -carbonyldiimidazole (98mg, 0.60mmol), triethylamine (153mg, 1.51mmol) and 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyaniline 101a (150mg, 0.50mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1.5 hours. After completion of the reaction, water quenching was added, extraction was performed with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl acetate = 1/1) to give diethyl 5- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) -1H-pyrazole-3,4-dicarboxylate 101b (200 mg, light yellow solid), yield: 49 percent.
MS m/z(ESI)：552.1[M+1] + 。
Second step of
Preparation of 5- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -4,6-dioxy-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid
Ethyl 5- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) -1H-pyrazole-3,4-dicarboxylate 101b (70mg, 0.13mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (30mg, 1.27mmol) was added, and the reaction was stirred at 25 ℃ for 4 hours. After completion of the reaction, the solution was concentrated and purified by preparative column (mobile phase: 0.1% acetic acid/acetonitrile/water) to give 5- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -4,6-dioxy-1h, 7 h-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid preparation Cpd-083 (5 mg, white solid), yield: 8 percent.
MS m/z(ESI)：478.1[M+1] + 。
HPLC:99.30％(214nm),98.61％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.44(s,3H),12.27(s,3H),9.54(s,1H),8.57(d,J＝5.6Hz,1H),7.99(dd,J＝7.2,2.0Hz,1H),7.89(d,J＝5.6Hz,1H),7.80(t,J＝6.4Hz,2H),7.38(d,J＝7.2Hz,1H),7.13(d,J＝11.6Hz,1H),5.59(s,2H),3.82(s,3H).
19 F NMR(376MHz,d 6 -DMSO)δ-128.30.
Example 84
Preparation of 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-084)
Figure BDA0003608368340001671
First step of
Preparation of Ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4,6-dioxo-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate
5- [ ({ 5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -1H-pyrazole-3,4-dicarboxylic acid diethyl ester 118a (200mg, 0.35mmol) was dissolved in tetrahydrofuran, followed by addition of 4A molecular sieve (50 mg), stirring at room temperature for 30 minutes, and then sodium hydride (85mg, 3.53mmol) was added under ice-bath conditions. The reaction was stirred at room temperature for 3 hours. After completion of the LCMS detection reaction, sodium hydride was quenched by addition of saturated ammonium chloride solution under ice bath, then extracted with dichloromethane (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate, and the residue after removal of the organic solvent by distillation under reduced pressure was subjected to silica gel column (petroleum ether/ethyl acetate = 3/7) to give ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4,6-dioxy-1 h,7 h-pyrazoline [3,4-d ] pyrimidine-3-carboxylate 118b (120 mg, white solid), yield: 80 percent.
MS m/z(ESI)：521.1[M+1] + 。
Second step of
Preparation of ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylate
Ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4,6-dioxy-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylate 073a (100mg, 0.19mmol) was dissolved in acetonitrile (10 mL) followed by the addition of iodomethane (27mg, 0.19mmol) and potassium carbonate (80mg, 0.58mmol). Stir at room temperature for 1h. After the reaction was completed, the pH was adjusted to 2-3 by adding 6N hydrochloric acid solution, and then purified by reverse column (acetonitrile/water: 75/25) to obtain the objective product ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylate 118c (50 mg, white solid), yield: 45 percent.
MS m/z(ESI)：549.4[M+1] + 。
The third step
Preparation of 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylic acid
Ethyl 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylate 118c (50mg, 0.09mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL) followed by lithium hydroxide monohydrate (111mg, 0.27mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction, the solvent was removed by concentration under reduced pressure and then purified by reverse column (acetonitrile/water) followed by preparative (acetonitrile/0.1% aqueous formic acid) purification to give 5- {5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1,7-dimethyl-4,6-dioxapyrazole [3,4-d ] pyrimidine-3-carboxylic acid Cpd-084 (30 mg, white solid), yield: 62 percent.
MS m/z(ESI)：521.1[M+1] + 。
HPLC:97.64％(214nm),97.33％(254nm).
1 H NMR(400MHz,CD 3 OD)δ7.22(dd,J＝19.2,9.2Hz,1H),6.98(dd,J＝19.2,9.2Hz,2H),6.78-6.75(m,1H),5.08(s,2H),4.25(s,3H),3.86(s,3H),3.78(s,3H),3.56(s,3H).
19 F NMR(376MHz,CD 3 OD)δ-128.68,-142.18,-150.58.
Example 85
Preparation of 2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1,3-diketoisoindole-4-carboxylic acid (Cpd-085)
Figure BDA0003608368340001681
First step of
Preparation of 2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1,3-diketoisoindole-4-carboxylic acid
Phenyl-1,2,3-tricarboxylic acid (70mg, 0.33mmol) was dissolved in toluene (5 mL) and triethylamine (33.7 mg, 0.33mmol), 4A molecular sieve (100 mg) were added. The reaction mixture was reacted at 110 ℃ for 3 hours. Then 5- [ (2,3-difluoro-6-methoxyphenyl) methyloxy ] -2-fluoro-4-methoxyaniline 023a (83.48mg, 27mmol) was added. The reaction mixture was reacted at 110 ℃ for 9 hours. After the reaction was completed, the solvent was concentrated under reduced pressure, and purified by preparative HPLC to give 2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1,3-diketoisoindole-4-carboxylic acid Cpd-085 (12 mg, brown solid), yield: 9 percent.
MS m/z(ESI)：488.1[M+1] + 。
HPLC:98.36％(214nm),98.52％(254nm).
1 H NMR(400MHz,CDCl 3 )δ13.50(br 1H),8.74(d,J＝7.3Hz,1H),8.22(d,J＝6.8Hz,1H),8.03(d,J＝7.7Hz,1H),7.13(d,J＝9.6Hz,1H),6.96(d,J＝7.0Hz,1H),6.83(d,J＝11.2Hz,1H),6.60(d,J＝7.9Hz,1H),5.16(d,J＝1.4Hz,2H),3.90(s,3H),3.80(s,3H).
Example 86
Preparation of 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid (Cpd-086)
Figure BDA0003608368340001691
First step of
Preparation of 4-fluoro-5-iodo-2-methoxyphenol
Methyl 4-fluoro-5-iodo-2-methoxybenzoate 024a (500mg, 1.6 mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of methanol (2.5 mL), 1M sodium hydroxide solution (2.5 mL) in that order. The reaction mixture was reacted at 25 ℃ for 2 hours. Then extracted with ethyl acetate (3X 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 4-fluoro-5-iodo-2-methoxyphenol 024b (450 mg, yellow oil) in yield: 93 percent.
1 H NMR(400MHz,CDCl 3 )δ7.23(d,J＝6.2Hz,1H),6.64(d,J＝8.8Hz,1H),3.87(s,3H).
Second step of
Preparation of 1,2-difluoro-3- ((4-fluoro-5-iodo-2-methoxyphenoxy) methyl) -4-methoxybenzene
4-fluoro-5-iodo-2-methoxyphenol 024b (400mg, 1.49mmol) was dissolved in N, N-dimethylformamide (20 mL), and then 2- (chloromethyl) -3,4-difluoro-1-methoxybenzene (287mg, 1.49mmol), potassium carbonate (412mg, 2.98mmol), potassium iodide (124mg, 0.74mmol) were added in that order. The reaction mixture was reacted at 80 ℃ for 2 hours. Then extracted with ethyl acetate (2X 50 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (petroleum ether/ethyl acetate = 70/30) to give 1,2-difluoro-3- ((4-fluoro-5-iodo-2-methoxyphenoxy) methyl) -4-methoxybenzene 024c (500 mg, white solid) in yield: and 69 percent.
MS m/z(ESI)：447.0(M+23)。
The third step
Preparation of 2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,4,5,5-tetramethyl-1,3,2-dioxaborolan
1,2-difluoro-3- ((4-fluoro-5-iodo-2-methoxyphenoxy) methyl) -4-methoxybenzene 024c (500mg, 1.20mmol) was dissolved in dioxane (5 mL). Then bis-pinacolato diboron (450mg, 1.75mmol), potassium acetate (345mg, 3.55mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (85mg, 0.10mmol) were added in this order. The reaction mixture was reacted at 100 ℃ for 16 hours under nitrogen protection. The reaction solution was filtered with suction, and the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give 2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,4,5,5-tetramethyl-1,3,2-dioxaborolan 024d (200 mg, white solid) in yield: 36 percent.
MS m/z(ESI)：424.2[M+1] + 。
The fourth step
Preparation of (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxybenzyl) boronic acid
2- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,4,5,5-tetramethyl-1,3,2-dioxaborolan 024d (150mg, 0.35mmol) was dissolved in acetone (5 mL) and then water (5 mL), sodium periodate (227 mg, 1.06mmol), ammonium acetate (109mg, 1.41mmol) were added in that order. The reaction mixture was reacted at 25 ℃ for 16 hours. Then extracted with ethyl acetate (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate and concentrated to give (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxybenzene) boronic acid 024e (120 mg, yellow solid) in yield: 89 percent.
MS m/z(ESI)：343.1[M+1] + 。
The fifth step
Preparation of methyl 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylate
024e (50mg, 0.15mmol) of (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxybenzene) boronic acid was dissolved in toluene (5 mL), followed by the addition of 1H-indole-3-carboxylic acid methyl ester (52.3mg, 0.3mmol), pyridine (94.9mg, 1.2mmol), 1M lithium bistrimethylsilylamide (0.3 mL) in that order. The reaction mixture was reacted at 100 ℃ for 16 hours. The reaction solution was filtered with suction, and the filtrate was concentrated to give 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid methyl ester 024f (30 mg, yellow oil) by preparative HPLC, yield: 30 percent.
MS m/z(ESI)：473.3[M+1] + 。
The sixth step
Preparation of 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid
Methyl 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylate 024f (30mg, 0.06mmol) was dissolved in tetrahydrofuran (2 mL). Then methanol (2 mL), water (1 mL), and lithium hydroxide (2.87mg, 0.12mmol) were added in this order. The reaction mixture was reacted at 25 ℃ for 1 hour. After the reaction was complete, the pH was adjusted to 2 to 3 with 1M hydrochloric acid solution. The reaction was concentrated and purified by preparative HPLC to give 1- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid Cpd-086 (11 mg, white solid) in yield: 41 percent.
MS m/z(ESI)：459.1[M+1] + 。
HPLC:100.00％(214nm),98.94％(254nm).
1 H NMR(400MHz,MeOD)δ8.26(d,J＝8.4Hz,1H),7.52-7.49(m,1H),7.41-7.38(m,1H),7.32-7.28(m,2H),7.24-7.20(m,1H),7.12-7.09(m,1H),6.80(s,1H),5.19(s,2H),3.92(s,3H),3.77(s,3H).
Examples 87 and 88
Preparation of 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid (Cpd-087) and 2- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid (Cpd-088)
Figure BDA0003608368340001701
First step of
Preparation of 2,3-difluoro-6-methoxybenzaldehyde
A2M lithium diisopropylamide solution (3.82mL, 7.64mmol) was dissolved in tetrahydrofuran (10 mL) under a nitrogen atmosphere, the temperature was lowered to-78 ℃, a tetrahydrofuran (3 mL) solution of 1,2-difluoro-4-methoxybenzene 025a (1000mg, 6.94mmol) was added dropwise, the reaction solution was stirred at-78 ℃ for 1 hour, then N, N-dimethylformamide (558mg, 7.64mmol) was added dropwise at-78 ℃, and the reaction solution was stirred at-78 ℃ for 1 hour. To the reaction solution were added 15mL of acetic acid and 50mL of water, extracted with ethyl acetate (3 × 50 mL), the organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by a silica gel column (dichloromethane/methanol = 30/1) to give the title product 2,3-difluoro-6-methoxybenzaldehyde 025b (1050 mg, yellow solid) in yield: 79 percent.
MS m/z(ESI)：173.1[M+1] + .
1 H NMR(400MHz,CDCl 3 )δ10.44-10.37(m,1H),7.34(dd,J＝18.0,9.2Hz,1H),6.72-6.68(m,1H),3.92(s,3H).
Second step of
Preparation of 2,3-difluoro-6-methoxybenzyl alcohol
2,3-difluoro-6-methoxybenzaldehyde 025b (1000mg, 5.81mmol) was dissolved in toluene (10 mL), and under an ice-water bath 1.1mL0.1N sodium hydroxide solution was added followed by sodium borohydride (264 mg, 6.98mmol). The reaction solution was stirred at 25 ℃ for 4 hours. To the reaction was added 50mL of water, extracted with ethyl acetate (3X 30 mL), the organic phases combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate concentrated to give the title product 2,3-difluoro-6-methoxybenzyl alcohol 025c (980 mg, yellow oil), yield: 87 percent.
MS m/z(ESI)：157.1[M+1] + .
The third step
Preparation of 2,3-difluoro-6-methoxybenzyl chloride
2,3-difluoro-6-methoxybenzyl alcohol 025c (980mg, 5.63mmol) was dissolved in toluene (8 mL), and concentrated hydrochloric acid (4.7 mL) was added. The reaction mixture was stirred at 50 ℃ for 6 hours. To the reaction was added 50mL of water, extracted with ethyl acetate (3 × 30 mL), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated to give the title product 2,3-difluoro-6-methoxybenzyl chloride 025d (900 mg, yellow oil), yield: 75 percent.
The fourth step
Preparation of 2- (5-bromo-2-methoxyphenoxymethyl) -3,4-difluoro-1-methoxybenzene
5-bromo-2-methoxyphenol (300mg, 1.48mmol) was dissolved in N, N-dimethylformamide (5 mL), and 2,3-difluoro-6-methoxybenzyl chloride 025d (428mg, 2.22mmol), potassium carbonate (614mg, 4.44mmol) and potassium iodide (369mg, 2.22mmol) were added. The reaction solution was stirred at 80 ℃ for 6 hours. To the reaction solution was added 100mL of water, extracted with ethyl acetate (3 × 50 mL), the organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by a silica gel column (petroleum ether/ethyl acetate = 2/1) to give the title product 2- (5-bromo-2-methoxyphenoxymethyl) -3,4-difluoro-1-methoxybenzene 025e (480 mg, pale yellow solid), yield: 81 percent.
MS m/z(ESI)：381.0[M+1] + .
1 H NMR(400MHz,DMSO)δ7.53-7.45(m,1H),7.30(d,J＝2.4Hz,1H),7.10(dd,J＝8.8,2.4Hz,1H),6.95-6.88(m,2H),5.06(d,J＝1.2Hz,2H),3.83(s,
3H),3.70(s,3H).
The fifth step
Preparation of methyl 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylate
Methyl 1H-indazole-6-carboxylate (108mg, 0.62mmol) was dissolved in N, N-dimethylacetamide (3 mL) under a nitrogen atmosphere, 2- (5-bromo-2-methoxyphenoxymethyl) -3,4-difluoro-1-methoxybenzene 025e (200mg, 0.56mmol), copper (I) iodide (22mg, 0.11mmol), trans-cyclohexane-1,2-diamine (26mg, 0.22mmol), and potassium phosphate (357mg, 1.68mmol) were added, and the reaction was stirred in a microwave reactor at 130 ℃ for 2 hours. To the reaction solution was added 50mL of water, extracted with ethyl acetate (3 × 30 mL), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by a silica gel column (petroleum ether/ethyl acetate = 1/1) to give the title product, methyl 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylate 025f (180 mg, light yellow solid), yield: 70 percent.
MS m/z(ESI)：455.1[M+1] + .
The sixth step
Preparation of 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid (Cpd-087) and 2- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid (Cpd-088)
Methyl 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylate 025f (100mg, 0.22mmol) was dissolved in a solution of methanol/water =4/1 (3 mL), lithium hydroxide monohydrate (47mg, 1.1mmol) was added, and the reaction solution was stirred at 25 ℃ for 2 hours. The pH of the reaction solution was adjusted to pH =6 with 1N hydrochloric acid, extracted with ethyl acetate (3 × 30 mL), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by preparative HPLC (acetonitrile/water (0.1% formic acid)) to give a crude product (60 mg), then purified with SFC (carbon dioxide/methanol (0.1% diethylamine)) to give the title product 1- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid Cpd-087 (33 mg, white solid), yield: 34 percent; 2- {3- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid Cpd-088 (16.3 mg, light yellow solid), yield: 16 percent.
Cpd-087 data:
MS m/z(ESI)：441.1[M+1] + .
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ8.38(d,J＝0.8Hz,1H),8.23(d,J＝0.8Hz,1H),7.87(d,J＝8.4Hz,1H),7.80(dd,J＝8.4,1.2Hz,1H),7.52-7.43(m,2H),7.30(dd,J＝8.8,2.4Hz,1H),7.20(d,J＝8.8Hz,1H),6.93-6.89(m,1H),5.14(s,2H),3.82(s,3H),3.80(s,3H).
cpd-088 data:
MS m/z(ESI)：441.1[M+1] + .
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ9.10(s,1H),8.28(s,1H),7.88(d,J＝2.4Hz,1H),7.75(d,J＝8.8Hz,1H),7.66(dd,J＝8.4,3.2Hz,2H),7.50(dd,J＝19.6,9.6Hz,1H),7.16(d,J＝8.8Hz,1H),6.98-6.92(m,1H),5.22(s,2H),3.86(s,3H),3.80(s,3H).
example 89
Preparation of 3- (2-fluoro-5- (((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-089)
Figure BDA0003608368340001721
First step of
Preparation of N- (3-fluoro-2-methylphenyl) acetamide
3-fluoro-2-methylaniline 133a (5.00g, 39.95mmol) was dissolved in dichloromethane (30 mL), triethylamine (8.07g, 79.81mmol) and acetyl chloride (3450mg, 43.95mmol) were added at 0 ℃ and stirred at 0 ℃ for 1 hour. After the reaction was complete, quench with water (20 mL) and extract with DCM (3X 50 mL), wash the organic phase with brine, dry over anhydrous sodium sulfate and filter. The filtrate was concentrated and dried to give N- (3-fluoro-2-methylphenyl) acetamide 133b (6.00 g, white solid) in yield: 81 percent. MS m/z (ESI): 168.2[ 2 ] M +1] + 。
Second step of
Preparation of N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide
N- (3-fluoro-2-methylphenyl) acetamide 133b (4.00g, 23.93mmol) was dissolved in concentrated sulfuric acid (24 mL), fuming nitric acid (2.78g, 28.71mmol) was added dropwise at 0 deg.C, and then stirred at 0 deg.C for 1 hour. After the reaction was completed, 50mL of water was added and extracted with dichloromethane (3 × 50 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to obtain N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide 133c (3.00 g, yellow solid) in yield: 56 percent.
MS m/z(ESI)：213.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ10.04(s,1H),7.86(dd,J＝9.2,5.6Hz,1H),7.32(t,J＝8.8Hz,1H),2.17(d,J＝2.4Hz,3H),2.06(s,3H).
The third step
Preparation of 3-fluoro-2-methyl-6-nitroaniline
Concentrated sulfuric acid (20 mL) was added dropwise to a solution of N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide 133c (3.00g, 14.08mmol) in methanol (20 mL) and stirred at 90 ℃ for 4 hours. After completion of the reaction, the reaction system was concentrated, adjusted to pH 8-9 with 3M sodium hydroxide, and then extracted with methylene chloride (3X 20 mL)The organic phase is dried by anhydrous sodium sulfate and then filtered, and the filtrate is concentrated and dried to obtain 3-fluoro-2-methyl-6-nitroaniline 133d (2.80 g, yellow solid), the yield: 90 percent. MS m/z (ESI): 171.1[ 2 ] M +1] + 。
The fourth step
Preparation of 4-fluoro-3-methylbenzene-1,2-diamine
To a solution of 3-fluoro-2-methyl-6-nitroaniline 133d (1.29g, 7.60mmol) in ethanol (10 mL) were added a saturated aqueous ammonium chloride solution (10 mL) and iron powder (1.27g, 22.80mmol), and the mixture was stirred at 80 ℃ for 1 hour. After the reaction was completed, the reaction mixture was filtered while it was hot, the filtrate was extracted with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to obtain 4-fluoro-3-methylbenzene-1,2-diamine 133e (517 mg, brown solid), yield: 49 percent. MS m/z (ESI): 141.2[ M ] +1] + 。
1 H NMR(400MHz,CDCl 3 )δ6.52(dd,J＝8.4,5.4Hz,1H),6.39(t,J＝8.8Hz,1H),3.35(s,4H),2.09(d,J＝1.6Hz,3H).
The fifth step
Preparation of 6-fluoro-5-methylquinoxaline
To a solution of 4-fluoro-3-methylbenzene-1,2-diamine 133e (517mg, 3.69mmol) in ethanol (10 mL) was added glyoxal (1.07g, 7.38mmol,40% aqueous solution) and the reaction was stirred at room temperature for 3 hours. After the reaction was completed, extraction was performed with ethyl acetate (3 × 20 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain 6-fluoro-5-methylquinoxaline 133f (160 mg, white solid) with a yield: 27 percent.
MS m/z(ESI)：163.0[M+1] + 。
1 H NMR(400MHz,CDCl 3 )δ8.86(d,J＝1.8Hz,1H),8.81(d,J＝1.8Hz,1H),7.96(dd,J＝9.2,5.8Hz,1H),7.54(t,J＝9.2Hz,1H),2.70(d,J＝2.4Hz,3H).
The sixth step
Preparation of 5- (bromomethyl) -6-fluoroquinoxaline
To a solution of 6-fluoro-5-methylquinoxaline 133f (160mg, 0.99mmol) in carbon tetrachloride (5 mL) were added 2,2' -azobis (2-methylpropionitrile) (24mg, 0.10mmol) and N-bromosuccinimide (176mg, 0.99mmol), and stirred at 80 ℃ for 3 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3X 10 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated and dried to obtain 133g (208 mg, yellow solid) of 5- (bromomethyl) -6-fluoroquinoxaline, yield: 87 percent.
MS m/z(ESI)：241.0[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.11(d,J＝1.8Hz,1H),9.04(d,J＝1.8Hz,1H),8.23(dd,J＝9.4,5.8Hz,1H),7.89(t,J＝9.4Hz,1H),5.21(d,J＝1.4Hz,
2H).
Seventh step
Preparation of 6-fluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline
To 4-fluoro-2-methoxy-5-nitrophenol (178mg, 0.95mmol) in acetonitrile (10 mL) were added potassium carbonate (179mg, 1.29mmol) and 133g of 5- (bromomethyl) -6-fluoroquinoxaline (208mg, 0.86mmol), and the reaction mixture was stirred at 70 ℃ for 3 hours. After the reaction, water quenching was added, extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, the filtrate was concentrated and then purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to obtain 6-fluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 133h (227 mg, white solid), yield: 76 percent.
MS m/z(ESI)：3488.1[M+1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.05(dd,J＝14.0,1.8Hz,2H),8.29(dd,J＝9.4,6.0Hz,1H),8.02(d,J＝7.4Hz,1H),7.93(t,J＝9.4Hz,1H),7.25(d,J＝13.4Hz,1H),5.75(s,2H),3.84(s,3H).
Eighth step
Preparation of 2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyaniline
To a solution of-fluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 133h (177mg, 0.51mmol) in ethanol (3 mL) and water (3 mL) were added iron powder (85mg, 1.53mmol) and ammonium chloride (82mg, 1.53mmol) and the reaction mixture was taken up in 8Stirred at 0 ℃ for 2 hours. After the reaction, the reaction mixture was filtered while hot, the filtrate was extracted with dichloromethane (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated and dried to obtain 2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyaniline 133i (155 mg, yellow solid) in yield: 96 percent. MS m/z (ESI): 318.2[ 2 ] M +1] + 。
1 H NMR(400MHz,DMSO-d 6 )δ9.05-9.01(m,2H),8.26(dd,J＝9.3,5.9Hz,1H),7.90(t,J＝9.3Hz,1H),6.76(d,J＝12.5Hz,1H),6.67(d,J＝8.9Hz,1H),5.53(s,2H),4.66(s,2H),3.56(s,3H).
The ninth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate Triethylamine (60mg, 0.59mmol) and methyl 4- ((phenoxycarbonyl) amino) thiophene dimethyl-2,3-dicarboxylate (145mg, 0.43mmol) were added to a solution of 2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyaniline 133i (125mg, 0.39mmol) in tetrahydrofuran (5 mL) and stirred at 25 ℃ for 5 hours. After the reaction, water quenching was added, extraction was performed with dichloromethane (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate = 1/1) to give 4- (3- (2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 133j (148 mg, white solid) with yield: 67%.
MS m/z(ESI)：559.1[M+1] + 。
The tenth step
Preparation of 3- (2-fluoro-5- (((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of 4- (3- (2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 133j (118mg, 0.21mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (15mg, 0.63mmol) and reacted at 25 ℃ for 2 hours. After the reaction was completed, 2N diluted hydrochloric acid was added dropwise to adjust pH =6-7, extraction was performed with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by preparative column to give 3- (2-fluoro-5- (((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-089 (34 mg, white solid) with a yield of 31%.
MS m/z(ESI)：513.0[M+1] + 。
HPLC:99.02％(214nm),96.55％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ9.03(dd,J＝11.5,1.8Hz,1H),8.29(dd,J＝9.3,5.9Hz,1H),7.92(t,J＝9.2Hz,1H),7.38(d,J＝7.3Hz,1H),7.23(s,1H),7.11(d,J＝11.5Hz,1H),5.55(s,2H),3.75(s,3H).
Example 90
Preparation of 3- (5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-090)
Figure BDA0003608368340001751
First step of
Preparation of 1,2-difluoro-3-methyl-4-nitrobenzene
Compound 135a (10g, 78.1mmol) was dissolved in concentrated sulfuric acid (50 mL), and concentrated nitric acid (4.92g, 78.1mmol) was slowly added dropwise at-10 ℃ to react at 0 ℃ for 1 hour. The reaction was poured into ice water and saturated NaHCO was used 3 The solution was adjusted to pH 7, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (PE) to give 1,2-difluoro-3-methyl-4-nitrobenzene 135b (10 g, yellow oily liquid) in yield: 74 percent.
1H NMR(400MHz,CDCl3)δ7.86–7.82(m,1H),7.20–7.14(m,1H),2.56(d,J＝2.6Hz,3H).
Second step of
Preparation of 3,4-difluoro-2-methylaniline
Compound 135b (10g, 57.8mmol) was dissolved in MeOH/NH 4 Cl =5:1 solution (100 mL), reduced iron powder (16.14g, 289mmol) was added and reacted at room temperature for 2h. Filtering, concentrating the filtrate under reduced pressure, dissolving the residue in water, and adding ethyl acetate (3)X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 3,4-difluoro-2-methylaniline 135c (6 g, yellow oily liquid), yield: 72.5 percent.
MS m/z(ESI)：144.2[M+1] + .
The third step
Preparation of N- (3,4-difluoro-2-methylphenyl) acetamide
Compound 135c (6 g, 42mmol) and triethylamine (12.7g, 126mmol) were dissolved in dry DCM (60 mL) and acetyl chloride (3.9g, 50.4mmol) was added dropwise under ice bath conditions and reacted at room temperature for 1h. The reaction was quenched with water (50 mL), extracted with dichloromethane (3X 50 mL), washed with saturated NaCl solution and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 70%) to give N- (3,4-difluoro-2-methylphenyl) acetamide 135d (3.3 g, yellow solid) in yield: 42.9 percent.
MS m/z(ESI)：186.2[M+1] + .
The fourth step
Preparation of N- (3,4-difluoro-2-methyl-6-nitrophenyl) acetamide
Compound 135d (1.5g, 8.1mmol) was dissolved in concentrated sulfuric acid (30 mL), and concentrated nitric acid (0.7 mL) was slowly added dropwise at-10 ℃ to react at 0 ℃ for 1 hour. The reaction was poured into ice water and saturated NaHCO was used 3 The solution was adjusted to pH 7, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 50%) to give N- (3,4-difluoro-2-methyl-6-nitrophenyl) acetamide 135e (1 g, white solid) in yield: 53.8 percent.
MS m/z(ESI)：231.0[M+1] + .
The fifth step
Preparation of 3,4-difluoro-2-methyl-6-nitroaniline
Compound 135e (1g, 4.3 mmol) was dissolved in absolute ethanol (20 mL), followed by dropwise addition of concentrated sulfuric acid (3 mL) and reaction at 90 ℃ for 4 hours. Concentrated under reduced pressure, the residue obtained is dissolved in water and saturated NaHCO is used 3 The solution was adjusted to pH 7, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure to obtain 3,4-difluoro-2-methylYl-6-nitroaniline 135f (800 mg, yellow solid), yield: 97.9 percent.
MS m/z(ESI)：189.1[M+1] + .
The sixth step
Preparation of 4,5-difluoro-3-methylbenzene-1,2-diamine
Compound 135f (2.3g, 12.2mmol) is dissolved in MeOH/NH 4 Cl =5:1 solution (60 mL), reduced iron powder (6.81g, 122mmol) was added. The reaction was carried out at room temperature for 16h. After filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was dissolved in water, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl, and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 60%) to give 4,5-difluoro-3-methylbenzene-1,2-diamine 135g (1.17 g, yellow solid), yield: 61 percent.
MS m/z(ESI)：159.2[M+1] + .
Seventh step
Preparation of 6,7-difluoro-5-methylquinoxaline
135g (500mg, 3.16mmol) of the compound is dissolved in HF (10 mL), and an aqueous glyoxal solution (551mg, 9.48mmol) is added dropwise at 0 ℃ to react for 1h at 0 ℃. The reaction was poured into water and saturated NaHCO was used 3 The solution was adjusted to pH 7, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 20%) to give 6,7-difluoro-5-methylquinoxaline 135h (150 mg, white solid) in yield: 26.4 percent.
MS m/z(ESI)：181.2[M+1] + .
Eighth step
Preparation of 5- (bromomethyl) -6,7-difluoroquinoxaline
Compound 135h (150mg, 0.83mmol) was dissolved in carbon tetrachloride (50 mL) and AIBN (13.4 mg, 0.08mmol) and NBS (148mg, 0.83mmol) were added and reacted at 80 ℃ for 2h. The reaction solution was poured into water, extracted with dichloromethane, washed with saturated NaCl solution, and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 20%) to give 5- (bromomethyl) -6,7-difluoroquinoxaline 135i (155 mg, white solid) in yield: 71.9 percent.
MS m/z(ESI)：259.1[M+1] + .
The ninth step
Preparation of 6,7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoxaline
Compound 135i (70mg, 0.27mmol) and 4-fluoro-2-methoxy-5-nitrophenol (76mg, 0.41mmol) were dissolved in ACN (10 mL), potassium carbonate (112mg, 0.81mmol) was added, and reacted at 80 ℃ for 2h. The reaction solution was poured into water, extracted with ethyl acetate, washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 35%) to give 6,7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoxaline 135j (82 mg, white solid), yield: 78.7 percent.
MS m/z(ESI)：366.0[M+1] + .
The tenth step
Preparation of 5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyaniline
Compound 135j (80mg, 0.22mmol) was dissolved in MeOH/NH4Cl =5:1 solution (6 mL), and reduced iron powder (122mg, 2.2mmol) was added and reacted at room temperature for 1h. Filtration and concentration of the filtrate under reduced pressure, the resulting residue was dissolved in water, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 135k (65 mg, light brown solid) in yield: 80 percent.
MS m/z(ESI)：336.1[M+1] + .
The eleventh step
Preparation of 4- (3- (5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
Compound 135k (60mg, 0.18mmol) and triethylamine (55mg, 0.54mmol) were dissolved in dry THF (5 mL), and methyl 4- ((phenoxycarbonyl) amino) thiophenedimethyl-2,3-dicarboxylate (90mg, 0.27mmol) was added and reacted at room temperature for 16h. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: PE = 60%) to give dimethyl 4- (3- (5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 135l (75 mg, light yellow solid), yield: and 69 percent.
MS m/z(ESI)：577.2[M+1] + .
The twelfth step
Preparation of 3- (5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
135l of compound (75mg, 0.13mmol) in THF/H 2 To a mixed solution of O =3:1 (8 mL), lithium hydroxide (1695 mg, 0.65mmol) was added and reacted at room temperature for 2h. The reaction mixture was concentrated under reduced pressure, and the residue obtained was subjected to prep-HPLC (ACN: H) 2 O (0.2% FA) = 55%) was purified to give 3- (5- ((6,7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d]Pyrimidine-5-carboxylic acid Cpd-090 (8.5 mg, white solid), yield: 12.3 percent.
MS m/z(ESI)：531.1[M+1] + .
1 H NMR(400MHz,DMSO-d6)δ14.53(s,1H),11.99(s,1H),9.04(d,J＝1.8Hz,1H),9.03(d,J＝1.8Hz,1H),8.33–8.26(m,1H),7.40–7.31(m,2H),7.16–7.12(m,1H),5.59(s,2H),3.78(s,3H).
19 F NMR(376MHz,DMSO-d6)δ-127.57(d,J＝7.6Hz,1H),-130.36(dd,J＝23.0,3.8Hz,1H),-132.22(dd,J＝23.0,3.6Hz,1H).
Example 91
5-acetyl-3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340001771
First step of
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methoxy-N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid 177a (1g, 2mmol) was dissolved in N, N-dimethylformamide (10 mL). N, O-dimethylhydroxylamine hydrochloride (230mg, 2.4 mmol) was then added at 25 ℃; propylphosphoric anhydride (1.91g, 6 mmol); n, N-diisopropylethylamine (260mg, 2mmol). The reaction mixture was reacted at 90 ℃ for 1 hour. Then, the mixture was extracted repeatedly with water (50 mL) and dichloromethane (50 mL) three times, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate: 60/40) to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methoxy-N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide 177b (1 g, yellow solid) in 80% yield.
MS m/z(ESI)：552(M+1)。
Second step of
Preparation of 5-acetyl-3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methoxy-N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide 177b (1000mg, 1.8 mmol) was dissolved in tetrahydrofuran (10 mL). Methyl magnesium bromide (320mg, 2.7 mmol) was then added at 0 ℃. The reaction mixture was reacted at 0 ℃ for 2 hours. After the reaction is finished, the product is directly concentrated, and is subjected to primary purification by a reverse column (water: acetonitrile = 5) to obtain 5-acetyl-3- (5- (((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -diketone Cpd-094 (300 mg, yellow solid) after preparation and purification, wherein the yield is 27.28%.
MS m/z(ESI)：507.0(M+1)。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ11.67(s,1H),7.52–7.44(m,1H),7.28(s,1H),7.22(d,J＝7.4Hz,1H),7.09(d,J＝11.4Hz,1H),6.91(d,J＝9.4Hz,1H),4.95(s,2H),3.80(d,J＝6.0Hz,6H),2.74(s,3H).
Example 92
3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothienyl [3,4-d ] pyrimidine-5-carboxamide
Figure BDA0003608368340001781
Preparation of 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothienyl [3,4-d ] pyrimidine-5-carboxamide
3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-064 (200mg, 0.3865mmol) was dissolved in N, N-dimethylformamide (10 mL), and 1-propylphosphoric acid cyclic anhydride (368.93mg, 1.1595mmol), N, N-diisopropylethylamine (149.85mg, 1.1595mmol), methylamine hydrochloride (31.32mg, 0.4638mmol) were added in this order at room temperature. The reaction was carried out at 90 ℃ for 2 hours under nitrogen. After the reaction was completed, the reaction mixture was concentrated. Then extracted with ethyl acetate and concentrated, and the concentrate was purified by preparative column (water/acetonitrile = 50/50) to give 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothienyl [3,4-d ] pyrimidine-5-carboxamide Cpd-096 (64.48 mg, white solid), yield: 31.44 percent
MS m/z(ESI)：531.1(M+1)。
HPLC:100％(214nm),98.83％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.80(s,1H),10.25(d,J＝5.0Hz,1H),9.50(s,1H),8.30(dd,J＝9.0,5.2Hz,1H),7.57–7.47(m,1H),7.40(d,J＝7.4Hz,1H),7.22(s,1H),7.10(d,J＝11.6Hz,1H),5.44(s,2H),3.77(s,3H),2.86(d,J＝4.8Hz,3H).
Example 93
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide
Figure BDA0003608368340001791
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide
To a solution of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid 177a (200mg, 0.39mmol) in dichloromethane (5 mL) was added oxalyl chloride (150mg, 1.18mmol) and a catalytic amount of N, N-dimethylformamide dropwise, and the reaction was stirred at 25 ℃ for 0.5 hour. After completion of the reaction monitored by TLC, the reaction solution was concentrated. The concentrate was dissolved in methylene chloride (5 mL), triethylamine (60mg, 0.59mmol) and methylamine hydrochloride (32mg, 0.47mmol) were added, and the reaction was stirred at 25 ℃ for 1 hour. After completion of the reaction monitored by LCMS, water was added to quench, extracted with dichloromethane (3 × 20 mL), and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified on preparative column to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -N-methyl-2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxamide Cpd-097 (150 mg, white solid), yield: 73 percent.
MS m/z(ESI)：522.1(M+1)。
HPLC:98.48％(214nm),96.00％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.79(s,1H),10.25(q,J＝4.5Hz,1H),7.49(dd,J＝19.6,9.6Hz,1H),7.26(d,J＝7.4Hz,1H),7.21(s,1H),7.11(d,J＝11.4Hz,1H),6.93-6.90(m,1H),4.95(s,2H),3.81(s,3H),3.80(s,3H),2.86(d,J＝4.4Hz,3H).
Example 94
3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-sodium formate
Figure BDA0003608368340001792
Preparation of sodium 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylate
To a solution of 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-064 (200mg, 0.39mmol) in water (10 mL) was added sodium hydroxide (15mg, 0.39mmol) and the reaction was stirred at 25 ℃ for 0.5 h. The reaction solution was lyophilized to give 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-sodium formate Cpd-098 (200 mg, white solid), yield: 96 percent.
MS m/z(ESI)：518.0。
HPLC:100％(214nm),98.85％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.26(s,1H),9.52(s,1H),8.29(dd,J＝8.8,5.2Hz,1H),7.50(t,J＝9.2Hz,1H),7.28(d,J＝7.2Hz,1H),7.05(d,J＝11.6Hz,1H),6.56(s,1H),5.45(s,2H),3.74(s,3H).
Example 95
3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid choline salt
Figure BDA0003608368340001801
Preparation of choline 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylate salt
To a solution of 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid, cpd-064 (200mg, 0.39mmol) in water (10 mL) was added choline hydroxide (47mg, 0.39mmol), and the reaction was stirred at 25 ℃ for 0.5 h. The reaction was lyophilized to give 3- (2-fluoro-5- ((5-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid choline salt Cpd-099 (232 mg, white solid) in yield: 97 percent.
MS m/z(ESI)：518.0。
HPLC:98.48％(214nm),99.57％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ11.25(s,1H),9.52(s,1H),8.29(dd,J＝9.0,5.2Hz,1H),7.50(t,J＝9.4Hz,1H),7.28(d,J＝7.4Hz,1H),7.06(d,J＝11.6Hz,1H),6.55(s,1H),5.45(s,2H),3.84-3.83(m,2H),3.75(s,3H),3.53–3.47(m,1H),3.41–3.39(m,2H),3.10(s,9H).
Example 95
3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene (3,4-d) pyrimidine-5-carboxylic acid
Figure BDA0003608368340001802
First step of
Preparation of 6-ethoxy-2,3-difluorobenzyl alcohol
6-ethoxy-2,3-difluorobenzaldehyde 181a (420mg, 2.26mmol) was dissolved in methanol (6 mL) and sodium borohydride (428.7mg, 11.3mmol) was added portionwise. The reaction solution was stirred at 80 ℃ for 4 hours. After the reaction was complete, the reaction was quenched by addition of a small amount of water (2 mL), spun dry and the crude product was purified on a silica gel column (methanol/dichloromethane = 8%) to give (6-ethoxy-2,3-difluorophenyl) methanol 181b (280 mg, colorless oil), yield: 62.0 percent.
MS m/z(ESI)：171.1(M+H-18)。
Second step of
Preparation of 2- (bromomethyl) -1-ethoxy-3,4-difluorobenzene
(6-ethoxy-2,3-difluorophenyl) methanol 181b (350mg, 1.86mmol), tetrabromomethane (925.2mg, 2.8mmol) and triphenylphosphine (731.8mg, 2.8mmol) were dissolved in dichloromethane (10 mL). The reaction was stirred at 25 ℃ for 3 hours under hydrogen protection. After the reaction is finished. The reaction was purified by silica gel column (petroleum ether/ethyl acetate = 3%) to give 2- (bromomethyl) -1-ethoxy-3,4-difluorobenzene 181c (300 mg, white solid) in yield: 63.25%.
1 H NMR(400MHz,DMSO-d 6 ))δ7.45–7.35(m,1H),6.87(m,1H),4.61(d,J＝1.4Hz,2H),4.13(m,2H).
The third step
Preparation of N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (bromomethyl) -1-ethoxy-3,4-difluorobenzene 181c (300mg, 1.19mmol), 4-fluoro-2-methoxy-5-nitroaniline (266.9mg, 1.43mmol) and potassium carbonate (329.8mg, 2.39mmol) were dissolved in N, N-dimethylformamide (4 mL) and stirred at 80 ℃ for 2h under nitrogen. After completion of the reaction, water (40 mL) was added to the reaction system to quench it, extracted with EA (3X 5 mL), the organic phase was dried over anhydrous sodium sulfate, and the organic phase was concentrated. The crude product was purified on a silica gel column (petroleum ether: ethyl acetate = 2:1) to give N- ((6-ethoxy-2,3-difluorophenyl) methyl) -4-fluoro-2-methoxy-5-nitroaniline 181d (350 mg, yellow solid), yield: 75.6 percent.
MS m/z(ESI)：357.1(M+H)。
The fourth step
Preparation of N1- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine
N- ((6-ethoxy-2,3-difluorophenyl) methyl 0-4-fluoro-2-methoxy-5-nitroaniline (400mg, 1.12mmol) was dissolved in ethanol water =4:1 (10 mL), ammonium chloride (328.4 mg, 5.61mmol) iron powder (314.4 mg, 5.61mmol) was added and the reaction mixture was stirred at 80 ℃ for 2 hours after the end of the reaction, filtration, the filter cake was washed with ethanol, concentrated and the crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 1:1) to give 1-N- ((6-ethoxy-2,3-difluorophenyl) methyl) -4-fluoro-6-methoxybenzene-1,3-diamine 181e (350 mg, yellow solid) in 82% yield.
MS m/z(ESI)：327.1(M+H)。
The fifth step
Preparation of 4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylic acid dimethyl ester
1-N- ((6-ethoxy-2,3-difluorophenyl) methyl) -4-fluoro-6-methoxybenzene-1,3-diamine (150mg, 0.46mmol), 4- ((phenoxycarbonyl) amino) thiophene-2,3 methyl dicarboxylate (154mg, 0.46mmol) was dissolved in THF (6 mL), and triethylamine (278.6 mg, 2.76mmol) was added dropwise to the system. The reaction was carried out at 25 ℃ for 8 hours. After the reaction was completed, concentration was performed, and the crude product was purified by silica gel chromatography (pure ethyl acetate) to obtain 4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylic acid dimethyl ester 181f (200 mg, yellow solid), yield: 72.1 percent.
MS m/z(ESI)：568.2(M+H)。
The sixth step
Preparation of 3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene (3,4-d) pyrimidine-5-carboxylic acid
4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylic acid dimethyl ester 181f (150mg, 0.26mmol) lithium hydroxide monohydrate (22.18mg 0.53mmol) was dissolved in methanol: tetrahydrofuran (tetrahydrofuran): water =1 (3 mL), the reaction mixture was stirred at 25 ℃ for 2 hours. After the reaction was complete, after concentration, the crude product was purified by silica gel chromatography (dichloromethane: methanol =10: 1) to give 3- (5- ((6-ethoxy-2,3-difluorophenyl) methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1H-thiophene (3,4-d) pyrimidine-5-carboxylic acid Cpd-100 (114 mg, yellow solid), yield: 82.7 percent.
MS m/z(ESI)：521.7(M+H).
HPLC:100.00％(214nm),100.00％(254nm).
1 H NMR(400MHz,DMSO-d 6 ))δ11.99(s,1H),7.31(m,2H),6.98-6.95(m,1H),6.83-6.77(m,2H),4.97(s,1H),4.25(s,2H),4.06(m,2H),3.84(s,3H),1.29(t,J＝6.6Hz,3H).
Example 96
3- (5- ((2,3-difluoro-6-isopropoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001821
First step of
Preparation of 1,2-difluoro-4-isopropoxybenzene
3,4-difluorophenol 182a (5g, 38.4 mmol) was dissolved in acetone (50 mL), 2-iodopropane (7.83g, 46.1mmol) and potassium carbonate (10.61g, 76.8mmol) were added to the solution, the reaction solution was stirred at 56 ℃ overnight, after completion of the reaction, the reaction solution was suction-filtered, and the filtrate was concentrated and purified by silica gel column (petroleum ether) to give 1,2-difluoro-4-isopropoxybenzene 182b (3.9 g, yellow oily liquid) in 58.8% yield.
1 H NMR(400MHz,CDCl 3 )δ7.03(dd,J＝19.2,9.2Hz,1H),6.69(ddd,J＝12.2,6.6,3.0Hz,1H),6.60–6.54(m,1H),4.43(dt,J＝12.1,6.1Hz,1H),1.31(d,J＝6.1Hz,6H).
Second step of
Preparation of 2,3-difluoro-6-isopropoxybenzaldehyde
1,2-difluoro-4-isopropoxybenzene 182b (3.9g, 22.7mmol) was dissolved in tetrahydrofuran (20 mL), a 2M solution of lithium diisopropylamide in tetrahydrofuran (13mL, 24.5mmol) was slowly added dropwise at-78 deg.C, the reaction was stirred at this temperature for half an hour, and then DMF (3.3g, 45.4 mmol) was added and stirring was continued for half an hour. After the reaction, acetic acid and water were added to quench the reaction, ethyl acetate was extracted, the organic phase was dried and concentrated, and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain 2,3-difluoro-6-isopropoxybenzaldehyde 182c (850 mg, colorless oily liquid), with a yield of 18.7%.
1 H NMR(400MHz,CDCl 3 )δ10.44–10.35(m,1H),7.29(dd,J＝13.0,5.3Hz,1H),6.71(ddd,J＝9.3,3.1,2.1Hz,1H),4.67–4.57(m,1H),1.39(d,J＝6.1Hz,6H).
The third step
Preparation of (2,3-difluoro-6-isopropoxyphenyl) methanol
2,3-difluoro-6-isopropoxybenzaldehyde 182c (850mg, 4.25mmol) was dissolved in methanol (10 mL), sodium borohydride (322mg, 8.5 mmol) was added at 0 ℃, the reaction mixture was stirred at that temperature for 20 minutes, after the completion of the reaction, a saturated aqueous solution of ammonium chloride was added and quenched, extracted with ethyl acetate, washed with an aqueous sodium chloride solution, and the organic phase was dried and concentrated, and then separated and purified with a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain (2,3-difluoro-6-isopropoxyphenyl) methanol 182d (680 mg, yellow oily liquid) in a yield of 79.1%.
1 H NMR(400MHz,CDCl 3 )δ7.02(dd,J＝19.0,9.1Hz,1H),6.59(ddd,J＝9.2,3.4,2.0Hz,1H),4.75(d,J＝1.4Hz,2H),4.57(dt,J＝12.2,6.1Hz,1H),2.55(s,1H),1.37(d,J＝6.1Hz,6H).
The fourth step
Preparation of 2- (bromomethyl) -3,4-difluoro-1-isopropoxybenzene
(2,3-difluoro-6-isopropoxyphenyl) methanol 182d (680mg, 3.36mmol) was dissolved in methylene chloride (20 mL), triphenylphosphine (1.06g, 4.03mmol) was added at 0 ℃ and stirred for 10 minutes, carbon tetrabromide (1.34g, 4.03mmol) was further added and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain 2- (bromomethyl) -3,4-difluoro-1-isopropoxybenzene 182e (600 mg, yellow solid) in 67.3% yield.
1 H NMR(400MHz,CDCl 3 )δ7.04(dd,J＝19.0,9.3Hz,1H),6.58(ddd,J＝9.2,3.5,2.1Hz,1H),4.60–4.53(m,3H),1.38(d,J＝6.1Hz,6H).
The fifth step
Preparation of N- (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline (382mg, 2.05mmol) is dissolved in DMF (5 mL), then 2- (bromomethyl) -3,4-difluoro-1-isopropoxybenzene 182e (600mg, 2.26mmol) and potassium carbonate (567mg, 4.1mmol) are added, the reaction solution reacts at 80 ℃ for 2 hours, after the reaction is finished, water is added into the reaction solution, ethyl acetate is extracted, and after drying and concentration, the reaction solution is separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to obtain N- (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 182f (340 mg, yellow solid) with the yield of 44.8%.
MS m/z(ESI)：371.1(M+1)。
The sixth step
N 1 Preparation of- (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-6-methoxyphenyl-1,3-diamine
Dissolving N- (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 182f (340mg, 0.918mmol) in ethanol/ammonium chloride saturated aqueous solution =4/1 (10 mL), adding iron powder (514mg, 9.18mmol), stirring the reaction solution at 80 ℃ for 2 hours, after the reaction is completed, suction-filtering the reaction solution, extracting the filtrate with ethyl acetate, drying and concentrating the organic phase, and separating with a silica gel column (petroleum ether/ethyl acetate = 10/1)Separating and purifying to obtain N 1 - (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-6-methoxyphenyl-1,3-diamine 182g (190 mg, yellow solid) in 60.8% yield.
MS m/z(ESI)：341.1(M+1)。
Seventh step
Preparation of dimethyl 4- (3- (5- ((2,3-difluoro-6-isopropylbenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
Will N 1 - (2,3-difluoro-6-isopropoxybenzyl) -4-fluoro-6-methoxyphenyl-1,3-diamine 182g (190mg, 0.558mmol) and 4- [ ((phenoxycarbonyl) amino]Thiophene-2,3-dicarboxylic acid methyl ester (225mg, 0.67mmol) was dissolved in tetrahydrofuran (10 mL), triethylamine (1699 mg, 1.67mmol) was added to the reaction solution, the reaction solution was stirred at room temperature for 16 hours, and after completion of the reaction, concentration was performed and purification was performed with silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4- (3- (5- ((2,3-difluoro-6-isopropylbenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 182h (270 mg, yellow solid) in 83.1% yield.
MS m/z(ESI)：582.1(M+1)。
Eighth step
Preparation of 3- (5- ((2,3-difluoro-6-isopropoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((2,3-difluoro-6-isopropylbenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 182h (270mg, 0.464mmol) was dissolved in tetrahydrofuran/water =1/1 (10 mL), lithium hydroxide (22mg, 0.8mmol) was added, the reaction solution was stirred at room temperature for 1 hour, and after completion of the reaction, the reaction solution was concentrated to give 3- (5- ((2,3-difluoro-6-isopropoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-101 (141 mg, yellow solid). The yield was 56.7%.
MS m/z(ESI)：536.0(M+1)。
HPLC:99.38％(214nm),97.78％(254nm)。
1 H NMR(400MHz,DMSO-d6)δ14.60(s,1H),11.99(s,1H),7.38(s,1H),7.28(dd,J＝19.6,9.5Hz,1H),6.97(d,J＝11.4Hz,1H),6.85(ddd,J＝9.0,3.2,1.4Hz,1H),6.78(d,J＝7.5Hz,1H),4.97(t,J＝6.3Hz,1H),4.66(dt,J＝12.0,6.0Hz,1H),4.22(d,J＝5.9Hz,2H),3.84(s,3H),1.25(dd,J＝6.0,3.6Hz,6H).
Example 97
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001841
First step of
Preparation of 4- (cyclopropylmethoxy) -1,2-difluorobenzene
3,4-difluorophenol 184a (1.30g, 10.00mmol) was dissolved in N, N-dimethylformamide (5 mL), bromomethylcyclopropane (1.35g, 10.00mmol), potassium carbonate (4.15g, 30.00mmol) and potassium iodide (1.66g, 10.00mmol) were added in this order, and the reaction mixture was reacted at 90 ℃ for 16 hours. After the reaction was complete, water (10 mL) was added, extracted with ethyl acetate (3X 10 mL), and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/1) to give 4- (cyclopropylmethoxy) -1,2-difluorobenzene 184b (820 mg, colorless oily liquid) in 40% yield.
1 H NMR(400MHz,CDCl 3 )δ7.09–6.99(m,1H),6.73–6.68(m,1H),6.62–6.56(m,1H),3.74(d,J＝6.8Hz,2H),1.32–1.18(m,1H),0.72–0.56(m,2H),0.44–0.27(m,2H).
Second step of
Preparation of 6- (cyclopropylmethoxy) -2,3-difluorobenzaldehyde
4- (Cyclopropylmethoxy) -1,2-difluorobenzene 184b (690mg, 3.75mmol) was dissolved in ultra-dry tetrahydrofuran (10.0 mL). After the reaction flask was stirred at-78 ℃ for 0.5 hour, a tetrahydrofuran solution of lithium diisopropylamide (3.7 mL, 2mol/L) was slowly added over 15 minutes, the reaction mixture was stirred at-78 ℃ for 20 minutes, N-dimethylformamide (301mg, 4.12mmol) was slowly added dropwise, the mixture was further reacted at-78 ℃ for 0.5 hour, and after completion of the reaction, room temperature was recovered, acetic acid (0.2 mL), water (1.5 mL) and ethyl acetate (3X 10 mL) were slowly added dropwise, followed by extraction, washing with water (10 mL) and saturated brine (10 mL) in this order, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 95/5) to give 6- (cyclopropylmethoxy) -2,3-difluorobenzaldehyde c (623 mg, colorless oily liquid), yield: 71%.
MS m/z(ESI)：213.1(M+1).
1 H NMR(400MHz,CDCl 3 )δ10.34(s,1H),7.30–7.18(m,1H),6.64–6.61(m,1H),3.84(d,J＝6.8Hz,2H),1.23–1.19(m,1H),0.64–0.52(m,2H),0.37–0.21(m,2H).
The third step
Preparation of (6- (cyclopropylmethoxy) -2,3-difluorophenyl) methanol
6- (Cyclopropylmethoxy) -2,3-difluorobenzaldehyde 184c (623mg, 2.94mmol) was dissolved in methanol (10 mL) and sodium borohydride (222mg, 5.87mmol) was added under an ice-water bath. The reaction mixture was reacted at 0 ℃ for 20 minutes. After the reaction was completed, saturated ammonium chloride was added to quench, and then diluted with ethyl acetate (15 mL), washed with water (5 mL) and saturated brine (2 × 5 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate = 100/0) to give (6- (cyclopropylmethoxy) -2,3-difluorophenyl) methanol 184d (542 mg, white solid) in 78% yield.
MS m/z(ESI)：197.2(M-18).
1 H NMR(400MHz,CDCl 3 )δ7.07–6.94(m,1H),6.56–6.52(m,1H),4.80(d,J＝2.0Hz,2H),3.84(d,J＝7.2Hz,2H),2.12(s,1H),1.34–1.20(m,1H),0.74–0.60(m,2H),0.44–0.25(m,2H).
The fourth step
Preparation of 2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene
(6- (cyclopropylmethoxy) -2,3-difluorophenyl) methanol 184d (442mg, 2.06mmol) was dissolved in dichloromethane (4 mL), triphenylphosphine (649mg, 2.48mmol) and carbon tetrabromide (821mg, 2.48mmol) were added in this order in an ice-water bath, and the reaction mixture was reacted at 0 ℃ for 1 hour. After the reaction was complete, water (2 mL) was added, extracted with dichloromethane (3X 10 mL), and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 90/10) to give 2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene 184e (400 mg, white solid) in 63% yield.
1 H NMR(400MHz,CDCl 3 )δ7.12–6.97(m,1H),6.62–6.48(m,1H),4.60(d,J＝1.6Hz,1H),3.87(t,J＝5.6Hz,2H),1.36–1.22(m,1H),0.71–0.61(m,2H),0.45–0.32(m,2H).
The fifth step
Preparation of N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene 184e (400mg, 1.44mmol) was dissolved in acetonitrile (10 mL), 4-fluoro-2-methoxy-5-nitroaniline (322mg, 1.73mmol), potassium carbonate (599 mg, 4.33mmol) and potassium iodide (240mg, 1.44mmol) were added sequentially, and the reaction was reacted at 65 ℃ for 2 hours. After the reaction was complete, water (2 mL) was added, extracted with ethyl acetate (3X 10 mL), and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 75/25) to give N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 184f (520 mg, yellow solid) in 85% yield.
MS m/z(ESI)：383.1(M+1).
1 H NMR(400MHz,CDCl 3 )δ7.68(d,J＝7.2Hz,1H),7.04(q,J＝9.2Hz,1H),6.63(d,J＝12.2Hz,1H),6.60–6.53(m,1H),4.57(s,2H),3.95(s,3H),3.90(d,J＝6.8Hz,2H),1.47–1.27(m,1H),0.73–0.65(m,2H),0.41(q,J＝4.8Hz,2H).
The sixth step
N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine
N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 184f (100mg, 0.26mmol) was dissolved in tetrahydrofuran/H 2 O (2.5mL, V/V = 4:1). Then, ammonium chloride (42mg, 0.78mmol) and iron powder (44mg, 0.78mmol) were added, and the reaction mixture was reacted at 25 ℃ for 16 hours. Then filtering to obtain filtrateExtract with dichloromethane (2 × 10 mL). The organic phases are combined, dried by anhydrous sodium sulfate and concentrated to obtain N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 184g (95 mg, brown solid) in 93% yield.
MS m/z(ESI)：353.1(M+1).
Seventh step
Preparation of dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylate
Will N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 184g (95mg, 0.27mmol) was dissolved in tetrahydrofuran (2 mL) with dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylate (90mg, 0.27mmol). Then, triethylamine (82mg, 0.81mmol) was added, and the reaction mixture was reacted at 25 ℃ for 2 hours. After the reaction was complete, water (2 mL) was added, extracted with ethyl acetate (3X 10 mL), and washed with saturated brine (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 75/25) to give 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylic acid dimethyl ester 184h (107 mg, brown oily liquid) in 93% yield.
MS m/z(ESI)：594.1(M+1).
Eighth step
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2,3-dicarboxylate 184h (107mg, 0.18mmol) was dissolved in tetrahydrofuran with lithium hydroxide (13mg, 0.54mmol): water =4:1 (2.5 mL). The reaction mixture was reacted at room temperature for 2 hours. After the reaction was complete, the reaction was directly filtered, concentrated and purified by preparative reverse-phase column (water: acetonitrile = 20).
MS m/z(ESI)：548.1(M+1).
HPLC:100.00％(214nm),100.00％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.59(s,1H),11.98(s,1H),7.37(s,1H),7.33–7.25(m,1H),6.97(d,J＝11.6Hz,1H),6.84–6.75(m,2H),4.97(t,J＝6.4Hz,1H),4.27(d,J＝6.0Hz,1H),3.88–3.87(m,1H),3.84(s,3H),1.26–1.13(m,1H),0.54–0.47(m,2H),0.33–0.25(m,2H).
19 F NMR(376MHz,d 6 -DMSO)δ-134.41,-140.42,-148.22.
Example 98
3- (5- ((2,3-difluoro-6-methoxyphenyl) methoxy) -2-fluoro-4-hydroxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001861
First step of
Preparation of 4-fluoro-2-methoxy-5-nitrophenol 188b
To 4-fluoro-2-methoxy-5-nitrophenol 188a (2g, 0.0107 mol) were added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120 ℃ for 12 hours. After completion of the reaction, it was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/80% petroleum ether) to give 4-fluoro-2-methoxy-5-nitrophenol (1.1 g, yellow solid) with yield: and 59.81 percent.
MS m/z(ESI)：174.1(M+1).
Second step of
Preparation of 2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 188c
4-fluoro-2-methoxy-5-nitrophenol 188b (100mg, 0.5777mmol) is dissolved in 20mL DMF and NaH (27.73mg, 1.1554mmol) is added at 0 ℃ and stirred at this temperature for 10 minutes, followed by addition of 2-bromo-3,4-difluoro-1-methoxybenzene (128.84mg, 0.5777mmol) and stirring for 10 minutes and then reaction at 25 ℃ for 1 hour. After completion of the reaction, the reaction system was made acidic with 2N diluted hydrochloric acid, and extracted with ethyl acetate (2X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: 10% ethyl acetate/90% petroleum ether) to give 2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 188c (100 mg, yellow solid), yield: 43.93 percent.
MS m/z(ESI)：330.1(M+1)。
The third step
Preparation of 4-amino-2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluorophenol 188d
2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 188c (100mg, 0.30mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (169.62mg, 3.04mmol) were added, and the reaction mixture was reacted at 25 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (2 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 4-amino-2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluorophenol 188d (80 mg, yellow solid), yield: 79.22 percent.
MS m/z(ESI)：300.1(M+1)。
The fourth step
Preparation of 4- [ ((5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester 188e
4-amino-2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluorophenol (80mg, 0.25mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester (158mg, 0.50mmol) were dissolved in tetrahydrofuran (5 mL). Then, triethylamine (77mg, 0.75mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 1 hour. The reaction solution was not subjected to any post-treatment to give crude 4- [ ((5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester 188e.
MS m/z(ESI)：541.1(M+1)。
The fifth step
Preparation of 3- (5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ((5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester 188e (5 mL stock solution) and lithium hydroxide (13mg, 0.53mmol) were dissolved in methanol: water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 2 hours. After the reaction was completed, the reaction mixture was directly concentrated and purified by reverse column (water: acetonitrile = 20) to give 3- (5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-103 (80 mg, yellow solid) in 87.46% yield.
MS m/z(ESI)：495.1(M+1)。
1 H NMR(400MHz,DMSO-d6)δ14.59(s,1H),11.94(s,1H),9.95(s,1H),7.47(dd,J＝19.6,9.6Hz,1H),7.33(s,1H),7.16(d,J＝7.4Hz,1H),6.92-6.89(m,1H),6.80(d,J＝11.0Hz,1H),4.96(s,2H),3.81(s,3H).
Example 99
3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene (3,4-d) pyrimidine-5-carboxylic acid
Figure BDA0003608368340001871
First step of
Preparation of N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline
N- [ (6-ethoxy-2,3-difluorophenyl) methyl ] -4-fluoro-2-methoxy-5-nitroaniline (2.5g, 7 mmol), formaldehyde (0.45g, 14mmol) were dissolved in acetonitrile (30 mL) and stirred at 40 ℃ under nitrogen for 0.5h. Sodium cyanoborohydride (0.66g, 10.5 mmol) and acetic acid (0.02g, 0.3mmol) were added. The reaction mixture was stirred at 40 ℃ for 4h. After the reaction was complete, 1Ml of water was added and quenched, concentrated, diluted with water (30 Ml), extracted with EA (20 Ml), the organic phase was concentrated and the crude product was purified by silica gel chromatography to give N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 189b (800 mg, yellow solid) yield: 27.1 percent.
MS m/z(ESI)：371.1(M+H)。
Second step of
N 1 - (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxyPreparation of Yl-N1-toluene-1,3-diamine
N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline (700mg, 1.89mmol), ammonium chloride (553mg, 9.45mmol) was dissolved in EtOH/H2O =4:1 (10 mL) and iron (529mg, 9.45mmol) was added. The reaction mixture was stirred at 80 ℃ for 2 hours. After the reaction, filtering, concentrating the filtrate, and purifying the obtained crude product by silica gel chromatography to obtain N 1 - (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxy-N1-toluene-1,3-diamine (400 mg, yellow solid), yield: 57.21 percent.
MS m/z(ESI)：341.2(M+H)。
The third step
Preparation of dimethyl 4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
Will N 1 - (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxy-N1-toluene-1,3-diamine (400mg, 1.175mmol), 4- ((phenoxycarbonyl) amino) thiophene-2,3 dimethyl dicarboxylate (394mg, 1.175mmol) were dissolved in THF (20 mL) and triethylamine (712mg, 7.052mmol) was added dropwise. The reaction mixture was stirred at 25 ℃ for 1 hour. After the reaction was completed, the reaction mixture was concentrated to give a crude product, which was purified by a silica gel column (dichloromethane: methanol =10: 1) to give 4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester (500 mg, yellow solid), yield: 68.7 percent.
MS m/z(ESI)：582.1(M+H)。
The fourth step
Preparation of 3- (5- ((6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene (3,4-d) pyrimidine-5-carboxylic acid
4- (3- (5- ((6-ethoxy-2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester (500mg, 0.26mmol) lithium hydroxide monohydrate (22mg 0.53mmol) was dissolved in methanol: tetrahydrofuran: water =1:1;1 (6 mL). The reaction mixture was stirred at 25 ℃ for 2 hours. After the reaction was finished, concentrated and the crude product was purified by silica gel chromatography (dichloromethane: methanol = 10): 8.8 percent.
MS m/z(ESI)：536.1(M+H).
1 H NMR(400MHz,DMSO-d 6 ))δ14.68(s,1H),11.91(s,1H),7.37(s,1H),7.24(dd,J＝19.4,9.5Hz,1H),7.04(t,J＝18.3Hz,1H),6.79(d,J＝8.0Hz,1H),6.72(dd,J＝9.2,2.0Hz,1H),4.36(q,J＝14.2Hz,2H),3.91(s,3H),3.88–3.71(m,2H),2.52(s,3H),1.14(t,J＝6.9Hz,3H).
Example 100
3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001891
First step of
Preparation of N-cyclopropyl-4-fluoro-2-methoxy-5-nitroaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 190a (1g, 0.004mol) was dissolved in toluene (20 mL), cyclopropylamine (0.27g, 0.0048mol), tris-dibenzylideneacetone dipalladium (0.37g, 0.0004mol), 2- (dicyclohexylphosphine) -3,6-dimethoxy-2 '-4' -6 '-tri-I-propyl-11' -biphenyl (0.64g, 0.0012mol), sodium tert-butoxide (0.58g, 0.0062mol) were added, and the reaction was carried out at 100 ℃ for 16 hours under nitrogen protection, after completion of the reaction, the temperature was returned to room temperature, and filtered. Water (10 mL) was added, extracted with ethyl acetate (3 × 10 mL), the organic phase was dried, concentrated, and the concentrate was further purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to give N-cyclopropyl-4-fluoro-2-methoxy-5-nitroaniline 190b (400 mg, white solid). Yield: and 43 percent.
MS m/z(ESI)：227(M+1)。
Second step of
Preparation of N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
N-cyclopropyl-4-fluoro-2-methoxy-5-nitroaniline 190b (400mg, 1.5932mmol) was dissolved in acetonitrile (10 mL), and 2- (bromomethyl) -1-ethoxy-3,4-difluorobenzene (396.43mg, 1.7525mmol), potassium carbonate (660.57mg, 4.7796mmol), potassium iodide (26.45mg, 0.1593mmol) were added. After the reaction was completed, acetonitrile was removed by rotation, water (10 mL) and ethyl acetate (3X 10 mL) were added for extraction, and the organic phase was dried and concentrated. The concentrate was further purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to give N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 190c (300 mg, yellow solid). Yield: 45 percent.
MS m/z(ESI)：397(M+1)。
The third step
Preparation of N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine
Dissolving N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 190c (300mg, 0.7569mmol) in acetonitrile (5 mL), adding iron powder (211.36mg, 3.7845mmol) and ammonium chloride (121.46mg, 2.2707mmol), reacting at 80 ℃ for one hour, removing acetonitrile by spinning off after the reaction is finished, adding water (10 mL) and ethyl acetate (3 x10 mL), extracting, drying and concentrating the organic phase. The concentrate was further purified by silica gel column (petroleum ether/ethyl acetate = 5/1) to give N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 190d (220 mg, yellow solid). Yield: 75 percent.
MS m/z(ESI)：367(M+1)。
The fourth step
Preparation of 4- (3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
N-cyclopropyl-N- (6-ethoxy-2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 190d (220mg, 0.6005mmol) was dissolved in tetrahydrofuran (5 mL), and 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (241.64mg, 0.0672mmol), triethylamine (182.29mg, 1.8015mmol) were added. After the reaction was completed, water (10 mL) and ethyl acetate (3X 10 mL) were added for extraction. The organic phase was dried and concentrated. The concentrate was further purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to give 4- (3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 190e (150 mg, yellow solid). Yield: 40 percent.
MS m/z(ESI)：607(M+1)
The fifth step
Preparation of 3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 190e (150mg, 0.3456mmol) was dissolved in a solution of methanol/tetrahydrofuran/water = 1/1/1/1 (3 mL), lithium hydroxide (41.39mg, 1.728mmol) was added and reacted at room temperature for one hour, after completion of the reaction, the solvent was spun dry and the crude product was prepared by preparative column (acetonitrile/water = 1/4) to give 3- (5- (cyclopropyl (6-ethoxy-2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-3562 zft 1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-105 (61 mg, yellow solid). Yield: 31 percent.
MS m/z(ESI)：562(M+1)
HPLC:100％(214nm),99.70％(254nm).
1 H NMR(400MHz,d 6 -DMSO)δ14.68(s,1H),11.66(s,1H),7.39–7.25(m,1H),7.13(s,1H),6.97(s,1H),6.86(d,J＝7.8Hz,1H),6.67(d,J＝7.2Hz,1H),4.47(dd,J＝25.1,13.9Hz,2H),3.83(d,J＝123.0Hz,6H),1.05(s,3H),0.49(s,2H),0.33(d,J＝17.3Hz,2H).
Example 101
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001901
First step of
Preparation of N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline (1g, 5.37mmol) was dissolved in methanol (20 mL), several drops of acetic acid and 6- (cyclopropylmethoxy) -2,3-difluorobenzaldehyde 191a (1.37g, 6.45mmol) were added, the reaction was stirred at room temperature for 15 minutes, followed by addition of sodium cyanoborohydride (1.01g, 16.11mmol), the reaction was stirred at 40 ℃ for 2 hours, after completion of the reaction, quenching was performed by addition of an aqueous ammonium chloride solution, extraction was performed with ethyl acetate, organic phase was dried and concentrated, and then N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 191b (1 g, yellow solid) was isolated and purified by silica gel column (petroleum ether/ethyl acetate = 3/1) to give a yield of 48.7%.
MS m/z(ESI)：383.1(M+1)。
Second step of
Preparation of N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline
Dissolving N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 191b (1g, 2.61mmol) in methanol (20 mL), adding a few drops of acetic acid and aqueous formaldehyde, stirring the reaction mixture at room temperature for 15 minutes, adding sodium cyanoborohydride (493mg, 7.85mmol), stirring the reaction mixture at 40 ℃ for 16 hours, after the reaction is finished, adding aqueous ammonium chloride, quenching, extracting with ethyl acetate, concentrating by organic drying, and separating and purifying with a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 191c (900 mg, yellow solid) with a yield of 87%.
MS m/z(ESI)：397.1(M+1)。
The third step
N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxy-N 1 Preparation of (E) -methylbenzene-1,3-diamine
Dissolving N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 191c (900mg, 2.27mmol) in ethanol/ammonium chloride aqueous solution =4/1 (25 mL), adding iron powder (1.27g, 22.7 mmol), stirring the reaction solution at 80 ℃ for 2 hours, filtering under suction after the reaction is finished, concentrating the filtrate, and separating and purifying with a silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain N 1 - (6- (cyclopropylmethoxy) -2,3-difluoroBenzyl) -4-fluoro-6-methoxy-N 1 -methylbenzene-1,3-diamine 191d (430 mg, yellow solid) in 51.7% yield.
MS m/z(ESI)：367.2(M+1)。
The fourth step
Preparation of dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
Will N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxy-N 1 -methylbenzene-1,3-diamine 191d (430mg, 1.17mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (472mg, 1.41mmol) and triethylamine (356mg, 3.52mmol), the reaction solution was stirred at room temperature for 16 hours, and after completion of the reaction, the reaction solution was concentrated and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 191e (600 mg, yellow solid) with a yield of 84.4%.
MS m/z(ESI)：608.2(M+1)。
The fifth step
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 191e (600mg, 0.987mmol) was dissolved in tetrahydrofuran/water =1/1 (10 mL) and lithium hydroxide (71mg, 2.96mmol) was added, the reaction solution was stirred at room temperature for 1 hour, and after completion of the reaction, the reaction solution was concentrated to give 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-106 (194 mg, yellow solid) in 35% yield.
MS m/z(ESI)：562.1(M+1)。
HPLC:99.61％(214nm),98.94％(254nm)。
1 H NMR(400MHz,DMSO-d6)δ14.66(s,1H),11.90(s,1H),7.36(s,1H),7.22(dd,J＝19.3,9.5Hz,1H),7.05(d,J＝11.9Hz,1H),6.82(d,J＝8.0Hz,1H),6.73–6.68(m,1H),4.36(q,J＝13.7Hz,2H),3.90(s,3H),3.66(d,J＝6.9Hz,2H),2.54(s,3H),1.05(td,J＝7.4,3.8Hz,1H),0.55–0.47(m,2H),0.241–0.204(m,2H).
Example 102
3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001921
First step of
Preparation of N-cyclopropyl-N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
N-cyclopropyl-4-fluoro-2-methoxy-5-nitroaniline 192a (650mg, 2.87mmol) and 2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene (876mg, 3.16mmol) were dissolved in acetonitrile (30 mL), followed by the addition of potassium carbonate (792mg, 5.74mmol) and potassium iodide (476mg, 2.87mmol), and the reaction was stirred at 70 ℃ for 16 hours. After the reaction, water was added to the reaction solution, and ethyl acetate was extracted, and the organic phase was concentrated by drying and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to obtain N-cyclopropyl-N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 192b (480 mg, yellow solid) with a yield of 39.6%.
MS m/z(ESI)：423.1(M+1)。
Second step of
N 1 -cyclopropyl-N 1 Preparation of- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine
N-cyclopropyl-N- (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 192b (480mg, 1.14mmol) was dissolved in ethanol/ammonium chloride aqueous solution =4/1 (25 mL), followed by addition of iron powder (635mg, 11.4mmol), stirring of the reaction solution at 80 ℃ for 2 hours, completion of the reaction, filtration of the reaction solution, concentration of the filtrate, and silica gel column (petroleum jelly) chromatography Ether/ethyl acetate = 5/1) separation and purification to yield N 1 -cyclopropyl-N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 192c (180 mg, yellow solid) in 40.2% yield.
MS m/z(ESI)：383.1(M+1)。
The third step
Preparation of dimethyl 4- (3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
N is to be 1 -cyclopropyl-N 1 - (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1,3-diamine 192c (180mg, 0.459mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (1699 mg, 0.504mmol) and triethylamine (139mg, 1.37mmol), the reaction was stirred at 40 ℃ for 16 hours, and after completion of the reaction, the reaction was concentrated and purified by silica gel column (petroleum ether/ethyl acetate = 3/1) separation to give 4- (3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 192d (270 mg, yellow solid) in 92.8% yield.
MS m/z(ESI)：634.2(M+1)。
The fourth step
Preparation of 3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 192d (270mg, 0.426 mmol) was dissolved in tetrahydrofuran/water =1/1 (10 mL), followed by addition of lithium hydroxide (30mg, 1.28mmol), and the reaction solution was stirred at room temperature for 1 hour, after completion of the reaction, the reaction solution was concentrated to give 3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2,3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-107 (70 mg, yellow solid) which was prepared. Yield: 30 percent.
MS m/z(ESI)：588.1(M+1)。
HPLC:99.65％(214nm),98.94％(254nm)。
1H NMR(400MHz,DMSO)δ7.17(q,J＝9.6Hz,2H),6.99(d,J＝11.9Hz,1H),6.85(d,J＝8.1Hz,1H),6.64(d,J＝9.1Hz,1H),4.55–4.44(m,2H),3.87(s,3H),3.59(d,J＝7.0Hz,2H),2.34(s,1H),1.08–0.97(m,1H),0.58–0.48(m,4H),0.38–0.26(m,2H),0.20(q,J＝4.8Hz,2H).
Example 103
5- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
Figure BDA0003608368340001931
First step of
Preparation of 2,3-dicyano diethyl fumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL, 0.07mol) and ethyl 2-cyanoacetate 193a (5 mL, 0.04mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction mixture was refluxed and stirred at 90 ℃ for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid precipitated. The reaction was filtered and washed with glacial ethanol to give 2,3-diethyl dicyano fumarate 193b (2.3 g, white solid) in yield: 21.04%.
MS m/z(ESI)：223.1(M+1).
Second step of
Preparation of 5-amino-1H-pyrazole-3,4-dicarboxylic acid diethyl ester
2,3-dicyano diethyl fumarate 193b (2g, 0.009mol) was dissolved in ethanol (10 mL), and acethydrazide (0.87g, 0.0099mol) and sodium acetate (0.07g, 0.0009mol) were added to react the reaction mixture at 90 ℃ for 0.5 hour. After completion of the reaction, it was cooled to room temperature and extracted with dichloromethane (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 50% ethyl acetate/50% petroleum ether) to give 5-amino-1H-pyrazole-3,4-dicarboxylic acid diethyl ester 193c (1.2 g, white solid), yield: 58.89%.
MS m/z(ESI)：228.1(M+1).
The third step
Preparation of diethyl 5- ([ (5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3,4-dicarboxylate
5-amino-1H-pyrazole-3,4-dicarboxylic acid diethyl ester 193c (501.6mg, 2.21mmol) was dissolved in dichloromethane (20 mL), 1,1' -carbonyldiimidazole (429.54mg, 2.65mmol), triethylamine (670.13mg, 6.62mmol) and 5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyaniline (780mg, 2.21mmol) were added, and the reaction solution was reacted at 25 ℃ for 1.5 hours. After the reaction was completed, water quenching was added, extraction was performed with dichloromethane (3 × 30 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 5% methanol/95% dichloromethane) to give 5- ([ (5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3,4-dicarboxylic acid diethyl ester 193d (500 mg, white solid), yield: 37.3 percent.
MS m/z(ESI)：607.2(M+1).
The fourth step
Preparation of 5- (5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-1H, 7H-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
5- ([ (5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3,4-dicarboxylic acid diethyl ester 193d (100mg, 0.18mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (38mg, 0.90mmol) was added and the reaction stirred at 25 ℃ for 4H. After completion of the reaction, the reaction system was made acidic with 2N diluted hydrochloric acid and extracted with dichloromethane (2X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 10% ethyl acetate/80% petroleum ether) to give 5- (5- ([ 6- (cyclopropylmethoxy) -2,3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) -4,6-dioxo-1h, 7 h-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid Cpd-108 (3.5 mg, white solid), yield: 3.82 percent.
MS m/z(ESI)：533.1(M+1).
HPLC:96.53％(214nm),97.43％(254nm).
1 H NMR(400MHz,DMSO-d6)δ7.44(d,J＝9.6Hz,1H),7.09(s,1H),6.89(d,J＝8.8Hz,2H),5.01(s,2H),3.87(d,J＝7.0Hz,2H),3.80(s,3H),1.24-1.16(m,1H),0.48(d,J＝6.4Hz,2H),0.26(d,J＝5.8Hz,2H).
Example 104
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thiophene [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340001941
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thiophene [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Oxalyl chloride (200mg, 1.57mmol) and one drop of N, N-dimethylformamide were added dropwise to a solution of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxy-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid 177a (400mg, 0.79mmol) in dichloromethane (5 mL), and the reaction was stirred at 25 ℃ for 15 minutes. After the reaction was completed, the reaction solution was concentrated, dissolved by adding tetrahydrofuran (3 mL), and then added dropwise to an aqueous solution dissolved with sodium borohydride (60mg, 1.57mmol), after the reaction was stirred at 0 ℃ for 15 minutes, water and ethyl acetate (3 × 10 mL) were added for extraction, the organic phase was dried with sodium sulfate hydrate, concentrated, and the concentrate was purified by preparative column to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thiophene [3,4-d ] pyrimidine-2,4 (1h, 3h) -dione Cpd-109 (202.62 mg, white solid) with yield: 52 percent.
MS m/z(ESI)：495.0(M+1)。
HPLC:100％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO)δ11.34(s,1H),7.48(dd,J＝19.7,9.5Hz,1H),7.18(d,J＝7.4Hz,1H),7.06(d,J＝11.5Hz,1H),6.93-6.89(m,1H),6.72(s,1H),5.97(t,J＝5.5Hz,1H),5.06–4.99(m,2H),4.95(s,2H),3.81(s,3H),3.78(s,3H).
Example 105
3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001951
First step of
Preparation of tert-butyl (tert-butoxycarbonyl) (2-chloro-5-fluoropyridin-4-yl) carbamate
After the reaction solution was stirred at room temperature for 16 hours by dissolving 2-chloro-4-amino-5-fluoropyridine 196a (1g, 6.82mmol) in acetonitrile (20 mL), followed by addition of di-tert-butyl dicarbonate (3.72g, 17.05mmol) and 4-dimethylaminopyridine (167mg, 1.36mmol), the reaction solution was concentrated and then purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to give tert-butyl (tert-butoxycarbonyl) (2-chloro-5-fluoropyridin-4-yl) carbamate 196b (1 g, white solid) in a yield of 42.3%.
MS m/z(ESI)：347.1(M+1)。
Second step of
Preparation of tert-butyl (tert-butoxycarbonyl) (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) carbamate
Tert-butyl (tert-butoxycarbonyl) (2-chloro-5-fluoropyridin-4-yl) carbamate 196b (1g, 2.88mmol) and (6- (cyclopropylmethoxy) -2,3-difluorophenyl) methanol (617mg, 2.88mmol) were dissolved in 1,4-dioxane (20 mL) and Brettphos (309mg, 0.576 mmol), pd, and 2 (dba) 3 (264mg, 0.288mmol) and cesium carbonate (1.88g, 5.76mmol), the reaction mixture was stirred at 80 ℃ for 2 hours, after completion of the reaction, the reaction mixture was washed with water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to give tert-butyl (tert-butoxycarbonyl) (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) carbamate 196c (400 mg, yellow solid) in 26.5% yield.
MS m/z(ESI)：525.2(M+1)。
The third step
Preparation of 2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-amine
Tert-butyl (tert-butoxycarbonyl) (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) carbamate 196c (400mg, 0.762mmol) was dissolved in dichloromethane (10 mL), followed by addition of HCl/1,4-dioxane solution (5 mL), the reaction was stirred at 35 ℃ for 16 hours, after completion of the reaction, the reaction was concentrated, a saturated aqueous sodium bicarbonate solution was added, ethyl acetate was extracted, the organic phase was dried over anhydrous sodium sulfate, and after concentration, it was separated and purified by a silica gel column (petroleum ether/ethyl acetate = 2/1) to give 2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-amine 196d (200 mg, yellow oily liquid) in 80.9% yield.
MS m/z(ESI)：325.1(M+1)。
The fourth step
Preparation of dimethyl 4- (3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) ureido) thiophene-2,3-dicarboxylate
2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-amine 196d (200mg, 0.617mmol) was dissolved in tetrahydrofuran (5 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (621mg, 1.851mmol) and triethylamine (187mg, 1.851mmol), the reaction was stirred at room temperature for 16 hours, after completion of the reaction, the reaction was concentrated and then purified by silica gel column (petroleum ether/ethyl acetate = 2/1) to give 4- (3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 196e (10 mg, yellow solid) in 2.9% yield.
MS m/z(ESI)：566.2(M+1)。
The fifth step
Preparation of 3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 196e (10mg, 0.017mmol) was dissolved in tetrahydrofuran/water =1/1 (2 mL), followed by addition of lithium hydroxide (2mg, 0.085mmol), the reaction mixture was stirred at room temperature for 1 hour, and after completion of the reaction, 3- (2- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-110 (2 mg, yellow solid) was prepared by spin-drying, with a yield of 22.6%.
MS m/z(ESI)：520.0(M+1)。
HPLC:92.11％(214nm),66.51％(254nm)。
1 H NMR(400MHz,dmso)δ7.71(s,1H),7.38(dd,J＝19.8,9.5Hz,1H),7.03(d,J＝4.5Hz,1H),6.84(d,J＝8.1Hz,1H),6.67(s,1H),5.35–5.22(m,2H),3.84(d,J＝6.8Hz,2H),0.80(t,J＝6.3Hz,1H),0.51–0.41(m,2H),0.23(dt,J＝10.3,5.3Hz,2H).
Example 106
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340001961
First step of
Preparation of methyl (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidin-5-yl) methanesulfonate
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione Cpd-117 (14mg, 0.026mmol) was dissolved in dichloromethane (1 mL), DIPEA (10mg, 0.078mmol) was added, msCl (3mg, 0.026mmol) was added at 0 ℃, and the reaction was stirred for 1 hour, followed by direct use in the next step without any treatment.
MS m/z(ESI)：549.1(M+1)。
Second step of
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Dimethylamine hydrochloride (4 mg, 0.052mmol) was added to the reaction solution in the previous step, the reaction solution was stirred at room temperature for 1 hour, and after completion of the reaction, the reaction solution was concentrated to give methyl (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidin-5-yl) methanesulfonate, cpd-111 (2.6 mg, white solid), in 18% yield.
MS m/z(ESI)：562.1(M+1)。
HPLC:100％(214nm),100％(254nm)。
1 H NMR(400MHz,DMSO)δ11.37(s,1H),7.43(dd,J＝19.7,9.5Hz,1H),7.19(d,J＝7.4Hz,1H),7.07(d,J＝11.5Hz,1H),6.88(ddd,J＝9.2,3.4,1.6Hz,1H),6.78(s,1H),5.00(s,2H),4.10(s,2H),3.87(d,J＝6.9Hz,2H),3.80(s,3H),2.33(d,J＝1.7Hz,6H),1.20–1.10(m,1H),0.51–0.44(m,2H),0.29–0.23(m,2H).
Example 107
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [2,3-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340001971
First step of
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [2,3-d ] pyrimidine-5-carbaldehyde
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [2,3-d ] pyrimidine-2,4 (1H, 3H) -dione 198a (40mg, 0.075mmol) and manganese dioxide (33mg, 0.37mmol) were dissolved in dichloromethane (5 mL). The reaction solution was stirred at 25 ℃ for 16 hours. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [2,3-d ] pyrimidine-5-carbaldehyde 198b (35 mg, yellow solid), yield: 77% of the reaction solution was used directly in the next step.
MS m/z(ESI)：533.0(M+1)。
Second step of
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [2,3-d ] pyrimidine-2,4 (1H, 3H) -dione
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [2,3-d ] pyrimidine-5-carboxaldehyde 198b (35mg, 0.066 mmol) and dimethylamino hydrochloride (11mg, 0.131mmol) were dissolved in methanol (5 mL). After the reaction was stirred at 40 ℃ for 30 minutes, sodium cyanoborohydride (21mg, 0.328mmol) was added to the reaction solution, and the reaction solution was stirred at 40 ℃ for 1 hour. After the reaction was complete, the crude product was purified by preparative purification to give 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [2,3-d ] pyrimidine-2,4 (1h, 3h) -dione Cpd-112 (9.26 mg, white solid), yield: 22.53 percent.
MS m/z(ESI)：562.0(M+1)。
HPLC:90.19％(214nm),95.83％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.42(dd,J＝19.6,9.6Hz,1H),7.01(dd,J＝15.6,9.6Hz,2H),6.90–6.83(m,1H),6.58(d,J＝16.8Hz,1H),5.00(s,2H),3.86(d,J＝7.0Hz,2H),3.77(s,3H),3.73(s,2H),2.32(s,6H),1.15-1.10(m,1H),0.50–0.42(m,2H),0.27–0.19(m,2H).)
Example 108
3- (2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxolan-4-yl) methoxy ] -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340001981
First step of
Preparation of 6-fluoro-2,3-dimethoxybenzoic acid 199b
LDA (1.51g, 0.014mol) was added dropwise to 4-fluoro-1,2-dimethoxybenzene 199a (2g, 0.013mol) under nitrogen atmosphere at-78 deg.C, and stirred at-60 deg.C for 30 minutes. Then dry carbon dioxide was added and stirring was continued for 60 minutes. After the reaction was completed, the temperature was slowly returned to room temperature and water was slowly added, the reaction system was adjusted to acidity (pH =3 to 4) with 2N diluted hydrochloric acid, and ethyl acetate was added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to give 6-fluoro-2,3-dimethoxybenzoic acid 199b (1.2 g, yellow liquid), yield: 42.19 percent.
MS m/z(ESI)：201.1(M+1).
Second step of
Preparation of 6-fluoro-2,3-dihydroxybenzoic acid 199c
To 6-fluoro-2,3-dimethoxybenzoic acid 199b (1.2g, 0.0107 mol) were added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120 ℃ for 12h. After completion of the reaction, it was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/80% petroleum ether) to give 6-fluoro-2,3-dihydroxybenzoic acid 199c (0.8 g, yellow liquid), yield: 70.00 percent.
MS m/z(ESI)：173.1(M+1).
The third step
Preparation of 6-fluoro-2,3-dihydroxy methyl benzoate 199d
6-fluoro-2,3-dihydroxybenzoic acid 199c (0.8g, 0.0046mol) was dissolved in 20mL of methanol followed by the addition of SOCl at 0 deg.C 2 (1.64g, 0.0138mmol), and the mixture solution was stirred at 70 ℃ for 16h. After completion of the reaction, it was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 10% ethyl acetate/90% petroleum ether) to give methyl 6-fluoro-2,3-dihydroxybenzoate 199d (0.6 g, yellow liquid), yield: 63.04%.
MS m/z(ESI)：187.1(M+1).
The fourth step
Preparation of methyl 5-fluoro-2H-1,3-benzodioxolane-4-carboxylate 199e
Methyl 6-fluoro-2,3-dihydroxybenzoate 199d (500mg, 2.69mmol), diiodomethane (1.08g, 4.03mmol) and cesium carbonate (1.31g, 4.03mmol) were dissolved in DMF (10 mL) and the reaction was reacted at 110 ℃ for 1.5 hours. After completion of the reaction, it was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/80% petroleum ether) to give 5-fluoro-2H-1,3-benzodioxolane-4-carboxylic acid methyl ester 199e (320 mg, yellow solid), yield: 54.11 percent.
MS m/z(ESI)：199.1(M+1).
The fifth step
Preparation of (5-fluoro-2H-1,3-benzodioxol-4-yl) methanol 199f
5-fluoro-2H-1,3-benzodioxolane-4-carboxylic acid methyl ester 199e (300mg, 1.514mmol) was dissolved in 5mL THF, followed by addition of 5mL LiAlH under an ice-water bath 4 Was added to the reaction solution, and the mixture solution was stirred at 25 ℃ for 2 hours. After completion of the reaction, it was extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 30% ethyl acetate/70% petroleum ether) to give (5-fluoro-2H-1,3-benzodioxolan-4-yl) methanol 199f (120 mg, white solid), yield: 41.93 percent.
MS m/z(ESI)：153.1(M-18).
The sixth step
Preparation of 4- (bromomethyl) -5-fluoro-2H-1,3-benzodioxane 199g
(5-fluoro-2H-1,3-benzodioxol-4-yl) methanol 199f (100mg, 0.5878mmol) and PBr 3 (190.93mg, 0.7053mmol) was dissolved in 5mL DCM, and the mixture solution was stirred at 25 ℃ for 1h. After completion of the reaction, it was extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 30% ethyl acetate/70% petroleum ether) to give 199g (100 mg, white solid) of 4- (bromomethyl) -5-fluoro-2H-1,3-benzodioxane, yield: 52.57 percent.
1H NMR(400MHz,cdcl3)δ6.68(dd,J＝8.5,4.3Hz,1H),6.53(dd,J＝10.2,8.6Hz,1H),6.06(s,2H),4.48(s,2H).
Seventh step
Preparation of 5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2H-1,3-benzodioxane 199H
4-fluoro-2-methoxy-5-nitrophenol (100mg, 0.5344mmol) was dissolved in 20mL DMF and NaH (25.65mg, 1.0688mmol) was added at 0 deg.C and stirred at that temperature for 10 minutes, then 4- (bromomethyl) -5-fluoro-2H-1,3-benzodioxane 199g (124.53mg, 0.5344mmol) was added and stirred for 10 minutes and reacted at 25 deg.C for 1 hour. After completion of the reaction, the reaction system was made acidic with 2N diluted hydrochloric acid, and extracted with ethyl acetate (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: 10% ethyl acetate/90% petroleum ether) to give 5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2H-1,3-benzodioxane 199H (50 mg, yellow solid), yield: 19.85 percent.
MS m/z(ESI)：340.2(M+1)。
Eighth step
Preparation of 2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxol-4-yl) methoxy ] -4-methoxyaniline 199i
5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2H-1,3-benzodioxane 199H (50mg, 0.1474 mmol) was dissolved in methanol (10 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (82.32mg, 1.474mmol) were added, and the reaction mixture was reacted at 80 ℃ for 1 hour. Then, it was filtered, and the filtrate was extracted with methylene chloride (3x20 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give 2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxol-4-yl) methoxy ] -4-methoxyaniline 199i (40 mg, yellow solid) in yield: 78.97%.
MS m/z(ESI)：310.0(M+1)。
The ninth step
Preparation of 4- [ ({ 2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxolen-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester 199j
2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxol-4-yl) methoxy ] -4-methoxyaniline 199i (40mg, 0.1293 mmol) and 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester (86.72mg, 0.2586mmol) were dissolved in tetrahydrofuran (10 mL). Then, triethylamine (39.25mg, 0.3879 mmol) was added to the reaction solution, and the reaction solution was reacted at 25 ℃ for 16 hours. The reaction solution was not subjected to any post-treatment to give a crude 4- [ ({ 2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid methyl ester 199j.
MS m/z(ESI)：551.1(M+1)。
The tenth step
Preparation of 3- (2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxolan-4-yl) methoxy ] -4-methoxyphenyl) -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
4- [ ({ 2-fluoro-5- [ (5-fluoro-2H-1,3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2,3-dicarboxylic acid dimethyl ester 199j (5 mL stock solution) was dissolved in methanol with lithium hydroxide (6.52mg, 0.2724mmol): water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 2 hours. After the reaction was completed, the reaction mixture was directly concentrated and purified by reverse column (water: acetonitrile = 20).
MS m/z(ESI)：505.1(M+1)。
HPLC:98.72％(214nm),96.20％(254nm).
1 H NMR(400MHz,dmso)δ7.19(d,J＝7.4Hz,1H),7.09(d,J＝11.6Hz,1H),6.94(dd,J＝8.6,4.4Hz,2H),6.71(dd,J＝10.4,8.6Hz,1H),6.09(d,J＝7.1Hz,2H),4.91(s,2H),3.80(s,3H).
Example 109
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002001
First step of
Preparation of 2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene
(6- (cyclopropylmethoxy) -2,3-difluorophenyl) methanol 201a (12g, 0.056 mol) was dissolved in dichloromethane (100 mL), triphenylphosphine (17.63g, 0.067 mol) and carbon tetrabromide (22.29g, 0.067 mol) were added thereto, and the reaction mixture was stirred at room temperature for 1 hour, after the completion of the reaction, the reaction mixture was concentrated and then separated and purified by a silica gel column (petroleum ether) to give 2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorophenyl 201b (9 g, white solid) in a yield of 58%.
1 H NMR(400MHz,CDCl3)δ7.04(dd,J＝19.0,9.2Hz,1H),6.54(ddd,J＝9.2,3.5,2.1Hz,1H),4.60(d,J＝1.7Hz,2H),3.87(d,J＝6.8Hz,2H),1.33–1.24(m,1H),0.68–0.62(m,2H),0.42–0.36(m,2H).
Second step of
Preparation of 1- (cyclopropylmethoxy) -3,4-difluoro-2- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzene
2- (bromomethyl) -1- (cyclopropylmethoxy) -3,4-difluorobenzene 201b (3g, 0.011mol) and 4-fluoro-2-methoxy-5-nitrophenol (2.02g, 0.011mol) were dissolved in acetonitrile (100 mL), followed by addition of potassium carbonate (4.48g, 0.032mol), potassium iodide (1.79g, 0.011mol), and the reaction mixture was stirred at 70 ℃ for 16 hours, after completion of the reaction, water was added to the reaction mixture, ethyl acetate was extracted, and the organic phase was dried and concentrated, and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 1- (cyclopropylmethoxy) -3,4-difluoro-2- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzene 201c (1 g, yellow solid) in 24% yield.
MS m/z(ESI)：384.0(M+1)。
The third step
Preparation of 5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline
1- (cyclopropylmethoxy) -3,4-difluoro-2- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzene 201c (1g, 2.61mmol) was dissolved in ethanol/ammonium chloride aqueous solution =4/1 (50 mL), iron powder (1.46g, 26.1mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated and then separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline 201d (850 mg, yellow oily liquid) in 92% yield.
MS m/z(ESI)：354.1(M+1)。
The fourth step
Preparation of 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline 201d (850mg, 2.41mmol) was dissolved in tetrahydrofuran (20 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (887mg, 2.65mmol) and triethylamine (730mg, 7.22mmol), the reaction solution was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction solution was concentrated, and after separation and purification by a silica gel column (petroleum ether/ethyl acetate = 3/1), 4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 201e (1.3 g, yellow solid) was obtained in 91% yield.
MS m/z(ESI)：595.1(M+1)。
The fifth step
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 201e (1.3g, 2.18mmol) was dissolved in tetrahydrofuran/water =1/1 (20 mL), lithium hydroxide (0.1lg, 6.54mmol) was added, the reaction solution was stirred at room temperature for 1 hour, tetrahydrofuran was removed after the reaction was completed, dilute hydrochloric acid was used to adjust the pH to acidic, ethyl acetate was extracted, and the extracted organic phase was separated and purified by a silica gel column (dichloromethane/methanol = 50/1) to give 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-900 mg, yield 75% yellow solid.
MS m/z(ESI)：549.1(M+1)。
HPLC:99.40％(214nm),100％(254nm)。
1 H NMR(400MHz,DMSO)δ14.52(s,1H),11.99(s,1H),7.48–7.36(m,2H),7.25(d,J＝7.4Hz,1H),7.14(d,J＝11.6Hz,1H),6.91–6.85(m,1H),5.01(s,2H),3.86(d,J＝6.9Hz,2H),3.82(s,3H),1.20–1.11(m,1H),0.53–0.46(m,2H),0.29–0.22(m,2H).
Example 110
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxa-1,2,3,4-tetrahydrothieno [2,3-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002021
First step of
Preparation of 2-aminothiophene-3,4-dicarboxylic acid ethyl ester
Ethyl 2-cyanoacetate 202a (10g, 88.4mmol), ethyl 2-oxopropionate (10.26g, 88.4mmol) and sulfur (2.83g, 88.6 mmol) were dissolved in N, N-dimethylformiate (50 mL), and triethylamine (9.84g, 97.2mmol) was added to the solution under ice-bath conditions. The reaction solution was stirred at 50 ℃ for 3 hours. After the reaction was complete, the reaction was quenched with water (20 mL), extracted with methyl tert-butyl ether (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on silica gel column (petroleum ether/ethyl acetate = 28%) to give 2-aminothiophene-3,4-dicarboxylic acid ethyl ester 202b (16 g, yellow solid), yield: 73 percent.
MS m/z(ESI)：244.0(M+1)。
Second step of
Preparation of 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3,4-dicarboxylic acid diethyl ester
Phenyl p-nitrochloroformate (4.96g, 24.6 mmol) and sodium hydrogencarbonate (2.07g, 24.6 mmol) were dissolved in acetonitrile (20 mL), and 2-aminothiophene-3,4-dicarboxylic acid diethyl ester 202b (3g, 12.3 mmol) was added to the solution under ice-bath conditions. The reaction solution was stirred at 25 ℃ for 6 hours. After the reaction, the reaction solution was filtered and concentrated to obtain a crude product 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3,4-dicarboxylic acid diethyl ester 202c, which was used in the next step without treatment.
MS m/z(ESI)：409.0(M+1)。
The third step
Preparation of 2- (3- (5- ((6-cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-3,4-dicarboxylic acid diethyl ester
The crude product obtained, diethyl 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3,4-dicarboxylate 202c (1.89g, 4.6 mmol), 5- ((6-cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline (1.5g, 4.2mmol) and triethylamine (850mg, 8.4mmol) were dissolved in tetrahydrofuran (20 mL). The reaction solution was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified on a silica gel column (petroleum ether/ethyl acetate = 23%) to give 2- (3- (5- ((6-cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-3,4-dicarboxylic acid diethyl ester 202d (2.7 g, yellow solid), yield: 85 percent.
MS m/z(ESI)：623.2(M+1)。
The fourth step
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxa-1,2,3,4-tetrahydrothieno [2,3-d ] pyrimidine-5-carboxylic acid
Diethyl 2- (3- (5- ((6-cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-3,4-dicarboxylate 202d (300mg, 0.48mmol) was dissolved in tetrahydrofuran (5 mL) and then sodium hydride (35mg, 1.44mmol) was added while cooling on ice. The reaction was stirred for 7 hours under ice bath. After completion of the reaction, the reaction solution was quenched with methanol (10 mL), washed with water (10 mL), extracted with ethyl acetate (3 × 10 mL), the combined organic layers were washed with brine (10 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA, mobile phase: ACN: H) 2 O =53%: 47%) to give 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxa-1,2,3,4-tetrahydrothieno [2,3-d]Pyrimidine-5-carboxylic acid Cpd-115 (14.78 mg, white solid), yield: 5.60 percent.
MS m/z(ESI)：549.0(M+1)。
HPLC:98.65％(214nm),99.21％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ14.16(s,1H),7.94(s,1H),7.43(dd,J＝19.6,9.6Hz,1H),7.24(d,J＝7.4Hz,1H),7.13(d,J＝11.6Hz,1H),6.94–6.86(m,1H),5.00(d,J＝9.8Hz,2H),3.86(d,J＝6.8Hz,2H),3.81(s,3H),1.19–1.10(m,1H),0.52–0.45(m,2H),0.28–0.22(m,2H).
Example 111
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [2,3-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340002031
First step of
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [2,3-d ] pyrimidine-2,4 (1H, 3H) -dione
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxa-1,2,3,4-tetrahydrothieno [2,3-d]Pyrimidine-5-carboxylic acid 203a (160mg, 0.29mmol) was dissolved in tetrahydrofuran (5 mL) and a solution of lithium aluminum hydride in tetrahydrofuran (0.3mL, 0.29mmol) was added to the solution under ice-bath conditions. The reaction solution was stirred at 0 ℃ for 1 hour. After the reaction was complete, the reaction was quenched with methanol (20 mL), quenched with water (20 mL), extracted with ethyl acetate (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by preparative purification (FA, mobile phase: ACN: H) 2 O =50%: 50%) to give 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [2,3-d]Pyrimidine-2,4 (1h, 3h) -dione Cpd-116 (6.38 mg, white solid), yield: 4.01 percent.
MS m/z(ESI)：535.0(M+1)。
HPLC:96.63％(214nm),98.25％(254nm).
1 H NMR(400MHz,DMSO-d 6 )δ7.43(dd,J＝19.6,9.6Hz,1H),7.14(d,J＝7.4Hz,1H),7.05(d,J＝11.2Hz,1H),6.91–6.85(m,1H),6.76(s,1H),5.00(s,2H),4.59(s,2H),3.86(d,J＝6.8Hz,2H),3.79(s,3H),1.18–1.08(m,1H),0.53–0.43(m,2H),0.29–0.19(m,2H).
Example 112
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
Figure BDA0003608368340002041
First step of
Preparation of 3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- (hydroxymethyl) thieno [3,4-d ] pyrimidine-2,4 (1H, 3H) -dione
3- (5- ((6- (cyclopropylmethoxy) -2,3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-114 (300mg, 0.55mmol) was dissolved in tetrahydrofuran (5 mL), 2M lithium aluminum hydrogen in tetrahydrofuran solution (0.55mL, 1.1mmol) was slowly added dropwise at 0 deg.C, the reaction was stirred at room temperature for 2 hours, ammonium chloride aqueous solution was quenched, ethyl acetate was extracted, and the organic phase was dried to afford Cpd-117 (25 mg, yellow solid) in 8% yield.
MS m/z(ESI)：535.0(M+1)。
HPLC:92.66％(214nm),86.70％(254nm)。
1 H NMR(400MHz,DMSO)δ11.41(s,1H),7.49(dd,J＝19.7,9.5Hz,1H),7.25(d,J＝7.4Hz,1H),7.13(d,J＝11.5Hz,1H),6.97–6.92(m,1H),6.78(s,1H),6.04(s,1H),5.13–5.01(m,4H),3.92(d,J＝6.9Hz,2H),3.85(s,3H),1.26–1.16(m,1H),0.57–0.51(m,2H),0.35–0.29(m,2H).
Example 113
3- [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002042
First step of
Preparation of 4-fluoro-5-nitrobenzene-1,2-diol 205b
To 4-fluoro-2-methoxy-5-nitrophenol 205a (2g, 0.0107 mol) were added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120 ℃ for 12h. After completion of the reaction, it was cooled to room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified by column chromatography (mobile phase: 20% ethyl acetate/80% petroleum ether) to give 4-fluoro-5-nitrobenzene-1,2-diol 205b (1.1 g, yellow solid) in yield: and 59.81 percent.
MS m/z(ESI)：174.1(M+1).
Second step of
Preparation of 2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 205c
4-fluoro-5-nitrobenzene-1,2-diol 205b (100mg, 0.5777mmol) was dissolved in 20mL DMF, naH (27.73mg, 1.1554mmol) was added at 0 ℃ and stirred at that temperature for 10 minutes, then 2-bromo-3,4-difluoro-1-methoxybenzene (128.84mg, 0.5777mmol) was added and stirred for 10 minutes and reacted at 25 ℃ for 1 hour. After completion of the reaction, the reaction system was made acidic with 2N diluted hydrochloric acid, and extracted with ethyl acetate (2X 20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: 10% ethyl acetate/90% petroleum ether) to give 2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 205c (100 mg, yellow solid), yield: 43.93 percent.
MS m/z(ESI)：330.1(M+1)。
The third step
Preparation of 1- (cyclopropylmethoxy) -2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluoro-4-nitrobenzene 205d
2- (2,3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 205c (100mg, 0.3035 mmol), (bromomethyl) cyclopropane (40.95mg, 0.3035 mmol), potassium carbonate (5.05mg, 0.003mmol) were dissolved in acetonitrile (10 mL) and stirred at 80 ℃ for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with EA (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: 10% ethyl acetate/90% petroleum ether) to give 1- (cyclopropylmethoxy) -2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluoro-4-nitrobenzene 205d (100 mg, yellow solid) in yield: 77.43 percent.
MS m/z(ESI)：384.2(M+1)。
The fourth step
Preparation of 4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoroaniline 205e
1- (Cyclopropylmethoxy) -2- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -5-fluoro-4-nitrobenzene 205d (100mg, 0.2609mmol) was dissolved in methanol (10 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (145.71mg, 2.609mmol) were added, and the reaction solution was reacted at 80 ℃ for 1 hour. Then filtered and the filtrate extracted with dichloromethane (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoroaniline 205e (100 mg, yellow solid), yield: 97.62 percent.
MS m/z(ESI)：354.1(M+1)。
The fifth step
Preparation of 4- ({ [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 205f
4- (Cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoroaniline 205e (100mg, 0.283mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2,3-dicarboxylate (189.8mg, 0.566mmol) were dissolved in tetrahydrofuran (10 mL). Then, triethylamine (85.91mg, 0.849 mmol) was added thereto, and the reaction mixture was reacted at 25 ℃ for 16 hours. The reaction solution was not subjected to any post-treatment to give a crude 4- ({ [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 205f.
MS m/z(ESI)：595.1(M+1)。
The sixth step
Preparation of 3- [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dissolving 4- ({ [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2,3-dicarboxylic acid dimethyl ester 205f (5 mL stock solution) and lithium hydroxide (12.08mg, 0.5046mmol) in methanol: water =3:1 (4 mL). The reaction mixture was reacted at 25 ℃ for 2 hours. After the reaction was completed, the reaction mixture was directly concentrated and purified by reverse column (water: acetonitrile = 20) to give 3- [4- (cyclopropylmethoxy) -5- [ (2,3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] -2,4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-118 (1.17 mg, yellow solid) in 1.25% yield.
MS m/z(ESI)：549.1(M+1)。
1 H NMR(400MHz,dmso)δ8.29(s,1H),7.39(dd,J＝19.6,9.56Hz,1H),7.04(d,J＝7.56Hz,1H),6.95(d,J＝11.6Hz,1H),6.88–6.79(m,1H),4.91(s,2H),3.77(d,J＝7.0Hz,2H),3.73(s,3H),0.77(t,J＝6.8Hz,1H),0.50–0.38(m,2H),0.21(q,J＝5.0Hz,2H).
Example 114
3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002061
First step of
Preparation of 2- ((2-butoxy-4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene
2- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-fluoro-4-nitrophenol 206a (150mg, 0.4556mmol) was dissolved in acetonitrile (10 mL), and 1-bromobutane 206b (74.91mg, 0.54672mmol), potassium carbonate (125.94mg, 0.9112mmol), potassium iodide (7.56mg, 0.04556mmol) were added. The reaction was carried out at 90 ℃ for 2 hours, and after completion of the reaction, the temperature was returned to room temperature. Acetonitrile was spin-dried and the residue was passed through a silica gel column (petroleum ether/ethyl acetate = 10/1) to give 2- ((2-butoxy-4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene 206c (150 mg, yellow solid). Yield: 83.74%.
MS m/z(ESI)：386.3(M+1)。
Second step of
Preparation of 4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline
2- ((2-butoxy-4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene 206c (150mg, 0.3893mmol) was dissolved in a mixture of ethanol/water =1/1 (10 mL), and iron powder (43.48mg, 0.7786mmol), ammonium chloride (62.47mg, 1.1679mmol) was added. The reaction was carried out at 80 ℃ for 1 hour. After the reaction was completed, the temperature was returned to room temperature. And (5) filtering. The ethanol was spun off and ethyl acetate (3 × 10 mL) was added. And (4) extracting. The organic phase was collected, dried, filtered and concentrated. The concentrate was subjected to a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline 206d (100 mg, yellow solid). Yield: 70.85%.
MS m/z(ESI)：356.3(M+1)。
The third step
Preparation of dimethyl 4- (3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylate
4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline 206d (100mg, 0.2814 mmol) was dissolved in tetrahydrofuran (5 mL), and 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 206e (188.72mg, 0.5628mmol), triethylamine (56.95mg, 0.5628mmol) were added. The reaction was carried out at room temperature for 16 hours. After the reaction is finished. The solvent was dried by spinning and the concentrate was passed through a silica gel column (petroleum ether/ethyl acetate = 2/1) to give 4- (3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 206f (50 mg, yellow solid). Yield: 29.18%.
MS m/z(ESI)：597.5(M+1)。
The fourth step
Preparation of 3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 206f (50mg, 0.0838mmol) was dissolved in a mixed solution of tetrahydrofuran/water =1/1 (5 mL), and lithium hydroxide (4.01mg, 0.1676mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the solvent was spin dried. The raffinate was passed through a preparative column (acetonitrile/water = 62/38) to give 3- (4-butoxy-5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-119 (30.03 mg, white solid). Yield: 63.84 percent.
MS m/z(ESI)：551.5(M+1)。
HPLC:97.32％(214nm),95.08％(254nm).
1 H NMR(400MHz,dmso)δ14.55(s,1H),11.96(s,1H),7.46(dd,J＝19.7,9.3Hz,1H),7.34(s,1H),7.21(d,J＝7.5Hz,1H),7.13(d,J＝11.7Hz,1H),6.89(d,J＝9.3Hz,1H),4.99(s,2H),4.02(t,J＝6.5Hz,2H),3.79(s,3H),1.74–1.63(m,2H),1.41(dd,J＝14.9,7.5Hz,2H),0.91(t,J＝7.4Hz,3H).
Example 115
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002071
First step of
Preparation of 1,2-difluoro-3- ((4-fluoro-2- (2-methoxyethoxy) -5-nitrophenoxy) methyl) -4-methoxybenzene
2- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-fluoro-4-nitrophenol 207a (150mg, 0.456mmol) was dissolved in acetonitrile (5 mL), followed by the addition of 1-bromo-2-methoxyethane (127mg, 0.912mmol), potassium iodide (76mg, 0.456mmol) and potassium carbonate (126mg, 0.912mmol) at 80 ℃, the reaction was quenched with stirring for 2 hours, after the reaction was complete, the reaction was filtered, and the filtrate was spin-dried and purified by silica gel column (petroleum ether/ethyl acetate = 5/1) to give 1,2-difluoro-3- ((4-fluoro-2- (2-methoxyethoxy) -5-nitrophenoxy) methyl) -4-methoxybenzene 207b (100 mg, yellow solid) in 56.6% yield.
MS m/z(ESI)：410.1(M+Na)。
Second step of
Preparation of 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) aniline
1,2-difluoro-3- ((4-fluoro-2- (2-methoxyethoxy) -5-nitrophenoxy) methyl) -4-methoxybenzene 207b (100mg, 0.258mmol) was dissolved in ethanol/water =4/1 (5 mL), followed by addition of iron powder (144mg, 2.582mmol), the reaction was stirred at 80 ℃ for 1 hour, and after completion of the reaction, the reaction was spin-dried and purified by silica gel column (petroleum ether/ethyl acetate = 5/1) to give 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) aniline 207c (70 mg, yellow oily liquid) in 75.9% yield.
MS m/z(ESI)：357.9(M+1)。
The third step
Preparation of 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) aniline 207c (70mg, 0.193mmol) was dissolved in tetrahydrofuran (3 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (71mg, 0.21201 mmol) and triethylamine (58mg, 0.579mmol), the reaction was stirred at room temperature for 16 hours, and at the end of the reaction, the reaction was concentrated and purified by silica gel column (petroleum ether/ethyl acetate = 2/1) separation to give 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 207d (85 mg, yellow solid) in 73.6% yield.
MS m/z(ESI)：599.1(M+1)。
The fourth step
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid
The isolated and purified 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 207d (85mg, 0.142mmol) was dissolved in tetrahydrofuran/water =1/1 (2 mL), lithium hydroxide (10mg, 0.426mmol) was then added, the reaction solution was stirred at room temperature for 1 hour, and after the reaction was completed, the reaction solution was concentrated to produce 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid Cpd-120 (33 mg, yellow solid) in 42.1% yield.
MS m/z(ESI)：553.1(M+1)。
HPLC:99.36％(214nm),100％(254nm)。
1 H NMR(400MHz,dmso)δ11.95(s,1H),7.46(dd,J＝19.7,9.4Hz,1H),7.31(s,1H),7.23(d,J＝7.5Hz,1H),7.16(d,J＝11.6Hz,1H),6.90(ddd,J＝9.3,3.5,1.8Hz,1H),4.99(s,2H),4.21–4.13(m,2H),3.80(s,3H),3.69–3.62(m,2H),3.29(s,3H).
Example 116
3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002081
First step of
Preparation of 3- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-methoxy-5-nitropyridine
3-bromo-2-methoxy-5-nitropyridine 208a (500mg, 2.1457mmol) was dissolved in 1,4-dioxane (10 mL) and (2,3-difluoro-6-methoxyphenyl) methanol 208b (448.39mg, 2.57484mmol), tris (dibenzylideneacetone) dipalladium (123.38mg, 0.21457mmol), 2- (dicyclohexylphosphine) -3,6-dimethoxy-2 '-4' -6 '-tri-I-propyl-11' -biphenyl (172.76mg, 0.321855mmol), cesium carbonate (1398.22mg, 4.2914mmol) were added. The reaction was carried out at 90 ℃ for 16 hours under nitrogen. After the reaction is finished. Water and ethyl acetate were added. Extracting, drying and concentrating. The concentrate was passed through a silica gel column (petroleum ether/ethyl acetate = 10/1) to give 3- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-methoxy-5-nitropyridine 208c (150 mg, yellow solid). Yield: 21.00 percent.
MS m/z(ESI)：327.2(M+1)
Second step of
Preparation of 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-amine
3- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-methoxy-5-nitropyridine 208c (150mg, 0.4598mmol) was dissolved in a mixed solution of ethanol/water =1/1 (10 mL), and iron powder (51.36mg, 0.9196 mmol), ammonium chloride (73.78mg, 1.3794mmol) were added. Reacting at 80 deg.C for 1 hr, recovering to room temperature after reaction, removing ethanol, extracting with ethyl acetate, collecting organic phase, drying, filtering, and concentrating. The concentrate was passed through a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-amine 208d (90 mg, yellow liquid). Yield: 62.77%.
MS m/z(ESI)：297.2(M+1)
The third step
Preparation of dimethyl 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) ureido) thiophene-2,3-dicarboxylate
5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-amine 208d (90mg, 0.3038mmol) was dissolved in tetrahydrofuran (5 mL), and 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 208e (122.25mg, 0.36456mmol), triethylamine (61.48mg, 0.6076mmol) were added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried by spinning, and the residue was subjected to a silica gel column (petroleum ether/ethyl acetate = 1/1) to give 4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 208f (10 mg, yellow liquid). Yield: 5.99 percent.
MS m/z(ESI)：538.4(M+1)。
The fourth step
Preparation of 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 208f (10mg, 0.0186mmol) was dissolved in a tetrahydrofuran/water =1/1 (3 mL) mixed solution, and lithium hydroxide (0.89mg, 0.0372mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the solvent was spun off and the residue was passed through preparative column (acetonitrile/water = 49/51) to give 3- (5- ((2,3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-121 (3.47 mg, yellow solid). Yield: 37.10 percent.
MS m/z(ESI)：492.4(M+1)
HPLC:99.22％(214nm),92.32％(254nm).
1 H NMR(400MHz,DMSO)δ8.29(s,1H),7.63(s,1H),7.56–7.45(m,2H),6.93(d,J＝10.3Hz,1H),6.54(s,1H),5.00(s,2H),3.87(s,3H),3.82(s,3H).
Example 117
3- (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002101
First step of
Preparation of tert-butyl (4-chloro-5-methoxypyridin-2-yl) carbamate
4-chloro-5-methoxypyridin-2-amine 209a (400mg, 2.5223mmol) was dissolved in methylene chloride (5 mL), di-tert-butyl dicarbonate (1100.98mg, 5.0446mmol) and triethylamine (765.69mg, 7.5669mmol) were added to the solution, and the mixture was reacted at room temperature for 1 hour, followed by addition of water (10 mL) after completion of the reaction, extraction, drying and concentration. The concentrate was then subjected to a silica gel column (petroleum ether/ethyl acetate = 10/1) to give tert-butyl (4-chloro-5-methoxypyridin-2-yl) carbamate 209b (270 mg, white solid). Yield: 41.38 percent.
MS m/z(ESI)：259.1(M+1)。
Second step of
Preparation of tert-butyl (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) carbamate
(4-chloro-5-methoxypyridin-2-yl) carbamic acid tert-butyl ester 209b (270mg, 1.0437mmol) was dissolved in 1,4-dioxane (10 mL), and (2,3-difluoro-6-methoxyphenyl) methanol 209c (218.11mg, 1.25244mmol), tris (dibenzylideneacetone) dipalladium (95.57mg, 0.10437mmol), 2- (dicyclohexylphosphine) -3,6-dimethoxy-2 '-4' -6 '-tri-I-propyl-11' -biphenyl (84.03mg, 0.156555mmol), cesium carbonate (680.12mg, 2.0874mmol) were added. The reaction was carried out at 90 ℃ for 16 hours under nitrogen. After the reaction is finished. Water and ethyl acetate were added. Extracting, drying and concentrating. The concentrate was subjected to silica gel column (petroleum ether/ethyl acetate = 10/1) to give tert-butyl (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) carbamate 209d (100 mg, white solid). Yield: 24.17 percent.
MS m/z(ESI)：397.1(M+1)。
The third step
Preparation of 4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-amine
Tert-butyl (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) carbamate 209d (100mg, 0.2523mmol) was added to a mixed solution of dichloromethane/trifluoroacetic acid =4/1 (5 mL) at 0 ℃, followed by stirring at 0 ℃ for 1 hour. After the reaction was complete, the solvent was evaporated and the residue was passed through a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-amine 209e (60 mg, yellow liquid). Yield: 80.26 percent.
MS m/z(ESI)：297.0(M+1)。
The fourth step
Preparation of dimethyl 5- (3- (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) ureido) thiophene-2,3-dicarboxylate
4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-amine 209e (60mg, 0.2025mmol) was dissolved in tetrahydrofuran (5 mL), and 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 209f (135.81mg, 0.405mmol), triethylamine (40.98mg, 0.405mmol) were added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried by spinning, and the residue was subjected to a silica gel column (petroleum ether/ethyl acetate = 1/1) to obtain 209g (62 mg, yellow liquid) of 5- (3- (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester. Yield: 56.99%.
MS m/z(ESI)：538.0(M+1)。
The fifth step
Preparation of 3- (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
209g (62mg, 0.1154 mmol) of dimethyl 5- (3- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) ureido) thiophene-2,3-dicarboxylate was dissolved in a mixed solution of tetrahydrofuran/water =1/1 (5 mL). Lithium hydroxide (5.53mg, 0.2308mmol) was added thereto, and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the solvent was spun off and the raffinate was passed through preparative column (acetonitrile/water = 46/54) to give 3- (4- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-122 (1.35 mg, white solid). Yield: 2.34 percent.
MS m/z(ESI)：492.4(M+1)。
HPLC:100.00％(214nm),97.69％(254nm).
1 H NMR(400MHz,DMSO)δ8.39(s,1H),8.12(s,1H),7.52(q,J＝9.6Hz,1H),6.95(d,J＝8.8Hz,1H),6.53(s,1H),5.07(s,2H),3.17(s,3H).
Example 118
3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002111
First step of
Preparation of 2-methoxy-6- (trifluoromethyl) benzaldehyde
A solution of 2-fluoro-6- (trifluoromethyl) benzaldehyde 210a (900mg, 4.66mmol) and potassium hydroxide (915mg, 16.31mmol) in methanol (15 mL) was stirred at 25 ℃ for 1 hour. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 2-methoxy-6- (trifluoromethyl) benzaldehyde 210b (890 mg, white solid), yield: 83.78%.
MS m/z(ESI):205.1(M+1)。
Second step of
Preparation of (2-methoxy-6- (trifluoromethyl) phenyl) methanol
A solution of 2-methoxy-6- (trifluoromethyl) benzaldehyde 210b (890 mg, 4.33mmol) and sodium borohydride (180mg, 4.77mmol) in methanol (15 mL) was stirred at 25 ℃ for 1 hour. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 7/3) to give (2-methoxy-6- (trifluoromethyl) phenyl) methanol 210c (1.7 g, white solid) in yield: 94.58%.
MS m/z(ESI):189.1(M+1-18)。
The third step
Preparation of 2- (bromomethyl) -1-methoxy-3- (trifluoromethyl) benzene
(2-methoxy-6- (trifluoromethyl) phenyl) methanol 210c (1.7g, 0.01mol) was dissolved in dichloromethane (20 mL). Phosphorus tribromide (2.73g, 0.01mol) was added dropwise to the reaction at 0 ℃. The reaction was stirred at 0 ℃ for 2 hours. After the reaction was complete, the reaction was quenched by addition of saturated aqueous sodium bicarbonate and extracted with dichloromethane (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 100/1) to give 2- (bromomethyl) -1-methoxy-3- (trifluoromethyl) benzene 210d (770 mg, clear oil), yield: 30.05 percent.
1 H NMR(400MHz,CDCl 3 )δ7.42-7.38(m,1H),7.26(d,J＝8.4Hz,1H),7.10(d,J＝8.4Hz,1H),4.68(s,2H),3.97(s,3H)..
The fourth step
Preparation of 1-fluoro-5-methoxy-4- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene
A solution of 2- (bromomethyl) -1-methoxy-3- (trifluoromethyl) benzene 210d (770mg, 2.85mmol), 4-fluoro-2-methoxy-5-nitrophenol 210e (693mg, 3.70mmol), potassium carbonate (788mg, 5.70mmol), and potassium iodide (236mg, 1.42mmol) in acetonitrile (10 mL) was stirred at 80 ℃ for 16 hours. After the reaction was complete, quench with water and extract with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 1-fluoro-5-methoxy-4- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene 210f (680 mg, white solid) in yield: 57.04%.
MS m/z(ESI):376.9(M+1)。
The fifth step
Preparation of 2-fluoro-4-methoxy-5- (2-methoxy-6- (trifluoromethyl) benzyl) oxy) aniline
1-fluoro-5-methoxy-4- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene 210f (680mg, 1.80mmol), iron powder (302mg, 5.42mmol) and ammonium chloride (290mg, 5.42mmol) in ethanol: water =1:1 (20 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 210g (490 mg, white solid) of 2-fluoro-4-methoxy-5- (2-methoxy-6- (trifluoromethyl) benzyl) oxy) aniline, yield: 70.47%.
MS m/z(ESI):346.0(M+1)。
The sixth step
Preparation of 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
A solution of 210g (470mg, 1.35mmol) of 2-fluoro-4-methoxy-5- (2-methoxy-6- (trifluoromethyl) benzyl) oxy) aniline, 4- (phenoxycarbonyl) amino) thiophene-2,3 dimethyl ester, 211h (546mg, 1.62mmol), and triethylamine (274mg, 2.71mmol) in tetrahydrofuran (20 mL) was stirred at 25 ℃ for 16 h. After the reaction was completed, water was added to quench, extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 211i (330 mg, red oil), yield: 37.25 percent.
MS m/z(ESI)：587.1(M+1)。
Seventh step
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylate 211i (310mg, 0.52mmol) and lithium hydroxide (37.9 mg, 1.58mmol) in tetrahydrofuran: water =1:1 (14 mL) was stirred at 25 ℃ for 1 hour. After the reaction, the reaction mixture was concentrated. The concentrate was lyophilized by preparative purification to give 3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-123 (60 mg, yellow solid), yield: 20.58 percent.
MS m/z(ESI)：541.1(M+1)。
HPLC:100％(214nm),98.52％(254nm).
1 H NMR(400MHz,DMSO-d6)δ7.66-7.62(m,1H),7.45(d,J＝8.4Hz,1H),7.38(d,J＝7.9Hz,1H),7.26(d,J＝7.3Hz,1H),7.18(s,1H),7.09(d,J＝11.5Hz,1H),4.99(s,2H),3.87(s,3H),3.78(s,3H).
Example 119
3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002131
First step of
Preparation of (2-methoxy-5- (trifluoromethyl) phenyl) methanol
A solution of 2-methoxy-5- (trifluoromethyl) benzaldehyde 211a (400mg, 1.94mmol) and sodium borohydride (81.14mg, 2.14mmol) in methanol (6 mL) was stirred at 25 ℃ for 16 h. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 17/3) to give (2-methoxy-5- (trifluoromethyl) phenyl) methanol 211b (380 mg, clear oil) in yield: 84.67%.
MS m/z(ESI):189.1(M+1-18)。
Second step of
Preparation of 2- (bromomethyl) -1-methoxy-4- (trifluoromethyl) benzene
(2-methoxy-5- (trifluoromethyl) phenyl) methanol 211b (580 mg, 2.79mmol) was dissolved in dichloromethane (10 mL). PBr was washed at 0 deg.C 3 (909mg, 3.35mmol) was added dropwise to the reaction. The reaction was stirred at 0 ℃ for 2 hours. After the reaction was complete, it was quenched by addition of saturated aqueous sodium bicarbonate and extracted with dichloromethane (3 × 10 mL). The organic phase is dried over anhydrous sodium sulfate and filtered Filtration, concentration of the filtrate and purification by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 100/1) gave 2- (bromomethyl) -1-methoxy-4- (trifluoromethyl) benzene 211c (320 mg, clear oil), yield: 38.09%.
1 H NMR(400MHz,CDCl 3 )δ7.60–7.54(m,2H),6.95(d,J＝8.6Hz,1H),4.54(s,2H),3.95(s,3H).
The third step
Preparation of 1-fluoro-5-methoxy-4- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene
A solution of 2- (bromomethyl) -1-methoxy-4- (trifluoromethyl) benzene 211c (100mg, 0.37mmol), 4-fluoro-2-methoxy-5-nitrophenol 211d (90mg, 0.48mmol), potassium carbonate (102mg, 0.74mmol) and potassium iodide (30.74mg, 0.18mmol) in acetonitrile (5 mL) was stirred at 80 ℃ for 24 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 1-fluoro-5-methoxy-4- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene 211e (120 mg, clear oil), yield: 77.50%.
MS m/z(ESI):376.0(M+1)。
The fourth step
Preparation of 2-fluoro-4-methoxy-5- (2-methoxy-5- (trifluoromethyl) benzyl) oxy) aniline
1-fluoro-5-methoxy-4- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) -2-nitrobenzene 211e (130mg, 0.34mmol), iron powder (57mg, 1.03mmol) and ammonium chloride (55mg, 1.03mmol) in ethanol: water =1:1 (10 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to give 2-fluoro-4-methoxy-5- (2-methoxy-5- (trifluoromethyl) benzyl) oxy) aniline 211f (85 mg, yellow oil), yield: 63.94 percent.
MS m/z(ESI):326.1(M+1-20)。
The fifth step
Preparation of 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
A solution of 2-fluoro-4-methoxy-5- (2-methoxy-5- (trifluoromethyl) benzyl) oxy) aniline 211f (80mg, 0.23mmol), 4- (phenoxycarbonyl) amino) thiophene-2,3 dimethyl dicarboxylate 211g (92mg, 0.27mmol) and triethylamine (46mg, 0.46mmol) in tetrahydrofuran (5 mL) was stirred at 25 ℃ for 1 hour. After the reaction was complete, quench with water and extract with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 211h (100 mg, clear oil), yield: 66.32 percent.
MS m/z(ESI):587.1(M+1)。
The sixth step
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2,3-dicarboxylate 211h (170mg, 0.28mmol) and lithium hydroxide (20.79mg, 0.86mmol) in tetrahydrofuran: water =1:1 (4 mL) was stirred at 25 ℃ for 1 hour. After the reaction was completed, saturated aqueous ammonium chloride was added to quench, the pH was adjusted to 7, followed by extraction with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and lyophilized by preparative purification to give 3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-124 (60.81 mg, white solid), yield: 38.04%.
MS m/z(ESI)：541.0(M+1)。
HPLC:98.98％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d6)δ14.57(s,1H),12.00(s,1H),7.81(s,1H),7.73(d,J＝8.6Hz,1H),7.39(s,1H),7.27(dd,J＝22.6,7.9Hz,2H),7.15(d,J＝11.5Hz,1H),5.01(s,2H),3.87(d,J＝15.1Hz,6H).
Example 120
3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002151
First step of
Preparation of 2- ((2- (cyclopentyloxy) -4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene
A solution of 2- ((2,3-difluoro-6-methoxybenzyl) oxy) -5-fluoro-4-nitrophenol 212a (300mg, 0.91mmol), bromocyclopentane (407 mg, 2.73mmol), potassium carbonate (251mg, 1.82mmol) and potassium iodide (75mg, 0.45mmol) in acetonitrile (5 mL) was stirred at 80 ℃ for 1 h. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to give 2- ((2- (cyclopentyloxy) -4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene 212b (110 mg, yellow solid), yield: 27.35 percent.
MS m/z(ESI):398.1(M+1)。
Second step of
Preparation of 4- (cyclopentyloxy) -5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline
2- ((2- (cyclopentyloxy) -4-fluoro-5-nitrophenoxy) methyl) -3,4-difluoro-1-methoxybenzene 212b (120mg, 0.30mmol)), iron powder (50mg, 0.90mmol), and ammonium chloride (48mg, 0.90mmol) were mixed in ethanol: water =1:1 (12 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, quench with water and extract with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 4- (cyclopentyloxy) -5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline 212c (110 mg, white solid) in yield: 89.24 percent.
MS m/z(ESI):368.0(M+1)。
The third step
Preparation of dimethyl 4- (3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylate
A solution of 4- (cyclopentyloxy) -5- ((2,3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline 212c (50mg, 0.13mmol), 4- (phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 212d (54mg, 0.16mmol) and triethylamine (27mg, 0.27mmol) in tetrahydrofuran (4 mL) was stirred at 25 ℃ for 16 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: methanol/dichloromethane = 10/1) to give 4- (3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 212e (47 mg, brown oil), yield: 51.07%.
MS m/z(ESI):609.2(M+1)。
The fourth step
Preparation of 3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2,3-dicarboxylate 212e (37mg, 0.06mmol) and lithium hydroxide (4mg, 0.18mmol) in tetrahydrofuran: water =1:1 (4 mL) was stirred at 25 ℃ for 1 hour. After the reaction was completed, the reaction solution was concentrated and lyophilized by preparative purification to give 3- (4- (cyclopentyloxy) -5- (2,3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-125 (2.01 mg, yellow solid), yield: 4.77 percent.
MS m/z(ESI)：563.0(M+1)。
HPLC:92.16％(214nm),81.86％(254nm).
1 H NMR(400MHz,DMSO-d6)δ8.30(s,1H),7.47-7.40(m,1H),7.10(d,J＝7.6Hz,1H),7.03(d,J＝11.5Hz,1H),6.88(d,J＝8.9Hz,1H),6.62(s,1H),4.96(s,2H),4.82(s,1H),3.78(s,3H),2.02-1.96(m,1H),1.87(d,J＝5.4Hz,2H),1.67(s,4H),1.55(s,2H).
Example 121
3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002161
First step of
Preparation of 2,3-dihydro-1,4-benzodioxane-5-methyl alcohol
Lithium aluminum hydride (15 mL) was added dropwise to a solution of 2,3-dihydrobenzo [ b ] [1,4] dioxin-5-carboxylic acid 213a (1g, 0.01mol) in tetrahydrofuran (12 mL) at 0 ℃. The reaction mixture was stirred at 0 ℃ for 10 minutes and at 25 ℃ for a further 16 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 2,3-dihydro-1,4-benzodioxan-5-methyl alcohol 213b (970 mg, white solid), yield: 94.64 percent.
MS m/z(ESI):149.2(M+1-18)。
Second step of
Preparation of 5-bromomethyl-2,3-dihydro-1,4-benzodioxine
Phosphorus tribromide (1.7g, 6.28mmol) was dropped into a solution of 2,3-dihydro-1,4-benzodioxan-5-methyl alcohol 213b (870mg, 5.23mmol) in dichloromethane (10 mL) and stirred at 0 ℃ for 1 hour. After the reaction was complete, it was quenched by addition of saturated sodium bicarbonate solution and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 5-bromomethyl-2,3-dihydro-1,4-benzodioxine 213c (1 g, yellow solid) in yield: 75.04%.
1 H NMR(400MHz,CDCl 3 )δ6.91-6.89(m,1H),6.85–6.77(m,2H),4.52(s,2H),4.36-4.34(m,2H),4.29-4.27(m,2H).
The third step
Preparation of 5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxine
A solution of 5-bromomethyl-2,3-dihydro-1,4-benzodioxine 213c (900mg, 3.92mol), 4-fluoro-2-methoxy-5-nitrophenol 213d (955mg, 5.10mmol), potassium carbonate (1.08g, 7.85mmol) and potassium iodide (326mg, 1.96mmol) in acetonitrile (10 mL) was stirred at 80 ℃ for 16 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin 213e (770 mg, yellow solid), yield: 62.17 percent.
MS m/z(ESI):(M+1+23)。
The fourth step
Preparation of 5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline
5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin 213e (810mg, 2.41mmol), iron powder (404mg, 7.24mmol), and ammonium chloride (387mg, 7.24mmol) in ethanol: water =1:1 (12 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, quench with water and extract with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 213f (670 mg, yellow solid), yield: 81.75 percent.
MS m/z(ESI):306.1(M+1)。
The fifth step
Preparation of dimethyl 4- (3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
A solution of 5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 213f (570mg, 1.86mmol), 4- (phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 213g (751mg, 2.24mmol) and triethylamine (377mg, 3.73mmol) in tetrahydrofuran (15 mL) was stirred at 25 ℃ for 16 hours. After the reaction was completed, water was added to quench, and extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to give 4- (3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 213h (600 mg, brown oil), yield: 51.42 percent.
MS m/z(ESI):547.1(M+1)。
The sixth step
Preparation of 3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 213h (500mg, 0.91mmol), and lithium hydroxide (65mg, 2.74mmol) in tetrahydrofuran: water =1:1 (14 mL) was stirred at 25 ℃ for 1 hour. After the reaction, the reaction solution was concentrated. The concentrate was lyophilized by preparative purification to give 3- (5- ((2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-126 (355.86 mg, yellow solid), yield: 76.81 percent.
MS m/z(ESI)：501.0(M+1)。
HPLC:99.51％(214nm),98.82％(254nm).
1 H NMR(400MHz,DMSO-d6)δ14.52(s,1H),11.96(s,1H),7.38(s,1H),7.25(d,J＝7.4Hz,1H),7.12(d,J＝11.6Hz,1H),7.01-6.98(m,1H),6.87-6.84(m,2H),4.91(s,2H),4.25(d,J＝4.1Hz,4H),3.84(s,3H).
Example 122
3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002181
First step of
Preparation of 2- (2-bromo-4-fluorophenoxy) ethan-1-ol
After 2-bromo-4-fluorophenol 1 (5g, 0.0262mol), 2-bromoethanol 2 (3.93g, 0.0314mol) were dissolved in acetonitrile (50 mL), followed by addition of potassium carbonate (7.24g, 0.0524mol) and potassium iodide (0.43g, 0.0026mol), the reaction solution was stirred at 80 ℃ for 16 hours, after completion of the reaction, the reaction solution was suction-filtered, the filtrate was concentrated, and the filtrate was purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 2- (2-bromo-4-fluorophenoxy) ethan-1-ol 3 (5 g, white solid) in yield: 77.10%.
MS m/z(ESI):236.1(M+1)。
Second step of
Preparation of 6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine
2- (2-bromo-4-fluorophenoxy) ethan-1-ol 3 (1.2g, 0.0051mmol), palladium acetate (0.02mg, 0.0001mmol), t BuDavePhos (0.04g, 0.00012mmol), cesium carbonate (2.49g, 0.0077mmol) were added to toluene (20 mL) and reacted at 80 ℃ for 1 hour with a microwave, after completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1) to give 6-fluoro-2,3-dihydrobenzene [ b ] and ][1,4]Dioxin 4 (0.5 g, white solid), yield: 62.75 percent.
MS m/z(ESI):155.1(M+1)。
The third step
Preparation of 6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine-5-carbaldehyde
6-fluoro-2,3-dihydrobenzene [ b ] [1,4] dioxin 4 (200mg, 1.2975mmol) was dissolved in tetrahydrofuran (10 mL), 2M LDA tetrahydrofuran solution (5.2 mL) was slowly added dropwise at-78 ℃, after reaction for 30 minutes, DMF (189.67mg, 2.595 mmol) was slowly added dropwise, stirring was continued for 30 minutes, after completion of the reaction, an ammonium chloride aqueous solution was added to the reaction solution, ethyl acetate extraction was performed, the organic phase was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to obtain 6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-carboxaldehyde 5 (200 mg, white solid), yield: 84.62%.
1 H NMR(400MHz,CDCl3)δ10.36(d,J＝12.1Hz,1H),7.03(dt,J＝15.5,7.7Hz,1H),6.82–6.35(m,1H),4.39(m,2H),4.28(m,2H).
The fourth step
Preparation of (6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol
6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-carbaldehyde 5 (200mg, 1.098mmol) was dissolved in methanol (10 mL), sodium borohydride (29.08mg, 0.7686mmol) was slowly added at 0 ℃, after 1 hour of reaction, most of the methanol was removed, ammonium chloride aqueous solution was added, dichloromethane was extracted, the organic phase was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to give (6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (180 mg, white solid) in yield: 89.02%.
MS m/z(ESI):185.2(M+1)。
The fifth step
Preparation of 5- (bromomethyl) -6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine
(6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (180mg, 0.9774mmol) was dissolved in dichloromethane (10 mL), phosphine tribromide (264.88mg, 0.9774mmol) was added at 0 ℃, the reaction solution was stirred at room temperature for 1 hour, after the reaction was completed, the reaction solution was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether) to give 5- (bromomethyl) -6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin 7 (55 mg, white solid) with yield: 22.72 percent.
MS m/z(ESI):247.1(M+1)。
The sixth step
Preparation of 6-fluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxine
A solution of 5- (bromomethyl) -6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine 7 (52mg, 0.2105mmol), 4-fluoro-2-methoxy-5-nitrophenol 8 (47.27mg, 0.2526 mmol), potassium carbonate (58.19mg, 0.421mmol) and potassium iodide (3.49mg, 0.021mmol) in acetonitrile (5 mL) was stirred at 80 ℃ for 16 h. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 6-fluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin 9 (37 mg, yellow solid) in yield: 43.04 percent.
MS m/z(ESI):354.3(M+23)。
Seventh step
Preparation of 2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyaniline
6-fluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin 9 (30mg, 0.0849mmol), iron powder (14.22mg, 0.2547mmol), and ammonium chloride (9.08mg, 0.1698mmol) in ethanol: water =1:1 (6 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, quench with water and extract with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyaniline 10 (20 mg, brown oil), yield: 72.91%.
MS m/z(ESI):324.3(M+1)。
Eighth step
Preparation of 4- (3- (5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
A solution of 2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyaniline 10 (20mg, 0.0619mmol), 4- (phenoxycarbonyl) amino) thiophene-2,3 dimethyl dicarboxylate 11 (31.14mg, 0.0929mmol), and triethylamine (12.53mg, 0.1238mmol) in tetrahydrofuran (5 mL) was stirred at 25 ℃ for 16 hours. After the reaction was completed, water was added to quench, and extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 4- (3- (5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dimethyl dicarboxylate 12 (16 mg, yellow solid), yield: 45.72 percent.
MS m/z(ESI)：565.5(M+1)。
The ninth step
Preparation of 3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 12 (1695g, 0.0283mmol) and lithium hydroxide (2.03mg, 0.0849mmol) in tetrahydrofuran: water =1:1 (8 mL) was stirred at 25 ℃ for 1 hour. After the reaction was complete, water was added to quench the reaction, extracted with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was lyophilized by preparative purification to give 3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-127 (3.09 mg, yellow solid), yield: 21.20 percent.
MS m/z(ESI)：519.4(M+1)。
HPLC:98.41％(214nm),92.37％(254nm).
1 H NMR(400MHz,dmso)δ8.40(s,2H),7.13(d,J＝7.3Hz,1H),7.04(d,J＝11.4Hz,1H),6.94(dd,J＝9.0,5.7Hz,1H),6.75(t,J＝9.0Hz,1H),6.54(s,1H),4.89(s,2H),4.26(d,J＝23.5Hz,4H),3.78(s,3H).
Example 123
3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002201
First step of
Preparation of 2- (2-bromo-4,5-difluorophenoxy) ethyl-1-ol
After 2-bromo-4,5-difluorophenol 1 (5 g, 0.0239mol), 2-bromoethanol (5.97g, 0.0478mol) were dissolved in acetonitrile (50 mL), potassium carbonate (6.61g, 0.0478mol) and potassium iodide (3.97g, 0.0239mol) were added, the reaction was stirred at 80 ℃ for 16 hours, after the reaction was completed, the reaction was filtered with suction, the filtrate was concentrated, and the filtrate was purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 10/1) to obtain 2- (2-bromo-4,5-difluorophenoxy) ethyl-1-ol 3 (5 g, white solid) in yield: 82.68 percent.
1 H NMR(400MHz,CDCl 3 )δ7.39(dd,J＝9.4,8.4Hz,1H),6.80(dd,J＝11.6,6.9Hz,1H),4.09(dd,J＝5.1,3.8Hz,3H),3.99(dd,J＝5.1,3.7Hz,2H),2.38(s,1H).
Second step of
Preparation of 6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxins
To 2- (2-bromo-4,5-difluorophenoxy) ethyl-1-ol 3 (4g, 15.8mmol), palladium acetate (355mg, 1.58mmol), t BuDavePhos (1.08g, 3.16mmol), cesium carbonate (10.3g, 31.6mmol) and toluene (20 mL) are added, the reaction is carried out at 80 ℃ for 1 hour by microwave reaction, after the reaction is finished, the reaction solution is filtered, the filtrate is concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 20/1), and 6,7-difluoro-2,3-dihydrobenzo [ b ] is obtained][1,4]Dioxin 4 (1 g, white solid), yield: 36.79 percent.
1 H NMR(400MHz,CDCl 3 )δ6.61(t,J＝9.4Hz,1H),4.14(s,2H). 19 F NMR(376MHz,CDCl 3 )δ-146.12.
The third step
Preparation of 6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine-5-carbaldehyde
6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin 4 (1g, 5.81mmol) was dissolved in tetrahydrofuran (10 mL), 2M LDA tetrahydrofuran solution (5.8 mL) was slowly added dropwise at-78 ℃, after 30 minutes of reaction, DMF (0.51g, 6.96mmol) was slowly added dropwise, stirring was continued for 30 minutes, after completion of the reaction, an ammonium chloride aqueous solution was added to the reaction solution, ethyl acetate extraction was performed, the organic phase was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give 6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-formaldehyde 5 (1 g, white solid), yield: 86 percent.
1 H NMR(400MHz,CDCl 3 )δ10.35(s,1H),6.95(dd,J＝10.6,7.8Hz,1H),4.33(ddd,J＝7.8,6.0,3.6Hz,4H).
The fourth step
Preparation of (6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol
6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-carbaldehyde 5 (1 g, 4.7mmol) was dissolved in methanol (10 mL), sodium borohydride (378mg, 9.99mmol) was slowly added at 0 ℃, after 1 hour of reaction, most of the methanol was removed by rotation, ammonium chloride aqueous solution was added, dichloromethane was extracted, the organic phase was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 3/1) to give (6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (900 mg, white solid) in yield: 94.72 percent.
MS m/z(ESI):185.0(M-17)。
The fifth step
Preparation of 5- (bromomethyl) -6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxine
(6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (900mg, 4.45mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (1.4 g, 5.34mmol) was added, carbon tetrabromide (1.77g, 5.34mmol) was added after stirring for a while, the reaction solution was stirred at room temperature for 1 hour, after the completion of the reaction, the reaction solution was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether) to obtain 5- (bromomethyl) -6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin 7 (800 mg, white solid) with a yield: 67.82%.
1 H NMR(400MHz,CDCl3)δ6.70(dd,J＝10.7,8.0Hz,1H),4.52(s,2H),4.29(ddd,J＝7.4,5.9,3.5Hz,4H).
The sixth step
Preparation of 6,7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxine
A solution of 5- (bromomethyl) -6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin 7 (300mg, 1.13mmol), 4-fluoro-2-methoxy-5-nitrophenol 8 (211mg, 1.13mmol), potassium carbonate (312mg, 2.26mmol), and potassium iodide (93mg, 0.56mmol) in acetonitrile (5 mL) was stirred at 80 ℃ for 16 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 5/1) to give preparation 9 (320 mg, yellow solid) of 6,7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin in yield: 68.53%.
MS m/z(ESI):394.0(M+23)。
Seventh step
Preparation of 5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline
Preparation of 6,7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzo [ b ] [1,4] dioxin 9 (340mg, 0.91mmol), iron powder (153mg, 2.74mmol), and ammonium chloride (146mg, 2.74mmol) in ethanol: water =1:1 (6 mL) was stirred at 80 ℃ for 1 hour. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: dichloromethane/methanol = 10/1) to give 5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 10 (180 mg, brown oil) in yield: 51.83%.
MS m/z(ESI):342.0(M+1)。
Eighth step
Preparation of dimethyl 4- (3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
Preparation of 5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 10 (250mg, 0.73mmol), 4- (phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester 11 (294mg, 0.87mmol) and triethylamine (148mg, 1.46mmol) in tetrahydrofuran (5 mL) was stirred at 25 ℃ for 16 hours. After the reaction was completed, water was added to quench, and extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate = 2/1) to obtain 4- (3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 12 (580 mg, yellow solid), yield: 95.15%.
MS m/z(ESI)：583.0(M+1)。
The ninth step
Preparation of 3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl- (3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate 12 (520mg, 0.89mmol) and lithium hydroxide (64mg, 2.67mmol) in tetrahydrofuran: water =1:1 (8 mL) was stirred at 25 ℃ for 1 hour. After the reaction was completed, water was added to quench after the reaction was completed, extraction was performed with ethyl acetate (3 × 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was lyophilized by preparative purification to give 3- (5- ((6,7-difluoro-2,3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-128 (119.7 mg, yellow solid), yield: 24.03 percent.
MS m/z(ESI)：537.0(M+1)。
HPLC:96.11％(214nm),100％(254nm).
1 H NMR(400MHz,DMSO-d6)δ14.53(s,1H),11.97(s,1H),7.33(s,1H),7.23(d,J＝7.3Hz,1H),7.13(d,J＝8.1Hz,1H),7.11(d,J＝7.7Hz,1H),4.95(s,2H),4.25(s,4H),3.82(s,3H).
Example 124
3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002231
First step of
Preparation of 1-bromo-4-chloro-2- (2,2-diethoxyethoxy) benzene
2-bromo-5-chlorophenol 216a (10g, 48.2mmol) was dissolved in DMF (100 mL), and then 2-bromo-1,1-diethoxyethane (9.5g, 48.2mmol) and potassium carbonate (13.3g, 96.4mmol) were added, and stirred at 100 ℃ for 16 hours, after the reaction was completed, the reaction solution was filtered, washed with water, extracted with ethyl acetate, and dried and spin-dried by silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain 1-bromo-4-chloro-2- (2,2-diethoxyethoxy) benzene 216b (11 g, white solid) with a yield of 70.5%.
MS m/z(ESI)：347.1(M+Na)。
Second step of
Preparation of 7-bromo-4-chlorobenzofuran
1-bromo-4-chloro-2- (2,2-diethoxyethoxy) benzene 216b (11g, 34.0mmol) was dissolved in toluene (100 mL), then PPA (11.5g, 34.0mmol) was added, stirring was performed at 80 ℃ for 2 hours, after the reaction was completed, ice water was added to the reaction solution, extraction was performed with ethyl acetate, and organic phase was dried and spin-dried and was separated and purified by a silica gel column (petroleum ether) to give 7-bromo-4-chlorobenzofuran 216c (5.7 g, white solid) in a yield of 72.4%.
1 H NMR(400MHz,CDCl 3 )δ7.72(d,J＝2.0Hz,1H),7.39(d,J＝8.3Hz,1H),7.14(d,J＝8.3Hz,1H),6.94(d,J＝2.1Hz,1H).
The third step
Preparation of 7-bromo-4-chloro-2,3-dihydrobenzofuran
7-bromo-4-chlorobenzofuran 216C (5.7 g,24.6 mmol) was dissolved in ethanol (100 mL), rh/C (520mg, 4.92mmol) was added, and the mixture was stirred under pressure with hydrogen at 25 ℃ for 16 hours, after the reaction was completed, the reaction mixture was filtered, and the filtrate was separated and purified by silica gel column (petroleum ether) to give 7-bromo-4-chloro-2,3-dihydrobenzofuran 216d (5.1 g, white solid) in 88.8% yield.
1 H NMR(400MHz,CDCl 3 )δ7.19(d,J＝8.5Hz,1H),6.72(d,J＝8.5Hz,1H),4.70(t,J＝8.8Hz,2H),3.33(t,J＝8.8Hz,2H).
The fourth step
Preparation of methyl 4-chloro-2,3-dihydrobenzofuran-7-carboxylate
7-bromo-4-chloro-2,3-dihydrobenzofuran 216d (2g, 8.6 mmol) was dissolved in methanol (100 mL), then Pd (dppf) Cl2 (0.63g, 0.86mmol) and potassium acetate (1.69g, 17.2 mmol) were added, and after completion of the reaction, the reaction solution was filtered, and the filtrate was isolated and purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to give 4-chloro-2,3-dihydrobenzofuran-7-carboxylic acid methyl ester 216e (650 mg, white solid) in 35.5% yield.
MS m/z(ESI)：213.1(M+H)。
The fifth step
Preparation of (4-chloro-2,3-dihydrobenzofuran-7-yl) methanol
4-chloro-2,3-dihydrobenzofuran-7-carboxylic acid methyl ester 216e (650mg, 3.1mmol) was dissolved in tetrahydrofuran (10 mL), lithium aluminum hydrogen (235mg, 6.2mmol) was added at 0 ℃ and stirred for 1 hour, after the reaction was completed, an ammonium chloride aqueous solution was added to the reaction solution, ethyl acetate was extracted, and organic phase was dried and spin-dried and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 5/1) to obtain (4-chloro-2,3-dihydrobenzofuran-7-yl) methanol 216f (400 mg, white solid) in 69.9% yield.
MS m/z(ESI)：167.1(M-17)。
The sixth step
Preparation of 7- (bromomethyl) -4-chloro-2,3-dihydrobenzofuran
(4-chloro-2,3-dihydrobenzofuran-7-yl) methanol 216f (400mg, 2.2mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (682mg, 2.6 mmol) was added at 0 ℃ and stirred for 10 minutes, carbon tetrabromide (862mg, 2.6 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was spin-dried and separated and purified by silica gel column (petroleum ether) to obtain 216g (380 mg, white solid) of 7- (bromomethyl) -4-chloro-2,3-dihydrobenzofuran in 69.8% yield.
1 H NMR(400MHz,CDCl 3 )δ7.06(d,J＝8.2Hz,1H),6.81(d,J＝8.2Hz,1H),4.70(t,J＝8.8Hz,2H),4.45(s,2H),3.26(t,J＝8.8Hz,2H).
Seventh step
Preparation of 4-chloro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran
7- (bromomethyl) -4-chloro-2,3-dihydrobenzofuran 216g (380mg, 1.5 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (287mg, 1.5 mmol) were dissolved in acetonitrile (10 mL), followed by addition of potassium carbonate (424mg, 3.1mmol), potassium iodide (255mg, 1.5 mmol), reaction liquid was stirred at 70 ℃ for 16 hours, after completion of the reaction, water was added to the reaction liquid, ethyl acetate was extracted, organic phase was dried and concentrated, and then purified by silica gel column (petroleum ether/ethyl acetate = 5/1) to give 4-chloro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran 216h (120 mg, yellow solid) with a yield of 22.6%.
MS m/z(ESI)：376.0(M+Na)。
Eighth step
Preparation of 5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyaniline
4-chloro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran 216h (120mg, 0.34mmol) was dissolved in ethanol/ammonium chloride aqueous solution =4/1 (10 mL), iron powder (189mg, 3.4 mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyaniline 216i (100 mg, yellow oily liquid) in 90.9% yield.
MS m/z(ESI)：324.1(M+H)。
The ninth step
Preparation of 4- (3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester
5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyaniline 216i (100mg, 0.31mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (114mg, 0.34mmol) and triethylamine (94mg, 0.93mmol), the reaction solution was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction solution was concentrated, and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4- (3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 216j (150 mg, yellow solid) with a yield of 85.6%.
MS m/z(ESI)：565.0(M+H)。
The tenth step
Preparation of 3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 216j (150mg, 0.26mmol) was dissolved in tetrahydrofuran/water =1/1 (10 mL), lithium hydroxide (19mg, 0.78mmol) was added, the reaction solution was stirred at room temperature for 1 hour, and after completion of the reaction, drying was carried out to prepare 3- (5- ((4-chloro-2,3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-129 (55 mg, yellow solid) in 40.8% yield.
MS m/z(ESI)：519.1(M+H)。
HPLC:97.95％(214nm),95.03％(254nm)。
1 H NMR(400MHz,DMSO)δ11.84(s,1H),7.23(m,3H),7.11(d,J＝11.5Hz,1H),6.92(d,J＝8.2Hz,1H),4.86(s,2H),4.62(t,J＝8.8Hz,2H),3.82(s,3H),3.24(t,J＝8.7Hz,2H).
Example 125
3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
Figure BDA0003608368340002251
First step of
Preparation of 2-bromo-1- (2,2-diethoxyethoxy) -3-fluorobenzene
A solution of 2-bromo-3-fluorophenol 217a (10g, 0.05mol) in DMF (30 mL) was added dropwise to sodium hydride, and the reaction mixture was stirred at 25 ℃ for 1 hour. Subsequently, 2-bromo-1,1-dioxyethane (11.36g, 0.05mol) was added to the reaction mixture, and stirred at 153 ℃ for 2 hours, after the reaction was completed, the reaction solution was suction-filtered, and after the filtrate was concentrated, the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 19/1) to obtain 2-bromo-1- (2,2-diethoxyethoxy) -3-fluorobenzene 217b (9.4 g, colorless liquid), yield: 52.5 percent.
1 H NMR(400MHz,DMSO)δ7.37(d,J＝6.8Hz,1H),7.02-6.95(m,2H),4.86-4.83(m,1H),4.07(d,J＝5.2Hz,2H),3.74–3.69(m,2H),3.65–3.61(m,2H),1.18-1.12(m,12H).
Second step of
Preparation of 7-bromo-6-fluorobenzofuran
To 2-bromo-1- (2,2-diethoxyethoxy) -3-fluorobenzene 217b (1.4 g,4.6 mmol) and polyphosphoric acid (1.55g, 4.6 mmol) was added chlorobenzene (20 mL) and the reaction mixture was stirred at 80 ℃ for 2 hours. After the reaction was complete, water was added and quenched and extracted with ethyl acetate (3 × 10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to give 7-bromo-6-fluorobenzofuran 217c (680 mg, white solid) in yield: 60.9 percent.
1 H NMR(400MHz,DMSO)δ8.17(d,J＝2.0Hz,1H),7.73-7.69(m,1H),7.34-7.30(m,1H),7.13(d,J＝2.0Hz,1H).
The third step
Preparation of 7-bromo-6-fluoro-2,3-dihydrobenzofuran
7-bromo-6-fluorobenzofuran 217C (2.6g, 12.1mmol) was dissolved in ethanol (50 mL), rh/C (620mg, 6.1mmol) was added, and the reaction mixture was stirred under pressure with hydrogen at 25 ℃ for 16 hours, after completion of the reaction, the reaction mixture was filtered, and the filtrate was isolated and purified by silica gel column (petroleum ether) to give 7-bromo-6-fluoro-2,3-dihydrobenzofuran 217d (1.9 g, white solid) in 72.4% yield.
1 H NMR(400MHz,CDCl 3 )δ7.06–7.00(m,1H),6.64–6.57(m,1H),4.72(t,J＝8.8Hz,2H),3.33–3.24(m,2H).
The fourth step
Preparation of 6-fluoro-2,3-dihydrobenzofuran-7-carbaldehyde
7-bromo-6-fluoro-2,3-dihydrobenzofuran 217d (1.25g, 5.8mmol) was dissolved in tetrahydrofuran (20 mL), isopropyl magnesium chloride (420mg, 2.9mmol) was added at 0 deg.C, stirring was carried out for 30 minutes, then n-butyl lithium (560mg, 8.7mmol) was added, stirring was carried out for 30 minutes, and finally DMF (470mg, 6.4mmol) was added, stirring was carried out for 30 minutes, after completion of the reaction, saturated ammonium chloride was added, ethyl acetate was extracted, and the organic phase was purified by silica gel column (petroleum ether/ethyl acetate = 5/1) separation to give 6-fluoro-2,3-dihydrobenzofuran-7-carbaldehyde 217e (700 mg, white solid) in 72.7% yield.
MS m/z(ESI)：167.1(M+H)。
The fifth step
Preparation of (6-fluoro-2,3-dihydrobenzofuran-7-yl) methanol
6-fluoro-2,3-dihydrobenzofuran-7-carbaldehyde 217e (700mg, 4.2mmol) is dissolved in tetrahydrofuran (10 mL), lithium aluminum hydrogen (320mg, 8.2mmol) is added at 0 ℃ and stirred for 1 hour, after the reaction is finished, an ammonium chloride aqueous solution is added into the reaction liquid, ethyl acetate is extracted, and organic phase drying and spin drying are carried out, and separation and purification are carried out through a silica gel column (petroleum ether/ethyl acetate = 3/1), so that (6-fluoro-2,3-dihydrobenzofuran-7-yl) methanol 217f (400 mg, white solid) is obtained, and the yield is 56.6%.
MS m/z(ESI)：151.1(M-17)。
The sixth step
Preparation of 7- (bromomethyl) -6-fluoro-2,3-dihydrobenzofuran
(6-fluoro-2,3-dihydrobenzofuran-7-yl) methanol 217f (400mg, 2.4mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (749mg, 2.8mmol) was added at 0 ℃ and the mixture was stirred for 10 minutes, followed by addition of carbon tetrabromide (946mg, 2.8mmol), and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was spin-dried and separated and purified by silica gel column (petroleum ether) to obtain 217g (250 mg, white solid) of 7- (bromomethyl) -6-fluoro-2,3-dihydrobenzofuran in a yield of 45.1%.
1 H NMR(400MHz,CDCl 3 )δ7.13–6.98(m,1H),6.55(dd,J＝9.7,8.4Hz,1H),4.71(t,J＝8.7Hz,2H),4.51(s,2H),3.19(t,J＝8.7Hz,2H).
Seventh step
Preparation of 6-fluoro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran
4-fluoro-2-methoxy-5-nitrophenol (206mg, 1.1mmol) was dissolved in DMF (3 mL), followed by addition of sodium hydrogen (32mg, 1.3mmol) at 0 ℃, stirring of the reaction solution for 1 hour, followed by addition of preparation 217g (250mg, 1.1mmol) of 7- (bromomethyl) -6-fluoro-2,3-dihydrobenzofuran, stirring of the reaction solution at 0 ℃ for 1 hour, after completion of the reaction, addition of water and extraction of ethyl acetate to the reaction solution, drying and concentration of the organic phase, and separation and purification with a silica gel column (petroleum ether/ethyl acetate = 5/1) to give 6-fluoro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran 217h (150 mg, yellow solid) in a yield of 40.4%.
MS m/z(ESI)：360.0(M+Na)。
Eighth step
Preparation of 2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyaniline
6-fluoro-7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2,3-dihydrobenzofuran 217h (150mg, 0.44mmol) was dissolved in ethanol/ammonium chloride aqueous solution =4/1 (10 mL), iron powder (246mg, 4.4 mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated and separated and purified with a silica gel column (petroleum ether/ethyl acetate = 3/1) to obtain 2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyaniline 217i (120 mg, yellow oily liquid) in 88.8% yield.
MS m/z(ESI)：308.2(M+H)。
The ninth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylate
2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyaniline 217i (120mg, 0.39mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2,3-dicarboxylic acid dimethyl ester (144mg, 0.43mmol) and triethylamine (118mg, 1.17mmol), the reaction solution was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction solution was concentrated, and separated and purified by a silica gel column (petroleum ether/ethyl acetate = 3/1) to give 4- (3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 217j (180 mg, yellow solid) in 84.2% yield.
MS m/z(ESI)：549.1(M+H)。
The tenth step
Preparation of 3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid
4- (3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2,3-dicarboxylic acid dimethyl ester 217j (180mg, 0.33mmol) was dissolved in tetrahydrofuran/water =1/1 (10 mL), lithium hydroxide (41mg, 0.99mmol) was added, the reaction solution was stirred at room temperature for 1 hour, and after completion of the reaction, drying was performed to prepare 3- (2-fluoro-5- ((6-fluoro-2,3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) -2,4-dioxo-1,2,3,4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-130 (84 mg, yellow solid) with a yield of 50.7%.
MS m/z(ESI)：503.0(M+H)。
HPLC:100％(214nm),100％(254nm)。
1 H NMR(400MHz,DMSO)δ12.01(s,1H),7.37(s,1H),7.29–7.23(m,2H),7.12(d,J＝11.6Hz,1H),6.69(dd,J＝10.1,8.2Hz,1H),4.87(s,2H),4.62(t,J＝8.7Hz,2H),3.81(s,3H),3.18(t,J＝8.7Hz,3H).
Biological evaluation
Test example 1 measurement of human GnRHR Activity of Compounds of the present invention
The method is used for measuring the antagonism of the activity of the human GnRHR protein expressed in the human GnRHR/HEK293 stable cell strain by the compound.
1. Test materials and apparatus
1.1 culture Medium
F12(Gibco,Cat#11765-047)；FBS(Corning,Cat#35-076-CV)；Geneticin(Invitrogen,Cat#10131)；Penicillin/Streptomycin(Invitrogen,Cat#15140)。
1.2 reagents
Fluo-4 Direct(Invitrogen,Cat#F10471)；HBSS(Gibco,Cat#14025076)；HEPES(Gibco,Cat#15630080)；Bonine Serum Albumin(Sigma,Cat#B2064-100G)。
1.3 consumable for instruments
384well Poly-D-Lysine protein coating plate(Greiner,Cat#781946)；FLIPR(Molecular Devices)；Vi-cell XR Cell Viability Analyzer(Beckman Coulter)；Incubator(Thermo)。
2. Experimental procedure
2.1 inoculation of human GnRHR/HEK293 stably transfected cell lines in 384-well cell culture plates at 12000 cells/well/25. Mu.l, 5% CO at 37 ℃% 2 Culturing overnight;
2.2 Freeze-thawing 20X Component A to room temperature, diluting the solution to 2X working concentration by using Assay Buffer, and standing at room temperature for later use;
2.3 equilibrating the cell culture plate at room temperature for 10 minutes, removing the medium, adding 20. Mu.L of Assay Buffer and 20. Mu.L of 2X Component A,200g, RT centrifuge 3-5sec,37 ℃ and incubating for 2 hours;
2.4 Compounds were diluted 3-fold in 384PP _DMSOplates using DMSO, and then transferred 240nl/well each to working plates using Echo 550, 200g, RT,1min; adding 40 μ l Assay Buffer into the working plate, after 200g, RT and 1min, placing in an oscillator 2500rpm,20min, mixing uniformly, and then 200g, RT and 1min for standby;
2.5 preparing 180nM LH-RH (6X) by using Assay Buffer, taking 50ul to 3657 plates for later use;
2.6 taking out the cell culture plate, standing for 10min at room temperature, adding 10 μ l of the compound diluted in the step 2.4 into the corresponding hole, and standing for 30min at 25 ℃;
2.7 mu.l of the compound diluted in step 2.5 was added to the corresponding experimental wells using FLIPR Tetra and the data collected.
3. Experimental data:
the antagonistic activity of the human GnRHR compound is measured by the experiment, and the measured IC 50 The values are shown in Table 1.
TABLE 1 IC of the human GnRHR antagonistic Activity of the Compounds of the present invention 50 Value of
Figure BDA0003608368340002281
Figure BDA0003608368340002291
4. And (4) experimental conclusion:
the data show that the compound has obvious antagonism on the activity of human GnRHR.
Test example 2 hERG inhibition test of the Compound of the present invention
Human ventricular rapid activation delayed rectifier potassium current (IKr) is mainly mediated by the human ether-a-go-go related gene (hERG) potassium channel in the heart. Inhibition of IKr is the most common mechanism by which non-cardiac drugs cause prolonged ventricular muscle action potential. Prolonged action potential duration will lead to prolonged QT interval on clinical electrocardiogram, which is associated with dangerous ventricular arrhythmias, torsion tachycardia. Because of these cardiotoxic effects, many drugs have been withdrawn from late clinical trials and it is important to identify inhibitors early in drug development. Therefore, studying the effect of drugs on the hERG potassium channel in a system of heterologous expression is a non-clinical study recommended by ICH as a test drug for the effect on QT interval.
The hERG inhibition study was conducted to quantify the in vitro effect of the compounds of the invention on potassium-selective IKr currents generated under normoxic conditions in stably transfected HEK 293 cells harboring the human ether-a-go-go related gene (hERG).
The cell seed was placed on an inverted microscope stage in a cell well and one cell in the well was randomly selected for testing. The perfusion system was fixed on an inverted microscope stage and cells were perfused continuously with ECS.
A manual patch clamp test recording microelectrode was prepared with a capillary glass tube, into which the intracellular fluid was filled. On the day of the patch clamp test, electrodes were prepared using borosilicate glass tubes (GC 150TF-10, harvard Apparatus Co. UK). The resistance after the electrode is filled with ICS is between 2 and 5M omega.
The clamp voltage is-80 mV, the first depolarization to +60mV and maintain 850ms open hERG channel. The voltage was then set to-50 mV and maintained for 1275ms, producing a bounce current or called tail current, the peak of which was measured and used for analysis. Eventually, the voltage returns to the clamped voltage (-80 mV). This program of command voltages was repeated every 15s interval during the test.
And in the beginning stage of recording the perfusion of the solvent contrast working solution, monitoring the tail current peak value until more than 3 scanning curves are stabilized, and perfusing the compound working solution to be tested until the inhibition effect of the compound working solution on the hERG current peak value reaches a stable state. Generally, the peak value of the nearest continuous 3 current curves is basically coincided to be used as a standard for judging whether the stable state exists or not, and a compound with a certain concentration is continuously poured after the stable state is achieved.
All percent cytostatic, IC, values recorded for each concentration were averaged 50 Values are derived from the concentration effect curves. IC (integrated circuit) 50 All compounds with values above 10 μ M are considered not potent inhibitors of the hERG channel, but have an IC below 1 μ M 50 Compounds of value are considered potent hERG channel inhibitors.
Experimental data:
TABLE 2 IC of the Compounds of the invention in the hERG inhibition assay 50 The value is obtained.
Figure BDA0003608368340002301
Figure BDA0003608368340002311
And (4) experimental conclusion:
the above data show that none of the compounds of the present invention are potent inhibitors of the hERG channel.
Test example 3 rat pharmacokinetic testing of the Compound of the invention
1. Purpose of the experiment:
SD rats are used as test animals, and the drug concentration in plasma of the rats at different times after the administration of the compound of the embodiment through gastric gavage is determined by an LC/MS/MS method. The pharmacokinetic behavior of the compounds of the invention in rats was studied and their pharmacokinetic profile was evaluated.
2. Experimental protocol
2.1 Experimental drugs
The compound of the embodiment of the invention is prepared by self.
2.2 Experimental animals
Healthy adult SD rats.
2.3 pharmaceutical formulation
Weighing appropriate amount of sample, gavage to deliver the solvent formulation 2% DMA +97.4% (0.5% CMC-Na) +0.6%2M NaOH, ultrasound to make up a suspension of 0.3 mg/mL.
2.4 administration
SD rats are respectively gavaged after being fasted overnight, and the administration dosage is 3mg/kg.
3. The experimental steps are as follows:
the gavage administration group collected blood at 0.1mL before and 30min, 1h, 4h, 6h, 8h, 12h, 24h, 48h, 72h, 96h, 168h, and 240h after administration, placed in heparinized tubes, centrifuged at 3500rpm for 10 minutes, separated plasma, and stored at-20 ℃. Food/free water was returned 4 hours after dosing.
The LC/MS/MS method is used for measuring the content of the compound to be tested in the plasma of rats after the gavage administration.
4. Experimental data:
table 3 pharmacokinetic parameters of the compounds of the invention.
Figure BDA0003608368340002312
Figure BDA0003608368340002321
5. And (4) experimental conclusion:
the data show that the compound has good drug absorption in the rat stomach, obviously prolonged half life and obvious drug absorption effect.
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, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (11)

1. A compound, its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound of formula I:
Figure FDA0003608368330000011
Wherein Y is selected from the following groups, unsubstituted or substituted by X and/or-O-J: c 6-20 An aromatic or 5-20 membered heteroaromatic ring; preferably, A and-E-G are in the meta position, X and-O-J are in the meta position, A and X are in the ortho position, -E-G and-O-J are in the ortho position;
a is selected from unsubstituted or optionally substituted by one, two or more R 1 Substituted by
Figure FDA0003608368330000012
B is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted 5-20 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-20 membered heterocyclic ring or 5-20 membered heteroaromatic ring;
e is selected from O, C (O), NH, NC 1-40 Alkyl, N (C) 3-20 Cycloalkyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、NHC(O)、OCH 2 、O(CH 2 ) 2 、NHCH 2 Or N (C) 1-40 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-40 Alkyl radical, C 6-20 Aryl, 5-20 membered heterocyclyl or 5-20 membered heteroaryl;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-40 Alkyl radical, C 3-20 Cycloalkyl radical, C 6-20 Aryl, 5-20 membered heteroaryl, orJ-O-is fused to the Y to which it is attached, preferably to the phenyl ring to which it is attached, to form a 5-20 membered heteroaromatic ring;
x is selected from hydrogen or halogen;
Figure FDA0003608368330000013
is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl group, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 、COOH、-COOM、NH-C(O)-NH 2 、-C(O)C 1-40 Alkyl, -C (O) OC 1-40 Alkyl, -C (O) C 1-40 Hydroxyalkyl, -S (O) 2 -NHC 1-40 Alkyl, -S (O) 2 C 6-20 Aryl, -S (O) 2 -5-20 membered heteroaryl, NH-C (O) -NH-OC 1-40 Alkyl or NH 2 -C (O) -; wherein M is selected from cations;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl group, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyl oxygen radicalBase, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 COOH, NH-C (O) -NH or NH-C (O) -NH-OC 1-40 An alkyl group;
each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-40 Alkyl radical, C 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl group, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy or NH 2 ；
Preferably, the structure of the compound of formula I is as shown in formula I':
Figure FDA0003608368330000021
wherein A is selected from unsubstituted or optionally substituted with one, two or more R 1 Substituted
Figure FDA0003608368330000022
B is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted 5-20 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-20 membered heterocyclic ring or 5-20 membered heteroaromatic ring;
e is selected from O, C (O), NH, NC 1-40 Alkyl, S (O) 2 、CH 2 、NHS(O) 2 、NHC(O)、OCH 2 、O(CH 2 ) 2 、NHCH 2 Or N (C) 1-40 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-40 Alkyl radical, C 6-20 Aryl, 5-20 membered heterocyclyl or 5-20 membered heteroaryl;
j is selected from unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-40 Alkyl radical, C 6-20 Aryl, 5-20 membered heteroaryl, or J-O-fused with the phenyl ring to which it is attached to form a 5-20 membered heteroaryl ring;
x is selected from hydrogen or halogen;
Figure FDA0003608368330000023
Is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl group, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyloxy radical, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 、COOH、NH-C(O)-NH、-C(O)C 1-40 Alkyl, -C (O) C 1-40 Hydroxyalkyl, -S (O) 2 -NHC 1-40 Alkyl, -S (O) 2 C 6-20 Aryl, -S (O) 2 -5-20 membered heteroaryl or NH-C (O) -NH-OC 1-40 An alkyl group;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy, NH 2 COOH, NH-C (O) -NH or NH-C (O) -NH-OC 1-40 An alkyl group;
each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-40 Alkyl radical, C 1-40 Alkyl radical, C 2-40 Alkenyl radical, C 2-40 Alkynyl, C 3-40 Cycloalkyl radical, C 3-40 Cycloalkenyl radical, C 3-40 Cycloalkynyl, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy radical, C 2-40 Alkynyloxy, C 3-40 Cycloalkyloxy radical, C 3-40 Cycloalkenyloxy, C 3-40 Cycloalkynyloxy, C 6-20 Aryloxy, 5-20 membered heteroaryloxy, 3-20 membered heterocyclyloxy or NH 2 。
2. A compound of formula I according to claim 1, wherein Y is selected from the following groups, unsubstituted or substituted by X and/or-O-J: a benzene ring or a pyridine ring;
b is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted 5-14 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-14 membered heterocyclic ring or 5-14 membered heteroaromatic ring;
e is selected from O, C (O), NH, NC 1-8 Alkyl, N (C) 3-8 Cycloalkyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、NHC(O)、OCH 2 、O(CH 2 ) 2 、NHCH 2 Or N (C) 1-8 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl radical, C 6-14 Aryl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure FDA0003608368330000031
And B is selected from the thiophene ring, G is selected from unsubstituted or optionally substituted with one, two or more R 4 Substituted C 9-14 Aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl, said aryl, heteroaryl or heterocyclyl preferably being fused ring groups;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl radical, C 3-8 Cycloalkyl radical, C 6-14 Aryl, 5-14 membered heteroaryl, or J-O-is attached to C at the ortho position of the phenyl ring to which it is attached to form a 5-14 membered heteroaryl ring which is attached in juxtaposition to the phenyl ring;
x is selected from hydrogen or halogen;
Figure FDA0003608368330000033
is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 、COOH、-COOM、NH-C(O)-NH 2 、-C(O)C 1-8 Alkyl, -C (O) OC 1-8 Alkyl, aryl, heteroaryl, and heteroaryl,-C(O)C 1-8 Hydroxyalkyl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl, NH-C (O) -NH-OC 1-8 Alkyl or NH 2 -C (O) -; wherein M is selected from monovalent cations;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyloxy radical, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 COOH, NH-C (O) -NH or NH-C (O) -NH-OC 1-8 An alkyl group.
Each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-8 Alkyl radical, C 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy or NH 2 ；
Preferably, B is selected from unsubstituted or optionally substituted with one, two or more R 2 A substituted 5-14 membered heteroaromatic ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-14 membered heterocyclic ring or 5-14 membered heteroaromatic ring;
e selectionFrom O, C (O), NH, NC 1-8 Alkyl, S (O) 2 、CH 2 、NHS(O) 2 、NHC(O)、OCH 2 、O(CH 2 ) 2 、NHCH 2 Or N (C) 1-8 Alkyl) CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure FDA0003608368330000032
And B is selected from the thiophene ring, G is selected from unsubstituted or optionally substituted with one, two or more R 4 Substituted C 9-14 Aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl, preferably fused ring groups;
j is selected from unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl radical, C 6-14 Aryl, 5-14 membered heteroaryl, or J-O-is attached to the C at the ortho position of the phenyl ring to which it is attached to form a 5-14 membered heteroaromatic ring which is attached to the phenyl ring in juxtaposition;
x is selected from hydrogen or halogen;
Figure FDA0003608368330000034
is a linking site;
each R 1 、R 2 、R 3 、R 4 、R 5 Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 An aryloxy group,5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 、COOH、NH-C(O)-NH、-C(O)C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl or NH-C (O) -NH-OC 1-8 An alkyl group;
each R a Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy, NH 2 COOH, NH-C (O) -NH or NH-C (O) -NH-OC 1-8 An alkyl group.
Each R b Identical or different, independently of one another, from hydrogen, halogen, OH, CN, NO 2 Oxo (= O), COOH, COOC 1-8 Alkyl radical, C 1-8 Alkyl radical, C 2-8 Alkenyl radical, C 2-8 Alkynyl, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkenyl radical, C 3-8 Cycloalkynyl group, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy radical, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyloxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy, 5-14 membered heteroaryloxy, 3-8 membered heterocyclyloxy or NH 2 。
3. The compound of formula I, its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound according to claim 1 or 2, characterized in that,
b is selected from unsubstituted or optionally substituted by one, two or more R 2 A substituted thiophene, furan, imidazole, or pyrazole ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 Substituted thiophene, furan, imidazole, pyrazole, pyrrole, tetrahydropyrrole rings;
e is selected from O, C (O), NH, NCH 3 N (-cyclopropyl) CH 2 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 、NHC(O)、NHCH 2 Or N (CH) 3 )CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure FDA0003608368330000041
And B is selected from the thiophene ring, G is selected from unsubstituted or optionally substituted with one, two or more R 4 Substituted C 9-14 Aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl, which aryl, heteroaryl or heterocyclyl may preferably be fused ring groups;
j is selected from H, unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl radical, C 3-8 Cycloalkyl or J-O-is connected with C at ortho position in the benzene ring connected with the cycloalkyl or J-O-to form an oxazole ring, a thiazole ring, an isoxazole ring, a pyrrole ring, a furan ring, an imidazole ring, a pyridine ring or a pyrazine ring which is connected with the benzene ring in parallel; preferably, J is selected from H, methyl, cyclopropylmethyl, n-butyl, methoxyethyl, cyclopentyl;
x is selected from hydrogen, fluorine, chlorine or bromine;
Figure FDA0003608368330000044
is a linking site;
each R 1 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOM, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C (O) C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, ph-NH-C (O) -NH-OC 1-8 Alkyl radical, C 1-8 alkyl-NH-C (O) -, di (C) 1-8 Alkyl) N-C 1-8 Alkyl, hydroxy C 1-8 An alkyl group; m is selected from monovalent cations, e.g. Na + 、K + Or
Figure FDA0003608368330000042
Each R 2 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOM, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C (O) C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl, ph-NH-C (O) -NH-OC 1-8 Alkyl radical, C 1-8 alkyl-NH-C (O) -, di (C) 1-8 Alkyl) N-C 1-8 Alkyl, hydroxy C 1-8 An alkyl group; m is selected from monovalent cations, e.g. Na + 、K + Or
Figure FDA0003608368330000043
Each R 3 Are identical or different and are each independently selected from hydrogen, oxo (= O), C 1-8 Alkyl, COOH or COOC 1-8 An alkyl group;
each R 4 Identical or different, independently of one another, from hydrogen, halogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl or C 1-8 An alkyloxy group;
each R 5 Identical or different, independently of one another, from hydrogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl or C 1-8 An alkyloxy group;
each R a Are identical or different and are selected independently of one anotherHydrogen, halogen, C 1-8 Alkyl radical, C 3-8 Cycloalkyl or C 1-8 An alkyloxy group;
preferably, B is selected from unsubstituted or optionally substituted with one, two or more R 2 A substituted thiophene, furan, imidazole, or pyrazole ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 Substituted thiophene, furan, imidazole, pyrazole, pyrrole, tetrahydropyrrole rings;
e is selected from O, C (O), NH, NCH 3 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 、NHC(O)、NHCH 2 Or N (CH) 3 )CH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted C 1-8 Alkyl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected from
Figure FDA0003608368330000051
And B is selected from the thiophene ring, G is selected from unsubstituted or optionally substituted with one, two or more R 4 Substituted C 9-14 Aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl, which aryl, heteroaryl or heterocyclyl may preferably be fused ring groups;
j is selected from unsubstituted or optionally substituted by one, two or more R 5 Substituted C 1-8 Alkyl or J-O-is connected with C at ortho position in the benzene ring connected with the alkyl or J-O-to form oxazole ring, thiazole ring, isoxazole ring, pyrrole ring, furan ring, imidazole ring, pyridine ring or pyrazine ring which is connected with the benzene ring in parallel;
x is selected from hydrogen, fluorine, chlorine or bromine;
Figure FDA0003608368330000053
is a linking site;
each R 1 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C(O)C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl or Ph-NH-C (O) -NH-OC 1-8 An alkyl group;
each R 2 Identical or different, independently of one another, from hydrogen, C 1-8 Alkyl, COOH, COOC 1-8 Alkyl, NH-C (O) -NH-OC 1-8 Alkyl, -C (O) C 1-8 Alkyl, -C (O) C 1-8 Hydroxyalkyl or Ph-NH-C (O) -NH-OC 1-8 An alkyl group;
each R 3 Are identical or different and are each independently selected from hydrogen, oxo (= O), C 1-8 Alkyl, COOH or COOC 1-8 An alkyl group;
each R 4 Identical or different, independently of one another, from hydrogen, halogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl, -S (O) 2 -NHC 1-8 Alkyl, -S (O) 2 C 6-14 Aryl, -S (O) 2 -5-14 membered heteroaryl or C 1-8 An alkyloxy group;
each R 5 Identical or different, independently of one another, from hydrogen, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-8 Alkyl, 5-14 membered heteroaryl or C 1-8 An alkyloxy group;
each R a Identical or different, independently of one another, from hydrogen, halogen, C 1-8 Alkyl or C 1-8 An alkyloxy group.
4. A compound of formula I, its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound according to any one of claims 1 to 3, wherein the structure of the compound of formula I is represented by formula I-1 or formula I-2:
Figure FDA0003608368330000052
of these, B, D, E, G, J, X, R 1 Having the structure of any one of claims 1 to 3In the definition, n is an integer of 1 to 4;
preferably, the structure of the compound of formula I is as shown in formula I-3, formula I-4, formula I-5, formula I-6, formula I-7 or formula I-8:
Figure FDA0003608368330000061
of these, E, G, J, X, R 1 Having the definition as set forth in any one of claims 1 to 3.
5. The compound of formula I, its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound according to any one of claims 1 to 4, wherein the structure of the compound of formula I is represented by formula II or formula III:
Figure FDA0003608368330000062
Wherein the content of the first and second substances,
E. g has the definition of any one of claims 1 to 4;
preferably, E is selected from O, C (O), NH, NCH 3 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 NHC (O), N (-cyclopropyl) CH 2 、NHCH 2 Or N (CH) 3 )CH 2 ；
G is selected from optionally substituted by one, two or more R 4 Substituted C 9-14 An aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl group, said aryl, heteroaryl or heterocyclyl group being a group of a fused ring;
R 4 having the definition of any one of claims 1 to 4;
preferably, E is selected from O, C (O), NH, NCH 3 、S(O) 2 、CH 2 、NHS(O) 2 、OCH 2 、O(CH 2 ) 2 、NHC(O)、NHCH 2 Or N (CH) 3 )CH 2 ；
G is selected from optionally substituted by one, two or more R 4 Substituted C 9-14 An aryl, 9-14 membered heteroaryl or 9-14 membered heterocyclyl, said aryl, heteroaryl or heterocyclyl being groups of fused rings;
R 4 having the definition as set forth in any one of claims 1 to 4.
6. The compound of formula I, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof according to any one of claims 1 to 5, wherein G is selected from the group consisting of:
Figure FDA0003608368330000071
Figure FDA0003608368330000081
the G may also be selected from the following groups:
Figure FDA0003608368330000082
Figure FDA0003608368330000083
7. the compound of formula I, its racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds according to any one of claims 1 to 6, wherein the structure of the compound of formula I is as follows:
Figure FDA0003608368330000084
Figure FDA0003608368330000091
Figure FDA0003608368330000101
Figure FDA0003608368330000111
Figure FDA0003608368330000121
Figure FDA0003608368330000131
Figure FDA0003608368330000141
8. A process for the preparation of a compound of formula I as claimed in any one of claims 1 to 7, comprising scheme 1 or scheme 2 below:
scheme 1: the compound 1 generates a ring reaction under the action of alkali to obtain a compound shown as a formula I-1;
Figure FDA0003608368330000151
wherein B, E, G, J, X, R 1 And n has the meaning as claimed in any of claims 1 to 7, R is C 1-8 An alkyl group;
preferably, the base in scheme 1 may be an inorganic base, preferably an alkali metal hydroxide, such as lithium hydroxide, lithium hydroxide monohydrate, sodium hydroxide, potassium hydroxide;
scheme 2: reacting the compound 2 with the compound 3 to obtain a compound shown as a formula I-2;
Figure FDA0003608368330000152
wherein D, E, G, J, X, R 1 And n has the definition as set forth in any one of claims 1 to 7;
y is a reactive group capable of attaching, by substitution or coupling, compound 3 to the D ring of compound 2 in a reaction, e.g. halogen, NH 2 、B(OH) 2 ；
Preferably, the reaction in scheme 2 may be carried out in the presence of a base, such as an organic base, which may be at least one of triethylamine, diisopropylethylamine, pyridine, DMAP, DBU; an organometallic compound such as at least one of lithium diisopropylamide, methyllithium, butyllithium;
preferably, the reaction described in scheme 1 or scheme 2 may be carried out in the presence of a solvent such as an organic solvent or a mixed solvent of an organic solvent and water.
9. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound of formula I as described in any one of claims 1-7, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof.
10. At least one compound of formula I according to any one of claims 1 to 7, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof, and the use of a pharmaceutical composition according to claim 9 for the manufacture of a medicament;
preferably, the use may be in the manufacture of a medicament for the treatment of a sex-hormone related disorder, such as in the manufacture of a GnRH receptor antagonist.
11. Use according to claim 10, for the preparation of a medicament for the treatment or prevention of sex hormone dependent cancer, bone metastasis of sex hormone dependent cancer, prostatic hypertrophy, uterine fibroids, endometriosis, uterine fibroids, precocious puberty, amenorrhea, premenstrual syndrome, dysmenorrhea, atrial ovarian syndrome, polycystic ovary syndrome, acne, alopecia, alzheimer's disease, infertility, irritable bowel syndrome, hormone-independent and LH-RH sensitive benign or malignant tumors, or diseases of flushing; such as the use in the preparation of reproductive regulators, contraceptives, ovulation inducers; or as a medicament for the preparation of a medicament for the prevention of postoperative recurrence of a sex hormone dependent cancer, wherein the sex hormone dependent cancer is selected from the group consisting of prostate cancer, uterine tumors, breast cancer, pituitary cancer.
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