CN115232144B - Nitrogen-containing condensed ring derivative, pharmaceutical composition, and preparation method and application thereof - Google Patents

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

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CN115232144B
CN115232144B CN202210425594.5A CN202210425594A CN115232144B CN 115232144 B CN115232144 B CN 115232144B CN 202210425594 A CN202210425594 A CN 202210425594A CN 115232144 B CN115232144 B CN 115232144B
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CN115232144A (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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • 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 in a formula I, a pharmaceutical composition, a preparation method and application thereof. The compound has good GnRH receptor antagonism, has longer half-life, and can be used for treating or preventingConditions and diseases associated with gonadotropins and the preparation of medicaments for such conditions and diseases.

Description

Nitrogen-containing condensed ring derivative, pharmaceutical composition, and preparation method and application thereof
The invention claims priority from the prior application filed in 2021, 4 and 22 to China national intellectual property agency, with patent application number 202110438605.9, named as 'nitrogen-containing condensed ring derivatives, pharmaceutical compositions, preparation methods and applications thereof'. The entirety of this prior application is incorporated by reference into the present invention.
Technical Field
The invention belongs to the field of medicines, and particularly relates to a nitrogen-containing condensed ring derivative, a pharmaceutical composition, a preparation method and application thereof.
Background
Gonadotropin releasing hormone (Gonadoliberin; 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 hypothalamic neuroendocrine cells 2 ) Is endocrine germ lineCentral regulatory factors in the system. It is delivered to the pituitary via the hypothalamic pituitary portal circulatory system, binds to GnRH receptor cells of the anterior pituitary, and the secretion and release of gonadotropins such as luteinizing hormone (Luteinizing Hormone, LH) and Follicle-stimulating hormone (FSH), which regulate the normal development of the ovary and corpus luteum, play an important role in the hypothalamic-pituitary-gonadal axis. GnRH receptors exert their regulatory effects by coupling to G proteins that activate the calcium phosphatidylinositol second messenger system, while LH regulates the production of sex steroids and FSH regulates the development of male spermatogenesis and female follicles.
LH and FSH are released into the circulation and bind to receptors on specific cells of the ovary or testis, stimulating the production of steroids. In the presence of sex steroids, conditions such as endometriosis, uterine fibroids and prostate cancer are exacerbated and need to be controlled therapeutically by administration of long-acting peptide GnRH receptor agonists and antagonists.
Peptide GnRH receptor antagonists include cetrorelix, ganirelix, etc., which block GnRH binding to its receptor by primarily rapidly competing for binding to the GnRH receptor, blocking dimer complex formation and signaling, and thus inhibiting release of LH and FSH. However, peptide compounds have a number of problems to be solved including oral absorbability, dosage form, dosage volume, drug stability, sustained action, and metabolic stability. The main reason why the small molecule GnRH receptor antagonist is superior to the existing peptide therapies is that the small molecule GnRH receptor antagonist can be directly and orally administered, which is convenient and quick. The 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 peptide GnRH receptor antagonists, some non-peptide GnRH receptor antagonists have been proposed and entered into the development, clinical trial and marketing stages, for example Elagolix developed by AbbVie corporation has been batched for the treatment of endometriosis, while uterine myoma indications have been declared on the market; the Relugolix developed by Takada corporation has been approved for the treatment of uterine fibroids and indications for endometriosis and prostate cancer have entered a three-phase clinical stage; linzagolix, developed by the company ObsEva and Kissei, was in a three-phase clinical trial phase for both endometriosis and uterine fibroids.
Although significant research has been conducted in this area, there is still a continuing need to develop more potent small molecule GnRH receptor antagonists, the present invention provides a novel structural compound as a GnRH receptor antagonist, and it has been found that the compounds having such structure have good activity and are effective in treating endocrine-reproductive system diseases.
Disclosure of Invention
In order to solve the above technical problems, the present invention provides a compound represented by formula I, its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof:
Wherein Y is selected from the following groups which are unsubstituted or substituted by X and/or-O-J: c (C) 6-20 An aromatic ring or a 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 with one, two or more R 1 SubstitutedOr->
B is selected from unsubstituted or optionally substituted with one, two or more R 2 Substituted 5-20 membered heteroaryl rings;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-20 membered heterocycle or 5-20 membered heteroaryl 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, C 6-20 Aryl, 5-20 membered heterocyclyl or 5-20 membered heteroaryl;
j is selected from H, unsubstituted or optionally substituted with one, two or more R 5 Substituted C 1-40 Alkyl, C 3-20 Cycloalkyl, C 6-20 Aryl, 5-20 membered heteroaryl, or J-O-is fused with the Y to which it is attached, preferably with the benzene ring to which it is attached, to form a 5-20 membered heteroaryl ring;
x is selected from hydrogen or halogen;
is a connecting site;
each R 1 、R 2 、R 3 、R 4 、R 5 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), unsubstituted or optionally substituted by one, two or more R a Substituted with the following groups: c (C) 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyl oxy, 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 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), unsubstituted or optionally substituted by one, two or more R b Substituted with the following groups: c (C) 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyl oxy, 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 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), COOH, COOC 1-40 Alkyl, C 1-40 Alkyl, C 2-40 Alkenyl, C 2-40 Alkynyl, C 3-40 Cycloalkyl, C 3-40 Cycloalkenyl, C 3-40 Cycloalkynyl radicals, C 6-20 Aryl, 5-20 membered heteroaryl, 3-20 membered heterocyclyl, C 1-40 Alkyloxy, C 2-40 Alkenyloxy, C 2-40 Alkynyloxy, C 3-40 Cycloalkyl oxy, C 3-40 Cycloalkenyl oxy, 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: benzene or pyridine rings;
b is selected from unsubstituted or optionally substituted with one, two or more R 2 Substituted 5-14 membered heteroaryl rings;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 A substituted 5-14 membered heterocycle or 5-14 membered heteroaryl 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, C 6-14 Aryl, 5-14 membered heterocyclyl or 5-14 membered heteroaryl; when A is selected fromAnd B is selected from thiophene ring, G is selected from 9-14 membered heteroaryl or 9-14 membered heterocyclyl, preferably fused ring groups;
j is selected from H, unsubstituted or optionally substituted with one, two or more R 5 Substituted C 1-8 Alkyl, C 3-8 Cycloalkyl, C 6-14 Aryl, 5-14 membered heteroaryl, or J-O-is linked to C in the ortho position in the benzene ring to which it is attached, forming a 5-14 membered heteroaryl ring in parallel with the benzene ring;
x is selected from hydrogen or halogen;
is a connecting site;
each R 1 、R 2 、R 3 、R 4 、R 5 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), unsubstituted or optionally substituted by one, two or more R a Substituted with the following groups: c (C) 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkenyl, C 3-8 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyl oxy, C 3-8 Cycloalkynyloxy, C 6-14 Aryloxy groupRadical, 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 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), unsubstituted or optionally substituted by one, two or more R b Substituted with the following groups: c (C) 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkenyl, C 3-8 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyl oxy, 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 The same or different, independently of one another, are selected from hydrogen, halogen, OH, CN, NO 2 Oxo (=o), COOH, COOC 1-8 Alkyl, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkenyl, C 3-8 Cycloalkynyl radicals, C 6-14 Aryl, 5-14 membered heteroaryl, 3-8 membered heterocyclyl, C 1-8 Alkyloxy, C 2-8 Alkenyloxy, C 2-8 Alkynyloxy, C 3-8 Cycloalkyl oxy, C 3-8 Cycloalkenyl oxy, 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 the group consisting of unsubstituted or optionally substituted with one, two or more R 2 A substituted thiophene ring, furan ring, imidazole ring, pyrazole ring;
d is selected from unsubstituted or optionally substituted by one, two or more R 3 Substituted thiophene ring, furan ring, imidazole ring, pyrazole ring, pyrrole ring, tetrahydropyrrole ring;
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 fromAnd B is selected from thiophene rings, G is selected from unsubstituted or optionally substituted by 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 be a fused ring group;
j is selected from H, unsubstituted or optionally substituted with one, two or more R 5 Substituted C 1-8 Alkyl, C 3-8 Cycloalkyl or J-O-is linked to C ortho to 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 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;
is a connecting site;
each R 1 The same or different, independently of one another, from hydrogen, C 1-8 Alkyl group 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 group、Ph-NH-C(O)-NH-OC 1-8 Alkyl, 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 (b)
Each R 2 The same or different, independently of one another, from hydrogen, C 1-8 Alkyl group 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 group, ph-NH-C (O) -NH-OC 1-8 Alkyl, 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 (b)
Each R 3 The same or different, independently of one another, 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 with the following groups: c (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 alkyl oxy 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 with the following groups: c (C) 1-8 Alkyl, 5-14 membered heteroaryl, C 1-8 An alkyl oxy group;
each R a The same or different, independently of one another, from hydrogen, halogen, C 1-8 Alkyl, C 3-8 Cycloalkyl, C 1-8 An alkyl oxy group.
According to an embodiment of the invention, the structure of the compound of formula I is as shown in formula I':
wherein A, E, G, J, X has the definition set forth above.
According to an embodiment of the present invention, the structure of the compound of formula I is as shown in formula I-1 or I-2:
therein, B, D, E, G, J, X, R 1 With the definition described above, n is selected from integers from 1 to 4.
According to an embodiment of the present invention, 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:
therein, E, G, J, X, R 1 With the definition described above.
According to an embodiment of the invention, the structure of the compound of formula I is as shown in formula II:
wherein,
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 above-described definition, preferably being unsubstituted or optionally substituted by 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 being a fused ring group;
R 4 with the definition described above.
According to an embodiment of the invention, the structure of the compound of formula I is as shown in formula II:
wherein G has the definition as set forth above, preferably 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 being a fused ring group;
R 4 with the definition described above.
In a preferred embodiment of the invention, said G is selected from the group consisting of:
according to an embodiment of the invention, the structure of the compounds of formula I is as follows:
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the present invention also provides a process for the preparation of a compound 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;
therein, B, E, G, J, X, R 1 And n has the above-described definition, 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 a compound 3 to obtain a compound shown in a formula I-2;
therein, D, E, G, J, X, R 1 And n has the definition as described above; y is a reactive group, which in the reaction enables 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 performed in the presence of a base, for example, 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, methyl lithium, butyl lithium.
According to an embodiment of the present invention, the reaction of the scheme 1 or the scheme 2 may be performed 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, and n-butanol; ethers such as ethyl propyl ether, n-butyl ether, anisole, phenetole, cyclohexylmethyl ether, dimethyl ether, diethyl ether, dimethylethylene glycol, diphenyl ether, propyl ether, isopropyl ether, isobutyl ether, isopentyl ether, ethylene glycol dimethyl ether, isopropyl ethyl ether, methyl tert-butyl ether, tetrahydrofuran, methyltetrahydrofuran, 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 methylene chloride, chloroform, carbon tetrachloride, fluorobenzene, chlorobenzene or dichlorobenzene; cyclohexane, methylcyclohexane, petroleum ether, acetone, octane, benzene, toluene, chlorobenzene, bromobenzene, xylene; esters such as methyl acetate, ethyl acetate, butyl acetate, isobutyl acetate, and dimethyl carbonate, dibutyl carbonate, or vinyl carbonate.
The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of a compound of 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 embodiments of the present invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.
The present invention also provides a method of treating a gonadotropin-associated disorder comprising administering to a subject a prophylactically or therapeutically effective amount of at least one of a compound of formula (I), racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.
The invention also provides a method of treating a gonadotropin-associated disorder comprising administering to a subject a prophylactically or therapeutically effective amount of the above-described pharmaceutical composition.
Such gonadal hormone-related disorders include sex hormone dependent cancer, bone metastases of sex hormone dependent cancer, prostatic hypertrophy, uterine fibroids, 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 the group consisting of prostate cancer, uterine tumor, breast cancer, and pituitary cancer.
In some embodiments, the patient comprises a mammal, preferably a human.
The present invention also provides at least one of a compound represented by formula (I), a racemate, a stereoisomer, a tautomer, an isotopic label, a solvate, a polymorph, a pharmaceutically acceptable salt or a prodrug compound thereof, or a pharmaceutical composition thereof, for preventing or treating a gonadotropin-associated disease.
The invention also provides application of at least one of the compounds shown in the formula (I), racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof in preparing medicines.
According to an embodiment of the present invention, the use may be in the manufacture of a medicament for the prevention or treatment of a gonadotropin associated disorder, such as in the manufacture of a medicament for 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 fibroids, 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 diseases; such as in the preparation of reproductive modulators, contraceptives, ovulation inducers; or as a medicament for the prevention of postoperative recurrence of sex hormone dependent cancers selected from the group consisting of prostate cancer, uterine tumor, breast cancer, pituitary cancer.
The medicament may be used in diseases associated with gonadotropins.
The effective dosage of the active compound can range widely, and is generally administered in a pharmaceutically effective amount. However, it will be appreciated that the amount of the compound actually administered will generally be determined by the physician, in light of the relevant circumstances, and will include the condition to be treated, the route of administration selected, the actual compound administered; age, weight, and response of the individual patient; severity of patient symptoms, and the like.
The amount of the compound or composition administered to the patient is not fixed and depends on the drug administered, the purpose of the administration, e.g., prophylaxis or treatment; the condition of the patient, the mode of administration, etc. In therapeutic applications, the compositions may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially inhibit 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, depending on factors such as the severity of the disease, the age, weight and general condition of the patient.
The composition to be 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 filter sterilized.
Therapeutic doses of the compounds of the invention may be determined, for example, according to the following: the specific use of the treatment, the manner in which the compound is administered, the health and condition of the patient, and the discretion of the prescribing physician. The proportion or concentration of the compounds of the invention in the pharmaceutical composition may be variable, depending on a number of factors, including the dosage, chemical characteristics (e.g. hydrophobicity) and route of administration. The compounds of the invention may be provided, for example, by a physiologically buffered aqueous solution containing about 0.1 to 10% w/v of the compound for parenteral administration. Some typical dosages range from about 1 μg/kg to about 1g/kg body weight/day. In certain embodiments, the dosage ranges from about 0.01mg/kg to about 100mg/kg body weight/day. Dosages will likely depend on such variables as the type and extent of progression of the disease or disorder, the general health of the particular patient, the relative biological efficacy of the compound selected, the excipient formulation and its route of administration. The effective dose can be obtained by extrapolation of the dose-response curve derived from in vitro or animal model test systems.
Advantageous effects
The compound provided by the invention has good GnRH receptor antagonism, can be used for treating or preventing diseases and symptoms related to gonadotropins, and can be used for preparing medicines for treating the diseases and the symptoms. And, the half-life of the compound is longer.
Definition and description of terms
Unless otherwise indicated, the radical and term definitions recited in the specification and claims of this application, including as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and combinations of radical definitions and compound structures should be understood to be within the scope of the description and/or claims herein.
The numerical ranges recited in the specification and claims are equivalent to at least each specific integer number recited therein unless otherwise stated. For example, the numerical range "1 to 40" corresponds to the numerical range in which each of the integer numbers 1 to 10, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and each of the integer numbers 11 to 40, i.e., 11, 12, 13, 14, 15, &..times., 35, 36, 37, 38, 39, 40 are described. Furthermore, when certain numerical ranges are defined as "numbers," it is to be understood that both endpoints of the range, each integer within the range, and each fraction within the range are delineated. For example, a "number of 0 to 10" should be understood to describe not only each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also 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 the description of 1,2 or more herein, "more" shall mean an integer greater than 2, such as greater than or equal to 3, such as 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 that it may be unsubstituted (unsubstituted) or optionally substituted with one, two or more R.
The term "halogen" means fluorine, chlorine, bromine and iodine.
The term "C 1-40 Alkyl "is understood to mean a straight-chain or branched saturated monovalent hydrocarbon radical having from 1 to 40 carbon atoms. For example, "C 1-10 Alkyl "means straight-chain and branched alkyl having 1,2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms," C 1-8 Alkyl "means straight and branched alkyl having 1,2, 3, 4, 5, 6, 7, or 8 carbon atoms," C 1-6 Alkyl "means straight and branched alkyl groups having 1,2, 3, 4, 5 or 6 carbon atoms. The alkyl 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-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 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 mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has from 2 to 40 carbon atoms, preferably" C 2-10 Alkenyl groups). "C 2-10 Alkenyl "is understood to mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2,3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably" C 2-8 Alkenyl groups). "C 2-10 Alkenyl "is understood to mean preferably a straight-chain or branched monovalent hydrocarbon radical which contains one or more double bonds and has 2,3, 4, 5, 6, 7 or 8 carbon atoms, for example 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 will be appreciated that where the alkenyl group comprises more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl 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, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3-enyl, (E) -hex-2-enyl, (Z) -hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylpropan-1-enyl, 3-methylbutan-3-enyl, 2-methylbutan-3-enyl, 1-methylbutan-3-enyl, 3-methylbutan-2-enyl, (E) -2-methylbutan-2-enyl, (Z) -2-methylbutan-2-enyl, (E) -1-methylbutan-2-enyl, (Z) -1-methylbutan-2-enyl, (E) -3-methylbutan-1-enyl, (Z) -3-methylbutan-1-enyl, (E) -2-methylbutan-1-enyl, (Z) -2-methylbutan-1-enyl (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.
The term "C 2-40 Alkynyl "is understood to mean a monovalent hydrocarbon radical, directly or branched, containing one or more triple bonds and having from 2 to 40 carbon atoms, preferably" C 2-10 Alkynyl groups. The term "C 2-10 Alkynyl "is understood to mean preferably a straight-or branched-chain 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-methylpropan-2-ynyl, 2-methylbutan-3-ynyl, 1-methylbutan-2-ynyl, 3-methylbutan-1-ynyl, 1-ethylpropan-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, 1-isopropylprop-2-ynyl, 2-dimethylbbut-3-ynyl, 1, 1-dimethylbut-3-ynyl, 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 ring or tricyclic hydrocarbon ring having 3 to 40 carbon atoms, preferably" C 3-10 Cycloalkyl ", 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 ring,Spiro) hydrocarbon rings or tricycloalkanes having 3,4, 5, 6, 7, 8, 9 or 10 carbon atoms. The C is 3-10 Cycloalkyl can be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as campholyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo [2.1.1 ]]Hexyl, bicyclo [2.2.1]Heptyl, bicyclo [2.2.1]Heptenyl, 6-dimethylbicyclo [3.1.1]Heptyl, 2, 6-trimethylbicyclo [3.1.1]Heptyl, bicyclo [2.2.2]Octyl, 2, 7-diazaspiro [3,5 ]]Nonylalkyl, 2, 6-diazaspiro [3,4 ]]Octyl, or tricyclic hydrocarbon groups such as adamantyl.
The term "3-20 membered heterocyclyl" refers to a saturated or unsaturated, non-aromatic ring or ring system, unless otherwise defined, which is, for example, 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 which contains at least one, for example, 1, 2, 3,4, 5, or more heteroatoms selected from O, S and N, wherein N and S may also optionally be oxidized to various oxidation states to form nitrogen oxides, -S (O) -or-S (O) 2 -a state of the device. Preferably, the heterocyclic group may be selected from "3-10 membered heterocyclic groups". 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 remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present. The heterocyclic group may include fused or bridged rings as well as spiro rings. In particular, the heterocyclic groups may include, but are not limited to: 4-membered rings such as azetidinyl, oxetanyl; a 5-membered ring 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 heterocyclyl may be benzo-fused. The heterocyclic group may be bicyclic, such as, but not limited to, a 5,5 membered ring, such as hexahydroCyclopenta [ c ]]Pyrrol-2 (1H) -yl ring, or 5,6 membered bicyclic ring, e.g. hexahydropyrrolo [1,2-a ]]Pyrazin-2 (1H) -yl ring. The heterocyclic 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 may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. When the 3-20 membered heterocyclic group is linked to other groups to form the compound of the present invention, the carbon atom on the 3-20 membered heterocyclic group may be linked to other groups, or the heterocyclic atom on the 3-20 membered heterocyclic ring may be linked to other groups. For example, when the 3-20 membered heterocyclic group is selected from piperazinyl, it may be that the nitrogen atom on the piperazinyl group is attached to other groups. Or when the 3-20 membered heterocyclic group is selected from piperidyl, it may be that the nitrogen atom on the piperidyl ring and the carbon atom at the para position thereof are attached to other groups.
The term "C 6-20 Aryl "is understood to mean preferably a mono-, bi-, e.g. fused-, bridged-, spiro-or tricyclic hydrocarbon ring of monovalent aromatic or partly aromatic character having 6 to 20 carbon atoms, which may be a monoaromatic ring or a polyaromatic ring fused together, preferably" C 6-14 Aryl group). The term "C 6-14 Aryl "is understood to mean preferably a mono-, bi-or tricyclic hydrocarbon ring (" C ") having a monovalent aromatic or partially aromatic character of 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms 6-14 Aryl), in particular a ring having 6 carbon atoms ("C) 6 Aryl "), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C 9 Aryl "), e.g. indanyl or indenyl, or a ring having 10 carbon atoms (" C 10 Aryl "), such as tetralin, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms (" C " 13 Aryl "), e.g. fluorenyl, or a ring having 14 carbon atoms (" C) 14 Aryl "), such as anthracenyl. When said C 6-20 When aryl is substituted, it may be mono-substituted or poly-substituted. The substitution site 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, such as "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: it has 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 it contains 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, in addition, can be benzo-fused in each case. "heteroaryl" also refers to groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, wherein the attached radical or point 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-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7-, or 8-phthalazinyl (phtalazinyl), 2-, 3-, 4-, 5-, or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-, or 8-naphthyridinyl, 2-, 4-, 6-, 7-, or 7-, 1-, 3-, 4-, 3-, 5-, 6-, 1-and 2-amino 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-pyrimidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-or 10-phenanthrolinyl, 1-, 2-, 3-, 4-, 5-, 8-, 9-or 10-phenanthrolinyl 2-, 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 [2,3-b ] furanyl, 2-, 3-, 5-, 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 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-benzoxepin (benzoxapinyl), 2-, 4-, 5-, 6-, 7-or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-or 11-1H-pyrrolo [1,2-b ] [2] benzazapinyl. 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 is attached to other groups to form the compounds of the invention, the carbon atom on the 5-20 membered heteroaryl ring may be attached to other groups, or the heteroatom on the 5-20 membered heteroaryl ring may be attached to other groups. When the 5-20 membered heteroaryl is substituted, it may be mono-substituted or poly-substituted. And, the substitution site thereof is not limited, 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-forming 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 members.
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 that may include substitution at 1, 2, or more of its 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) or bonding to other groups include pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thienyl or thienylene include thiophen-2-yl, thienylene-2-yl, thiophen-3-yl and thienylene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.
The term "oxo" refers to the substitution of a carbon atom, nitrogen atom or sulfur atom in a substituent with an oxo group (=o) formed after oxidation.
"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 "refers to 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 (Pd) 2 (dba) 3 "means tris (dibenzylideneacetone) dipalladium. "Dppf" refers to 1,1' -bis-diphenylphosphino ferrocene. "HATU" refers to 2- (7-oxo-benzotriazol) -N, N' -tetramethylurea hexafluorophosphate. "KHMDS" refers to potassium hexamethyldisilazide. "LiHMDS" refers to lithium bis (trimethylsilylamide). "MeLi" refers to lithium-based. "n-BuLi" refers to n-butyllithium. "NaBH (OAc) 3 "means sodium triacetoxyborohydride.
Unless otherwise indicated, the definitions of terms herein apply equally to the groups containing 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.
Those skilled in the art will appreciate that the compounds of formula (I) may exist in various pharmaceutically acceptable salt forms. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; these compounds may also form internal salts if they contain both acidic (e.g., carboxyl) and basic (e.g., amino) centers.
The compounds of the invention may exist in the form of solvates (e.g. hydrates) wherein the compounds of the invention comprise a polar solvent as a structural element of the compound lattice, in particular, for example, water, methanol or ethanol. The polar solvent, in particular water, may be present in stoichiometric or non-stoichiometric amounts.
Depending on its molecular structure, the compound of the invention may be chiral and thus various enantiomeric forms may exist. These compounds may thus be present in racemic or optically active form. The compounds of the present invention encompass isomers or mixtures, racemates thereof wherein each chiral carbon is in the R or S configuration. The compounds of the invention or intermediates thereof may be isolated as enantiomer 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 the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as 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 camphorsulfonic acids in R and S form. The chromatographic 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 chiral derivatized methacrylate polymers, immobilized on silica. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.
The corresponding stable isomer may 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" refers to 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 disease: 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 a pathology or symptomatology of the disease. (2) inhibition of disease: for example, inhibiting a disease, disorder or condition (i.e., preventing further development of pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition. (3) alleviation of disease: for example, alleviating a disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition.
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention.
Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.
The structure of the compounds of the present invention is determined by Nuclear Magnetic Resonance (NMR) or/and liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR was performed using Bruker AVANCE-400 nuclear magnetic resonance apparatus with deuterated dimethyl sulfoxide (DMSO-d) 6 ) Deuterated methanol (CD) 3 OD) and deuterated chloroform (CDCl) 3 ) The internal standard is Tetramethylsilane (TMS).
An Agilent 1200 affinity Series mass spectrometer was used for LC-MS measurement. HPLC was performed using Agilent 1200DAD high pressure liquid chromatography (Sunfire C18X 4.6mm column) and Waters 2695-2996 high pressure liquid chromatography (Gimini C18X 4.6mm column).
The thin layer chromatography silica gel plate uses a smoke table 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. Column chromatography uses yellow sea silica gel of 200-300 meshes as carrier.
All reactions of the present invention were carried out under continuous magnetic stirring under a dry nitrogen or argon atmosphere, the solvent being a dried solvent, and the reaction temperature being in degrees celsius, without specific description.
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)
First step
Preparation of 6- (bromomethyl) quinoline
Methylol quinoline 029a (500 mg,3.14 mmol) and phosphine tribromide (1.28 g,47 mmol) were dissolved in 1, 4-dioxane (20 mL). The reaction mixture was reacted at 25℃for 0.5h. After the reaction was completed, the pH was adjusted to 7-8 with saturated sodium bicarbonate solution, followed by extraction with methylene chloride (2X 20 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated to give 6- (bromomethyl) quinoline 029b (615 mg, yellow solid), yield: 79%.
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
Preparation of 6- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline
4-fluoro-2-methoxy-5-nitrophenol 029c (300 mg,1.6 mmol) and 6- (bromomethyl) quinoline 029b (399mg, 1.76 mmol) were dissolved in acetonitrile (20 mL), and potassium carbonate (428 mg,3.2 mmol) 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 and dried over anhydrous sodium sulfate, concentrated to give the crude product which was purified by 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).
Third step
Preparation of 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) aniline
6- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 029d (200 mg,6.1 mmol) and iron powder (85.1636 mg,1.525 mmol) were dissolved in acetonitrile (20 mL) and saturated ammonium chloride solution (5 mL) was added thereto. The reaction mixture was reacted at 25℃for 16 hours. The reaction solution was filtered to remove iron powder, and then extracted with methylene chloride (2X 60 mL). The organic phases were combined and 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% yield.
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).
Fourth step
Preparation of dimethyl 4- ({ [ 2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-4-methoxy-5- (quinolin-6-ylmethoxy) aniline 029e (20 mg,0.07 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylic acid (25.82 mg,0.077 mmol) were dissolved in tetrahydrofuran (2 mL), and triethylamine (21.21 mg,0.21 mmol) was added thereto and the reaction solution was reacted at 25℃for 24 hours. This step was used directly in the next reaction without post-treatment.
MS m/z(ESI)：540.12[M+1] + 。
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
To the reaction solution of the previous step were added methanol (2 mL) and water (2 mL), followed by addition of lithium hydroxide monohydrate (16.99 mg,0.405 mmol). The reaction mixture was reacted at 25℃for 1 hour. After the reaction, the pH was adjusted to 6-7 with 1N hydrochloric acid, and extracted with methylene chloride (2X 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by prep. (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, pale 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-002)
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First step
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxybenzene
4-fluoro-2-methoxyphenol 30a (2 g,14 mmol) was added to acetone (20 mL), and benzyl bromide (2.9 g,17 mmol) and potassium carbonate (2.92 g,21.11 mmol) were then added, and the reaction stirred at 65℃for 16 hours. After completion of the reaction, water (50 mL) was added to the reaction solution, extraction was performed 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 give 1- (benzyloxy) -4-fluoro-2-methoxybenzene 30b (3 g, yellow solid), yield: 83%.
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
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 ℃ and nitric acid (0.2 mL, 65%) was added, and after completion of the reaction, water (10 mL) was added to the reaction solution, which was 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), yield: 83%.
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).
Third step
Preparation of 4-fluoro-2-methoxy-5-nitrophenol
1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene 30c (550 mg,1.98 mmol) was dissolved in boron trichloride (3 mL,1M in DCM) and the reaction stirred at 25℃for 2 h. LCMS monitored completion of reaction water (10 mL) was added to the reaction, 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 give 4-fluoro-2-methoxy-5-nitrophenol 30d (200 mg, yellow solid), yield: 51%.
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).
Fourth step
Preparation of isoquinolin-8-yl methanol
Isoquinolin-8-yl-formaldehyde 30e (250 mg,1.59 mmol) was dissolved in methanol (10 mL), sodium borohydride (30 mg,0.80 mmol) 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 mixture, 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-ylmethanol 30f (210 mg, yellow oil), yield: 75%.
MS m/z(ESI)：160.2[M+1] + 。
Fifth step
Preparation of 8- (chloromethyl) isoquinoline
Isoquinolin-8-yl methanol 30f (210 mg,1.38 mmol) was dissolved in dichloromethane (20 mL), tsCl (399mg, 2.07 mmol), DMAP (50 mg,0.41 mmol) and triethylamine (319 mg,4.14 mmol) 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 mixture, which was extracted with methylene chloride (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%.
MS m/z(ESI)：178.2[M+1] + 。
Sixth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
8- (chloromethyl) isoquinoline 30g (200 mg,1.13 mmol) was dissolved in acetonitrile (20 mL), 4-fluoro-2-methoxy-5-nitrophenol 30d (232 mg,1.24 mmol), potassium carbonate (312 mg,2.26 mmol) and potassium iodide (19 mg,0.11 mmol) 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 mixture, extracted with dichloromethane (3×20 mL), and 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 give 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline for 30h (150 mg, pale yellow solid), yield: 36%.
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 (130 mg,0.37 mmol) was dissolved in acetonitrile (10 mL), iron powder (62 mg,1.11 mmol) and saturated ammonium chloride solution (5 mL) were added and the reaction stirred at 25℃for 3 hours. After completion of LCMS monitoring the reaction, the solid in the reaction mixture was filtered off, extracted with dichloromethane (3×20 mL), 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, light yellow solid 81%), yield: 81%.
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 (100 mg,0.34 mmol) was dissolved in tetrahydrofuran (20 mL), and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (228 mg,0.68 mmol) and triethylamine (103 mg,1.02 mmol) were added, and the reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, the reaction mixture was concentrated to give 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: 59%.
MS m/z(ESI)：540.1[M+1] + 。
Ninth step
Preparation of 3- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -2, 4-dioxo-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 (120 mg,0.222 mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL) and methanol (5 mL), lithium hydroxide (24 mg,0.56 mmol) was added, and the reaction solution was stirred at 25℃for 3 hours. After completion of LCMS monitoring the reaction, the reaction mixture was adjusted to pH 5-6 with 1N HCl, extracted with ethyl acetate (3×20 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-002 (30 mg, pale yellow solid), yield: 32%.
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.
Example 3 and example 4
Preparation of 3- (5- { [ (4S) -5, 6-difluoro-3, 4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-004)
First step
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 (2.1 g,52.5 mmol) (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, then extracted with ethyl acetate (3×50 mL). The combined organic phases were taken up 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%.
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
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 (3.6 g,13.5 mmol) in dichloromethane (100 mL) was added a catalytic amount of DMF. Oxalyl chloride (3.4 g,27 mmol) was slowly added dropwise to the reaction solution at 0 ℃. After completion of the dropwise addition, the reaction mixture was stirred at 25℃for 1h. Aluminum trichloride (2 g,14.8 mmol) was added at 0deg.C. 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 the organic phases were combined, washed with 0.5M aqueous NaOH (2X 50 mL), brine (2X 40 mL), dried over anhydrous sodium sulfate and concentrated. The residue obtained was separated and purified by silica gel column (petroleum ether/ethyl acetate=6/1) to give 8-bromo-5, 6-difluoro-2, 3-dihydro-1-benzopyran-4-one 031c (3.1 g, yellow solid), yield: 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).
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 (0.5 g,1.9 mmol) was dissolved in MeOH (5 mL), to which sodium borohydride (108 mg,2.8 mmol) was slowly added. 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 organics 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%.
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).
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 (450 mg,1.7 mmol) in THF (5 mL) was added 4-fluoro-2-methoxy-5-nitrophenol (318 mg,1.7 mmol) and triphenylphosphine (580 mg,2.21 mmol). DIAD (447 mg,2.21 mmol) was slowly added at 0deg.C. The reaction solution was then stirred at 45℃for 1h. The reaction mixture was concentrated. The resulting residue was separated and purified by 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%.
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).
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 (690 mg,1.59 mmol) 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 concentrated by filtration. The resulting residue was separated and purified by 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%.
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).
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 (100 mg,0.31 mmol) in THF (2 mL) was added dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (104 mg,0.31 mmol) and triethylamine (94 mg,0.93 mmol). 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 031g 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 (180 mg, yellow solid). Yield: 87%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution 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-dicarboxylic acid dimethyl ester 031g (24 mg,0.04 mmol) in tetrahydrofuran/methanol/water=1:1:2 (6 mL) was added lithium hydroxide (4 mg,0.16 mmol). 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 purified by chiral preparative HPLC (column: chiralpak-OD, mobile phase: carbon dioxide-methanol (0.1% DEA)) and then lyophilized and then conventional preparative HPLC (column: xbridge-C18X 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-003 (4 mg, off-white solid) and 3- (5- { [ (4R) -5, 6-difluoro-3, 4-dihydro-2H-1-benzopyran-4-yl ] oxy } -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-003 (4 mg, off-white solid) (004 mg, off-white solid): 19%.
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)
First step
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 (1.44 g,8.4 mmol) and potassium carbonate (2.91 g,21 mmol) were added. The reaction solution was stirred at 80℃for 6 hours. The reaction was quenched with water (100 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 column on silica gel (petroleum ether/ethyl acetate=4/1) to give the title product 2- (benzyloxy) -4-fluoro-1-methoxybenzene 035b (1.5 g, white solid) in 87% yield.
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
Preparation of 1- (benzyloxy) -5-fluoro-2-methoxy-4-nitrobenzene
2- (benzyloxy) -4-fluoro-1-methoxybenzene 035b (100 mg,0.43 mmol) was dissolved in acetic anhydride (3 mL), 65% nitric acid (109 mg,1.72 mmol) was added dropwise under ice water bath. The reaction solution was stirred at 25℃for 12 hours. The pH of the reaction solution was adjusted to 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) in 83% yield.
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).
Third step
Preparation of 5-fluoro-2-methoxy-4-nitrophenol
1- (benzyloxy) -5-fluoro-2-methoxy-4-nitrobenzene 035c (1 g,3.6 mmol) was dissolved in 1M boron trichloride in dichloromethane (10.8 mL,10.8 mmol). The reaction solution was stirred at 25℃for 12 hours. The reaction mixture was quenched with methanol (10 mL) and the concentrated residue was purified by column on silica gel (petroleum ether/ethyl acetate=3/2) to give the title product 5-fluoro-2-methoxy-4-nitrophenol 035d (0.6 g, yellow solid) in 80% yield.
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).
Fourth step
Preparation of 2, 3-difluoro-6-methoxybenzaldehyde
A solution of 2M lithium diisopropylamide (3.82 mL,7.64 mmol) in tetrahydrofuran (10 mL) was added dropwise to a solution of 1, 2-difluoro-4-methoxybenzene 035e (1000 mg,6.94 mmol) in tetrahydrofuran (3 mL) at-78℃and, after stirring at-78℃for 1 hour, N-dimethylformamide (578 mg,7.64 mmol) was added dropwise and stirring was continued for 1 hour. The reaction was quenched with acetic acid (1.5 mL) and water (50 mL), 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).
Fifth step
Preparation of 2-vinyl-3, 4-difluoro-1-methoxybenzene
Tris (triphenylphosphine) methyl bromide (1.71 g,4.80 mmol) was dissolved in tetrahydrofuran (15 mL) and cooled to-15℃and a 1M solution of potassium tert-butoxide (6.0 mL,6.00mmol,3 eq) in tetrahydrofuran was added dropwise. The reaction solution was stirred at 25℃for 1 hour. A solution of 2, 3-difluoro-6-methoxybenzaldehyde 035f (689 mg,4.00 mmol) in tetrahydrofuran (3 mL) was added thereto after cooling to-15℃and the reaction mixture was stirred at 25℃for another 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (100 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 column chromatography on silica gel (petroleum ether/ethyl acetate=50/1) to give the title product 2-vinyl-3, 4-difluoro-1-methoxybenzene 035g (4.25 g, pale 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).
Sixth step
Preparation of 2- (2, 3-difluoro-6-methoxyphenyl) ethanol
2-vinyl-3, 4-difluoro-1-methoxybenzene 035g (1000 mg,5.88 mmol) was dissolved in tetrahydrofuran (5 mL), a solution of 0.5M 9-BBN (35 mL,17.64 mmol) in tetrahydrofuran was added dropwise, the reaction was stirred at 25℃for 16 hours, then 3N NaOH (9.8 mL) and H were added 2 O 2 (3332 mg,29.4 mmol). The reaction was stirred at 25℃for a further 1 hour. The reaction mixture was quenched with water (100 mL), adjusted to pH 6, 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=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
Preparation of 1, 2-difluoro-3- [2- (5-fluoro-2-methoxy-4-nitrophenoxy) ethyl ] -4-methoxybenzene
2- (2, 3-difluoro-6-methoxyphenyl) ethanol 035h (250 mg,1.34 mmol) was dissolved in tetrahydrofuran (5 mL), 5-fluoro-2-methoxy-4-nitrophenol 035d (303 mg,1.61 mmol), triphenylphosphine (703 mg,2.68 mmol) and diisopropyl azodicarboxylate (552 mg,2.68 mmol) were added sequentially. The reaction solution was stirred in a microwave reactor at 60℃for 1 hour. The reaction solution was concentrated, and the obtained 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, pale yellow solid) in a yield of 41%.
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 (200 mg,0.56 mmol) was dissolved in methanol (10 mL) and wet palladium on carbon (60 mg,0.56 mmol) was added. The reaction solution was stirred under a hydrogen atmosphere at 25℃for 6 hours. 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, pale 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).
Ninth step
Preparation of dimethyl 4- [ ({ 4- [2- (2, 3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
4- [2- (2, 3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyaniline 035j (130 mg,0.40 mmol) was dissolved in tetrahydrofuran (5 mL), dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (135 mg,0.4 mmol) and triethylamine (122 mg,1.2 mmol) were added sequentially, and the reaction was stirred at 25℃for 16 hours.
MS m/z(ESI)：569.1[M+1] + .
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
Dimethyl 4- [ ({ 4- [2- (2, 3-difluoro-6-methoxyphenyl) ethoxy ] -2-fluoro-5-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 035k (200 mg,0.35 mmol) was dissolved in tetrahydrofuran (4 mL), lithium hydroxide monohydrate (74 mg,1.75 mmol) 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 extracted with ethyl acetate (3X 50 mL), the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the concentrated residue was purified with 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 } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-carboxylic acid (89 mg, 48 mg) as a yellow solid.
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)
First step
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 (2.4 g,7.3 mmol) was dissolved in acetonitrile (20 mL). Iron powder (1.22 g,21.9 mmol) and saturated ammonium chloride solution were then added. The reaction solution was reacted at 25℃for 8 hours. Then extracted with dichloromethane (2X 20 mL). The organic phases were combined and 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
Preparation of 2-amino-5-bromothiophene-3, 4-dicarboxylic acid diethyl ester
Diethyl 2-aminothiophene-3, 4-dicarboxylic acid 047c (330 mg,1.36 mmol) was dissolved in dichloromethane (5 mL), and N-bromosuccinimide (265.57 mg,1.49 mmol) was added at 0deg.C. The reaction mixture was reacted at 0℃for 1 hour. Then extracted with dichloromethane (2X 5 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure to give diethyl 2-amino-5-bromothiophene-3, 4-dicarboxylate 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).
Third step
Preparation of 2-bromo-5- [ (phenoxycarbonyl) amino ] -3, 4-dicarboxylic acid diethyl ester
Diethyl 2-amino-5-bromothiophene-3, 4-dicarboxylate 047d (400 mg,1.24 mmol) and phenyl chloroformate (582.4 mg,3.72 mmol) were dissolved in toluene (5 mL). The reaction mixture was reacted at 110℃for 3 hours. After the completion of the reaction, the mixture 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 diethyl 2-bromo-5- [ (phenoxycarbonyl) amino ] -3, 4-diformate 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).
Fourth step
Preparation of diethyl 2-bromo-5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-3, 4-dicarboxylate
Diethyl 2-bromo-5- [ (phenoxycarbonyl) amino ] thiophene-3, 4-dicarboxylate 047e (3838 mg,0.878 mmol) and 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline (250 mg,0.798 mmol) were dissolved in tetrahydrofuran (10 mL), and triethylamine (282.09 mg,2.79 mmol) was added thereto, and the reaction solution was reacted at 25℃for 24 hours. The crude product was obtained after concentration and passed through a silica gel column (petroleum ether/ethyl acetate=7/3) to give diethyl 2-bromo-5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-3, 4-dicarboxylic acid 047f (380 mg, yellow solid) in a yield of 64.79%.
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).
Fifth step
Preparation of ethyl 6-bromo-3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
2-bromo-5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy)]-2-fluoro-4-methoxyphenyl } carbamoyl) amino group]Thiophene-3, 4-dicarboxylic acid diethyl ester 047f (180 mg,0.272 mmol) and sodium methoxide (44.08 mg,0.82 mmol) were dissolved in methanol (2 mL). The reaction mixture was reacted at 40℃for 16 hours. After the reaction, the pH was adjusted to 6-7 with 1M diluted hydrochloric acid, quenched, and extracted with methylene chloride (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified over a silica gel column (petroleum ether/ethyl acetate=1/1) to give 6-bromo-3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-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 ]] + 。
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
Ethyl 6-bromo-3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 047g (110 mg,0.178 mmol), (6-aminopyridin-3-yl) borondiol (49.32 mg, 0.356 mmol), potassium carbonate (74.14 mg,0.54 mmol), tris (dibenzylideneacetone) dipalladium (49.12 mg,0.054 mmol) and 2-bicyclohexylphosphine-2 ',6' -diisopropyloxybiphenyl (25.03 mg,0.0536 mmol) 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 methylene chloride (2X 5 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the crude product which was purified by silica gel column (petroleum ether/ethyl acetate=2/8) 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 ethyl ester 047H (47 mg, white solid), yield 33.50%.
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
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 ester 047H (44 mg,0.0701 mmol) was dissolved in methanol with sodium hydroxide (8.82 mg,0.21 mmol): water = 3:1 (2 mL). The reaction mixture was reacted at 40℃for 16 hours. After the reaction was completed, 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, pale yellow solid) in a yield of 64.19%.
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 (30 mg,0.05 mmol) and 4-nitrophenyl-N-methoxycarbamate were dissolved in tetrahydrofuran (2 mL), followed by triethylamine (46 mg,0.45 mmol). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, the mixture was extracted with methylene chloride (2X 35 mL). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give the 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 3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester (Cpd-007)
First step
Preparation of diethyl 2-amino-5- { [ (tert-butoxy) carbonyl ] amino } thiophene-3, 4-dicarboxylate
Diethyl 2, 5-diaminothiophene-3, 4-dicarboxylate 48a (4.00 g,15.49 mmol) was dissolved in dichloromethane (50 mL), followed by the addition of Boc anhydride (3.38 g,15.49 mmol), triethylamine (4.70 g,46.47 mmol), DMAP (189 mg,1.55 mmol). The reaction solution was stirred at room temperature for 2 hours. After LCMS detection, the solvent was evaporated off and the crude product purified by silica gel column (petroleum ether/ethyl acetate=30%) and concentrated under reduced pressure to give the target product diethyl 2-amino-5- { [ (tert-butoxy) carbonyl ] amino } thiophene-3, 4-dicarboxylate 48b (2.00 g, white solid), yield: 25%.
MS m/z(ESI)：359.1[M+1] + 。
Second step
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 (800 mg,2.23 mmol) was dissolved in toluene (20 mL), followed by addition of phenyl chloroformate (1.04 g,6.69 mmol) and stirring at 110℃for 3h. After completion of the reaction, toluene was removed after concentration under reduced pressure. The crude product was purified by silica gel column (petroleum ether/ethyl acetate=20%) and concentrated to give the target product diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ (phenoxycarbonyl) amino ] thiophene-3, 4-dicarboxylate 48c (800 mg, yellow solid), yield: 45%.
MS m/z(ESI)：479.2[M+1] + 。
Third step
Preparation of diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-3, 4-dicarboxylic acid
Diethyl 2- { [ (tert-butoxycarbonyl ] amino } -5- [ (phenoxycarbonyl) amino ] thiophene-3, 4-dicarboxylate 48c (600 mg,1.25 mmol) and 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyaniline 47b (470 mg,1.5 mmol) were dissolved in THF (20 mL), and triethylamine (380 mg,3.75 mmol) was added to the reaction solution. The reaction solution was stirred at room temperature for 16h. After completion of the reaction, the crude product was concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate=30%) 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)。
Fourth step
Preparation of methyl 6- { [ (tert-butoxy) carbonyl ] amino } -3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate
Diethyl 2- { [ (tert-butoxy) carbonyl ] amino } -5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } amino ] thiophene-3, 4-dicarboxylic acid 48d (200 mg,0.29 mmol) was dissolved in methanol (20 mL) followed by sodium methoxide (47 mg,0.87 mmol) 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.) the organic phases were combined and then washed with saturated water, dried over anhydrous sodium sulfate and concentrated to give crude product after purification over a silica gel column (petroleum ether/ethyl acetate=40%) which was concentrated to give methyl 6- { [ (tert-butoxy) carbonyl ] amino } -3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate (70 mg, 48 mg) as a white solid in 36% yield.
MS m/z(ESI)：638.1[M+1] + 。
Fifth step
Preparation of methyl 6-amino-3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-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-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate 48e (30 mg,0.08 mmol) 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 evaporated to give 48f (23 mg, white solid), yield of methyl 6-amino-3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate: 81%.
MS m/z(ESI)：538.1[M+1] + 。
Sixth step
Preparation of 3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester
Preparation 48f (23 mg,0.04 mmol) of methyl 6-amino-3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylate was dissolved in anhydrous tetrahydrofuran (9 mL), followed by addition of phenyl chloroformate (20 mg,0.13 mmol). The reaction was stirred at 90℃for 2h. After completion of the reaction, concentrated under reduced pressure to give 48g (40 mg, brown oil) of 3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester, yield: 71%.
MS m/z(ESI)：562.1[M+1] + 。
Seventh step
Preparation of 3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -6- [ (methoxycarbamoyl) amino ] -2, 4-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester
48g (20 mg,0.03 mmol) of 3- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-6- [ (phenoxycarbonyl) amino ] -1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester was dissolved in THF (10 mL), followed by the addition of triethylamine (63 mg,0.62 mmol) and methylhydroxylamine hydrochloride (26 mg,0.31 mmol). The reaction was stirred at room temperature for 16h. The solvent was evaporated after completion of the reaction 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-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester Cpd-007 (2 mg, white solid), yield: 10%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-008)
First step
Preparation of methyl 5-amino-4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-methoxy-5-nitrobenzoate 064a (1.00 g,4.36 mmol) was dissolved in acetonitrile (30 mL) and saturated ammonium chloride solution (10 mL), followed by iron powder (2.44 g,43.63 mmol). The reaction was stirred at room temperature for 4 hours. After LCMS detection was completed, 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 then the solvent was evaporated to give the target product methyl 5-amino-4-fluoro-2-methoxybenzoate 064b (800 mg, white solid), yield: 87%.
MS m/z(ESI)：200.1[M+1] + 。
Second step
Preparation of methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate
Methyl 5-amino-4-fluoro-2-methoxybenzoate 064B (800 mg,2.23 mmol) was dissolved in dichloromethane (30 mL), followed by the addition of B-anhydride (4.38 g,20.08 mmol), DMAP (49 mg,2.01 mmol) and triethylamine (1.22 mg,12.05 mmol) and stirring at 25℃for 16h. After the reaction was completed, the solvent was removed after concentration under reduced pressure. The crude product was concentrated after passing through a silica gel column (petroleum ether/ethyl acetate=85/15) to give the target product methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate 064c (300 mg, colorless oil), yield: 22%.
MS m/z(ESI)：300.1[M+1] + 。
Third step
Preparation of tert-butyl N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamate
Methyl 5- { [ (tert-butoxy) carbonyl ] amino } -4-fluoro-2-methoxybenzoate 064c (500 mg,1.67 mmol) was dissolved in THF (10 mL) followed by the addition of diisobutylaluminum hydride (2.4 mL,1M in n-hexane) at 0deg.C. The reaction was stirred at room temperature for 2h. Saturated ammonium chloride was added to the mixture to quench the reaction, extracted with ethyl acetate (3×20 mL), dried over anhydrous sodium sulfate, 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 tert-butyl N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamate 064d (230 mg, colorless oil), yield: 46%.
MS m/z(ESI)：294.2[M+1] + 。
Fourth step
Preparation of tert-butyl N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamate
Tert-butyl N- [ 2-fluoro-5- (hydroxymethyl) -4-methoxyphenyl ] carbamate 064d (230 mg,0.85 mmol) was dissolved in dichloromethane (20 mL), followed by addition of carbon tetrabromide (337 mg,1.01 mmol) and triphenylphosphine (267 mg,1.01 mmol). The reaction solution was stirred at 25℃for 16 hours. After completion of the reaction, the crude product was concentrated to give N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 064e (180 mg, white solid), purified by silica gel column (petroleum ether/ethyl acetate=7/3), yield: 57%.
MS m/z(ESI)：356.2[M+1] + 。
Fifth step
Preparation of tert-butyl N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamate
Indole (105 mg,0.90 mmol) was dissolved in tetrahydrofuran (10 mL) and NaH (36 mg,0.90 mmol) was added. After stirring at room temperature for 0.5h, tert-butyl N- [5- (bromomethyl) -2-fluoro-4-methoxyphenyl ] carbamate 064e (150 mg,0.45 mmol) was added. The reaction was stirred at room temperature for 16 hours. After completion of the reaction, sodium hydride was quenched with ice water, then extracted with dichloromethane (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, concentrated to give a crude product, which was purified by silica gel column (petroleum ether/ethyl acetate=1/1) and concentrated to remove the solvent to give tert-butyl N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamate 064f (60 mg, white solid), yield: 34%.
MS m/z(ESI)：371.2[M+1] + 。
Sixth step
Preparation of 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline
Tert-butyl N- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamate 064f (60 mg,0.18 mmol) is dissolved in dichloromethane (9 mL) followed by trifluoroacetic acid (3 mL). The reaction was stirred at 25℃for 2h. After completion of the reaction, concentrated under reduced pressure to give 064g (40 mg, white solid) of 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline, yield: 75%.
MS m/z(ESI)：271.1[M+1] + 。
Seventh step
Preparation of dimethyl 4- ({ [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-5- (indol-1-ylmethyl) -4-methoxyaniline 064g (40 mg,0.15 mmol) was dissolved in tetrahydrofuran (20 mL), and 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester (70 mg,0.19 mmol) and triethylamine (45 mg,0.44 mmol) were added, and the reaction was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, compound 4- ({ [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid dimethyl ester 064h (60 mg, brown solid), yield: 55%.
MS m/z(ESI)：512.2[M+1] + 。
Eighth step
Preparation of 3- [ 2-fluoro-5- (indol-1-ylmethyl) -4-methoxyphenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction solution in the seventh step. Lithium hydroxide monohydrate (11 mg,0.25 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a 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-dioxo-1H-thieno [2,3-d ] pyrimidine-5-carboxylic acid methyl ester Cpd-008 (2 mg, white solid), yield: 4%.
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)
First step
Preparation of methyl 4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-hydroxybenzoate 065a (2 g,11.80 mmol), methyl iodide (3 g,23.60 mmol) and cesium carbonate (4 g,12.98 mmol) were dissolved in acetonitrile (10 mL). The mixture was reacted at 25 ℃ for 6 hours, then extracted with ethyl acetate (3×50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the residue obtained after concentration was separated and purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give methyl 4-fluoro-2-methoxybenzoate 065b (2.1 g, transparent oil), yield: 87%.
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
Preparation of 5-bromo-4-fluoro-2-methoxybenzoic acid methyl ester
Methyl 4-fluoro-2-methoxybenzoate 065b (1.00 g,5.4 mmol), NBS (0.96 g,5.4 mmol), ferric trichloride (90 mg,0.54 mmol) was dissolved in acetonitrile (4 mL) and stirred at 60℃for 1 hour. After the reaction was completed, the residue obtained after concentration was separated and purified by 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%.
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).
Third step
Preparation of 5-bromo-4-fluoro-2-methoxybenzoic acid
5-bromo-4-fluoro-2-methoxybenzoic acid methyl ester 065c (1.00 g,3.80 mm)ol) and lithium hydroxide (45 mg,19.00 mmol) 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 diluted hydrochloric acid, extracted with ethyl acetate (2X 30 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 065d (900 mg, white solid) of 5-bromo-4-fluoro-2-methoxybenzoic acid in 76% yield.
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).
Fourth step
Preparation of 5-bromo-4-fluoro-2-methoxybenzoyl chloride
5-bromo-4-fluoro-2-methoxybenzoic acid 065d (900 mg,3.6 mmol) was dissolved in anhydrous dichloromethane (20 mL) and oxalyl chloride (3.00 g,24 mmol) was added dropwise followed by DMF (2.00 g,28 mmol). The mixture was stirred at 25℃for 0.5 h. After the reaction was completed, the excess solvent and oxalyl chloride were removed by concentration, and the obtained 5-bromo-4-fluoro-2-methoxybenzoyl chloride 065e (900 mg, white solid) was directly used for the next reaction in a yield of 65%.
MS m/z(ESI)：263.0(M-Cl+CH 3 )。
Fifth step
Preparation of 1- [ (5-bromo-4-fluoro-2-methoxyphenyl) carbonyl ] indole
1H-indole (2 g,20.30 mmol) was dissolved in tetrahydrofuran (10 mL). Sodium hydride (1.00 g,27.20 mol) was then added at 0deg.C. The mixture was stirred at 25℃for 1 hour, then 5-bromo-4-fluoro-2-methoxybenzoyl chloride 065e (900 mg,3.4 mmol) was dissolved in tetrahydrofuran (5 mL) and the solution was added dropwise thereto, followed by stirring at 25 ℃. After 1.5 hours, the reaction solution was slowly poured into ice water to quench, and the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, extracted with dichloromethane (2X 30 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: 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).
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 (320 mg,0.92 mmol), benzhydryl amine (66 mg,3.68 mmol), BINAP (114 mg,0.18 mmol), tris (dibenzylideneacetone) dipalladium (84 mg,0.09 mmol) and cesium carbonate (599 mg,1.84 mmol) 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), extracted with dichloromethane (3×50 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and the residue obtained after concentration was purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 065g (220 mg, yellow oil), yield of N- (diphenylmethylene) -2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline: 43%.
MS m/z(ESI)：449.1[M+1] + 。
Seventh step
Preparation of 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline
N- (diphenylmethylene) -2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline 065g (110 mg,0.24 mmol) was dissolved in tetrahydrofuran (5 mL), and concentrated hydrochloric acid (1 mL) was slowly added dropwise thereto, and the mixture was reacted at 25℃for 5 minutes. After the reaction was completed, the pH was adjusted to 7-8 with saturated sodium bicarbonate solution, extracted with ethyl acetate (3×50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the resulting residue was purified on a silica gel column (petroleum ether/ethyl acetate=2/1) to give 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyaniline 065h (80 mg, orange oil), yield: 74%.
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 (40 mg,0.14 mmol) and methyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (236 mg,0.70 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (79 mg,1.41 mmol) 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 methyl 4- [ ({ 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate (50 mg, white solid), yield 34%.
MS m/z(ESI)：526.0[M+1] + 。
Ninth 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 ester
Preparation 065i (50 mg,0.09 mmol) of methyl 4- [ ({ 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate was dissolved in tetrahydrofuran (5 mL), and lithium hydroxide (11 mg,0.47 mmol) 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 completed, extracted with dichloromethane (3×50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, 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), yield: 44%.
MS m/z(ESI)：494.0[M+1] + 。
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
3- { 2-fluoro-5- [ (indol-1-yl) carbonyl ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid methyl ester 065j (35 mg,0.0709 mmol) and trimethyltin hydroxide (64 mg,0.3545 mmol) were dissolved in 1, 2-dichloroethane (4 mL). The mixture was reacted at 80℃for 16 hours. After the reaction was completed, it was prepared by direct concentration and preliminary purification by reverse-phase column (water: acetonitrile=3:2), and after purification, 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) was obtained in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-010)
First step
Preparation of 2- (2-fluoro-4-methoxyphenyl) isoindole-1, 3-dione
2-fluoro-4-methoxyaniline 66a (500 mg,3.54 mmol) was dissolved in N, N-dimethylformamide (10 mL), followed by the addition of phthalic anhydride (577 mg,3.89 mmol). 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 saturated NaCl solution, anhydrous Na 2 SO 4 After drying, filtration and concentration under reduced pressure, 2- (2-fluoro-4-methoxyphenyl) isoindole-1, 3-dione 66b (500 mg, white solid) was obtained, yield: 49%.
MS m/z(ESI)：272.2[M+1] + 。
Second step
Preparation of 5- (1, 3-dioxoisopropyl-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
2- (2-fluoro-4-methoxyphenyl) isoindole-1, 3-dione 66b (200 mg,0.74 mmol) was dissolved in chlorosulfonic acid (3.44 mL,29.49 mmol) and stirred at room temperature for 2h. After the completion of the reaction, the reaction mixture 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, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 5- (1, 3-dioxyisopropyl-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 66c (250 mg, white solid) yield: 64%.
MS m/z(ESI)：392.0[M+1] + 。
Third step
Preparation of 2- { [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid
Indole (95 mg,0.81 mmol) was dissolved in THF (5 mL), then NaH (95 mg,0.81mmol, kerosene retention, content 60%) was added to the reaction solution under ice bath, and after stirring at room temperature for 0.5h, 5- (1, 3-dioxaisopropanol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 66c (150 mg,0.41 mmol) was added. The reaction solution was stirred at room temperature for 4h, water was added after the completion of the reaction, dichloromethane was used for extraction three times, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to give 66d (180 mg, white solid) of 2- { [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid, yield: 85%.
MS m/z(ESI)：469.1[M+1] + 。
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 group]Carbamoyl } benzoic acid 66d (130 mg,0.32 mmol) and sodium acetate (79 mg,0.96 mmol) were dissolved in acetic anhydride (5 mL). The reaction solution was stirred at 90℃for 2 hours. After the reaction was completed, it was cooled to room temperature and then saturated NaHCO 3 Wash and ethyl acetate (3×30 mL) extract. The organic phases are combined, saturated saline water washed, 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%.
MS m/z(ESI)：451.1[M+1] + 。
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 (140 mg,0.31 mmol) was dissolved in ethanol (10 mL), then hydrazine hydrate (1.56 mL,31.08 mmol) 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 saturated 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), yield: 76%.
MS m/z(ESI)：321.1[M+1] + 。
Sixth step
Preparation of dimethyl 4- ({ [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-5- (indole-1-sulfonyl) -4-methoxyaniline 66f (50 mg,0.16 mmol) was dissolved in anhydrous tetrahydrofuran (9 mL), followed by the addition of triethylamine (47 mg,0.47 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (73 mg,0.20 mmol). The reaction was stirred at room temperature for 16h. And directly carrying out the next reaction without post-treatment after the reaction is finished.
MS m/z(ESI)：562.1[M+1] + 。
Seventh step
Preparation of 3- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To 66g of the reaction solution in the sixth step were added MeOH (1 mL) and water (1 mL), followed by addition of lithium hydroxide monohydrate (11 mg,0.25 mmol). The reaction was stirred at room temperature for 2h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparative HPLC (0.1% formic acid/acetonitrile/water) to give Cpd-010 (20 mg, white solid), yield of 3- [ 2-fluoro-5- (indole-1-sulfonyl) -4-methoxyphenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid: 46%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-011)
First step
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxybenzene
4-fluoro-2-methoxyphenol (1.00 g,7.04 mmol) and K 2 CO 3 (1.5 g,10.55 mmol) was dissolved in acetone (10 mL) and benzyl bromide (1.4 g,8.4 mmol) was added. The reaction was carried out at 65℃for 16h. The reaction mixture 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 give preparation 068b of 1- (benzyloxy) -4-fluoro-2-methoxybenzene (1.4 g, white solid), yield: 77%.
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
Preparation of 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene
Compound 068b (500 mg,2.15 mmol) was dissolved in acetic anhydride (3 mL) and nitric acid (65%) (0.2 mL) was slowly added dropwise at 0deg.C. The reaction was carried out at 0℃for 4h. The reaction mixture was quenched with water (10 mL), extracted with dichloromethane (3X 20 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=10:1) to give 1- (benzyloxy) -4-fluoro-2-methoxy-5-nitrobenzene 068c (430 mg, pale yellow solid), yield: 58%.
MS m/z(ESI)：278.1[M+1] + .
Third step
Preparation of 4-fluoro-2-methoxy-5-nitrophenol
Compound 068c (550 mg,1.98 mmol) was dissolved in a solution of boron trichloride in dichloromethane (1M, 5 mL) and reacted at room temperature for 2h. 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), yield: 50%.
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).
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 (500 mg,1 mmol), compound 068d (994.6 mg,5 mmol) and triethylamine (537.8 mg,5 mmol) were dissolved in ethyl acetate (30 mL) and reacted at 80℃for 5 hours. The reaction mixture 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 then boron trifluoride etherate (750 mg,5 mmol) was added. The reaction was carried out at 50℃for 3 hours. The reaction solution was saturated with NaHCO 3 The solution (50 mL) was quenched, 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: 72%.
MS m/z(ESI)：457.1[M+1] + .
Fifth step
Preparation of 2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) aniline
Compound 068e (700 mg,1.53 mmol) was dissolved in methanol (15 mL) and saturated NH 4 Cl (5 mL) solutionTo the mixed solution, iron powder (857 mg,15.3 mmol) was then added. 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 taken up in saturated NaHCO 3 And NaCl solution washing, and drying with anhydrous sodium sulfate. Filtering, concentrating the filtrate under reduced pressure to give 2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) aniline 068f (470 mg, brown solid), yield: 65%.
MS m/z(ESI)：427.2[M+1] + .
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 (420 mg,0.98 mmol), triethylamine (298 mg,2.95 mmol) and dimethyl 2, 3-dimethyl-4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (399mg, 1.18 mmo) were dissolved in THF (20 mL) and reacted at room temperature for 16h. The reaction solution was concentrated under reduced pressure to give dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((1-tolyl-1H-indol-3-yl) oxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 068g (600 mg, brown oily liquid), yield: 55%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 068g (600 mg,0.90 mmol) and water and lithium hydroxide (188 mg,4.49 mmol) were dissolved in THF/MeOH/H 2 O=5:1:1 (20 mL) for 3h at room temperature. Concentrating the reaction solution under reduced pressure, and subjecting the residue to reverse phase column chromatography (ACN: H) 2 O (0.2% FA) =60%) to obtain 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%.
MS m/z(ESI)：622.1[M+1] + .
Eighth step
Preparation of 3- (5- (((1H-indol-3-yl) oxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 068h (100 mg,0.16 mmol) was dissolved in a mixed solution of THF/meoh=2:1 (10 mL), then cesium carbonate (262 mg,0.80 mmol) was added and reacted at room temperature for 16h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by preparative HPLC (ACN: H 2 O(0.1％NH 3 ) =95%) to obtain 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 (10 mg, pale yellow solid), yield: 13%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-012)
First step
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 (1.5 g,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 solution 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 was 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
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 (200 mg,0.58 mmol) 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 solution 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] + 。
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 (70 mg,0.22 mmol) in THF (3 mL) was added triethylamine (226 mg,2.2 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (87 mg,0.56 mmol). The reaction mixture was stirred at 25℃for 16 hours. The reaction mixture was concentrated. The concentrated residue was purified using a flash column to give 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, reddish brown solid). The yield thereof was found to be 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).
Fourth step
Preparation of 3- (5- ((1H-indol-3-yl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To 4- { [ (2-fluoro-4-methoxy-5- { [1- (4-methyl-1-sulfonylphenyl) indol-3-yl)]Amino } phenyl) carbamoyl]Amino } thiophene-2, 3-dicarboxylic acid dimethyl ester 069d (110 mg,0.20 mmol) in THF/MeOH/H 2 To a solution of o=1:1:2 (2 mL) was added lithium hydroxide (10.8 mg,0.45 mmol). The reaction mixture was stirred at 25℃for 20 min. The reaction solution was purified by flash column to obtain crude product. Purifying the crude product by a high performance liquid phase preparation method to obtain a product 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 (1.02 mg, brown solid). Yield: 2.33%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-013)
First step
Preparation of 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene
Tert-butyl nitrite (8.32 g,80.7 mmol) and copper bromide (9.01, 40.3 mmol) were dissolved in acetonitrile (40 mL) under nitrogen atmosphere, the reaction solution was heated to 50℃and then a solution of 4-fluoro-2-methoxy-5-nitroaniline 070a (5 g,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 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), yield: 73%.
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
Preparation of 5-bromo-2-fluoro-4-methoxyaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 070b (2 g,8 mmol) is dissolved in ethanol/water=4/1 (20 mL), iron powder (2.23 g,40 mmol) and ammonium chloride (2.14 g,40 mmol) are 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) in a yield of 81%.
MS m/z(ESI)：220.1[M+1] + .
Third step
Preparation of (5-bromo-2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester
5-bromo-2-fluoro-4-methoxyaniline 070c (1.5 g,6.8 mmol) was dissolved in tetrahydrofuran (20 mL), di-tert-butyl dicarbonate (1.78 g,8.1 mmol) and triethylamine (2.06 g,20.4 mmol) were added, and the reaction 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 give the title product (tert-butyl 5-bromo-2-fluoro-4-methoxyphenyl) carbamate 070d (1.3 g, yellow solid) in a yield of 56%.
MS m/z(ESI)：342.0[M+1] + .
Fourth step
Preparation of tert-butyl [ 2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl ] carbamate
Tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate 070d (1.2 g,3.7 mmol) was dissolved in dioxane (15 mL) under nitrogen, 4, 5-tetramethyl-2- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) -1,3, 2-dioxaborane (1.41 g,5.5 mmol), 1' -bis (diphenylphosphino) ferrocene palladium dichloride (0.27 g,0.3 mmol) and potassium acetate (0.73 g,7.4 mmol) were added and the reaction stirred at 90℃for 8 hours. The residue obtained by concentrating the reaction solution was purified by a silica gel column (petroleum ether/ethyl acetate=5/1) to give the title product [ tert-butyl 2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl ] carbamate 070e (1.1 g, yellow solid) in a yield of 75%.
MS m/z(ESI)：390.2(M+23).
Fifth step
Preparation of [5- (dihydroxyborane) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester
Tert-butyl [ 2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl ] carbamate 070e (1.1 g,3 mmol) was dissolved in tetrahydrofuran (20 mL) and water (10 mL), sodium periodate (1.93 g,9 mmol) and ammonium chloride (0.48 g,9 mmol) 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 silica gel column (petroleum ether/ethyl acetate=9/1) to give the title product [5- (dihydroxyborane) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 070f (0.55 g, yellow solid), yield: 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).
Sixth step
Preparation of tert-butyl {5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamate
1-benzofuran-3-carbaldehyde (256 mg,1.75 mmol) was dissolved in toluene (5 mL) under a nitrogen atmosphere, tert-butyl [5- (dihydroxyborane) -2-fluoro-4-methoxyphenyl ] carbamate 070f (500 mg,1.75 mmol), 2- [ bis (2, 4-di-tert-butyl-phenoxy) phosphino ] -3, 5-di (tert-butyl) phenyl-palladium (II) chloride dimer (138 mg,0.09 mmol) and potassium phosphate (742 mg,3.5 mmol) were added and the reaction was stirred at 50℃for 6 hours. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate=2/1) to give 070g (40 g, colorless oil) of the title product {5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamic acid tert-butyl ester, yield: 6%.
MS m/z(ESI)：410.1(M+23).
Seventh step
Preparation of [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester
Dibutyl {5- [ 1-benzofuran-3-yl (hydroxy) methyl ] -2-fluoro-4-methoxyphenyl } carbamic acid tert-butyl ester 070g (40 mg,0.11 mmol) was dissolved in dichloromethane (2 mL) under nitrogen atmosphere, triethylsilane (24 mg,0.21 mmol) was added under ice-water bath, and after stirring for 10 minutes trifluoroacetic acid (36 mg,0.31 mmol) was added and the reaction solution stirred at 25℃for 1 hour. To the reaction solution was added water (50 mL), followed by extraction 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 [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamic acid tert-butyl ester 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 (35 mg,0.1 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (215 mg,1.88 mmol) was added and the reaction was stirred at 25℃for 1 h. The concentrated residue was purified by silica gel column (petroleum ether/ethyl acetate=3/2) to give the title product 5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyaniline 070i (25 mg, yellow oil) in 78% yield.
MS m/z(ESI)：272.2[M+1] + .
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 (30 mg,0.11 mmol) was dissolved in tetrahydrofuran (3 mL), dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (37 mg,0.11 mmol) and triethylamine (34 mg,0.33 mmol) were added sequentially, the reaction was stirred at 25℃for 16 hours, and the reaction was concentrated to give the title product dimethyl 4- ({ [5- (1-benzofuran-3-ylmethyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate 070j (40 mg, yellow oil) in 56% yield.
MS m/z(ESI)：513.0[M+1] + .
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-dicarboxylic acid 070j (40 mg,0.08 mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (17 mg,0.39 mmol) was added, and the reaction mixture was stirred at 25℃for 2 hours. Water (0.5 mL) was then added and the reaction stirred at 25℃for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution, followed by extraction 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)
First step
Preparation of 1-bromo-4-fluoro-2-methoxy-5-nitrobenzene
Cuprous bromide (5.79 g,0.04 mol) and t-butyl nitrite (8.32 g,0.08 mol) were dissolved in acetonitrile (20 mL), stirred at 50℃for 10 minutes, and a solution of 4-fluoro-2-methoxy-5-nitroaniline (5.00 g,0.03 mol) in acetonitrile (30 mL) was added to the reaction solution, which was stirred at 50℃for 2 hours. After completion of the reaction, the reaction mixture was quenched with 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 separated and purified by silica gel column (petroleum ether/ethyl acetate=9/1) to give 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
Preparation of 5-bromo-2-fluoro-4-methoxyaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 071b (4.88 g,0.02 mol), iron powder (2.18 g,0.04 mmol) and ammonium chloride (2.09 g,0.04 mmol) were dissolved in ethanol: water = 1:1 (20 mL), the reaction was allowed to react at 25℃for 1 hour. After completion of the reaction, the reaction mixture was quenched with 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=7/3) to give 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)。
Third step
Preparation of (5-bromo-2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester
5-bromo-2-fluoro-4-methoxyaniline 071c (2 g,9.1 mmol) and di-tert-butyl dicarbonate (2.98 g,13.6 mmol) were dissolved in tetrahydrofuran (20 mL), triethylamine (1.84 g,18.2 mmol) was added at room temperature, and the reaction solution was reacted at 60℃for 16 hours. After completion of the reaction, the reaction mixture was quenched with 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 with silica gel column (petroleum ether/ethyl acetate=15/1) to give 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)。
Fourth step
Preparation of tert-butyl (2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl) carbamate
Tert-butyl (5-bromo-2-fluoro-4-methoxyphenyl) carbamate 071d (438 mg,1.36 mmol), 4, 5-tetramethyl-2- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) -1,3, 2-dioxaborane (418 mg,1.63 mmol), 1' -bis (diphenylphosphino) ferrocene palladium dichloride (100 mg,0.14 mmol) and potassium acetate (267 mg,2.7 mmol) were dissolved in 1, 4-dioxane (10 mL) and the reaction was reacted at 90℃for 10 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (3X 20 mL). The resulting residue was purified by silica gel column (petroleum ether/ethyl acetate=9/1) to give tert-butyl (2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl) carbamate 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)。
Fifth step
Preparation of (5- ((tert-butoxycarbonyl) amino) -4-fluoro-2-methoxyphenyl) boronic acid
Tert-butyl (2-fluoro-4-methoxy-5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) phenyl) carbamate 071e (345 mg,0.93 mmol), sodium periodate (601 mg,2.8 mmol) and ammonium chloride (150 mg,0.08 mmol) were dissolved in tetrahydrofuran: water = 3:1 (20 mL), the reaction was allowed to react at room temperature for 48 hours. After completion of the reaction, the reaction mixture was quenched with 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)。
Sixth step
Preparation of (5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester
(5- ((tert-Butoxycarbonyl) amino) -4-fluoro-2-methoxyphenyl) boronic acid 071f (200 mg,0.7 mmol), 1-benzothiophene-3-carbaldehyde (170 mg,1.05 mmol), 2- [ bis (2, 4-di-tert-butyl-phenoxy) phosphino ] -3, 5-di (tert-butyl) phenyl-palladium (II) chloride dimer (55 mg,0.03 mmol) and potassium phosphate (298 mg,1.39 mmol) were dissolved in toluene (10 mL) and the reaction solution was reacted at 50℃for 2 hours under nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched with 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 silica gel column (petroleum ether/ethyl acetate=1/1) to give 071g (75 mg, yellow solid) of (5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamic acid tert-butyl ester in 25% yield.
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
Tert-butyl (5- (benzo [ b ] thiophen-3-yl (hydroxy) methyl) -2-fluoro-4-methoxyphenyl) carbamate 071g (65 mg,0.13 mmol), triethylsilane (19 mg,0.16 mmol) and trifluoroacetic acid (45 mg,0.34 mmol) were dissolved in dichloromethane (10 mL), and the reaction solution was reacted at room temperature under nitrogen atmosphere for 10 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL) and extracted with methylene chloride (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting 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 (10 mg,0.14 mmol) and dimethyl 2, 3-dimethyl-4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (56 mg,0.16 mmol) were dissolved in tetrahydrofuran (8 mL), triethylamine (28 mg,0.28 mmol) was added at room temperature, the reaction mixture was reacted at room temperature for 16 hours, water (20 mL) was added to quench the reaction mixture after the reaction was completed, extracted with ethyl acetate (3×20 mL), all the organic phases were combined, dried over anhydrous sodium sulfate and concentrated, and the resulting residue was purified with a silica gel column (petroleum ether/ethyl acetate=7/3) to give methyl (4- (3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 071i (31 mg, yellow solid) in 40% yield.
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)。
Ninth step
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
Dimethyl 4- (3- (5- (benzo [ b ] thiophen-3-ylmethyl) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 071i (25 mg,0.05 mmol) and lithium hydroxide (11 mg,0.5 mmol) 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 completion of the reaction, 1N hydrochloric acid was added to the reaction mixture to quench the mixture to pH <7, followed by extraction with ethyl acetate (3X 10 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified with reverse phase column (47% acetonitrile/water) to give 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 (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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-015)
First step
Preparation of methyl 4-fluoro-2-methoxybenzoate
Methyl 4-fluoro-2-hydroxybenzoate 072a (3.00 g,17.63 mmol) was dissolved in acetonitrile (30 mL) followed by the addition of methyl iodide (3.75 g,26.45 mmol) and cesium carbonate (11.49 g,35.26 mmol). The reaction solution was stirred at room temperature for 16 hours. After LCMS detection reaction was completed, dichloromethane (3×100 mL) was added for extraction, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and then the solvent was evaporated off and passed through a silica gel column (petroleum ether/ethyl acetate=6/4) to give the target product methyl 4-fluoro-2-methoxybenzoate 072b (3.00 g, anhydrous oil), yield: 83%.
MS m/z(ESI)：185.2[M+1] + 。
Second step
Preparation of methyl 4-fluoro-2-methoxy-5-nitrobenzoate
Methyl 4-fluoro-2-methoxybenzoate 072b (1.00 g,5.43 mmol) was dissolved in concentrated sulfuric acid (4 mL), followed by the addition of concentrated nitric acid (0.5 mL) under ice-bath. Stirred at 0deg.C for 0.5h. After completion of the reaction, the reaction mixture was poured into ice, a saturated sodium carbonate solution was added to adjust the pH to 7-8, 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 the solvent was removed. The crude product was concentrated after passing through a silica gel column (petroleum ether/ethyl acetate=30/70) to give the target product methyl 4-fluoro-2-methoxy-5-nitrobenzoate 072c (700 mg, white solid), yield: 53%.
MS m/z(ESI)：230.2[M+1] + 。
Third step
Preparation of 4-fluoro-2-methoxy-5-nitrobenzoic acid
Methyl 4-fluoro-2-methoxy-5-nitrobenzoate 072c (300 mg,1.75 mmol) was dissolved in THF (10 mL) and water (2 mL), followed by sodium hydroxide (262 mg,5.23 mmol). The reaction was stirred at room temperature for 1h. After the reaction was completed, 1N hydrochloric acid was added to adjust the pH to 4-5, extracted with dichloromethane (3×20 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), yield: 72%.
MS m/z(ESI)：216.2[M+1] + 。
Fourth step
Preparation of 4-fluoro-2-methoxy-5-nitrobenzoyl chloride
4-fluoro-2-methoxy-5-nitrobenzoic acid 072d (100 mg,0.46 mmol) was dissolved in dichloromethane (10 mL), followed by the addition of oxalyl chloride (118 mg,0.93 mmol) and DMF (0.1 mL). The reaction solution was stirred at 25℃for 1 hour. After completion of the reaction, concentrated to give 4-fluoro-2-methoxy-5-nitrobenzoyl chloride 072e (120 mg, white solid), yield: 88%.
MS m/z(ESI)：230.2(M-Cl+CH 3 )。
Fifth step
Preparation of 3- [ (4-fluoro-2-methoxy-5-nitrophenyl) carbonyl ] -1H indole
4-fluoro-2-methoxy-5-nitrobenzoyl chloride 072e (120 mg,0.51 mmol) and aluminum trichloride (137 mg,1.02 mmol) were dissolved in dichloromethane (15 mL) followed by the addition of indole (120 mg,1.02 mmol). 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 after filtration to give the 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%.
MS m/z(ESI)：315.2[M+1] + 。
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 (120 mg,0.38 mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by iron powder (213 mg,3.82 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 mL), and 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%.
MS m/z(ESI)：285.2[M+1] + 。
Seventh step
Preparation of dimethyl 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyaniline 072g (50 mg,0.18 mmol) was dissolved in tetrahydrofuran (10 mL), and 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester (71 mg,0.21 mmol) and triethylamine (53 mg,0.53 mmol) were added. The reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, the compound 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 072H (100 mg, pale yellow liquid), yield: 54%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction solution in the seventh step. Lithium hydroxide monohydrate (12 mg,0.28 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product, which was purified by preparation (0.1% formic acid/acetonitrile/water) to give dimethyl 4- [ ({ 2-fluoro-5- [ (1H-indol-3-yl) carbonyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid Cpd-015 (20 mg, white solid), yield: 43%.
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)
First step
1-tosyl-1H-indole-7-carbaldehyde
Sodium hydride (124 mg,5.17 mmol) and indole-7-carbaldehyde 78a (500 mg,3.44 mmol) were added to tetrahydrofuran (20 mL) under ice-bath conditions, and after stirring at room temperature for 20 minutes, p-toluenesulfonyl chloride (985 mg,5.17 mmol) 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 with 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 resulting residue was separated by silica gel column chromatography, eluting with n-hexane/ethyl acetate (4:1 (v/v)) to give 1-tosyl-1H-indole-7-carbaldehyde 78b (700 mg, white solid) in 65% yield.
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
(1-tosyl-1H-indol-7-yl) methanol
1-tosyl-1H-indole-7-carbaldehyde 78b (700 mg,2.32 mmol) was dissolved in MeOH (10 mL), and sodium borohydride (26 mg,6.96 mmol) was added thereto, and the reaction was stirred at 25℃for 2 hours. After the 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 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).
Third step
7- (bromomethyl) -1-tosyl-1H-indole
(1-tosyl-1H-indol-7-yl) methanol 78c (520 mg,1.76 mmol) is dissolved in dichloromethane (6 mL), to which tetrabromomethane (23 mg,0.07 mmol) and triphenylphosphine (18 mg,0.07 mmol) are added. 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 silica gel column (petroleum ether/ethyl acetate=4/1) afforded 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).
Fourth step
7- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1-tosyl-1H-indole
7- (bromomethyl) -1-toluenesulfonyl-1H-indole 78d (430 mg,1.16 mmol) was dissolved in 5mL of acetonitrile, to which 4-fluoro-2-methoxy-5-nitrophenol (220 mg,1.18 mmol) and potassium carbonate (195 mg,1.41 mmol) were added. 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).
Fifth step
2-fluoro-4-methoxy-5- ((1-toluenesulfonyl-1H-indol-7-yl) methoxy) aniline
7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1- (4-methyl-1-sulfonylphenyl) indole 78e (500 mg,1 mmol) was dissolved in MeOH (5 mL), and palladium on carbon (50 mg) was added thereto, and the reaction system was placed in a hydrogen balloon atmosphere and the air in the reaction system was replaced. After the reaction was completed, the mixture was filtered off palladium on carbon, the filter cake (3×30 mL) was washed with methanol, and the organic phase was concentrated to give 2-fluoro-4-methoxy-5- ((1-tosyl-1H-indol-7-yl) methoxy) aniline 78f (300 mg, pale yellow solid) in 60% yield. Directly used in the next reaction.
MS m/z(ESI)：441.1[M+1] + 。
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
2-fluoro-4-methoxy-5- ((1-toluenesulfonyl-1H-indol-7-yl) methoxy) aniline 78f (300 mg,1 mmol) was added to a solution of THF (5 mL), and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (522 mg,1.53 mmol) and triethylamine (1.05 g,10.35 mmol) were added, and the reaction solution was stirred at room temperature for 12 hours. After the reaction was completed, 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, dried over anhydrous sodium sulfate and concentrated to give the crude product which was further purified by 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 (250 mg,0.36 mmol) was dissolved in a solvent (tetrahydrofuran/methanol=2/1), 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 (3×30 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 purified by HPLC (mobile phase: acetonitrile/water) 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%.
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)
First step
Preparation of 8-hydroxymethylquinoline
8-Aldoloquinoline (1 g,6.36 mmol) was dissolved in methanol (10 mL) and NaBH was added 4 (720 mg,3 mmol). 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, gray solid), yield: 71%.
MS m/z(ESI)：160.2[M+1] + .
Second step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline
4-fluoro-2-methoxy-5-nitrophenol (500 mg,2.67 mmol), 8-hydroxymethylquinoline (425.3 mg,2.67 mmol) and PPh3 (911.1 mg,3.47 mmol) were dissolved in dry THF (10 mL) and DIAD (702.4 mg 3.47 mmol) was added. The reaction was carried out at 45℃for 2 hours. 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), yield: 39%
MS m/z(ESI)：329.1[M+1] + .
Third step
Preparation of 2-fluoro-4-methoxy-5- (quinoline-8-methoxy) aniline
Compound 080c (370 mg,1.13 mmol) was dissolved in MeOH (10 mL) with saturated NH 4 To a mixed solution of Cl (2 mL) was then added iron powder (629 mg,11.27 mmol). The reaction was carried out at room temperature for 2 hours, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (20 mL) and taken up in saturated NaHCO 3 And NaCl solution washing, and drying with 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), yield: 88%. Yield: 44%.
MS m/z(ESI)：299.2[M+1] + .
Fourth step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- (quinolin-8-ylmethoxy) phenyl) ureido) thiophene-2, 3-dicarboxylate
Compound 080d (320 mg,1.07 mmol) and triethylamine (326 mg,3.22 mmol) were dissolved in THF (10 mL) and then dimethyl 4- ((phenoxycarbonyl) amino) thiophene dimethyl-2, 3-dicarboxylic acid dimethyl ester (432 mg,1.2872 mmol) was added. After 16h reaction at room temperature, the reaction mixture was concentrated under reduced pressure to give dimethyl 4- (3- (2-fluoro-4-methoxy-5- (quinolin-8-ylmethoxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 080e (450 mg, yellow oily liquid), yield: 62%.
MS m/z(ESI)：540.2[M+1] + .
Fifth step
Preparation of (3- (2-fluoro-4-methoxy-5- (quinoline-8-methoxy) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 080e was dissolved in (450 mg,0.83 mmol) in THF/MeOH/H 2 To a mixed solution of o=5:1:1 (30 mL), then lithium hydroxide (105 mg,2.50 mmol) was added. The reaction was carried out at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the 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-dioxo-1H-thieno [3, 4-d) ]Pyrimidine-5-carboxylic acid Cpd-017 (60 mg, pale yellow solid), yield: 14.6%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-018)
First step
Preparation of 5- (bromomethyl) quinoxaline
5-methylquinoxaline 081a (2.20 g,0.02 mol) was dissolved in carbon tetrachloride (30 mL), N-bromosuccinimide (2.72 g,0.02 mol) and a catalytic amount of benzoyl peroxide were added, and reacted 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, filtered, 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%.
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
Preparation of 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline
5- (bromomethyl) quinoxaline 081b (142 mg,0.64 mmol) was dissolved in acetonitrile (10 mL), potassium carbonate (106 mg,0.76 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (119 mg,0.64 mmol) were added, and reacted at 70℃for 3 hours. After the reaction was completed, water (10 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 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 081c (200 mg, white solid), yield: 95%.
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).
Third step
Preparation of 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline
5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 081c (257 mg,0.78 mmol) was dissolved in ethanol (3 mL) and water (3 mL), iron powder (131 mg,2.34 mmol) and ammonium chloride (125 mg,2.34 mmol) were added, and the reaction solution was reacted at 80℃for 1 hour. After the reaction was completed, extraction with ethyl acetate (3×20 mL), drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate and purification by column chromatography (mobile phase: petroleum ether/ethyl acetate=4/1) gave 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline 081d (143 mg, white solid), yield: 61%.
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).
Fourth step
Preparation of dimethyl 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) aniline 081d (113 mg,0.38 mmol) was dissolved in tetrahydrofuran (5 mL), triethylamine (57 mg,0.57 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (152 mg,0.45 mmol) were added, the reaction was stirred at 50℃for 4 hours after completion of the reaction, the reaction solution was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=1/1) to give dimethyl 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate 081e (184 mg, white solid) in 90% yield.
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).
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
Dimethyl 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinoxalin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 081e (134 mg,0.25 mmol) was dissolved in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL), lithium hydroxide (31 mg,0.74 mmol) was added, and the mixture was reacted at 25℃for 2 hours. After the reaction was completed, diluted 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 via a 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%.
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)
First step
Preparation of 5-methyl quinazoline
To a solution of 5-bromoquinazoline 083a (1.00 g,4.80 mmol) in N, N-dimethylformamide (20 mL) was added methylboronic acid (340 mg,5.70 mmol), 1' -bis (diphenylphosphino) ferrocene palladium dichloride (350 mg,0.40 mmol) and potassium carbonate (710 mg,7.20 mmol). The reaction mixture was stirred in a microwave apparatus at 80℃for 1 hour. After the reaction was completed, water was added to quench, extraction was performed with ethyl acetate (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=3/1) to give 5-methylquinazoline 083b (299 mg, white solid), yield: 44%.
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
Preparation of 5- (bromomethyl) quinazoline
To a solution of 5-methylquinazoline 083b (299 mg,2.07 mmol) in carbon tetrachloride (10 mL) were added N-bromosuccinimide (447 mg,2.49 mmol) and 2,2' -azobis (2-methylpropanenitrile) (34 mg,0.21 mmol), and the reaction mixture was stirred under nitrogen at 85℃for 3 hours. After the reaction was completed, water was added to quench, extracted with dichloromethane (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) to give 5- (bromomethyl) quinazoline 083c (290 mg, white solid), yield: 63%.
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).
Third step
Preparation of 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinazoline
To a solution of 4-fluoro-2-methoxy-5-nitrophenol (292 mg,1.56 mmol) in acetonitrile was added potassium carbonate (270 mg,1.95 mmol) and 5- (bromomethyl) quinazoline 083c (290 mg,1.30 mmol), and the reaction mixture was stirred at 70℃for 2 hours. After the reaction was completed, water quenching was added, extraction was performed with methylene chloride (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: methylene chloride/methanol=20/1) to give 5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinazoline 083d (317 mg, yellow solid), yield: 74%.
MS m/z(ESI)：330.0[M+1] + 。
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,0.87 mmol) in ethanol (5 mL) and water (5 mL) were added iron powder (146 mg,2.61 mmol) and ammonium chloride (140 mg,2.61 mmol), and the reaction mixture was stirred at 80℃for 1 hour. After the reaction was completed, the filtrate was filtered while it is hot, extracted with methylene chloride (3×20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated and dried to give 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) aniline 083e (217 mg, yellow solid), yield: 83%.
MS m/z(ESI)：300.1[M+1] + 。
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 (167 mg,0.56 mmol) in tetrahydrofuran (10 mL) was added triethylamine (85 mg,0.84 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (225 mg,0.67 mmol), the reaction mixture was stirred at 25 ℃ for 8 hours, after the reaction was completed, water was added, quenched, extracted with dichloromethane (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=1/1) to give dimethyl 4- ({ [ [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate 083f (200 mg, white solid), yield: 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).
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 (170 mg,0.31 mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (23 mg,0.94 mmol), and the mixture was reacted at 25℃for 2 hours. After completion of the reaction, 2N 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 via a preparative column to give Cpd-019 (56 mg, white solid) 3- [ 2-fluoro-4-methoxy-5- (quinazolin-5-ylmethoxy) phenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid, yield: 36%.
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)
First step
Preparation of 2- (2-fluoro-4-methoxyphenyl) isoindole-1, 3-dione
2-fluoro-4-methoxyaniline 085a (5.00 g,35.40 mmol) and 2-benzofuran-1, 3-dione (16.00 g,106.20 mmol) were dissolved in DMF. The mixture was reacted in a microwave oven at 120 ℃ for 1 hour, then quenched with water (50 mL) and extracted with ethyl acetate (3×50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and the residue obtained after concentration was separated and purified by 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), yield: 55%.
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
Preparation of 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
2- (2-fluoro-4-methoxyphenyl) isoindole-1, 3-dione 085b (3.00 g,11.00 mmol) was dissolved in dichloromethane (3 mL) and slowly added dropwise to chlorosulfonic acid (30 mL) and after stirring at 25℃for 4 hours, the reaction mixture was slowly poured into ice water and quenched and extracted with ethyl acetate (3X 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) in 60% yield.
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).
Third step
Preparation of 2- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] isoindole-1, 3-dione
3-methyl-1H-indole (552 mg,4.06 mmol) was dissolved in tetrahydrofuran (10 mL). Sodium hydride (195 mg,8.11 mmol) was then added at 0deg.C. The mixture was stirred at 0℃for 0.5 hours, then 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 085c (300 mg,0.81 mmol) was dissolved in tetrahydrofuran (5 mL), the solution was added dropwise, and stirring was continued at 25 ℃. After 1.5 hours, the reaction solution was slowly poured into ice water to quench, and the pH was adjusted to 6-7 with 1M dilute hydrochloric acid, extracted with dichloromethane (2X 30 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: 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)。
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 (140 mg,0.30 mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The mixture was stirred at 90℃for 1 hour and then dried by spin-drying to remove the solvent, and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 2/3) to give 085e (80 mg, orange solid) of 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) aniline in 63% yield.
MS m/z(ESI)：335.0[M+1] + 。
Fifth step
Preparation of dimethyl 4- ({ [ [ [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
Preparation 085e (30 mg,0.09 mmol) of 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) aniline and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (72 mg,0.21 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (36 mg,0.36 mmol) was added and the mixture was stirred at 25℃for 16 hours.
MS m/z(ESI)：576.0[M+1] + 。
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 addition of lithium hydroxide (10 mg,0.43 mmol). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, the mixture was directly concentrated and preliminarily purified by a reverse column (water: acetonitrile=3:2) followed by preparation (acetonitrile/0.1% aqueous formic acid 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 as prepared Cpd-020 (2.09 mg, white solid) in a yield of 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)
First step
Preparation of 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride
2H-1, 4-benzoxazine (79 mg,0.6 mmol) and triethylamine (164 mg,1.60 mmol) were dissolved in acetonitrile (5 mL). 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 086a (200 mg,0.5 mmol) was then added. The reaction solution 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
Preparation of 5- (2, 3-dihydro-1, 4-benzoxazine-4-sulfonyl) -2-fluoro-4-methoxy aniline
5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride 086b (210 mg, 0.4478 mmol) and hydrazine hydrate (1 mL) are dissolved in ethanol (5 mL). The reaction solution 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).
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 (80 mg,0.24 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (237 mg,0.71 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (71 mg,0.71 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：580.1[M+1] + 。
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 of the third step were added methanol (3 mL) and water (1 mL), followed by addition of lithium hydroxide (6 mg,0.14 mmol). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=60:40) followed by preparation (acetonitrile/0.1% aqueous formic acid solution) to give 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) in 52% yield.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-022)
First step
Preparation of 3, 4-dihydro-2H-quinoxaline-1-carboxylic acid tert-butyl ester
1,2,3, 4-tetrahydroquinoxaline 087a (500 mg,3.72 mmol) was dissolved in dichloromethane (20 mL) and then triethylamine (1.13 g,11.18 mmol), N, N-dimethylpyridin-4-amine (45 mg,0.37 mmol) and B-anhydride (813 mg,3.72 mmol) were added. The reaction solution was stirred at 25℃for 3 hours. After the 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, pale yellow oil), yield: 33%.
MS m/z(ESI)：257.1(M+Na)。
Second step
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 (700 mg,2.99 mmol) was dissolved in acetonitrile (3 mL), methyl iodide (826 mg,5.97 mmol) and potassium carbonate (823 mg,5.97 mmol) were then added, stirring was carried out at 80℃for 16 hours, water (10 mL) was added to the reaction solution after completion of the reaction, extraction was carried out three times with methylene chloride, the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate and concentrated to give a crude product, which was purified by silica gel column (petroleum ether/ethyl acetate=85/15) to give 4-methyl-2, 3-dihydroquinoxaline-1-carboxylic acid tert-butyl ester 087c (550 mg, brown oil), yield: 38%.
MS m/z(ESI)：271.1(M+Na)。
Third step
Preparation of 1-methyl-3, 4-dihydro-2H-quinoxaline
4-methyl-2, 3-dihydroquinoxaline-1-carboxylic acid tert-butyl ester 087c (300 mg,1.21 mmol) was dissolved in dichloromethane (5 mL), followed by addition of trifluoroacetic acid (1 mL). The reaction solution was stirred at 25℃for 2 hours. After completion of LCMS monitoring reaction, preparation 087d (200 mg, brown oil), yield, of 1-methyl-3, 4-dihydro-2H-quinoxaline was obtained after concentration under reduced pressure: 89%.
MS m/z(ESI)：149.2[M+1] + 。
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 (250 mg,0.10 mmol) and 5- (1, 3-dioxaisopropanol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (686 mg,1.86 mmol) were dissolved in 1, 2-dichloroethane (10 mL), followed by DIPEA (264 mg,5.06 mmol) and pyridine (267 mg,3.37 mmol). The reaction solution was stirred at 100℃for 1 hour. After completion of the reaction, water (10 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 a crude product, which was passed through a silica gel column (petroleum ether/ethyl acetate=4/6) to give 2- [ 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) phenyl ] isoindole-1, 3-dione 087e (200 mg, white solid), yield: 12%.
MS m/z(ESI)：482.2[M+1] + 。
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 (100 mg,0.22 mmol) was dissolved in ethanol (5 mL) followed by the 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, 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 a crude product, which was subjected to a silica gel column (petroleum ether/ethyl acetate=3/7) to give 30g (50 mg, white solid) of 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) aniline, yield: 63%.
MS m/z(ESI)：374.2[M+1] + 。
Sixth step
Preparation of dimethyl 4- ({ [ 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) phenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
30g (50 mg,0.14 mmol) of 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) aniline was dissolved in tetrahydrofuran (20 mL), and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (57 mg,0.17 mmol) and triethylamine (43 mg,0.43 mmol) were added to stir the reaction solution at 25℃for 16 hours. After completion of LCMS monitoring the reaction, this step was used directly in the next step without work-up.
MS m/z(ESI)：593.1[M+1] + 。
Ninth step
Preparation of 3- [ 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) phenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To the reaction solution of the sixth step were added (2 mL) and methanol (5 mL), lithium hydroxide (24 mg,0.56 mmol) was added, and the reaction solution was stirred at 25℃for 3 hours. After LCMS monitoring the reaction to completion, the reaction was concentrated to remove the organic solvent and the residue was prepared (acetonitrile/water/0.1% formic acid) to give 3- [ 2-fluoro-4-methoxy-5- (4-methyl-2, 3-dihydroquinoxaline-1-sulfonyl) phenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-022 (8 mg, orange solid), yield: 17%.
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)
First step
Preparation of 2- [5- (4-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1, 3-dione
4-chloro-1H-indole 088a (135 mg,0.89 mmol) was dissolved in tetrahydrofuran (5 mL). Sodium hydride (58 mg,2.43 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 0.5 hours. Then, 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (300 mg,0.81 mmol) was dissolved in tetrahydrofuran (2 mL), and the solution was added dropwise thereto, and the reaction mixture was reacted at 25℃for 3 hours. The pH was then adjusted to 6-7 with 1M dilute hydrochloric acid, dichloromethane (2X 10 mL) was used for extraction, 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
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 (89 mg,0.18 mmol) was dissolved in acetic acid (2 mL). Sodium acetate (43 mg,0.53 mmol) was then added. The reaction mixture was reacted at 90℃for 1.5 hours. Then, the pH was 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] + 。
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 (80 mg,0.16 mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction solution was reacted at 90℃for 4 hours. The solvent was then dried by spinning 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] + 。
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 (80 mg,0.23 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (90 mg,0.27 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (68 mg,0.67 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：596.0[M+1] + 。
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 of the fourth step were added methanol (3 mL) and water (1 mL), followed by addition of lithium hydroxide (5 mg,0.14 mmol). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=60:40) followed by preparation (acetonitrile/0.1% aqueous formic acid solution) to give 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), yield 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)
First step
Preparation of 8-bromo-5-chloroisoquinoline
5-chloroisoquinoline 089a (1 g,6.1 mmol) was dissolved in sulfuric acid (15 mL), N-bromosuccinimide (1.19 g,6.7 mmol) was added under ice-water bath, and the reaction solution was stirred at 25℃for 16 hours. The reaction was quenched with water (100 mL), adjusted to pH 8 with 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
Preparation of 5-chloroisoquinoline-8-carboxylic acid methyl ester
8-bromo-5-chloroisoquinoline 089b (1.5 g,6.2 mmol) was dissolved in methanol (30 mL) and 1,1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride dichloromethane complex (0.51 g,0.6 mmol) and sodium acetate (0.76 g,9.3 mmol) were added. The reaction solution was stirred at 60℃for 8 hours under 10 atmospheres of pressure under a carbon monoxide atmosphere. The residue obtained by concentrating the reaction solution was purified by silica gel column (petroleum ether/ethyl acetate=4/1) to give the title product methyl 5-chloroisoquinoline-8-carboxylate 089c (1.1 g, white solid) in 75% yield.
MS m/z(ESI)：222.2[M+1] + .
Third step
Preparation of (5-chloroisoquinolin-8-yl) methanol
Methyl 5-chloroisoquinoline-8-carboxylate 089c (500 mg,2.26 mmol) was dissolved in tetrahydrofuran (10 mL) methanol (2 mL), and sodium borohydride (256 mg,6.77 mmol) was added under ice water bath. The reaction solution was stirred at 25℃for 16 hours. The reaction was quenched with water (100 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. 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] + .
Fourth step
Preparation of 5-chloro-8- (chloromethyl) isoquinoline
(5-chloroisoquinolin-8-yl) methanol 089d (400 mg,2.07 mmol) 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), yield: 71%.
MS m/z(ESI)：212.1[M+1] + .
Fifth step
Preparation of 5-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
4-fluoro-2-methoxy-5-nitrophenol (106 mg,0.57 mmol) was dissolved in N, N-dimethylformamide (3 mL), and 5-chloro-8- (chloromethyl) isoquinoline 089e (120 mg,0.57 mmol), potassium carbonate (157 mg,1.13 mmol) and potassium iodide (94 mg,0.57 mmol) were added. The reaction solution was stirred at 80℃for 2 hours. The reaction was quenched with water (50 mL) 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 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] + .
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 (90 mg,0.25 mmol) was dissolved in ethanol (4 mL) and water (1 mL), iron powder (70 mg,1.24 mmol) and ammonium chloride (67 mg,1.24 mmol) were added the reaction solution was stirred at 90℃for 6 hours, filtration was carried out, and the residue obtained by concentrating the filtrate was purified by 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 dimethyl 4- [ ({ 5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 089g (20 mg,0.06 mmol) was dissolved in tetrahydrofuran (3 mL), dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (20 mg,0.06 mmol) and triethylamine (19 mg,0.18 mmol) were added sequentially, and the reaction solution was stirred at 25℃for 16 hours.
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
Dimethyl 4- [ ({ {5- [ (5-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid ester 089h (30 mg,0.05 mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (11 mg,0.26 mmol) was added, and the reaction solution was stirred at 25℃for 2 hours. Water (0.5 mL) was then added and the reaction stirred at 25℃for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution, followed by extraction 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)
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First step
Preparation of [ (2-bromo-5-fluorophenyl) methyl ] (2, 2-dimethoxyethyl) amine
A solution of 2-bromo-5-fluorobenzaldehyde 090a (5.00 g,0.02 mol) and 2, 2-dimethoxyethylamine (3.88 g,0.04 mol) in ethanol (30 mL) was stirred at 80℃for 8 hours, then sodium cyanoborohydride (1.55 g,0.02 mol) and a catalytic amount of acetic acid (1 mL) were added and stirred at 25℃for 1 hour. After completion of the reaction, quench with saturated aqueous ammonium chloride, extract with ethyl acetate (3×50 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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), yield: 72%.
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
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 (5.18 g,0.02 mol) and triethylamine (2.69 g,0.03 mol) in methylene chloride (30 mL) was added dropwise p-toluenesulfonyl chloride (3.71 g,0.02 mol) at 0℃and reacted at 0℃for 3 hours. After completion of the reaction, quench with water (20 mL), extract with ethyl acetate (3×50 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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%.
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).
Third step
Preparation of 8-bromo-5-fluoroisoquinoline
To a solution of aluminum trichloride (6.75 g,0.06 mol) in methylene chloride (10 mL), a solution of [ (2-bromo-5-fluorophenyl) methyl ] (2, 2-dimethoxyethyl) [ (4-sulfonylphenyl) methyl ] amine 090c (5.00 g,0.01 mol) in methylene chloride (10 mL) was added dropwise at 0℃and after completion of the addition, the reaction was carried out at 0℃for 8 hours. After the reaction was completed, the reaction solution was slowly dropped to ice water for quenching, and extracted with methylene chloride (3×50 mL). The organic phase was washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, 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), yield: 45%.
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).
Fourth step
Preparation of 5-fluoroisoquinoline-8-carboxylic acid methyl ester
To a solution of 8-bromo-5-fluoroisoquinoline 090d (1.13 g,5.00 mmol) and triethylamine (1.52 g,15.00 mmol) in methanol (10 mL) was added [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) (0.37 g,0.50 mmol) and reacted at 80℃under 1atm carbon monoxide for 8 hours. After the reaction was completed, water (10 mL) was added thereto, 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), yield: 46%.
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).
Fifth step
Preparation of (5-fluoroisoquinolin-8-yl) methanol
To a solution of methyl 5-fluoroisoquinoline-8-carboxylate 090e (400 mg,1.95 mmol) in methanol (5 mL) was added sodium borohydride (221 mg,5.85 mmol) at 0deg.C and the reaction was carried out at 0deg.C for 8 hours. After completion of the reaction, a saturated aqueous ammonium chloride solution (5 mL) was added dropwise thereto to quench the reaction mixture, and the mixture was 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), yield: 26%.
MS m/z(ESI)：178.1[M+1] + 。
Sixth step
Preparation of 8- (bromomethyl) -5-fluoroisoquinoline
To a solution of (5-fluoroisoquinolin-8-yl) methanol 090f (90 mg,0.51 mmol) in methylene chloride (3 mL) was added dropwise phosphine tribromide (138 mg,0.51 mmol) at 0℃and the reaction was carried out at 0℃for 2 hours. After the reaction was completed, a saturated aqueous sodium hydrogencarbonate solution was added to adjust ph=6 to 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: 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 (62 mg,0.33 mmol) and potassium carbonate (69 mg,0.50 mmol) in acetonitrile (5 mL) was added 090g (80 mg,0.33 mmol) of 8- (bromomethyl) -5-fluoroisoquinoline, and the mixture was reacted at 25℃for 8 hours. After completion of the reaction, water (5 mL) was added thereto, followed by extraction 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), yield: 87%.
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 (80 mg,0.23 mmol) in ethanol (2 mL) and water (2 mL) was added iron powder (39 mg,0.69 mmol) and ammonium chloride (37 mg,0.69 mmol) and reacted at 80℃for 1 hour. After completion of the reaction, extracted with ethyl acetate (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyaniline 090i (56 mg, yellow solid), yield: 77%.
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).
Ninth step
Preparation of dimethyl 4- [ ({ 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
To a solution of 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyaniline 090i (46 mg,0.15 mmol) and triethylamine (44 mg,0.44 mmol) in tetrahydrofuran (3 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (59 mg,0.17 mmol), and after completion of the reaction at 25℃for 3 hours, water (5 mL) was added to quench, extraction with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and 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).
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-dicarboxylic acid 090j (78 mg,0.14 mmol) in tetrahydrofuran (1 mL), methanol (1 mL) and water (1 mL) was added lithium hydroxide monohydrate (18 mg,0.42 mmol), and the mixture was reacted 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 via a preparative column to give Cpd-025 (11 mg, white solid) as 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid, yield: 16%.
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)
First step
Preparation of 5- (bromomethyl) quinoxaline
5-methylquinoxaline 091a (2.20 g,0.02 mol) was dissolved in carbon tetrachloride (30 mL), N-bromosuccinimide (2.72 g,0.02 mol) and a catalytic amount of benzoyl peroxide were added and reacted 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, filtered, and concentrated to give 5- (bromomethyl) quinoxaline 091b (2.80 g, yellow solid), after purification by column chromatography (mobile phase: 20% ethyl acetate/50% petroleum ether), yield: 82%.
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
Preparation of 4-fluoro-2-methoxy-5-nitro-N- (quinoxalin-5-ylmethyl) aniline
5- (bromomethyl) quinoxaline 091b (1.70 g,7.60 mmol) was dissolved in acetonitrile (20 mL), potassium carbonate (1.26 g,9.10 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (1.41 g,7.60 mmol) were added and reacted at 70℃for 3 hours. After the completion of the reaction, water (10 mL) was added to quench, and solids were precipitated and filtered, and the obtained cake was dried to obtain 4-fluoro-2-methoxy-5-nitro-N- (quinoxalin-5-ylmethyl) aniline 091c (1.75 g, brown solid), yield: 70%.
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).
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 (1.70 g,5.20 mmol) was dissolved in ethanol (5 mL) and water (5 mL), iron powder (0.87 g,15.60 mmol) and ammonium chloride (0.83 g,15.60 mmol) were added, and the reaction solution was reacted at 80℃for 1 hour. After the reaction was completed, extracted with ethyl acetate (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=1/1) to give 4-fluoro-6-methoxy-1-N- (quinoxalin-5-ylmethyl) benzene-1, 3-diamine 091d (728 mg, yellow solid), yield: 46%.
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).
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 (6278 mg,2.11 mmol) was dissolved in tetrahydrofuran (5 mL), triethylamine (639 mg,6.32 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (847 mg,2.53 mmol) were added, and after the reaction was stirred at 50℃for 5 hours, the reaction mixture was concentrated and purified by column chromatography (mobile phase: 60% ethyl acetate/40% petroleum ether) to give dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (quinoxalin-5-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate 091e (1.02 g, yellow solid) in a yield of 90%.
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).
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
Dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (quinoxalin-5-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 091e (200 mg,0.37 mmol) was dissolved in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL), lithium hydroxide (47 mg,1.11 mmol) was added, and the mixture was reacted at 25℃for 4 hours. After the reaction was completed, diluted hydrochloric acid was added to adjust 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 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), yield: 24%.
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)
First step
Preparation of 2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid
Sodium hydride (132 mg,2.43 mmol) and 6-fluoroindole (151 mg,0.97 mmol) 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 (300 mg,0.81 mmol) was added. The mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was quenched with water (5 mL) and extracted with ethyl acetate (2X 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=4/1) to give 2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid 092b (158 mg, yellow solid) in 40% yield.
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
Preparation of 2- (2-fluoro-5- (6-fluoro-1H-indol-1-yl) sulfanilamide) -4-methoxybenzene) iso-1, 3-dione
2- ((2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid 092b (158 mg,0.32 mmol) and sodium acetate (79 mg,0.97 mmol) were dissolved in acetic anhydride (5 mL) and the reaction was reacted at 90℃for 2 hours. After completion of the reaction, the reaction mixture was quenched with water (5 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 separated and purified by silica gel column (petroleum ether/ethyl acetate=4/1) to give 2- (2-fluoro-5- (6-fluoro-1H-indol-1-yl) sulfa-2-methoxy-benzene) -iso-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)。
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) sulfa-2-yl) -4-methoxybenzene) isovitamin-1, 3-dione 092c (113 mg,0.24 mmol) 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 with water (5 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=4/1) to give 2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 092d (63 mg, yellow solid) in 77% yield.
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)。
Fourth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
22-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 092d (63 mg,0.18 mmol) and dimethyl 4- ((phenoxycarbonyl) amino) thienyl-2, 3-dicarboxylate (124 mg,0.37 mmol) were dissolved in tetrahydrofuran (5 mL), and triethylamine (37 mg,0.37 mmol) was added thereto, and the reaction solution was reacted at room temperature for 16 hours. After completion of the reaction, the reaction mixture was quenched with water (5 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=1/1) to give dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 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)。
Fifth step
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
Dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 092e (63 mg,0.11 mmol) and lithium hydroxide (13 mg,0.54 mmol) 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, 1M hydrochloric acid (5 mL) was added to the reaction mixture to quench the reaction mixture, and the mixture was extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. Purification of the resulting residue by HPLC preparation (mobile phase: acetonitrile/water) gave 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 (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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-028)
First step
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 (250 mg,0.68 mmol) and 5-chloro-1H-indole (123 mg,0.81 mmol) were dissolved in dry THF (5 mL), sodium hydride (24 mg,1.01 mmol) was added and reacted at 25℃for 3 hours. After completion of the reaction, quench with water (5 mL), extract with ethyl acetate (3×20 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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), yield: 56%.
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
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 (184 mg,0.38 mmol) was dissolved in ethanol (5 mL), and hydrazine hydrate (38 mg,0.76 mmol) was added and reacted at 90℃for 1 hour. After completion of the reaction, quench with water (10 mL), extract with ethyl acetate (3×20 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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), yield: 49%.
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).
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 (60 mg,0.17 mmol), dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (68 mg,0.20 mmol) and triethylamine (51 mg,0.51 mmol) in tetrahydrofuran was stirred at 50℃for 3 hours after completion of the reaction, quenched with water (50 mL) and extracted with ethyl acetate (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate concentrated and purified by column chromatography (mobile phase: dichloromethane/methanol=10/1) to give dimethyl 4- ({ [5- (5-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate 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).
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-dicarboxylic acid 093d (70 mg,0.12 mmol) was dissolved in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL), and lithium hydroxide monohydrate (15 mg,0.35 mmol) was added and reacted at 25℃for 2 hours. After the reaction was completed, diluted 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 a 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), yield: 16%.
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)
First step
Preparation of 2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid
6-chloro-1H-indole 094a (250 mg,1.65 mmol) was dissolved in tetrahydrofuran (5 mL). Sodium hydride (118 mg,4.95 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 0.5 hours. Then, 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (319 mg,1.64 mmol) was dissolved in tetrahydrofuran (2 mL), and the above solution was added dropwise thereto, and the reaction mixture was reacted at 25℃for 3 hours. The pH was then adjusted to 6-7 with 1M dilute hydrochloric acid, dichloromethane (2X 10 mL) was used for extraction, 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 (150 mg, white solid) of 2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid 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
Preparation of 2- [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1, 3-dione
2- { [5- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid 094b (150 mg,0.298 mmol) was dissolved in acetic acid (2 mL). Sodium acetate (73 mg,0.895 mmol) was then added. The reaction mixture was reacted at 90℃for 1.5 hours. Then, the pH was 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- (6-chloroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1, 3-dione 094c (80 mg, white solid) in 50% yield.
MS m/z(ESI)：485.0[M+1] + 。
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 (62 mg,0.13 mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction solution was reacted at 90℃for 4 hours. The solvent was then dried by spinning 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] + 。
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 (30 mg,0.085 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (34 mg,0.27 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (25 mg,0.25 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：596.0[M+1] + 。
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-dicarboxylic acid 094e (5 mL stock solution) and lithium hydroxide (6 mg,0.14 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=60:40) to give 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) after preliminary purification in 7% yield.
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)
First step
Preparation of 2- [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] isoindole-1, 3-dione
5-fluoro-1H-indole 095a (60 mg,0.44 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (32 mg,1.33 mmol) was added under ice-water bath, the reaction was stirred at 0℃for 15 minutes, then 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (165 mg,0.44 mmol) was added, and the reaction was stirred at 25℃for 4 hours. The reaction was quenched with water (50 mL) and then 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
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 (200 mg,0.42 mmol) was dissolved in ethanol (5 mL) and hydrazine hydrate (214 mg,4.27 mmol) was added. The reaction solution was stirred at 90℃for 1 hour. The residue obtained by concentrating the reaction solution was purified by 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), yield: 18%.
MS m/z(ESI)：339.1[M+1] + .
Third step
Preparation of dimethyl 4- ({ [ [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyaniline 095c (30 mg,0.09 mmol) was dissolved in tetrahydrofuran (3 mL), dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (30 mg,0.09 mmol) and triethylamine (27 mg,0.27 mmol) were sequentially added, and the reaction solution was stirred at 25℃for 16 hours.
MS m/z(ESI)：580.1[M+1] + .
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
Dimethyl 4- ({ [ [ 2-fluoro-5- (5-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 095d (30 mg,0.05 mmol) was dissolved in tetrahydrofuran (2 mL), lithium hydroxide monohydrate (11 mg,0.26 mmol) was added, and the reaction solution was stirred at 25℃for 2 hours. Water (0.5 mL) was then added and the reaction stirred at 25℃for an additional 2 hours. The pH was adjusted to 6 with 1N hydrochloric acid solution, followed by extraction 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, pale 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)
First step
Preparation of 2- ((2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) carbamoyl) benzoic acid
7-fluoro-1H-indole 096a (300 mg,2.2 mmol) was dissolved in dry tetrahydrofuran (5 mL) and cooled to 0deg.C under nitrogen. Sodium hydride (300 mg,11 mmol) was then added and the mixture stirred for 30 minutes. Then 5- (1, 3-dioxa-polyol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (410 mg,1.1 mmol) was added thereto, and the reaction solution was stirred at room temperature for 2 hours. After the completion of the reaction, the reaction mixture 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. Purification of the residue by chromatography (methanol/dichloromethane=0-10%) afforded 2- { [ 2-fluoro-5- (7-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid 096b (260 mg, yellow solid), yield: 35%.
MS m/z(ESI)：487.1[M+1] + 。
Second step
Preparation of 2- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) isoindole-1, 3-dione
2- { [ 2-fluoro-5- (7-fluoroindole-1-sulfonyl) -4-methoxyphenyl ] carbamoyl } benzoic acid 096b (160 mg,0.3 mmol) was dissolved in acetic anhydride (5 mL), followed by sodium acetate (134 mg,1.6 mmol) in 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%. The crude product was used directly in the next reaction.
MS m/z(ESI)：491.0(M+23)。
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 (140 mg,0.34 mmol) was dissolved in ethanol (6 mL), and then hydrazine hydrate (68 mg,0.36 mmol) was added to the solution. The resulting mixture 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. Purification of the residue by chromatography (ethyl acetate/petroleum ether=0-20%) afforded 2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 096d (85 mg, brown solid), yield: 63%.
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).
Fourth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyaniline 096d (85 mg,0.25 mmol) is dissolved in tetrahydrofuran (5 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (343 mg,1.0 mmol) and triethylamine (52 mg,0.51 mmol). The reaction solution was stirred at room temperature for 16 hours. After the completion of the reaction, 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 dimethyl 4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 096e (60 mg, brown solid), yield: 42%. The crude product was used directly in the next step.
MS m/z(ESI)：580.1[M+1] + 。
Fifth step
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
Dimethyl 4- (3- (2-fluoro-5- ((7-fluoro-1H-indol-1-yl) sulfonyl) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 096e (60 mg,0.10 mmol) was dissolved in tetrahydrofuran/methanol/water=1: 1:1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. 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), yield: 53%.
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)
First step
Preparation of 2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid
4-fluoro-1H-indole 098a (250 mg,1.85 mmol) was dissolved in tetrahydrofuran (5 mL) followed by sodium hydride (133 mg,5.55 mmol) at 0deg.C. The reaction solution was reacted at 0℃for 0.5 hours. Then, 5- (1, 3-dioxoisoindol-2-yl) -4-fluoro-2-methoxybenzenesulfonyl chloride (684 mg,1.85 mmol) was dissolved in tetrahydrofuran (2 mL), and the above solution was added dropwise thereto, and the reaction mixture was reacted at 25℃for 3 hours. The pH was then adjusted to 6-7 with 1M dilute hydrochloric acid, dichloromethane (2X 10 mL) was used for extraction, 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 (230 mg, white solid) of 2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid in 25% yield.
MS m/z(ESI)：487.0[M+1] + 。
Second step
Preparation of 2- [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] isoindole-1, 3-dione
2- { [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } benzoic acid 098b (200 mg,0.41 mmol) was dissolved in acetic acid (2 mL). Sodium acetate (101 mg,1.23 mmol) was then added. The reaction mixture was reacted at 90℃for 1.5 hours. Then, the pH was 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] + 。
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 (150 mg,0.32 mmol) and hydrazine hydrate (1 mL) were dissolved in ethanol (5 mL). The reaction solution was reacted at 90℃for 4 hours. The solvent was then dried by spinning 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).
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 (30 mg,0.088 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (35 mg,0.10 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (26 mg,0.26 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The reaction solution was not subjected to any post-treatment to obtain dimethyl 4- ({ [5- (4-fluoroindole-1-sulfonyl) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 098e as a raw solution.
MS m/z(ESI)：580.0[M+1] + 。
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
Methanol is added into the reaction liquid in the fourth step: water = 3:1 (4 mL) and lithium hydroxide (3 mg,0.078 mmol). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=60:40) to give (acetonitrile/0.1% aqueous formic acid solution) after preliminary purification 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), 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)
First step
Preparation of [ (2-bromo-3, 5-difluorophenyl) methyl ]2, 2-dimethoxyethyl) amine
2-bromo-3, 5-difluorobenzaldehyde 099a (5.00 g,22.60 mmol) was dissolved in ethanol (50 mL). 2, 2-Dimethoxyethylamine (2.61 g,24.80 mmol) was then added at 25 ℃. The reaction mixture was reacted at 80℃for 16 hours. The solvent was then dried by spin-drying, extracted with ethyl acetate (2X 50 mL), the organic phases combined, 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) in 73% yield.
MS m/z(ESI)：310.0[M+1] + 。
Second step
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 (5.70 g,1.86 mmol) was dissolved in dichloromethane (60 mL). Triethylamine (5.64 g,55.80 mmol) and 4-toluenesulfonyl chloride (10.64 g,55.80 mmol) were then added at 0deg.C. The reaction solution was reacted at 0℃for 1 hour. Then water (2X 50 mL), the organic phases were combined, 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).
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 (1.50 g,3.20 mmol) was dissolved in dichloromethane (10 mL) and added dropwise to a solution of anhydrous aluminum trichloride (2.55 g,19.20 mmol) in dichloromethane (5 mL) at 0deg.C. The reaction mixture was reacted at 25℃for 16 hours. Then quenched with ice 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).
Fourth step
Preparation of 5, 7-difluoro isoquinoline-8-carboxylic acid methyl ester
8-bromo-5, 7-difluoroisoquinoline 099d (460 mg,1.88 mmol) was dissolved in methanol (10 mL). Triethylamine (578mg, 5.65 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (137 mg,0.19 mmol) were then added, and the reaction mixture was pressurized and sealed using a carbon monoxide-filled canister and reacted at 80℃for 16 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 55/45) to give methyl 5, 7-difluoroisoquinoline-8-carboxylate 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).
Fifth step
Preparation of (5, 7-difluoroisoquinolin-8-yl) methanol
Methyl 5, 7-difluoroisoquinoline-8-carboxylate 099e (100 mg,0.45 mmol) was dissolved in tetrahydrofuran (5 mL). Lithium aluminum hydride (85 mg,2.24 mmol) was then added at 0deg.C, and the reaction was allowed to react at 0deg.C for 0.5 hours. Ice water was then added to quench and extracted with dichloromethane (2×5 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).
Sixth step
Preparation of 8- (bromomethyl) -5, 7-difluoroisoquinoline
(5, 7-Difluoroisoquinolin-8-yl) methanol 099f (70 mg,0.35 mmol) was dissolved in dichloromethane (5 mL). Triphenylphosphine (188 mg,0.71 mmol) and carbon tetrabromide (237 mg,0.71 mmol) were then added and the reaction was reacted at 45℃for 16 hours. The concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 099g (52 mg, brown solid) of 8- (bromomethyl) -5, 7-difluoroisoquinoline 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 (70 mg,0.27 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (55 mg,0.30 mmol) were dissolved in acetonitrile (10 mL). Potassium carbonate (196 mg,0.28 mmol) was then added and the reaction was allowed to react at 45℃for 16 hours. The solvent was then dried by spinning 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 (20 mg,0.04 mmol) 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 was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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] + 。
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 (30 mg,0.10 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (42 mg,0.12 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (31 mg,0.31 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：529.1[M+1] + 。
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
Preparation method
Dimethyl 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 (3 mg,0.078 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=82:18) to give 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) in a yield of 6%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-034)
First step
Preparation of isoquinoline-8-carboxylic acid methyl ester
8-bromoisoquinoline 100a (1.80 g,8.65 mmol) was dissolved in methanol (30 mL) followed by addition of Pd (dppf) Cl 2 (633 mg,0.87 mmol) and triethylamine (2.63 g,25.95 mmol). The reaction solution was stirred for 16 hours at 80℃under the protection of carbon monoxide. After LCMS detection reaction was completed, the organic solvent was distilled off under reduced pressure and passed through a silica gel column (petroleum ether/ethyl acetate=6/4) to give the target product isoquinoline-8-carboxylic acid methyl ester 100b (1.00 g, white solid), yield: 56%.
MS m/z(ESI)：188.1[M+1] + 。
Second step
Preparation of 2-oxo-8- (methoxycarbonyl) isoquinoline
Isoquinoline-8-carboxylic acid methyl ester 100b (1.50 g,8.01 mmol) was dissolved in chloroform (10 mL) followed by the addition of m-chloroperoxybenzoic acid (2.77 g,16.03 mmol) under ice-bath. Stirring for 3h at room temperature. After completion of the reaction, a small amount of saturated sodium carbonate solution was added, the sodium carbonate solution layer was extracted with methylene chloride (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, and after filtration, the solvent was removed after concentration under reduced pressure, the objective product 2-oxo-8- (methoxycarbonyl) isoquinoline 100c (600 mg, brown solid) was obtained, yield: 33%.
MS m/z(ESI)：204.0[M+1] + 。
Third step
Preparation of methyl 1-chloroisoquinoline-8-carboxylate
2-oxo-8- (methoxycarbonyl) isoquinoline 100c (450 mg,2.21 mmol) was dissolved in phosphorus oxychloride (8 mL). The reaction solution was stirred at 105℃for 4h. After completion of the reaction, the solvent was dried, saturated sodium carbonate solution was added to adjust the pH to 7-8, followed by extraction with dichloromethane (3×20 mL), drying over anhydrous sodium sulfate, and concentration under reduced pressure gave methyl 1-chloroisoquinoline-8-carboxylate 100d (200 mg, brown oil), yield: 37%.
MS m/z(ESI)：222.2[M+1] + 。
Fourth step
Preparation of (1-chloroisoquinolin-8-yl) methanol
Methyl 1-chloroisoquinoline-8-carboxylate 100d (300 mg,1.35 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of diisobutylaluminum hydride (10.8 mL,10.83mmol,1M in n-hexane) 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), and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to remove the solvent, and purified over a silica gel column (petroleum ether/ethyl acetate=1/1) to give (1-chloroisoquinolin-8-yl) methanol 100e (120 mg, white solid), yield: 44%.
MS m/z(ESI)：194.2[M+1] + 。
Fifth step
Preparation of 8- (bromomethyl) -1-chloroisoquinoline
(1-chloroisoquinolin-8-yl) methanol 100e (100 mg,0.52 mmol) was dissolved in dichloromethane (10 mL), followed by addition of carbon tetrabromide (205 mg,0.62 mmol) and triphenylphosphine (163 mg,0.62 mmol). The reaction was stirred at room temperature for 1 h. After the reaction was completed, the solvent was evaporated and concentrated to give a crude product, which was passed through a silica gel column (petroleum ether/ethyl acetate=6/4) and concentrated to give 8- (bromomethyl) -1-chloroisoquinoline 100f (120 mg, white solid), yield: 82%.
MS m/z(ESI)：256.0[M+1] + 。
Sixth step
Preparation of 1-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline
8- (bromomethyl) -1-chloroisoquinoline 100f (100 mg,0.39 mmol) was dissolved in acetonitrile (20 mL), followed by the addition of 4-4-fluoro-2-methoxy-5-nitrophenol (80 mg,0.43 mmol), potassium carbonate (108 mg,0.78 mmol) and potassium iodide (6 mg,0.04 mmol). 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, and the solvent was distilled off under reduced pressure by filtration to give 100g (150 mg, pale yellow solid) of 1-chloro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) isoquinoline after purification of the crude product through a silica gel column (petroleum ether/ethyl acetate=7/3), yield: 85%.
MS m/z(ESI)：363.2[M+1] + 。
Seventh step
Preparation of 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
100g (120 mg,0.33 mmol) 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 the addition of iron powder (185 mg,3.31 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 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 for 100h (100 mg, pale pink solid), yield: 73%.
MS m/z(ESI)：333.1[M+1] + 。
Eighth step
Preparation of dimethyl 4- [ ({ 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline (50 mg,0.15 mmol) was dissolved in tetrahydrofuran (10 mL), and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (60 mg,0.18 mmol) and triethylamine (46 mg,0.45 mmol) were added. The reaction solution was stirred at 25℃for 16 hours. After completion of the reaction, LCMS monitored the reaction to give compound 4- [ ({ 5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 100i (100 mg, pale yellow liquid), yield: 54%.
MS m/z(ESI)：574.1[M+1] + 。
Ninth step
Preparation of 3- {5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Methanol (2 mL) and water (1 mL) were added to the reaction solution in the seventh step. Lithium hydroxide monohydrate (11 mg,0.26 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (0.1% formic acid/acetonitrile/water) to give 3- {5- [ (1-chloroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-034 (20 mg, pale yellow solid), yield: 42%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-035)
First step
Preparation of 6-fluoro-3, 4-dihydro-2H-1, 4-benzoxazine
To a solution of 2-amino-4-fluorophenol 102a (1.00 g,7.90 mmol) and potassium carbonate (2.18 g,15.80 mmol) in N, N-dimethylformamide (10 mL) was added 1, 2-dibromoethane (1.78 g,9.40 mmol), and the mixture was reacted at 80℃for 8 hours. After completion of the reaction, quench with water (10 mL), extract with ethyl acetate (3×30 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify by column chromatography (mobile phase: petroleum ether/ethyl acetate=10/1) to give 6-fluoro-3, 4-dihydro-2H-1, 4-benzoxazine 102b (178 mg, yellow oil), yield: 15%.
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
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 (90 mg,0.59 mmol) and potassium carbonate (122 mg,0.88 mmol) in acetonitrile (5 mL) was added 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (155 mg,0.59 mmol) and reacted at 70℃for 8 hours. After completion of the reaction, quench with water (5 mL), extract with ethyl acetate (3×30 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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%.
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).
Third step
Preparation of 2-fluoro-5- [ (6-fluoro-2, 3-dihydro-1, 4-benzoxazin-4-yl) methyl ] -4-methoxy aniline
To a solution of 6-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2, 3-dihydro-1, 4-benzoxazine 102c (190 mg,0.57 mmol) in ethanol (5 mL) and water (5 mL) was added iron powder (95 mg,1.69 mmol) and ammonium chloride (91 mg,1.69 mmol), and the mixture was reacted at 80℃for 1 hour. After the reaction was completed, ethyl acetate was added to extract (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, 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), yield: 72%.
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).
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 (114 mg,0.37 mmol) and triethylamine (56 mg,0.56 mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (150 mg,0.45 mmol), and the reaction was carried out at 25℃for 3 hours.
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).
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
To a solution of 4- [ ({ 2-fluoro-5- [ (6-fluoro-2, 3-dihydro-1, 4-benzoxazin-4-yl) methyl ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 102e (100 mg,0.18 mmol) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL) was added lithium hydroxide monohydrate (23 mg,0.55 mmol), and the mixture was stirred at 25℃for 2 hours. After the reaction was completed, diluted 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 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%.
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)
First step
Preparation of 5-fluoro-3, 4-dihydro-2H-1, 4-benzoxazine
To a solution of 2-amino-3-fluorophenol 103a (1.00 g,7.90 mmol) and potassium carbonate (2.18 g,15.80 mmol) in N, N-dimethylformamide (10 mL) was added 1, 2-dibromoethane (1.78 g,9.40 mmol), and the mixture was reacted at 80℃for 8 hours. After the reaction was completed, quenched with water (10 mL), extracted with ethyl acetate (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=10/1) to give 5-fluoro-3, 4-dihydro-2H-1, 4-benzoxazine 103b (262 mg, yellow oil), yield: 22%.
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
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 (90 mg,0.59 mmol) and potassium carbonate (122 mg,0.88 mmol) in acetonitrile (5 mL) was added 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (155 mg,0.59 mmol) and reacted at 70℃for 8 hours. After completion of the reaction, quench with water (5 mL), extract with ethyl acetate (3×30 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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), yield: 92%.
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).
Third step
Preparation of 2-fluoro-5- [ (5-fluoro-2, 3-dihydro-1, 4-benzoxazin-4-yl) methyl ] -4-methoxy aniline
To a solution of 5-fluoro-4- [ (4-fluoro-2-methoxy-5-nitrophenyl) methyl ] -2, 3-dihydro-1, 4-benzoxazine 103c (181 mg,0.54 mmol) in ethanol (5 mL) and water (5 mL) was added iron powder (90 mg,1.61 mmol) and ammonium chloride (86 mg,1.61 mmol), and the mixture was reacted at 80℃for 1 hour. After the reaction was completed, ethyl acetate was added to extract (3×30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 2-fluoro-5- [ (5-fluoro-2, 3-dihydro-1, 4-benzoxazin-4-yl) methyl ] -4-methoxyaniline 103d (150 mg, yellow oil), yield: 91%.
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).
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 (140 mg,0.46 mmol) and triethylamine (56 mg,0.56 mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (184 mg,0.55 mmol), and the reaction was carried out at 25℃for 3 hours.
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
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-dicarboxylic acid 103e (100 mg,0.18 mmol) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL) was added lithium hydroxide monohydrate (23 mg,0.55 mmol), and the mixture was stirred at 25℃for 2 hours. After the reaction was completed, diluted 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 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%.
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)
First step
Preparation of 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene
(4-fluoro-2-methoxy-5-nitrophenyl) methanol 104a (60 mg,0.30 mmol), triphenylphosphine (156 mg,0.60 mmol) and carbon tetrabromide (198mg, 0.60 mmol) were dissolved in dichloromethane (5 mL). The mixture was stirred at 25 ℃ for 2 hours and concentrated, and the residue obtained was purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene 104b (50 mg, white solid), yield: 56%.
MS m/z(ESI)：264.0[M+1] + 。
Second step
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 (50 mg,0.19 mmol), 3, 4-dihydro-2H-1, 4-benzoxazine (51 mg,0.38 mmol) and triethylamine (61 mg,1.51 mmol) were dissolved in acetonitrile (5 mL), the mixture was stirred at 90 ℃ for 1 hour and concentrated, and the resulting residue was isolated and purified by 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), yield: 46%.
MS m/z(ESI)：319.1[M+1] + 。
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 (35 mg,0.11 mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (308 mg,5.50 mmol) and stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated to give 5- (2, 3-dihydro-1, 4-benzoxazine-4-methyl) -2-fluoro-4-methoxyaniline 104d (25 mg, brown oil). The yield thereof was found to be 47%.
MS m/z(ESI)：289.1[M+1] + 。
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 (25 mg,0.09 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (145 mg,0.43 mmol) were dissolved in tetrahydrofuran (5 mL) then triethylamine (48 mg,0.87 mmol) was added and the mixture was stirred at 25℃for 16 hours.
MS m/z(ESI)：530.1[M+1] + 。
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 of the fourth step were added methanol (3 mL) and water (1 mL), followed by addition of lithium hydroxide (9 mg,0.38 mmol). The mixture was reacted at 25℃for 90 minutes. After the reaction was completed, the mixture was directly concentrated and purified by reverse column (water: acetonitrile=3:2) followed by preparation (acetonitrile/0.1% aqueous formic acid solution) to give 3- [ 2-fluoro-4-methoxy-5- (3-methylindole-1-sulfonyl) phenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid as prepared Cpd-037 (2.10 mg, orange solid) in 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)
First step
Preparation of 2, 3-difluoro-6-methoxyaniline
2, 3-difluoro-6-methoxybenzoic acid 105a (700 mg,3.72 mmol) was dissolved in N, N-dimethylformamide (20 mL). Triethylamine (560 mg,5.58 mmol) and diphenyl azide phosphate (560 mg,5.58 mmol) were added sequentially. 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
Preparation of 2-amino-3, 4-difluorophenol
2, 3-difluoro-6-methoxyaniline 105b (580 mg,3.64 mmol) was dissolved in dichloromethane (5 mL) and boron tribromide (4.57 g,18.22 mmol) was added at 0deg.C. The reaction solution 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] + 。
Third step
Preparation of 5, 6-difluoro-3, 4-dihydro-2H-1, 4-benzoxazine
2-amino-3, 4-difluorophenol 105c (500 mg,3.45 mmol) was dissolved in N, N-dimethylformamide (5 mL). Then, potassium carbonate (2.59 g,10.34 mmol) and 1, 2-dibromoethane (777 mg,4.13 mmol) were successively added, and the reaction solution was reacted at 85℃for 16 hours. After completion of the reaction, water (2 mL) was added thereto, 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] + 。
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 (120 mg,0.70 mmol) and 1- (bromomethyl) -4-fluoro-2-methoxy-5-nitrobenzene (222 mg,0.84 mmol) were dissolved in N, N-dimethylformamide (5 mL). Then, potassium carbonate (290 mg,2.10 mmol) and potassium iodide (8 mg, 0.004mmol) were added, and the reaction mixture was reacted at 65℃for 16 hours. After completion of the reaction, water (2 mL) was added thereto, 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 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] + 。
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 (120 mg,0.34 mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (189 mg,3.39 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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] + 。
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-dicarboxylic acid
5- [ (5, 6-difluoro-2, 3-dihydro-1, 4-benzoxazin-4-yl) methyl ] -2-fluoro-4-methoxyaniline 105f (60 mg,0.19 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (124 mg,0.37 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (56 mg,0.56 mmol) was then added, and the reaction was allowed to react at 25℃for 1 hour. The reaction solution was not subjected to any post-treatment to obtain 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-dicarboxylic acid.
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
109g (5 mL of stock solution) of 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 was dissolved with lithium hydroxide (13 mg,0.53 mmol) in methanol: water=3:1 (4 mL.) after the reaction was reacted at 25℃for 90 minutes, after the reaction was completed, direct concentration was performed by reverse column (water: acetonitrile=30:70) and purification was performed after preliminary purification to give 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 (13.24 mg, yellow solid), yield: 13.46%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-039)
First step
Preparation of isoquinolin-8-yl methanol
Isoquinoline-8-carbaldehyde 107a (300 mg,1.91 mmol) was dissolved in anhydrous methanol (10 mL) and then sodium borohydride (22 mg,0.57 mmol) was added under ice-bath. The reaction was stirred at 0℃for 1 hour. After completion of LCMS detection, saturated ammonium chloride solution (2 mL) was added, followed by extraction with dichloromethane (3×20 mL), and the organic phase was dried over anhydrous sodium sulfate, and after removal of the organic solvent by distillation under reduced pressure, isoquinolin-8-ylmethanol 107b (250 mg, yellow oil) was obtained, yield: 74%.
MS m/z(ESI)：160.1[M+1] + 。
Second step
Preparation of 8- (bromomethyl) isoquinoline
Isoquinolin-8-yl methanol 107b (300 mg,1.88 mmol) was dissolved in tetrahydrofuran (5 mL), followed by addition of boron tribromide (763 mg,2.82 mmol) under an ice bath. Stirring was carried out at room temperature 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 concentrating under reduced pressure and removing the solvent, the target product 8- (bromomethyl) isoquinoline 107c (180 mg, yellow oil) was obtained in yield: 34%.
MS m/z(ESI)：222.2[M+1] + 。
Third step
Preparation of 4-fluoro-N- (isoquinolin-8-ylmethyl) -2-methoxy-5-nitroaniline
8- (bromomethyl) isoquinoline 107c (150 mg,0.67 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (188 mg,1.01 mmol) were dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (187 mg,1.35 mmol). The reaction solution 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 a 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%.
MS m/z(ESI)：328.1[M+1] + 。
Fourth step
Preparation of 4-fluoro-6-methoxy-1-N- (quinolin-8-ylmethyl) benzene-1, 3-diamine
Preparation 107d (50 mg,0.15 mmol) 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 the addition of iron powder (85 mg,1.53 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 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, pale yellow oil), yield: 84%.
MS m/z(ESI)：298.1[M+1] + 。
Fifth step
Preparation of dimethyl 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide 111e (40 mg,0.13 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (54 mg,0.16 mmol) and triethylamine (41 mg,0.31 mmol). The reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, compound 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 1107f (80 mg, pale yellow liquid), yield: 66%.
MS m/z(ESI)：539.1[M+1] + 。
Sixth step
Preparation of 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2, 4-dioxo-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 (9 mg,0.22 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (0.1% formic acid/acetonitrile/water) to give 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-039 (20 mg, yellow solid), yield: 54%.
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)
First step
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 (40 mg,0.12 mmol) was dissolved in methanol (10 mL), followed by the addition of acetic acid (0.05 mL), paraformaldehyde (18 mg,0.61 mmol), and the mixture stirred at room temperature for 30 min, followed by the addition of sodium cyanoborohydride (23 mg,0.37 mmol). 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 a 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: 43%.
MS m/z(ESI)：342.1[M+1] + 。
Second step
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 (50 mg,0.15 mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by the addition of iron powder (82 mg,1.46 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 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, pale yellow oil), yield: 79%.
MS m/z(ESI)：312.1[M+1] + 。
Third step
Preparation of dimethyl 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
4-fluoro-2-methoxy-N-methyl-5-nitro-N- (quinolin-8-ylmethyl) aniline 108c (30 mg,0.10 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (39 mg,0.12 mmol) and triethylamine (29 mg,0.29 mmol). The reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, compound 4- [ ({ 2-fluoro-5- [ (isoquinolin-8-ylmethyl) (methyl) amino ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 108d (60 mg, pale yellow liquid), yield: 56%.
MS m/z(ESI)：553.1[M+1] + 。
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 solution in the third step. Lithium hydroxide monohydrate (7 mg,0.16 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (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, pale yellow solid), yield: 21%.
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)
First step
Preparation of quinolin-8-yl methanol
Quinoline 8-carbaldehyde 109a (300 mg,1.9 mmol) was dissolved in methanol (5 mL). Sodium borohydride (42 mg,0.19 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 1 hour. Then dichloromethane (2X 10 mL) was used for extraction, 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
Preparation of 8- (bromomethyl) quinoline
Quinolin-8-yl-methanol 109b (100 mg,0.63 mmol) was dissolved in dichloromethane (5 mL). Triphenylphosphine (399 mg,1.2 mmol) and carbon tetrabromide (416 mg,1.2 mmol) were then added. The reaction mixture was reacted at 25℃for 1 hour. Dichloromethane (2X 5 mL) was then extracted, the organic phases combined, 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] + 。
Third step
Preparation of 4-fluoro-2-methoxy-5-nitro-N- (quinolin-8-methyl) aniline
8- (bromomethyl) quinoline 109c (100 mg,0.47 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (96 mg,0.52 mmol) were dissolved in acetonitrile (10 mL). Potassium carbonate (196 mg,1.52 mmol) and potassium iodide (8 mg, 0.004mmol) were then added, and the reaction mixture was reacted at 25℃for 1 hour. The solvent was then dried by spinning 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- (quinolin-8-methyl) aniline 109d (80 mg, yellow solid) in 46% yield.
MS m/z(ESI)：328.1[M+1] + 。
Fourth step
Preparation of 4-fluoro-6-methoxy-1-N- (quinoline-8-methyl) benzene-1, 3-diamine
4-fluoro-2-methoxy-5-nitro-N- (quinoline-8-methyl) aniline 109d (80 mg,0.24 mmol) 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 was 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- (quinoline-8-methyl) benzene-1, 3-diamine 109e (60 mg, white solid) in 74% yield.
MS m/z(ESI)：287.1[M+1] + 。
Fifth step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (quinolin-8-methyl) amino ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
4-fluoro-6-methoxy-1-N- (quinoline-8-methyl) benzene-1, 3-diamine 109e (30 mg,0.10 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (37 mg,0.11 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (30 mg,0.30 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：539.1[M+1] + 。
Sixth step
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
Dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (quinolin-8-methyl) amino ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 109f (5 mL of stock solution) was dissolved in methanol with lithium hydroxide (3 mg,0.078 mmol): water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=82:18) to give (acetonitrile/0.1% aqueous formic acid) purified 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), yield: 7%.
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)
First step
Preparation of 4-fluoro-2-methoxy-N- (naphthalen-1-ylmethyl) -5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline 110a (300 mg,1.61 mmol), 2- (chloromethyl) naphthalene (313 mg,1.77 mmol), potassium iodide (134 mg,0.81 mmol), potassium carbonate (445 mg,3.22 mmol) 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 the residue obtained after concentration was separated and purified by silica gel column (petroleum ether/ethyl acetate=2/1) to give preparation 110b of 4-fluoro-2-methoxy-N- (naphthalen-1-ylmethyl) -5-nitroaniline (400 mg, orange solid), yield: 61%.
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
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 (200 mg,0.61 mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (686 mg,12.26 mmol) and stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated to give 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] + 。
Third step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (naphthalen-1-ylmethyl) amino ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
Preparation of 4-fluoro-6-methoxy-1-N- (naphthalen-1-ylmethyl) benzene-1, 3-diamine 110c (60 mg,0.20 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (139 mg,1.01 mmol) were dissolved in tetrahydrofuran (5 mL) then triethylamine (113 mg,2.02 mmol) was added and the mixture was stirred at 25℃for 16 hours.
MS m/z(ESI)：538.1[M+1] + 。
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 solution) was dissolved in methanol with lithium hydroxide (16 mg,0.65 mmol): water=3:1 (4 mL). The mixture was reacted at 25℃for 1 hour. After the reaction was completed, it was prepared by direct concentration and preliminary purification by reverse column (water: acetonitrile=3:2), and after purification 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) was obtained in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-043)
First step
Preparation of isoquinoline-8-carboxylic acid
Isoquinoline-8-carboxylic acid methyl ester 111a (300 mg,1.75 mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL), followed by sodium hydroxide (240 mg,4.81 mmol). The reaction was stirred at room temperature for 1 hour. After completion of LCMS detection reaction, water (20 mL) was added after removal of the organic solvent by distillation under reduced pressure, and then isoquinoline-8-carboxylic acid 111b (250 mg, white solid) was obtained by lyophilization, yield: 81%.
MS m/z(ESI)：174.1[M+1] + 。
Second step
Preparation of isoquinoline-8-carbonyl chloride
Preparation 111b (300 mg,1.73 mmol) of isoquinoline-8-carboxylic acid was dissolved in dichloromethane (10 mL) followed by the addition of DMF (0.1 mL) and oxalyl chloride (439 mg,3.46 mmol). Stirring was carried out at room temperature for 1h. After the reaction was completed, the solvent was removed after concentration under reduced pressure to give the objective isoquinoline-8-carbonyl chloride 111c (300 mg, white solid), yield: 81%.
MS m/z(ESI)：188.2(M-Cl+CH 3 )。
Third step
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-carboxamide
4-fluoro-2-methoxy-5-nitroaniline (552 mg,3.13 mmol) was dissolved in ethyl acetate (20 mL), pyridine (247 mg,3.13 mmol) was then added, and isoquinoline-8-carbonyl chloride 111c (300 mg,1.56 mmol) was finally dissolved in ethyl acetate (5 mL) and added to the reaction solution under ice-bath. The reaction solution was stirred at 0℃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 a 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%.
MS m/z(ESI)：342.1[M+1] + 。
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 (50 mg,0.15 mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by the addition of iron powder (82 mg,1.46 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 mL), and 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, pale yellow oil), yield: 79%.
MS m/z(ESI)：312.1[M+1] + 。
Fifth step
Preparation of dimethyl 4- ({ [ 2-fluoro-5- (isoquinolin-8-amino) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-carboxamide 111e (30 mg,0.10 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (39 mg,0.12 mmol) and triethylamine (29 mg,0.29 mmol). The reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, compound 4- ({ [ 2-fluoro-5- (isoquinolin-8-amino) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid dimethyl ester 111f (60 mg, pale yellow liquid), yield: 56%.
MS m/z(ESI)：553.1[M+1] + 。
Sixth step
Preparation of 3- [ 2-fluoro-5- (isoquinoline-8-amino) -4-methoxyphenyl ] -2, 4-dioxo-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 (7 mg,0.16 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (0.1% formic acid/acetonitrile/water) to give 3- [ 2-fluoro-5- (isoquinoline-8-amino) -4-methoxyphenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-043 (5 mg, pale yellow solid), yield: 18%.
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- (phenylsulphonamido) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-044)
First step
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) benzenesulfonamide
4-fluoro-2-methoxy-5-nitroaniline 112a (2000 mg,10.74 mmol) was dissolved in dichloromethane (30 mL), pyridine (850 mg,10.7446 mmol) was added, and stirred at 0deg.C for 5min. Then, benzenesulfonyl chloride (2087 mg,11.82 mmol) was added dropwise to the above reaction solution. 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 solution was extracted with dichloromethane (3×30 mL), and 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
Preparation of tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) (benzenesulfonyl) carbamate
N- (4-fluoro-2-methoxy-5-nitrophenyl) benzenesulfonamide 112b (200 mg,0.61 mmol) was dissolved in dichloromethane (10 mL). Di-tert-butyl dicarbonate (267 mg,1.23 mmol) and triethylamine (186 mg,1.84 mmol) were added sequentially. 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), yield: 65.43%.
MS m/z(ESI)：449.0(M+23)。
Third step
Preparation of tert-butyl N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (phenylsulfonyl) carbamate
Tert-butyl (4-fluoro-2-methoxy-5-nitrophenyl) (benzenesulfonyl) carbamate 112c (150 mg,0.35 mmol) was dissolved in methanol (20 mL) and ammonium chloride (21 mg,0.39 mmol) was added followed by iron powder (196 mg,3.52 mmol). The resulting mixture was stirred at 25℃for 3 hours, and 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, dried over anhydrous sodium sulfate, and concentrated. The resulting residue was further purified with silica gel column (petroleum ether/ethyl acetate=2/1), tert-butyl N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (benzenesulfonyl) carbamate 112d (50 mg, white solid), yield: 32.26%.
MS m/z(ESI)：297.0(M+1-100)。
Fourth step
Preparation of 3- (5- (N- (tert-butoxycarbonyl) phenylsulfamido) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Tert-butyl N- (5-amino-4-fluoro-2-methoxyphenyl) -N- (benzenesulfonyl) carbamate 112d (50 mg,0.14 mmol) and 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (71 mg,0.21 mmol) and triethylamine (144 mg,1.42 mmol) 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 stirred at 25℃for 1h, followed by addition of lithium hydroxide (34 mg,1.42 mmol). The resulting mixture was stirred at 25℃for 1h. After the completion of the reaction, the resulting mixture was washed with 30mL of water. The mixture was then extracted with ethyl acetate (3X 20 mL). The organic phases were collected, combined, dried over anhydrous sodium sulfate and concentrated. The resulting residue was further purified with silica gel column (petroleum ether/ethyl acetate=2/1) to give 112e (60 mg, white solid), yield of 3- (5- (N- (tert-butoxycarbonyl) phenylsulfonamide) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid: 69.49%.
MS m/z(ESI)：492.0(M+1-100)。
Fifth step
Preparation of 3- (2-fluoro-4-methoxy-5- (phenylsulfonamide) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
3- (5- (N- (tert-Butoxycarbonyl) phenylsulfamido) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid 112e (50 mg,0.09 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (104 mg,0.91 mmol) was added. The resulting mixture was stirred at 25℃for 30 minutes, and after completion of the reaction, concentrated. The concentrate was purified by column chromatography (mobile phase: acetonitrile/water=3/1) to give crude product, which was purified by preparative column (mobile phase: 0.1% formic acid/acetonitrile/water) to give 3- (2-fluoro-4-methoxy-5- (phenylsulfanomido) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-044 (21.9 mg, white solid), 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-045)
First step
Preparation of N- (2, 3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (chloromethyl) -3, 4-difluoro-1-methoxybenzene 113a (200 mg,1.0 mmol) was dissolved in acetonitrile (5 mL), followed by addition of 4-fluoro-2-methoxy-5-nitroaniline (3836 mg,2.0 mmol), potassium carbonate (7197 mg,5.2 mmol) and potassium iodide (34 mg,0.2 mmol). The mixture was stirred at 50℃for 2 hours under nitrogen atmosphere. After the completion of the reaction, the reaction mixture 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 113b (80 mg, yellow solid), yield: 20%.
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
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 (80 mg,0.23 mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (130 mg,2.3 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 N1- (2, 3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxybenzene-1, 3-diamine 113c (60 mg, yellow solid), yield: 70%. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.1[M+1] + 。
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 (60 mg,0.19 mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (343 mg,1.0 mmol) and triethylamine (52 mg,0.51 mmol) 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 dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 113d (35 mg, yellow solid), yield: 33%. The crude product was used directly in the next reaction.
MS m/z(ESI)：554.1[M+1] + 。
Fourth step
Preparation of 3- (5- ((2, 3-difluoro-6-methoxybenzyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-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 (35 mg,0.06 mmol) was dissolved in tetrahydrofuran/methanol/water=1: 1:1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-045 (15 mg, yellow solid), yield: 49%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-046)
First step
Preparation of N- (2, 3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (chloromethyl) -3, 4-difluoro-1-methoxybenzene 114a (200 mg,1.0 mmol) was dissolved in acetonitrile (5 mL), followed by addition of 4-fluoro-2-methoxy-5-nitroaniline (3836 mg,2.0 mmol), potassium carbonate (7197 mg,5.2 mmol) and potassium iodide (34 mg,0.2 mmol). The mixture was stirred at 50℃for 2 hours under nitrogen atmosphere. After the completion of the reaction, the reaction mixture 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%.
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
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 (80 mg,0.23 mmol) was dissolved in methanol (5 mL), followed by addition of paraformaldehyde (36 mg,0.58 mmol) and acetic acid (0.35 mg,0.0058 mmol) to the solution. The reaction was stirred at room temperature for 0.5 hours under nitrogen protection. Subsequently, sodium cyanoboroate (18 mg,0.29 mmol) was added to the system, and the reaction solution was stirred at room temperature for 2 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 N- (2, 3-difluoro-6-methoxybenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 114c (60 mg, yellow solid), yield: 70%. The crude product was used directly in the next reaction.
MS m/z(ESI)：357.1[M+1] + 。
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 (60 mg,0.19 mmol) was dissolved in acetonitrile (5 mL), followed by addition of iron powder (130 mg,2.3 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 N1- (2, 3-difluoro-6-methoxybenzyl) -4-fluoro-6-methoxy-N1-toluene-1, 3-diamine 114d (50 mg, yellow solid), yield: 85%. The crude product was used directly in the next reaction.
MS m/z(ESI)：327.1[M+1] + 。
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 (50 mg,0.15 mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (343 mg,1.0 mmol) and triethylamine (52 mg,0.51 mmol) 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 dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 114e (35 mg, yellow solid), yield: 33%. The crude product was used directly in the next reaction.
MS m/z(ESI)：568.1[M+1] + 。
Fifth step
Preparation of 3- (5- ((2, 3-difluoro-6-methoxybenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-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 (35 mg,0.06 mmol) was dissolved in tetrahydrofuran/methanol/water=1: 1:1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-046 (15 mg, yellow solid), yield: 49%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-047)
First step
Preparation of 8- (benzylthio) isoquinoline
8-bromoisoquinoline (1.8 g,8.6 mmol), tris (dibenzylideneacetone) dipalladium (390 mg,0.4 mmol) and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (501 mg,0.9 mmol) were dissolved in 1,4 dioxane (20 mL), and benzyl mercaptan (1.28 g,10.3 mmol) and N, N-diisopropylethylamine (2.22 g,17.2 mmol) were added thereto, and the reaction solution was stirred under nitrogen atmosphere at 110℃for 16h. After completion of the reaction, the reaction mixture was quenched with 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 separated and purified by silica gel column (petroleum ether/ethyl acetate=9/1) to give 8- (benzylthio) isoquinoline 115b (2.1 g, yellow solid) in 95% yield.
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
Preparation of isoquinoline-8-sulfonyl chloride
8- (benzylthio) isoquinoline 115b (1.56 g,6.2 mmol) was dissolved in acetonitrile (20 mL) and acetic acid: water = 2:1 (15 mL) and 1, 3-dichloro-5, 5-dimethylimidazolidine-2, 4-dione (1.47 g,7.4 mmol) were added to the reaction mixture in an ice-water bath, and the reaction mixture was allowed to react at 25℃for 1 hour. After completion of the reaction, the reaction mixture was quenched with 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 separated and purified by silica gel column (petroleum ether/ethyl acetate=7/3) to give isoquinoline-8-sulfonyl chloride 115c (506 mg, yellow solid) in 35% yield.
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)。
Third step
Preparation of N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-sulfonamide
Isoquinoline-8-sulfonyl chloride 115c (500 mg,2.2 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (347 mg,4.9 mmol) were dissolved in dichloromethane (15 mL), pyridine (277 mg,4.4 mmol) was added at room temperature and the reaction was reacted at 25℃for 2 hours. After completion of the reaction, the reaction mixture was quenched with 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 with 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)。
Fourth step
Preparation of N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-sulfonamide
N- (4-fluoro-2-methoxy-5-nitrophenyl) isoquinoline-8-sulfonamide 115d (40 mg,0.11 mmol), iron powder (17 mg,0.31 mmol) and ammonium chloride (17 mg,0.32 mmol) were dissolved in ethanol: water = 1:1 (5 mL), the reaction was allowed to react at 80℃for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (3X 10 mL). All the 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] + 。
Fifth step
Preparation of dimethyl 4- (3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
N- (5-amino-4-fluoro-2-methoxyphenyl) isoquinoline-8-sulfonamide 115e (25 mg,0.07 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (29 mg,0.08 mmol) tetrahydrofuran (3 mL) were added to triethylamine (22 mg,0.2 mmol) at room temperature, the reaction mixture was reacted at room temperature for 16 hours, water (10 mL) was added to quench the reaction mixture after completion of the reaction, extraction was performed with ethyl acetate (3X 10 mL), all organic phases were combined, dried over anhydrous sodium sulfate and concentrated, and the resulting residue was purified using a reverse phase column (54% acetonitrile/water) to give dimethyl 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] + 。
Sixth step
Preparation of 3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 115f (20 mg,0.03 mmol) and lithium hydroxide (3 mg,0.1 mmol) 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, 1M hydrochloric acid (5 mL) was added to the reaction mixture to quench the reaction mixture, and the mixture was extracted with ethyl acetate (3X 20 mL). All organic phases were combined, dried over anhydrous sodium sulfate and concentrated. Purification of the resulting residue by HPLC preparation (mobile phase: acetonitrile/water) gave Cpd-047 (0.72 mg, white solid) as 4% yield of 3- (2-fluoro-5- (isoquinoline-8-sulfonylamino) -4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid.
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)
First step
Preparation of 1-methoxyisoquinoline-8-carboxylic acid methyl ester
Methyl 1-chloroisoquinoline-8-carboxylate 116a (160 mg,0.72 mmol) and sodium methoxide (195 mg,3.61 mmol) were dissolved in methanol (2 mL). The mixture was reacted in a microwave apparatus at 90℃for 4 hours. 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
Preparation of (1-methoxyisoquinolin-8-yl) methanol
1-methoxyisoquinoline-8-carboxylic acid methyl ester 116b (100 mg,0.46 mmol) was dissolved in tetrahydrofuran (5 mL), and DABAL-H/THF solution (2.5 mL) was slowly added dropwise thereto, and the mixture was stirred at 0deg.C for 1 hour. After the reaction was completed, a saturated ammonium chloride solution (5 mL) was added to quench, and extracted with ethyl acetate (3×50 mL), and the organic phases were combined and dried over anhydrous sodium sulfate, and the obtained residue was separated and purified by a silica gel column (petroleum ether/ethyl acetate=2/1) to obtain (1-methoxyisoquinolin-8-yl) methanol 116c (90 mg, white solid), yield: 82%.
MS m/z(ESI)：190.1[M+1] + 。
Third step
Preparation of 8- (chloromethyl) -1-methoxyisoquinoline
(1-methoxyisoquinolin-8-yl) methanol 116c (60 mg,0.31 mmol), tsCl (121 mg,0.63 mmol), DMAP (8 mg,0.06 mmol), triethylamine (64 mg,0.63 mmol) 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 53% yield.
MS m/z(ESI)：208.1[M+1] + 。
Fourth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1-methoxyisoquinoline
8- (chloromethyl) -1-methoxyisoquinoline 116d (40 mg,0.19 mmol), 4-fluoro-2-methoxy-5-nitrophenol (43 mg,0.23 mmol), potassium carbonate (53 mg,0.39 mmol) and potassium iodide (48 mg,0.29 mmol) were dissolved in acetonitrile (5 mL). The mixture was reacted at 80℃for 2 hours. After the reaction was completed, extracted with dichloromethane (3×50 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and the resulting residue was concentrated and purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1-methoxyisoquinoline 116e (50 mg, orange solid), yield: 64%.
MS m/z(ESI)：359.0[M+1] + 。
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 (50 mg,0.14 mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (156 mg,2.79 mmol) and stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated to give 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy group]Aniline 116f (35 mg, brown solid) was produced in 53% yield.
MS m/z(ESI)：329.1[M+1] + 。
Sixth step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] aniline 116f (35 mg,0.11 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (178 mg,0.53 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (59 mg,1.06 mmol) was added and the mixture was stirred at 25℃for 16 hours.
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
Dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (1-methoxyisoquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate 116g (5 mL stock solution) was dissolved in methanol with lithium hydroxide (8 mg,0.35 mmol): water=3:1 (4 mL). The mixture was reacted at 25℃for 1 hour. After the reaction was completed, the mixture was directly concentrated and purified by reverse column (water: acetonitrile=3:2) to give 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, pale yellow solid) in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-049)
First step
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 (180 mg,0.70 mmol) and methylphenylboronic acid (78 mg,1.30 mmol) were dissolved in toluene/ethanol/water (2 mL, 7:2:1). Then, potassium carbonate (181 mg,1.31 mmol) and ferrocene palladium dichloride (151 mg,0.13 mmol) were added, and the reaction solution was subjected to microwave reaction at 80℃for 1 hour under the protection of nitrogen. After completion of the reaction, the solvent was removed under reduced pressure, 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 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
Preparation of 5- (1, 3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyaniline
4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole 120b (50 mg,0.15 mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (84 mg,1.50 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was 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] + 。
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 (47 mg,0.15 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (104 mg,0.31 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (47 mg,0.46 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The reaction solution was not subjected to any post-treatment to obtain a stock solution of dimethyl 4- ({ [5- (1, 3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 120d.
MS m/z(ESI)：546.0[M+1] + 。
Fourth step
Preparation of 3- [5- (1, 3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2, 4-dioxo-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-dicarboxylic acid 121d (5 mL stock solution) and lithium hydroxide (13 mg,0.53 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction solution was reacted at 25℃for an hour. After the reaction was completed, it was directly concentrated and preliminarily purified by reverse column (water: acetonitrile=20:80) to give 3- [5- (1, 3-benzothiazol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-049 (6.45 mg, white solid) in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-050)
First step
Preparation of 2-bromo-4-methyl-1, 3-benzothiazole
4-methyl-1, 3-benzothiazol-2-amine 121a (2.00 g,12.20 mmol) was dissolved in acetonitrile (25 mL). Cuprous bromide (2.10 g,14.60 mmol) and tert-butyl nitrite (1.89 g,0.02 mmol) were added sequentially. The reaction solution was reacted at room temperature for 0.5 hours. 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) in 88% yield.
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
Preparation of 2-bromo-4- (bromomethyl) -1, 3-benzothiazole
2-bromo-4-methyl-1, 3-benzothiazole 121b (500 mg,2.19 mmol) was dissolved in carbon tetrachloride (10 mL) and benzoyl peroxide (106 mg,0.44 mmol) and N-bromosuccinimide (4638 mg,2.63 mmol) were added at room temperature. The reaction solution was reacted at 85℃for 20 hours. After the completion of the reaction, carbon tetrachloride was removed by concentration, and then diluted with methylene chloride (15 mL), washed with water (5 mL) and saturated brine (2×5 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).
Third step
Preparation of 2-bromo-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole
2-bromo-4- (bromomethyl) -1, 3-benzothiazole 121c (300 mg,0.98 mmol) was dissolved in acetonitrile (10 mL), 4-fluoro-2-methoxy-5-nitrophenol (183 mg,0.98 mmol), potassium carbonate (81 mg,0.49 mmol) and potassium iodide (81 mg,0.49 mmol) were added sequentially, and the reaction solution was reacted at 65℃for 1 hour. After completion of the reaction, water (2 mL) was added thereto, 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] + 。
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 (180 mg,0.70 mmol) and methylphenylboronic acid (78 mg,1.30 mmol) were dissolved in toluene/ethanol/water (2 mL, 7:2:1). Then, potassium carbonate (181 mg,1.31 mmol) and ferrocene palladium dichloride (151 mg,0.13 mmol) were added, and the reaction solution was subjected to microwave reaction at 80℃for 1 hour under the protection of nitrogen. After completion of the reaction, the solvent was removed under reduced pressure, 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 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).
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 (40 mg,0.11 mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (64 mg,1.15 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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] + 。
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 (35 mg,0.11 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (74 mg,0.22 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (33 mg,0.33 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The reaction solution 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.
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
121g (5 mL stock solution) of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzothiazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid ester with lithium hydroxide (13 mg,0.53 mmol) was dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) to give 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzothiazol-4-yl) methoxy ] phenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-050 (4.30 mg, white solid) in 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)
First step
Preparation of 1H benzo [ d ] [1,2,3] triazole-4-carboxylic acid
2, 3-diaminobenzoic acid (1000 mg,6.6 mmol) 122a was dissolved in acetic acid (6 mL) and water (9 mL). A solution of sodium nitrite (544 mg,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, the solid was filtered and washed with water to give 1H-1,2, 3-benzotriazole-4-carboxylic acid 122b (1100 mg, grey solid), yield: 92%.
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
Preparation of methyl 1H-1,2, 3-benzotriazole-4-carboxylate
1H-1,2, 3-benzotriazole-4-carboxylic acid 122b (1100 mg,6.6 mmol) was dissolved in methanol (20 mL), and then thionyl chloride (2188 mg,18 mmol) was gradually added dropwise to the solution at 0deg.C, and the reaction solution was stirred at 40deg.C for 16H. 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. Purification of the crude product by silica gel column (MeOH/dcm=10%) afforded 1H-1,2, 3-benzotriazole-4-carboxylic acid methyl ester 122c (906 mg, grey solid), yield: 68%.
MS m/z(ESI)：178.1[M+1] + 。
Third step
Preparation of 1H-1,2, 3-benzotriazol-4-yl methanol
1H-1,2, 3-benzotriazole-4-carboxylic acid methyl ester 122c (900 mg,5.07 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), followed by dropwise addition of diisobutylaluminum hydride solution (1.0M in n-hexane) (10 mL) to the solution at 0 ℃. The mixture was stirred at 0℃for 2 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. Purification of the crude product by silica gel column (MeOH/dcm=10%) afforded 1H-1,2, 3-benzotriazol-4-ylmethanol 122d (540 mg, grey solid), yield: 60%.
MS m/z(ESI)：150.1[M+1] + 。
Fourth step
Preparation of 4- (chloromethyl) -1H benzo [ d ] [1,2,3] triazole
1H-1,2, 3-benzotriazol-4-yl-methanol 122d (540 mg,3.60 mmol) was dissolved in ultra-dry tetrahydrofuran (6 mL) followed by the addition of thionyl chloride (10 mL) at 0deg.C under nitrogen. The reaction was stirred at room temperature for 3 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 122e (363 mg, yellow solid) of 4- (chloromethyl) -1H-benzo [ d ] [1,2,3] triazole, yield: 60%. The crude product was used directly in the next reaction.
MS m/z(ESI)：168.1[M+1] + 。
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 (803 mg,2.16 mmol)), 4-fluoro-2-methoxy-5-nitroaniline (522 mg,2.80 mmol), potassium carbonate (2013 mg,10.8 mmol) and potassium iodide (52 mg,0.21 mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50℃for two hours. After the completion of the reaction, 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. Purification of the crude product by silica gel column (MeOH/dcm=10%) afforded 4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -1H-benzo [ d ] [1,2,3] triazole 122f (300 mg, yellow solid), yield: 44%.
MS m/z(ESI)：319.1[M+1] + 。
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 (300 mg,0.94 mmol) was dissolved in acetonitrile (5 mL) followed by addition of iron powder (650 mg,9.4 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 122g (200 mg, yellow solid) of 5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline, yield: 74%. The crude product was used directly in the next reaction.
MS m/z(ESI)：289.1[M+1] + 。
Seventh step
Preparation of dimethyl 4- (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2, 3-dicarboxylate
122g (200 mg,0.64 mmol) 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 dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (686 mg,2.0 mmol) and triethylamine (162 mg,1.52 mmol) 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 dimethyl 4- (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2, 3-dicarboxylate 122H (85 mg, yellow solid), yield: 27%. 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl- (3- (5- ((1H-benzo [ d ] [1,2,3] triazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) uridine) thiophene-2, 3-dicarboxylic acid 122H (85 mg,0.16 mmol) was dissolved in tetrahydrofuran/methanol/water=1: 1:1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. The crude product was purified by preparation of (FA) to give 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 (8 mg, yellow solid), yield: 10%.
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-benzoxazol-4-yl) methoxy ] phenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-052)
First step
Preparation of 2-methyl-1, 3-benzoxazole-4-carboxylic acid
2-amino-3-hydroxybenzoic acid 123a (2.00 g,0.01 mol) 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 give preparation 123b (1.45 g, orange solid) of 2-methyl-1, 3-benzoxazole-4-carboxylic acid, yield: 63%.
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
Preparation of (2-methyl-1, 3-benzoxazol-4-yl) methanol
Preparation 123b (1.00 g,5.60 mmol) of 2-methyl-1, 3-benzoxazole-4-carboxylic acid was dissolved in tetrahydrofuran (5 mL), and a 1N borane solution in tetrahydrofuran (5 mL) was added dropwise and reacted at 0℃for 3 hours. After the reaction was completed, methanol was added dropwise 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=1/1) to give (2-methyl-1, 3-benzoxazol-4-yl) methanol 123c (356 mg, yellow solid), yield: 39%.
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).
Third step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1, 3-benzoxazole
To a solution of (2-methyl-1, 3-benzoxazol-4-yl) methanol 123c (150 mg,0.92 mmol), 4-fluoro-2-methoxy-5-nitrophenol (206 mg,1.10 mmol) and triphenylphosphine (223 mg,1.10 mmol) in tetrahydrofuran (10 mL) was added dropwise diisopropyl azodicarboxylate (289 mg,1.10 mmol) 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, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=4/1) to give 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methyl-1, 3-benzoxazole 123d (295 mg, white solid), yield: 96%.
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).
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 (284 mg,0.86 mmol) in ethanol (3 mL) and water (3 mL) was added iron powder (144 mg,2.57 mmol) and ammonium chloride (138 mg,2.57 mmol), and the mixture was reacted at 80℃for 1 hour. After completion of the reaction, extracted with ethyl acetate (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzoxazol-4-yl) methoxy ] aniline 123e (246 mg, yellow solid), yield: 94%.
MS m/z(ESI)：303.1[M+1] + 。
Fifth step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
To a solution of 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzoxazol-4-yl) methoxy ] aniline 123e (216 mg,0.71 mmol) and triethylamine (21 mg,2.14 mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (264 mg,0.79 mmol), after the reaction was completed at 25 ℃ for 2 hours, water (5 mL) was added to quench, 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 (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-2, 3-dicarboxylate 123f (154 mg, white solid) in 40% yield.
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).
Sixth step
Preparation of 3- { 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzoxazol-4-yl) methoxy ] phenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
To a solution of 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methyl-1, 3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid dimethyl ester 123f (134 mg,0.25 mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide monohydrate (31 mg,0.74 mmol), and the mixture was reacted at 25℃for 1 hour. After completion of the reaction, 1M diluted hydrochloric acid was added dropwise to adjust ph=6-7, 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 via a preparative column to give Cpd-052 (5 mg, white solid) as 3- { 2-fluoro-5- [ (5-fluoroisoquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid, yield: 4%.
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)
First step
Preparation of 4- (bromomethyl) -1, 3-benzodioxole
1, 3-Benzodioxan-4-ylmethanol 124a (250 mg,1.64 mmol), triphenylphosphine (862mg, 3.29 mmol) and carbon tetrabromide (1.00 g,3.29 mmol) were dissolved in dichloromethane (10 mL). The mixture was stirred at 25 ℃ for 2 hours and concentrated, and the resulting residue was purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 4- (bromomethyl) -1, 3-benzodioxole 124b (220 mg, white oil), yield: 50%.
1 H NMR(400MHz,CDCl 3 )δ6.82-6.78(m,3H),6.02(s,2H),4.47(s,2H).
Second step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzodioxole
4- (bromomethyl) -1, 3-benzodioxole 124b (120 mg,0.56 mmol), 4-fluoro-2-methoxy-5-nitrophenol (104 mg,0.56 mmol), potassium carbonate (154 mg,1.11 mmol) and potassium iodide (139 mg,0.84 mmol) 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 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), yield: 35%.
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).
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 (80 mg,0.25 mmol) was dissolved in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (279 mg,4.98 mmol) and the mixture was stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated to give preparation 124d (50 mg, brown oil) of 5- (1, 3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyaniline. The yield was 55%.
MS m/z(ESI)：292.1[M+1] + 。
Fourth step
Preparation of dimethyl 4- ({ [5- (1, 3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
Preparation 124d (50 mg,0.17 mmol) of 5- (1, 3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyaniline and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (288 mg,0.86 mmol) were dissolved in tetrahydrofuran (5 mL), triethylamine (90 mg,1.72 mmol) was then added and the mixture was stirred at 25℃for 16 hours.
MS m/z(ESI)：533.0[M+1] + 。
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
Dimethyl 4- ({ [5- (1, 3-benzodioxol-4-ylmethoxy) -2-fluoro-4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 124e (5 mL stock solution) was dissolved in methanol with lithium hydroxide (27 mg,1.13 mmol): water=3:1 (4 mL). The mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was prepared by directly concentrating and preliminary purifying with a reverse column (water: acetonitrile=3:2), and after purifying, 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) was obtained in 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)
First step
Preparation of 1H-indazol-7-yl methanol
1H-indazole-7-carboxylic acid methyl ester 125a (50 mg,0.28 mmol) was dissolved in tetrahydrofuran (5 mL). Diisobutylaluminum hydride (5 mL) was then added at 0deg.C. The reaction mixture was reacted at 25℃for 1 hour. Then quenched with saturated ammonium chloride solution, extracted with dichloromethane (2X 10 mL), the organic phases combined and dried over anhydrous sodium sulfate, and the concentrated residue purified by column chromatography (petroleum ether/ethyl acetate: 50/50) to give 1H-indazol-7-ylmethanol 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
Preparation of 7- (bromomethyl) -1H-indazole
1H-indazol-7-yl-methanol 125b (120 mg,0.81 mmol) was dissolved in dichloromethane (5 mL). Triphenylphosphine (424 mg,1.62 mmol) and carbon tetrabromide (537 mg,1.62 mmol) were then added. The reaction mixture was reacted at 25℃for 1 hour. Dichloromethane (2X 5 mL) was then extracted, the organic phases combined, 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) in 52% yield.
MS m/z(ESI)：211.0[M+1] + 。
Third step
Preparation of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1H-indazole
7- (bromomethyl) -1H-indazole 125c (100 mg,0.47 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (97 mg,0.52 mmol) were dissolved in acetonitrile (10 mL). Potassium carbonate (196 mg,0.28 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The solvent was then dried by spinning and extracted with dichloromethane (2X 10 mL). The combined organic phases were 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) in 85% yield.
MS m/z(ESI)：318.1[M+1] + 。
Fourth step
Preparation of 2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyaniline
7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1H-indazole 125d (140 mg,0.57 mmol) 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 was 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] + 。
Fifth step
Preparation of dimethyl 4- ({ [ [ 2-fluoro-5- (1H-indazol-7-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate
2-fluoro-5- (1H-indazole-7-methoxy) -4-methoxyaniline 125e (30 mg,0.10 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (42 mg,0.12 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (31 mg,0.31 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：529.1[M+1] + 。
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-dicarboxylic acid 125f (5 mL stock solution) and lithium hydroxide (3 mg,0.078 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=82:18) to give 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) in a yield of 13.91%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-055)
First step
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 (40 mg,0.12 mmol) was dissolved in jiachun (10 mL), acetic acid (0.05 mL), paraformaldehyde (18 mg,0.61 mmol) was then added, the mixture stirred at room temperature for 30 min, and finally sodium cyanoborohydride (23 mg,0.37 mmol) was added. 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 a 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%.
MS m/z(ESI)：360.2[M+1] + 。
Second step
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 (40 mg,0.11 mmol) was dissolved in methanol (5 mL) and saturated ammonium chloride (1 mL), followed by the addition of iron powder (62 mg,1.11 mmol). The reaction was stirred at 25℃for 2h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 mL), and 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, pale yellow oil), yield: 65%.
MS m/z(ESI)：330.1[M+1] + 。
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 (25 mg,0.08 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (31 mg,0.09 mmol) and triethylamine (23 mg,0.23 mmol). The reaction solution was stirred at 25℃for 16 hours. After completion of LCMS monitoring the reaction, 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, pale yellow liquid), yield: 58%.
MS m/z(ESI)：571.2[M+1] + 。
Fourth step
3- (2-fluoro-4-methoxy-5- (methyl ((2-methyl-1H-isoquinolin-8-yl) methyl) amino) phenyl) -2, 4-dioxo-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 (7 mg,0.16 mmol) was added. The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-055 (5 mg, pale yellow solid), yield: 18%.
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)
First step
Preparation of [ (2-bromo-3-fluorophenyl) methyl ] (2, 2-dimethoxyethyl) amine
2-bromo-3-fluorobenzaldehyde 127a (4.5 g,22.2 mmol) was dissolved in ethanol (60 mL). 2, 2-Dimethoxyethylamine (2.57 g,24.4 mmol) was then added at 25 ℃. The reaction mixture was reacted at 80℃for 16 hours. The solvent was then dried by spin-drying, extracted with ethyl acetate (2X 50 mL), the organic phases were combined, 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
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 (4.2 g,1.86 mmol) was dissolved in dichloromethane (60 mL). Triethylamine (4.36 g,0.04 mmol) and 4-toluenesulfonyl chloride (8.24 g,0.04 mmol) were then added at 0deg.C. The reaction solution was reacted at 0℃for 1 hour. Then water (2X 50 mL), the organic phases were combined, 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).
Third step
Preparation of 8-bromo-7-fluoroisoquinoline
N- [ (2-bromo-3-fluorophenyl) methyl ] -N- (2, 2-dimethoxyethyl) -4-methyl-1-sulfonylamine 127c (3.6 g,0.008 mmol) was dissolved in dichloromethane (10 mL) and added dropwise to a solution of anhydrous aluminum trichloride (6.4 g,0.048 mmol) in dichloromethane (5 mL) at 0deg.C. The reaction mixture was reacted at 25℃for 16 hours. Then quenched with ice 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] + 。
Fourth step
Preparation of methyl 7-fluoroisoquinoline-8-carboxylate
8-bromo-7-fluoroisoquinoline 127d (1.2 g,5 mmol) was dissolved in methanol (12 mL). Triethylamine (1.55 g,15 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride) (370 mg,0.0 mmol) were then added, and the reaction mixture was pressurized and sealed using a carbon monoxide-filled canister and 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).
Fifth step
Preparation of (7-fluoroisoquinolin-8-yl) methanol
7-fluoroisoquinoline-8-carboxylic acid methyl ester 127e (200 mg,0.97 mmol) was dissolved in tetrahydrofuran (5 mL). Lithium aluminum hydride (184 mg,4.87 mmol) was then added at 0deg.C, and the reaction solution was reacted at 0deg.C for 0.5 hours. Ice water was then added to quench and extracted with dichloromethane (2×5 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] + 。
Sixth step
Preparation of 8- (bromomethyl) -7-fluoroisoquinoline
(7-fluoroisoquinolin-8-yl) methanol 127f (70 mg,0.35 mmol) and triethylamine (205 mg,2.03 mmol) were dissolved in dichloromethane (5 mL). Then, p-toluenesulfonyl chloride (3836 mg,2.03 mmol) and 4-dimethylaminopyridine (8 mg,0.068 mmol) were added, 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 (100 mg,0.51 mmol) of 8- (bromomethyl) -7-fluoroisoquinoline and 4-fluoro-2-methoxy-5-nitrophenol (105 mg,0.56 mmol) were dissolved in acetonitrile (10 mL). Potassium carbonate (211 mg,1.5 mmol) was then added and the reaction was allowed to react at 25℃for 2 hours. The solvent was then dried by spinning 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 for 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 (60 mg,0.17 mmol) 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 was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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] + 。
Ninth step
Preparation of dimethyl 4- [ ({ [5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
5- [ (7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 127i (30 mg,0.09 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (31 mg,0.09 mmol) were dissolved in tetrahydrofuran (5 mL.) then triethylamine (28 mg,0.28 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：558.1[M+1] + 。
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-dicarboxylic acid 127j (5 mL stock solution) and lithium hydroxide (3 mg,0.078 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=82:18) to give 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) in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-057)
First step
Preparation of (2-bromo-3, 4-difluorophenyl) methanol
2-bromo-3, 4-difluorobenzoic acid 129a (5000 mg,21.1 mmol) was dissolved in ultra-dry tetrahydrofuran (50 mL), followed by dropwise addition of borane tetrahydrofuran solution (1M) (10 mL) under nitrogen at 0deg.C. 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 crude product obtained was purified by silica gel column (ethyl acetate/petroleum ether=20%) to give (2-bromo-3, 4-difluorophenyl) methanol 129b (4600 mg, white solid), yield: 88%.
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
Preparation of 2-bromo-3, 4-difluorobenzaldehyde
(2-bromo-3, 4-difluorophenyl) methanol 129b (4600 mg,20.7 mmol) was dissolved in tetrahydrofuran (20 mL), and then manganese dioxide (3898 mg,44.9 mmol) was gradually added to the solution, and the reaction solution was stirred at room temperature for 4 hours. After the reaction was completed, the solid waste was filtered off, the filtrate was 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-3, 4-difluorobenzaldehyde 129c (4000 mg, grey solid), yield: 86%.
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).
Third step
Preparation of N- (2-bromo-3, 4-difluorobenzyl) -2, 2-dimethoxyethane-1-amine
2-bromo-3, 4-difluorobenzaldehyde 129c (4000 mg,18.1 mmol) was dissolved in ethanol solution (30 mL) followed by the addition of 2, 2-dimethoxyethane-1-amine (2093 mg,19.9 mmol). The reaction solution was stirred at 80℃for 16 hours under nitrogen atmosphere. After the reaction was completed, after the reaction solution was cooled to room temperature, sodium cyanoborocyanide (9099 mg,144.80 mmol) was added, the reaction solution was stirred at room temperature for 1 hour, then the mixture was concentrated, 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 by 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%.
MS m/z(ESI)：310.0[M+1] + 。
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 (3000 mg,9.7 mmol) was dissolved in e-dichloromethane solution (20 mL) followed by the addition of triethylamine (4078 mg,40 mmol) and 4-toluenesulfonyl chloride (4610 mg,24 mmol) at 0deg.C. The reaction solution was stirred at room temperature for 3 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. The crude product was purified by 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%.
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).
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 the solution was then slowly added to a mixture of aluminum trichloride (10026 mg,75.2 mmol) in dichloromethane (20 mL) under nitrogen at 0deg.C. The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, 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. Purification of the crude product by silica gel column (dichloromethane/petroleum ether=80%) gave 8-bromo-6, 7-difluoroisoquinoline 129f (1062 mg, orange solid), yield: 52%.
MS m/z(ESI)：244.0[M+1] + 。
Sixth step
Preparation of methyl 6, 7-difluoroisoquinoline-8-carboxylate
8-bromo-6, 7-difluoroisoquinoline 129f (1062 mg,4.4 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (750 mg,1.10 mmol) and triethylamine (896 mg,8.9 mmol) were dissolved in methanol (20 mL), followed by stirring the reaction solution at 100℃under carbon monoxide (1.5 MPa) 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=20%) afforded 129g (600 mg, grey solid) of 6, 7-difluoroisoquinoline-8-carboxylic acid methyl ester, yield: 61%.
MS m/z(ESI)：224.1[M+1] + 。
Seventh step
Preparation of (6, 7-difluoroisoquinolin-8-yl) methanol
129g (600 mg,2.7 mmol) of methyl 6, 7-difluoroisoquinoline-8-carboxylate was dissolved in anhydrous tetrahydrofuran (10 mL) followed by dropwise hydrogenation of a lithium aluminum solution (1.0M in tetrahydrofuran) (3 mL) at 0deg.C. The mixture was stirred at room temperature for 1 hour. 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 (6, 7-difluoroisoquinolin-8-yl) methanol 129h (300 mg, gray solid), yield: 56%.
MS m/z(ESI)：196.1[M+1] + 。
Eighth step
Preparation of 8- (bromomethyl) -6, 7-difluoroisoquinoline
(6, 7-difluoroisoquinolin-8-yl) methanol (300 mg,1.53 mmol) was dissolved in dichloromethane (5 mL), and phosphorus tribromide (0.5 mL) was added dropwise to the solution at 0deg.C. 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) -6, 7-difluoroisoquinoline 129i (180 mg, yellow solid), yield: 46%.
MS m/z(ESI)：258.0[M+1] + 。
Ninth step
Preparation of 6, 7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) isoquinoline
8- (bromomethyl) -6, 7-difluoroisoquinoline 129i (100 mg,0.39 mmol), 4-fluoro-2-methoxy-5-nitroaniline (250 mg,1.40 mmol), potassium carbonate (1011 mg,5.4 mmol) and potassium iodide (52 mg,0.21 mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50℃for 2 hours. After the completion of the reaction, 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=50%) afforded 6, 7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) isoquinoline 129j (55 mg, yellow solid), yield: 40%.
MS m/z(ESI)：365.1[M+1] + 。
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 (55 mg,0.15 mmol) was dissolved in acetonitrile (5 mL) followed by addition of iron powder (650 mg,9.4 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 (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%. The crude product was used directly in the next reaction.
MS m/z(ESI)：335.1[M+1] + 。
Eleventh step
Preparation of dimethyl 4- (3- (5- ((6, 7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
5- ((6, 7-Difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline 129k (50 mg,0.13 mmol) was dissolved in tetrahydrofuran (6 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (686 mg,2.0 mmol) and triethylamine (162 mg,1.52 mmol) 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 129l (65 mg, yellow solid) of dimethyl 4- (3- (5- ((6, 7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid, yield: 82%. 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Preparation method
Dimethyl 4- (3- (5- ((6, 7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 129l (65 mg,0.11 mmol) was dissolved in tetrahydrofuran/water=1: 1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. The crude product was purified by preparation of (FA) to give 3- (5- ((6, 7-difluoroisoquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-057 (12 mg, yellow solid), yield: 20%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-058)
First step
Preparation of 7-fluoro-8-methylquinoline
3-fluoro-2-methylaniline 130a (2.50 g,20.00 mmol) and sodium iodide (450 mg,3.00 mmol) were dissolved in concentrated sulfuric acid (6.8 mL). After stirring the flask at 140℃for 0.5 h, glycerol (2.12 g,20.00 mmol) was slowly added. The reaction solution was reacted at 140℃for 2 hours. After the completion of the reaction, the reaction mixture was returned to room temperature, and the reaction mixture was poured into ice water, ph=8-1 was adjusted with a saturated sodium carbonate solution, and then extracted with ethyl acetate (3×100 mL). Then, the organic phase was combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate=90/10) to give 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
Preparation of 8- (bromomethyl) -7-fluoroquinoline
7-fluoro-8-methylquinoline 130b (500 mg,3.10 mmol) was dissolved in carbon tetrachloride (10 mL), and benzoyl peroxide (150 mg,3.10 mmol) and N-bromosuccinimide (292 mg,3.72 mmol) were added at room temperature. The reaction solution was reacted at 85℃for 4 hours. After the reaction was completed, carbon tetrachloride was removed by concentration, and then diluted with methylene chloride (15 mL), washed with water (5 mL) and saturated brine (2×5 mL) in this order, the organic phases were combined, 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).
Third step
Preparation of 7-fluoro-8- (5-fluoro-2-methoxy-4-nitrophenoxymethyl) quinoline
8- (bromomethyl) -7-fluoroquinoline 130c (100 mg,0.42 mmol) was dissolved in acetonitrile (5 mL), 4-fluoro-2-methoxy-5-nitrophenol (94 mg,0.50 mmol), potassium carbonate (174 mg,1.26 mmol) and potassium iodide (28 mg,0.17 mmol) were sequentially added, and the reaction solution was reacted at 65℃for 1 hour. After completion of the reaction, water (2 mL) was added thereto, 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] + 。
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 (85 mg,0.25 mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (137 mg,2.46 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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] + 。
Fifth step
Preparation of dimethyl 4- [ ({ 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyaniline 130e (80 mg,0.25 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (158 mg,0.50 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (77 mg,0.75 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The reaction solution was not subjected to any post-treatment to obtain dimethyl 4- [ ({ 2-fluoro-4- [ (7-fluoroquinolin-8-yl) methoxy ] -5-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate 130f.
MS m/z(ESI)：558.2[M+1] + 。
Sixth step
Preparation of 3- { 2-fluoro-5- [ (7-fluoroquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2, 4-dioxo-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-dicarboxylic acid 130f (5 mL of stock solution) and lithium hydroxide (13 mg,0.53 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction solution was reacted at room temperature for 2 hours. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) to give 3- { 2-fluoro-5- [ (7-fluoroquinolin-8-yl) methoxy ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-058 (75.60 mg, yellow solid) in a yield of 97.00%.
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)
First step
Preparation of 1, 2-difluoro-3-methyl-4-nitrobenzene
HNO of 3 (65%, 6.80g,0.07 mol) in H 2 SO 4( 15 mL), the mixture was stirred at 0deg.C for 10min, and 1, 2-difluoro-3-methylbenzene 131a (6.00 g,0.05 mol) was slowly added thereto, followed by stirring for 10min. After the reaction was completed, the reaction solution was quenched in ice water, extracted with ethyl acetate (3×50 mL), the organic phases were combined and dried over anhydrous sodium sulfate, and 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) in yield: 47%.
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
Preparation of 3, 4-difluoro-2-methylaniline
1, 2-difluoro-3-methyl-4-nitrobenzene 131b (4.80 g,0.0277 mol) was dissolved in NH 4 To a solution of Cl/MeCN (20 mL), iron powder (31.02 g,0.5539 mol) was added and stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated, and the residue was separated and purified by silica gel column (petroleum ether/ethyl acetate=2/1) to give 3, 4-difluoro-2-methylaniline 131c (3 g, brown oil) in yield: 56%.
MS m/z(ESI)：144.1[M+1] + 。
Third step
Preparation of 6, 7-difluoro-8-methylquinoline
3, 4-difluoro-2-methylaniline 131c (1.40 g,0.0098 mol) and NaI (0.73 g,0.0049 mol) were dissolved in H 2 SO 4( 15 mL), slowly adding glycerol (5.42 g,0.0588 mol) after the temperature of the oil bath Wen rises to 140 ℃, stirring for 3 hours, pouring the reaction solution into glacial methanol for quenching after the reaction is finished, filtering, adjusting the pH to be=9 after the filtrate is dried by spinning, extracting by ethyl acetate (3×50 mL), drying the organic phase by anhydrous sodium sulfate after the combination, concentrating, separating and purifying the obtained residue by a silica gel column (petroleum ether/ethyl acetate=6/1) to obtain 6, 7-difluoro-8-methylquinoline 131d (120 mg)Pale yellow solid), yield: 6%.
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).
Fourth step
Preparation of 8- (bromomethyl) -6, 7-difluoroquinoline
6, 7-difluoro-8-methylquinoline 131d (110 mg,0.61 mmol), BPO (74 mg,0.31 mmol) and NBS (153 mg,0.86 mmol) were dissolved in CCl 4( 6 mL) of the solution, the mixture was stirred under nitrogen at 85℃for 5 hours. After the reaction was completed, the mixture was concentrated, and the residue was separated and purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 8- (bromomethyl) -6, 7-difluoroquinoline 131e (106 mg, white solid), yield: 60%.
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).
Fifth step
Preparation of 6, 7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline
8- (bromomethyl) -6, 7-difluoroquinoline 131e (86 mg,0.33 mmol), 4-fluoro-2-methoxy-5-nitrophenol (75 mg,0.40 mmol), potassium carbonate (92 mg,0.67 mmol) and potassium iodide (83 mg,0.50 mmol) were dissolved in acetonitrile (10 mL). The mixture was reacted at 80℃for 2 hours. After the reaction was completed, extracted with dichloromethane (3×50 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and the resulting residue was concentrated and purified by silica gel column (petroleum ether/ethyl acetate=3/1) to give 6, 7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 131f (100 mg, yellow solid), yield: 65%.
MS m/z(ESI)：365.0[M+1] + 。
Sixth step
Preparation of 5- [ (6, 7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline
The 6, 7-difluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinolineThe solution of the line 131f (80 mg,0.2196 mmol) in NH 4 To a solution of Cl/MeCN (4 mL) was added iron powder (246 mg,4.39 mmol) and the mixture was stirred at 25℃for 1 hour. The reaction solution was filtered and concentrated to give 5- [ (6, 7-difluoroquinolin-8-yl) methoxy group]-2-fluoro-4-methoxyaniline 131g (80 mg, brown solid), yield 76%.
MS m/z(ESI)：335.1[M+1] + 。
Seventh step
Preparation of dimethyl 4- [ ({ 5- [ (6, 7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
Preparation method
131g (70 mg,0.21 mmol) of 5- [ (6, 7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (351 mg,1.05 mmol) were dissolved in tetrahydrofuran (10 mL), triethylamine (117 mg,2.10 mmol) was then 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
Dimethyl 4- [ ({ 5- [ (6, 7-difluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 131h (10 mL of stock solution) with lithium hydroxide (50 mg,2.10 mmol) was dissolved in methanol: water=3:1 (4 mL). The mixture was reacted at 25℃for 1 hour. After the reaction was completed, it was prepared by direct concentration and preliminary purification by reverse column (water: acetonitrile=3:2), and after purification, 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) was obtained in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-060)
First step
Preparation of 1, 5-difluoro-2-methyl-3-nitrobenzene
2, 4-Difluorotoluene (1.00 g,7.80 mmol) and nitric acid (740 mg,11.70 mmol) were dissolved in sulfuric acid (4 mL), and the reaction solution was stirred at 0deg.C for 1h. After the reaction, sodium hydrogencarbonate was added to the reaction mixture to quench the mixture to pH >7, and the mixture was 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
Preparation of 3, 5-difluoro-2-methylaniline
1, 5-difluoro-2-methyl-3-nitrobenzene 132b (600 mg,3.42 mmol), iron powder (580 mg,10.40 mmol) and ammonium chloride (554 mg,10.40 mmol) 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 with 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 separated and purified by silica gel column (petroleum ether/ethyl acetate=9/1) to give 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)。
Third step
Preparation of 5, 7-difluoro-8-methylquinoline
3, 5-difluoro-2-methylaniline 132c (400 mg,2.80 mmol) and sodium iodide (47 mg,0.28 mmol) were dissolved in sulfuric acid (8 mL), glycerin (326 mg,3.10 mmol) 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, and the mixture was filtered and concentrated. Subsequently, a saturated sodium bicarbonate solution was 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 with 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)。
Fourth step
Preparation of 8- (bromomethyl) -5, 7-difluoroquinoline
5, 7-difluoro-8-methylquinoline 132d (96 mg,0.53 mmol), N-bromosuccinimide (114 mg,0.64 mmol) and benzoyl peroxide (65 mg,0.27 mmol) were dissolved in carbon tetrachloride (4 mL), and the reaction solution was reacted at 85℃for 6 hours under a nitrogen atmosphere. After completion of the reaction, the reaction mixture was quenched with 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 with 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)。
Fifth step
Preparation of 5, 7-difluoro-8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoline
8- (bromomethyl) -5, 7-difluoroquinoline 132e (66 mg,0.26 mmol), 4-fluoro-2-methoxy-5-nitrophenol (57 mg,0.31 mmol) and potassium carbonate (42 mg,0.31 mmol) 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 with 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)。
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 (59 mg,0.12 mmol), iron powder (45 mg,0.81 mmol) and ammonium chloride (43 mg,0.81 mmol) were dissolved in ethanol: water = 1:1 (3 mL), the reaction was allowed to react at 60℃for 3 hours. After completion of the reaction, the reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (3X 20 mL). All organics were combined, dried over anhydrous sodium sulfate and concentrated to give 132g (55 mg, grey solid) of 5- ((5, 7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline 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
132g (55 mg,0.12 mmol) of 5- ((5, 7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyaniline and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (83 mg,0.25 mmol) were dissolved in tetrahydrofuran (10 mL), triethylamine (33 mg,0.33 mmol) was added to the reaction at room temperature, the reaction mixture was quenched with water (20 mL) after the reaction was completed at 25 ℃ for 16 hours, extracted with ethyl acetate (3×20 mL), all the organic phases were combined, dried over anhydrous sodium sulfate and concentrated, and the resulting residue was isolated and purified with a silica gel column (petroleum ether/ethyl acetate=3/2) to give dimethyl 4- (3- (5- (((5, 7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 132h (59 mg, yellow solid) in 58% yield.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- (((5, 7-difluoroquinolin-8-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 132H (45 mg,0.08 mmol) and lithium hydroxide (18 mg,0.78 mmol) were dissolved in tetrahydrofuran: methanol: water=1:1:1 (4.5 mL), the reaction was reacted at 25 ℃ for 1 hour.
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)
First step
1, 5-difluoro-2-methyl-3-nitrobenzene 148a (5 g,39 mmol) was dissolved in concentrated sulfuric acid (20 mL). Nitric acid (2.70 g,42.9 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 1 hour. Then ice quenched and the pH was adjusted to 7-8 with 1M sodium hydroxide solution. Extraction with methylene chloride and purification of the concentrated residue by column chromatography (petroleum 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
Preparation of 3, 5-difluoro-2-methylaniline
1, 5-difluoro-2-methyl-3-nitrobenzene 134b (500 mg,2.88 mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (241 mg,4.33 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was 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] + 。
Third step
N- (3, 5-difluoro-2-methylphenyl) acetamide
3, 5-difluoro-2-methylaniline 134c (4.5 g,31.4 mmol) was dissolved in dichloromethane (50 mL) and then acetyl chloride (7.39 g,94.1 mmol) and triethylamine (9.51 g,94.1 mmol) were added dropwise at 25 ℃. The reaction mixture was reacted at 25℃for 1 hour. The solvent was then dried by spinning, extracted with aqueous sodium bicarbonate 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] + 。
Fourth step
N- (3, 5-difluoro-2-methyl-6-nitrophenyl) acetamide
N- (3, 5-difluoro-2-methylphenyl) acetamide 134d (5 g,27.0 mmol) was dissolved in concentrated sulfuric acid (10 mL). Nitric acid (1.87 g,29.7 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 1 hour. Then ice quenched and the pH was adjusted to 7-8 with 1M sodium hydroxide solution. Extraction with methylene chloride 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] + 。
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 solution was reacted at 90℃for 4 hours. Then spin-dry the solvent and quench with ice, adjust pH to 7-8 with 1M sodium hydroxide solution. Extraction with methylene chloride 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] + 。
Sixth step
Preparation of 4, 6-difluoro-3-methylbenzene-1, 2-diamine
3, 5-difluoro-2-methyl-6-nitroaniline 134f (2.1 g,11.2 mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (0.63 g,11.2 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give 134g (1.5 g, red solid) of 4, 6-difluoro-3-methylbenzene-1, 2-diamine in 76% yield.
MS m/z(ESI)：159.0[M+1] + 。
Seventh step
Preparation of 6, 8-difluoro-5-methylquinoxaline
134g (1.5 g,9.5 mmol) of 4, 6-difluoro-3-methylbenzene-1, 2-diamine was dissolved in tetrahydrofuran (9 mL). Then, hydrogen fluoride (4.5 mL) was added thereto, 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 (200 mg,1.11 mmol) was dissolved in carbon tetrachloride (15 mL). Dibenzoyl peroxide (134 mg,0.55 mmol) and N-bromosuccinimide (237 mg,1.33 mmol) were then added at 25℃and the reaction was allowed to react 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] + 。
Ninth step
Preparation of 6, 8-difluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline
134i (300 mg,1.16 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (216 mg,1.16 mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (320 mg,2.32 mmol) and potassium iodide (288 mg,1.73 mmol) were added, and the reaction mixture was reacted at 65℃for 2 hours. The solvent was then dried by spinning 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] + 。
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 (150 mg,0.4 mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (22 mg,0.41 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and 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] + 。
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 (60 mg,0.18 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (60 mg,0.18 mmol) were dissolved in tetrahydrofuran (10 mL.) then triethylamine (54 mg,0.53 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：577.0[M+1] + 。
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
Dimethyl 4- [ ({ 5- [ (6, 8-difluoroquinoxalin-5-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 148l (10 mL stock solution) and lithium hydroxide (5 mg,0.13 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by a reverse column (water: acetonitrile=5:95) to give 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), yield: 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)
First step
Preparation of 4-methyl-3H-1, 3-benzoxazole-2-thione
2-amino-3-methylphenol 137a (3.80 g,30.86 mmol) was dissolved in anhydrous methanol (50 mL) and then potassium ethyl sulfamethacetoacetate (9.96 g,60.71 mmol) was added. The reaction was stirred at 90℃for 4 hours. After LCMS detection reaction was completed, the reaction was cooled to room temperature. After removal of the solvent by distillation under reduced pressure, purification by silica gel column (petroleum ether/ethyl acetate=1/1) gave the product 4-methyl-3H-1, 3-benzoxazole-2-thione 137b (5.0 g, white solid), yield: 88%.
MS m/z(ESI)：166.2[M+1] + 。
Second step
Preparation of 2-chloro-4-methyl-1, 3-benzoxazole
4-methyl-3H-1, 3-benzoxazole-2-thione 137b (2.00 g,12.11 mmol) was dissolved in dichloromethane (10 mL) and phosphorus oxychloride (10 mL) followed by the addition of phosphorus pentachloride (2.77 g,13.32 mmol). The reaction was stirred at 100℃for 16 hours. After LCMS detection reaction was completed, the reaction was cooled to room temperature. After the solvent was distilled off under reduced pressure, ice was added thereto, 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 and distilling off the solvent under reduced pressure, the product 2-chloro-4-methyl-1, 3-benzoxazole 137b (500 mg, pale yellow solid) was obtained after purification by silica gel column (petroleum ether/ethyl acetate=1/1), yield: 19%.
MS m/z(ESI)：168.2[M+1] + 。
Third step
Preparation of 2-methoxy-4-methyl-1, 3-benzoxazole
4-methyl-3H-1, 3-benzoxazole-2-thione 137c (220 mg,1.31 mmol) was dissolved in methanol (10 mL), followed by sodium methoxide (142 mg,2.63 mmol). The reaction was refluxed at 90 ℃ for 5 hours. After LCMS monitored the reaction was complete, the reaction was cooled to room temperature and the solvent was distilled off under reduced pressure. Purification over a silica gel column (petroleum ether/ethyl acetate=1/1) gave the product 2-methoxy-4-methyl-1, 3-benzoxazole 137d (180 mg, white solid), yield: 76%.
MS m/z(ESI)：164.2[M+1] + 。
Fourth step
Preparation of 4- (bromomethyl) -2-methoxy-1, 3-benzoxazole
Isoquinolin-8-yl methanol 137d (200 mg,1.23 mmol) was dissolved in carbon tetrachloride (5 mL), followed by the addition of N-bromosuccinimide (305 mg,1.72 mmol) and benzoyl peroxide (148 mg,0.61 mmol). The reaction was stirred at 80℃for 4h. After the completion of the reaction, the solvent was distilled off under reduced pressure and purified by a silica gel column to give 4- (bromomethyl) -2-methoxy-1, 3-benzoxazole 137e (130 mg, pale yellow solid) as a product, yield: 39%.
MS m/z(ESI)：242.0[M+1] + 。
Fifth step
Preparation of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1, 3-benzoxazole
4- (bromomethyl) -2-methoxy-1, 3-benzoxazole 137e (100 mg,0.41 mmol) and 4-fluoro-2-methoxy-5-nitroaniline (77 mg,0.41 mmol) were dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (114 mg,0.83 mmol). The reaction solution was stirred at 50℃for 4h. 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 a silica gel column (petroleum ether/ethyl acetate=4/1) to give preparation 137f (120 mg, yellow solid), yield of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1, 3-benzoxazole: 67%.
MS m/z(ESI)：349.2[M+1] + 。
Sixth step
Preparation of 2-fluoro-4-methoxy-5- [ (2-methoxy-1, 3-benzoxazol-4-yl) methoxy ] aniline
Preparation 137f (100 mg,0.29 mmol) of 4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2-methoxy-1, 3-benzoxazole was dissolved in acetonitrile (10 mL) and saturated ammonium chloride (2 mL), followed by the addition of iron powder (160 mg,2.87 mmol). The reaction was stirred at 25℃for 0.5h. After completion of the reaction, the iron powder was filtered off, then extracted with dichloromethane (3×20 mL), and 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%.
MS m/z(ESI)：319.1[M+1] + 。
Seventh step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1, 3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2, 3-dicarboxylate
137g (40 mg,0.13 mmol) of 2-fluoro-4-methoxy-5- [ (2-methoxy-1, 3-benzoxazol-4-yl) methoxy ] aniline is dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (51 mg,0.15 mmol) and triethylamine (38 mg,0.38 mmol). The reaction solution was stirred at 25℃for 16 hours. After LCMS monitoring the completion of the reaction, the solvent was distilled off under reduced pressure and purified by silica gel column to give compound 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1, 3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2, 3-dicarboxylic acid dimethyl ester prepared 137h (60 mg, yellow solid), yield: 68%.
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 dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (2-methoxy-1, 3-benzoxazol-4-yl) methoxy ] phenyl } carbamoyl) thiophene-2, 3-dicarboxylate 137h (60 mg,0.07 mmol) was dissolved in tetrahydrofuran (5 mL) and water (1 mL) followed by addition of lithium hydroxide monohydrate (9 mg,0.21 mmol). The reaction was stirred at room temperature for 1h. The solvent was evaporated after completion of the reaction to give a crude product which was purified by preparation (0.1% formic acid/acetonitrile/water) to give 3- { 2-fluoro-5- [ (isoquinolin-8-ylmethyl) amino ] -4-methoxyphenyl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-062 (10 mg, white solid), yield: 27%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-063)
First step
Preparation of 4-chloro-2-fluoro-6-nitrophenol
Fuming nitric acid (3.97 g,40.92 mmol) was added dropwise to a solution of 4-chloro-2-fluorophenol 146a (5.00 g,34.10 mmol) in acetic acid (30 mL) at 0deg.C. After the completion of the dropwise addition, the reaction solution was stirred at 0℃for 1 hour. The reaction solution was slowly poured into water to precipitate a solid, which was filtered, washed with water and the filter cake was dried to give 4-chloro-2-fluoro-6-nitrophenol 146b (5.89 g, yellow solid), yield: 90%.
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
Preparation of 4-chloro-2-fluoro-6-nitrophenyl triflate
4-chloro-2-fluoro-6-nitrophenol 146b (5.89 g,30.80 mmol) was dissolved in dichloromethane (30 mL), triethylamine (4.67 g,46.20 mmol) was added dropwise, and trifluoromethanesulfonic anhydride (8.69 g,30.80 mmol) was then stirred at 0℃for 2 hours. After the reaction was completed, water was added to quench, and extracted with dichloromethane (3×100 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=20/1) to give 4-chloro-2-fluoro-6-nitrophenyl triflate 146c (9.54 g, white solid), yield: 96%.
1 H NMR(400MHz,CDCl 3 )δ7.99(t,J＝2.2Hz,1H),7.63(dd,J＝8.8,2.5Hz,1H).
Third step
Preparation of 5-chloro-1-fluoro-2-methyl-3-nitrobenzene
To a solution of 4-chloro-2-fluoro-6-nitrophenyl trifluoromethane sulfonate 146c (9.54 g,29.48 mmol) in dioxane (60 mL) was added methylboronic acid (2.12 g,35.49 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (2.16 g,2.96 mmol) and potassium carbonate (6.12 g,44.26 mmol), and stirred at 90℃for 8 hours. After the reaction was completed, water was added to quench, and extracted with ethyl acetate (3×100 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=20/1) to give 5-chloro-1-fluoro-2-methyl-3-nitrobenzene 146d (1.81 g, white solid), yield: 32%.
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).
Fourth step
Preparation of 5-chloro-3-fluoro-2-methylaniline
To a solution of 5-chloro-1-fluoro-2-methyl-3-nitrobenzene 146d (1.81 g,9.55 mmol) in ethanol (10 mL) and water (10 mL) was added ammonium chloride (1.53 g,28.64 mmol) and iron powder (1.60 g,28.64 mmol), and the mixture was stirred at 80℃for 1 hour. After the reaction was completed, the filtrate was filtered while it is hot, 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 5-chloro-3-fluoro-2-methylaniline 146e (1.00 g, colorless liquid), yield: 66%.
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).
Fifth step
Preparation of 5-chloro-7-fluoro-8-methylquinoline
To a solution of 5-chloro-3-fluoro-2-methylaniline 146e (1.00 g,6.30 mmol) and sodium iodide (0.47 g,3.10 mmol) in concentrated sulfuric acid (20 mL) was slowly added glycerol (3.48 g,37.80 mmol) at 140℃and the reaction was stirred at 140℃for 10 hours. After the reaction was completed, the reaction solution was slowly added to methanol for quenching, filtration, concentration of the filtrate, addition of an ammonia water adjustment system to basicity, extraction with ethyl acetate (3×30 mL), drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate, purification by column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) gave 5-chloro-7-fluoro-8-methylquinoline 146f (120 mg, yellow solid), yield: 10%.
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).
Sixth step
Preparation of 8- (bromomethyl) -5-chloro-7-fluoroquinoline
To a solution of 5-chloro-7-fluoro-8-methylquinoline 146f (120 mg,0.61 mmol) in carbon tetrachloride (10 mL) was added 2,2' -azobis (2-methylpropanenitrile) (10 mg,0.06 mmol) and N-bromosuccinimide (120 mg,0.67 mmol), and the mixture was stirred at 80℃for 3 hours. After the reaction was completed, water was added to quench, extracted with methylene chloride (3×20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 146g (104 mg, white solid) of 8- (bromomethyl) -5-chloro-7-fluoroquinoline, yield: 62%.
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 (85 mg,2.47 mmol) in acetonitrile (10 mL) was added potassium carbonate (78 mg,0.57 mmol) and 8- (bromomethyl) -5-chloro-7-fluoroquinoline 146g (104 mg,0.38 mmol), and the reaction mixture was stirred at 70℃for 2 hours. 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 column chromatography (mobile phase: petroleum ether/ethyl acetate=4/1) to give 5-chloro-7-fluoro-8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoline 146h (135 mg, white solid), yield: 93%.
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 for 146h (135 mg,0.35 mmol) in ethanol (5 mL) and water (5 mL) was added iron powder (59 mg,1.06 mmol) and ammonium chloride (57 mg,1.06 mmol) and the reaction mixture was stirred at 80℃for 1 h. After the reaction was completed, the filtrate was filtered while it was still hot, extracted with ethyl acetate (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, 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%.
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).
Ninth step
Preparation of dimethyl 4- [ ({ 5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
To a solution of 5- [ (5-chloro-7-fluoroquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 146i (90 mg,0.26 mmol) in tetrahydrofuran (5 mL) was added triethylamine (39 mg,0.38 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (103 mg,0.31 mmol), stirred at 25 ℃ for 4 hours, after completion of the reaction, water quench was added, extraction was performed 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 column chromatography (mobile phase: dichloromethane/methanol=10/1) to give dimethyl 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).
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-dicarboxylic acid 146j (106 mg,0.18 mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (13 mg,0.54 mmol), and the mixture was reacted at 25℃for 2 hours. After completion of the reaction, 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 via a 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%.
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)
First step
Preparation of 5-fluoro-4-methyl-1, 3-benzothiazol-2-amine
3-fluoro-2-methylaniline 147a (10 g,0.08 mol) and potassium thiocyanate (31.06 g,0.32 mol) were dissolved in acetic acid (100 mL) and stirred at room temperature for 30min before Br was added 2 (12.64 g,0.0791 mol) was diluted with acetic acid (30 mL) and added dropwise to the above solution under an ice bath. The reaction solution was reacted at room temperature for 5 hours. After the reaction was completed, it was quenched with water, ph=8 to 9 was adjusted with sodium hydroxide solution, and then extracted with ethyl acetate (3×30 mL). The organic phases are combined and then treated with anhydrous sodium sulfateDrying, filtration and purification of the concentrated residue by column chromatography (petroleum ether/ethyl acetate: 99/1) gave 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
Preparation of 2-bromo-5-fluoro-4-methyl-1, 3-benzothiazole
5-fluoro-4-methyl-1, 3-benzothiazol-2-amine 147b (8 g,0.04 mol) was dissolved in acetonitrile (100 mL). Cuprous bromide (2.5 g,0.04 mol) and tert-butyl nitrite (13.58 g,0.1317 mol) were added sequentially. The reaction solution was reacted at 50℃for 3 hours. After the completion of the reaction, the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 99/1) to give 147c (9 g, yellow solid) of 2-bromo-5-fluoro-4-methyl-1, 3-benzothiazole in 8% yield.
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).
Third step
Preparation of 2-bromo-4- (bromomethyl) -5-fluoro-1, 3-benzothiazole
2-bromo-5-fluoro-4-methyl-1, 3-benzothiazole 147c (300 mg,1.22 mmol) was dissolved in carbon tetrachloride (10 mL) and benzoyl peroxide (148 mg,0.61 mmol) and N-bromosuccinimide (304 mg,1.71 mmol) were added at room temperature. The reaction mixture was reacted at 85℃for 3 hours. After the completion of the reaction, the reaction mixture was concentrated and subjected to column chromatography (mobile phase: petroleum ether/ethyl acetate=99/1) to give 147d (100 mg, white solid) of 2-bromo-4- (bromomethyl) -5-fluoro-1, 3-benzothiazole in 17.67% yield.
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).
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 (600 mg,1.84 mmol) was dissolved in acetonitrile (30 mL), 4-fluoro-2-methoxy-5-nitrophenol (345 mg,1.84 mmol), potassium carbonate (510 mg,3.69 mmol) and potassium iodide (459 mg,2.77 mmol) were added sequentially, 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)。
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 (110 mg,0.25 mmol) and methylphenylboronic acid (7 mg,0.12 mmol) were dissolved in toluene/ethanol/water (2 mL, 7:2:1). Potassium carbonate (105 mg,0.76 mmol) and tetrakis (triphenylphosphine) palladium (88 mg,0.07 mmol) were then added 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, 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 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).
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 (160 mg,0.45 mmol) was dissolved in acetonitrile (2 mL). Then, a saturated ammonium chloride solution (2 mL) and iron powder (508 mg,9.08 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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-dicarboxylic acid
147g (135 mg,0.42 mmol) of 2-fluoro-5- [ (5-fluoro-1, 3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (280 mg,0.83 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (423.78 mg,4.188 mmol) was then added and the reaction was reacted at 25℃for 16 hours. The reaction solution was not subjected to any post-treatment to obtain dimethyl 4- [ ({ 2-fluoro-5- [ (5-fluoro-1, 3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 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
Dimethyl 4- [ ({ 2-fluoro-5- [ (5-fluoro-1, 3-benzothiazol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 147h (5 mL stock solution) with lithium hydroxide (50 mg,2.12 mmol) was dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) to give 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, pale yellow solid), yield: 54%.
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)
First step
Preparation of 1-benzoyl-3- (3-chloro-2-methylphenyl) thiourea
3-chloro-2-methylaniline 148a (10 g,70.6 mmol) was dissolved in tetrahydrofuran (90 mL). Benzoyl isothiocyanate (23.04 g,141.2 mmol) was then added at 25 ℃. The reaction solution was reacted at 25℃for 16 hours. Then spin-dry the solvent followed by ethanol: water = 4:1 (50 mL) was filtered 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
Preparation of (3-chloro-2-methylphenyl) thiourea
1-benzoyl-3- (3-chloro-2-methylphenyl) thiourea 148b (20 g,65.6 mmol) was dissolved in methanol: water = 3:1 (60 mL). Potassium carbonate (27.20 g,196.8 mmol) was then added at 25 ℃. The reaction solution was reacted at 70℃for 2 hours. Then spin-drying the solvent, adding water, and filtering to obtain residue to obtain (3-chloro-2-methylphenyl) thiourea 148c (11 g, white solid) with a yield of 75%.
MS m/z(ESI)：201[M+1] + 。
Third step
Preparation of 5-chloro-4-methyl-1, 3-benzothiazol-2-amine
(3-chloro-2-methylphenyl) thiourea 148c (12 g,59.8 mmol) was dissolved in chloroform (50 mL), and then liquid bromine (11.47 g,71.7 mmol) was added dropwise at 25 ℃. The reaction solution was reacted at 60℃for 2 hours. Then spin-drying the solvent, adding water, and filtering to obtain filter residue to obtain 5-chloro-4-methyl-1, 3-benzothiazol-2-amine 148d (11 g, white solid) with the yield of 83%.
MS m/z(ESI)：199[M+1] + 。
Fourth step
Preparation of 5-chloro-4-methyl-1, 3-benzothiazole
5-chloro-4-methyl-1, 3-benzothiazol-2-amine 148d (1.3 g, 0.006mol) was dissolved in 1, 4-dioxane (20 mL). Tert-butyl nitrite (2.01 g,0.01 mol) was then 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] + 。
Fifth step
Preparation of 4- (bromomethyl) -5-chloro-1, 3-benzothiazole
5-chloro-4-methyl-1, 3-benzothiazole 148e (1.2 g,1.80 mmol) was dissolved in carbon tetrachloride (15 mL). Dibenzoyl peroxide (0.79 g,3.2 mmol) was then added at 25℃with N-bromosuccinimide [1.39g,7.8mmol ], and the reaction was allowed to react 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] + 。
Sixth step
Preparation of 5-chloro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole
4- (bromomethyl) -5-chloro-1, 3-benzothiazole 148e (900 mg,3.43 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (769 mg,4.11 mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (0.947 g,6.85 mmol) and potassium iodide (0.853 g,5.14 mmol) were added, and the reaction mixture was reacted at 65℃for 2 hours. The solvent was then dried by spinning and extracted with dichloromethane (2X 10 mL). The combined organic phases were 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 (900 mg,2.44 mmol) was dissolved in acetonitrile (10 mL). Then, a saturated ammonium chloride solution (10 mL) and iron powder (408 mg,7.32 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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 dimethyl 4- [ ({ 5- [ (5-chloro-1, 3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
148g (200 mg,0.59 mmol) of 5- [ (5-chloro-1, 3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (292 mg,0.88 mmol) were dissolved in tetrahydrofuran (10 mL.) then triethylamine (178 mg,1.77 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：580.0[M+1] + 。
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
Dimethyl 4- [ ({ 5- [ (5-chloro-1, 3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 148h (10 mL stock solution) with lithium hydroxide (21 mg,0.078 mmol) was dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=5:95) to give 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), yield: 47%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-066)
First step
Preparation of 2-amino-5-chloro-3-fluorobenzoic acid
To a solution of 2-amino-3-fluorobenzoic acid 149a (10.00 g,64.50 mmol) in N, N-dimethylformamide (60 mL) was added N-chlorosuccinimide (10.34 g,77.40 mmol), and the reaction was stirred at 25℃for 16 hours. After the reaction was completed, water was added to quench and filter. The filter cake was dried to give 2-amino-5-chloro-3-fluorobenzoic acid 149b (11.0 g, yellow solid), yield: 90%.
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
Preparation of methyl 2-amino-5-chloro-3-fluorobenzoate
To a solution of 2-amino-5-chloro-3-fluorobenzoic acid 149b (11.00 g,58.00 mmol) in N, N-dimethylformamide (100 mL) were added methyl iodide (8.83 g,69.60 mmol) and potassium carbonate (12.02 g,87.00 mmol), and the reaction was stirred at 25℃for 16 hours. After the reaction was completed, water was added to quench and filter. The filter cake was dried to give methyl 2-amino-5-chloro-3-fluorobenzoate 149c (10.00 g, pink solid), yield: 85%.
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).
Third step
Preparation of methyl 2-bromo-5-chloro-3-fluorobenzoate
To a solution of methyl 2-amino-5-chloro-3-fluorobenzoate 149c (10.00 g,49.10 mmol) in acetonitrile (60 mL) was added dropwise an aqueous 40% hydrogen bromide solution (60 mL) and an aqueous sodium nitrite solution (4.07 g,58.90 mmol) under nitrogen protection at 0℃and, after 10 minutes of reaction, cuprous bromide (8.45 g,58.90 mmol) was added in portions. After the completion of the addition, the reaction was continued at 0℃for 1 hour, and after the completion of the reaction, water was added for dilution and extraction with ethyl acetate (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: petroleum ether/ethyl acetate=20/1) to give methyl 2-bromo-5-chloro-3-fluorobenzoate 149d (9.60 g, white solid), yield: 73%.
1 H NMR(400MHz,CDCl 3 )δ760-7.59(m,1H),7.29–7.27(m,1H),3.96-3.95(m,3H).
Fourth step
Preparation of (2-bromo-5-chloro-3-fluorophenyl) methanol
To a solution of methyl 2-bromo-5-chloro-3-fluorobenzoate 149d (9.76 g,36.50 mmol) in tetrahydrofuran (60 mL) was added dropwise a 1N solution of diisobutylaluminum hydride in N-hexane (40 mL) under nitrogen protection at 0℃and the reaction was stirred at 0℃for 2 hours. After the reaction was completed, water and saturated sodium bicarbonate solution were added to quench and filter, 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), yield: 87%.
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).
Fifth step
Preparation of 2-bromo-5-chloro-3-fluorobenzaldehyde
To a solution of (2-bromo-5-chloro-3-fluorophenyl) methanol 149e (7.60 g,31.70 mmol) in methylene chloride (60 mL) was added Dess-Martin reagent (17.48 g,41.20 mmol) and the reaction was stirred at 25℃for 0.5 h. After the reaction was completed, a saturated aqueous sodium hydrogensulfite solution and a saturated sodium hydrogencarbonate solution were added thereto, and the mixture was quenched with methylene chloride (3X 100 mL) and extracted with the filtrate. 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), yield: 86%.
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).
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 (6.50 g,27.40 mmol) and 2, 2-dimethoxyethylamine (3.46 g,32.80 mmol) in ethanol (60 mL) was stirred at 80℃for 8 hours, then sodium cyanoborohydride (2.07 g,32.80 mmol) and acetic acid (2 mL) were added and reacted at 25℃for 1 hour. After completion of the reaction, quench by adding saturated aqueous ammonium chloride, extract with ethyl acetate (3×100 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify by column chromatography (mobile phase: petroleum ether/ethyl acetate=10/1) to give [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] (2, 2-dimethoxyethyl) amine 149g (8.00 g, yellow oil), yield: 89%.
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 [ (2-bromo-5-chloro-3-fluorophenyl) methyl ] (2, 2-dimethoxyethyl) amine 149g (8.60 g,26.30 mmol) and triethylamine (3.99 g,39.40 mmol) in methylene chloride (80 mL) was added dropwise p-toluenesulfonyl chloride (5.52 g,28.90 mmol) at 0℃and reacted at 0℃for 3 hours. After completion of the reaction, quench with water (100 mL), extract with dichloromethane (3×100 mL), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate and purify 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%.
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 (15.20 g,114.00 mmol) 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 149h (11.00 g,22.80 mmol) in methylene chloride (60 mL) at 0℃and, after completion of the addition, the reaction was carried out at 0℃for 8 hours. After the reaction was completed, the reaction solution was slowly dropped to ice water for quenching, and extracted with methylene chloride (3×100 mL). The organic phase was washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and 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), yield: 48%.
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).
Ninth step
Preparation of 5-chloro-7-fluoroisoquinoline-8-carboxylic acid methyl ester
To a solution of 8-bromo-5-chloro-7-fluoroisoquinoline 149i (1.00 g,3.80 mmol) and triethylamine (580 mg,5.70 mmol) in methanol (20 mL) was added [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) (280 mg,0.30 mmol), and the mixture was reacted at 80℃under 1atm carbon monoxide for 16 hours. After completion of the reaction, water (20 mL) was added thereto, followed by extraction 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-chloro-7-fluoroisoquinoline-8-carboxylate 149j (653 mg, white solid), yield: 71%.
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).
Tenth step
Preparation of (5-chloro-7-fluoroisoquinolin-8-yl) methanol
To a solution of 5-chloro-7-fluoroisoquinoline-8-carboxylic acid methyl ester 149j (353 mg,1.47 mmol) in tetrahydrofuran (10 mL) was added dropwise a 1N solution of lithium aluminum hydride in tetrahydrofuran (2.2 mL) at 0℃under nitrogen protection, and the mixture was reacted at 0℃for 2 hours. After completion of the reaction, water was added to quench (5 mL), and 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 column chromatography (mobile phase: dichloromethane/methanol=10/1) to give (5-chloro-7-fluoroisoquinolin-8-yl) methanol 149k (300 mg, yellow solid), yield: 96%.
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).
Eleventh step
Preparation of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline
To a solution of (5-chloro-7-fluoroisoquinolin-8-yl) methanol 149k (358 mg,1.69 mmol) in methylene chloride (10 mL) was added dropwise phosphine tribromide (687 mg,2.54 mmol) at 0℃and the reaction was carried out at 0℃for 2 hours. After the reaction was completed, a saturated aqueous sodium hydrogencarbonate solution was added to adjust ph=6 to 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 (380 mg, yellow oil) of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline, yield: 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 (311 mg,1.66 mmol) and potassium carbonate (287 mg,2.08 mmol) in acetonitrile (10 mL) was added 149l (380 mg,1.38 mmol) of 8- (bromomethyl) -5-chloro-7-fluoroisoquinoline, and the mixture was reacted at 70℃for 3 hours. After completion of the reaction, water (10 mL) was added thereto, followed by extraction 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), yield: 35%.
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 (157 mg,0.41 mmol) in ethanol (4 mL) and water (4 mL) was added iron powder (69 mg,1.24 mmol) and ammonium chloride (66 mg,1.24 mmol), and reacted at 80℃for 1 hour. After completion of the reaction, extracted with ethyl acetate (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 149n (104 mg, yellow solid), yield: 72%.
MS m/z(ESI)：351.1[M+1] + 。
Fourteenth step
Preparation of dimethyl 4- [ ({ 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
To a solution of 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyaniline 149n (100 mg,0.29 mmol) and triethylamine (43 mg,0.43 mmol) in tetrahydrofuran (5 mL) was added dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (115 mg,0.34 mmol), and after reaction at 25℃for 5 hours, water (5 mL) was added to quench, extract 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 dimethyl 4- [ ({ 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate 149o (110 mg, white solid) in a yield of 65%.
MS m/z(ESI)：592.1[M+1] + 。
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 dimethyl 4- [ ({ 5- [ (5-chloro-7-fluoroisoquinolin-8-yl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 149o (100 mg,0.17 mmol) in tetrahydrofuran (2 mL), methanol (2 mL) and water (2 mL) was added lithium hydroxide (12 mg,0.51 mmol) and reacted 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 via a 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%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-067)
First step
Preparation of N- ((5-chloro-2-methylphenyl) aminomethyl) benzamide
5-chloro-2-methylaniline 150a (2.0 g,14.1 mmol) was dissolved in tetrahydrofuran (100 mL) followed by addition of benzoyl isothiocyanate (3.45 g,21.1 mmol). The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was concentrated to give N- ((5-chloro-2-methylphenyl) aminomethylsulfonyl) benzamide 150b (4 g, yellow solid) in 89% yield.
MS m/z(ESI)：305.0[M+1] + 。
Second step
Preparation of 1- (5-chloro-2-methylphenyl) thiourea
N- ((5-chloro-2-methylphenyl) aminomethyl) benzamide 150b (4 g,13.1 mmol) was dissolved in methanol (60 mL), then an aqueous solution (20 mL) of sodium carbonate (2.78 g,26.2 mmol) was added, and the reaction solution was stirred at 70℃for 2 hours. After the completion of the reaction, the crude product was concentrated, followed by extraction with methylene chloride (3X 30 mL), and the organic phase was dried over saturated brine, anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure by filtration to give compound 1- (5-chloro-2-methylphenyl) thiourea 150c (2.57 g, yellow oil) in 98% yield.
MS m/z(ESI)：201.1[M+1] + 。
Third step
Preparation of 7-chloro-4-methylbenzo [ d ] thiazol-2-amine
1- (5-chloro-2-methylphenyl) thiourea 150c (2.57 g,12.8 mmol) was dissolved in chloroform (100 mL), and bromine (2.04 g,12.8 mmol) was added thereto and the reaction was stirred at 70℃for 2 hours. After the completion of the reaction, an aqueous sodium thiosulfate solution (50 mL) was added, the organic phase was separated after washing with water, the aqueous phase was extracted with methylene chloride (3×30 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] + 。
Fourth step
Preparation of 7-chloro-4-methylbenzo [ d ] thiazole
7-Chlorobenzene-4-methyl-o [ d ] thiazol-2-amine 150d (1.8 g,9.06 mmol) was dissolved in tetrahydrofuran (100 mL), tert-butyl nitrite (2.82 g,27.3 mol) was added, and the reaction mixture was stirred at 70℃for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by a 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] + 。
Fifth step
Preparation of 4- (bromomethyl) -7-chlorobenzo [ d ] thiazole
7-chloro-4-methylbenzo [ d ] thiazole 150e (400 mg,2.18 mmol) was dissolved in carbon tetrachloride (50 mL), and then N-bromosuccinimide (426.4 mg,2.39 mmol) and azobisisobutyronitrile (35.8 mg,0.218 mmol) were added, respectively, and the reaction solution was stirred at 80℃for 3 hours. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by a silica gel column (petroleum ether/ethyl acetate=20/1) to give 4- (bromomethyl) -7-chlorobenzo [ d ] thiazole 150f (430 mg, white solid) in 71% yield.
MS m/z(ESI)：263.9[M+1] + 。
Sixth step
Preparation of 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole
4- (bromomethyl) -7-chlorobenzo [ d ] thiazole 150f (200 mg,0.76 mmol)), 4-fluoro-2-methoxy-5-nitroaniline (171 mg,0.91 mmol), potassium carbonate (211 mg,1.52 mmol) and potassium iodide (63 mg,0.38 mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50℃for 2 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=23%) afforded 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole 150g (210 mg, white solid), yield: 71%.
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 (85 mg,0.23 mmol) of 7-chloro-4- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -benzo [ d ] thiazole, iron powder (38 mg,0.69 mmol) and ammonium chloride (37 mg,0.69 mmol) were dissolved in ethanol: water = 1:1 (12 mL), the reaction 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 for 150h (80 mg, brown solid), yield: 97%. The crude product was used directly in the next reaction.
MS m/z(ESI)：339.1[M+1] + 。
Eighth step
Preparation of dimethyl 4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
5- ((7-Chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyaniline (80 mg,0.24 mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (95 mg,0.28 mmol) and triethylamine (47 mg,0.47 mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was distilled off to give crude product, dimethyl 4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 150i (70 mg, yellow solid), yield: 48%. The crude product was used directly in the next reaction.
MS m/z(ESI)：580.0[M+1] + 。
Ninth step
Preparation of 3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 150i (60 mg,0.10 mmol) was dissolved in tetrahydrofuran/water=1: 1 (10 mL) and then lithium hydroxide (8 mg,0.31 mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture 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 preparation of (FA) to give 3- (5- ((7-chlorobenzo [ d ] thiazol-4-yl) methoxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-067 (43.03 mg, yellow solid), yield: 76%.
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)
First step
Preparation of 1-benzoyl-3- (5-fluoro-2-methylphenyl) thiourea
5-fluoro-2-methylaniline 151a (2 g,16 mmol) was dissolved in tetrahydrofuran (20 mL), and benzoyl isothiocyanate (5.22 g,32 mmol) was added dropwise to the solution. The reaction was stirred at 25℃for 16 hours under air protection. After the reaction was completed, 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- (5-fluoro-2-methylphenyl) thiourea 151b (2.8 g, white solid), yield: 59%.
MS m/z(ESI)：289.0[M+1] + 。
Second step
Preparation of (5-fluoro-2-methylphenyl) thiourea
1-benzoyl-3- (5-fluoro-2-methylphenyl) thiourea 151b (2.8 g,9.7 mmol) and potassium carbonate (2.68 g,19.4 mmol) were dissolved in methanol: water=4:1 (30 mL), and the reaction solution was stirred at 70℃for 2 hours. After the reaction was completed, 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). The filter cake was (5-fluoro-2-methylphenyl) thiourea 151c (1.8 g, white solid), yield: 96%.
MS m/z(ESI)：185.1[M+1] + 。
Third step
Preparation of 7-fluoro-4-methyl-1, 3-benzothiazol-2-amine
(5-fluoro-2-methylphenyl) thiourea 151c (1.8 g,0.9 mmol) was dissolved in chloroform (30 mL), and bromine (2.35 g,14.7 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 (30 mL), to which was added an aqueous solution of sodium thiosulfate (20 mL). 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%.
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).
Fourth step
Preparation of 7-fluoro-4-methyl-1, 3-benzothiazole
7-fluoro-4-methyl-1, 3-benzothiazol-2-amine 151d (890 mg,4.88 mmol) was dissolved in 1, 4-dioxane (30 mL), followed by dropwise addition of tert-butyl nitrite (1000 mg,9.77 mmol) to the solution at room temperature. The mixture was stirred at 90℃for 1 hour. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=5%) gave 7-fluoro-4-methyl-1, 3-benzothiazole 151e (440 mg, yellow liquid), yield: 53%.
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).
Fifth step
Preparation of 4- (bromomethyl) -7-fluoro-1, 3-benzothiazole
7-fluoro-4-methyl-1, 3-benzothiazole 151e (440 mg,2.63 mmol), N-bromosuccinimide (560 mg,3.16 mmol) and azobisisobutyronitrile (216 mg,2.16 mmol) were dissolved in carbon tetrachloride (10 mL). The reaction solution was stirred at 85℃for 16 hours under a nitrogen atmosphere. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=5%) afforded 4- (bromomethyl) -7-fluoro-1, 3-benzothiazole 151f (270 mg, yellow solid), yield: 41%.
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).
Sixth step
Preparation of 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole
4- (bromomethyl) -7-fluoro-1, 3-benzothiazole 151f (270 mg,1.09 mmol), 4-fluoro-2-methoxy-5-nitroaniline (246 mg,1.31 mmol), potassium carbonate (303 mg,2.19 mmol) and potassium iodide (91 mg,0.54 mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50℃for 2 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=23%) afforded 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole 151g (270 mg, white solid), yield: 68%.
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
151g (150 mg,0.43 mmol) of 7-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole, iron powder (71 mg,1.27 mmol) and ammonium chloride (68 mg,1.27 mmol) were dissolved 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), ethyl acetate (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 2-fluoro-5- [ (7-fluoro-1, 3-benzothiazol-4-yl) methoxy ] -4-methoxyaniline 151h (130 mg, brown solid), yield: 71%. The crude product was used directly in the next reaction.
MS m/z(ESI)：323.1[M+1] + 。
Eighth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((7-fluorobenzo [ 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 (130 mg,0.40 mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (162 mg,0.48 mmol) and triethylamine (81 mg,0.81 mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was completed, crude 4- (3- (2-fluoro-5- ((7-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid dimethyl ester 151i was used directly in the next step without treatment.
MS m/z(ESI)：564.0[M+1] + 。
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
Dimethyl 4- (3- (2-fluoro-5- ((7-fluorobenzo [ d ] thiazol-4-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 151i was dissolved in tetrahydrofuran/methanol/water=1: 1:1 (6 mL) and then lithium hydroxide (7 mg,0.27 mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture 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 preparation of (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%.
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-benzoxazol-5-yl } -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-069)
First step
Preparation of 2-amino-6-methoxy-4-nitrophenol
2-methoxy-4, 6-dinitrophenol 155a (1.00 g,4.67 mmol) was dissolved in ethanol (20 mL) and then tin dichloride (2.69 g,14.01 mmol) was added. The reaction was stirred at 90℃for 2 hours. After LCMS detection reaction was completed, the reaction was cooled to room temperature. After removal of the solvent by distillation under reduced pressure, purification by silica gel column (petroleum ether/ethyl acetate=1/1) gave the product 2-amino-6-methoxy-4-nitrophenol 155b (300 mg, yellow solid), yield: 31%.
MS m/z(ESI)：185.1[M+1] + 。
Second step
Preparation of 7-methoxy-5-nitro-1, 3-benzoxazole
2-amino-6-methoxy-4-nitrophenol 155b (150 mg,0.81 mmol) was dissolved in trimethyl orthoformate (10 mL). The reaction was refluxed at 90 ℃ for 2 hours. After LCMS monitored the reaction was complete, the reaction was cooled to room temperature and the solvent was distilled off under reduced pressure. Purification over a silica gel column (petroleum ether/ethyl acetate=1/1) gave the product 7-methoxy-5-nitro-1, 3-benzoxazole 155c (100 mg, yellow solid), yield: 60%.
MS m/z(ESI)：195.1[M+1] + 。
Third step
Preparation of 5-nitro-1, 3-benzoxazol-7-ols
7-methoxy-5-nitro-1, 3-benzoxazole 155c (120 mg,0.62 mmol) was dissolved in DMF (5 mL) followed by the addition of lithium chloride (52 mg,1.24 mmol). The reaction was stirred in a microwave reactor at 160℃for 2h. After completion of the reaction, 5-nitro-1, 3-benzoxazol-7-ol 155d (80 mg, yellow solid) was obtained via reverse column (water/acetonitrile=3/7), yield: 68%.
MS m/z(ESI)：181.1[M+1] + 。
Fourth step
Preparation of 7- [ ((2, 3-difluoro-6-methoxyphenyl) methoxy ] -5-nitro-1, 3-benzoxazole
5-nitro-1, 3-benzooxazol-7-ol 155d (80 mg,0.44 mmol) and 2- (chloromethyl) -3, 4-difluoro-1-methoxybenzene (94 mg,0.48 mmol) were dissolved in acetonitrile (20 mL), followed by the addition of potassium carbonate (122 mg,0.88 mmol) and potassium iodide (36 mg,0.22 mmol). The reaction solution was stirred at 50℃for 4h. 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 a 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), yield: 26%.
MS m/z(ESI)：337.1[M+1] + 。
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 (45 mg,0.13 mmol) was dissolved in acetonitrile (10 mL) and saturated ammonium chloride (2 mL), followed by addition of iron powder (74 mg,1.34 mmol). The reaction was stirred at 25℃for 0.5h, the iron powder was filtered after completion of the reaction, 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-benzoxazol-5-amine 155f (40 mg, yellow solid) in 63% yield.
MS m/z(ESI)：307.1[M+1] + 。
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-benzoxazol-5-amine 155f (40 mg,0.13 mmol) was dissolved in tetrahydrofuran (5 mL), followed by addition of dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (52 mg,0.15 mmol) and triethylamine (39 mg,0.39 mmol). After completion of the LCMS monitoring reaction, the reaction solution was stirred at 25℃for 16 hours without any post-treatment to give 155g of dimethyl 4- [ ({ - [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -1, 3-benzoxazol-5-yl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate as a stock solution.
MS m/z(ESI)：548.1[M+1] + 。
Seventh step
Preparation of 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
Dimethyl 4- [ ({ - [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -1, 3-benzoxazol-5-yl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 155g (5 mL stock solution) with lithium hydroxide (5 mg,0.11 mmol) was dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and subjected to preliminary purification by reverse column (water: acetonitrile=20:80) to give 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) in 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-070)
First step
Preparation of 2-methylquinoline-8-carboxylic acid methyl ester
8-bromo-2-methylquinoline 156a (1400 mg,6.3 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1383 mg,1.9 mmol) and triethylamine (1913 mg,18.9 mmol) were dissolved in methanol (40 mL), and then the reaction solution was stirred under carbon monoxide (1.5 MPa) atmosphere at 70℃for 8 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=20%) gave methyl 2-methylquinoline-8-carboxylate 156b (1300 mg, grey solid), yield: 92%.
MS m/z(ESI)：202.1[M+1] + 。
Second step
Preparation of (2-methylquinolin-8-yl) methanol
Methyl 2-methylquinoline-8-carboxylate 156b (600 mg,2.7 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), followed by dropwise hydrogenation of a lithium aluminum solution (1.0M in tetrahydrofuran) (3 mL) at 0deg.C. The mixture was stirred at room temperature for 1 hour. 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-methylquinolin-8-yl) methanol 156c (300 mg, gray solid), yield: 56%.
MS m/z(ESI)：196.1[M+1] + 。
Third step
Preparation of 8- (bromomethyl) -2-methylquinoline
(2-Methylquinolin-8-yl) methanol 156c (300 mg,1.53 mmol) was dissolved in methylene chloride (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), yield: 46%.
MS m/z(ESI)：236.0[M+1] + . Fourth step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methylquinoline
8- (bromomethyl) -2-methylquinoline 156d (100 mg,0.39 mmol)), 4-fluoro-2-methoxy-5-nitroaniline (250 mg,1.40 mmol), potassium carbonate (1011 mg,5.4 mmol) and potassium iodide (52 mg,0.21 mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50℃for 2 hours. After the completion of the reaction, 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=50%) afforded 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methylquinoline 156e (55 mg, yellow solid), yield: 40%.
MS m/z(ESI)：343.1[M+1] + 。
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 (55 mg,0.15 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of iron powder (650 mg,9.4 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 (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%. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.1[M+1] + 。
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 (50 mg,0.13 mmol) was dissolved in tetrahydrofuran (6 mL), followed by the addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (686 mg,2.0 mmol) and triethylamine (162 mg,1.52 mmol) 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%. 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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
156g (65 mg,0.11 mmol) 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) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the completion of the reaction, 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. The crude product was purified by preparation of (FA) to give 3- (2-fluoro-4-methoxy-5- ((2-methylquinolin-8-yl) methoxy) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-070 (12 mg, yellow solid), yield: 20%.
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)
First step
Preparation of 8-bromo-3-methylquinoline
2-Bromoaniline 157a (10 g,5.8 mmol) and 2-methylprop-2-enal (6.12 g,8.8 mmol) were dissolved in 6N hydrochloric acid (10 mL). The reaction solution was stirred at 110℃for 16 hours under nitrogen protection. After the reaction was completed, the reaction mixture was quenched with 5N sodium hydroxide solution to pH >9, extracted with methylene chloride (3X 20 mL), and the organic phase was dried over anhydrous sodium sulfate. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=7%) gave 8-bromo-3-methylquinoline 157b (2.5 g, yellow liquid), yield: 19%.
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
Preparation of 3-methylquinoline-8-carboxylic acid methyl ester
8-bromo-3-methylquinoline 157b (1.5 g,6.8 mmol) and 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride (2.49 g,3.4 mmol) were dissolved in methanol (10 mL), triethylamine (1.38 g,13.6 mmol) was added at room temperature, and then the reaction solution was stirred under carbon monoxide (1.5 MPa) atmosphere at 100deg.C for 8 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=30%) gave methyl 3-methylquinoline-8-carboxylate 157c (270 mg, yellow solid), yield: 20%.
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).
Third step
Preparation of (3-methylquinolin-8-yl) methanol
3-methylquinoline-8-carboxylic acid methyl ester 157c (100 mg,0.50 mmol) was dissolved in tetrahydrofuran (10 mL), and lithium diisobutylhydride (0.5 mL,0.50 mmol) was added dropwise at-78 ℃. The reaction solution was stirred at-78℃for 1 hour. After the completion of the reaction, the reaction solution was quenched with saturated potassium sodium 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 a crude product (3-methylquinolin-8-yl) methanol 157d (90 mg, yellow solid), yield: 94%. The crude product was used directly in the next reaction.
MS m/z(ESI)：174.3[M+1] + 。
Fourth step
Preparation of 8- (bromomethyl) -3-methylquinoline
(3-Methylquinolin-8-yl) methanol 157d (70 mg,0.40 mmol) was dissolved in methylene chloride (5 mL), and phosphorus tribromide (33 mg,0.12 mmol) was added dropwise to the solution at 0 ℃. The mixture was stirred at 0℃for 1 hour. After the completion of the reaction, the reaction mixture was quenched with water (10 mL), extracted with methylene chloride (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration gave crude 8- (bromomethyl) -3-methylquinoline 157e (73 mg, yellow solid), yield: 72%. The crude product was used directly in the next reaction.
MS m/z(ESI)：238.1(M+H)。
Fifth step
Preparation of 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline
8- (bromomethyl) -3-methylquinoline 157e (73 mg,0.31 mmol), 4-fluoro-2-methoxy-5-nitroaniline (69 mg,0.37 mmol), potassium carbonate (85 mg,0.62 mmol) and potassium iodide (25 mg,0.15 mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50℃for 2 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=23%) afforded 8- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline 157f (60 mg, white solid), yield: 56%.
MS m/z(ESI)：343.1[M+1] + 。
Sixth step
Preparation of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline
78- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -3-methylquinoline 157f (50 mg,0.15 mmol), iron powder (24 mg,0.44 mmol) and ammonium chloride (23 mg,0.44 mmol) were dissolved in ethanol: water = 1:1 (5 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 (10 mL), ethyl acetate (3×10 mL) was extracted, and the combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 157g of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline (60 mg, brown solid), yield: 86%. The crude product was used directly in the next reaction.
MS m/z(ESI)：313.3[M+1] + 。
Seventh step
Preparation of dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate
157g (65 mg,0.21 mmol) of 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] aniline is dissolved in tetrahydrofuran (5 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (83 mg,0.24 mmol) and triethylamine (42 mg,0.42 mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was completed, it was filtered and concentrated to give dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylate 157h (30 mg, yellow solid), yield: 25%. The crude product was directly used in the next reaction
MS m/z(ESI)：554.0[M+1] + 。
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
Dimethyl 4- [ ({ 2-fluoro-4-methoxy-5- [ (3-methylquinolin-8-yl) methoxy ] phenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 157h (30 mg,0.05 mmol) was dissolved in tetrahydrofuran/water=1: 1 (6 mL) and then lithium hydroxide (4 mg,0.16 mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture 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 preparation of (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%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (pd-072)
First step
Preparation of 2-methoxy-8-methylquinoline
Compound 158a (500 mg,2.81 mmol) and CH 3 ONa (1.52 g,28.1 mmol) in CH 3 OH (10 mL), at 65℃for 48h. The reaction solution 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%.
MS m/z(ESI)：174.0[M+1] + .
Second step
Preparation of 8- (bromomethyl) -2-methoxyquinoline
Compound 158b (100 mg,0.58 mmol), BPO (7 mg,0.03 mmol) and NBS (113 mg,0.64 mmol) were dissolved in CCl 4 (10 mL) was reacted 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%.
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).
Third step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2-methoxyquinoline
Compound 158c (100 mg,0.40 mmol), 4-fluoro-2-methoxy-5-naphthol (82 mg,0.44 mmol) and K 2 CO 3 (165 mg,1.2 mmol) was dissolved in ACN (10 mL) and reacted at 80℃for 2h. The reaction was filtered and 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%.
MS m/z(ESI)：359.1[M+1] + .
Fourth step
Preparation of 2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) aniline
Compound 158d (90 mg,0.25 mmol) was dissolved in MeOH: NH 4 To a solution of cl=5:1 (12 mL), fe powder (140 mg,2.5 mmol) was added and reacted at room temperature for 2h. The reaction solution was filtered, the filtrate was concentrated, and the obtained residue was dissolved in water, extracted with ethyl acetate, washed with saturated NaCl, 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), yield: 96%.
MS m/z(ESI)：329.2[M+1] + .
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 (70 mg,0.21 mmol) and TEA (65 mg,0.63 mmol) were dissolved in dry THF (5 mL) and then 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (107 mg,0.31 mmol) was added and reacted at room temperature for 16h. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (EtOAc: pe=30%) to give dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 158d (60 mg, yellow solid), yield: 45%.
MS m/z(ESI)：570.1[M+1] + .
Sixth step
Preparation of 3- (2-fluoro-4-methoxy-5- ((2-methoxyquinolin-8-yl) methoxy) phenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 158d (60 mg,0.11 mmol) was dissolved in THF/MeOH/H 2 O=3:1:1 (10 mL) and then LiOH (8 mg,0.33 mmol) was added and reacted at room temperature for 1h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (ACN: H) 2 O (0.1% FA) =72%) to obtain 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: 48%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-073)
First step
Preparation of quinoline-8-carboxylic acid methyl ester
Compound 159a (1 g,5.8 mmol) was dissolved in anhydrous DCM (25 mL), oxalyl chloride (1.1 g,8.7 mmol) and a catalytic amount of DMF were added under ice-bath and reacted for 1h at 0deg.C followed by MeOH (10 mL). The reaction was concentrated under reduced pressure and the residue was dissolved in EtOAc (20 mL) and taken up in saturated NaHCO respectively 3 And saturated NaCl water washing, and drying with anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography (EtOAc: pe=50%) to give methyl quinoline-8-carboxylate 159b (0.9 g, orange-yellow oil), yield: 79%.
MS m/z(ESI)：188.0[M+1] + .
Second step
Preparation of 3-hydroxyquinoline-8-carboxylic acid methyl ester
Compound 159b (900 mg,4.8 mmol) was dissolved in AcOH (5 mL) and H was then added 2 O 2 (30%, 1 mL), and reacted at 85℃for 4 hours. Pouring the reaction solution into NaHCO 3 In solution, DCM was extracted, saturated NaCl solution was washed with water and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated under reduced pressure and the resulting residue was purified by column chromatography (MeOH: dcm=5%) to give methyl 3-hydroxyquinoline-8-carboxylate 159c (200 mg, light brown solid), yield: 21%. MS m/z (ESI): 204.2[ M+1 ] ] + .
Third step
Preparation of 3-methoxyquinoline-8-carboxylic acid methyl ester
Compound 159c (200 mg,1 mmol), K 2 CO 3 (408 mg,3 mmol) and CH 3 I (210 mg,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. The filtrate was filtered and concentrated under reduced pressure and the resulting residue was purified by column chromatography (EtOAc: pe=30%) to give methyl 3-methoxyquinoline-8-carboxylate 159d (200 mg, orange solid), yield: 89%.
MS m/z(ESI)：218.2[M+1] + .
Fourth step
Preparation of (3-methoxyquinolin-8-yl) methanol
Compound 159d (250 mg,0.92 mmol) was dissolved in dry THF (10 mL) and LiAlH4 (218 mg,5.75 mmol) was added at 0deg.C and reacted for 20min at 0deg.C. The reaction solution was saturated with NH 4 The Cl solution was quenched, extracted with EtOAc, washed with saturated NaCl and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated under reduced pressure and 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: 55%.
MS m/z(ESI)：190.2[M+1] + .
Fifth step
Preparation of 8- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -3-methoxyquinoline
Compound 159e (125 mg,0.66 mmol), 4-fluoro-2-methoxy-5-nitrophenol (136 mg,0.73 mmol) and PPh 3 (225 mg,0.86 mmol) was dissolved in dry THF (10 mL) and DIAD (174 mg,0.86 mmol) 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, pale yellow solid), yield: 40%.
MS m/z(ESI)：359.1[M+1] + .
Sixth step
Preparation of 2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) aniline
Compound 159f (90 mg,0.25 mmol) was dissolved in MeOH (10 mL) and saturated NH 4 Cl (2 mL) mixtureTo the combined solution, iron powder (140 mg,2.5 mmol) was then added. The reaction was carried out at room temperature for 2 hours, filtered, and the filtrate was concentrated 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 afforded 159g (70 mg, light brown solid) of 2-fluoro-4-methoxy-5- ((3-methoxyquinolin-8-yl) methoxy) aniline, yield: 68%.
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 (70 mg,0.21 mmol) of the compound and TEA (65 mg,0.64 mmol) were dissolved in anhydrous THF (5 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (107 mg,0.32 mmol) and reaction at room temperature for 16h. The reaction solution 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%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Compound 159H (60 mg,0.11 mmol) was dissolved in THF/MeOH/H 2 O=3:1:1 (10 mL) and then LiOH (13 mg,0.53 mmol) was added and reacted at room temperature for 1h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by 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%.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid (Cpd-074)
First step
Preparation of 3- (bromomethyl) benzothiophene
3-methylbenzothiophene 160a (200 mg,1.35 mmol), N-bromosuccinimide (288 mg,1.62 mmol) and azobisisobutyronitrile (110 mg,0.67 mmol) were dissolved in carbon tetrachloride (5 mL) followed by stirring at 85℃for 2 hours under nitrogen atmosphere. After the reaction was completed, 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 crude product obtained was purified by silica gel column (petroleum ether) to give 3- (bromomethyl) benzothiophene 160b (100 mg, white solid), yield: 32%.
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
Preparation of 3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene
3- (bromomethyl) benzothiophene 160b (100 mg,0.39 mmol), 4-fluoro-2-methoxy-5-nitrophenol (250 mg,1.40 mmol), potassium carbonate (1011 mg,5.4 mmol) and potassium iodide (52 mg,0.21 mmol) were dissolved in acetonitrile (5 mL). The mixture was stirred at 50℃for 2 hours. After the completion of the reaction, 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. Purification of the crude product on a silica gel column (ethyl acetate/petroleum ether=50%) afforded 3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene 160c (55 mg, white solid), yield: 40%.
MS m/z(ESI)：356.1[M+1] + 。
Third step
Preparation of 5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyaniline
3- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzothiophene 160c (55 mg,0.15 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of iron powder (650 mg,9.4 mmol) and saturated ammonium chloride solution (5 mL). The reaction was stirred at room temperature 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 (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%. The crude product was used directly in the next reaction.
MS m/z(ESI)：304.1[M+1] + 。
Fourth step
Preparation of dimethyl 4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
5- (benzothiophen-3-ylmethoxy) -2-fluoro-4-methoxyaniline 160d (50 mg,0.13 mmol) was dissolved in tetrahydrofuran (6 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (686 mg,2.0 mmol) and triethylamine (162 mg,1.52 mmol) 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 dimethyl 4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 160e (65 mg, yellow solid), yield: 82%. The crude product was used directly in the next reaction.
MS m/z(ESI)：545[M+1] + 。
Fifth step
Preparation of 3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- (benzothien-3-ylmethoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 160e (65 mg,0.11 mmol) was dissolved in tetrahydrofuran/water=1: 1 (5 mL) and then lithium hydroxide (108 mg,2.6 mmol) was added. The reaction was carried out at room temperature Stirred for 2 hours. After the completion of the reaction, 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. The crude product was purified by preparation (NH 3 ) To obtain 3- (5- (benzothiophene-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%.
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)
First step
Preparation of 1-benzoyl-3- (4-fluoro-3-methylphenyl) thiourea
4-fluoro-3-methylaniline 163a (3 g,24 mmol) was dissolved in tetrahydrofuran (10 mL). Benzoyl isothiocyanate (7.83 g,48 mmol) was then added at 25 ℃. The reaction solution was reacted at 25℃for 16 hours. Then spin-dry the solvent followed by ethanol: water = 4:1 (50 mL) was filtered 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
Preparation of (4-fluoro-3-methylphenyl) thiourea
1-benzoyl-3- (4-fluoro-3-methylphenyl) thiourea 163b (8.00 g,27.7 mmol) was dissolved in methanol: water = 3:1 (60 mL). Potassium carbonate (11.49 g,83.1 mmol) was then added at 25 ℃. The reaction solution was reacted at 70℃for 2 hours. Then spin-drying the solvent, adding water and filtering to obtain filter residue to obtain (4-fluoro-3-methylphenyl) thiourea 163c (4 g, white solid) with the yield of 79%.
MS m/z(ESI)：185[M+1] + 。
Third step
Preparation of 6-fluoro-7-methyl-1, 3-benzothiazol-2-amine
(4-fluoro-3-methylphenyl) thiourea 163c (4.00 g,21 mmol) was dissolved in chloroform (50 mL), and then liquid bromine (4.16 g,23 mmol) was added dropwise at 25 ℃. The reaction solution was reacted at 60℃for 2 hours. The solvent was then spun dry and the residue was filtered with water to give a mixture of 6-fluoro-7-methyl-1, 3-benzothiazol-2-amine 163d and 6-fluoro-5-methylbenzo [ d ] thiazol-2-amine 170d (2 g,163d:170 d=2:3, white solid) in 45% yield, as the mixture could not be separated by column and the peak time of the high performance liquid chromatography was essentially the same, the following steps were taken together and treated, the fourth to ninth steps were not specifically described for the synthesis step of molecule Cpd-076, and Cpd-075 synthesis steps were taken as examples until the last step was prepared and could not be separated.
MS m/z(ESI)：183[M+1] + 。
Fourth step
Preparation of 6-fluoro-7-methyl-1, 3-benzothiazole
6-fluoro-7-methyl-1, 3-benzothiazol-2-amine 163d (1.8 g,0.01 mol) was dissolved in 1, 4-dioxane (20 mL). Tert-butyl nitrite (3.06 g,0.30 mol) was then added, 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] + 。
Fifth step
Preparation of 5- (bromomethyl) -6, 8-difluoroquinoxaline
6-fluoro-7-methyl-1, 3-benzothiazole 163e (650 mg,3.6 mmol) was dissolved in carbon tetrachloride (15 mL). Dibenzoyl peroxide (436.mg, 1.8 mmol) was then added at 25℃with N-bromosuccinimide [706mg,3.96mmol ], and the reaction was allowed to react 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] + 。
Sixth step
Preparation of 6-fluoro-7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole
5- (bromomethyl) -6, 8-difluoroquinoxaline 163f (600 mg,2.44 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (458 mg,2.43 mmol) were dissolved in acetonitrile (10 mL). Then, potassium carbonate (673 mg,4.87 mmol) and potassium iodide (603 mg,3.66 mmol) were added, and the reaction mixture was reacted at 65℃for 2 hours. The solvent was then dried by spinning 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 (500 mg,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 (79 mg,1.4 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was extracted with dichloromethane (2X 10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, 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-dicarboxylic acid
2-fluoro-5- [ (6-fluoro-1, 3-benzothiazol-7-yl) methoxy ] -4-methoxyaniline 163h (450 mg,1.4 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (4638 mg,1.39 mmol) were dissolved in tetrahydrofuran (10 mL.) then triethylamine (423 mg,4.20 mmol) was added and the reaction mixture was reacted at 25℃for 1 hour.
MS m/z(ESI)：564.0[M+1] + 。
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-dicarboxylic acid 163i (10 mL stock solution) was dissolved in methanol with lithium hydroxide (21 mg,0.078 mmol): water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=5:95) to give 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) in 15% yield.
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-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-076 (6.21 mg, white solid) in 32% yield.
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)
First step
Preparation of 1-benzoyl-3- (3-methylphenyl) thiourea
3-methylaniline 165a (5 g,0.05 mol) was dissolved in tetrahydrofuran (80 mL). Benzoyl isothiocyanate (15 g,0.09 mol) was then added at 25 ℃. The reaction solution was reacted at 25℃for 16 hours. Then spin-dry the solvent followed by ethanol: water = 4: filtration of 1 (50 mL) gave after removal of the residue 1-benzoyl-3- (3-methylphenyl) thiourea 165b (14 g, yellow solid) in 88% yield.
MS m/z(ESI)：271.1[M+1] + 。
Second step
Preparation of (3-methylphenyl) thiourea
1-benzoyl-3- (3-methylphenyl) thiourea 165b (5 g,0.02 mol) was dissolved in methanol: water = 3:1 (60 mL). Potassium carbonate (7.67 g,0.05 mol) was then added at 25 ℃. The reaction solution was reacted at 70℃for 2 hours. The solvent was then spin-dried, and the residue was filtered with water to give (3-methylphenyl) thiourea 165c (2.5 g, white solid) in 64% yield.
MS m/z(ESI)：167.1[M+1] + 。
Third step
Preparation of 7-methyl-1, 3-benzothiazol-2-amine
(3-methylphenyl) thiourea 165c (1.8 g,0.01 mol) was dissolved in chloroform (50 mL), and then liquid bromine (2.1 g,0.01 mol) was added dropwise at 25 ℃. The reaction solution was reacted at 60℃for 2 hours. The solvent was then spun dry and the residue was filtered with water to give a mixture of 7-methyl-1, 3-benzothiazol-2-amine 165d and 5-methyl-1, 3-benzothiazol-2-amine 171d (1 g,165d: 171d=1:3, white solid) in 45% yield, the mixture was not isolated by column and the peak time of the high performance liquid chromatography was essentially the same, the following steps were taken together and treated together, the fourth to ninth steps of the synthesis of molecule Cpd-078 were not specifically described again, and Cpd-077 synthesis was taken as an example until the last step was prepared and could not be isolated.
MS m/z(ESI)：165.1[M+1] + 。
Fourth step
Preparation of 7-methyl-1, 3-benzothiazole
7-methyl-1, 3-benzothiazol-2-amine 165d (1 g, 0.006mol) was dissolved in 1, 4-dioxane (20 mL). Tert-butyl nitrite (1.9 g,0.018 mol) was then added and the reaction mixture 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] + 。
Fifth step
Preparation of 7- (bromomethyl) -1, 3-benzothiazole
7-methyl-1, 3-benzothiazole 165e (600 mg,4.02mmol ]) was dissolved in carbon tetrachloride (15 mL). NBS (858 mg,4.82 mmol) and BPO (487 mg,2.01 mmol) were then added at 25℃and the reaction was allowed to react 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] + 。
Sixth step
Preparation of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole
7- (bromomethyl) -1, 3-benzothiazole 165f (800 mg,3.50 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (787 mg,4.20 mmol) were dissolved in acetonitrile (30 mL). Then, potassium carbonate (969 mg,7.01 mmol) and potassium iodide (873 mg,5.26 mmol) were added, and the reaction mixture was reacted at 65℃for 2 hours. The solvent was then dried by spinning and extracted with dichloromethane (2X 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and the concentrated residue was purified by column chromatography (petroleum ether/ethyl acetate: 70/30) to give 165g (600 mg, yellow solid) of 7- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole 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
165g (300 mg,0.89 mmol) of 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 (1 g,17 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, and the filtrate was 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] + 。
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 (200 mg,0.65 mmol) and dimethyl 4- [ ((phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (550 mg,1.64 mmol) were dissolved in tetrahydrofuran (10 mL.) then triethylamine (268 mg,6.57 mmol) was added and the reaction mixture was reacted at 25℃for 16 hours.
MS m/z(ESI)：564.0[M+1] + 。
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-dicarboxylic acid 165i (10 mL stock solution) was dissolved in methanol with lithium hydroxide (61 mg,1.28 mmol): water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 90 minutes. After the reaction was completed, it was directly concentrated and preliminarily purified by reverse column (water: acetonitrile=5:95) to give 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) in a yield of 20%.
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) in 65% yield.
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)
First step
Preparation of 4-chloro-2-fluoro-6-nitroaniline
2-fluoro-6-nitroaniline 166a (2 g,12.8 mmol) and N-chlorosuccinimide (1.71 g,12.8 mmol) were dissolved in N, N-dimethylformamide (20 mL). The reaction solution was stirred at 80℃for 16 hours under nitrogen protection. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=8%) gave 4-chloro-2-fluoro-6-nitroaniline 166b (2.13 g, yellow solid), yield: 86%.
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
Preparation of 2-bromo-5-chloro-1-fluoro-3-nitrobenzene
Cuprous bromide (2.82 g,19.6 mmol) and t-butyl nitrite (4.05 g,39.3 mmol) were dissolved in acetonitrile (10 mL) and a solution of 4-chloro-2-fluoro-6-nitroaniline 166b (2.5 g,13.1 mmol) in acetonitrile (10 mL) was added dropwise at 50deg.C. 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=5%) gave 2-bromo-5-chloro-1-fluoro-3-nitrobenzene 166c (2.5 g, yellow solid), yield: 73%.
1 H NMR(400MHz,CDCl 3 )δ7.68(t,J＝2.0Hz,1H),7.40(dd,J＝7.6,2.4Hz,1H).
Third step
Preparation of 2-bromo-5-chloro-3-fluoroaniline
2-bromo-5-chloro-1-fluoro-3-nitrobenzene 166c (2.5 g,9.8 mmol), iron powder (1.64 g,29.4 mmol) and ammonium chloride (1.57 g,29.4 mmol) were dissolved in ethanol: water = 1:1 (30 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 (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), yield: 77%. The crude product was used directly in the next reaction.
MS m/z(ESI)：223.9[M+1] + 。
Fourth step
Preparation of 1-benzoyl-3- (2-bromo-5-chloro-3-fluorophenyl) thiourea
2-bromo-5-chloro-3-fluoroaniline 166d (2 g,8.9 mmol) was dissolved in tetrahydrofuran (30 mL), and benzoyl isothiocyanate (2.9 g,17.8 mmol) was added dropwise to the solution. The reaction was stirred at 25℃for 16 hours under air protection. After the reaction was completed, 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%.
MS m/z(ESI)：387.0(M+H)。
Fifth step
Preparation of (2-bromo-5-chloro-3-fluorophenyl) thiourea
1-benzoyl-3- (2-bromo-5-chloro-3-fluorophenyl) thiourea 166e (3.4 g,8.8 mmol) and potassium carbonate (2.43 g,17.6 mmol) were dissolved in methanol: water=4:1 (20 mL), and the reaction was stirred at 70℃for 2 hours. After the reaction was completed, 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). The filter cake was (2-bromo-5-chloro-3-fluorophenyl) thiourea 166f (1.58 g, white solid), yield: 62%. MS m/z (ESI): 283.0[ M+1 ]] + 。
Sixth step
Preparation of 4-bromo-7-chloro-5-fluoro-1, 3-benzothiazol-2-amine
(2-bromo-5-chloro-3-fluorophenyl) thiourea 166f (1.58 g,5.6 mmol) was dissolved in chloroform (20 mL) and bromine (1.07 g,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 was added an aqueous solution of sodium thiosulfate (50 mL). 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%.
MS m/z(ESI)：281.0[M+1] + 。
Seventh step
Preparation of 4-bromo-7-chloro-5-fluoro-1, 3-benzothiazole
166g (1.8 g,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 (1.98 g,19.2 mmol) was added dropwise to the solution at room temperature. The mixture was stirred at 90℃for 1 hour. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=6%) gave 4-bromo-7-chloro-5-fluoro-1, 3-benzothiazole 166h (1.05 g, yellow solid), yield: 61%.
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 methyl 7-chloro-5-fluoro-1, 3-benzothiazole-4-carboxylate
4-bromo-7-chloro-5-fluoro-1, 3-benzothiazole 166h (1.05 g,3.9 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1.43 g,1.97 mmol) and triethylamine (954 mg,7.88 mmol) were dissolved in methanol (20 mL) and then the reaction was stirred at 100℃under carbon monoxide (1.5 MPa) for 24 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=15%) gave methyl 7-chloro-5-fluoro-1, 3-benzothiazole-4-carboxylate 166i (174 mg, white solid), yield: 18%.
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).
Ninth step
Preparation of (7-chloro-5-fluoro-1, 3-benzothiazol-4-yl) methanol
Methyl 7-chloro-5-fluoro-1, 3-benzothiazole-4-carboxylate 166i (150 mg,0.61 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL) followed by dropwise addition of diisobutylaluminum hydride solution (1.0M in n-hexane) (1.2 mL) to the solution at 0deg.C. The mixture was stirred at 0℃for 2 hours. After the reaction was completed, the reaction solution was quenched with saturated potassium sodium 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%. The crude product was used directly in the next reaction.
MS m/z(ESI)：218.0[M+1] + 。
Tenth step
Preparation of 4- (bromomethyl) -7-chloro-5-fluoro-1, 3-benzothiazole
(7-chloro-5-fluoro-1, 3-benzothiazol-4-yl) methanol 166j (130 mg,0.60 mmol) was dissolved in dichloromethane (10 mL), and phosphorus tribromide (161 mg,0.60 mmol) was added dropwise to the solution at 0deg.C. The mixture was stirred at 0℃for 1 hour. After the completion of the reaction, the reaction mixture was quenched with water (10 mL), extracted with methylene chloride (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration gave the crude 4- (bromomethyl) -7-chloro-5-fluoro-1, 3-benzothiazole 166k (110 mg, yellow solid), yield: 60%. The crude product was used directly in the next reaction.
MS m/z(ESI)：280.0[M+1] + 。
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 (110 mg,0.39 mmol)), 4-fluoro-2-methoxy-5-nitroaniline (88 mg,0.47 mmol), potassium carbonate (108 mg,0.78 mmol) and potassium iodide (32 mg,0.19 mmol) were dissolved in acetonitrile (10 mL). The mixture was stirred at 50℃for 2 hours. After the 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. Purification of the crude product by silica gel column (ethyl acetate/petroleum ether=23%) gave 7-chloro-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole 166l (65 mg, white solid), yield: 42%.
MS m/z(ESI)：387.0[M+1] + 。
Twelfth step
Preparation of 5- [ (7-chloro-5-fluoro-1, 3-benzothiazol-4-yl) methoxy ] -2-fluoro-4-methoxyaniline
7-chloro-5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -1, 3-benzothiazole 166l (60 mg,0.15 mmol), iron powder (43 mg,0.77 mmol) and ammonium chloride (41 mg,0.77 mmol) were dissolved 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 (10 mL), ethyl acetate (3×10 mL) was extracted, 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), yield: 88%. 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 (45 mg,0.13 mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (50 mg,0.15 mmol) and triethylamine (25 mg,0.25 mmol) to the solution. The resulting mixture was stirred at room temperature for 16 hours. After completion of the reaction, 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) was obtained after filtration and concentration in yield: 53%. 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
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 (45 mg,0.075 mmol) was dissolved in tetrahydrofuran/water=1: 1 (5 mL) and then lithium hydroxide (10 mg,0.37 mmol) was added. The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture 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 preparation of (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%.
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)
First step
Preparation of diethyl 2, 3-dicyanofumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL,0.07 mol) and ethyl 2-cyanoacetate 004a (5 mL,0.04 mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction solution was stirred at 90℃under reflux for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid was precipitated. The reaction solution was filtered and washed with ice-ethanol to give diethyl 2, 3-dicyanofumarate 004b (2.33 g, white solid), yield: 26%.
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
Preparation of 5-amino-1H-pyrazole-3, 4-dicarboxylic acid ethyl ester
Diethyl 2, 3-dicyanofumarate 004b (500 mg,2.25 mmol) was dissolved in ethanol (10 mL), and acethydrazide (218 mg,2.48 mmol) and sodium acetate (18 mg,0.23 mmol) were added to react the reaction solution at 90℃for 0.5 hours. After the reaction was completed, it was cooled to room temperature and extracted with methylene chloride (3X 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: 50% ethyl acetate/50% petroleum ether) to give ethyl 5-amino-1H-pyrazole-3, 4-dicarboxylate 004c (333 mg, white solid), yield: 31%.
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).
Third step
Preparation of ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxylate
Ethyl 5-amino-1H-pyrazole-3, 4-carboxylate 004c (390 mg,1.24 mmol) was dissolved in methylene chloride (4 mL), 1' -carbonyldiimidazole (241 mg,1.49 mmol), triethylamine (376 mg,3.72 mmol) and 5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyaniline (282 mg,1.24 mmol) were added, and the reaction solution was reacted at 25℃for 1.5 hours. After completion of the reaction, water quenching was added, the organic phase was extracted with dichloromethane (3X 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated and purified by column chromatography (mobile phase: 5% methanol/95% dichloromethane) to give ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxylate 004d (554 mg, white solid) in 79% yield.
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).
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
Ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxy-late 004d (100 mg,0.18 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (38 mg,0.90 mmol) was added and the reaction was stirred at 25 ℃ for 4 hours after completion of the reaction, the reaction system was adjusted to be acidic with 2N diluted hydrochloric acid, dichloromethane was added to extract (2 x 20 mL) the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated and purified via a 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-4, 5,6, 7-tetrahydro-1H-pyrazolo [3,4-d ] pyrimidine-3-carboxylate Cpd-080 (17 mg, white solid) in 14% 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-dioxo-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-081)
First step
Preparation of 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4, 6-dioxo-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester
5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4, 6-dioxo-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 073a (100 mg,0.19 mmol) was dissolved in acetonitrile (10 mL) followed by the addition of methyl iodide (27 mg,0.19 mmol) and potassium carbonate (80 mg,0.58 mmol). Stirring was carried out at room temperature for 1h. After completion of the reaction, 6N hydrochloric acid solution was added to adjust the pH to 2-3, followed by purification by reverse column (acetonitrile/water: 75/25) to give the objective product 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4, 6-dioxo-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 073b (15 mg, white solid), yield: 10%.
MS m/z(ESI)：535.1[M+1] + 。
Third step
Preparation of 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4, 6-dioxo-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 073b (15 mg,0.03 mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL), followed by addition of lithium hydroxide monohydrate (4 mg,0.09 mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction, the solvent was removed by concentration under reduced pressure, followed by purification by reverse column (acetonitrile/water) and purification by preparation (acetonitrile/0.1% aqueous formic acid) to give 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1-methyl-4, 6-dioxo-7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid Cpd-081 (5 mg, white solid), yield: 27%.
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)
First step
Preparation of diethyl 2, 3-dicyanofumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL,0.07 mol) and ethyl 2-cyanoacetate 075a (5 mL,0.04 mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction solution was stirred at 90℃under reflux for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid was precipitated. The reaction solution was filtered and washed with ice-ethanol to give diethyl 2, 3-dicyanofumarate 075b (2.33 g, white solid), yield: 26%.
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
Preparation of 5-amino-1H-pyrazole-3, 4-dicarboxylic acid ethyl ester
Diethyl 2, 3-dicyanofumarate 075b (500 mg,2.25 mmol) was dissolved in ethanol (10 mL), and acethydrazide (218 mg,2.48 mmol) and sodium acetate (18 mg,0.23 mmol) were added to react the reaction solution at 90℃for 0.5 hours. After the reaction was completed, it was cooled to room temperature and extracted with methylene chloride (3X 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: 50% ethyl acetate/50% petroleum ether) to give 5-amino-1H-pyrazole-3, 4-dicarboxylic acid ethyl ester 075c (333 mg, white solid), yield: 31%.
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).
Third step
Preparation of ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxylate
Ethyl 5-amino-1H-pyrazole-3, 4-carboxylate 075c (390 mg,1.24 mmol) was dissolved in dichloromethane (4 mL), 1' -carbonyldiimidazole (241 mg,1.49 mmol), triethylamine (376 mg,3.72 mmol) and 5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyaniline (282 mg,1.24 mmol) were added, and the reaction solution was reacted at 25℃for 1.5 hours. After completion of the reaction, water quenching was added, the organic phase was extracted with dichloromethane (3X 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated and purified by column chromatography (mobile phase: 5% methanol/95% dichloromethane) to give ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxylate 075d (554 mg, white solid) in 79% yield.
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).
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
Ethyl 3- (3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) -1H-pyrazole-4, 5-dicarboxylate 075d (307 mg,0.54 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (65 mg,2.71 mmol) was added and the reaction was stirred at 25℃for 4 hours after completion of the reaction, the reaction system was adjusted to acidity with 2N dilute 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- (((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 075e (58 mg, white solid) in 22% yield.
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).
Fifth 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 methyl ester
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 (58 mg,0.12 mmol) was dissolved in dichloromethane (2 mL), oxalyl chloride (150 mg,1.18 mmol) and a catalytic amount of N, N-dimethylformamide were added, and reacted at 25℃for 10 minutes.
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-dioxo-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-083)
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First step
Preparation of diethyl 5- ({ [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] carbamoyl } amino) -1H-pyrazole-3, 4-dicarboxylic acid
Diethyl 5-amino-1H-pyrazole-3, 4-carboxylate (114 mg,0.50 mmol) was dissolved in dichloromethane (4 mL), 1' -carbonyldiimidazole (98 mg,0.60 mmol), triethylamine (153 mg,1.51 mmol) and 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyaniline 101a (150 mg,0.50 mmol) were added, and the reaction solution was reacted at 25℃for 1.5 hours. After the reaction was completed, water was added to quench, the organic phase was extracted with dichloromethane (3×30 mL), dried over anhydrous sodium sulfate and filtered, and 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, pale yellow solid), yield: 49%.
MS m/z(ESI)：552.1[M+1] + 。
Second step
Preparation of 5- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -4, 6-dioxo-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-carboxylate 101b (70 mg,0.13 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (30 mg,1.27 mmol) was added, and the reaction was stirred at 25℃for 4 hours. After completion of the reaction, the solution was concentrated and purified by a preparative column (mobile phase: 0.1% acetic acid/acetonitrile/water) to give Cpd-083 (5 mg, white solid), yield of 5- [ 2-fluoro-5- (isoquinolin-8-ylmethoxy) -4-methoxyphenyl ] -4, 6-dioxo-1 h,7 h-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid: 8%.
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-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid (Cpd-084)
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First step
Preparation of 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4, 6-dioxo-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester
Diethyl 5- [ ({ 5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } carbamoyl) amino ] -1H-pyrazole-3, 4-dicarboxylic acid 118a (200 mg,0.35 mmol) was dissolved in tetrahydrofuran, followed by addition of 4A molecular sieves (50 mg), stirring at room temperature for 30 minutes, and then sodium hydride (85 mg,3.53 mmol) was added under ice-bath conditions. The reaction was stirred at room temperature for 3 hours. After LCMS detection of completion of the reaction, sodium hydride was quenched by addition of saturated ammonium chloride solution under ice bath, followed by extraction 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 a silica gel column (petroleum ether/ethyl acetate=3/7) to give 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4, 6-dioxo-1 h,7 h-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 118b (120 mg, white solid), yield: 80%.
MS m/z(ESI)：521.1[M+1] + 。
Second step
Preparation of 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester
5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -4, 6-dioxo-1H, 7H-pyrazoline [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 073a (100 mg,0.19 mmol) was dissolved in acetonitrile (10 mL) followed by the addition of methyl iodide (27 mg,0.19 mmol) and potassium carbonate (80 mg,0.58 mmol). Stirring was carried out at room temperature for 1h. After completion of the reaction, 6N hydrochloric acid solution was added to adjust the pH to 2-3, followed by purification by reverse column (acetonitrile/water: 75/25) to give the objective product, ethyl 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylate 118c (50 mg, white solid), yield: 45%.
MS m/z(ESI)：549.4[M+1] + 。
Third step
Preparation of 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid
5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid ethyl ester 118c (50 mg,0.09 mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL), followed by addition of lithium hydroxide monohydrate (11 mg,0.27 mmol). The reaction was stirred at room temperature for 1h. After completion of the reaction, the solvent was removed by concentration under reduced pressure, followed by purification by reverse column (acetonitrile/water) and purification by preparation (acetonitrile/0.1% aqueous formic acid) to give 5- {5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-methoxyphenyl } -1, 7-dimethyl-4, 6-dioxapyrazolo [3,4-d ] pyrimidine-3-carboxylic acid Cpd-084 (30 mg, white solid), yield: 62%.
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-dione isoindole-4-carboxylic acid (Cpd-085)
First step
Preparation of 2- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1, 3-dione isoindole-4-carboxylic acid
Phenyl-1, 2, 3-tricarboxylic acid (70 mg,0.33 mmol) was dissolved in toluene (5 mL), and triethylamine (33.7 mg,0.33 mmol) and 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 (83.48 mg,27 mmol) 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 Cpd-085 (12 mg, brown solid) of 2- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1, 3-dione isoindole-4-carboxylic acid, yield: 9%.
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)
First step
Preparation of 4-fluoro-5-iodo-2-methoxyphenol
Methyl 4-fluoro-5-iodo-2-methoxybenzoate 024a (500 mg,1.6 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the sequential addition of methanol (2.5 mL), 1M sodium hydroxide solution (2.5 mL). The reaction solution 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%.
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
Preparation of 1, 2-difluoro-3- ((4-fluoro-5-iodo-2-methoxyphenoxy) methyl) -4-methoxybenzene
4-fluoro-5-iodo-2-methoxyphenol 024b (400 mg,1.49 mmol) was dissolved in N, N-dimethylformamide (20 mL), followed by the sequential addition of 2- (chloromethyl) -3, 4-difluoro-1-methoxybenzene (287 mg,1.49 mmol), potassium carbonate (412 mg,2.98 mmol), potassium iodide (124 mg,0.74 mmol). The reaction mixture was reacted at 80℃for 2 hours. Then extracted with ethyl acetate (2X 50 mL). The organic phases were combined, 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), yield: 69%.
MS m/z(ESI)：447.0(M+23)。
Third step
Preparation of 2- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan
1, 2-difluoro-3- ((4-fluoro-5-iodo-2-methoxyphenoxy) methyl) -4-methoxybenzene 024c (500 mg,1.20 mmol) was dissolved in dioxane (5 mL). Then, bis-pinacolato diboron (450 mg,1.75 mmol), potassium acetate (345 mg,3.55 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (85 mg,0.10 mmol) were added sequentially. The reaction was allowed to react 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, 5-tetramethyl-1, 3, 2-dioxaborolan 024d (200 mg, white solid), yield: 36%.
MS m/z(ESI)：424.2[M+1] + 。
Fourth step
Preparation of (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxybenzene) boronic acid
2- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan 024d (150 mg,0.35 mmol) was dissolved in acetone (5 mL), followed by the sequential addition of water (5 mL), sodium periodate (227 mg,1.06 mmol), ammonium acetate (109 mg,1.41 mmol). The reaction mixture was reacted at 25℃for 16 hours. Then extracted with ethyl acetate (2X 10 mL). The organic phases were combined, 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), yield: 89%.
MS m/z(ESI)：343.1[M+1] + 。
Fifth step
Preparation of 1- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid methyl ester
(5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxybenzene) boronic acid 024e (50 mg,0.15 mmol) was dissolved in toluene (5 mL) followed by the sequential addition of 1H-indole-3-carboxylic acid methyl ester (52.3 mg,0.3 mmol), pyridine (94.9 mg,1.2 mmol) and 1M lithium bis trimethylsilylamide (0.3 mL). The reaction mixture was reacted at 100℃for 16 hours. The reaction solution was filtered off with suction, and the filtrate was concentrated and subjected to preparative HPLC to give methyl 1- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylate 024f (30 mg, yellow oil) in yield: 30%.
MS m/z(ESI)：473.3[M+1] + 。
Sixth step
Preparation of 1- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid
1- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid methyl ester 024f (30 mg,0.06 mmol) was dissolved in tetrahydrofuran (2 mL). Methanol (2 mL), water (1 mL), lithium hydroxide (2.87 mg,0.12 mmol) were then added sequentially. The reaction mixture was reacted at 25℃for 1 hour. After the reaction, the pH was adjusted to 2 to 3 with a 1M hydrochloric acid solution. The reaction solution was concentrated and purified by preparative HPLC to give Cpd-086 (11 mg, white solid), yield of 1- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) -1H-indole-3-carboxylic acid: 41%.
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).
Example 87 and example 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)
First step
Preparation of 2, 3-difluoro-6-methoxybenzaldehyde
2M lithium diisopropylamide solution (3.82 mL,7.64 mmol) was dissolved in tetrahydrofuran (10 mL), cooled to-78deg.C, 1, 2-difluoro-4-methoxybenzene 025a (1000 mg,6.94 mmol) in tetrahydrofuran (3 mL) was added dropwise, the reaction stirred at-78deg.C for 1 hour, then N, N-dimethylformamide (578 mg,7.64 mmol) was added dropwise at-78deg.C, and the reaction stirred at-78deg.C for 1 hour again. 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 saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by silica gel column (dichloromethane/methanol=30/1) to give the title product 2, 3-difluoro-6-methoxybenzaldehyde 025b (1050 mg, yellow solid), yield: 79%.
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
Preparation of 2, 3-difluoro-6-methoxybenzyl alcohol
2, 3-difluoro-6-methoxybenzaldehyde 025b (1000 mg,5.81 mmol) was dissolved in toluene (10 mL), and under ice-water bath, 1.1mL of 0.1N sodium hydroxide solution was added followed by sodium borohydride (264 mg,6.98 mmol). The reaction solution was stirred at 25℃for 4 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 filtrate was concentrated to give the title product 2, 3-difluoro-6-methoxybenzyl alcohol 025c (980 mg, yellow oil), yield: 87%.
MS m/z(ESI)：157.1[M+1] + .
Third step
Preparation of 2, 3-difluoro-6-methoxybenzyl chloride
2, 3-difluoro-6-methoxybenzyl alcohol 025c (480 mg,5.63 mmol) was dissolved in toluene (8 mL) and concentrated hydrochloric acid (4.7 mL) was added. The reaction solution was stirred at 50℃for 6 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 filtrate was concentrated to give the title product 2, 3-difluoro-6-methoxybenzyl chloride 025d (900 mg, yellow oil), yield: 75%.
Fourth step
Preparation of 2- (5-bromo-2-methoxyphenoxymethyl) -3, 4-difluoro-1-methoxybenzene
5-bromo-2-methoxyphenol (300 mg,1.48 mmol) was dissolved in N, N-dimethylformamide (5 mL), and 2, 3-difluoro-6-methoxybenzyl chloride 025d (428 mg,2.22 mmol), potassium carbonate (614 mg,4.44 mmol) and potassium iodide (369 mg,2.22 mmol) 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 saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the residue obtained by concentrating the filtrate was purified by 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%.
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).
Fifth step
Preparation of 1- {3- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid methyl ester
1H-indazole-6-carboxylic acid methyl ester (108 mg,0.62 mmol) was dissolved in N, N-dimethylacetamide (3 mL) under nitrogen atmosphere, 2- (5-bromo-2-methoxyphenoxymethyl) -3, 4-difluoro-1-methoxybenzene 025e (200 mg,0.56 mmol), cuprous iodide (I) (22 mg,0.11 mmol), trans-cyclohexane-1, 2-diamine (26 mg,0.22 mmol) and potassium phosphate (356 mg,1.68 mmol) 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 silica gel column (petroleum ether/ethyl acetate=1/1) to give the title product, 1- {3- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid methyl ester 025f (180 mg, pale yellow solid), yield: 70%.
MS m/z(ESI)：455.1[M+1] + .
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)
1- {3- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid methyl ester 025f (100 mg,0.22 mmol) was dissolved in a solution of methanol/water=4/1 (3 mL), lithium hydroxide monohydrate (47 mg,1.1 mmol) was added, and the reaction was stirred at 25℃for 2 hours. 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), which was 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%;2- {3- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -4-methoxyphenyl } indazole-6-carboxylic acid Cpd-088 (16.3 mg, pale yellow solid), yield: 16%.
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)
First step
Preparation of N- (3-fluoro-2-methylphenyl) acetamide
3-fluoro-2-methylaniline 133a (5.00 g,39.95 mmol) was dissolved in dichloromethane (30 mL), triethylamine (8.07 g,79.81 mmol) and acetyl chloride (3450 mg,43.95 mmol) were added at 0℃and stirred for 1 hour at 0 ℃. After the reaction was completed, water (20 mL) was added to quench, and extracted with DCM (3×50 mL), and the organic phase was washed with brine, dried over anhydrous sodium sulfate and filtered. FilteringThe solution was concentrated and dried to give N- (3-fluoro-2-methylphenyl) acetamide 133b (6.00 g, white solid), yield: 81%. MS m/z (ESI): 168.2[ M+1 ]] + 。
Second step
Preparation of N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide
N- (3-fluoro-2-methylphenyl) acetamide 133b (4.00 g,23.93 mmol) was dissolved in concentrated sulfuric acid (24 mL), fuming nitric acid (2.78 g,28.71 mmol) was added dropwise at 0deg.C, and then stirred at 0deg.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 give N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide 133c (3.00 g, yellow solid), yield: 56%.
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).
Third step
Preparation of 3-fluoro-2-methyl-6-nitroaniline
To a solution of N- (3-fluoro-2-methyl-6-nitrophenyl) acetamide 133c (3.00 g,14.08 mmol) in methanol (20 mL) was added dropwise concentrated sulfuric acid (20 mL) and stirred at 90℃for 4 hours. After the reaction was completed, the reaction system was concentrated, the pH was adjusted to 8-9 with 3M sodium hydroxide, then extracted with methylene chloride (3X 20 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and dried to give 3-fluoro-2-methyl-6-nitroaniline 133d (2.80 g, yellow solid), yield: 90%. MS m/z (ESI): 171.1[ M+1 ]] + 。
Fourth step
Preparation of 4-fluoro-3-methylbenzene-1, 2-diamine
To a solution of 3-fluoro-2-methyl-6-nitroaniline 133d (1.29 g,7.60 mmol) in ethanol (10 mL) were added saturated aqueous ammonium chloride (10 mL) and iron powder (1.27 g,22.80 mmol), and the mixture was stirred at 80℃for 1 hour. After the reaction, the mixture was filtered while it was still hot, the filtrate was extracted with methylene chloride (3X 30 mL), and the organic phase was dried over anhydrous sodium sulfateAfter filtration, the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=3/1) to give 4-fluoro-3-methylbenzene-1, 2-diamine 133e (517 mg, brown solid), yield: 49%. 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).
Fifth step
Preparation of 6-fluoro-5-methylquinoxaline
To a solution of 4-fluoro-3-methylbenzene-1, 2-diamine 133e (517 mg,3.69 mmol) in ethanol (10 mL) was added glyoxal (1.07 g,7.38mmol,40% aqueous solution), and the reaction was stirred at room temperature for 3 hours. After the reaction was completed, extracted with ethyl acetate (3×20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) to give 6-fluoro-5-methylquinoxaline 133f (160 mg, white solid), yield: 27%.
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).
Sixth step
Preparation of 5- (bromomethyl) -6-fluoroquinoxaline
To a solution of 6-fluoro-5-methylquinoxaline 133f (160 mg,0.99 mmol) in carbon tetrachloride (5 mL) were added 2,2' -azobis (2-methylpropanenitrile) (24 mg,0.10 mmol) and N-bromosuccinimide (176 mg,0.99 mmol), and the mixture was stirred at 80℃for 3 hours. After the reaction was completed, water was added to quench, extracted with methylene chloride (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to give 133g (208 mg, yellow solid) of 5- (bromomethyl) -6-fluoroquinoxaline, yield: 87%.
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 acetonitrile (10 mL) of 4-fluoro-2-methoxy-5-nitrophenol (178 mg,0.95 mmol) were added potassium carbonate (178 mg,1.29 mmol) and 133g (208 mg,0.86 mmol) of 5- (bromomethyl) -6-fluoroquinoxaline, and the reaction mixture was stirred at 70℃for 3 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, filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: petroleum ether/ethyl acetate=3/1) to give 6-fluoro-5- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) quinoxaline 133h (227 mg, white solid), yield: 76%.
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 (177 mg,0.51 mmol) in ethanol (3 mL) and water (3 mL) were added iron powder (85 mg,1.53 mmol) and ammonium chloride (82 mg,1.53 mmol) and the reaction mixture was stirred at 80 ℃ for 2 hours. After the reaction was completed, the filtrate was filtered while it is hot, extracted with methylene chloride (3×10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated and dried to give 2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyaniline 133i (155 mg, yellow solid), yield: 96%. MS m/z (ESI): 318.2[ 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).
Ninth step
Preparation of dimethyl 4- (3- (2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate to a solution of 2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyaniline 133i (125 mg,0.39 mmol) in tetrahydrofuran (5 mL) was added triethylamine (60 mg,0.59 mmol) and methyl 4- ((phenoxycarbonyl) amino) thiophene-dimethyl-2, 3-dicarboxylate (145 mg,0.43 mmol) and stirred at 25℃for 5 hours. After the reaction was completed, water was added to quench, extracted with dichloromethane (3×10 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 dimethyl 4- (3- (2-fluoro-5- ((6-fluoroquinoxalin-5-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 133j (148 mg, white solid), yield: 67%.
MS m/z(ESI)：559.1[M+1] + 。
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 (118 mg,0.21 mmol) in tetrahydrofuran (3 mL), methanol (3 mL) and water (3 mL) was added lithium hydroxide (15 mg,0.63 mmol), and the mixture was reacted at 25℃for 2 hours. After completion of the reaction, 2N 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 a 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) in 31% yield.
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)
First step
Preparation of 1, 2-difluoro-3-methyl-4-nitrobenzene
Compound 135a (10 g,78.1 mmol) was dissolved in concentrated sulfuric acid (50 mL), concentrated nitric acid (4.92 g,78.1 mmol) was slowly added dropwise at-10℃and reacted at 0℃for 1h. The reaction solution was poured into ice water and saturated NaHCO was used 3 The pH of the solution was adjusted to 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), yield: 74%.
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
Preparation of 3, 4-difluoro-2-methylaniline
Compound 135b (10 g,57.8 mmol) was dissolved in MeOH/NH 4 Cl=5:1 solution (100 mL), reduced iron powder (16.14 g,289 mmol) was added and reacted at room temperature for 2h. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the obtained 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 3, 4-difluoro-2-methylaniline 135c (6 g, yellow oily liquid), yield: 72.5%.
MS m/z(ESI)：144.2[M+1] + .
Third step
Preparation of N- (3, 4-difluoro-2-methylphenyl) acetamide
Compound 135c (6 g,42 mmol) and triethylamine (12.7 g,126 mmol) were dissolved in anhydrous DCM (60 mL) and acetyl chloride (3.9 g,50.4 mmol) was added dropwise under ice-bath and reacted at room temperature for 1h. The reaction mixture was quenched with water (50 mL), extracted with methylene chloride (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), yield: 42.9%.
MS m/z(ESI)：186.2[M+1] + .
Fourth step
Preparation of N- (3, 4-difluoro-2-methyl-6-nitrophenyl) acetamide
Compound 135d (1.5 g,8.1 mmol) was dissolved in concentrated sulfuric acid (30 mL), concentrated nitric acid (0.7 mL) was slowly added dropwise at-10℃and reacted at 0℃for 1h. The reaction solution was poured into ice water and saturated NaHCO was used 3 The pH of the solution was adjusted to 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), yield: 53.8%.
MS m/z(ESI)：231.0[M+1] + .
Fifth step
Preparation of 3, 4-difluoro-2-methyl-6-nitroaniline
Compound 135e (1 g,4.3 mmol) was dissolved in absolute ethanol (20 mL) and then concentrated sulfuric acid (3 mL) was added dropwise and reacted at 90℃for 4h. Concentrating under reduced pressure, dissolving the residue in water, and concentrating with saturated NaHCO 3 The pH of the solution was adjusted to 7, extracted with ethyl acetate (3X 50 mL), washed with saturated NaCl and dried over anhydrous sodium sulfate. Filtering, concentrating the filtrate under reduced pressure to obtain 3, 4-difluoro-2-methyl-6-nitroaniline 135f (800 mg, yellow solid), yield: 97.9%.
MS m/z(ESI)：189.1[M+1] + .
Sixth step
Preparation of 4, 5-difluoro-3-methylbenzene-1, 2-diamine
Compound 135f (2.3 g,12.2 mmol) was dissolved in MeOH/NH 4 To a solution (60 mL) of cl=5:1 was added reduced iron powder (6.81 g,122 mmol). The reaction was carried out at room temperature for 16h. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the obtained 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. Column chromatography of the residue obtained (EtOAc: pe=60%) purification gave 135g (1.17 g, yellow solid), yield: 61%.
MS m/z(ESI)：159.2[M+1] + .
Seventh step
Preparation of 6, 7-difluoro-5-methylquinoxaline
135g (500 mg,3.16 mmol) of the compound was dissolved in HF (10 mL), and an aqueous glyoxal solution (553mg, 9.48 mmol) was added dropwise at 0℃and reacted at 0℃for 1 hour. The reaction solution was poured into water and saturated NaHCO was used 3 The pH of the solution was adjusted to 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), yield: 26.4%.
MS m/z(ESI)：181.2[M+1] + .
Eighth step
Preparation of 5- (bromomethyl) -6, 7-difluoroquinoxaline
Compound 135h (150 mg,0.83 mmol) was dissolved in carbon tetrachloride (50 mL) and AIBN (13.4 mg,0.08 mmol) and NBS (148 mg,0.83 mmol) were added and reacted at 80℃for 2h. The reaction solution was poured into water, extracted with methylene chloride, 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), yield: 71.9%.
MS m/z(ESI)：259.1[M+1] + .
Ninth step
Preparation of 6, 7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) quinoxaline
Compound 135i (70 mg,0.27 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (76 mg,0.41 mmol) were dissolved in ACN (10 mL), and potassium carbonate (112 mg,0.81 mmol) 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%.
MS m/z(ESI)：366.0[M+1] + .
Tenth step
Preparation of 5- ((6, 7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyaniline
Compound 135j (80 mg,0.22 mmol) was dissolved in MeOH/nh4cl=5:1 solution (6 mL), reduced iron powder (122 mg,2.2 mmol) was added and reacted at room temperature for 1h. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the obtained 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 afforded 5- ((6, 7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 135k (65 mg, light brown solid), yield: 80%.
MS m/z(ESI)：336.1[M+1] + .
Eleventh step
Preparation of dimethyl 4- (3- (5- ((6, 7-difluoroquinoxalin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
Compound 135k (60 mg,0.18 mmol) and triethylamine (55 mg,0.54 mmol) were dissolved in dry THF (5 mL) and methyl 4- ((phenoxycarbonyl) amino) thiophene dimethyl-2, 3-dicarboxylate (90 mg,0.27 mmol) was added and reacted at room temperature for 16h. The reaction solution 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, pale yellow solid), yield: 69%.
MS m/z(ESI)：577.2[M+1] + .
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
Compound 135l was dissolved in (75 mg,0.13 mmol) in THF/H 2 To a mixed solution of o=3:1 (8 mL), lithium hydroxide (16 mg,0.65 mmol) was added and reacted at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC (ACN: H) 2 O (0.2% FA) =55%) to obtain 3- (5- ((6, 7-difluoro quinoxalin-5-yl) methoxy group) -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%.
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 (1 h,3 h) -dione
First step
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 (1 g,2 mmol) was dissolved in N, N-dimethylformamide (10 mL). N, O-dimethylhydroxylamine hydrochloride (230 mg,2.4 mmol) was then added at 25 ℃; propylphosphoric anhydride (1.91 g,6 mmol); n, N-diisopropylethylamine (260 mg,2 mmol). The reaction mixture was reacted at 90℃for 1 hour. Then repeatedly extracted three times with water (50 mL) and dichloromethane (50 mL), 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
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 (1000 mg,1.8 mmol) was dissolved in tetrahydrofuran (10 mL). Methyl magnesium bromide (320 mg,2.7 mmol) was then added at 0deg.C. The reaction solution was reacted at 0℃for 2 hours. After the reaction was completed, it was directly concentrated and preliminarily purified by reverse column (water: acetonitrile=5:95) to give 5-acetyl-3- (5- (((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4-methoxyphenyl) thieno [3,4-d ] pyrimidine-2, 4 (1 h,3 h) -dione Cpd-094 (300 mg, yellow solid), yield: 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
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 (200 mg,0.3865 mmol) was dissolved in N, N-dimethylformamide (10 mL), and 1-propylphosphoric acid cyclic anhydride (368.93 mg,1.1595 mmol), N, N-diisopropylethylamine (149.85 mg,1.1595 mmol), methylamine hydrochloride (31.32 mg, 0.46338 mmol) were added sequentially at room temperature. The reaction was carried out at 90℃for 2 hours under nitrogen protection. After the reaction, the mixture was concentrated. Then extracted with ethyl acetate and concentrated, the concentrate was purified by a 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%
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
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
Oxalyl chloride (150 mg,1.18 mmol) and a catalytic amount 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-dioxo-1, 2,3, 4-tetrahydrothieno [3,4-d ] pyrimidine-5-carboxylic acid 177a (200 mg,0.39 mmol) in dichloromethane (5 mL) and the reaction was stirred at 25℃for 0.5 h. After completion of the TLC monitoring reaction, the reaction solution was concentrated. The concentrate was dissolved in methylene chloride (5 mL), triethylamine (60 mg,0.59 mmol) and methylamine hydrochloride (32 mg,0.47 mmol) were added, and the reaction was stirred at 25℃for 1 hour. After LCMS monitored completion of the reaction, quench with water, extract with dichloromethane (3×20 mL) and filter the organic phase after drying over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography 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%.
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-carboxylic acid sodium salt
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 (200 mg,0.39 mmol) in water (10 mL) was added sodium hydroxide (15 mg,0.39 mmol) and the reaction was stirred at 25℃for 0.5 hours. The reaction solution was lyophilized to give 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 Cpd-098 (200 mg, white solid), yield: 96%.
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
Preparation of choline salt 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-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 (200 mg,0.39 mmol) in water (10 mL) was added choline hydroxide (47 mg,0.39 mmol) and the reaction was stirred at 25 ℃ for 0.5 hours. 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-carboxylic acid choline salt Cpd-099 (232 mg, white solid), yield: 97%.
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-dioxo-1, 2,3, 4-tetrahydrothiophene (3, 4-d) pyrimidine-5-carboxylic acid
First step
Preparation of 6-ethoxy-2, 3-difluorobenzyl alcohol
6-ethoxy-2, 3-difluorobenzaldehyde 181a (420 mg,2.26 mmol) was dissolved in methanol (6 mL) and sodium borohydride (428.7 mg,11.3 mmol) was added portionwise. The reaction solution was stirred at 80℃for 4 hours. After the reaction was completed, the reaction was quenched by addition of a small amount of water (2 mL), dried by spin, and the crude product was purified by silica gel column (methanol/dichloromethane=8%) to give (6-ethoxy-2, 3-difluorophenyl) methanol 181b (280 mg, colorless oil), yield: 62.0%.
MS m/z(ESI)：171.1(M+H-18)。
Second step
Preparation of 2- (bromomethyl) -1-ethoxy-3, 4-difluorobenzene
(6-ethoxy-2, 3-difluorophenyl) methanol 181b (350 mg,1.86 mmol), tetrabromomethane (925.2 mg,2.8 mmol) and triphenylphosphine (731.8 mg,2.8 mmol) were dissolved in dichloromethane (10 mL). The reaction was stirred at 25℃for 3 hours under hydrogen protection. After the reaction was completed. The reaction system 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), 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).
Third step
Preparation of N- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (bromomethyl) -1-ethoxy-3, 4-difluorobenzene 181c (300 mg,1.19 mmol), 4-fluoro-2-methoxy-5-nitroaniline (266.9 mg,1.43 mmol) and potassium carbonate (329.8 mg,2.39 mmol) were dissolved in N, N-dimethylformamide (4 mL) and stirred at 80℃for 2h under nitrogen protection. After completion of the reaction, water (40 mL) was added to the reaction system to quench, the mixture was extracted with EA (3X 5 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by 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%.
MS m/z(ESI)：357.1(M+H)。
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 (400 mg,1.12 mmol) was dissolved in ethanol: water=4:1 (10 mL) and ammonium chloride (328.4 mg,5.61 mmol) iron powder (314.4 mg,5.61 mmol) was added the reaction mixture was stirred at 80℃for 2 hours after completion of the reaction, the filter cake was filtered, washed with ethanol, concentrated and the crude product 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)。
Fifth step
Preparation of dimethyl 4- (3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2, 3-dicarboxylate
1-N- ((6-ethoxy-2, 3-difluorophenyl) methyl) -4-fluoro-6-methoxybenzene-1, 3-diamine (150 mg,0.46 mmol), 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid methyl ester (154 mg,0.46 mmol) was dissolved in THF (6 mL), and triethylamine (278.6 mg,2.76 mmol) was added dropwise to the system. The reaction was carried out at 25℃for 8 hours. After the reaction was completed, the crude product was concentrated and purified by silica gel chromatography (pure ethyl acetate) to give dimethyl 4- (3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2, 3-dicarboxylate 181f (200 mg, yellow solid), yield: 72.1%.
MS m/z(ESI)：568.2(M+H)。
Sixth step
Preparation of 3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1, 2,3, 4-tetrahydrothiophene (3, 4-d) pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2, 3-dicarboxylic acid 181f (150 mg,0.26 mmol) lithium hydroxide monohydrate (22.18 mg 0.53 mmol) was dissolved in methanol: tetrahydrofuran: in water=1:1:1 (3 mL), the reaction mixture was stirred at 25 ℃ for 2 hours. After completion of the reaction, the crude product was purified by silica gel chromatography (dichloromethane: methanol=10:1) to give Cpd-100 (114 mg, yellow solid), yield of 3- (5- ((6-ethoxy-2, 3-difluorophenyl) methyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1H-thiophene (3, 4-d) pyrimidine-5-carboxylic acid: 82.7%.
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
First step
Preparation of 1, 2-difluoro-4-isopropoxy benzene
3, 4-difluorophenol 182a (5 g,38.4 mmol) was dissolved in acetone (50 mL), 2-iodopropane (7.83 g,46.1 mmol) and potassium carbonate (10.61 g,76.8 mmol) were added to the solution, the reaction solution was stirred overnight at 56℃and after the 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-isopropoxyphenyl 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
Preparation of 2, 3-difluoro-6-isopropoxybenzaldehyde
1, 2-difluoro-4-isopropoxybenzene 182b (3.9 g,22.7 mmol) was dissolved in tetrahydrofuran (20 mL), a 2M solution of lithium diisopropylamide in tetrahydrofuran (13 mL,24.5 mmol) was slowly added dropwise at-78deg.C, the reaction was stirred at this temperature for half an hour, then DMF (3.3 g,45.4 mmol) was added and stirring continued for half an hour. After the reaction, acetic acid and water are added for quenching reaction respectively, ethyl acetate extraction is carried out, organic phase is dried and concentrated, and then silica gel column (petroleum ether/ethyl acetate=10/1) is used for separation and purification to obtain 2, 3-difluoro-6-isopropoxybenzaldehyde 182c (850 mg, colorless oily liquid) with the 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).
Third step
Preparation of (2, 3-difluoro-6-isopropoxyphenyl) methanol
2, 3-difluoro-6-isopropoxybenzaldehyde 182c (850 mg,4.25 mmol) was dissolved in methanol (10 mL), sodium borohydride (322 mg,8.5 mmol) was added at 0 ℃ and 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 to quench, extracted with ethyl acetate, washed with aqueous sodium chloride, and after organic phase drying and concentration, separated and purified by a silica gel column (petroleum ether/ethyl acetate=10/1) to give (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).
Fourth step
Preparation of 2- (bromomethyl) -3, 4-difluoro-1-isopropoxy benzene
(2, 3-difluoro-6-isopropoxyphenyl) methanol 182d (680 mg,3.36 mmol) was dissolved in methylene chloride (20 mL), triphenylphosphine (1.06 g,4.03 mmol) was added at 0℃and stirred for 10 minutes, then carbon tetrabromide (1.34 g,4.03 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was concentrated and purified by a silica gel column (petroleum ether/ethyl acetate=10/1) to give 2- (bromomethyl) -3, 4-difluoro-1-isopropoxy benzene 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).
Fifth step
Preparation of N- (2, 3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline (382 mg,2.05 mmol) was dissolved in DMF (5 mL), 2- (bromomethyl) -3, 4-difluoro-1-isopropoxybenzene 182e (600 mg,2.26 mmol) was further added, potassium carbonate (567 mg,4.1 mmol) was reacted at 80℃for 2 hours, after the reaction was completed, water was added to the reaction solution, ethyl acetate was extracted, and after drying and concentration, N- (2, 3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 182f (340 mg, yellow solid) was isolated and purified by silica gel column (petroleum ether/ethyl acetate=5/1) in 44.8% yield.
MS m/z(ESI)：371.1(M+1)。
Sixth step
N 1 Preparation of- (2, 3-difluoro-6-isopropoxybenzyl) -4-fluoro-6-methoxyphenyl-1, 3-diamine
N- (2, 3-difluoro-6-isopropoxybenzyl) -4-fluoro-2-methoxy-5-nitroaniline 182f (340 mg,0.918 mmol) was dissolved in ethanol/saturated aqueous solution of ammonium chloride=4/1 (10 mL), iron powder (514 mg,9.18 mmol) was added, the reaction mixture was stirred at 80℃for 2 hours, after the reaction was completed, the reaction mixture was filtered by suction, the filtrate was extracted with ethyl acetate, and the organic phase was concentrated by drying and then separated and purified by a silica gel column (petroleum ether/ethyl acetate=10/1) to give 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 (190 mg, 0.5538 mmol) and 4- [ ((phenoxycarbonyl) amino)]Thiophene-2, 3-dicarboxylic acid methyl ester (225 mg,0.67 mmol) was dissolved in tetrahydrofuranTo the reaction mixture was added triethylamine (169 mg,1.67 mmol) and the mixture was stirred at room temperature for 16 hours, after the completion of the reaction, the mixture was concentrated and purified by separation on a silica gel column (petroleum ether/ethyl acetate=3/1) to give dimethyl 4- (3- (5- ((2, 3-difluoro-6-isopropylbenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate (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
Dimethyl 4- (3- (5- ((2, 3-difluoro-6-isopropylbenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 182h (270 mg, 0.460 mmol) was dissolved in tetrahydrofuran/water=1/1 (10 mL), lithium hydroxide (22 mg,0.928 mmol) was added thereto, the reaction mixture was stirred at room temperature for 1 hour, after completion of the reaction, the reaction mixture was concentrated to obtain 3- (5- ((2, 3-difluoro-6-isopropyloxybenzyl) 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) after preparation. 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-dioxo-1, 2,3, 4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid
First step
Preparation of 4- (cyclopropylmethoxy) -1, 2-difluorobenzene
3, 4-difluorophenol 184a (1.30 g,10.00 mmol) was dissolved in N, N-dimethylformamide (5 mL), bromomethylcyclopropane (1.35 g,10.00 mmol), potassium carbonate (4.15 g,30.00 mmol) and potassium iodide (1.66 g,10.00 mmol) were added sequentially, and the reaction solution was reacted at 90℃for 16 hours. After completion of the reaction, water (10 mL) was added, and the mixture was extracted with ethyl acetate (3X 10 mL) and washed with saturated brine (2X 10 mL). The combined organic phases were 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
Preparation of 6- (cyclopropylmethoxy) -2, 3-difluorobenzaldehyde
4- (cyclopropylmethoxy) -1, 2-difluorobenzene 184b (690 mg,3.75 mmol) was dissolved in ultra-dry tetrahydrofuran (10.0 mL). After stirring the reaction flask at-78 ℃ for 0.5 hours, a solution of lithium diisopropylamide in tetrahydrofuran (3.7 mL,2 mol/L) was slowly added over 15 minutes, the reaction solution was stirred at-78 ℃ for 20 minutes, N-dimethylformamide (301 mg,4.12 mmol) was slowly added dropwise, the mixture was allowed to react at-78 ℃ for further 0.5 hour, acetic acid (0.2 mL), water (1.5 mL) was slowly added dropwise after the reaction was completed, then extracted with ethyl acetate (3 x10 mL), and then the organic phase was combined, dried over anhydrous sodium sulfate and 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 184c (623 mg, colorless oily liquid) in a yield of 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).
Third step
Preparation of (6- (cyclopropylmethoxy) -2, 3-difluorophenyl) methanol
6- (Cyclopropylmethoxy) -2, 3-difluorobenzaldehyde 184c (623 mg,2.94 mmol) was dissolved in methanol (10 mL), and sodium borohydride (222 mg,5.87 mmol) was added under ice water bath. The reaction solution was reacted at 0℃for 20 minutes. After the reaction was completed, saturated ammonium chloride was added to quench it, followed by dilution with ethyl acetate (15 mL), washing with water (5 mL) and saturated brine (2×5 mL) in this order, and 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).
Fourth step
Preparation of 2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene
(6- (Cyclopropylmethoxy) -2, 3-difluorophenyl) methanol 184d (442 mg,2.06 mmol) was dissolved in methylene chloride (4 mL), triphenylphosphine (649 mg,2.48 mmol) and carbon tetrabromide (823mg, 2.48 mmol) were sequentially added under an ice-water bath, and the reaction solution was reacted at 0℃for 1 hour. After the reaction was completed, water (2 mL) was added, and the mixture was extracted with methylene chloride (3X 10 mL) and washed with saturated brine (2X 10 mL). The combined organic phases were 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).
Fifth step
Preparation of N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene 184e (400 mg,1.44 mmol) was dissolved in acetonitrile (10 mL), 4-fluoro-2-methoxy-5-nitroaniline (322 mg,1.73 mmol), potassium carbonate (599 mg,4.33 mmol) and potassium iodide (240 mg,1.44 mmol) were added sequentially, and the reaction solution was reacted at 65℃for 2 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 combined organic phases were 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).
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 (100 mg,0.26 mmol) was dissolved in tetrahydrofuran/H 2 O (2.5 ml, V/v=4:1). Ammonium chloride (42 mg,0.78 mmol) and iron powder (44 mg,0.78 mmol) were then added and the reaction was allowed to react at 25℃for 16 hours. Then filtered, the filtrate was extracted with dichloromethane (2×10 mL). The organic phases are combined, dried with 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 (95 mg,0.27 m)mol) and 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (90 mg,0.27 mmol) were dissolved in tetrahydrofuran (2 mL). Triethylamine (82 mg,0.81 mmol) was then added and the reaction was allowed to react at 25℃for 2 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: 75/25) to give dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) urea) thiophene-2, 3-dicarboxylate 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-dioxo-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-dicarboxylic acid 184h (107 mg,0.18 mmol) and lithium hydroxide (13 mg,0.54 mmol) were dissolved in tetrahydrofuran: water=4:1 (2.5 mL). The reaction solution was reacted at room temperature for 2 hours. After the reaction was completed, the mixture was directly filtered and concentrated and purified by a preparative reverse column (water: acetonitrile=20:80) to give 3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1, 2,3, 4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid Cpd-102 (15.56 mg, yellow solid) in a yield of 15.75%.
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
First step
Preparation of 4-fluoro-2-methoxy-5-nitrophenol 188b
To 4-fluoro-2-methoxy-5-nitrophenol 188a (2 g,0.0107 mol) was added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120℃for 12h. After the reaction was completed, 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), yield: 59.81%.
MS m/z(ESI)：174.1(M+1).
Second step
Preparation of 2- (2, 3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 188c
4-fluoro-2-methoxy-5-nitrophenol 188b (100 mg,0.5777 mmol) was dissolved in 20mL DMF, naH (27.73 mg,1.1554 mmol) was added at 0deg.C and stirred at that temperature for 10 min, then 2-bromo-3, 4-difluoro-1-methoxybenzene (128.84 mg,0.5777 mmol) was added and reacted at 25deg.C for 1 h after stirring for 10 min. After the reaction was completed, the reaction system was adjusted to acidity with 2N diluted hydrochloric acid, and ethyl acetate was added for extraction (2×20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 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%.
MS m/z(ESI)：330.1(M+1)。
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 (100 mg,0.30 mmol) was dissolved in methanol (5 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (169.62 mg,3.04 mmol) were added, and the reaction mixture was reacted at 25℃for 1 hour. Then filtered, the filtrate was extracted with dichloromethane (2×10 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated to give 4-amino-2- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -5-fluorophenol 188d (80 mg, yellow solid), yield: 79.22%.
MS m/z(ESI)：300.1(M+1)。
Fourth step
Preparation of methyl 4- [ ((5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2, 3-dicarboxylate 188e
4-amino-2- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -5-fluorophenol (80 mg,0.25 mmol) and methyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (158 mg,0.50 mmol) were dissolved in tetrahydrofuran (5 mL). Triethylamine (77 mg,0.75 mmol) was then added and the reaction was allowed to react at 25℃for 1 hour. The reaction solution was not subjected to any post-treatment to obtain crude methyl 4- [ ((5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2, 3-dicarboxylate 188e.
MS m/z(ESI)：541.1(M+1)。
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
Methyl 4- [ ((5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoro-4-hydroxyphenyl) carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 188e (5 mL stock solution) and lithium hydroxide (13 mg,0.53 mmol) were dissolved in methanol: water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 2 hours. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) 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 a yield of 87.46%.
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-dioxo-1, 2,3, 4-tetrahydrothiophene (3, 4-d) pyrimidine-5-carboxylic acid
First step
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 (2.5 g,7 mmol), formaldehyde (0.45 g,14 mmol) was dissolved in acetonitrile (30 mL) and stirred at 40℃under nitrogen for 0.5h. Sodium cyanoborohydride (0.66 g,10.5 mmol) and acetic acid (0.02 g,0.3 mmol) were added. The reaction mixture was stirred at 40℃for 4h. After the reaction was completed, 1Ml of water was added to quench, concentrate, dilute with water (30 Ml), extract with EA (20 Ml), concentrate the organic phase, and purify the crude product 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%.
MS m/z(ESI)：371.1(M+H)。
Second step
N 1 Preparation of- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-6-methoxy-N1-toluene-1, 3-diamine
N- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline (700 mg,1.89 mmol), ammonium chloride (553 mg,9.45 mmol) was dissolved in EtOH/H2O=4:1 (10 mL), and iron (529 mg,9.45 mmol) was added. The reaction mixture was stirred at 80℃for 2 hours. After the reaction is finished, filtering, concentrating filtrate, 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%.
MS m/z(ESI)：341.2(M+H)。
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 (400 mg,1.175 mmol), 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (390 mg,1.175 mmol) was dissolved in THF (20 mL), and triethylamine (719mg, 7.052 mmol) was added dropwise. The reaction mixture was stirred at 25℃for 1 hour. After the reaction was completed, the crude product was concentrated to give dimethyl 4- (3- (5- ((6-ethoxy-2, 3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate (500 mg, yellow solid) purified by silica gel column (dichloromethane: methanol=10:1), yield: 68.7%.
MS m/z(ESI)：582.1(M+H)。
Fourth step
Preparation of 3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1, 2,3, 4-tetrahydrothiophene (3, 4-d) pyrimidine-5-carboxylic acid
Dimethyl 4- (3- (5- ((6-ethoxy-2, 3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate (500 mg,0.26 mmol) lithium hydroxide monohydrate (22 mg 0.53 mmol) 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 completed, the crude product was concentrated and purified by silica gel chromatography (dichloromethane: methanol=10:1) to give 3- (5- ((6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) -2, 4-dioxo-1, 2,3, 4-tetrahydrothiophene (3, 4-d) pyrimidine-5-carboxylic acid Cpd-104 (40.4 mg, yellow solid), yield: 8.8%.
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
First step
Preparation of N-cyclopropyl-4-fluoro-2-methoxy-5-nitroaniline
1-bromo-4-fluoro-2-methoxy-5-nitrobenzene 190a (1 g, 0.004mol) was dissolved in toluene (20 mL), cyclopropylamine (0.27 g,0.0048 mol), dibenzylideneacetone dipalladium (0.37 g,0.0004 mol), 2- (dicyclohexylphosphine) -3, 6-dimethoxy-2 '-4' -6 '-tri-I-propyl-11' -biphenyl (0.64 g,0.0012 mol), sodium t-butoxide (0.58 g, 0.006mol) was added, and after the reaction was completed at 100℃for 16 hours under nitrogen gas, the temperature was recovered, and the mixture was filtered. Water (10 mL) was added, ethyl acetate (3X 10 mL) was used to extract, the organic phase was dried, concentrated, and the concentrate was further purified over a 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: 43%.
MS m/z(ESI)：227(M+1)。
Second step
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 (400 mg,1.5932 mmol) was dissolved in acetonitrile (10 mL), and 2- (bromomethyl) -1-ethoxy-3, 4-difluorobenzene (396.43 mg,1.7525 mmol), potassium carbonate (660.57 mg,4.7796 mmol), potassium iodide (26.45 mg,0.1593 mmol) was added. After the reaction was completed at 65℃for two hours, acetonitrile was removed by spinning, and water (10 mL) and ethyl acetate (3X 10 mL) were added to extract, 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%.
MS m/z(ESI)：397(M+1)。
Third step
Preparation of N-cyclopropyl-N- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1, 3-diamine
N-cyclopropyl-N- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 190c (300 mg,0.7569 mmol) was dissolved in acetonitrile (5 mL), iron powder (211.36 mg,3.7845 mmol) and ammonium chloride (121.46 mg,2.2707 mmol) were added, reacted at 80℃for one hour, after the reaction was completed, acetonitrile was spun off, water (10 mL) and ethyl acetate (3X 10 mL) were added to extract, and the organic phase was dried and concentrated. 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%.
MS m/z(ESI)：367(M+1)。
Fourth step
Preparation of dimethyl 4- (3- (5- (cyclopropyl (6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
N-cyclopropyl-N- (6-ethoxy-2, 3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1, 3-diamine 190d (220 mg,0.6005 mmol) was dissolved in tetrahydrofuran (5 mL) and 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (241.64 mg,0.7206 mmol), triethylamine (182.29 mg,1.8015 mmol) was added. After the reaction was completed, water (10 mL) and ethyl acetate (3×10 mL) were added thereto and extracted. The organic phase was dried and concentrated. The concentrate was further purified by silica gel column (petroleum ether/ethyl acetate=2/1) to give dimethyl 4- (3- (5- (cyclopropyl (6-ethoxy-2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 190e (150 mg, yellow solid). Yield: 40%.
MS m/z(ESI)：607(M+1)
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
To a solution of 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,0.34 mmol) in methanol/tetrahydrofuran/water=1/1/1 (3 mL) was added lithium hydroxide (41.39 mg, 1.528 mmol), and after the reaction was completed at room temperature for one hour, the solvent was dried by spin-drying, and the crude product was prepared by a preparative column (acetonitrile/water=1/4) to give 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 Cpd-105 (61 mg, yellow solid). Yield: 31%.
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
First step
Preparation of N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline
4-fluoro-2-methoxy-5-nitroaniline (1 g,5.37 mmol) was dissolved in methanol (20 mL), a few drops of acetic acid and 6- (cyclopropylmethoxy) -2, 3-difluorobenzaldehyde 191a (1.37 g,6.45 mmol) were added, the reaction mixture was stirred at room temperature for 15 minutes, then sodium cyanoborohydride (1.01 g,16.11 mmol) was added, the reaction mixture was stirred at 40 ℃ for 2 hours, after the completion of the reaction, quenched by addition of aqueous ammonium chloride solution, extracted with ethyl acetate, and after the organic phase was concentrated by dryness, N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 191b (1 g, yellow solid) was isolated and purified by a silica gel column (petroleum ether/ethyl acetate=3/1), the yield was 48.7%.
MS m/z(ESI)：383.1(M+1)。
Second step
Preparation of N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline
N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-5-nitroaniline 191b (1 g,2.61 mmol) was dissolved in methanol (20 mL), a few drops of acetic acid and aqueous formaldehyde were added, the reaction mixture was stirred at room temperature for 15 minutes, then sodium cyanoborohydride (493 mg,7.85 mmol) was added, the reaction mixture was stirred at 40℃for 16 hours, after the completion of the reaction, an aqueous ammonium chloride solution was added for quenching, extraction with ethyl acetate, and separation and purification by a silica gel column (petroleum ether/ethyl acetate=10/1) gave N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 191c (900 mg, yellow solid) in 87% yield.
MS m/z(ESI)：397.1(M+1)。
Third step
N 1 - (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-6-methoxy-N 1 Preparation of (E) -methylbenzene-1, 3-diamine
N- (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-2-methoxy-N-methyl-5-nitroaniline 191c (900 mg,2.27 mmol) was dissolved in ethanol/aqueous ammonium chloride=4/1 (25 mL), iron powder (1.27 g,22.7 mmol) was added, the reaction mixture was stirred at 80℃for 2 hours, after completion of the reaction, suction filtration was performed, and the filtrate was concentrated and purified by silica gel column (petroleum ether/ethyl acetate=10/1) to give N 1 - (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-6-methoxy-N 1 Toluene-1, 3-diamine 191d (430 mg, yellow solid) in 51.7% yield.
MS m/z(ESI)：367.2(M+1)。
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 (430 mg,1.17 mmol) was dissolved in tetrahydrofuran (10 mL) followed by the addition of 4- ((phenoxycarbonyl) amino) thiophene-dimethyl 2, 3-diformate (470 mg,1.41 mmol) and triethylamine (356 mg,3.52 mmol) were stirred at room temperature for 16 hours, after the reaction was completed, the reaction mixture was concentrated and purified by separation on a silica gel column (petroleum ether/ethyl acetate=3/1) to give dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-diformate 191e (600 mg, yellow solid) in 84.4% yield.
MS m/z(ESI)：608.2(M+1)。
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
Dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) (methyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 191e (600 mg,0.987 mmol) was dissolved in tetrahydrofuran/water=1/1 (10 mL), followed by addition of lithium hydroxide (71 mg,2.96 mmol), the reaction solution was stirred at room temperature for 1 hour, after the completion of the reaction, the reaction solution was concentrated to prepare 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
First step
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 (650 mg,2.87 mmol) and 2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene (876 mg,3.16 mmol) were dissolved in acetonitrile (30 mL), followed by the addition of potassium carbonate (792 mg,5.74 mmol) and potassium iodide (470 mg,2.87 mmol), and the reaction stirred at 70℃for 16 hours. After the reaction, water was added to the reaction mixture, extraction was performed with ethyl acetate, and after drying and concentration of the organic phase, the organic phase was 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) in 39.6% yield.
MS m/z(ESI)：423.1(M+1)。
Second step
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 (480 mg,1.14 mmol) was dissolved in ethanol/aqueous ammonium chloride=4/1 (25 mL), iron powder (345 mg,11.4 mmol) was then added, the reaction mixture was stirred at 80℃for 2 hours, after the completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated and purified by separation on a silica gel column (petroleum ether/ethyl acetate=5/1) to give 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)。
Third step
Preparation of dimethyl 4- (3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
Will N 1 -cyclopropyl-N 1 - (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) -4-fluoro-6-methoxybenzene-1, 3-diamine 192c (180 mg,0.459 mmol) was dissolved in tetrahydrofuran (10 mL), followed by addition of 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester (169 mg,0.504 mmol) and triethylamine (139 mg,1.37 mmol), stirring of the reaction solution at 40℃for 16 hours, concentrating the reaction solution after completion of the reaction, and separating and purifying the reaction solution by a silica gel column (petroleum ether/ethyl acetate=3/1) 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)。
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
Dimethyl 4- (3- (5- (cyclopropyl (6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) amino) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 192d (270 mg,0.426 mmol) was dissolved in tetrahydrofuran/water=1/1 (10 mL), followed by addition of lithium hydroxide (30 mg,1.28 mmol), and after the reaction was stirred at room temperature for 1 hour, the reaction mixture 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). Yield: 30%.
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
First step
Preparation of diethyl 2, 3-dicyanofumarate
Tetrahydrofuran (5 mL) was added dropwise to thionyl chloride (6 mL,0.07 mol) and ethyl 2-cyanoacetate 193a (5 mL,0.04 mol) was added slowly with stirring. After the completion of the dropwise addition, the reaction solution was stirred at 90℃under reflux for 3 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, and a solid was precipitated. The reaction solution was filtered and washed with ice-ethanol to give diethyl 2, 3-dicyanofumarate 193b (2.3 g, white solid), yield: 21.04%.
MS m/z(ESI)：223.1(M+1).
Second step
Preparation of 5-amino-1H-pyrazole-3, 4-dicarboxylic acid diethyl ester
Diethyl 2, 3-dicyanofumarate 193b (2 g,0.009 mol) was dissolved in ethanol (10 mL), and acetohydrazide (0.87 g,0.0099 mol) and sodium acetate (0.07 g,0.0009 mol) were added to the reaction mixture, which was reacted at 90℃for 0.5 hours. After the reaction was completed, it was cooled to room temperature and extracted with dichloromethane (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: 50% ethyl acetate/50% petroleum ether) to give diethyl 5-amino-1H-pyrazole-3, 4-dicarboxylate 193c (1.2 g, white solid), yield: 58.89%.
MS m/z(ESI)：228.1(M+1).
Third step
Preparation of diethyl 5- ([ (5- ([ 6- (cyclopropylmethoxy) -2, 3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3, 4-dicarboxylate
Diethyl 5-amino-1H-pyrazole-3, 4-carboxylate 193c (501.6 mg,2.21 mmol) was dissolved in dichloromethane (20 mL), 1' -carbonyldiimidazole (429.54 mg,2.65 mmol), triethylamine (670.13 mg,6.62 mmol) and 5- ([ 6- (cyclopropylmethoxy) -2, 3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyaniline (780 mg,2.21 mmol) were added, and the reaction solution was reacted at 25℃for 1.5 hours. After completion of the reaction, water quench was added, extraction with dichloromethane (3×30 mL), drying of the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate and purification by column chromatography (mobile phase: 5% methanol/95% dichloromethane) gave diethyl 5- ([ (5- ([ 6- (cyclopropylmethoxy) -2, 3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3, 4-dicarboxylate 193d (500 mg, white solid), yield: 37.3%.
MS m/z(ESI)：607.2(M+1).
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
Diethyl 5- ([ (5- ([ 6- (cyclopropylmethoxy) -2, 3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) carbamoyl ] amino) -2H-pyrazole-3, 4-dicarboxylic acid 193d (100 mg,0.18 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (38 mg,0.90 mmol) was added, and the reaction was stirred at 25℃for 4 hours. After the reaction was completed, the reaction system was adjusted to acidity with 2N diluted hydrochloric acid, and extracted with dichloromethane (2×20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 10% ethyl acetate/80% petroleum ether) to give 5- (5- ([ 6- (cyclopropylmethoxy) -2, 3-difluorophenyl ] methoxy) -2-fluoro-4-methoxyphenyl) -4, 6-dioxo-1 h,7 h-pyrazolo [3,4-d ] pyrimidine-3-carboxylic acid Cpd-108 (3.5 mg, white solid), yield: 3.82%.
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) -diones
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 (200 mg,1.57 mmol) 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-dioxo-1, 2,3, 4-tetrahydrothiophene [3,4-d ] pyrimidine-5-carboxylic acid 177a (400 mg,0.79 mmol) in dichloromethane (5 mL) and the reaction stirred at 25℃for 15 min. After completion of the reaction, the reaction mixture was concentrated, tetrahydrofuran (3 mL) was added to dissolve, then an aqueous solution in which sodium borohydride (60 mg,1.57 mmol) was dissolved was added dropwise, and after stirring at 0 ℃ for 15 minutes, water and ethyl acetate (3×10 mL) were added to extract, and the organic phase was dried over sodium sulfate, concentrated, and the concentrated solution 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 (1 h,3 h) -dione Cpd-109 (202.62 mg, white solid), yield: 52%.
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
First step
Preparation of tert-butyl (tert-butoxycarbonyl) (2-chloro-5-fluoropyridin-4-yl) carbamate
2-chloro-4-amino-5-fluoropyridine 196a (1 g,6.82 mmol) was dissolved in acetonitrile (20 mL), followed by addition of di-tert-butyl dicarbonate (3.72 g,17.05 mmol) and 4-dimethylaminopyridine (167 mg,1.36 mmol), stirring of the reaction solution at room temperature was carried out for 16 hours, after completion of the reaction, the reaction solution was concentrated and then separated and 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 42.3% yield.
MS m/z(ESI)：347.1(M+1)。
Second step
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 (1 g,2.88 mmol) and (6- (cyclopropylmethoxy) -2, 3-difluorophenyl) methanol (611 mg,2.88 mmol) were dissolved in 1, 4-dioxane (20 mL) followed by the addition of BrettPhos (309 mg,0.576 mmol), pd 2 (dba) 3 (264 mg, 0.284 mmol) and cesium carbonate (1.88 g,5.76 mmol), the reaction mixture was stirred at 80 ℃ for 2 hours, after the completion of the reaction, water was added, ethyl acetate was used for extraction, the organic phase was dried over anhydrous sodium sulfate, concentrated and purified by separation on 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)。
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 (400 mg,0.762 mmol) was dissolved in dichloromethane (10 mL), then HCl/1, 4-dioxane solution (5 mL) was added, the reaction solution was stirred at 35 ℃ for 16 hours, after the reaction was completed, the reaction solution was concentrated, saturated aqueous sodium bicarbonate solution was added, ethyl acetate was extracted, the organic phase was dried over anhydrous sodium sulfate, and after concentration, 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)。
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 (200 mg, 0.611 mmol) was dissolved in tetrahydrofuran (5 mL), followed by the addition of dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (627mg, 1.851 mmol) and triethylamine (187 mg,1.851 mmol), the reaction mixture was stirred at room temperature for 16 hours, after the reaction was completed, the reaction mixture was concentrated and purified by separation on a silica gel column (petroleum ether/ethyl acetate=2/1) to give dimethyl 4- (3- (2- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) ureido) thiophene-2, 3-dicarboxylate 196e (10 mg, yellow solid) in 2.9% yield.
MS m/z(ESI)：566.2(M+1)。
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
Dimethyl 4- (3- (2- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -5-fluoropyridin-4-yl) ureido) thiophene-2, 3-dicarboxylic acid 196e (10 mg,0.017 mmol) was dissolved in tetrahydrofuran/water=1/1 (2 mL), followed by addition of lithium hydroxide (2 mg,0.085 mmol), the reaction mixture was stirred at room temperature for 1 hour, and after completion of the reaction it was spun-dried to give 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) in 22.6% yield.
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
First step
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 (14 mg,0.026 mmol) was dissolved in dichloromethane (1 mL), DIPEA (10 mg,0.078 mmol) was added, msCl (3 mg,0.026 mmol) was added at 0deg.C, and the reaction stirred for 1 hour, then used directly in the next step without any treatment.
MS m/z(ESI)：549.1(M+1)。
Second step
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,0.052 mmol) was added to the reaction solution of the previous step, the reaction solution was stirred at room temperature for 1 hour, after the completion of the reaction, the reaction solution was concentrated to obtain 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
First step
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 (40 mg,0.075 mmol) and manganese dioxide (33 mg,0.37 mmol) 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 give a crude product 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, which is used directly in the next step.
MS m/z(ESI)：533.0(M+1)。
Second step
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-carbaldehyde 198b (35 mg,0.066 mmol) and dimethyl ammonia hydrochloride (11 mg,0.131 mmol) were dissolved in methanol (5 mL). The reaction was stirred at 40℃for 30 minutes, then sodium cyanoborohydride (21 mg,0.328 mmol) was added to the reaction solution, and the reaction solution was stirred at 40℃for 1 hour. After the reaction was completed, the crude product was purified by preparation to give 3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) -5- ((dimethylamino) methyl) thieno [2,3-d ] pyrimidine-2, 4 (1 h,3 h) -dione Cpd-112 (9.26 mg, white solid), yield: 22.53%.
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-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
First step
Preparation of 6-fluoro-2, 3-dimethoxybenzoic acid 199b
LDA (1.51 g,0.014 mol) was added dropwise to 4-fluoro-1, 2-dimethoxybenzene 199a (2 g,0.013 mol) under nitrogen atmosphere at-78deg.C, and stirred at-60deg.C for 30 minutes. The dried carbon dioxide was then added and stirring 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 extraction was performed with ethyl acetate. 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%.
MS m/z(ESI)：201.1(M+1).
Second step
Preparation of 6-fluoro-2, 3-dihydroxybenzoic acid 199c
To 6-fluoro-2, 3-dimethoxybenzoic acid 199b (1.2 g,0.0107 mol) was added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120℃for 12h. After the reaction was completed, 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%.
MS m/z(ESI)：173.1(M+1).
Third step
Preparation of methyl 6-fluoro-2, 3-dihydroxybenzoate 199d
6-fluoro-2, 3-dihydroxybenzoic acid 199c (0.8 g,0.0046 mol) was dissolved in 20mL of methanol followed by SOCl addition at 0deg.C 2 (1.64 g,0.0138 mmol) and the mixture solution was stirred at 70℃for 16h. After the reaction was completed, 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).
Fourth step
Preparation of 5-fluoro-2H-1, 3-benzodioxolane-4-carboxylic acid methyl ester 199e
Methyl 6-fluoro-2, 3-dihydroxybenzoate 199d (500 mg,2.69 mmol), diiodomethane (1.08 g,4.03 mmol) and cesium carbonate (1.31 g,4.03 mmol) were dissolved in DMF (10 mL) and the reaction was allowed to react at 110℃for 1.5 h. After the reaction was completed, 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 methyl 5-fluoro-2H-1, 3-benzodioxolane-4-carboxylate 199e (320 mg, yellow solid), yield: 54.11%.
MS m/z(ESI)：199.1(M+1).
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 (300 mg,1.514 mmol) was dissolved in 5mL THF followed by addition of 5mL LiAlH under ice water bath 4 The mixture solution was stirred at 25℃for 2h. After completion of the reaction, extraction with ethyl acetate (3X 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: 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%.
MS m/z(ESI)：153.1(M-18).
Sixth step
Preparation of 4- (bromomethyl) -5-fluoro-2H-1, 3-benzodioxan 199g
(5-fluoro-2H-1, 3-Benzodioxol-4-yl) methanol 199f (100 mg,0.5878 mmol) and PBr 3 (190.93 mg,0.7053 mmol) in 5mL DCM and the mixture solution stirred at 25℃for 1h. After completion of the reaction, extraction was performed 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 column chromatography (mobile phase: 30% ethyl acetate/70% petroleum ether) to give 199g of 4- (bromomethyl) -5-fluoro-2H-1, 3-benzodioxane (100 mg, white solid), yield: 52.57%.
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-benzodioxan 199H
4-fluoro-2-methoxy-5-nitrophenol (100 mg,0.5344 mmol) was dissolved in 20mL DMF, naH (25.65 mg,1.0688 mmol) was added at 0deg.C and stirred at that temperature for 10 min, then 199g (124.53 mg,0.5344 mmol) of 4- (bromomethyl) -5-fluoro-2H-1, 3-benzodioxan was added and stirred for 10 min before reacting at 25deg.C for 1H. After the reaction was completed, the reaction system was adjusted to acidity with 2N diluted hydrochloric acid, and ethyl acetate was added for extraction (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: 10% ethyl acetate/90% petroleum ether) to give 5-fluoro-4- (4-fluoro-2-methoxy-5-nitrophenoxymethyl) -2H-1, 3-benzodioxan 199H (50 mg, yellow solid), yield: 19.85%.
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-benzodioxan 199H (50 mg,0.1474 mmol) was dissolved in methanol (10 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (82.32 mg,1.474 mmol) were added, and the reaction mixture was reacted at 80℃for 1 hour. Then filtered, the filtrate was extracted with dichloromethane (3×20 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated to give 2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyaniline 199i (40 mg, yellow solid), yield: 78.97%.
MS m/z(ESI)：310.0(M+1)。
Ninth step
Preparation of methyl 4- [ ({ 2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 199j
2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyaniline 199i (40 mg,0.1293 mmol) and methyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (86.72 mg,0.2586 mmol) were dissolved in tetrahydrofuran (10 mL). Triethylamine (39.25 mg,0.3879 mmol) was then added and the reaction was allowed to react at 25℃for 16 hours. The reaction solution was not subjected to any post-treatment to obtain crude stock 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)。
Tenth step
Preparation of 3- (2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid
Dimethyl 4- [ ({ 2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl } carbamoyl) amino ] thiophene-2, 3-dicarboxylic acid 199j (5 mL stock solution) was dissolved in methanol with lithium hydroxide (6.52 mg,0.2724 mmol): water=3:1 (4 mL). The reaction mixture was reacted at 25℃for 2 hours. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) to give 3- (2-fluoro-5- [ (5-fluoro-2H-1, 3-benzodioxol-4-yl) methoxy ] -4-methoxyphenyl) -2, 4-dioxo-1H-thieno [3,4-d ] pyrimidine-5-carboxylic acid Cpd-113 (7.73 mg, white solid) in a yield of 16.19%.
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
First step
Preparation of 2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene
(6- (cyclopropylmethoxy) -2, 3-difluorophenyl) methanol 201a (12 g,0.056 mol) was dissolved in methylene chloride (100 mL), followed by addition of triphenylphosphine (17.63 g,0.067 mol), carbon tetrabromide (22.29 g,0.067 mol), stirring of the reaction solution at room temperature was carried out for 1 hour, after completion of the reaction, the reaction solution was concentrated and then separated and purified by a silica gel column (petroleum ether) to give 2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene 201b (9 g, white solid) in 58% yield.
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
Preparation of 1- (cyclopropylmethoxy) -3, 4-difluoro-2- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) benzene
2- (bromomethyl) -1- (cyclopropylmethoxy) -3, 4-difluorobenzene 201b (3 g, 0.0111 mol) and 4-fluoro-2-methoxy-5-nitrophenol (2.02 g, 0.0111 mol) were dissolved in acetonitrile (100 mL), followed by addition of potassium carbonate (4.48 g,0.032 mol), potassium iodide (1.79 g, 0.0111 mol), stirring of the reaction solution at 70 ℃ for 16 hours, addition of water to the reaction solution after completion of the reaction, extraction with ethyl acetate, drying concentration of the organic phase, and separation and purification 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)。
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 (1 g,2.61 mmol) was dissolved in ethanol/aqueous ammonium chloride=4/1 (50 mL), iron powder (1.46 g,26.1 mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after the reaction was completed, the reaction solution was filtered, and after the filtrate was concentrated, the filtrate was separated and purified by 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)。
Fourth step
Preparation of dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline 201d (850 mg,2.41 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (887 mg,2.65 mmol) and triethylamine (730 mg,7.22 mmol), the reaction mixture was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction mixture was concentrated and purified by separation on a silica gel column (petroleum ether/ethyl acetate=3/1) to give dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 201e (1.3 g, yellow solid) in 91% yield.
MS m/z(ESI)：595.1(M+1)。
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
Dimethyl 4- (3- (5- ((6- (cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 201e (1.3 g,2.18 mmol) was dissolved in tetrahydrofuran/water=1/1 (20 mL), lithium hydroxide (0.16 g,6.54 mmol) was added, the reaction mixture was stirred at room temperature for 1 hour, tetrahydrofuran was selected after the reaction was completed, the pH was adjusted to be acidic with dilute hydrochloric acid, 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-114 (900 mg, yellow solid) in 75% yield.
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
First step
Preparation of 2-aminothiophene-3, 4-dicarboxylic acid ethyl ester
Ethyl 2-cyanoacetate 202a (10 g,88.4 mmol), ethyl 2-oxopropionate (10.26 g,88.4 mmol) and sulfur (2.83 g,88.6 mmol) were dissolved in N, N-dimethylformamide (50 mL), and triethylamine (9.84 g,97.2 mmol) was added to the solution under ice-bath conditions. The reaction solution was stirred at 50℃for 3 hours. After the reaction was completed, the reaction mixture 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. Purification of the crude product by silica gel column (petroleum ether/ethyl acetate=28%) gave 2-aminothiophene-3, 4-dicarboxylic acid ethyl ester 202b (16 g, yellow solid), yield: 73%.
MS m/z(ESI)：244.0(M+1)。
Second step
Preparation of diethyl 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3, 4-dicarboxylate
Phenyl p-nitrochloroformate (4.96 g,24.6 mmol) and sodium bicarbonate (2.07 g,24.6 mmol) were dissolved in acetonitrile (20 mL), and diethyl 2-aminothiophene-3, 4-dicarboxylate 202b (3 g,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 is finished, the reaction solution is filtered and concentrated to obtain a crude product of diethyl 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3, 4-dicarboxylate 202c, which is directly used in the next step without treatment.
MS m/z(ESI)：409.0(M+1)。
Third step
Preparation of diethyl 2- (3- (5- ((6-cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-3, 4-dicarboxylate
The crude product, diethyl 2- (((4-nitrophenoxy) carbonyl) amino) thiophene-3, 4-dicarboxylate 202c (1.89 g,4.6 mmol), 5- ((6-cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyaniline (1.5 g,4.2 mmol) and triethylamine (850 mg,8.4 mmol) were dissolved in tetrahydrofuran (20 mL). The reaction solution was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3×20 mL), and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by silica gel column (petroleum ether/ethyl acetate=23%) to give diethyl 2- (3- (5- ((6-cyclopropylmethoxy) -2, 3-difluorobenzyl) oxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-3, 4-dicarboxylate 202d (2.7 g, yellow solid), yield: 85%.
MS m/z(ESI)：623.2(M+1)。
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 (300 mg,0.48 mmol) was dissolved in tetrahydrofuran (5 mL) and then sodium hydride (35 mg,1.44 mmol) was added under ice-bath. The reaction was stirred under ice bath for 7 hours. After completion of the reaction, the reaction mixture 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 preparation (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%.
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
First step
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) propanoic acid2, 4-dioxa-1, 2,3, 4-tetrahydrothieno [2,3-d]Pyrimidine-5-carboxylic acid 203a (160 mg,0.29 mmol) was dissolved in tetrahydrofuran (5 mL), and a solution of lithium aluminum hydride in tetrahydrofuran (0.3 mL,0.29 mmol) was added to the solution under ice-bath conditions. The reaction solution was stirred at 0℃for 1 hour. After the reaction was completed, the reaction mixture was quenched with methanol (20 mL), 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 preparation (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 (1 h,3 h) -dione Cpd-116 (6.38 mg, white solid), yield: 4.01%.
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
First step
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 (300 mg,0.55 mmol) was dissolved in tetrahydrofuran (5 mL), 2M lithium aluminum hydride solution in tetrahydrofuran (0.55 mL,1.1 mmol) was slowly added dropwise at 0deg.C, the reaction stirred at room temperature for 2 hours, quenched with aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was spun dry to give 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
First step
Preparation of 4-fluoro-5-nitrobenzene-1, 2-diol 205b
To 4-fluoro-2-methoxy-5-nitrophenol 205a (2 g,0.0107 mol) was added 25mL of ACOH and 15mL of HBr, and the mixture solution was stirred at 120℃for 12h. After the reaction was completed, 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-5-nitrobenzene-1, 2-diol 205b (1.1 g, yellow solid), yield: 59.81%.
MS m/z(ESI)：174.1(M+1).
Second step
Preparation of 2- (2, 3-difluoro-6-methoxyphenoxy) -5-fluoro-4-nitrophenol 205c
4-fluoro-5-nitrobenzene-1, 2-diol 205b (100 mg,0.5777 mmol) was dissolved in 20mL DMF, naH (27.73 mg,1.1554 mmol) was added at 0deg.C and stirred at that temperature for 10 min, then 2-bromo-3, 4-difluoro-1-methoxybenzene (128.84 mg,0.5777 mmol) was added and reacted at 25deg.C for 1 h after stirring for 10 min. After the reaction was completed, the reaction system was adjusted to acidity with 2N diluted hydrochloric acid, and ethyl acetate was added for extraction (2×20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated and purified by column chromatography (mobile phase: 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%.
MS m/z(ESI)：330.1(M+1)。
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 (100 mg,0.3035 mmol), (bromomethyl) cyclopropane (40.95 mg,0.3035 mmol), potassium carbonate (5.05 mg, 0.003mmol) was dissolved in acetonitrile (10 mL) and stirred at 80℃for 2 hours. After the reaction was completed, 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 purified by 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), yield: 77.43%.
MS m/z(ESI)：384.2(M+1)。
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 (100 mg,0.2609 mmol) was dissolved in methanol (10 mL). Then, a saturated ammonium chloride solution (1 mL) and iron powder (145.71 mg, 2.319 mmol) were added, and the reaction mixture was reacted at 80℃for 1 hour. Then filtered, the filtrate was extracted with dichloromethane (3×20 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated to give 4- (cyclopropylmethoxy) -5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoroaniline 205e (100 mg, yellow solid), yield: 97.62%.
MS m/z(ESI)：354.1(M+1)。
Fifth step
Preparation of dimethyl 4- ({ [4- (cyclopropylmethoxy) -5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylate 205f
4- (Cyclopropylmethoxy) -5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluoroaniline 205e (100 mg,0.283 mmol) and dimethyl 4- [ (phenoxycarbonyl) amino ] thiophene-2, 3-dicarboxylate (189.8 mg,0.566 mmol) were dissolved in tetrahydrofuran (10 mL). Triethylamine (85.91 mg,0.849 mmol) was then added, and the reaction was reacted at 25℃for 16 hours. The reaction solution was not subjected to any post-treatment to obtain crude stock solution of dimethyl 4- ({ [4- (cyclopropylmethoxy) -5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 205f.
MS m/z(ESI)：595.1(M+1)。
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
Dimethyl 4- ({ [4- (cyclopropylmethoxy) -5- [ (2, 3-difluoro-6-methoxyphenyl) methoxy ] -2-fluorophenyl ] carbamoyl } amino) thiophene-2, 3-dicarboxylic acid 205f (5 mL stock solution) was dissolved in methanol with lithium hydroxide (12.08 mg,0.5046 mmol): water = 3:1 (4 mL). The reaction mixture was reacted at 25℃for 2 hours. After the reaction was completed, it was directly concentrated and purified by reverse column (water: acetonitrile=20:80) 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
First step
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 (150 mg,0.4556 mmol) was dissolved in acetonitrile (10 mL), 1-bromobutane 206b (74.91 mg, 0.552 mmol), potassium carbonate (125.94 mg,0.9112 mmol), potassium iodide (7.56 mg,0.04556 mmol) was added. After the reaction was completed at 90℃for 2 hours, the reaction was allowed to resume at room temperature. Spin-drying acetonitrile, and passing the residue 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
Preparation of 4-butoxy-5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline
To a mixture of 2- ((2-butoxy-4-fluoro-5-nitrophenoxy) methyl) -3, 4-difluoro-1-methoxybenzene 206c (150 mg,0.3893 mmol) in ethanol/water=1/1 (10 mL) was added iron powder (43.48 mg,0.7786 mmol) and ammonium chloride (62.47 mg,1.1679 mmol). The reaction was carried out at 80℃for 1 hour. After the reaction was completed, the reaction was resumed at room temperature. And (5) filtering. The ethanol was turned off and ethyl acetate (3X 10 mL) was added. And (5) extracting. The organic phase was collected, dried, filtered and concentrated. The concentrate was passed through 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)。
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 (100 mg,0.2814 mmol) was dissolved in tetrahydrofuran (5 mL), and 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester 206e (188.72 mg,0.5628 mmol) and triethylamine (56.95 mg,0.5628 mmol) were added. The reaction was carried out at room temperature for 16 hours. After the reaction was completed. Spin-drying the solvent, and passing the concentrate through a silica gel column (petroleum ether/ethyl acetate=2/1) to give dimethyl 4- (3- (4-butoxy-5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2, 3-dicarboxylate 206f (50 mg, yellow solid). Yield: 29.18%.
MS m/z(ESI)：597.5(M+1)。
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
To a mixed solution of 4- (3- (4-butoxy-5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2, 3-dicarboxylic acid dimethyl ester 206f (50 mg,0.0838 mmol) in tetrahydrofuran/water=1/1 (5 mL) was added lithium hydroxide (4.01 mg,0.1676 mmol). The reaction was carried out at room temperature for 1 hour. After the reaction, the solvent was dried by spin drying. The residue was taken up in 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%.
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
First step
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 (150 mg, 0.458 mmol) was dissolved in acetonitrile (5 mL), followed by addition of 1-bromo-2-methoxyethane (127 mg,0.912 mmol), potassium iodide (76 mg, 0.458 mmol) and potassium carbonate (126 mg,0.912 mmol), stirring was carried out at 80 ℃ for 2 hours, after completion of the reaction, the reaction solution 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 a yield of 56.6%.
MS m/z(ESI)：410.1(M+Na)。
Second step
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 (100 mg,0.258 mmol) was dissolved in ethanol/water=4/1 (5 mL), then iron powder (144 mg, 2.552 mmol) was added, the reaction solution was stirred at 80 ℃ for 1 hour, after the reaction was completed, the reaction solution was spin-dried over silica gel column (petroleum ether/ethyl acetate=5/1) to separate and purify 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)。
Third step
Preparation of dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2, 3-dicarboxylate
5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoro-4- (2-methoxyethoxy) aniline 207c (70 mg,0.193 mmol) was dissolved in tetrahydrofuran (3 mL), followed by the addition of dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (71 mg,0.212 mmol) and triethylamine (58 mg,0.579 mmol), the reaction mixture was stirred at room temperature for 16 hours, the reaction was concentrated over a silica gel column (petroleum ether/ethyl acetate=2/1) and isolated and purified to give dimethyl 4- (3- (5- ((5, 3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 207d (85 mg, yellow solid) in 73.6% yield.
MS m/z(ESI)：599.1(M+1)。
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 separated and purified dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy)) -2-fluoro-4- (2-methoxyethoxy) phenyl) ureido) thiophene-2, 3-dicarboxylic acid 207d (85 mg,0.142 mmol) was dissolved in tetrahydrofuran/water=1/1 (2 mL), followed by addition of lithium hydroxide (10 mg,0.426 mmol), the reaction solution was stirred at room temperature for 1 hour, after completion of the reaction, the reaction solution was concentrated to prepare 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
First step
Preparation of 3- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-methoxy-5-nitropyridine
3-bromo-2-methoxy-5-nitropyridine 208a (500 mg,2.1457 mmol) was dissolved in 1, 4-dioxane (10 mL), and (2, 3-difluoro-6-methoxyphenyl) methanol 208b (448.39 mg, 2.57254 mmol), tris (dibenzylideneacetone) dipalladium (123.38 mg,0.21457 mmol), 2- (dicyclohexylphosphine) -3, 6-dimethoxy-2 '-4' -6 '-tris-I-propyl-11' -biphenyl (172.76 mg,0.321855 mmol), cesium carbonate (1398.22 mg,4.2914 mmol) were added. The reaction was carried out at 90℃for 16 hours under nitrogen protection. After the reaction was completed. 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%.
MS m/z(ESI)：327.2(M+1)
Second step
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 (150 mg,0.4598 mmol) was dissolved in a mixed solution of ethanol/water=1/1 (10 mL), and iron powder (51.36 mg,0.9196 mmol) and ammonium chloride (73.78 mg,1.3794 mmol) were added. After the reaction was completed at 80℃for 1 hour, the temperature was returned to room temperature, ethanol was removed by spinning, ethyl acetate was added for extraction, and the organic phase was collected, dried, filtered and concentrated. 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)
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 (90 mg,0.3038 mmol) was dissolved in tetrahydrofuran (5 mL) and 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester 208e (122.25 mg, 0.36458 mmol) and triethylamine (61.48 mg,0.6076 mmol) were added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried by spin-drying, and the residue was subjected to a silica gel column (petroleum ether/ethyl acetate=1/1) to give dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) ureido) thiophene-2, 3-dicarboxylate 208f (10 mg, yellow liquid). Yield: 5.99%.
MS m/z(ESI)：538.4(M+1)。
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
Dimethyl 4- (3- (5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -6-methoxypyridin-3-yl) ureido) thiophene-2, 3-dicarboxylate 208f (10 mg,0.0186 mmol) was dissolved in a tetrahydrofuran/water=1/1 (3 mL) mixed solution, and lithium hydroxide (0.89 mg,0.0372 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried by spin-drying, and the residue was passed through a 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%.
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
First step
Preparation of (4-chloro-5-methoxypyridin-2-yl) carbamic acid tert-butyl ester
4-chloro-5-methoxypyridin-2-amine 209a (400 mg,2.5223 mmol) was dissolved in dichloromethane (5 mL), di-tert-butyl dicarbonate (1100.98 mg,5.0446 mmol) was added, triethylamine (765.69 mg,7.5669 mmol) was reacted at room temperature for 1 hour, after the reaction was completed, water (10 mL) was added, extracted, dried, and concentrated. The concentrate was then passed through 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%.
MS m/z(ESI)：259.1(M+1)。
Second step
Preparation of (4- ((2, 3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) carbamic acid tert-butyl ester
(4-chloro-5-methoxypyridin-2-yl) carbamic acid tert-butyl ester 209b (270 mg,1.0437 mmol) was dissolved in 1, 4-dioxane (10 mL), and (2, 3-difluoro-6-methoxyphenyl) methanol 209c (218.11 mg,1.25244 mmol), tris (dibenzylideneacetone) dipalladium (95.57 mg,0.10437 mmol), 2- (dicyclohexylphosphine) -3, 6-dimethoxy-2 '-4' -6 '-tris-I-propyl-11' -biphenyl (84.03 mg, 0.156015 mmol), cesium carbonate (680.12 mg,2.0874 mmol) was added. The reaction was carried out at 90℃for 16 hours under nitrogen protection. After the reaction was completed. Water and ethyl acetate were added. Extracting, drying and concentrating. The concentrate was subjected to a 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%.
MS m/z(ESI)：397.1(M+1)。
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 (100 mg,0.2523 mmol) was added to a mixed solution of dichloromethane/trifluoroacetic acid=4/1 (5 mL) at 0 ℃ and then stirred at 0 ℃ for 1 hour. After the reaction was completed, the solvent was dried by spin-drying, and the residue was subjected to 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%.
MS m/z(ESI)：297.0(M+1)。
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 (60 mg,0.2025 mmol) was dissolved in tetrahydrofuran (5 mL), and dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate 209f (135.81 mg,0.405 mmol) and triethylamine (40.98 mg,0.405 mmol) were added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried by spin-drying, and the residue was subjected to a silica gel column (petroleum ether/ethyl acetate=1/1) to give 209g (62 mg, yellow liquid) of dimethyl 5- (3- (4- ((2, 3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) ureido) thiophene-2, 3-dicarboxylate. Yield: 56.99%.
MS m/z(ESI)：538.0(M+1)。
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 (62 mg,0.1154 mmol) of dimethyl 5- (3- (4- ((2, 3-difluoro-6-methoxybenzyl) oxy) -5-methoxypyridin-2-yl) ureido) thiophene-2, 3-dicarboxylate was dissolved in a tetrahydrofuran/water=1/1 (5 mL) mixed solution. Lithium hydroxide (5.53 mg,0.2308 mmol) was added and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, the solvent was dried, and the residue was passed through a 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%.
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
First step
Preparation of 2-methoxy-6- (trifluoromethyl) benzaldehyde
A solution of 2-fluoro-6- (trifluoromethyl) benzaldehyde 210a (900 mg,4.66 mmol) and potassium hydroxide (910 mg,16.31 mmol) in methanol (15 mL) was stirred at 25℃for 1 h. After the reaction was completed, water was added thereto, 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 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
Preparation of (2-methoxy-6- (trifluoromethyl) phenyl) methanol
A solution of 2-methoxy-6- (trifluoromethyl) benzaldehyde 210b (890 mg,4.33 mmol) and sodium borohydride (180 mg,4.77 mmol) in methanol (15 mL) was stirred at 25℃for 1 h. After the reaction was completed, water was added thereto, 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 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), yield: 94.58%.
MS m/z(ESI):189.1(M+1-18)。
Third step
Preparation of 2- (bromomethyl) -1-methoxy-3- (trifluoromethyl) benzene
(2-methoxy-6- (trifluoromethyl) phenyl) methanol 210c (1.7 g,0.01 mol) was dissolved in dichloromethane (20 mL). Phosphorus tribromide (2.73 g,0.01 mol) was added dropwise to the reaction at 0deg.C. The reaction was stirred at 0℃for 2 hours. After the reaction was completed, the mixture was quenched with saturated aqueous sodium bicarbonate solution 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) in yield: 30.05%.
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)..
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 (770 mg,2.85 mmol), 4-fluoro-2-methoxy-5-nitrophenol 210e (693 mg,3.70 mmol), potassium carbonate (788 mg,5.70 mmol) and potassium iodide (236 mg,1.42 mmol) in acetonitrile (10 mL) was stirred at 80℃for 16 h. After the reaction was completed, water was added thereto, 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 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), yield: 57.04%.
MS m/z(ESI):376.9(M+1)。
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 (680 mg,1.80 mmol), iron powder (302 mg,5.42 mmol) and ammonium chloride (290 mg,5.42 mmol) in ethanol: water = 1:1 (20 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) to give 210g of 2-fluoro-4-methoxy-5- (2-methoxy-6- (trifluoromethyl) benzyl) oxy) aniline (490 mg, white solid), yield: 70.47%.
MS m/z(ESI):346.0(M+1)。
Sixth step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2, 3-dicarboxylate
210g (470 mg,1.35 mmol) of 2-fluoro-4-methoxy-5- (2-methoxy-6- (trifluoromethyl) benzyl) oxy) aniline, 211h (540 mg,1.62 mmol) of 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester and triethylamine (274 mg,2.71 mmol) were stirred in a solution of tetrahydrofuran (20 mL) at 25℃for 16 h. After the reaction was completed, water was added to quench, 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 dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-6- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 211i (330 mg, red oil) in yield: 37.25%.
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 (310 mg,0.52 mmol) and lithium hydroxide (37.9 mg,1.58 mmol) were combined in tetrahydrofuran: water = 1:1 (14 mL) was stirred at 25℃for 1 hour. After the completion of 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%.
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
First step
Preparation of (2-methoxy-5- (trifluoromethyl) phenyl) methanol
A solution of 2-methoxy-5- (trifluoromethyl) benzaldehyde 211a (400 mg,1.94 mmol) and sodium borohydride (81.14 mg,2.14 mmol) in methanol (6 mL) was stirred at 25℃for 16 h. After the reaction was completed, water was added thereto, 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 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, transparent oil) as a yield: 84.67%.
MS m/z(ESI):189.1(M+1-18)。
Second step
Preparation of 2- (bromomethyl) -1-methoxy-4- (trifluoromethyl) benzene
(2-methoxy-5- (trifluoromethyl) phenyl) methanol 211b (580 mg,2.79 mmol) was dissolved in dichloromethane (10 mL). PBr was incubated at 0 ℃ 3 (909 mg,3.35 mmol) was added dropwise to the reaction. The reaction was stirred at 0℃for 2 hours. After the reaction was completed, the mixture was quenched with saturated aqueous sodium bicarbonate solution 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-4- (trifluoromethyl) benzene 211c (320 mg, transparent oil) in 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).
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 (100 mg,0.37 mmol), 4-fluoro-2-methoxy-5-nitrophenol 211d (90 mg,0.48 mmol), potassium carbonate (102 mg,0.74 mmol) and potassium iodide (30.74 mg,0.18 mmol) in acetonitrile (5 mL) was stirred at 80℃for 24 hours. After the reaction was completed, water was added thereto, 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 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) as a yield: 77.50%.
MS m/z(ESI):376.0(M+1)。
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 (130 mg,0.34 mmol), iron powder (57 mg,1.03 mmol) and ammonium chloride (55 mg,1.03 mmol) in ethanol: water = 1:1 (10 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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%.
MS m/z(ESI):326.1(M+1-20)。
Fifth step
Preparation of dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2, 3-dicarboxylate
A solution of 2-fluoro-4-methoxy-5- (2-methoxy-5- (trifluoromethyl) benzyl) oxy) aniline 211f (80 mg,0.23 mmol), 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylic acid dimethyl ester 211g (92 mg,0.27 mmol) and triethylamine (46 mg,0.46 mmol) in tetrahydrofuran (5 mL) was stirred at 25℃for 1 hour. After the reaction was completed, water was added thereto, 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 purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) to give dimethyl 4- (3- (2-fluoro-4-methoxy-5- ((2-methoxy-5- (trifluoromethyl) benzyl) oxy) phenyl) ureido) thiophene-2, 3-dicarboxylate 211h (100 mg, clear oil) as a yield: 66.32%.
MS m/z(ESI):587.1(M+1)。
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 (170 mg,0.28 mmol) and lithium hydroxide (20.79 mg,0.86 mmol) 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 it, 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 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
First step
Preparation of 2- ((2- (cyclopentyloxy) -4-fluoro-5-nitrophenoxy) methyl) -3, 4-difluoro-1-methoxybenzene
2- ((2, 3-difluoro-6-methoxybenzyl) oxy) -5-fluoro-4-nitrophenol 212a (300 mg,0.91 mmol), bromocyclopentane (407 mg,2.73 mmol), potassium carbonate (251 mg,1.82 mmol) and potassium iodide (75 mg,0.45 mmol) in acetonitrile (5 mL) were stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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%.
MS m/z(ESI):398.1(M+1)。
Second step
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 (120 mg,0.30 mmol)), iron powder (50 mg,0.90 mmol) and ammonium chloride (48 mg,0.90 mmol) in ethanol: water = 1:1 (12 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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), yield: 89.24%.
MS m/z(ESI):368.0(M+1)。
Third step
Preparation of dimethyl 4- (3- (4- (cyclopentyloxy) -5- (2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2, 3-dicarboxylate
Dimethyl 4- (cyclopentyloxy) -5- ((2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluoroaniline 212c (50 mg,0.13 mmol), 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate 212d (54 mg,0.16 mmol) and triethylamine (27 mg,0.27 mmol) were stirred in tetrahydrofuran (4 mL) for 16 hours at 25 ℃. After the reaction was completed, water was added thereto, 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 purified by silica gel column chromatography (mobile phase: methanol/dichloromethane=10/1) to give dimethyl 4- (3- (4- (cyclopentyloxy) -5- (2, 3-difluoro-6-methoxybenzyl) oxy) -2-fluorophenyl) ureido) thiophene-2, 3-dicarboxylate 212e (47 mg, brown oil) in yield: 51.07%.
MS m/z(ESI):609.2(M+1)。
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 (37 mg,0.06 mmol) and lithium hydroxide (4 mg,0.18 mmol) were combined 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%.
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
First step
Preparation of 2, 3-dihydro-1, 4-benzodioxane-5-methyl alcohol
To a solution of 2, 3-dihydrobenzo [ b ] [1,4] dioxine-5-carboxylic acid 213a (1 g,0.01 mol) in tetrahydrofuran (12 mL) was added dropwise lithium aluminum hydride (15 mL) at 0deg.C. The reaction mixture was stirred at 0℃for 10 minutes and at 25℃for a further 16 hours. After the reaction was completed, water was added thereto, 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 purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1) to give 2, 3-dihydro-1, 4-benzodioxane-5-methyl alcohol 213b (970 mg, white solid), yield: 94.64%.
MS m/z(ESI):149.2(M+1-18)。
Second step
Preparation of 5-bromomethyl-2, 3-dihydro-1, 4-benzodioxine
Phosphorus tribromide (1.7 g,6.28 mmol) was added dropwise to a solution of 2, 3-dihydro-1, 4-benzodioxane-5-methyl alcohol 213b (870 mg,5.23 mmol) in dichloromethane (10 mL) and stirred at 0deg.C for 1 hr. After the reaction was completed, the mixture was quenched with 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), 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).
Third step
Preparation of 5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxin
A solution of 5-bromomethyl-2, 3-dihydro-1, 4-benzodioxine 213c (900 mg,3.92 mol), 4-fluoro-2-methoxy-5-nitrophenol 213d (955 mg,5.10 mmol), potassium carbonate (1.08 g,7.85 mmol) and potassium iodide (326 mg,1.96 mmol) in acetonitrile (10 mL) was stirred at 80℃for 16 hours. After the reaction was completed, water was added thereto, 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 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%.
MS m/z(ESI):(M+1+23)。
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 (720 mg,2.41 mmol), iron powder (404 mg,7.24 mmol) and ammonium chloride (387 mg,7.24 mmol) in ethanol: water = 1:1 (12 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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%.
MS m/z(ESI):306.1(M+1)。
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
Dimethyl 5- ((2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyaniline 213f (750 mg,1.86 mmol), 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate 213g (751mg, 2.24 mmol) and triethylamine (377 mg,3.73 mmol) were stirred in tetrahydrofuran (15 mL) for 16 hours at 25 ℃. After the reaction was completed, water was added to quench, 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 dimethyl 4- (3- (5- ((2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 213h (600 mg, brown oil), yield: 51.42%.
MS m/z(ESI):547.1(M+1)。
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
Dimethyl 4- (3- (5- ((2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 213h (500 mg,0.91 mmol), and lithium hydroxide (65 mg,2.74 mmol) in tetrahydrofuran: water = 1:1 (14 mL) was stirred at 25℃for 1 hour. After the completion of the reaction, the reaction mixture 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%.
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
First step
Preparation of 2- (2-bromo-4-fluorophenoxy) ethan-1-ol
2-bromo-4-fluorophenol 1 (5 g,0.0262 mol), 2-bromoethanol 2 (3.93 g,0.0314 mol) was dissolved in acetonitrile (50 mL), followed by addition of potassium carbonate (7.24 g,0.0524 mol) and potassium iodide (0.43 g,0.0026 mol), stirring of the reaction solution at 80 ℃ for 16 hours, after completion of the reaction, suction filtration of the reaction solution, concentration of the filtrate, and purification by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=10/1) to give 2- (2-bromo-4-fluorophenoxy) ethane-1-ol 3 (5 g, white solid), yield: 77.10%.
MS m/z(ESI):236.1(M+1)。
Second step
Preparation of 6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin
2- (2-bromo-4-fluorophenoxy) ethan-1-ol 3 (1.2 g,0.0051 mmol), palladium acetate (0.02 mg,0.0001 mmol), t BuDavePhos (0.04 g,0.00012 mmol), cesium carbonate (2.49 g,0.0077 mmol) was added to toluene (20 mL), the reaction was carried out at 80℃for 1 hour by microwave, after the reaction was completed, the reaction solution 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 ] ][1,4]Dioxin 4 (0.5 g, white solid), yield: 62.75%.
MS m/z(ESI):155.1(M+1)。
Third step
Preparation of 6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-carbaldehyde
6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin 4 (200 mg,1.2975 mmol) was dissolved in tetrahydrofuran (10 mL), 2M LDA tetrahydrofuran solution (5.2 mL) was slowly dropped at-78℃and reacted for 30 minutes, DMF (189.67 mg,2.595 mmol) was slowly dropped, stirring was continued for 30 minutes, after the reaction was completed, an aqueous ammonium chloride solution was added to the reaction solution, ethyl acetate was extracted, and 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-carbaldehyde 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).
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 (200 mg,1.098 mmol) was dissolved in methanol (10 mL), sodium borohydride (29.08 mg,0.768 mmol) was slowly added at 0 ℃ after 1 hour of reaction, most of the methanol was removed by spinning, aqueous ammonium chloride solution was added, dichloromethane extraction was performed, and 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), yield: 89.02%.
MS m/z(ESI):185.2(M+1)。
Fifth step
Preparation of 5- (bromomethyl) -6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin
(6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (180 mg,0.9774 mmol) was dissolved in methylene chloride (10 mL), phosphine tribromide (264.88 mg,0.9774 mmol) was added at 0℃and the reaction mixture was stirred at room temperature for 1 hour, after the completion of the reaction, the reaction mixture 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), yield: 22.72%.
MS m/z(ESI):247.1(M+1)。
Sixth step
Preparation of 6-fluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxin
A solution of 5- (bromomethyl) -6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxine 7 (52 mg,0.2105 mmol), 4-fluoro-2-methoxy-5-nitrophenol 8 (47.27 mg,0.2526 mmol), potassium carbonate (58.19 mg, 0.426 mmol) and potassium iodide (3.49 mg,0.021 mmol) in acetonitrile (5 mL) was stirred at 80℃for 16 hours. After the reaction was completed, water was added thereto, 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 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), yield: 43.04%.
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 (30 mg,0.0849 mmol), iron powder (14.22 mg,0.2547 mmol) and ammonium chloride (9.08 mg,0.1698 mmol) in ethanol: water = 1:1 (6 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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) in yield: 72.91%.
MS m/z(ESI):324.3(M+1)。
Eighth step
Preparation of dimethyl 4- (3- (5- ((6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
Dimethyl 2-fluoro-5- ((6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -4-methoxyaniline 10 (20 mg,0.0619 mmol), 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate 11 (31.14 mg,0.0929 mmol) and triethylamine (12.53 mg,0.1238 mmol) in tetrahydrofuran (5 mL) were stirred at 25 ℃ for 16 hours. After the reaction was completed, water was added to quench, 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 dimethyl 4- (3- (5- ((6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 12 (16 mg, yellow solid), yield: 45.72%.
MS m/z(ESI)：565.5(M+1)。
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
Dimethyl 4- (3- (5- ((6-fluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 12 (16 mg,0.0283 mmol) and lithium hydroxide (2.03 mg,0.0849 mmol) in tetrahydrofuran: water = 1:1 (8 mL) was stirred at 25℃for 1 hour. After the completion of the reaction, water was added thereto, the mixture was quenched, extracted with ethyl acetate (3×10 mL), and 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%.
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
First step
Preparation of 2- (2-bromo-4, 5-difluorophenoxy) ethyl-1-ol
2-bromo-4, 5-difluorophenol 1 (5 g,0.0239 mol), 2-bromoethanol (5.97 g,0.0478 mol) was dissolved in acetonitrile (50 mL), followed by addition of potassium carbonate (6.61 g,0.0478 mol) and potassium iodide (3.97 g,0.0239 mol), 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 concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=10/1) to give 2- (2-bromo-4, 5-difluorophenoxy) ethyl-1-ol 3 (5 g, white solid), yield: 82.68%.
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
Preparation of 6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin
To 2- (2-bromo-4, 5-difluorophenoxy) ethyl-1-ol 3 (4 g,15.8 mmol), palladium acetate (355 mg,1.58 mmol), t BuDavePhos (1.08 g,3.16 mmol), cesium carbonate (10.3 g,31.6 mmol) was added to toluene (20 mL), the reaction was carried out at 80℃for 1 hour by microwave, after the reaction was completed, the reaction solution 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, 7-difluoro-2, 3-dihydrobenzo [ b ] after purification][1,4]Dioxin 4 (1 g, white solid), yield: 36.79%.
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.
Third step
Preparation of 6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-carbaldehyde
Dissolving 6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin 4 (1 g,5.81 mmol) in tetrahydrofuran (10 mL), slowly dropwise adding 2M LDA tetrahydrofuran solution (5.8 mL) at minus 78 ℃ for 30 minutes, slowly dropwise adding DMF (0.51 g,6.96 mmol) after reaction, continuously stirring for 30 minutes, adding ammonium chloride aqueous solution to the reaction solution after reaction is finished, extracting by ethyl acetate, concentrating an organic phase, purifying by silica gel column chromatography (mobile phase: petroleum ether/ethyl acetate=5/1), and obtaining 6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-formaldehyde 5 (1 g, white solid), yield: 86%.
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).
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,4.7 mmol) was dissolved in methanol (10 mL), sodium borohydride (378 mg,9.99 mmol) was slowly added at 0 ℃ and after 1 hour of reaction, most of the methanol was removed by spinning, aqueous ammonium chloride solution was added, dichloromethane extraction was performed, and 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), yield: 94.72%.
MS m/z(ESI):185.0(M-17)。
Fifth step
Preparation of 5- (bromomethyl) -6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin
(6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin-5-yl) methanol 6 (900 mg,4.45 mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (1.4 g,5.34 mmol) was added, carbon tetrabromide (1.77 g,5.34 mmol) was added after stirring for a while, 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, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin 7 (800 mg, white solid), 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).
Sixth step
Preparation of 6, 7-difluoro-5- ((4-fluoro-2-methoxy-5-nitrophenoxy) methyl) -2, 3-dihydrobenzo [ b ] [1,4] dioxin
A solution of 5- (bromomethyl) -6, 7-difluoro-2, 3-dihydrobenzo [ b ] [1,4] dioxin 7 (300 mg,1.13 mmol), 4-fluoro-2-methoxy-5-nitrophenol 8 (211 mg,1.13 mmol), potassium carbonate (312 mg,2.26 mmol) and potassium iodide (93 mg,0.56 mmol) in acetonitrile (5 mL) was stirred at 80℃for 16 hours. After the reaction was completed, water was added thereto, 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 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 (340 mg,0.91 mmol), iron powder (153 mg,2.74 mmol) and ammonium chloride (146 mg,2.74 mmol) in ethanol: water = 1:1 (6 mL) was stirred at 80℃for 1 hour. After the reaction was completed, water was added thereto, 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 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 (250 mg,0.73 mmol), dimethyl 4- (phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate 11 (254 mg,0.87 mmol) and triethylamine (148 mg,1.46 mmol) in tetrahydrofuran (5 mL) were stirred at 25 ℃ for 16 hours. After the reaction was completed, water was added to quench, 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 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 12 (580 mg, yellow solid), yield: 95.15%.
MS m/z(ESI)：583.0(M+1)。
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 (520 mg,0.89 mmol) and lithium hydroxide (64 mg,2.67 mmol) in tetrahydrofuran: water = 1:1 (8 mL) was stirred at 25℃for 1 hour. After the completion of the reaction, water was added thereto, the mixture was quenched, extracted with ethyl acetate (3×10 mL), and 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%.
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
First step
Preparation of 1-bromo-4-chloro-2- (2, 2-diethoxyethoxy) benzene
2-bromo-5-chlorophenol 216a (10 g,48.2 mmol) was dissolved in DMF (100 mL), followed by addition of 2-bromo-1, 1-diethoxyethane (9.5 g,48.2 mmol), potassium carbonate (13.3 g,96.4 mmol), stirring at 100 ℃ for 16 hours, after completion of the reaction, the reaction solution was filtered, washed with water, extracted with ethyl acetate, and the organic phase was dried and purified by silica gel column (petroleum ether/ethyl acetate=10/1) to give 1-bromo-4-chloro-2- (2, 2-diethoxyethoxy) benzene 216b (11 g, white solid) in 70.5% yield.
MS m/z(ESI)：347.1(M+Na)。
Second step
Preparation of 7-bromo-4-chlorobenzofuran
1-bromo-4-chloro-2- (2, 2-diethoxyethoxy) benzene 216b (11 g,34.0 mmol) was dissolved in toluene (100 mL), PPA (11.5 g,34.0 mmol) was then added, stirring was carried out at 80℃for 2 hours, after the reaction was completed, ice water was added to the reaction solution, extraction was carried out with ethyl acetate, and the organic phase was dried and spin-dried and purified by silica gel column (petroleum ether) to give 7-bromo-4-chlorobenzofuran 216c (5.7 g, white solid) in 72.4% yield.
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).
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 (520 mg,4.92 mmol) was added thereto, the reaction mixture was stirred under hydrogen pressure at 25℃for 16 hours, after the completion of the reaction, the reaction mixture was filtered, and the filtrate was dried by spin-drying over a silica gel column (petroleum ether) to obtain 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).
Fourth step
Preparation of methyl 4-chloro-2, 3-dihydrobenzofuran-7-carboxylate
7-bromo-4-chloro-2, 3-dihydrobenzofuran 216d (2 g,8.6 mmol) was dissolved in methanol (100 mL), pd (dppf) Cl2 (0.63 g,0.86 mmol) and potassium acetate (1.69 g,17.2 mmol) were then added and stirred in a carbon monoxide atmosphere at 80℃for 16 hours, after the reaction was completed, the reaction solution was filtered, and the filtrate was dried by spin-drying over a silica gel column (petroleum ether/ethyl acetate=10/1) to give methyl 4-chloro-2, 3-dihydrobenzofuran-7-carboxylate 216e (650 mg, white solid) in 35.5% yield.
MS m/z(ESI)：213.1(M+H)。
Fifth step
Preparation of (4-chloro-2, 3-dihydrobenzofuran-7-yl) methanol
Methyl 4-chloro-2, 3-dihydrobenzofuran-7-carboxylate 216e (650 mg,3.1 mmol) was dissolved in tetrahydrofuran (10 mL), lithium aluminum hydrogen (235 mg,6.2 mmol) was added at 0 ℃ and stirred for 1 hour, after the reaction was completed, an aqueous ammonium chloride solution was added to the reaction solution, ethyl acetate was extracted, and the organic phase was dried and spin-dried and purified by silica gel column (petroleum ether/ethyl acetate=5/1) to give (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)。
Sixth step
Preparation of 7- (bromomethyl) -4-chloro-2, 3-dihydrobenzofuran
(4-chloro-2, 3-dihydrobenzofuran-7-yl) methanol 216f (400 mg,2.2 mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (682 mg,2.6 mmol) was added at 0deg.C, and after stirring for 10 min, carbon tetrabromide (862 mg,2.6 mmol) was added and the reaction stirred at room temperature for 1 hour. After the reaction, the reaction mixture was dried by spin-drying and separated and purified by a silica gel column (petroleum ether) to give 216g (380 mg, white solid) of 7- (bromomethyl) -4-chloro-2, 3-dihydrobenzofuran as a preparation with a yield of 69.8%.
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 (380 mg,1.5 mmol) and 4-fluoro-2-methoxy-5-nitrophenol (287 mg,1.5 mmol) were dissolved in acetonitrile (10 mL), followed by addition of potassium carbonate (284 mg,3.1 mmol), potassium iodide (255 mg,1.5 mmol), stirring of the reaction solution at 70 ℃ for 16 hours, after completion of the reaction, addition of water to the reaction solution, extraction with ethyl acetate, drying of the organic phase, concentration and separation and purification 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) in 22.6% yield.
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 (120 mg,0.34 mmol) was dissolved in ethanol/aqueous ammonium chloride=4/1 (10 mL), iron powder (189 mg,3.4 mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after the reaction was completed, the reaction solution was filtered, the filtrate was concentrated and separated and purified by a silica gel column (petroleum ether/ethyl acetate=3/1) to give 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)。
Ninth step
Preparation of dimethyl 4- (3- (5- ((4-chloro-2, 3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate
5- ((4-chloro-2, 3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyaniline 216i (100 mg,0.31 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (114 mg,0.34 mmol) and triethylamine (94 mg,0.93 mmol), the reaction mixture was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction mixture was concentrated and purified by column on silica gel (petroleum ether/ethyl acetate=3/1) to give dimethyl 4- (3- (5- ((4-chloro-2, 3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 216j (150 mg, yellow solid) in 85.6% yield.
MS m/z(ESI)：565.0(M+H)。
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
Dimethyl 4- (3- (5- ((4-chloro-2, 3-dihydrobenzofuran-7-yl) methoxy) -2-fluoro-4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 216j (150 mg,0.26 mmol) was dissolved in tetrahydrofuran/water=1/1 (10 mL), lithium hydroxide (19 mg,0.78 mmol) was added, the reaction mixture was stirred at room temperature for 1 hour, and after completion of the reaction, the reaction was dried, and 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) was prepared 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
First step
Preparation of 2-bromo-1- (2, 2-diethoxyethoxy) -3-fluorobenzene
A solution of 2-bromo-3-fluorophenol 217a (10 g,0.05 mol) in DMF (30 mL) was added dropwise to sodium hydride and the reaction mixture stirred at 25℃for 1 hour. 2-bromo-1, 1-dioxyethane (11.36 g,0.05 mol) was then added to the reaction mixture, stirred at 153 ℃ for 2 hours, after the reaction was completed, the reaction solution was suction-filtered, the filtrate was concentrated, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=19/1) to give 2-bromo-1- (2, 2-diethoxyethoxy) -3-fluorobenzene 217b (9.4 g, colorless liquid), yield: 52.5%.
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
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 (1.55 g,4.6 mmol) was added chlorobenzene (20 mL) and the reaction mixture was stirred at 80℃for 2 hours. After the reaction was completed, water was added thereto, 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 purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/1) to give 7-bromo-6-fluorobenzofuran 217c (680 mg, white solid), yield: 60.9%.
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).
Third step
Preparation of 7-bromo-6-fluoro-2, 3-dihydrobenzofuran
7-bromo-6-fluorobenzofuran 217C (2.6 g,12.1 mmol) was dissolved in ethanol (50 mL), rh/C (620 mg,6.1 mmol) was then added, the reaction mixture was stirred under hydrogen pressure at 25℃for 16 hours, after the reaction was completed, the reaction mixture was filtered, and the filtrate was spin-dried 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).
Fourth step
Preparation of 6-fluoro-2, 3-dihydrobenzofuran-7-carbaldehyde
7-bromo-6-fluoro-2, 3-dihydrobenzofuran 217d (1.25 g,5.8 mmol) was dissolved in tetrahydrofuran (20 mL), isopropyl magnesium chloride (420 mg,2.9 mmol) was added at 0deg.C, stirred for 30 min, then n-butyllithium (560 mg,8.7 mmol) was added, stirred for 30 min, finally DMF (470 mg,6.4 mmol) was added, stirred for 30 min, after the reaction was completed, saturated ammonium chloride was added, ethyl acetate was extracted, and the organic phase was spin-dried over silica gel column (petroleum ether/ethyl acetate=5/1) 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)。
Fifth step
Preparation of (6-fluoro-2, 3-dihydrobenzofuran-7-yl) methanol
6-fluoro-2, 3-dihydrobenzofuran-7-carbaldehyde 217e (700 mg,4.2 mmol) was dissolved in tetrahydrofuran (10 mL), lithium aluminum hydrogen (320 mg,8.2 mmol) was added at 0deg.C and stirred for 1 hour, after the completion of the reaction, an aqueous ammonium chloride solution was added to the reaction solution, ethyl acetate was used for extraction, and the organic phase was dried and spun-dried over a silica gel column (petroleum ether/ethyl acetate=3/1) to obtain (6-fluoro-2, 3-dihydrobenzofuran-7-yl) methanol 217f (400 mg, white solid) in a yield of 56.6%.
MS m/z(ESI)：151.1(M-17)。
Sixth step
Preparation of 7- (bromomethyl) -6-fluoro-2, 3-dihydrobenzofuran
(6-fluoro-2, 3-dihydrobenzofuran-7-yl) methanol 217f (400 mg,2.4 mmol) was dissolved in dichloromethane (10 mL), triphenylphosphine (749 mg,2.8 mmol) was added at 0deg.C, and after stirring for 10 min, carbon tetrabromide (946 mg,2.8 mmol) was added, and the reaction was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was dried by spin-drying and separated and purified by a silica gel column (petroleum ether) to give 217g (250 mg, white solid) of 7- (bromomethyl) -6-fluoro-2, 3-dihydrobenzofuran as a preparation in 45.1% yield.
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 (206 mg,1.1 mmol) was dissolved in DMF (3 mL), then sodium hydrogen (32 mg,1.3 mmol) was added at 0deg.C, the reaction mixture was stirred for 1 hour, then 217g (250 mg,1.1 mmol) of 7- (bromomethyl) -6-fluoro-2, 3-dihydrobenzofuran was prepared, the reaction mixture was stirred for 1 hour at 0deg.C, water was added to the reaction mixture after the completion of the reaction, ethyl acetate was extracted, the organic phase was concentrated by dryness and then separated and purified by 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 40.4% yield.
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 (150 mg,0.44 mmol) was dissolved in ethanol/aqueous ammonium chloride=4/1 (10 mL), iron powder (246 mg,4.4 mmol) was added, the reaction was reacted at 80 ℃ for 2 hours, after the reaction was completed, the reaction solution was filtered, the filtrate was concentrated and after separation and purification by silica gel column (petroleum ether/ethyl acetate=3/1) was carried out 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)。
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 (120 mg,0.39 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the addition of dimethyl 4- ((phenoxycarbonyl) amino) thiophene-2, 3-dicarboxylate (144 mg,0.43 mmol) and triethylamine (118 mg,1.17 mmol), the reaction mixture was stirred at 40 ℃ for 16 hours, after completion of the reaction, the reaction mixture was concentrated and purified by column on silica gel (petroleum ether/ethyl acetate=3/1) to give dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-2, 3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylate 217j (180 mg, yellow solid) in 84.2% yield.
MS m/z(ESI)：549.1(M+H)。
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
Dimethyl 4- (3- (2-fluoro-5- ((6-fluoro-2, 3-dihydrobenzofuran-7-yl) methoxy) -4-methoxyphenyl) ureido) thiophene-2, 3-dicarboxylic acid 217j (180 mg,0.33 mmol) was dissolved in tetrahydrofuran/water=1/1 (10 mL), lithium hydroxide (41 mg,0.99 mmol) was added, the reaction mixture was stirred at room temperature for 1 hour, and after completion of the reaction, the reaction was dried and 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) was prepared in 50.7% yield.
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 determination of the Activity of the inventive Compounds on human GnRHR
The method is used for determining antagonism of the compounds of the invention on the activity of the human GnRHR protein expressed in the human GnRHR/HEK293 stably transformed cell line.
1. Test material and instrument
1.1 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 instrument consumables
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 human GnRHR/HEK293 stably transfected cell lines were inoculated onto 384-well cell culture plates at a density of 12000 cells/well/25. Mu.l, at 37℃with 5% CO 2 Culturing overnight;
2.2 freeze thawing 20 XComponent A to room temperature, diluting it to 2 Xworking concentration with Assay Buffer, and standing at room temperature;
2.3 cell culture plates were equilibrated at room temperature for 10min, medium removed, added to 20. Mu.L Assay Buffer and 20. Mu.L 2X Component A,200g,RT, centrifuged for 3-5sec, and incubated at 37℃for 2 h;
2.4 compounds were 3-fold diluted in 3834 pp_dmso plates using DMSO, then transferred 240nl/well each to the working plate using Echo 550, 200g, rt,1min; adding 40 μl of Assay Buffer into a working plate, 200g, RT,1min, mixing with vibrator 2500rpm,20min, 200g, RT,1min for use;
2.5 preparing 180nM LH-RH (6X) with an Assay Buffer, and placing in 50ul to 3657 plates for use;
2.6 taking out the cell culture plate, standing at room temperature for 10min, adding 10 μl of the diluted compound in step 2.4 into the corresponding well, and standing at 25deg.C for 30min;
2.7 10. Mu.l of the diluted compound from step 2.5 were added to the corresponding experimental wells using FLIPR Tetra and the data collected.
3. Experimental data:
the antagonistic activity of the compound of the present invention on human GnRHR is determined by the above experiment, and the IC is determined 50 The values are shown in Table 1.
TABLE 1 IC of the antagonistic Activity of the compounds of the invention against human GnRHR 50 Value of
/>
4. Conclusion of experiment:
the above data show that the compounds of the present invention have significant antagonism on human GnRHR activity.
Test example 2 hERG inhibition test of the Compounds of the invention
The rapid activation of the human ventricle delays the rectified potassium current (IKr) is mediated primarily by the human ether-a-go-go related gene (hERG) potassium ion channel in the heart. Inhibition of IKr is the most common mechanism of ventricular muscle action potential prolongation caused by non-heart-like drugs. Prolongation of the action potential time course will lead to prolongation of QT interval on clinical electrocardiograms, which is associated with dangerous ventricular arrhythmias and torsades de pointes. Because of these cardiotoxic effects, many drugs have been withdrawn from later clinical trials, it is important to identify inhibitors early in drug development. Thus, studying the effect of drugs on hERG potassium channels in a system of heterologous expression is a non-clinical study recommended by ICH as a test drug for the effect on QT interval.
hERG inhibition studies aim 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 with human ether-a-go-related gene (hERG).
The cell seed was placed on the inverted microscope stage in a cell recording tank, and one cell in the recording tank was randomly selected for the test. The perfusion system was mounted on an inverted microscope stage and used to continuously perfuse cells with ECS.
Manual patch clamp test recording microelectrodes were prepared with capillary glass tubes, in which intracellular fluid was filled. On the day of patch clamp testing, electrodes were prepared using borosilicate glass tubing (GC 150TF-10,Harvard Apparatus Co.UK). The resistance after filling the electrodes with ICS is between 2-5MΩ.
The clamp voltage was-80 mV, the first depolarization to +60mV and maintenance of 850ms open hERG channel. The voltage is then set to-50 mV and maintained 1275ms, producing a bounce current or tail current, the peak of which will be measured and used for analysis. Finally, the voltage is restored to the clamp voltage (-80 mV). This command voltage sequence was repeated every 15s during the test.
And in the beginning stage of the record of the perfusion of the solvent control working solution, monitoring the tail current peak value until more than 3 scanning curves are stabilized, and then, the compound working solution to be tested can be perfused until the inhibition effect of the compound working solution on the hERG current peak value reaches a stable state. The most recent continuous 3 current curve peaks are basically coincident to be used as a standard for judging whether the state is stable, and the next concentration of compound is continuously poured after the stable state is reached.
For each concentration all percent inhibition recorded was averaged, IC 50 The value is determined by the concentration effect curveIs obtained by the method. IC (integrated circuit) 50 All compounds with values above 10 μm are not considered potent inhibitors of hERG channel, but have ICs below 1 μm 50 Compounds of this value are then considered potent hERG channel inhibitors.
Experimental data:
TABLE 2 IC of the compounds of the invention in hERG inhibition assays 50 Values.
Conclusion of experiment:
none of the above data shows that the compounds of the present invention are potent inhibitors of the hERG channel.
Test example 3 rat pharmacokinetic test of the Compounds of the invention
1. The purpose of the experiment is as follows:
the concentration of the drug in plasma at various times after the compound of the examples was administered by gavage in rats, which were determined by LC/MS using SD rats as the test animals. The pharmacokinetic behavior of the compound of the invention in rats was studied and its pharmacokinetic profile was evaluated.
2. Experimental protocol
2.1 Experimental drugs
The compound of the embodiment of the invention is self-made.
2.2 laboratory animals
Healthy adult SD rats.
2.3 pharmaceutical formulation
An appropriate amount of sample is weighed, and the formulation of the gastric administration group solvent is 2% DMA+97.4% (0.5% CMC-Na) +0.6%2M NaOH, and the solution is prepared into a suspension of 0.3mg/mL by ultrasonic treatment.
2.4 administration of drugs
SD rats were administered by gastric lavage after overnight fast at a dose of 3mg/kg.
3. The experimental steps are as follows:
the stomach-irrigating administration group is subjected to blood sampling for 0.1mL before administration and 30min after administration, 1h, 4h, 6h, 8h, 12h, 24h, 48h, 72h, 96h, 168h and 240h, placed in a heparinized test tube, centrifuged at 3500rpm for 10 min, and the plasma is separated and stored at-20 ℃. Feeding/free drinking water was resumed 4 hours after dosing.
The LC/MS method was used to determine the amount of test compound in the plasma of rats following gavage administration.
4. Experimental data:
table 3 pharmacokinetic parameters of the compounds of the present invention.
5. Conclusion of experiment:
the data show that the compound of the invention has better absorption of the rat gastric lavage drug substitution, obviously prolonged half-life and obvious drug substitution 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, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (12)

1. A compound of formula I-3 or a pharmaceutically acceptable salt thereof:
wherein,
e is selected from S (O) 2 、OCH 2 Or NHCH 2 ；
G is selected from unsubstituted or optionally substituted by one, two or more R 4 Substituted 9-10 membered heteroaryl;
j is selected from H, C 1-6 An alkyl group;
R 1 selected from COOH;
each R 4 The same or different, independently of one another, from hydrogen, halogen, C 1-3 Alkyl, C 1-3 An alkyl oxy group.
2. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, having the structure according to formula II:
wherein,
E. g has the definition of claim 1.
3. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, having the structure according to formula III:
wherein,
g has the definition of claim 1.
4. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein G is selected from the group consisting of:
5. a compound as shown below or a pharmaceutically acceptable salt thereof:
6. a process for the preparation of a compound of formula I-3 as claimed in any one of claims 1 to 5, comprising scheme 1 or scheme 2 below: scheme 1: the compound 1 undergoes cyclization under the action of alkali to obtain a compound of a formula I-3;
therein, E, G, J, R 1 Having the definition as claimed in any one of claims 1 to 5, R is C 1-3 An alkyl group;
and/or, the base in scheme 1 is selected from lithium hydroxide, lithium hydroxide monohydrate, sodium hydroxide, potassium hydroxide;
Scheme 2: reacting the compound 2 with a compound 3 to obtain a compound shown in a formula I-3;
therein, E, G, J, R 1 Having the definition of any one of claims 1 to 5;
y enables compound 3 to be substituted or coupled to compound 2 in a reaction selected from halogen, NH 2 、B(OH) 2 ；
And/or, the reaction in scheme 2 is carried out in the presence of a base selected from at least one of triethylamine, diisopropylethylamine, pyridine, DMAP, DBU; or at least one of lithium diisopropylamide, methyl lithium and butyl lithium;
and/or the reaction of scheme 1 or scheme 2 is carried out in the presence of an organic solvent or a mixed solvent of an organic solvent and water.
7. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of any one of claims 1-5 or a pharmaceutically acceptable salt thereof.
8. Use of a compound according to any one of claims 1 to 5 or at least one of its pharmaceutically acceptable salts, and a pharmaceutical composition according to claim 7 for the manufacture of a medicament for the treatment of a gonadotropin-related disorder.
9. Use of a compound according to any one of claims 1 to 5, or at least one of its pharmaceutically acceptable salts, and a pharmaceutical composition according to claim 7 for the manufacture of a medicament for a GnRH receptor antagonist.
10. Use of at least one of the compounds of any one of claims 1-5 or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition of claim 7 for 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 fibroids, endometriosis, uterine fibroids, precocious puberty, amenorrhea, premenstrual syndrome, dysmenorrhea, polycystic ovary syndrome, acne, alopecia, alzheimer's disease, infertility, irritable bowel syndrome, diseases that are hormone independent and LH-RH sensitive benign or malignant tumors or flushing.
11. Use of a compound according to any one of claims 1 to 5 or at least one of its pharmaceutically acceptable salts, and a pharmaceutical composition according to claim 7 for the preparation of a reproductive modulator, a contraceptive, an ovulation inducer; or as a medicament for the prevention of postoperative recurrence of sex hormone dependent cancer.
12. The use according to claim 11, wherein the sex hormone dependent cancer is selected from prostate cancer, uterine tumor, breast cancer, pituitary cancer.
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