HK1196820B - Novel heterocyclic derivatives and their uses - Google Patents
Novel heterocyclic derivatives and their uses Download PDFInfo
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- HK1196820B HK1196820B HK14110286.6A HK14110286A HK1196820B HK 1196820 B HK1196820 B HK 1196820B HK 14110286 A HK14110286 A HK 14110286A HK 1196820 B HK1196820 B HK 1196820B
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Description
Technical Field
The present invention relates to novel heterocyclic compounds for the preparation of medicaments for the treatment of diseases which are associated with different functions of the histamine 4 receptor. In particular, the medicament is for the treatment of inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye disease, cardiac dysfunction, cardiac arrhythmias, atherosclerosis, multiple sclerosis, inflammatory bowel diseases (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, post-operative adhesions, vestibular disorders and cancer.
Background
Histamine is an biogenic amine which plays a central role in immune and inflammatory responses and is also a neurotransmitter. For example, histamine controls different functions of cytokine secretion in antigen presenting cells (dendritic cells and macrophages), T cells, B cells, epithelial and endothelial cells, T cell proliferation or dendritic cells and mast cells (JDDG, 2010, 8, 495-. There are 4 histamine receptors (histamine 1 receptor, histamine 2 receptor, histamine 3 receptor, and histamine 4 receptor) (br.j. pharm2006, 147, S127-S135). Acute allergic reactions are controlled by histamine 1 receptors distributed throughout the body (br.j.pharmac.chemither.1966, 27, 427- ° 439), while gastric acid secretion is controlled by histamine 2 receptors also distributed throughout the body like histamine 1 receptors (Nature1972, 236, 385-. It is well known that neurotransmitter secretion in the central nervous system is controlled by histamine 3 receptors expressed in neurons (Nature1983, 302, 832-Asn 837). The histamine 4 receptor further explains many physiological functions of signaling processes that cannot be explained with the histamine 1 receptor, the histamine 2 receptor and the histamine 3 receptor alone. The histamine 4 receptor was first reported in 1994 and its cloning has only been performed since 2000. The histamine 4 receptor is a G-protein coupled receptor consisting of 390 amino acids and activated to increase calcium concentration or inhibit cyclic adenosine monophosphate (cAMP) by binding to Gi/o protein (The Open Immunology Journal, 2009, 2, 9-41). The histamine 4 receptor is mainly expressed in bone marrow or eosinophils, basophils, T cells, mast cells, monocytes and dendritic cells, and is also observed in spleen, thymus, lung, heart and intestine (nat. rev. drug discov.2008, 7, 41-53; biohem. biophy. res. commu.2000, 279, 615-. The histamine 4 receptor not only plays a central role in immune responses, but also has effects on the activation and migration of different immune cells, the production of cytokines and chemokines (J.Immunol.2005, 174, 5224-.
In different in vivo experiments, the histamine 4 receptor is known to play an important role in inflammation and itch (J.allergy Clin.Immunol.2007, 119, 176-. Particularly as a result of the study, it has been found that a histamine 4 antagonist relieves pneumonia by controlling Th2(T helper type 2) reaction in an allergic mouse asthma model, and it is confirmed that the histamine 4 antagonist effectively suppresses histamine-induced itch. This dual effect on allergic inflammation and itching is the basis of the fact that the histamine 4 receptor can be a good target for the treatment of allergic skin diseases such as atopic dermatitis (j. invest. dermatol.2010, 130(4), 1023- & 1033).
Antagonism of the diverse functions of histamine 4 receptors in such immune cells is a key focus for the study of inflammatory diseases, pruritus, pain, allergic rhinitis, asthma, rheumatoid arthritis, atopic dermatitis, idiopathic chronic urticaria, inflammatory pain, neuropathic pain and osteoarthritic pain. Furthermore, recent studies involving the effectiveness of the histamine 4 receptor on cancer have been announced, and thus it is expected to be developed as an anticancer drug.
Recently, quinoxaline-based derivatives have been reported in WO2010/030785 to exhibit activity on the histamine 4 receptor. However, they do not show sufficient pharmacological activity in vivo in animal models because of their poor solubility and metabolic stability.
Disclosure of the invention
Technical problem
The heterocyclic compounds of the present invention (including pyridopyrazines, pyridopyrimidines, and naphthyridines) exhibit histamine 4 receptor inhibitory activity that is the same as or greater than that of conventional human histamine 4 receptor (hH4R) inhibitors such as those disclosed in WO 2010/030785; exhibit selectivity for histamine receptors of each subtype and receptors, transporters and ion channels on membranes; have higher solubility, metabolic stability and thus potent pharmacokinetic properties, and are therefore useful for treatment at lower doses and with fewer dosing times; shows an inhibitory effect on histamine-induced infiltration of inflammatory cells such as mast cells and eosinophils, and thus has a strong anti-inflammatory and anti-pruritic effect in an atopic dermatitis model; and is selective for the 5-hydroxytryptamine 3 receptor to prevent side effects such as diarrhea or constipation (Clinical and Experimental Immunology, 2010, 161, 19-27; Pharmacology & Therapeutics, 2010, 128, 146-169) due to the high structural similarity between histamine 4 receptor (hH4R) ligands and 5-hydroxytryptamine 3 receptor ligands (BMCL, 2011, 21, 5460-5464). It is therefore an object of the present invention to provide such novel heterocyclic compounds and pharmaceutical compositions comprising them.
Since the novel heterocyclic compounds of the present invention and pharmaceutical compositions comprising them exhibit strong human histamine 4 receptor (hH4R) inhibitory activity, they are useful for the treatment or prevention of inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye, cardiac dysfunction, arrhythmia, atherosclerosis, multiple sclerosis, inflammatory bowel diseases (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritis pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, inflammatory diseases, allergic diseases, and inflammatory diseases, Postoperative adhesions, vestibular disorders, and cancer.
Means for solving the problems
The present invention provides heterocyclic compounds of formula 1
[ formula 1]
Or a racemate, isomer or pharmaceutically acceptable salt thereof:
wherein
X1、X2、X3And X4Each independently is C or N, provided that X1、X2、X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-to 12-membered mono-or poly-heterocyclyl group containing 1 to 3 heteroatoms, preferably a heteroatom selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7、-C1-C6alkyl-NR6R7And R8(ii) a Or R1Selected from-H, -NR6R7And R8，
R2、R3、R4And R5May be the same or different; and each is independently selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -amino-C1-C6An alkyl group; -C3-C8A cycloalkyl group; -halogen (-F, -Cl, -Br, -I); -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8An alkynyl group; -an amino group; -an amido group; -C1-C6An alkylcarboxyl group; -a carboxyl group (-COOH); -C1-C6An acyl group; -OH; -nitro (-NO)2)；-C6-C10An aryl group; -a heterocyclic group; and-O-C1-C6Alkyl-heterocyclyl, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl comprising 1-3 heteroatoms, preferably a heteroatom selected from N, O and S,
with the proviso that when X1When is N, R2Is absent; when X is present2When is N, R3Is absent; when X is present3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1、Y2、Y3、Y4and Y5Each independently being C or a heteroatom (preferably a heteroatom independently selected from N, O and S), with the proviso that Y is1、Y2、Y3、Y4And Y5Is a heteroatom independently selected from N, O and S,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
Y4can be substituted by-H or-C1-C6The substitution of the alkyl group is carried out,
R6and R7Each independently selected from-H; -C1-C6An alkyl group; -C3-C8A cycloalkyl group; -a heterocyclic group; -amino-C1-C6Mono-or di-alkyl; -C1-C6alkyl-amino-C1-C6Mono-or di-alkyl; -C1-C6An alkyl-heterocyclyl group; -C1-C6An alkylcarboxyl group; -a carboxyl group (-COOH); and phenyl, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms, preferably a heteroatom selected from N, O and S,
R8is-C1-C6An alkyl group; -C1-C6An alkoxy group; -OH; -an amino group; -C1-C6An alkyl-amino group; -C3-C8A cycloalkyl group; -S-C1-C6alkyl-amino-C1-C6Mono-or di-alkyl; -S-C1-C6An alkyl-heterocyclyl group; -O-heterocyclyl; or-O-C1-C6Alkyl-heterocyclyl, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl comprising 1-3 heteroatoms, preferably a heteroatom selected from N, O and S, and
R9is selected from-H; -OH; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -amino-C1-C6Mono-or di-alkyl; -C3-C7A cycloalkyl group; -a heterocyclic group; -C6-C10An aryl group; 5-12 membered heteroaryl; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -halogen (-F, -Cl, -Br, -I); -an amino group; -an amido group; -C1-C6An acyl group; -CN; -a carboxyl group (-COOH); -C1-C6An alkylcarboxyl group; and-nitro (-NO)2) Provided that when Y is4Is N and Y1、Y2、Y3And Y5When is C, Y3Is not substituted by a substituent having a-C (═ O) -moiety,
wherein the alkyl, cycloalkyl, heterocyclyl, alkoxy, alkenyl, alkynyl, acyl, and aryl groups each independently may be unsubstituted or substituted with one or more substituents (e.g., 1-3 substituents) selected from-C1-C4Alkyl, -halogen (-F, -Cl, -Br, -I), -CN, -C1-C4Alkoxy, -amino, -amido, -carboxyl (-COOH), -C1-C6Acyl, -OH, -nitro (-NO)2) Heterocyclyl and phenyl, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl comprising 1-3 heteroatoms, preferably a heteroatom selected from N, O and S.
According to a preferred embodiment of the present invention, in the above formula 1, X1、X2And X3Each independently is C or N, and X4Is N.
According to another preferred embodiment of the present invention, in the above formula 1, Y1、Y2、Y3、Y4And Y5Each independently being C or a heteroatom (preferably a heteroatom independently selected from N, O and S), with the proviso that Y is1、Y2、Y3、Y4And Y5Is N.
According to another preferred embodiment of the present invention, in the above formula 1, R1Is a saturated or unsaturated 3-to 8-membered mono-or poly-heterocyclyl group comprising 1 to 3 heteroatoms, preferably a heteroatom selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from NR6R7、C1-C6alkyl-NR6R7And R8。
According to another preferred embodiment of the present invention, in the above formula 1, R3Is selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -halogen (-F, Cl, -Br, -I); -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8Alkynyl and-OH.
According to another preferred embodiment of the present invention, in the above formula 1,
X1、X2、X3and X4Each independently is C or N, provided that X1、X2、X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-to 12-membered mono-or poly-heterocyclyl group containing 1 to 3 heteroatoms, preferably a heteroatom selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7、-C1-C6alkyl-NR6R7And R8(ii) a Or R1Is selected from-NR6R7And R8，
R2、R3、R4And R5May be the same or different; and each is independently selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -halogen (C)-F、-Cl、-Br、-I)；-CN；-C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8An alkynyl group; and an-OH group, and a group,
with the proviso that when X1When is N, R2Is absent; when X is present2When is N, R3Is absent; when X is present3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1、Y2、Y3、Y4and Y5Each independently being C or a heteroatom (preferably a heteroatom independently selected from N, O and S), with the proviso that Y is1、Y2、Y3、Y4And Y5At least two of which are heteroatoms independently selected from N, O and S,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
Y4can be substituted by-H or-C1-C6The substitution of the alkyl group is carried out,
R6and R7Each independently selected from-H; -C1-C6An alkyl group; and-a carboxyl group (-COOH),
R8is selected from-C1-C6An alkyl group; and-C3-C8Cycloalkyl radicals, and
R9is selected from-H; -C1-C6An alkyl group; and-C3-C7A cycloalkyl group,
wherein said alkyl, cycloalkyl, heterocyclyl, alkoxy, alkenyl and alkynyl each independently may be unsubstituted or substituted with one or more substituents selected from-C1-C4Alkyl, -OH and-C1-C4Alkoxy, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms, preferably a heteroatom selected from N, O and S.
According to another preferred embodiment of the present invention, in the above formula 1,
X1、X2、X3and X4Each independently is C or N, provided that X1、X2、X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-to 12-membered mono-or poly-heterocyclyl group containing 1 to 3 heteroatoms, preferably a heteroatom selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7And R8，
R2、R3、R4And R5May be the same or different; and each is independently selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -halogen (-F, -Cl, -Br, -I); -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C2-C7An alkenyl group; and-C2-C8An alkynyl group,
with the proviso that when X1When is N, R2Is absent; when X is present2When is N, R3Is absent; when X is present3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1、Y2、Y3、Y4and Y5Each independently selected from C, N and O, with the proviso that Y1、Y2、Y3、Y4And Y5At least two of which are N or O,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6An alkyl group, a carboxyl group,
R8is selected from-C1-C6Alkyl radical, and
R9is selected from-H, -C1-C6Alkyl and-C3-C7A cycloalkyl group,
wherein said alkyl, cycloalkyl, heterocyclyl, alkoxy, alkenyl and alkynyl each independently may be unsubstituted or substituted with one or more substituents selected from-C1-C4Alkyl, -OH and-C1-C4Alkoxy, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms, preferably a heteroatom selected from N, O and S.
According to another preferred embodiment of the present invention, in the above formula 1,
X1、X2、X3and X4Each independently is C or N, provided that X1、X2、X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-6 membered heterocyclic group containing 1-3 heteroatoms, preferably selected from N, O and S, wherein R is1Unsubstituted or substituted by-NR6R7The substitution is carried out by the following steps,
R2、R3、R4and R5May be the same or different; and each is independently selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -halogen (-F, Cl, -Br, -I); -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C2-C7An alkenyl group; and-C2-C8An alkynyl group,
with the proviso that when X1When is N, R2Is absent; when X is present2When is N, R3Is absent; when X is present3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1、Y2、Y3、Y4and Y5Each independently selected from C, N and O, with the proviso that Y1、Y2、Y3、Y4And Y5At least two of which are N,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6Alkyl radical, and
R9is selected from-H, -C1-C6Alkyl and-C3-C7A cycloalkyl group,
wherein said alkyl, cycloalkyl, heterocyclyl, alkoxy, alkenyl and alkynyl each independently may be unsubstituted or substituted with one or more substituents selected from-C1-C4Alkyl, -OH and-C1-C4Alkoxy, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms, preferably a heteroatom selected from N, O and S.
According to another preferred embodiment of the present invention, in the above formula 1,
X1、X2、X3and X4Each independently is C or N, provided that X1、X2、X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-6 membered heterocyclic group containing 1-3 heteroatoms, preferably selected from N, O and S, wherein R is1Unsubstituted or substituted by-NR6R7The substitution is carried out by the following steps,
R2、R3、R4and R5May be the same or different; and each is independently selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; and-halogen (-F, -Cl, -Br, -I),
with the proviso that when X1When is N, R2Does not storeAt least one of the following steps; when X is present2When is N, R3Is absent; when X is present3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1、Y2、Y3、Y4and Y5Each independently selected from C, N and O, with the proviso that Y1、Y2、Y3、Y4And Y5At least two of which are N,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6Alkyl radical, and
R9is selected from-H and-C1-C6An alkyl group, a carboxyl group,
wherein said alkyl and heterocyclyl may each independently be unsubstituted or substituted by one or more substituents selected from-C1-C4Alkyl, -OH and-C1-C4Alkoxy, wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms, preferably a heteroatom selected from N, O and S.
The compounds of the invention are inhibitors of the human histamine 4 receptor (hH4R) and are useful in the treatment or prevention of inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye, cardiac dysfunction, cardiac arrhythmias, atherosclerosis, multiple sclerosis, inflammatory bowel disease (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, post-operative adhesions, vestibular disorders, and cancer, and in particular as an agent for the treatment of atopic dermatitis.
Unless otherwise noted, the alkyl residues in the alkyl substituents as described herein and in other substituents (e.g., alkoxy) as described herein may be straight or branched chain. Further, the halogen includes fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).
As representative examples of the compound of formula 1 of the present invention, the following compounds may be mentioned:
3-methyl-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 1);
8-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 2);
4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 3);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 4);
3-chloro-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 5);
6- (4-Methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (Compound 6);
8-chloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 7);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 8);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylpyrrolidin-3-amine (compound 9);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine (compound 10);
(R) -1- (3-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylpyrrolidin-3-amine (compound 11);
8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 12);
4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 13);
8-chloro-1-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 14);
8-chloro-1-methyl-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 15);
8-bromo-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 16);
7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 17);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 18);
(S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 19);
(S) -8-chloro-4- (3, 4-dimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 20);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile (compound 21);
8-chloro-4- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 22);
8-chloro-7-ethoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 23);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 24);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 25);
4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 26);
1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 27);
8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile (compound 28);
8-chloro-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 29);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-amine (compound 30);
9-chloro-2-methyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 31);
9-chloro-2-methyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 32);
1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 33);
9-chloro-2-cyclopropyl-N, N-diethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-amine (compound 34);
9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 35);
1- (9-chloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 36);
9-chloro-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 37);
9-chloro-2-cyclopropyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 38);
9-chloro-2-cyclopropyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 39);
9-chloro-2- (methoxymethyl) -5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 40);
9-chloro-2-ethyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 41);
9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-c ] pyrimidine (compound 42);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine (compound 43);
1- (8-chloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine (compound 44);
8-chloro-2-methyl-4- (piperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine (compound 45);
8-chloro-4- (4-methylpiperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine (compound 46);
8-chloro-4- (piperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine (compound 47);
1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 48);
8-iodo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 49);
n-methyl-1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 50);
1- (8- (difluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 51);
n-methyl-1- (8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 52);
4- (4-methylpiperazin-1-yl) -8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 53);
1- (8-ethynylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 54);
n-methyl-1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 55);
1- (8-ethylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 56);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-ol (compound 57);
1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 58);
1- (8- (difluoromethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 59);
8-chloro-7-methoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 60);
8-chloro-7-methoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 61);
7, 8-dichloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 62);
8-chloro-7-ethoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 63);
8-chloro-4- (4-methylpiperazin-1-yl) -7- (2,2, 2-trifluoroethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 64);
1- (8-bromo-9-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 65);
8-bromo-9-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 66);
1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 67);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride (compound 68);
1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride (compound 69);
8-chloro-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 70);
8-bromo-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 71);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-amine (compound 72);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, 3-dimethylazetidin-3-amine (compound 73);
8-bromo-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 74);
4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 75);
8-chloro-4- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 76);
4- ((1S,4S) -2, 5-diazabicyclo [2.2.1] hept-2-yl) -8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 77);
8-chloro-4- ((1S,4S) -5-methyl-2, 5-diazabicyclo [2.2.1] hept-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 78);
8-chloro-4- (1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 79);
8-chloro-4- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 80);
(R) -1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine (compound 81);
8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (Compound 82);
8-chloro-4- (1-methylhexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 83);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (compound 84);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (compound 85);
(1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid (compound 86);
2- ((8-Chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) ethanol (Compound 87);
1- (8-chloroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 88);
1- (8-bromoimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 89);
(tert-butyl 1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate (compound 90);
1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine (compound 91);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) oxazolo [4,5-c ] [1,8] naphthyridine (compound 92);
1- (8-chloropyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 93);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine (compound 94);
1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 95);
8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine (compound 96);
1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 97);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine (compound 98);
1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 99);
8-bromo-7-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 100);
8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-7-ol HCl salt (compound 101);
n- (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine (compound 102);
1- (2-bromopyrido [3,2-e ] pyrrolo [1,2-c ] pyrimidin-6-yl) -N-methylazetidin-3-amine (compound 103);
1- (8-Bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2,2, 2-trifluoroacetate (Compound 104);
(S) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 105);
(R) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 106);
1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 107);
1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 108);
1- (9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 109);
1- (9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 110);
n-methyl-1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 111);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-amine (compound 112);
n-methyl-1- (8-phenylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 113);
1- (8- (furan-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 114);
1- (2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6-yl) -N-methylazetidin-3-amine (compound 115);
1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylazetidin-3-amine (compound 116);
1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 117);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine methanesulfonate (compound 118);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine maleate (compound 119);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2-hydroxypropane-1, 2, 3-tricarboxylate (compound 120);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine nitrate (compound 121);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydroiodide (compound 122);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine phosphate (compound 123);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 4,4' -methylenebis (3-hydroxy-2-naphthoic acid) salt (compound 124);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrobromide (compound 125);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine sulfate (compound 126);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (2R,3R) -2, 3-dihydroxysuccinate salt (compound 127);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (1S) - (+) -10-camphorsulfonate (compound 128);
8-bromo-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine (compound 129);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (S) -2-hydroxypropionate (compound 130);
n- (azetidin-3-ylmethyl) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine trifluoroacetate (compound 131); and
4- (azetidin-3-ylmethoxy) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine hydrochloride (Compound 132).
The names of the compounds listed above are described in accordance with chembidraw Ultra software (version 12.02.1076) by cambridge soft.
In the case where the compound of formula 1 of the present invention is a racemate, the racemate may be separated into its corresponding isomers by using a conventional separation method, such as column chromatography generally filled with normal phase silica gel (Isu Chemical Co., particle diameter: 0.040 to 0.063mm and 0.063 to 0.200mm), column chromatography generally filled with amine silica gel (Isu Chemical Co., particle diameter: 0.040 to 0.075mm), or reverse phase filled pressurized fractionation column chromatography (Yamazen, W-Prep2XY) using corresponding solvents, preferably a solvent mixture of hexane, ethyl acetate, dichloromethane and methanol in normal phase and a solvent mixture of water and acetonitrile in reverse phase.
The compounds of the present invention formula 1 may also form pharmaceutically acceptable salts. Representative acids for use in preparing such pharmaceutically acceptable salts (e.g., acid addition salts) include, but are not limited to, hydrochloric, sulfuric, nitric, phosphoric, hydroiodic, formic, citric, acetic, trichloroacetic or trifluoroacetic acids, benzoic, fumaric, maleic, methanesulfonic, benzenesulfonic, p-toluenesulfonic, 2-dichloroacetic acid, acylated amino acids, adipic, alginic, ascorbic, L-aspartic, 4-acetamidobenzoic, (+) -camphoric, camphorsulfonic, (+) - (1S) -camphorsulfonic, capric, hexanoic, octanoic, cinnamic, cyclamic, dodecylsulfuric, ethane-1, 2-disulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, galactaric, gentisic, glucoheptonic, D-gluconic, acetic, propionic, d-glucuronic acid, L-glutamic acid, alpha-oxo-glutaric acid, glycolic acid, hippuric acid, (+) -L-lactic acid, (+ -) -DL-lactic acid, lactobionic acid, (-) -L-malic acid, malonic acid, (+ -) -DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1, 5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, tannic acid, (+) -L-tartaric acid, thiocyanic acid, undecylenic acid and the like. In addition, other acid salts known and used in the art of amine derivatives may be included. They can be prepared by conventionally known methods.
The compound of formula 1 of the present invention as defined above may be prepared by the methods described in the following embodiments, but is not limited thereto.
The heterocyclic compound having the structure of formula 1 or its racemate, isomer or pharmaceutically acceptable salt may be prepared by a method comprising the steps of:
(a) by using the formula R1-H (except for R)1Is other than-H) arylation of a compound of formula 4 to produce a compound of formula 3An agent;
(b) preparing a compound of formula 2 by arylating the prepared compound of formula 3; and
(c) cyclization of the prepared compound of formula 2 (alternatively, may comprise subjecting R to1Deprotection step):
[ formula 2]
[ formula 3]
[ formula 4]
Wherein in the formulae 2 to 4, X1、X2、X3、X4、R1、R2、R3、R4、R5、R9And Y5The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The heterocyclic compound having the structure of formula 5 or its racemate, isomer or pharmaceutically acceptable salt may be prepared by a method wherein formula 5 is wherein Y1、Y2And Y4Is N, formula 1 as defined above, the process comprising the steps of:
(a) preparing a compound of formula 9 by halogenating a compound of formula 10;
(b) preparing a compound of formula 8 by cyanating the prepared compound of formula 9;
(c) preparing a compound of formula 7 by acylating the prepared compound of formula 8;
(d) preparing a compound of formula 6 by cyclizing the prepared compound of formula 7 followed by halogenation; and
(e) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, may comprise subjecting R to1Deprotection step):
[ formula 5]
[ formula 6]
[ formula 7]
[ formula 8]
[ formula 9]
[ formula 10]
Wherein in formulas 5-10，X1、X2、X3、X4、R1、R2、R3、R4、R5、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group such as halogen (-F, -Cl, -Br), mesylate, triflate, p-tosylate.
The heterocyclic compound having the structure of formula 11, or its racemate, isomer or pharmaceutically acceptable salt thereof, may be prepared by a method wherein formula 11 is wherein Y2、Y4And Y5Is N, formula 1 as defined above, the process comprising the steps of:
(a) preparing a compound of formula 16 by arylating a compound of formula 17;
(b) preparing a compound of formula 15 by imidizing the prepared compound of formula 16;
(c) preparing a compound of formula 14 by acylating the prepared compound of formula 15;
(d) preparing a compound of formula 13 by cyclizing the prepared compound of formula 14;
(e) preparing a compound of formula 12 by reducing the prepared compound of formula 13, followed by halogenation; and
(f) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, may comprise subjecting R to1Deprotection step):
[ formula 11]
[ formula 12]
[ formula 13]
[ formula 14]
[ formula 15]
[ formula 16]
[ formula 17]
Wherein in formulae 11-17, X1、X2、X3、X4、R1、R2、R3、R4、R5、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The heterocyclic compound having the structure of formula 18 or its racemate, isomer or pharmaceutically acceptable salt may be prepared by a method wherein formula 18 is wherein Y2And Y3Is N, formula 1 as defined above, the process comprising the steps of:
(a) preparing a compound of formula 21 by subjecting a compound of formula 23 to Friedel-crafts reaction with a compound of formula 22;
(b) preparing a compound of formula 20 by cyclizing the prepared compound of formula 21;
(c) preparing a compound of formula 19 by halogenating the prepared compound of formula 20; and
(d) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, may comprise subjecting R to1Deprotection step):
[ formula 18]
[ formula 19]
[ formula 20]
[ formula 21]
[ formula 22]
[ formula 23]
Wherein in the formulae 18 to 23, X1、X2、X3、X4、R1、R2、R3、R4、R5And R9The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The heterocyclic compound having the structure of formula 24, or its racemate, isomer or pharmaceutically acceptable salt thereof, can be prepared by a method wherein Y is formula 241、Y3And Y4Is N, formula 1 as defined above, the process comprising the steps of:
(a) preparing a compound of formula 28 by cyclizing a compound of formula 29;
(b) preparing a compound of formula 27 by halogenating the prepared compound of formula 28 followed by hydroxylation;
(c) by using the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 27, followed by halogenation to prepare a compound of formula 26;
(d) preparing a compound of formula 25 by arylating the prepared compound of formula 26; and
(e) cyclization of the prepared compound of formula 25 (alternatively, may comprise subjecting R to1Deprotection step):
[ formula 24]
[ formula 25]
[ formula 26]
[ formula 27]
[ formula 28]
[ formula 29]
Wherein in the formulae 24 to 29, X1、X2、X3、X4、R1、R2、R3、R4、R5And R9The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The heterocyclic compound having the structure of formula 30, or its racemate, isomer or pharmaceutically acceptable salt thereof, may be prepared by a method wherein Y is formula 302Is N, Y4Is O, formula 1 as defined above, comprising the steps of:
(a) preparing a compound of formula 36 by esterifying a compound of formula 37;
(b) preparing a compound of formula 35 by arylating the prepared compound of formula 36;
(c) preparing a compound of formula 34 by cyclizing the prepared compound of formula 35;
(d) preparing a compound of formula 33 by enolization addition reaction of the prepared compound of formula 34;
(e) preparing a compound of formula 32 by cyclizing the prepared compound of formula 33;
(f) preparing a compound of formula 31 by halogenating the prepared compound of formula 32; and
(g) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, can comprise subjecting R to1Deprotection step):
[ formula 30]
[ formula 31]
[ formula 32]
[ formula 33]
[ formula 34]
[ formula 35]
[ formula 36]
[ formula 37]
Wherein in the formulae 30 to 37, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6And R9The same as defined in formula 1; p represents a protecting group such as P-methoxybenzyl, 3',5' -dimethoxybenzyl, tri-methoxybenzyl; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The heterocyclic compound having the structure of formula 38 or its racemate, isomer or pharmaceutically acceptable salt may be prepared by a method wherein said formula 38 is wherein Y1Is N and Y2And Y3Formula 1 as defined above, each independently being C or N, the process comprising the steps of:
(a) preparing a compound of formula 40 by Suzuki (Suzuki) coupling reaction of a compound of formula 42 with a compound of formula 41;
(b) preparing a compound of formula 39 by halogenating the prepared compound of formula 40; and
(c) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, can comprise subjecting R to1Deprotection step):
[ formula 38]
[ formula 39]
[ formula 40]
[ formula 41]
[ formula 42]
Wherein in the formulae 38 to 42, X1、X2、X3、X4、R1、R2、R3、R4And R5The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
A heterocyclic compound having the structure of formula 43 or a racemate, isomer or pharmaceutically acceptable salt thereof, wherein formula 43 is wherein Y5Is N, formula 1 as defined above, the process comprising the steps of:
(a) preparing a compound of formula 46 by pyrrolation of a compound of formula 47;
(b) preparing a compound of formula 45 by cyclizing the prepared compound of formula 46;
(c) preparing a compound of formula 44 by halogenating the prepared compound of formula 45; and
(d) using the formula R1-H (except for R)1Is other than-H) arylating (or, alternatively, can comprise subjecting R to1Deprotection step):
[ formula 43]
[ formula 44]
[ formula 45]
[ formula 46]
[ formula 47]
Wherein in the formulae 43 to 47, X1、X2、X3、X4、R1、R2、R3、R4And R5The same as defined in formula 1; and Z represents a reactive leaving group, such as halogen (-F, -Cl, -Br).
The compound of formula 1 of the present invention has excellent human histamine 4 receptor (hH4R) inhibitory activity. Accordingly, the present invention also provides a pharmaceutical composition comprising an effective amount of the compound of formula 1 or its racemate, isomer or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
The pharmaceutical composition of the present invention may be prepared by mixing an effective amount of the compound of formula 1 or its racemate, isomer or pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier, binder, stabilizer and/or diluent. In addition, when preparing the pharmaceutical composition of the present invention in the form of an injectable liquid, a pharmaceutically acceptable buffer, a dissolution adjuvant and/or an isotonic agent may be mixed with the compound of formula 1 or its racemate, isomer or pharmaceutically acceptable salt.
Since the pharmaceutical composition of the present invention shows strong human histamine 4 receptor (hH4R) inhibitory activity, it is useful for treating or preventing inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye, cardiac dysfunction, cardiac arrhythmias, atherosclerosis, multiple sclerosis, inflammatory bowel disease (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, post-operative adhesions, vestibular disorders, and cancer.
The pharmaceutical compositions of the present invention may be prepared as pharmaceutical compositions containing one or more unit doses of the pharmaceutically active agent by using manufacturing techniques known or available in the art, suitable pharmaceutical excipients. In the methods of the invention, the composition may be administered by a suitable delivery route, for example oral or parenteral, transdermal, rectal, topical or ocular administration or by inhalation. The pharmaceutical preparations can be in the form of tablets, capsules, sachets, dragees, powders, granules, lozenges, redissolved powders, liquid preparations or suppositories. For example, the composition may be formulated for intravenous injection, spray, topical or oral administration.
In the case of preparing a formulation for oral dosage form, any conventional pharmaceutical carrier may be employed. For example, water, glycols, oils, alcohols, and the like may be used as carriers in the case of oral liquid preparations, such as suspensions, syrups, elixirs, and solutions; starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like may be used as carriers in the case of solid formulations, such as powders, pills, capsules and tablets. Tablets and capsules are the most convenient dosage form because of the convenience of administration, and are preferably prepared as enteric-coated preparations.
In the case of parenteral formulations, sterile water is typically used, and other ingredients, such as dissolution aids, may also be included. For example, sterile aqueous or oily suspensions for injection may be prepared according to the known art by using suitable dispersing, wetting or suspending agents. Solvents used for this purpose include water, ringer's solution and isotonic NaCl solution, usually with sterile, fixed oils being also used as solvent or suspending medium. Any non-irritating fixed oil including mono-and diglycerides can be used for this purpose, and fatty acids such as oleic acid can be used for the injections.
In the case of transdermal preparations, penetration enhancers and/or suitable humectants can be used as carriers, optionally in combination with suitable additives that are non-irritating to the skin. As such additives, those may be selected that help facilitate administration through the skin and/or preparation of the desired composition. Transdermal formulations may be administered by different means, such as transdermal patches, drops or ointments.
The administration time and dose of the pharmaceutical composition of the present invention can be appropriately determined according to the disease, condition, age, body weight and administration form of the patient. In the case of adults, the pharmaceutical composition is administered in an amount of 0.1 to 2,000mg, preferably 1 to 200 mg/day, in a single dose or multiple doses, but is not limited thereto.
Since the pharmaceutical composition of the present invention shows strong human histamine 4 receptor (hH4R) inhibitory activity, it is useful for treating or preventing inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye, cardiac dysfunction, cardiac arrhythmias, atherosclerosis, multiple sclerosis, inflammatory bowel disease (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, post-operative adhesions, vestibular disorders, and cancer.
Unless otherwise described, the alkyl moieties described herein and the alkyl moieties of other groups (e.g., alkoxy) can be of a straight or branched chain variety. Further, halogens include fluorine, chlorine, bromine and iodine.
The compound of formula 1 of the present invention may be a racemate. The racemate may be separated into its corresponding isomers by using a conventional separation method such as normal phase column chromatography packed with normal phase silica gel (Merck, 0.040 to 0.063mm and 0.063 to 0.200mm), normal phase column chromatography packed with amine silica gel (chromotrex, 100 to 200 mesh) or preparative pressure reverse phase column chromatography (Yonglin, SDV30+), in which respective solvents are used, preferably a solvent mixture of hexane, ethyl acetate, dichloromethane and methanol in the normal phase and a solvent mixture of water and acetonitrile in the reverse phase.
The compounds of formula 1 of the present invention may also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts include: salts derived from the addition of acids which form non-toxic addition salts containing pharmaceutically acceptable anions-e.g., inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydrogen iodide; organic acids, such as tartaric, formic, citric, acetic, trichloroacetic or trifluoroacetic acid, gluconic, benzoic, lactic, fumaric, maleic, sulfonic acids, such as methanesulfonic, benzenesulfonic, p-toluenesulfonic, naphthalenesulfonic acid; and salts of alkali metals such as sodium, potassium, and the like. In addition, acid or base salts known and used in the art of aromatic amidine derivatives and lactam derivatives may be included. The salts may be prepared using conventional methods.
The compound of formula 1 of the present invention can be prepared by the following method, and therefore, the present invention also provides a method for preparing the compound of formula 1.
More specifically, the compound of formula 1 can be prepared by the methods 1 to 8 each described hereinafter, but is not limited thereto.
Method 1
The compound having the structure of formula 2 below, said formula 2 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the following steps2And Y5Is N: by using the formula R1-H (except for R)1Is other than-H) arylating a compound of formula 4 to produce a compound of formula 3; preparing a compound of formula 2 by arylating the prepared compound of formula 3; and cyclizing the prepared compound of formula 2:
[ formula 2]
[ formula 3]
[ formula 4]
Wherein in formulaIn 2-4, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 1 is described in more detail below.
Compounds of formulae 2,3 and 4 which can be used as starting materials can be prepared according to methods well known in the art (e.g., J.Med.chem.1990, 33, 2240-.
By using the formula R1-H (except for R)1Is other than-H) arylating the compound of formula 4 to prepare the compound of formula 3 is carried out according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is dichloromethane.
The next step of preparing the compound of formula 2 by arylating the prepared compound of formula 3 is carried out according to a conventional method in the presence of a suitable solvent.
In general, conventional solvents which do not adversely affect the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The next step of preparing the compound of formula 1 by cyclizing the prepared compound of formula 2 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The cyclization reaction is carried out at 50-200 deg.C for 0.1-24 hr, preferably at 80 deg.C for 1 hr using trimethyl orthoformate or triethyl orthoformate.
The preparation of the above compound of formula 2 is more specifically described in the examples below.
Method 2
Can be prepared by a method comprising the following stepsA compound having the structure of the following formula 5, wherein the following formula 5 is formula 1 as defined above, wherein Y1、Y2And Y4Is N: preparing a compound of formula 9 by halogenating a compound of formula 10 below; preparing a compound of formula 8 by cyanating the prepared compound of formula 9; preparing a compound of formula 7 by acylating the prepared compound of formula 8; preparing a compound of formula 6 by cyclizing the prepared compound of formula 7 followed by halogenation; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 6:
[ formula 5]
[ formula 6]
[ formula 7]
[ formula 8]
[ formula 9]
[ formula 10]
Wherein in formulae 5-10, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8、R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br), a mesylate, a triflate and a p-toluenesulfonate.
Method 2 is described in more detail below.
The compounds of formulae 5,6, 7,8, 9 and 10 used as starting materials can be prepared according to methods well known in the art.
Preparation of the compound of formula 9 by halogenation of the compound of formula 10 is carried out according to conventional methods in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is chloroform.
Typical halogenating agents for use in the reaction include bromine, phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Bromine is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 8 by cyanating the prepared compound of formula 9 is carried out according to a conventional method in the presence of a suitable solvent and a cyanating agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N-methylpyrrolidone.
Typical cyanating agents for the reaction include KCN, NaCN, Zn (CN)2、CuCN、(CH3)2C (OH) CN and TMSCN. Preference is given to Zn (CN)2. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 7 by acylating the prepared compound of formula 8 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is 2-butanone.
The next step of preparing the compound of formula 6 by cyclizing the prepared compound of formula 7 followed by halogenation is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is diphenyl ether.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
By using the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 6 to prepare the compound of formula 5 according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N, N-dimethylformamide.
The preparation of the compound of formula 5 above is more particularly described in the following examples.
Method 3
The compound having the structure of formula 11 below, said formula 11 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of2、Y4And Y5Is N: preparing a compound of formula 16 by arylating a compound of formula 17; preparing a compound of formula 15 by imidizing the prepared compound of formula 16; preparing a compound of formula 14 by acylating the prepared compound of formula 15; preparing a compound of formula 13 by cyclizing the prepared compound of formula 14; preparing a compound of formula 12 by reducing the prepared compound of formula 13, followed by halogenation; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 12:
[ formula 11]
[ formula 12]
[ formula 13]
[ formula 14]
[ formula 15]
[ formula 16]
[ formula 17]
Wherein in formulae 11-17, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8、R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 3 is described in more detail below.
The compounds of formulae 11, 12, 13, 14, 15, 16 and 17 used as starting materials can be prepared according to methods well known in the art.
The preparation of the compound of formula 16 by arylating the compound of formula 17 is carried out according to conventional methods in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The next step of preparing the compound of formula 15 by imidating the prepared compound of formula 16 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is pyridine.
The next step of preparing the compound of formula 14 by acylating the prepared compound of formula 15 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is diethyl ether.
The next step of preparing the compound of formula 13 by cyclizing the prepared compound of formula 14 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is toluene.
The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 12 by reducing the prepared compound of formula 13 and then halogenating is carried out according to a conventional method in the presence of a suitable solvent, a reducing agent and a halogenating agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is acetic acid.
Typical reducing agents for this reaction include palladium on carbon catalyst (5% w/w), palladium on carbon catalyst (10% w/w), Raney nickel, zinc and iron, with iron being particularly preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
In addition, typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride, and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
By using the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 12 to prepare the compound of formula 11 according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N, N-dimethylformamide.
The preparation of the above compound of formula 11 is more specifically described in the following examples.
Method 4
The compound having the structure of formula 18 below, said formula 18 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of2And Y3Is N: preparing a compound of formula 21 below by friedel-crafts reaction of a compound of formula 23 below with a compound of formula 22 below; preparing a compound of formula 20 by cyclizing the prepared compound of formula 21; preparing a compound of formula 19 by halogenating the prepared compound of formula 20; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 19:
[ formula 18]
[ formula 19]
[ formula 20]
[ formula 21]
[ formula 22]
[ formula 23]
Wherein in the formulae 18 to 23, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8、R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 4 is described in more detail below.
The compounds of formulae 18, 19, 20, 21, 22 and 23 used as starting materials can be prepared according to methods well known in the art.
The preparation of the compound of formula 21 by friedel-crafts reaction of the compound of formula 23 with the compound of formula 22 is carried out according to conventional methods in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is dichloromethane.
The next step of preparing a compound of the following formula 20 by cyclizing the prepared compound of the formula 21 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 19 by halogenating the prepared compound of formula 20 is carried out according to a conventional method in the presence of a suitable solvent and halogenating agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is toluene. No solvent may be used in the reaction.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
By using the formula R1-H (except for R)1Is other than-H) arylation of the prepared compound of formula 19 to prepare the compound of formula 18 arylation is carried out according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N, N-dimethylformamide.
The preparation of the above compound of formula 18 is more specifically described in the following examples.
Method 5
The compound having the structure of formula 24 below, said formula 24 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of1、Y3And Y4Is N: preparing a compound of formula 28 below by cyclizing a compound of formula 29 below; preparing a compound of formula 27 by halogenating the prepared compound of formula 28 followed by hydroxylation; by using the formula R1-H (except for R)1Is other than-H) so as to prepareArylating the compound of formula 27, followed by halogenation to produce a compound of formula 26; preparing a compound of formula 25 by arylating the prepared compound of formula 26; and cyclizing the prepared compound of formula 25:
[ formula 24]
[ formula 25]
[ formula 26]
[ formula 27]
[ formula 28]
[ formula 29]
Wherein in the formulae 24 to 29, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8And R9And defined in formula 1The same; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 5 is described in more detail below.
The compounds of formulae 24, 25, 26, 27, 28 and 29 used as starting materials can be prepared according to methods well known in the art.
Preparation of a compound of formula 28 by cyclization of a compound of formula 29 is carried out according to conventional methods in the presence of a suitable solvent and a cyclizing agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is dioxane.
Typical cyclizing agents used in this reaction include diphosgene and triphosgene. Diphosgene is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing a compound of formula 27 by halogenation of the prepared compound of formula 28 followed by hydroxylation is carried out according to conventional methods in the presence of suitable solvents, halogenating agents and hydroxylating agents.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is n-butanol.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
Typical hydroxylating agents used in the reaction include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide and magnesium hydroxide. Sodium hydroxide is preferred. The reaction may be carried out at room temperature for about 0.1 to 24 hours, but is not limited thereto.
By using the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 27, and then halogenating to prepare the compound of formula 26 according to a conventional method in the presence of a suitable solvent and halogenating agent.
Conventional solvents which do not adversely affect the reaction can be used for the arylation reaction. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 25 by arylating the prepared compound of formula 26 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The next step of preparing a compound of formula 24 by cyclizing the prepared compound of formula 25 is carried out according to conventional methods in the presence of a suitable solvent and a cyclizing agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
Typical preferred cyclizing agents for use in this reaction include trimethyl orthoformate and triethyl orthoformate. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The preparation of the above compound of formula 24 is more specifically described in the following examples.
Method 6
The compound having the structure of formula 30 below, said formula 30 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of2Is N and Y4Is O: preparing a compound of formula 36 below by esterifying a compound of formula 37 below; preparing a compound of formula 35 by arylating the prepared compound of formula 36; preparing a compound of formula 34 by cyclizing the prepared compound of formula 35; preparing a compound of formula 33 by enolization addition reaction of the prepared compound of formula 34; preparing a compound of formula 32 by cyclizing the prepared compound of formula 33; preparing a compound of formula 31 by halogenating the prepared compound of formula 32; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 31:
[ formula 30]
[ formula 31]
[ formula 32]
[ formula 33]
[ formula 34]
[ formula 35]
[ formula 36]
[ formula 37]
Wherein in the formulae 30 to 37, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8And R9The same as defined in formula 1; z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br); and P represents a protecting group, preferably P-methoxybenzyl, 3',5' -dimethoxybenzyl and tri-methoxybenzyl.
Method 6 is described in more detail below.
The compounds of formulae 30, 31, 32, 33, 34, 35, 36 and 37 used as starting materials can be prepared according to methods well known in the art.
Preparation of a compound of formula 36 by esterification of a compound of formula 37 below was carried out according to a conventional method in the presence of a suitable solvent and an esterification reagent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. Particularly preferred solvents for this reaction are dichloromethane and methanol.
The next step of preparing the compound of formula 35 by arylating the prepared compound of formula 36 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is ethanol.
The next step of preparing a compound of formula 34 by cyclizing the prepared compound of formula 35 is carried out according to a conventional method in the presence of a suitable solvent and a cyclizing agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is dioxane.
Typical cyclizing agents used in this reaction include diphosgene and triphosgene. Diphosgene is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 33 by enolization addition reaction of the prepared compound of formula 34 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is tetrahydrofuran.
The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 32 by cyclizing the prepared compound of formula 33 is carried out according to a conventional method in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is acetic acid.
The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing the compound of formula 31 by halogenating the prepared compound of formula 32 is carried out according to conventional methods in the presence of a suitable halogenating agent.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step using the formula R1-H (except for R)1Is other than-H) arylation of the prepared compound of formula 31 to prepare a compound of formula 30According to conventional methods in a suitable solvent and alkali presence.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N, N-dimethylformamide.
The preparation of the compound of formula 30 above is more particularly described in the following examples.
Method 7
The compound having the structure of formula 38 below, said formula 38 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of1Is N, Y2And Y3Each independently is C or N and Y1、Y2And Y3Is N: preparing a compound of formula 40 below by suzuki coupling reaction of a compound of formula 42 below with a compound of formula 41 below; preparing a compound of formula 39 by halogenating the prepared compound of formula 40; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 39:
[ formula 38]
[ formula 39]
[ formula 40]
[ formula 41]
[ formula 42]
Wherein in the formulae 38 to 42, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 7 is described in more detail below.
The compounds of formulae 38, 39, 40, 41 and 42 used as starting materials can be prepared according to methods well known in the art.
Preparation of the compound of formula 40 by suzuki coupling reaction of the compound of formula 42 with the compound of formula 41 is carried out according to a conventional method in the presence of a suitable solvent and catalyst.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents, e.g. benzene, pyridine,
Toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents, e.g. dichloromethane, chloroform, methanol, ethanol,
carbon tetrachloride, 1, 2-dichloroethane, and the like; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents, e.g. acetone, N-dimethylformamide, N-dimethylacetamide,
N-methylpyrrolidone, dimethylsulfoxide, acetonitrile, etc.; and mixed solvents thereof. Particularly preferred solvents for the reaction are N, N-dimethylformamide and water.
Typical catalysts for the reaction include tetrakis (triphenylphosphine) palladium (Pd (PPh)3)4) Palladium (II) acetate (Pd (OAc)2) Bis (triphenylphosphine) palladium (II) dichloride (PdCl)2(PPh3)2) [1, 1-bis (diphenylphosphino) ferrocene]dichloro-Palladium (II) (PdCl)2(dppf)), (dibenzylideneacetone) dipalladium (0) (Pd (dba)2) And palladium (II) chloride (PdCl)2). Tetrakis (triphenylphosphine) palladium (Pd (PPh) is preferred3)4). The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing a compound of formula 39 by halogenating the prepared compound of formula 40 is carried out according to conventional methods in the presence of a suitable halogenating agent.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step using the formula R1-H (except for R)1Is other than-H) arylation of the prepared compound of formula 39 to prepare the compound of formula 38 is carried out according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is dichloromethane.
The preparation of the above compound of formula 5 is more specifically described in the following examples.
Method 8
The compound having the structure of formula 43 below, said formula 43 below being formula 1 as defined above, wherein Y may be prepared by a method comprising the steps of5Is N: preparing a compound of formula 46 below by pyrrolation of a compound of formula 47 below; preparing a compound of formula 45 by cyclizing the prepared compound of formula 46; preparing a compound of formula 44 by halogenating the prepared compound of formula 45; and use of the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 44:
[ formula 43]
[ formula 44]
[ formula 45]
[ formula 46]
[ formula 47]
Wherein in the formulae 43 to 47, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6、R7、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group, preferably a halogen atom (-F, -Cl, -Br).
Method 8 is described in more detail below.
The compounds of formulae 43, 44, 45, 46 and 47 used as starting materials can be prepared according to methods well known in the art.
Preparation of the compound of formula 46 by pyrrolation of the compound of formula 47 is carried out according to conventional methods in the presence of a suitable solvent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is acetic acid. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step of preparing a compound of formula 45 by cyclizing the prepared compound of formula 46 is carried out according to conventional methods in the presence of a suitable solvent and a cyclizing agent.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is toluene.
Typical cyclizing agents used in this reaction include diphosgene and triphosgene. Triphosgene is preferred. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next conventional procedure for preparing formula 44 by halogenating the prepared compound of formula 45 is carried out in the presence of a suitable halogenating agent.
Typical halogenating agents for use in the reaction include phosphorus trichloride, phosphorus oxychloride, phosphorus oxybromide, phenylphosphonic chloride and phosphorus pentachloride. Phosphorus oxychloride is preferred. The reaction may be carried out at 50 to 200 ℃ for about 0.1 to 24 hours, but is not limited thereto.
The next step using the formula R1-H (except for R)1Is other than-H) arylating the prepared compound of formula 44 to prepare the compound of formula 43 according to a conventional method in the presence of a suitable solvent and a base.
Conventional solvents having no adverse effect on the reaction can be used for this step. Preferred examples of the solvent include ether-based solvents such as diethyl ether, tetrahydrofuran, dioxane, 1, 2-dimethoxyethane, 2-butanone, diglyme and the like; hydrocarbon-based solvents such as benzene, pyridine, toluene, hexane, xylene, etc.; halogenated hydrocarbon-based solvents such as dichloromethane, chloroform, carbon tetrachloride, 1, 2-dichloroethane, etc.; alcohol-based solvents such as methanol, ethanol, isopropanol, tert-butanol, and the like; ester-based solvents such as ethyl acetate, methyl acetate, butyl acetate, and the like; polar solvents such as acetone, N-dimethylformamide, N-dimethylacetamide, N-methylpyrrolidone, dimethylsulfoxide, acetonitrile and the like; and mixed solvents thereof. A particularly preferred reaction solvent is N, N-dimethylacetamide.
Advantageous effects of the invention
The novel heterocyclic compounds of the present invention exhibit the same or greater histamine 4 receptor inhibitor activity as compared to commonly used human histamine 4 receptor (hH4R) inhibitors; exhibit selectivity for histamine subtype receptors and receptors, respectively for transporters and for ion channels on membranes; and has higher solubility, metabolic stability, and thus, as a result of pharmacokinetic analysis by using animal models such as SD rats and comparison with the compound disclosed in WO2010/030785, the compound of the present invention shows superior effects in terms of pharmacokinetic properties, such as AUCinfAnd the maximum blood concentration is much more than 7-8 times higher than that of the comparative compound. In terms of symptoms, it was found that itch induced by histamine, substance P, compound 48/80, and the like was effectively suppressed, that is, the compound of the present invention had an excellent effect of being much more than 3 times higher than the compound disclosed in WO2010/030785 in terms of suppressing infiltration of inflammatory cells such as mast cells and eosinophils induced by histamine. In the oxazolone-induced atopic dermatitis model, the present inventionThe heterocyclic compounds of (a) show a far stronger anti-inflammatory action than the compounds disclosed in WO 2010/030785. Especially in a dust mite (dermophagoids farina) induced NC/Nga mouse atopic dermatitis model, the heterocyclic compound of the present invention was found to show an anti-inflammatory effect much higher than that of the immunosuppressive drug tacrolimus, and a compound selective for the 5-hydroxytryptamine 3 receptor was found.
Therefore, the novel heterocyclic compounds of the present invention and the pharmaceutical compositions comprising them can be highly effectively used for the treatment or prevention of inflammatory diseases, allergic reactions, pain, nasal polyps, rhinitis, chronic sinusitis, nasal congestion, nasal pruritus, asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye, cardiac dysfunction, cardiac arrhythmias, atherosclerosis, multiple sclerosis, inflammatory bowel disease (including colitis, crohn's disease, ulcerative colitis), inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated (also referred to as type I) diabetes, lupus, post-operative adhesions, vestibular disorders, and cancer.
Modes for carrying out the invention
The present invention is explained in further detail with reference to the following examples and experiments. However, these examples and experiments are only for illustrating the present invention, and the present invention is not limited thereto in any way.
Abbreviations used in the following examples are defined as follows.
Example 1
Synthesis of 3-methyl-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2-chloro-3-hydrazino-7-methylpyrido [2,3-b ] pyrazine
2, 3-dichloro-7-methylpyrido [2,3-b ]]Pyrazine (500.0mg, 2.33mmol) and hydrazine monohydrate (234.0mg, 4.66mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 12 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid. The resulting solid was then filtered and dried under reduced pressure to give 2-chloro-3-hydrazino-7-methylpyrido [2,3-b ] as a yellow solid]A pyrazine.
LC/MS ESI(+)：210(M+1)，212(M+3)
(b) Synthesis of 6-chloro-3-methylpyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 2-chloro-3-hydrazino-7-methylpyrido [2,3-b ]]A mixture of pyrazine and trimethyl orthoformate (5.0mL) was stirred at 100 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid. Then filtering the formed solid, drying under reduced pressure to obtain milky white solid compound 6-chloro-3-methylpyrido [3,2-e][1,2,4]Triazolo [4,3-a]Pyrazine (340.0mg, 66%, 2 steps).
LC/MS ESI(+)：220(M+1)，222(M+3)
(c) Synthesis of 3-methyl-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
6-chloro-3-methylpyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (50.0mg, 0.23mmol) was dissolved in DMF (2.0mL), to which N-methylpiperazine (48.0mg, 0.48mmol) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour and distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100:0-DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to yield the compound 3-methyl-6- (4-methyl-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a white solid (25.0mg, 38%).
LC/MS ESI(+)：284(M+1)，286(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.83(s，1H)，8.20(m，1H)，7.82(m，1H)，4.34(bs，4H)，2.50(m，4H)，2.42(s，3H)，2.24(s，3H)
Example 2
Synthesis of 8-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2-chloro-7-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2, 3-dichloro-7-methylpyrido [2,3-b ]]Pyrazine (500.0mg, 2.30mmol) was dissolved in DCM (8.0mL) and N-methylpiperazine (468.0mg, 2.60mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, poured into water and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100:0-99: 1). Collecting the product-containing fractions, and evaporating to obtain 2-chloro-7-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as yellow solid]A pyrazine.
LC/MS ESI(+)：278(M+1)，280(M+3)
(b) Synthesis of 8-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 2-chloro-7-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (300.0mg, 1.08mmol) and hydrazine monohydrate (108.0mg, 2.16mmol) were dissolved in EtOH (10.0mL), stirred at 50 ℃ for 12 hours, and then distilled under reduced pressure. The reaction mixture was poured into water and extracted with DCM. The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and evaporating under reduced pressure to obtain yellow solid compound 2-hydrazino-7-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A pyrazine. Unpurified 2-hydrazino-7-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (2.0mL) was stirred at 100 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid and filtering the formed solid. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). Collecting the product-containing fractions, and evaporating to obtain milky white solid compound 8-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (94.0mg, 31%).
LC/MS ESI(+)：284(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.99(s，1H)，8.45(m，2H)，4.37(bs，4H)，2.50(m，4H)，2.43(s，3H)，2.24(s，3H)
Example 3
Synthesis of 4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2, 3-dichloropyrido [2,3-b ]]Pyrazine (300.0mg, 1.50mmol) was dissolved in DMF (5.0mL) and N-methylpiperazine (0.3mL, 2.97mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM)MeOH: 100: 0). Collecting the product-containing fractions, and evaporating to obtain 2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a pale yellow solid]Pyrazine (265.0mg, 67%).
LC/MS ESI(+)：264(M+1)，266(M+3)
(b) Synthesis of 4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (260.0mg, 0.99mmol) and hydrazine monohydrate (63.0mg, 1.97mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and evaporating under reduced pressure to obtain yellow solid compound 2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A pyrazine. Unpurified 2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions to obtain 4- (4-methylpiperazin-1-yl) pyrido [2,3-e as a pale yellow solid][1,2,4]Triazolo [4,3-a]Pyrazine (186.0mg, 70%).
LC/MS ESI(+)：270(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.04(s，1H)，8.61-8.54(m，2H)，7.39-7.32(m，1H)，4.60-4.21(m，4H)，2.55-2.44(m，4H)，2.25(s，3H)
Example 4
Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2,3, 7-trichloropyrido [2,3-b ] pyrazine
5-chloropyridine-2, 3-diamine (10000.0mg, 69.65mmol) was added to diethyl oxalate (30.0mL), and the reaction mixture was stirred at 100 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid. The resulting solid was then filtered and dried under reduced pressure to give 7-chloropyrido [2,3-b ] as a light brown solid]Pyrazine-2, 3-diols. Unpurified 7-chloropyrido [2,3-b ]]Pyrazine-2, 3-diols and POCl3(30.0mL) of the mixture was stirred at 130 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water. The solid formed was filtered and then dried under reduced pressure to give 2,3, 7-trichloropyrido [2,3-b ] as a brown solid]Pyrazine (13700.0mg, 84%, 2 steps).
LC/MS ESI(+)：234(M+1)，236(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.23(d，1H，J＝2.6Hz)，8.86(d，1H，J＝2.6Hz)
(b) Synthesis of 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (428.0mg, 1.83mmol) and TEA (2.5mL, 18.30mmol) were dissolved in DCM (10.0mL) and N-methylpiperazine (0.2mL, 2.01mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and then poured into saturated NH4Aqueous Cl, diluted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]Pyrazine (290.0mg, 53%).
LC/MS ESI(+)：298(M+1)，300(M+3)
(c) Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (290.0mg, 0.97mmol) and hydrazine monohydrate (98.0mg, 1.96mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and then evaporating under reduced pressure to obtain the compound 7-chloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]A pyrazine. Unpurified 7-chloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (5.0mL) was stirred at 80 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (215.0mg, 73%, 2 steps).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，8.85(d，1H，J＝2.4Hz)，8.58(d，1H，J＝2.4Hz)，4.40(bs，4H)，2.53(m，4H)，2.24(s，3H)
Example 5
Synthesis of 3-chloro-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3-hydrazinopyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (150.0mg, 0.64mmol), TEA (129.0mg, 1.28mmol), hydrazine monohydrate (35.0mg, 0.74mmol) were dissolved in EtOH (5.0mL), stirred at room temperature for 12 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid. The solid formed was filtered and then dried under reduced pressure to give 2, 7-dichloro-3-hydrazinopyrido [2,3-b ] as a yellow solid]A pyrazine.
LC/MS ESI(+)：230(M+1)，232(M+3)
(b) Synthesis of 3-chloro-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 2, 7-dichloro-3-hydrazinopyrido [2,3-b ]]A mixture of pyrazine and trimethyl orthoformate (2.0mL) was stirred at 100 ℃ for 2 hours and then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered to give 3, 6-dichloropyrido [3,2-e ] as a yellow solid][1,2,4]Triazolo [4,3-a]A pyrazine. Unpurified 3, 6-dichloropyrido [3,2-e ]][1,2,4]Triazolo [4,3-a]Pyrazine was dissolved in DMF (2.0mL) and N-methylpiperazine (0.1mL, 0.90mmol) diluted with DMF (1.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, poured into water and extracted with EtOAc (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 3-chloro-6- (4-methylpiperazin-1-yl) pyrido [3,2-e][1,2,4]Triazolo [4,3-a]Pyrazine (24.0mg, 12%, 3 steps).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：9.46(s，1H)，8.22(d，1H，J＝2.4Hz)，7.93(d，1H，J＝2.4Hz)，4.54(bs，4H)，2.61(m，4H)，2.37(s，3H)
Example 6
Synthesis of 6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
2, 3-dichloropyrido [2,3-b ]]Pyrazine (300.0mg, 1.50mmol) and hydrazine monohydrate (48.1mg, 1.50mmol) were dissolved in EtOH (7.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and then evaporating under reduced pressure to obtain 2-chloro-3-hydrazinopyrido [2,3-b ]]A pyrazine. Unpurified 2-chloro-3-hydrazinopyrido [2,3-b ]]A mixture of pyrazine and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered and then dried under reduced pressure to give 6-chloropyrido [3,2-e][1,2,4]Triazolo [4,3-a]A pyrazine. Unpurified 6-chloropyrido [3,2-e ]][1,2,4]Triazolo [4,3-a]Pyrazine was dissolved in DMF (5.0mL) and N-methylpiperazine (0.3mL, 2.97mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and then poured into saturated NH4Aqueous Cl, extracted with DCM. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 6- (4-methylpiperazin-1-yl) pyrido [3,2-e as pale yellow solid][1,2,4]Triazolo [4,3-a]Pyrazine (70.0mg, 17%, 3 steps).
LC/MS ESI(+)：270(M+1)
1H-NMR(300MHz，CDCl3)；：9.51(s，1H)，8.30(dd，1H，J＝1.5Hz，J＝4.6Hz)，7.94(dd，1H，J＝1.5Hz，J＝8.0Hz)，7.44(dd，1H，J＝4.6Hz，J＝8.0Hz)，4.70-4.34(m，4H)，2.58-2.56(m，4H)，2.38(s，3H)
Example 7
Synthesis of 8-chloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (800.0mg, 3.41mmol) and TEA (2.4mL, 17.05mmol) were dissolved in DCM (20.0mL) and added slowly thereto at 0 deg.CTert-butyl piperazine-1-carboxylate (667.0mg, 3.58mmol) diluted with DCM (10.0mL) was added. The reaction mixture was stirred at room temperature for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (50.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (n-Hex: EtOAc ═ 90:10-80: 20). Collecting the product-containing fractions, and evaporating to obtain 4- (2, 7-dichloropyrido [2,3-b ] as yellow solid]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester (892.0mg, 68%).
LC/MS ESI(+)：384(M+1)，389(M+3)
(b) Synthesis of tert-butyl 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
4- (2, 7-dichloropyrido [2, 3-b)]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester (892.0mg, 2.32mmol) and hydrazine monohydrate (244.0mg, 4.87mmol) were dissolved in EtOH (50.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (50.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and drying under reduced pressure to obtain the compound 4- (7-chloro-2-hydrazinopyrido [2, 3-b) as yellow solid]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：380(M+1)，382(M+3)
(c) Synthesis of tert-butyl 4- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate
Unpurified 4- (7-chloro-2-hydrazinopyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) piperazine-1-carboxylate and trimethyl orthoformate (20.0mL) was stirred at 70 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered and then dried under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 4- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) piperazinesTert-butyl oxazine-1-carboxylate (814.0mg, 90%).
LC/MS ESI(+)：390(M+1)，392(M+3)
(d) Synthesis of 8-chloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
4- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Tert-butyl pyrazin-4-yl) piperazine-1-carboxylate (814.0mg, 2.09mmol) was dissolved in DCM (20.0mL) and TFA (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (50.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 90:10-80: 20). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 8-chloro-4- (piperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (503.0mg, 83%).
LC/MS ESI(+)：290(M+1)，292(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.00(s，1H)，8.84(d，1H，J＝2.4Hz)，8.56(d，1H，J＝2.4Hz)，4.40(bs，4H)，2.88(m，4H)
Example 8
Synthesis of 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
(a) Synthesis of tert-butyl (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) carbamate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.85mmol) and TEA (1.2mL, 8.50mmol) were dissolved in DCM (10.0mL) and azetidin-3-ylcarbamic acid tert-butyl ester (162.0mg, 0.94mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and poured to saturationNH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 100:0 to 99: 1). The product-containing fractions were collected and evaporated to give (1- (2, 7-dichloropyrido [2, 3-b) as a brown solid]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester (224.0mg, 71%).
LC/MS ESI(+)：370(M+1)，372(M+3)
(b) Synthesis of tert-butyl (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) carbamate
1- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester (224.0mg, 0.61mmol) and hydrazine monohydrate (76.0mg, 1.53mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid, filtering the solid formed, and then drying under reduced pressure to obtain the compound (1- (7-chloro-2-hydrazinopyrido [2, 3-b) as a yellow solid]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester (100.0mg, 41%).
LC/MS ESI(+)：366(M+1)，368(M+3)
(c) Synthesis of tert-butyl (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate
Reacting (1- (7-chloro-2-hydrazinopyrido [2,3-b ]]A mixture of pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester (100.0mg, 0.27mmol) and trimethyl orthoformate (5.0mL) was stirred at 70 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure to obtain a milky white solid compound (1- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (70.0mg, 68%).
LC/MS ESI(+)：376(M+1)，378(M+3)
(d) Synthesis of 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
1- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (70.0mg, 0.19mmol) was dissolved in DCM (5.0mL) to which TFA (2.0mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 90:10 to 50: 50). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-amine (6.3mg, 12%).
LC/MS ESI(+)：276(M+1)，278(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.80(d，1H，J＝2.3Hz)，8.53(d，1H，J＝2.4Hz)，4.93(m，1H)，4.47(m，1H)，4.35(m，1H)，3.93(m，2H)
Example 9
Synthesis of (R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylpyrrolidin-3-amine
Reacting (R) -1- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-amine (20.0mg, 0.07mmol) and sodium formate (20.0mg, 0.29mmol) were dissolved in formic acid/formamide (1.0mL/1.0mL), stirred at 100 ℃ for 1 hour, and then distilled under reduced pressure. With NaHCO3The reaction mixture was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 90:10-80: 20). Collecting the product-containing fraction, evaporating to obtainMilky white solid compound (R) -1- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N, N-dimethylpyrrolidin-3-amine (13.0mg, 59%).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.59(d，1H，J＝2.2Hz)，8.01(d，1H，J＝2.3Hz)，4.90(m，1H)，4.40-3.60(m，3H)，2.90(m，1H)，2.39(s，3H)，2.37(s，3H)，2.30(m，1H)，2.00(m，1H)
Example 10
Synthesis of (R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine
(a) Synthesis of tert-butyl (R) - (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (100.0mg, 0.43mmol) and TEA (0.6mL, 4.30mmol) were dissolved in DCM (10.0mL) and tert-butyl (R) -pyrrolidin-3-ylcarbamate (94.0mg, 0.47mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain (R) - (1- (2, 7-dichloropyrido [2, 3-b) ] as a milky white solid]Pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester (169.0mg, 99%).
LC/MS ESI(+)：398(M+1)，400(M+3)
(b) Synthesis of (R) - (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester
Tert-butyl (R) - (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate (169.0mg, 0.42mmol) and hydrazine monohydrate (53.0mg, 1.05mmol) were dissolved in EtOH (5.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure to give the compound (R) - (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate as a yellow solid.
LC/MS ESI(+)：394(M+1)，396(M+3)
(c) Synthesis of tert-butyl (R) - (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamate
Unpurified (R) - (1- (7-chloro-2-hydrazinopyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate and trimethyl orthoformate (5.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure to obtain (R) - (1- (8-chloropyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：404(M+1)，406(M+3)
(d) Synthesis of (R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine
Unpurified (R) - (1- (8-chloropyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester was dissolved in DCM (3.0mL) to which TFA (1.0mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 90:10 to 50: 50). Collecting the product-containing fractions, and evaporating to obtain (R) -1- (8-chloropyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylpyrrolidin-3-amine (58.6 m)g, 45% and 3 steps).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.97(s，1H)，8.80(d，1H，J＝2.4Hz)，8.53(d，1H，J＝2.4Hz)，4.50-4.20(m，2H)，3.90-3.40(m，2H)，3.30(m，1H)，2.32(s，3H)，2.20-1.80(m，2H)
Example 11
Synthesis of (R) -1- (3-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylpyrrolidin-3-amine
The compound (R) -1- (3-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylpyrrolidin-3-amine was prepared as a cream white solid as in example 10 (18.0mg, 14%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.84(s，1H)，8.31(d，1H，J＝2.2Hz)，8.04(d，1H，J＝2.2Hz)，4.50-4.20(m，2H)，3.90-3.50(m，2H)，3.30(m，1H)，2.33(s，3H)，2.20-1.80(m，2H)
Example 12
Synthesis of 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine
5-bromopyridine-2, 3-diamine (5000.0mg, 2.66mmol) was added to diethyl oxalate (20.0mL), and the mixture was stirred at 120 ℃ for 12 hours, and then cooled to room temperature. Et was added thereto2O to form a solid. Then the solid formed was filtered and dried under reduced pressure to give 7-bromopyrido [2,3-b ] as a light brown solid]Pyrazine-2, 3-diols. Unpurified 7-bromopyrido [2,3-b ]]Pyrazine-2, 3-Diols and POCl3(20.0mL) of the mixture was stirred at 130 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid. The solid formed was filtered and then dried under reduced pressure to give 7-bromo-2, 3-dichloropyrido [2,3-b ] as a brown solid]Pyrazine (6500.0mg, 72%, 2 steps).
LC/MS ESI(+)：278(M+1)，280(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.28(d，1H，J＝2.4Hz)，8.99(d，1H，J＝2.4Hz)
(b) Synthesis of 7-bromo-2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
Reacting 7-bromo-2, 3-dichloropyrido [2,3-b ]]Pyrazine (1000.0mg, 3.59mmol) and TEA (5.0mL, 35.90mmol) were dissolved in DCM (20.0mL) and N-methylpiperazine (0.2mL, 2.01mmol) diluted with DCM (1.0mL) was added slowly thereto at-10 ℃. The reaction mixture was stirred for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 7-bromo-2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]Pyrazine (1000.0mg, 81%).
LC/MS ESI(+)：342(M+1)，344(M+3)
(c) Synthesis of 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 7-bromo-2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (413.0mg, 0.12mmol) and hydrazine monohydrate (150.0mg, 0.30mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 4 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid. The solid formed was then filtered and dried under reduced pressure to give 7-bromo-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]A pyrazine. Unpurified 7-bromo-2-hydrazino-3- (4-methylpiperazine-1)-yl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (10.0mL) was stirred at 80 ℃ for 2 hours and then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was then filtered and dried under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100:0-95: 5). Collecting the product-containing fractions, and evaporating to obtain 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazine (368.0mg, 29%, 3 steps).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，8.95(d，1H，J＝2.3Hz)，8.63(d，1H，J＝2.2Hz)，4.40(bs，4H)，2.53(m，4H)，2.24(s，3H)
Example 13
Synthesis of 4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile
Reacting 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (50.0mg, 0.14mmol), Zn (CN)2(17.0mg, 0.14mmol) and Pd (PPh)3)4(33.0mg, 0.01mmol) was dissolved in DMF (1.0mL) and allowed to react in a microwave reactor at 90 ℃ for 2 hours under 60W, then cooled to room temperature. The reaction mixture was then poured into water and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100:0-95: 5). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine-8-carbonitrile (6.0mg, 13%).
LC/MS ESI(+)：295(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，9.13(d，1H，J＝2.0Hz)，8.93(d，1H，J＝2.0Hz)，4.88(bs，2H)，4.14(bs，2H)，2.51(m，4H)，2.26(s，3H)
Example 14
Synthesis of 8-chloro-1-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.85mmol) and TEA (1180.0 μ L, 8.53mmol) were dissolved in DCM (8.5mL) and N-methylpiperazine (220.0 μ L, 0.94mmol) diluted with DCM (0.5mL) was added slowly at-20 ℃. The reaction mixture was stirred at-20 ℃ for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, Na-free2SO4Water drying, filtering, and then distilling under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). Collecting the product-containing fractions, and evaporating to obtain 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]Pyrazine (110.0mg, 43%).
LC/MS ESI(+)：298(M+1)，300(M+3)
1H-NMR(300MHz，DMSO-d6)：8.94(d，1H，J＝2.7Hz)，8.51(d，1H，J＝2.7Hz)，3.61(m，4H)，2.52(m，4H)，2.25(s，3H)。
(b) Synthesis of 7-chloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (100.0mg, 0.36mmol) and hydrazine monohydrate (46.0 μ L, 0.84mmol) were dissolved in EtOH (3.0mL), stirred at room temperature for 12 hours, and then evaporated under reduced pressure to give 7-chloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (50.0mg, 57%) as a yellow solid.
LC/MS ESI(+)：294(M+1)，296(M+3)
(c) Synthesis of 8-chloro-1-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 7-chloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine (50.0mg, 0.17mmol) and trimethyl orthoacetate (1.0mL) was stirred at 80 ℃ for 2 hours and then cooled to room temperature. Et was added thereto2O to form a solid, the solid formed was filtered and then dried under reduced pressure, and the residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 95: 5). Collecting the product-containing fractions, and evaporating to obtain milky white solid compound 8-chloro-1-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (30.0mg, 56%).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.56(d，1H，J＝2.4Hz)，8.42(d，1H，J＝2.4Hz)，4.36(m，4H)，3.02(s，3H)，2.49(m，4H)，2.25(s，3H)
Example 15
Synthesis of 8-chloro-1-methyl-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.85mmol) and TEA (1.2mL, 8.53mmol) were dissolved in DCM (8.5mL) and tert-butyl piperazine-1-carboxylate (174.7mg, 0.94mmol) diluted with DCM (0.5mL) was added slowly thereto at-20 ℃. The reaction mixture was stirred at-20 ℃ for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 4- (2, 7-bis) compound as yellow solidChloropyridino [2,3-b]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester (110.0mg, 64%).
LC/MS ESI(+)：384(M+1)，386(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.96(d，1H，J＝2.4Hz)，8.54(d，1H，J＝2.4Hz)，3.61(m，4H)，3.54(m，4H)，1.43(s，9H)
(b) Synthesis of tert-butyl 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
Tert-butyl 4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate (110.0mg, 0.29mmol) and hydrazine monohydrate (35.0 μ L, 0.72mmol) were dissolved in EtOH (2.0mL), stirred at room temperature for 12 hours, and evaporated under reduced pressure to give the yellow solid compound tert-butyl 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate (55.0mg, 51%).
LC/MS ESI(+)：380(M+1)，382(M+3)
(c) Synthesis of tert-butyl 4- (8-chloro-1-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate
Unpurified 4- (7-chloro-2-hydrazinopyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) piperazine-1-carboxylate (55.0mg, 0.15mmol) and trimethyl orthoacetate (0.7mL) was stirred at 80 ℃ for 2 hours, then cooled to room temperature. Et was added thereto2O and n-Hex (30.0mL) to form a solid, filtering the solid formed, and then drying under reduced pressure to give 4- (8-chloro-1-methylpyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) piperazine-1-carboxylic acid tert-butyl ester (35.0mg, 60%).
(d) Synthesis of 8-chloro-1-methyl-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified tert-butyl 4- (8-chloro-1-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate (35.0mg, 0.09mmol) was dissolved in DCM (2.0mL), to which TFA (0.5mL) was slowly added at room temperature, followed by stirring at room temperature for 2 hours. The reaction mixture was distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 80: 20). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-1-methyl-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a yellow solid (9.0mg, 35%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.55(m，1H)，8.42(m，1H)，4.34(m，4H)，3.04(s，3H)，2.87(m，4H)
Example 16
Synthesis of 8-bromo-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 4- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
Reacting 7-bromo-2, 3-dichloropyrido [2,3-b ]]Pyrazine (1000.0mg, 2.33mmol) and TEA (3.3mL, 23.30mmol) were dissolved in DCM (20.0mL) and tert-butyl piperazine-1-carboxylate (478.0mg, 2.56mmol) diluted with DCM (10.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give 4- (7-bromo-2-chloropyrido [2,3-b ] as a yellow solid]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：428(M+1)，430(M+3)
(b) Synthesis of tert-butyl 4- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
Unpurified 4- (7-bromo-2-chloropyrido [2, 3-b)]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester and hydrazine monohydrate (292.0mg, 5.83mmol) were dissolved in EtOH (50.0mL), stirred at room temperature for 12 hours,then, the mixture was distilled under reduced pressure. Et was added thereto2O to form a solid, filtering the solid formed, and then drying under reduced pressure to obtain 4- (7-bromo-2-hydrazinopyrido [2, 3-b) as a yellow solid compound]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：424(M+1)，426(M+3)
(c) Synthesis of tert-butyl 4- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate
Unpurified 4- (7-bromo-2-hydrazinopyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) piperazine-1-carboxylate and trimethyl orthoformate (10.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, and drying under reduced pressure to obtain 4- (8-bromopyrido [2, 3-e) as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：434(M+1)，436(M+3)
(d) Synthesis of 8-bromo-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 4- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) piperazine-1-carboxylic acid tert-butyl ester was dissolved in DCM (8.0mL) and TFA (2.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 8-bromo-4- (piperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (270.0mg, 35%, 3 steps).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，8.94(d，1H，J＝2.3Hz)，8.63(d，1H，J＝2.2Hz)，4.36(bs，4H)，2.89(m，4H)
Example 17
Synthesis of 7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 5, 6-dichloropyridine-2, 3-diamine
5-chloro-3-nitropyridin-2-amine (2000.0mg, 11.52mmol) and SnCl2(8740.0mg, 46.09mmol) was added to concentrated HCl (20.0mL) and stirred at 80-100 ℃ for 0.5 h. The reaction mixture was neutralized with saturated aqueous 1N NaOH (pH 7) and extracted with EtOAc (200.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc ═ 90: 10). The product-containing fractions were collected and evaporated to give 5, 6-dichloropyridine-2, 3-diamine as a milky white solid (1000.0mg, 49%).
1H-NMR(300MHz，DMSO-d6)；：6.80(s，1H)，6.04(s，2H)，5.11(s，2H)
(b) Synthesis of 2,3,6, 7-tetrachloropyrido [2,3-b ] pyrazine
5, 6-dichloropyridine-2, 3-diamine (1000.0mg, 5.62mmol) was added to diethyl oxalate (20.0mL), and the mixture was stirred at 120 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, drying under reduced pressure to obtain the brown solid compound 6, 7-dichloropyrido [2,3-b ]]Pyrazine-2, 3(1H,4H) -diones. Unpurified 6, 7-dichloropyrido [2,3-b ]]Pyrazine-2, 3(1H,4H) -diones and POCl3(20.0mL) of the mixture was stirred at 130 ℃ for 48 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid, and the formed solid was filtered and then dried under reduced pressure to obtain 2,3,6, 7-tetrachloropyrido [2,3-b ] as a brown solid compound]Pyrazine (1200.0mg, 72%, 2 steps).
1H-NMR(300MHz，DMSO-d6)；：9.09(s，1H)
(c) Synthesis of 2,6, 7-trichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3,6, 7-tetrachloropyrido [2,3-b ]]Pyrazine (190.0mg, 0.71mmol) and TEA (1.0mL, 7.10mmol) were dissolved in DCM (8.0mL), N-methylpiperazine (85.0mg, 0.85mmol) diluted with DCM (2.0mL) was added slowly thereto at-20 ℃ and stirred for 12 hours. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain yellow solid compound 2,6, 7-trichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A pyrazine.
LC/MS ESI(+)：332(M+1)，334(M+3)
(d) Synthesis of 7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 2,6, 7-trichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine and hydrazine monohydrate (89.0mg, 1.78mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 12 hours, and then distilled under reduced pressure to give 6, 7-dichloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid compound]A pyrazine. Unpurified 6, 7-dichloro-2-hydrazino-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (5.0mL) was stirred at 80 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazine (107.0mg, 45%, 3 steps).
LC/MS ESI(+)：338(M+1)，340(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.99(s，1H)，9.01(s，1H)，4.80-3.80(m，4H)，2.51(m，4H)，2.24(s，3H)
Example 18
Synthesis of 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
2,3, 7-trichloropyrido [2,3-b ] pyrazine (100.0mg, 0.43mmol) was dissolved in DCM (4.2mL), to which was added azetidin-3-yl (methyl) carbamic acid tert-butyl ester (88.0mg, 0.47mmol) and TEA (0.2mL, 1.28mmol) at 0 deg.C, followed by stirring for 1 hour. The solvent was removed from the reaction mixture under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (118.0mg, 72%) as a yellow solid.
LC/MS ESI(+)：384(M+1)，386(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.83(d，1H，J＝2.4Hz)，8.40(d，1H，J＝2.4Hz)，4.86(m，1H)，4.62(m，2H)，4.48(m，2H)，2.90(s，3H)，1.41(s，9H)
(b) Synthesis of tert-butyl (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
1- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (116.0mg, 0.30mmol) was dissolved in EtOH (4.2mL) and then hydrazine monohydrate (24.0. mu.L, 0.76mmol) was added thereto. The reaction mixture was stirred at room temperature for 1 hour. Et was added thereto2O to form a solid, filtering the solid formed, and then drying under reduced pressure to obtain the compound (1- (7-chloro-2-hydrazinopyrido [2, 3-b) as a yellow solid]Pyrazin-3-yl) azetidin-3-yl) (methyloxanecidYl) carbamic acid tert-butyl ester (114.0mg, 100%).
LC/MS ESI(+)：380(M+1)，382(M+3)
1H-NMR(300MHz，DMSO-d6)；：7.77(bs，1H)，7.25(bs，1H)，7.05(m，3H)，4.65(m，2H)，4.46(m，1H)，4.24(m，2H)，2.86(s，3H)，1.40(s，9H)
(c) Synthesis of tert-butyl (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Tert-butyl (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (119.0mg, 0.31mmol) was dissolved in trimethyl orthoformate (1.5mL) and then stirred at 75 ℃ for 1 hour. The reaction mixture was cooled to room temperature and then dried under reduced pressure to give tert-butyl (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (120.0mg, 98%) as a yellow solid.
LC/MS ESI(+)：390(M+1)，392(M+3)
(d) Synthesis of 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (120.0mg, 0.31mmol) was dissolved in DCM (0.6mL), and TFA (0.4mL) was added thereto. The reaction mixture was stirred at room temperature for 1 hour. The solvent was then removed from the reaction mixture under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated to yield 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine as a yellow solid (50.0mg, 57%).
LC/MS ESI(+)：290(M+1)，292(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.81(d，1H，J＝2.4Hz)，8.54(d，1H，J＝2.4Hz)，4.89(m，1H)，4.43(m，2H)，3.99(m，1H)，3.71(m，1H)，2.29(s，3H)
Example 19
Synthesis of (S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of (S) -4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.85mmol) and TEA (1180.0. mu.L, 8.53mmol) were dissolved in DCM (8.5mL) and tert-butyl (S) -2-methylpiperazine-1-carboxylate (187.8mg, 0.94mmol) diluted with DCM (0.5mL) was added slowly thereto at-20 ℃. The reaction mixture was stirred at-20 ℃ for 12 hours and poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 30: 70). The product-containing fractions were collected and evaporated to give (S) -4- (2, 7-dichloropyrido [2,3-b ] as a yellow solid]Pyrazin-3-yl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester (233.0mg, 67%).
LC/MS ESI(+)：398(M+1)，400(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.96(d，1H，J＝2.7Hz)，8.54(d，1H，J＝2.7Hz)，4.30(m，1H)，4.09(m，2H)，3.88(m，1H)，3.20(m，2H)，3.04(m，1H)，1.43(s，9H)，1.24(d，3H，J＝6.6Hz)
(b) Synthesis of (S) -4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester
Tert-butyl (S) -4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) -2-methylpiperazine-1-carboxylate (230.0mg, 0.58mmol) and hydrazine monohydrate (70.0. mu.L, 1.44mmol) were dissolved in EtOH (2.0 mL). The mixture was then stirred at room temperature under reduced pressure for 12 hours to give the compound (S) -tert-butyl 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -2-methylpiperazine-1-carboxylate (220.0mg, 97%) as a yellow solid.
LC/MS ESI(+)：394(M+1)，396(M+3)
(c) Synthesis of tert-butyl (S) -4- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -2-methylpiperazine-1-carboxylate
Unpurified (S) -4- (7-chloro-2-hydrazinopyrido [2, 3-b)]A mixture of pyrazin-3-yl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester (150.0mg, 0.38mmol) and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 2 hours, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure to obtain (S) -4- (8-chloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester (150.0mg, 81%).
(d) Synthesis of (S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified tert-butyl (S) -4- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -2-methylpiperazine-1-carboxylate (150.0mg, 0.37mmol) was dissolved in DCM (1.0 mL). TFA (0.2mL) was added slowly thereto at room temperature, followed by stirring at room temperature for 2 hours. The reaction mixture was distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 95: 5). The product-containing fractions were collected and evaporated to yield the compound (S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a yellow solid (70.0mg, 63%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.61(d，1H，J＝2.4Hz)，8.04(d，1H，J＝2.4Hz)，6.24(m，1H)，5.21(m，1H)，3.02(m，5H)，1.21(d，3H，J＝6Hz)
Example 20
Synthesis of (S) -8-chloro-4- (3, 4-dimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2, 3-e) obtained in example 19(d)][1,2,4]Triazolo [4,3-a]Pyrazine (50.0mg, 0.16mmol) and 37% formaldehyde (73.5 μ L, 0.99mmol) were dissolved in MeOH (0.5 mL). To which NaBH was slowly added4(37.0mg, 0.99mmol), followed by stirring at room temperature for 12 hours. Brine was poured into the reaction mixture, extracted with DCM (30.0mL), and extracted with anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 95:5) to give (S) -8-chloro-4- (3, 4-dimethylpiperazin-1-yl) pyrido [2,3-e ] as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazine (70.0mg, 48%).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.62(d，1H，J＝2.4Hz)，8.03(d，1H，J＝2.1Hz)，6.06(m，1H)，5.12(m，1H)，3.56(m，2H)，2.99(m，1H)，2.37(m，5H)，1.22(d，3H，J＝6.3Hz)
Example 21
Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile
Mixing 7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (10.0mg, 0.03mmol), Zn (CN)2(4.0mg, 0.03mmol) and Pd (PPh)3)4(3.0mg, 0.003mmol) was dissolved in DMF (1.0 mL). The mixture was reacted in a microwave reactor at 60W at 100 ℃ for 1 hour, and then cooled to room temperature. The reaction mixture was poured into water and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (D)CM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine-7-carbonitrile (2.0mg, 20%).
LC/MS ESI(+)：329(M+1)，331(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.04(s，1H)，9.07(s，1H)，4.80(m，2H)，4.09(m，2H)，2.54(m，4H)，2.25(s，3H)
Example 22
Synthesis of 8-chloro-4- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 3, 5-dimethylpiperazine-1-carboxylate
2, 6-dimethylpiperazine (200.0mg, 1.75mmol) and TEA (0.6mL, 4.37mmol) were dissolved in DCM (6.0mL) and added slowly (Boc) thereto at 0 deg.C2O (458.7mg, 2.10 mmol). The reaction mixture was stirred at room temperature for 12 hours, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 95: 5). The product-containing fractions were collected and evaporated to give 3, 5-dimethylpiperazine-1-carboxylic acid tert-butyl ester as a yellow liquid (210.0mg, 56%).
1H-NMR(300MHz，CDCl3)；：3.95(m，2H)，2.79(m，2H)，2.33(m，2H)，1.46(s，9H)，1.07(d，6H，J＝6.3Hz)
(b) Synthesis of tert-butyl 3,4, 5-trimethylpiperazine-1-carboxylate
Tert-butyl 3, 5-dimethylpiperazine-1-carboxylate (200.0mg, 0.93mmol) and 37% formaldehyde (440.0. mu.L, 5.56mmol) were dissolved in MeOH (5.0mL), to which NaBH was slowly added4(172.6mg, 5.56mmol) and stirred at room temperature for 12 hours. Brine was poured into the reaction mixture and extracted with DCM (30.0 mL). With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. Will remainThe product was purified by silica gel column chromatography (DCM: MeOH ═ 95:5) to give compound 3,4, 5-trimethylpiperazine-1-carboxylic acid tert-butyl ester (57.0mg, 27%) as a yellow liquid.
(c) Synthesis of 1,2, 6-trimethylpiperazine
Tert-butyl 3,4, 5-trimethylpiperazine-1-carboxylate (57.0mg, 0.25mmol) was dissolved in DCM (1.0mL) and TFA (0.2mL) was added slowly thereto at room temperature. The reaction mixture was stirred at room temperature for 2 hours, then distilled under reduced pressure, and purified by amine silica gel column chromatography (DCM: MeOH ═ 95: 5). The product-containing fractions were collected and evaporated to yield the compound 1,2, 6-trimethylpiperazine (32.0mg, 100%) as a yellow liquid.
1H-NMR(300MHz，CDCl3)；：2.88(m，2H)，2.58(m，2H)，2.28(s，3H)，2.14(m，2H)，1.07(d，6H，J＝6Hz)
(d) Synthesis of 2, 7-dichloro-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (58.4mg, 0.25mmol) and TEA (347.0 μ L, 2.49mmol) were dissolved in DCM (1.0mL) and 1,2, 6-trimethylpiperazine (32.0mg, 0.25mmol) in DCM (0.5mL) was added slowly thereto at-20 ℃. The reaction mixture was stirred at-20 ℃ for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and evaporating under reduced pressure to obtain 2, 7-dichloro-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b ] as yellow solid]Pyrazine (23.0mg, 28%).
LC/MS ESI(+)：326(M+1)，328(M+3)
(e) Synthesis of 7-chloro-2-hydrazino-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2, 7-dichloro-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (23.0mg, 0.07mmol) and hydrazine monohydrate (8.0 μ L, 0.18mmol) were dissolved in EtOH (0.3mL), stirred at room temperature for 12 hours, and then evaporated under reduced pressure to give 7-chloro-2-hydrazino-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b ] pyrazine as a yellow solid (23.0mg, 100%).
LC/MS ESI(+)：322(M+1)，324(M+3)
(f) Synthesis of 8-chloro-4- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Unpurified 7-chloro-2-hydrazino-3- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine (23.0mg, 0.07mmol) and trimethyl orthoformate (2.0mL) was stirred at 80 deg.C for 2 hours, then cooled to room temperature. Adding Et thereto2O to form a solid, filtering the solid formed and then drying under reduced pressure to obtain 8-chloro-4- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (5.0mg, 21%).
LC/MS ESI(+)：332(M+1)，334(M+3)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.62(d，1H，J＝2.4Hz)，8.04(d，1H，J＝2.4Hz)，6.17(m，1H)，5.18(m，1H)，3.28(m，1H)，2.93(m，1H)，2.39(m，2H)，2.34(s，3H)1.26(s，6H)
Example 23
Synthesis of 8-chloro-7-ethoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (20.0mg, 0.06mmol) and NaOEt (5.0mg, 0.07mmol) were dissolved in EtOH (1.0mL), reacted in a microwave reactor at 60W at 90 ℃ for 2 hours, and then cooled to room temperature. The reaction mixture was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-7-ethoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream white solid (3.0mg, 14%).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.91(s，1H)，8.84(s，1H)，4.44(q，2H，J＝7.0Hz)，4.40(m，4H)，2.50(m，4H)，2.24(s，3H)，2.24(t，3H，J＝7.0Hz)
Example 24
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
Reacting 7-bromo-2, 3-dichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.72mmol) and TEA (1.0mL, 7.20mmol) were dissolved in DCM (8.0mL) and tert-butyl azetidin-3-yl (methyl) carbamate (147.0mg, 0.79mmol) diluted with DCM (2.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and then poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give the brown solid compound (1- (7-bromo-2-chloropyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：428(M+1)，430(M+3)
(b) Synthesis of tert-butyl (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
Unpurified (1- (7-bromo-2-chloropyrido [2, 3-b))]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (224.0mg, 0.61mmol) and hydrazine monohydrate (72.0mg, 3.78mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 12 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid, filter shapeThe resulting solid was then dried under reduced pressure to give the compound (1- (7-bromo-2-hydrazinopyrido [2, 3-b) ] as a yellow solid]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：424(M+1)，426(M+3)
(c) Synthesis of tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Unpurified (1- (7-bromo-2-hydrazinopyrido [2, 3-b))]A mixture of t-butyl pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate and trimethyl orthoformate (10.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, and drying under reduced pressure to obtain a milky white solid compound (1- (8-bromopyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (137.0mg, 45%, 3 steps).
LC/MS ESI(+)：434(M+1)，436(M+3)
(d) Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
1- (8-bromopyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (87.0mg, 0.20mmol) was dissolved in DCM (4.0mL) to which TFA (1.0mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (16.0mg, 23%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.90(d，1H，J＝2.1Hz)，8.59(d，1H，J＝2.1Hz)，4.89(m，1H)，4.42(m，2H)，4.00(m，1H)，3.73(m，1H)，2.29(s，3H)
Example 25
Synthesis of 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile
1- (8-bromopyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (50.0mg, 0.12mmol), Zn (CN)2(14.0mg, 0.12mmol) and Pd (PPh)3)4(13.0mg, 0.01mmol) was dissolved in DMF (1.0mL) and allowed to react in a microwave reactor at 60W at 90 ℃ for 2 hours, then cooled to room temperature. The reaction mixture was then purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to give (1- (8-cyanopyrido [2, 3-e) as a cream-white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester. Reacting (1- (8-cyanopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester was dissolved in DCM (4.0mL) to which TFA (1.0mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). Collecting the product-containing fractions, evaporating to obtain a milky white solid compound 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine-8-carbonitrile (10.0mg, 22%).
LC/MS ESI(+)：281(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.93(s，1H)，9.06(d，1H，J＝1.9Hz)，8.88(d，1H，J＝1.9Hz)，4.95(m，1H)，4.49(m，2H)，4.04(m，1H)，3.73(m，1H)，2.30(s，3H)
Example 26
Synthesis of 4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile
Reacting 8-bromo-4- (piperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (50.0mg, 0.15mmol), Zn (CN)2(18.0mg, 0.15mmol) and Pd (PPh)3)4(3.0mg, 0.02mmol) was dissolved in DMF (1.0mL) and allowed to react in a microwave reactor at 60W at 90 ℃ for 1 hour, then cooled to room temperature. The reaction mixture was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). Collecting the product-containing fractions, evaporating to obtain a milky white solid compound 4- (piperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine-8-carbonitrile (4.4mg, 10%).
LC/MS ESI(+)：281(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.98(s，1H)，9.09(d，1H，J＝2.0Hz)，8.90(d，1H，J＝2.1Hz)，4.80(m，2H)，4.05(m，2H)，2.90(m，4H)
Example 27
Synthesis of 1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of methyl (1- (2,6, 7-trichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester
2,3,6, 7-tetrachloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.74mmol) and TEA (1.0mL, 7.40mmol) were dissolved in DCM (8.0mL) and tert-butyl azetidin-3-yl (methyl) carbamate (162.0mg, 0.9 mL) diluted with DCM (2.0mL) was added slowly thereto at 0 deg.C4 mmol). The reaction mixture was stirred at room temperature for 12 hours and poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give methyl (1- (2,6, 7-trichloropyrido [2, 3-b) as a brown solid]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：418(M+1)，420(M+3)
(b) Synthesis of tert-butyl (1- (6, 7-dichloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
The unpurified methyl (1- (2,6, 7-trichloropyrido [2, 3-b))]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester and hydrazine monohydrate (37.0mg, 1.85mmol) were dissolved in EtOH (10.0mL), then stirred at room temperature for 12 hours, then distilled under reduced pressure. Et was added thereto2O to form a solid, filtering the solid formed, and then drying under reduced pressure to obtain the compound (1- (6, 7-dichloro-2-hydrazinopyrido [2, 3-b) as a yellow solid]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：414(M+1)，416(M+3)
(c) Synthesis of tert-butyl (1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Unpurified (1- (6, 7-dichloro-2-hydrazinopyrido [2, 3-b))]A mixture of t-butyl pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate and trimethyl orthoformate (20.0mL) was stirred at 70 ℃ for 1 hour, then cooled to room temperature. Et was added slowly thereto2O, filtering the solid formed, then drying under reduced pressure to obtain the compound (1- (7, 8-dichloropyrido [2,3-e ]) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (173.0mg, 55%, 3 steps).
LC/MS ESI(+)：424(M++1)，426(M+3)
(d) Synthesis of 1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
1- (7, 8-dichloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (123.0mg, 0.29mmol) was dissolved in DCM (4.0mL) to which TFA (1.0mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (7, 8-dichloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (19.0mg, 20%).
LC/MS ESI(+)：324(M+1)，326(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.98(s，1H)，4.94(m，1H)，4.46(m，2H)，4.00(m，1H)，3.73(m，1H)，2.30(s，3H)
Example 28
Synthesis of 8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile
1- (7, 8-dichloropyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (50.0mg, 0.15mmol), Zn (CN)2(18.0mg, 0.15mmol) and Pd (PPh)3)4(18.0mg, 0.02mmol) was dissolved in DMF (1.0mL) and allowed to react in a microwave reactor at 60W at 90 ℃ for 1 hour, then cooled to room temperature. The reaction mixture was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). MiningThe product-containing fractions were collected and evaporated to give (1- (8-chloro-7-cyanopyrido [2, 3-e) as a cream-white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester. Unpurified (1- (8-chloro-7-cyanopyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester was dissolved in DCM (2.0mL) to which TFA (0.5mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, and then distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). Collecting the product-containing fractions, evaporating to obtain a milky white solid compound 8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine-7-carbonitrile (20.0mg, 41%).
LC/MS ESI(+)：315(M+1)，317(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.98(s，1H)，9.03(s，1H)，4.93(m，1H)，4.48(m，2H)，4.00(m，1H)，3.73(m，1H)，2.30(s，3H)
Example 29
Synthesis of 8-chloro-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ] pyrazine (100.0mg, 0.43mmol) and octahydropyrrolo [1,2-a ] pyrazine (60.0mg, 0.47mmol) were reacted in the same manner as in example 18(a) to give 2, 7-dichloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-b ] pyrazine (83.0mg, 60%) as a yellow solid compound.
LC/MS ESI(+)：324(M+1)，326(M+3)
1H-NMR(300MHz，CDCl3)；：8.86(d，1H，J＝2.7Hz)，8.17(d，1H，J＝2.4Hz)，4.45(m，2H)，3.32(m，1H)，3.19(m，2H)，2.97(m，1H)，2.49(m，1H)，2.26(m，2H)，1.91(m，2H)，1.81(m，1H)，1.51(m，1H)
(b) Synthesis of 7-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -2-hydrazinopyrido [2,3-b ] pyrazine
2, 7-dichloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-b ] pyrazine (80.0mg, 0.25mmol) and hydrazine monohydrate (19.0 μ L, 0.62mmol) were reacted in the same manner as in example 18(b) to give 7-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -2-hydrazinopyrido [2,3-b ] pyrazine (50.0mg, 63%) as an orange solid.
LC/MS ESI(+)：320(M+1)，322(M+3)
1H-NMR(300MHz，CDCl3)；：8.60(d，1H，J＝3.0Hz)，8.01(d，1H，J＝3.0Hz)，6.49(bs，1H)，4.20(d，2H，J＝3.0Hz)，3.94(m，2H)，3.17(m，3H)，2.87(m，1H)，2.46(m，1H)，2.22(m，2H)，1.89(m，2H)，1.78(m，1H)，1.50(m，1H)
(c) Synthesis of 8-chloro-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
7-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -2-hydrazinopyrido [2,3-b ] pyrazine (45.0mg, 0.14mmol) and trimethyl orthoformate (1.0mL) were reacted at 90 ℃ for 5 hours in the same manner as in example 18(c) to give 8-chloro-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (37.0mg, 79%) as an orange solid.
LC/MS ESI(+)：330(M+1)，332(M+3)
1H-NMR(300MHz，CDCl3)；：9.16(s，1H)，8.61(d，1H，J＝2.1Hz)，8.04(d，1H，J＝2.4Hz)，6.39(m，1H)，5.50(m，1H)，3.55(m，0.5H)，3.20(m，3H)，2.90(m，0.5H)，2.42(m，1H)，2.20(m，2H)，1.88(m，3H)，1.52(m，1H)
Example 30
Synthesis of (R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-amine
(a) Synthesis of tert-butyl (R) - (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) carbamate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (200.0mg, 0.85mmol) and TEA (1.2mL, 8.50mmol) were dissolved in DCM (10.0mL) and (R) -pyrrolidin-3-ylcarbamic acid tert-butyl ester (175.0mg, 0.94mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 100:0-98: 2). Collecting the product-containing fractions, and evaporating to obtain (R) - (1- (2, 7-dichloropyrido [2, 3-b) ] as yellow solid]Pyrazin-3-yl) pyrrolidin-3-yl) carbamic acid tert-butyl ester (284.0mg, 86%).
LC/MS ESI(+)：384(M+1)，386(M+3)
(b) Synthesis of (R) - (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) carbamic acid tert-butyl ester
Tert-butyl (R) - (1- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) carbamate (284.0mg, 0.74mmol) and hydrazine monohydrate (92.0mg, 1.85mmol) were dissolved in EtOH (10.0mL), stirred at room temperature for 2 hours, and then distilled under reduced pressure to give the compound (R) - (1- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) carbamic acid tert-butyl ester as a yellow solid.
LC/MS ESI(+)：380(M+1)，382(M+3)
(c) Synthesis of tert-butyl (R) - (1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-yl) carbamate
Unpurified (R) - (1- (7-chloro-2-hydrazinopyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) pyrrolidin-3-yl) carbamate and trimethyl orthoformate (5.0mL) was stirred at 70 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure to obtain the compound (R) - (1- (8-chloropyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-yl) carbamic acid tert-butyl ester (272.0mg, 94%, 2 steps).
LC/MS ESI(+)：390(M+1)，392(M+3)
(d) Synthesis of (R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-amine
Reacting (R) - (1- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-yl) carbamic acid tert-butyl ester (272.0mg, 0.70mmol) was dissolved in DCM (20.0mL) and TFA (4.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and extracted with DCM (30.0 mL). The organic layer was washed with brine, Na-free2SO4Water drying, filtering, and then distilling under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 90:10 to 50: 50). Collecting the product-containing fractions, and evaporating to obtain (R) -1- (8-chloropyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) pyrrolidin-3-amine (79.0mg, 39%).
LC/MS ESI(+)：290(M+1)，292(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.99(s，1H)，8.81(d，1H，J＝2.4Hz)，8.53(d，1H，J＝2.4Hz)，4.6-3.4(m，5H)，2.4-1.6(m，2H)
Example 31
Synthesis of 9-chloro-2-methyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of 3-bromo-5-chloropyridin-2-amine
5-Chloropyridin-2-amine (5000.0mg, 38.90mmol) was dissolved in CHCl3(78.0mL), to which Br was then added2(2.0mL, 38.90 mmol). The reaction mixture was stirred at room temperature for 1 hour, and the solvent was removed therefrom under reduced pressure. The residue was dissolved in EtOAc and then saturated NaHCO3Aqueous solution and brine. With anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 9). The product-containing fractions were collected and evaporated to give 3-bromo-5-chloropyridin-2-amine (7500.0mg, 93%) as a yellow solid.
LC/MS ESI(+)：207(M+1)，209(M+3)
1H-NMR(300MHz，CDCl3)；：7.98(d，1H，J＝2.1Hz)，7.66(d，1H，J＝2.1Hz)，4.93(bs，2H)
(b) Synthesis of 2-amino-5-chloronicotinonitrile
3-bromo-5-chloropyridin-2-amine (2700.0mg, 13.00mmol) was dissolved in NMP (60.0mL), to which was then added Zn (CN)2(2300.0mg, 19.50mmol) and Pd (PPh)3)4(1500.0mg, 1.30 mmol). The reaction mixture was stirred at 110 ℃ for 5 hours and then cooled to room temperature. Water and EtOAc were added to the reaction mixture, stirred for 10 minutes, then filtered through celite. The filtrate was extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 9). The product-containing fractions were collected and evaporated to give 2-amino-5-chloronicotinonitrile (1900.0mg, 100%) as a white solid.
LC/MS ESI(+)：154(M+1)，156(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.22(d，1H，J＝2.7Hz)，8.07(d，1H，J＝2.7Hz)，7.14(s，2H)
(c) Synthesis of 9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol
2-amino-5-chloronicotinonitrile (300.0mg, 1.95mmol) was dissolved in MEK (2.0mL) and NaHCO was added thereto3(492.0mg, 5.86mmol) and chloroethyl formate (3.0 mL). The reaction mixture was refluxed, then cooled to room temperature, filtered, and then dried under reduced pressure to give the atypical yellow compound ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate. Unpurified ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate was dissolved in diphenyl ether (2.0mL) and acethydrazide (144.0mg, 1.95mmol) was added thereto. The reaction mixture was stirred at 180 ℃ for 30 minutes, then cooled to room temperature, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). Collecting the product-containing fractions, and evaporating to obtain 9-chloro-2-methylpyrido [3,2-e ] as a yellow solid][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (91.0mg, 20%, 2 steps).
LC/MS ESI(+)：236(M+1)，238(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.9(s，1H)，8.73(d，1H，J＝2.4Hz)，8.58(d，1H，J＝2.4Hz)
(d) Synthesis of 5, 9-dichloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
Reacting 9-chloro-2-methylpyrido [3,2-e ]][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (80.0mg, 0.34mmol) dissolved in POCl3(1.5mL), to which was added DIPEA (120.0. mu.L, 0.68 mmol). The reaction mixture was refluxed for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3Aqueous solution was neutralized and then extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to give a white colorSolid compound 5, 9-dichloro-2-methylpyrido [3,2-e][1,2,4]Triazolo [1,5-c]Pyrimidine (30.0mg, 35%).
LC/MS ESI(+)：254(M+1)，256(M+3)
(e) Synthesis of 9-chloro-2-methyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (28.0mg, 0.11mmol) was dissolved in DMF (1.1mL), to which was added N-methylpiperazine (24.0. mu.L, 0.22 mmol). The reaction mixture was stirred at room temperature for 1 hour, and the residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give the compound 9-chloro-2-methyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a cream white solid (18.0mg, 51%).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.84(d，1H，J＝2.4Hz)，8.63(d，1H，J＝2.4Hz)，4.12(m，4H)，2.56(s，3H)，2.54(m，4H)，2.24(s，3H)
Example 32
Synthesis of 9-chloro-2-methyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of tert-butyl 4- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) piperazine-1-carboxylate
5, 9-dichloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine from example 31(d) (20.0mg, 0.08mmol) was dissolved in DMF (1.0mL), to which was added piperazine-1-carboxylic acid tert-butyl ester (29.0mg, 0.16 mmol). The reaction mixture was stirred at room temperature for 1 hour and purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to give tert-butyl 4- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) piperazine-1-carboxylate (32.0mg, 100%) as a cream-white solid.
LC/MS ESI(+)：404(M+1)，406(M+3)
(b) Synthesis of 9-chloro-2-methyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
Tert-butyl-4- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) piperazine-1-carboxylate (30.0mg, 0.07mmol) was dissolved in DCM (0.6mL), to which was added TFA (0.4 mL). The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give the compound 9-chloro-2-methyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a cream-white solid (22.5mg, 100%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：8.79(d，1H，J＝2.7Hz)，8.58(d，1H，J＝2.7Hz)，4.26(m，4H)，3.11(m，4H)，2.64(s，3H)
Example 33
Synthesis of 1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate
5, 9-dichloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine from example 31(d) (12.0mg, 0.05mmol) was dissolved in DMF (1.0mL), to which was added tert-butyl azetidin-3-yl (methyl) carbamate (17.6mg, 0.09 mmol). The reaction mixture was stirred at room temperature for 1 hour, from which the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give tert-butyl (1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate as a cream solid (19.0mg, 100%).
LC/MS ESI(+)：404(M+1)，406(M+3)
(b) Synthesis of 1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine
Tert-butyl (1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate (18.0mg, 0.04mmol) was dissolved in DCM (0.6mL), to which was added TFA (0.4 mL). The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give the compound 1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine as a cream white solid (7.0mg, 55%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：8.73(d，1H，J＝2.7Hz)，8.51(d，1H，J＝2.7Hz)，4.84(m，2H)，4.38(m，2H)，3.80(m，1H)，2.60(s，3H)，2.49(s，3H)
Example 34
Synthesis of 9-chloro-2-cyclopropyl-N, N-diethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-amine
(a) Synthesis of 9-chloro-2-cyclopropylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol
Ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate was synthesized from 2-amino-5-chloronicotinonitrile (500.0mg, 3.26mmol) and reacted with cyclopropanecarbohydrazide (326.0mg, 3.26mmol) in the same manner as in example 31(c) to give 9-chloro-2-cyclopropylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol as a brown solid (37.0mg, 4%, 2 steps).
LC/MS ESI(+)：262(M+1)，264(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.86(bs，1H)，8.71(d，1H，J＝2.7Hz)，8.53(d，1H，J＝2.7Hz)，2.23(m，1H)，1.09(m，2H)，0.99(m，2H)
(b) Synthesis of 9-chloro-2-cyclopropyl-N, N-diethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-amine
Reacting 9-chloro-2-cyclopropyl pyrido [3,2-e ]][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (35.0mg, 0.13mmol) in POCl3(1.5mL), to which TEA (37.0. mu.L, 0.27mmol) was added. The reaction mixture was refluxed for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3The aqueous solution was neutralized and extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and evaporated to give the yellow solid compound 9-chloro-2-cyclopropyl-N, N-diethylpyrido [3,2-e][1,2,4]Triazolo [1,5-c]Pyrimidin-5-amine (32.0mg, 74%).
LC/MS ESI(+)：317(M+1)，319(M+3)
1H-NMR(300MHz，CDCl3)；：8.71(d，1H，J＝2.7Hz)，8.51(d，1H，J＝2.7Hz)，4.04(q，4H，J＝6.9Hz)，2.23(m，1H)，1.39(t，6H，J＝6.9Hz)，1.13(m，4H)
Example 35
Synthesis of 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of 9-chloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5 (6H) -one
2-amino-5-chloronicotinonitrile (500.0mg, 3.26mmol) was dissolved in MEK (3.5mL) and the solution was stirredWherein NaHCO is added3(820.4mg, 9.77mmol) and chloroethyl formate (5.0 mL). The reaction mixture was refluxed for 24 hours, then cooled to room temperature, filtered, and then dried under reduced pressure to give the atypical yellow compound ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate. Unpurified ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate was dissolved in diphenyl ether (3.5mL), and then formylhydrazine (195.5mg, 3.26mmol) was added thereto. The reaction mixture was stirred at 180 ℃ for 1 hour, then cooled to room temperature and purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 1). Collecting the product-containing fractions, and evaporating to obtain 9-chloropyrido [3,2-e ] as a yellow solid][1,2,4]Triazolo [1,5-c]Pyrimidin-5 (6H) -one (63.0mg, 9%).
LC/MS ESI(+)：221(M+1)，223(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.95(s，1H)，8.74(d，1H，J＝2.1Hz)，8.63(d，1H，J＝2.4Hz)，8.59(s，1H)。
(b) Synthesis of 5, 9-dichloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
Reacting 9-chloropyrido [3,2-e ]][1,2,4]Triazolo [1,5-c]Pyrimidin-5 (6H) -one (63.0mg, 0.29mmol) in POCl3(2.0mL) DIPEA (150.0. mu.L, 0.86mmol) was added. The reaction mixture was refluxed for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3The aqueous solution was neutralized and extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 1). Collecting the product-containing fractions, and evaporating to obtain 5, 9-dichloropyrido [3,2-e ] as a white solid][1,2,4]Triazolo [1,5-c]Pyrimidine (50.4mg, 72%).
LC/MS ESI(+)：240(M+1)，241(M+3)
(c) Synthesis of 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (15.0mg, 0.06mmol) was dissolved in DMF (0.5mL), to which was added N-methylpiperazine (10.3. mu.L, 0.09mmol) and TEA (25.0. mu.L, 0.19 mmol). The reaction mixture was stirred at room temperature for 1 hour, then purified by silica gel column chromatography (MeOH: DCM ═ 1: 19). The product-containing fractions were collected and evaporated to give the compound 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a cream-white solid (6.0mg, 33%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：8.82(d，1H，J＝3.0Hz)，8.63(d，1H，J＝2.7Hz)，8.37(s，1H)，4.33(m，4H)，2.65(m，4H)，2.38(s，3H)。
Example 36
Synthesis of 1- (9-chloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine
5, 9-dichloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine from example 35(b) (15.0mg, 0.06mmol) was dissolved in DMF (0.5mL), and TEA (25.0. mu.L, 0.19mmol) was added t-butyl azetidin-3-yl (methyl) carbamate (17.3mg, 0.09 mmol). The reaction mixture was stirred at room temperature for 1 hour, distilled under reduced pressure, and then dissolved in DCM (1.0mL) without purification, and then TFA (0.2mL) was added thereto. The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 19). The product-containing fractions were collected and evaporated to give the compound 1- (9-chloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine as a cream-white solid (2.6mg, 15%).
LC/MS ESI(+)：290(M+1)，292(M+3)
1H-NMR(300MHz，CDCl3+MeOH-d4)；：8.70(d，1H，J＝2.7Hz)，8.53(d，1H，J＝2.7Hz)，8.28(d，1H，J＝2.7Hz)，4.84(m，2H)，4.45(m，2H)，3.82(m，1H)，2.46(s，3H)。
Example 37
Synthesis of 9-chloro-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine from example 35(b) (15.0mg, 0.06mmol) was dissolved in DMF (0.5mL), and then piperazine-1-carboxylic acid tert-butyl ester (16.7mg, 0.09mmol) and TEA (25.0. mu.L, 0.19mmol) were added thereto. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure, then dissolved in DCM (1.0mL) without purification, to which TFA (0.2mL) was added. The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 19). The product-containing fractions were collected and evaporated to yield the compound 9-chloro-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a white solid (1.7mg, 10%).
LC/MS ESI(+)：290(M+1)，292(M+3)
1H-NMR(300MHz，CDCl3+MeOH-d4)；：8.82(d，1H，J＝2.4Hz)，8.68(d，1H，J＝2.7Hz)，8.38(s，1H)，4.51(m，4H)，3.39(m，4H)。
Example 38
Synthesis of 9-chloro-2-cyclopropyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of 5, 9-dichloro-2-cyclopropylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
9-chloro-2-cyclopropylpyrido [3,2-e ] was reacted in the same manner as in example 31(d)][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (125.0mg, 0.48mmol) with POCl3(2.5mL) and DIPEA (0.2mL, 0.96mmol) to give the compound 5, 9-dichloro-2-cyclopropylpyrido [3,2-e ] as a milky white solid][1,2,4]Triazolo [ 1],5-c]Pyrimidine (90.0mg, 67%).
LC/MS ESI(+)：280(M+1)，282(M+3)
1H-NMR(300MHz，CDCl3)；：9.01(d，1H，J＝2.7Hz)，8.76(d，1H，J＝2.7Hz)，2.34(m，1H)，1.22(m，4H)
(b) Synthesis of 9-chloro-2-cyclopropyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloro-2-cyclopropylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (25.0mg, 0.09mmol) and N-methylpiperazine (20.0. mu.L, 0.18mmol) were reacted in the same manner as in example 31(e) to give 9-chloro-2-cyclopropyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a milky white solid (22.0mg, 72%).
LC/MS ESI(+)：344(M+1)，346(M+3)
1H-NMR(300MHz，CDCl3)；：8.77(d，1H，J＝2.7Hz)，8.56(d，1H，J＝2.7Hz)，4.31(t，4H，J＝5.1Hz)，2.64(t，4H，J＝5.1Hz)，2.38(s，3H)，2.25(m，1H)，1.15(m，4H)
Example 39
Synthesis of 9-chloro-2-cyclopropyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloro-2-cyclopropylpyrido [3,2-e ] obtained in the manner of example 38(a)][1,2,4]Triazolo [1,5-c]Pyrimidine (30.0mg, 0.11mmol) was dissolved in DCM (1.0mL), to which was added piperazine-1-carboxylic acid tert-butyl ester (40.0mg, 0.21 mmol). The reaction mixture was stirred for 1 hour, and then TFA (0.8mL) was added and allowed to react for another 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Et was added thereto 2O and n-Hex, filtering the resulting solid to obtain a milky white solid compound 9-chloro-2-cyclopropyl-5- (piperazine)Oxazin-1-yl) pyrido [3,2-e][1,2,4]Triazolo [1,5-c]Pyrimidine (31.0mg, 89%).
LC/MS ESI(+)：330(M+1)，332(M+3)
1H-NMR(300MHz，CDCl3)；：8.77(d，1H，J＝2.7Hz)，8.56(d，1H，J＝2.7Hz)，4.25(t，4H，J＝5.1Hz)，3.10(t，4H，J＝5.1Hz)，2.25(m，1H)，1.15(m，4H)
Example 40
Synthesis of 9-chloro-2- (methoxymethyl) -5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of 9-chloro-2- (methoxymethyl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol
2-amino-5-chloronicotinonitrile (500.0mg, 3.26mmol) was dissolved in MEK (4.0mL) and K2CO3(1350.0mg, 9.78mmol), to which MgSO was added4(250.0mg, 1.25mmol) and chloroethyl formate (6.0 mL). The reaction mixture was refluxed for 24 hours, then cooled to room temperature, filtered, and then dried under reduced pressure to give the atypical yellow compound ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate. Unpurified ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate was dissolved in diphenyl ether (2.0mL) and 2-methoxyacetohydrazide (271.0mg, 2.61mmol) was added. The reaction mixture was stirred at 150 ℃ for 3 hours, then cooled to room temperature, and then purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and evaporated to give the compound 9-chloro-2- (methoxymethyl) pyrido [3,2-e as a yellow solid][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (150.0mg, 17%, 2 steps).
LC/MS ESI(+)：266(M+1)，268(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.99(s，1H)，8.74(d，1H，J＝2.4Hz)，8.64(d，1H，J＝2.7Hz)，4.63(s，2H)，3.38(s，3H)
(b) Synthesis of 5, 9-dichloro-2- (methoxymethyl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
9-chloro-2- (methoxymethyl) pyrido [3,2-e ] was synthesized in the same manner as in example 31(d)][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (146.0mg, 0.51mmol) with POCl3(1.5mL) and DIPEA (0.2mL, 1.03mmol) to give the compound 5, 9-dichloro-2- (methoxymethyl) pyrido [3,2-e as a milky white solid][1,2,4]Triazolo [1,5-c]Pyrimidine (89.0mg, 61%).
LC/MS ESI(+)：284(M+1)，286(M+3)
1H-NMR(300MHz，CDCl3)；：9.06(d，1H，J＝2.4Hz)，8.87(d，1H，J＝2.7Hz)，4.85(s，2H)，3.59(s，3H)
(c) Synthesis of 9-chloro-2- (methoxymethyl) -5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloro-2- (methoxymethyl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (25.0mg, 0.09mmol) and N-methylpiperazine (20.0. mu.L, 0.18mmol) were reacted in DCM (1.0mL) in the same manner as in example 31(e) to give 9-chloro-2- (methoxymethyl) -5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a milky white solid (18.0mg, 58%).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，CDCl3)；：8.81(d，1H，J＝2.7Hz)，8.67(d，1H，J＝2.7Hz)，4.76(s，2H)，4.33(t，4H，J＝4.8Hz)，3.57(s，3H)，2.64(t，4H，J＝4.8Hz)，2.38(s，3H)
EXAMPLE 41
Synthesis of 9-chloro-2-ethyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
(a) Synthesis of 9-chloro-2-ethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol
Ethyl (5-chloro-3-cyanopyridin-2-yl) carbamate was synthesized from 2-amino-5-chloronicotinonitrile (500.0mg, 3.26mmol) and reacted with ethylformylhydrazine (150.0mg, 1.79mmol) in the same manner as in example 31(c) to give 9-chloro-2-ethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-ol as a yellow solid (95.0mg, 12%, 2 steps).
LC/MS ESI(+)：250(M+1)，252(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.90(s，1H)，8.72(d，1H，J＝2.7Hz)，8.59(d，1H，J＝2.7Hz)，2.86(q，2H，J＝7.5Hz)，1.35(t，3H，J＝7.5Hz)
(b) Synthesis of 5, 9-dichloro-2-ethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
Reacting 9-chloro-2-ethylpyrido [3,2-e ]][1,2,4]Triazolo [1,5-c]Pyrimidin-5-ol (92.0mg, 0.37mmol) in POCl3(1.5mL), to which was added DIPEA (130.0. mu.L, 0.74 mmol). The reaction mixture was refluxed for 12 hours and then cooled to room temperature. Triethylbenzylammonium chloride (92.0mg, 0.42mmol) was added to the reaction mixture, stirred at 130 ℃ for 2 hours, and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3Aqueous solution was neutralized and then extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM). Collecting the product-containing fractions, and evaporating to obtain 5, 9-dichloro-2-ethylpyrido [3,2-e ] as a pale yellow solid][1,2,4]Triazolo [1,5-c]Pyrimidine (21.0mg, 21%).
LC/MS ESI(+)：268(M+1)，270(M+3)
1H-NMR(300MHz，CDCl3)；：9.12(d，1H，J＝2.7Hz)，8.96(d，1H，J＝2.7Hz)，2.99(q，2H，J＝7.5Hz)，1.35(t，3H，J＝7.5Hz)
(c) Synthesis of 9-chloro-2-ethyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine
5, 9-dichloro-2-ethylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (10.0mg, 0.04mmol) and N-methylpiperazine (8.3. mu.L, 0.08mmol) were reacted in DCM in the same manner as in example 31(e) to give 9-chloro-2-ethyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine as a white solid (5.2mg, 42%).
LC/MS ESI(+)：332(M+1)，334(M+3)
1H-NMR(300MHz，CDCl3)；：8.84(d，1H，J＝2.7Hz)，8.64(d，1H，J＝2.7Hz)，4.13(m，4H)，2.91(q，2H，J＝7.5Hz)，2.53(m，4H)，2.24(s，3H)，1.36(t，3H，J＝7.5Hz)
Example 42
Synthesis of 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-c ] pyrimidine
(a) Synthesis of 6-chloropyrido [2,3-d ] pyrimidine-2, 4(1H,3H) -dione
A mixture of 2-amino-5-chloronicotinamide (169.0mg, 0.98mmol) and diphosgene (646.0mg, 3.14mmol) dissolved in 1, 4-dioxane (10.0mL) was stirred at 120 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, and drying under reduced pressure to obtain a milky white solid compound 6-chloropyrido [2,3-d]Pyrimidine-2, 4(1H,3H) -dione (163.0mg, 84%).
1H-NMR(300MHz，DMSO-d6)；：11.59(s，2H)，8.60(d，1H，J＝2.8Hz)，8.19(d，1H，J＝2.6Hz)
(b) Synthesis of 2,4, 6-trichloropyrido [2,3-d ] pyrimidine
Reacting 6-chloropyrido [2,3-d ]]A mixture of pyrimidine-2, 4(1H,3H) -dione (163.0mg, 0.82mmol) and BPOD (2.0mL) at 180 deg.CStirred for 6 hours and then cooled to room temperature. The reaction mixture was poured into water and saturated NaHCO3The aqueous solution was neutralized (pH 7) and then extracted with EtOAc (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain white solid compound 2,4, 6-trichloropyrido [2,3-d]Pyrimidine (169.0mg, 88%).
1H-NMR(300MHz，DMSO-d6)；：8.97(d，1H，J＝2.8Hz)，8.49(d，1H，J＝2.8Hz)
(c) Synthesis of 2, 6-dichloropyrido [2,3-d ] pyrimidin-4-ol
2,4, 6-trichloropyrido [2,3-d ]]Pyrimidine (90.0mg, 0.38mmol) was dissolved in 1n naoh/BuOH (0.8mL/0.8mL), stirred at room temperature for 2 hours, then distilled under reduced pressure, neutralized with 1n hcl aqueous solution (pH 7), and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and reducing pressure to obtain milky white solid compound 2, 6-dichloropyrido [2,3-d]Pyrimidin-4-ol (63.0mg, 76%).
LC/MS ESI(+)：216(M+1)，218(M+3)
(d) Synthesis of 6-chloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d ] pyrimidin-4 (3H) -one
2, 6-dichloropyrido [2,3-d ]]A mixture of pyrimidin-4-ol (63.0mg, 0.29mmol) and N-methylpiperazine (0.5mL) in EtOH (5.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature and distilled under reduced pressure. The reaction mixture was poured into water and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain milky white solid compound 6-chloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d]Pyrimidin-4 (3H) -one (63.0mg, 77%).
LC/MS ESI(+)：280(M+1)，282(M+3)
(e) Synthesis of 4, 6-dichloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d ] pyrimidine
Reacting 6-chloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d]Pyrimidin-4 (3H) -one (63.0mg, 0.23mmol) and POCl3(1.0mL) of the mixture was stirred at 90 ℃ for 12 hours, followed by distillation under reduced pressure to give 4, 6-dichloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d as a brown solid compound]A pyrimidine.
LC/MS ESI(+)：298(M+1)，300(M+3)
(f) Synthesis of 6-chloro-4-hydrazino-2- (4-methylpiperazin-1-yl) pyrido [2,3-d ] pyrimidine
Unpurified 4, 6-dichloro-2- (4-methylpiperazin-1-yl) pyrido [2,3-d ] pyrimidine, TEA (233.0mg, 2.30mmol) and hydrazine monohydrate (115.0mg, 2.30mmol) were dissolved in EtOH (10.0 mL). The mixture was stirred at-20 ℃ for 1 hour, then distilled under reduced pressure to give 6-chloro-4-hydrazino-2- (4-methylpiperazin-1-yl) pyrido [2,3-d ] pyrimidine as a yellow solid.
LC/MS ESI(+)：294(M+1)，296(M+3)
(g) Synthesis of 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-c ] pyrimidine
Unpurified 6-chloro-4-hydrazino-2- (4-methylpiperazin-1-yl) pyrido [2,3-d]A mixture of pyrimidine and trimethyl orthoformate (2.0mL) was stirred at 100 ℃ for 2 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed to obtain a purple solid compound 9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e][1,2,4]Triazolo [4,3-c]Pyrimidine (0.1mg, 0.1%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.54(s，1H)，8.82(s，1H)，8.76(s，1H)，3.66(bs，4H)，2.55(m，4H)，2.26(s，3H)
Example 43
Synthesis of 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine
(a) Synthesis of ethyl 2- (5-chloro-1H-pyrrolo [2,3-b ] pyridin-3-yl) -2-oxoacetate
Reacting 5-chloro-1H-pyrrolo [2,3-b ]]Pyridine (500.0mg, 3.28mmol) and AlCl3(2180.0mg, 16.38mmol) was dissolved in DCM (10.0mL) to which ethyl 2-chloro-2-oxoacetate (44.0. mu.L, 3.93mmol) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours. EtOH and ice were added to the reaction mixture at 0 ℃ and then extracted with DCM, the organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain yellow solid compound 2- (5-chloro-1H-pyrrolo [2, 3-b)]Pyridin-3-yl) -2-oxoacetic acid ethyl ester (200.0mg, 23%).
LC/MS ESI(+)：253(M+1)，255(M+3)
1H-NMR(300MHz，CDCl3)；：10.39(s，1H)，8.72(d，1H，J＝2.4Hz)，8.68(d，1H，J＝3Hz)，8.39(d，1H，J＝2.4Hz)，4.44(q，2H，J＝7.2Hz)，1.45(t，3H，J＝6.9Hz)
(b) Synthesis of 8-chloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridin-4 (5H) -one
2- (5-chloro-1H-pyrrolo [2, 3-b)]Pyridin-3-yl) -2-oxoacetic acid ethyl ester (200.0mg, 0.79mmol), methylhydrazine sulfate (228.0mg, 1.58mmol) and acetic acid (68.0. mu.L, 11.87mmol) were dissolved in EtOH (2.0mL) and allowed to react in a microwave reactor at 100W at 120 ℃ for 30 minutes and then cooled to room temperature. Then adding NH thereto4Aqueous Cl, extracted with DCM. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered, then distilled under reduced pressure, and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to give the compound 8-chloro-2-methyl-2H-pyrazolo [3,4-c as a cream white solid][1,8]Naphthyridin-4 (5H) -one (30.0mg, 16%).
LC/MS ESI(+)：235(M+1)，237(M+3)
1H-NMR(300MHz，CDCl3)；：11.88(s，1H)，8.71(s，1H)，8.52(d，1H，J＝2.7Hz)，8.38(d，1H，J＝2.4Hz)，4.14(s，3H)
(c) Synthesis of 4, 8-dichloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridine
Reacting 8-chloro-2-methyl-2H-pyrazolo [3,4-c][1,8]Naphthyridin-4 (5H) -one (30.0mg, 0.13mmol) in POCl3(1.5mL) to which DIPEA (66.0. mu.L, 0.38mmol) was then added. The reaction mixture was refluxed for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3The aqueous solution was neutralized (pH 7) and then extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and concentrating under reduced pressure to obtain solid, compound 4, 8-dichloro-2-methyl-2H-pyrazolo [3, 4-c)][1,8]Naphthyridine (30.0mg, 90%).
LC/MS ESI(+)：253(M+1)，255(M+3)
(d) Synthesis of 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine
4, 8-dichloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridine (10.0mg, 0.04mmol) was dissolved in DMF (0.3mL), to which was added N-methylpiperazine (23.3. mu.L, 0.21mmol) and TEA (58.0. mu.L, 0.42 mmol). The reaction mixture was stirred at room temperature for 1 hour and purified by silica gel column chromatography (MeOH: DCM ═ 1: 9). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine as an off-white solid (3.7mg, 30%).
LC/MS ESI(+)：317(M+1)，319(M+3)
1H-NMR(300MHz，CDCl3)；：8.55(s，1H)，8.11(s，1H)，7.99(d，1H，J＝2.4Hz)，4.46(m，4H)，4.22(s，3H)，2.60(m，4H)，2.36(s，3H)
Example 44
Synthesis of 1- (8-chloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine
4, 8-dichloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridine (10.0mg, 0.04mmol) was dissolved in DMF (0.3mL), to which was added azetidin-3-yl (methyl) carbamic acid tert-butyl ester (39.1mg, 0.21mmol) and TEA (58.0. mu.L, 0.42 mmol). The reaction mixture was stirred at room temperature for 1 hour, concentrated under reduced pressure, dissolved in DCM (1.0mL) without purification, and TFA (0.2mL) was added thereto. The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to give 1- (8-chloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine as an off-white solid (3.2mg, 25%).
LC/MS ESI(+)：303(M+1)，305(M+3)
1H-NMR(300MHz，CDCl3+MeOH-d4)；：8.47(s，1H)，8.27(s，1H)，8.13(d，1H，J＝2.4Hz)，4.89(m，2H)，4.71(m，2H)，4.23(s，3H)，3.99(m，1H)，2.76(s，3H)。
Example 45
Synthesis of 8-chloro-2-methyl-4- (piperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine
4, 8-dichloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridine (10.0mg, 0.04mmol) was dissolved in DMF (0.3mL), to which was added piperazine-1-carboxylic acid tert-butyl ester (39.1mg, 0.21mmol) and TEA (58.0. mu.L, 0.42 mmol). The reaction mixture was stirred at room temperature for 1 hour, concentrated under reduced pressure, dissolved in DCM (1.0mL) without purification, and TFA (0.2mL) was added thereto. The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to give the compound 8-chloro-2-methyl-4- (piperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine as a cream-white solid (2.0mg, 16%).
LC/MS ESI(+)：303(M+1)，305(M+3)
1H-NMR(300MHz，CDCl3)；：8.50(d，1H，J＝2.7Hz)，8.16(s，1H)，8.04(d，1H，J＝2.7Hz)，4.52(m，4H)，4.20(s，3H)，3.17(m，4H)。
Example 46
Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (500.0mg, 2.13mmol) and TEA (3.1mL, 21.30mmol) were dissolved in DCM (15.0mL) and N-methylpiperazine (0.3mL, 2.34mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours and poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 20: 1). Collecting the product-containing fractions, and evaporating to obtain 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid]Pyrazine (331.0mg, 52%).
LC/MS ESI(+)：298(M+1)，300(M+3)
(b) Synthesis of 7-chloro-N- (2, 2-diethoxyethyl) -3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazin-2-amine
2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazine (100.0mg, 0.34mmol) and 2, 2-diethoxyethylamine (2.0mL) was stirred at room temperature for 2 hours. The reaction mixture was poured into water and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and distilling under reduced pressure to obtain 7-chloro-N- (2, 2-diethoxyethyl) -3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] as yellow solid compound]Pyrazin-2-amines。
LC/MS ESI(+)：395(M+1)，397(M+3)
(c) Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine
Unpurified 7-chloro-N- (2, 2-diethoxyethyl) -3- (4-methylpiperazin-1-yl) pyrido [2,3-b]A mixture of pyrazin-2-amine and TsOH (128.0mg, 0.68mmol) dissolved in IPA (5.0mL) was stirred at 100 ℃ for 2h, then cooled to room temperature and distilled under reduced pressure. The reaction mixture was poured into DCM and extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by column chromatography over silica gel (DCM: MeOH ═ 20:1 to 90: 10). Collecting the product-containing fractions, and evaporating to obtain 8-chloro-4- (4-methylpiperazin-1-yl) imidazo [1,2-a as a yellow solid]Pyrido [2,3-e ]]Pyrazine (56.0mg, 55%, 2 steps).
LC/MS ESI(+)：303(M+1)，305(M+3)
1H-NMR(300MHz,DMSO-d6)；：8.81(d，1H，J＝2.4Hz)，8.74(d，1H，J＝1.2Hz)，8.53(d，1H，J＝2.4Hz)，7.72(d，1H，J＝1.4Hz)，4.42(bs，4H)，2.49(m，4H)，2.23(s，3H)
Example 47
Synthesis of 8-chloro-4- (piperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine
(a) Synthesis of tert-butyl 4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (400.0mg, 1.71mmol) and TEA (2.4mL, 17.10mmol) were dissolved in DCM (8.0mL), to which was slowly added tert-butyl piperazine-1-carboxylate (350.0mg, 1.88mmol) diluted with DCM (2.0mL) at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). Is washed with brineOrganic layer of anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 19: 1). Collecting the product-containing fractions, and evaporating to obtain 4- (2, 7-dichloropyrido [2,3-b ] as yellow solid]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：384(M+1)，386(M+3)
(b) Synthesis of tert-butyl 4- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) piperazine-1-carboxylate
Unpurified 4- (2, 7-dichloropyrido [2,3-b ]]A mixture of pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester and 2, 2-diethoxyethylamine (2.0mL) was stirred at room temperature for 12 hours, poured into water, and then extracted with EtOAc (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain yellow solid compound 4- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]Pyrazin-3-yl) piperazine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：481(M+1)，483(M+3)
(c) Synthesis of 8-chloro-4- (piperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine
Unpurified 4- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) piperazine-1-carboxylate and TsOH (662.0mg, 3.48mmol) dissolved in IPA (10.0mL) was stirred at 100 deg.C for 1 hour, then cooled to room temperature and distilled under reduced pressure. The reaction mixture was poured into water and washed with NaHCO3The aqueous solution was neutralized (pH 7) and then extracted with DCM (50.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 20:1 to 90: 10). Collecting the product-containing fractions, and evaporating to obtain 8-chloro-4- (piperazin-1-yl) imidazo [1,2-a as a yellow solid]Pyrido [2,3-e ]]Pyrazine (155.0mg, 31%, 2 steps).
LC/MS ESI(+)：289(M+1)，291(M+3)
1H-NMR(300MHz,DMSO-d6)；：8.79(d，1H，J＝2.5Hz)，8.74(m，1H)，8.52(d，1H，J＝2.4Hz)，7.72(m，1H)，4.35(bs，4H)，2.84(m，4H)
Example 48
Synthesis of 1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (50.0mg, 0.11mmol), N' -dimethylethane-1, 2-diamine (1.0mg, 0.01mmol), CuI (1.1mg, 5.50 μmol), and KI (38.0mg, 0.22mmol) were added to 1-butanol (1.0mL), stirred at 100 ℃ for 18 hours, distilled under reduced pressure, and then purified by silica gel column chromatography (DCM: MeOH 98:2-95: 5). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (25.0mg, 45%) as a white solid.
LC/MS ESI(+)：482(M+1)
(b) Synthesis of 1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
1- (8-iodopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (25.0mg, 0.05mmol) was dissolved in DCM (3.0mL) to which TFA (0.5mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour and distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and extracted with DCM/MeOH (9/1, 30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, reducingDistillation under pressure, followed by purification by amine silica gel column chromatography (DCM: MeOH ═ 96: 4). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-iodopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (15.0mg, 75%).
LC/MS ESI(+)：382(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.96(s，1H)，8.97(d，1H，J＝2.1Hz)，8.68(d，1H，J＝2.1Hz)，4.89(m，1H)，4.42(m，2H)，4.00(m，1H)，3.70(m，1H)，2.29(s，3H)
Example 49
Synthesis of 8-iodo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 5-iodopyridine-2, 3-diamine
5-iodo-3-nitropyridin-2-amine (2000.0mg, 7.55mmol), Fe (4800.0mg, 85.90mmol), and concentrated HCl (0.1mL) were added to a mixture of EtOH (7.0mL) and water (2.0mL), and the suspension was stirred at 100 ℃ for 30. The reaction mixture was cooled to room temperature, filtered through celite, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). Fractions containing the product were collected and evaporated to give 5-iodopyridine-2, 3-diamine (1710.0mg, 97%) as a brown solid.
LC/MS ESI(+)：236(M+1)
1H-NMR(300MHz，DMSO-d6)；：7.37(d，1H，J＝2.1Hz)，6.91(d，1H，J＝2.1Hz)，5.59(s，2H)，4.91(s，2H)
(b) Synthesis of 2, 3-dichloro-7-iodopyrido [2,3-b ] pyrazine
A mixture of 5-iodopyridine-2, 3-diamine (1400.0mg, 5.96mmol) and diethyl oxalate (10.0mL) was stirred at 100 ℃ for 12 hours and then cooled to room temperature. Adding Et2O is added to the reaction mixtureTo form a solid, and then filtering the formed solid, drying under reduced pressure to obtain a brown solid compound 7-iodopyrido [2,3-b]Pyrazine-2, 3-diols. Unpurified 7-iodopyrido [2,3-b ]]Pyrazine-2, 3-diols and POCl3(15.0mL) of the mixture was stirred at 150 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water, and the formed solid was filtered and then dried under reduced pressure to give 2, 3-dichloro-7-iodopyrido [2,3-b ] as a brown solid compound]Pyrazine (1080.0mg, 56%).
LC/MS ESI(+)：326(M+1)，328(M+3)
(c) Synthesis of 2-chloro-7-iodo-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2, 3-dichloro-7-iodopyrido [2,3-b ] pyrazine (50.0mg, 0.15mmol) and TEA (213.0. mu.L, 1.53mmol) were dissolved in DCM (2.0mL) and N-methylpiperazine (16.0. mu.L, 0.15mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 hour, concentrated, and then purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound 2-chloro-7-iodo-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (40.0mg, 67%) as a yellow solid.
LC/MS ESI(+)：390(M+1)，392(M+3)
(d) Synthesis of 2-hydrazino-7-iodo-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2-chloro-7-iodo-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (40.0mg, 0.10mmol) and hydrazine monohydrate (7.5 μ L, 0.15mmol) were dissolved in EtOH (1.0mL) and stirred at room temperature for 3 hours to form a solid. The solid formed was filtered to give the compound 2-hydrazino-7-iodo-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (39.4mg, 100%) as a yellow solid.
LC/MS ESI(+)：386(M+1)
(e) Synthesis of 8-iodo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
2-hydrazino-7-iodo-3- (4-methylpiperazin-1-yl)) Pyrido [2,3-b ]]A mixture of pyrazine (39.4mg, 0.10mmol) and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 3 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain 8-iodo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazine (13.5mg, 34%).
LC/MS ESI(+)：396(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.00(s，1H)，9.00(d，1H，J＝2.1Hz)，8.71(d，1H，J＝2.1Hz)，4.57-4.17(m，4H)，2.52(m，4H)，2.24(s，3H)
Example 50
Synthesis of N-methyl-1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
(a) Synthesis of 5-methylpyridine-2, 3-diamine
5-methyl-3-nitropyridin-2-amine (50000.0mg, 0.33mol) was dissolved in MeOH/EtOAc (1000.0mL) to which was added 10% Pd/C (5000.0 mg). The flask was replaced with hydrogen. The reaction mixture was reacted at room temperature for 8 hours, filtered through celite, and evaporated under reduced pressure to give 5-methylpyridine-2, 3-diamine (98000.0mg, 82%) as a brown solid.
1H-NMR(300MHz，CDCl3)；：7.48(s，1H)，6.74(s，1H)，4.06(bs，2H)，3.27(bs，2H)，2.16(s，3H)
(b) Synthesis of 7-methylpyrido [2,3-b ] pyrazine-2, 3-diol
5-methylpyridine-2, 3-diamine (98000.0mg, 0.80mol) and oxalic acid (76900.0mg, 0.85mol) were added to 3N HCl (784.0 mL). The mixture was stirred at 120 ℃ for 24 hours and then cooled to room temperature. The resulting solid was filtered and dried under reduced pressure to give 7-methylpyrido [2,3-b ] pyrazine-2, 3-diol (140000.0mg, 100%) as a brown solid.
1H-NMR(300MHz，DMSO-d6)；：12.26(bs，1H)，11.97(bs，1H)，7.90(s，1H)，7.25(s，1H)，2.25(s，3H)
(c) Synthesis of 2, 3-dichloro-7-methylpyrido [2,3-b ] pyrazine
Reacting 7-methylpyrido [2,3-b ]]Pyrazine-2, 3-diol (140000.0mg, 0.79mol) and POCl3(900.0mL) of the mixture was stirred at 130 ℃ for 24 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid. The solid formed was filtered, dried under reduced pressure and then purified by silica gel column chromatography (DCM). Collecting the product-containing fractions, and evaporating to obtain 2, 3-dichloro-7-methylpyrido [2,3-b ] as a brown solid]Pyrazine (55000.0mg, 33%).
(d) Synthesis of tert-butyl (1- (2-chloro-7-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
2, 3-dichloro-7-methylpyrido [2,3-b ] pyrazine (750.0mg, 3.47mmol) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (850.0mg, 3.82mmol) were reacted in the same manner as in example 2(c) to give the compound (1- (2-chloro-7-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (720.0mg, 57%) as a pale yellow solid.
LC/MS ESI(+)：364(M+1)
1H-NMR(300MHz，CDCl3)；：8.74(d，1H，J＝2.1Hz)，7.91(d，1H，J＝2.1Hz)，5.05(m，1H)，4.71(m，2H)，4.53(m，2H)，2.99(s，3H)，2.51(s，3H)，1.50(s，9H)
(e) Synthesis of methyl (1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester
1- (2-chloro-7-methylpyrido [2,3-b ]]Pyrazin-3-yl) nitrogensTert-butyl azetidin-3-yl) (methyl) carbamate (320.0mg, 0.88mmol) and hydrazine monohydrate (0.1mL, 3.50mmol) were dissolved in EtOH (8.0mL) and stirred at 40 ℃ for 12 h. Et was added thereto2O to form a solid, filtering the solid formed, and drying under reduced pressure to give a dark brown compound (1- (2-hydrazino-7-methylpyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester. The unpurified compound (1- (2-hydrazino-7-methylpyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester and trimethyl orthoformate (4.0mL) were reacted to give methyl (1- (8-methylpyrido [2, 3-e) as a brown solid compound][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (92.0mg, 29%, 2 steps).
LC/MS ESI(+)：370(M+1)
1H-NMR(300MHz，CDCl3)；：9.14(s，1H)，8.52(d，1H，J＝2.1Hz)，7.84(d，1H，J＝2.1Hz)，4.60(m，5H)，2.99(s，3H)，2.46(s，3H)，1.47(s，9H)
(f) Synthesis of N-methyl-1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
Methyl (1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (90.0mg, 0.24mmol) was dissolved in a mixture of DCM (2.0mL) and TFA (3.0 mL). The reaction mixture was stirred at 0 ℃ for 1 hour, then purified by amine silica gel column chromatography (DCM: MeOH ═ 50: 1). The product-containing fractions were collected and evaporated to yield the yellow solid compound N-methyl-1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (39.0mg, 59%).
LC/MS ESI(+)：270(M+1)
1H-NMR(300MHz，CDCl3)；：9.13(s，1H)，8.51(d，1H，J＝2.1Hz)，7.83(d，1H，J＝2.1Hz)，5.09(m，1H)，4.75-4.55(m，2H)，4.20(m，1H)，3.89(m，1H)，2.50(s，6H)
Example 51
Synthesis of 1- (8- (difluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-formylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
2.5M n-BuLi in hexane was added slowly to (1- (8-iodopyrido [2,3-e ]) at-78 deg.C][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (270.0mg, 0.56mmol) and THF (5.6mL) in suspension. The reaction mixture was stirred at-78 ℃ for 10 minutes, to which DMF (0.5mL) and THF (2.0mL) were then added. The reaction mixture was stirred for 1 hour, and NH was added thereto4Aqueous Cl, extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated to give the compound (1- (8-formylpyrido [2, 3-e) as a yellowish white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (18.0mg, 8%).
LC/MS ESI(+)：384(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.14(s，1H)，10.10(s，1H)，9.04(d，1H，J＝2.1Hz)，8.97(d，1H，J＝2.1Hz)，5.09-5.00(m，2H)，4.89(m，1H)，4.60(m，1H)，4.46(m，1H)，2.94(s，3H)，1.42(s，9H)
(b) Synthesis of tert-butyl (1- (8- (difluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
1- (8-formyl pyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (11.0mg, 0.03 mmo)l) and Deoxo-Fluor (0.8mL) were reacted in a microwave reactor at 50W at 70 ℃ for 1.5 hours and then cooled to room temperature. The reaction mixture was poured into ice water, and NH was added thereto4Aqueous Cl solution, extraction with MeOH/DCM ═ 1:20 solution, and extraction with anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1:60), and fractions containing the product were collected and evaporated to give the compound (1- (8- (difluoromethyl) pyrido [2,3-e ] as a cream-white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (5.0mg, 45%).
1H-NMR(300MHz，CDCl3)；：9.23(s，1H)，8.76(s，1H)，8.19(s，1H)，7.04-6.66(m，1H)，5.18(m，1H)，4.99(m，1H)，4.74(m，1H)，4.56(m，1H)，4.35-4.25(m，1H)，3.01(s，3H)，1.49(s，9H)
(c) Synthesis of 1- (8- (difluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (8- (difluoromethyl) pyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (5.0mg, 0.01mmol) and DCM (0.6mL) was stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. Filtering the solid formed, drying under reduced pressure to obtain a milky white solid compound 1- (8- (difluoromethyl) pyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (1.5mg, 41%).
LC/MS ESI(+)：306(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.07(s，1H)，8.83(s，1H)，8.69(s，1H)，7.42-7.05(m，1H)，4.93(m，1H)，4.52-4.40(m，2H)，4.01(m，1H)，3.72(m，1H)，2.30(s，3H)
Example 52
Synthesis of N-methyl-1- (8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
(a) Synthesis of 3-nitro-5- (trifluoromethyl) pyridin-2-amine
5- (trifluoromethyl) pyridin-2-amine (1700.0mg, 10.49mmol) was dissolved in concentrated H2SO4(10.0mL) to which concentrated HNO was then added slowly3(1.7mL, 26.22 mmol). The reaction mixture was stirred at 80 ℃ for 48 hours, then added to ice water, neutralized with 1N aqueous NaOH (pH 9), and then extracted with EtOAc (200.0 mL). The organic layer was washed with brine, Na-free2SO4Water drying, filtering, and then distilling under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 50: 50). The product-containing fractions were collected and evaporated to give 3-nitro-5- (trifluoromethyl) pyridin-2-amine (538.0mg, 25%) as a yellow solid.
LC/MS ESI(+)：208(M+1)
1H-NMR(300MHz，CDCl3)；：8.67(d，1H，J＝1.7Hz)，8.59(d，1H，J＝1.7Hz)，7.92(bs，1H)，6.10(bs，1H)
(b) Synthesis of 5- (trifluoromethyl) pyridine-2, 3-diamine
3-Nitro-5- (trifluoromethyl) pyridin-2-amine (538.0mg, 2.60mmol) and SnCl22H2O (2340.0mg, 10.39mmol) was added to DMF (5.0mL) and stirred at 60 ℃ for 12 h. The reaction mixture was poured into saturated NaHCO3In aqueous solution, neutralized (pH 7), and then extracted with EtOAc (200.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtration and evaporation under reduced pressure gave the compound 5- (trifluoromethyl) pyridine-2, 3-diamine as a brown solid.
LC/MS ESI(+)：178(M+1)
(c) Synthesis of 2, 3-dichloro-7- (trifluoromethyl) pyrido [2,3-b ] pyrazine
Unpurified 5- (trifluoromethyl) pyridine-2, 3-diamine was added to diethyl oxalate (10.0 mL). The mixture was stirred at 120 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid, and filtering the formed solid under reduced pressure to obtain a brown solid compound 7- (trifluoromethyl) pyrido [2,3-b]Pyrazine-2, 3-diols. Unpurified 7- (trifluoromethyl) pyrido [2,3-b ]]Pyrazine-2, 3-diols and POCl3(10.0mL) of the mixture was stirred at 130 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid. The solid formed was filtered and then dried under reduced pressure to give 2, 3-dichloro-7- (trifluoromethyl) pyrido [2,3-b ] as a brown solid]Pyrazine (370.0mg, 53%, 3 steps).
LC/MS ESI(+)：268(M+1)，270(M+3)，272(M+5)
(d) Synthesis of tert-butyl (1- (2-chloro-7- (trifluoromethyl) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
2, 3-dichloro-7- (trifluoromethyl) pyrido [2,3-b]Pyrazine (100.0mg, 0.37mmol) and TEA (0.26mL, 1.85mmol) were dissolved in DCM (10.0mL) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (83.0mg, 0.37mmol) diluted with DCM (5.0mL) and TEA (0.26mL, 1.85mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into saturated NH4Aqueous Cl was extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give (1- (2-chloro-7- (trifluoromethyl) pyrido [2, 3-b) as a brown solid]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：418(M+1)
(e) Synthesis of tert-butyl (1- (2-hydrazino-7- (trifluoromethyl) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
Unpurified (1- (2-chloro-7- (trifluoromethyl) pyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester and hydrazine monohydrate (47.0mg, 0.93mmol) were dissolved in EtOH (5.0mL), stirred at room temperature for 12 hours, and then distilled under reduced pressure. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure to obtain the compound (1- (2-hydrazino-7- (trifluoromethyl) pyrido [2, 3-b) as a yellow solid]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：414(M+1)
(f) Synthesis of methyl (tert-butyl 1- (8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate
Unpurified (1- (2-hydrazino-7- (trifluoromethyl) pyrido [2, 3-b)]A mixture of t-butyl pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate and trimethyl orthoformate (10.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed and then drying under reduced pressure to give methyl (1- (8- (trifluoromethyl) pyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：424(M+1)
(g) Synthesis of N-methyl-1- (8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
The unpurified methyl (1- (8- (trifluoromethyl) pyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester was dissolved in DCM (4.0mL) and TFA (0.5mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was distilled under reduced pressure. With NaHCO3The residue was neutralized with aqueous solution (pH 7) and then extracted with DCM (30.0 mL).The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to give the milky white solid compound N-methyl-1- (8- (trifluoromethyl) pyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-amine (26.7mg, 20%, 4 steps).
LC/MS ESI(+)：324(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.1(s，1H)，9.01(bs，1H)，8.83(bs，1H)，4.95(m，1H)，4.48(m，2H)，4.03(m，1H)，3.74(m，1H)，2.30(s，3H)
Example 53
Synthesis of 4- (4-methylpiperazin-1-yl) -8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2-chloro-3- (4-methylpiperazin-1-yl) -7- (trifluoromethyl) pyrido [2,3-b ] pyrazine
2, 3-dichloro-7- (trifluoromethyl) pyrido [2,3-b]Pyrazine (50.0mg, 0.19mmol) and TEA (0.3mL, 1.90mmol) were dissolved in DCM (5.0mL) and N-methylpiperazine (21.0. mu.L, 0.19mmol) diluted with DCM (5.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 2-chloro-3- (4-methylpiperazin-1-yl) -7- (trifluoromethyl) pyrido [2,3-b ] as a yellow solid]Pyrazine (53.0mg, 85%).
LC/MS ESI(+)：332(M+1)，334(M+3)
(b) Synthesis of 4- (4-methylpiperazin-1-yl) -8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
2-chloro-3- (4-methylpiperazin-1-yl) -7- (trifluoromethyl) pyrido [2,3-b]Pyrazine (53.0mg, 0.16mmol) and hydrazine monohydrate (0.1mL, 2.04mmol) were dissolved in EtOH (2.0mL), stirred at room temperature for 1 hour, and then distilled under reduced pressure. The reaction mixture was poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and drying under reduced pressure to obtain yellow solid compound 2-hydrazino-3- (4-methylpiperazin-1-yl) -7- (trifluoromethyl) pyrido [2,3-b]A pyrazine. Unpurified 2-hydrazino-3- (4-methylpiperazin-1-yl) -7- (trifluoromethyl) pyrido [2,3-b]A mixture of pyrazine and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). Collecting the product-containing fractions, and evaporating to obtain 4- (4-methylpiperazin-1-yl) -8- (trifluoromethyl) pyrido [2,3-e as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazine (14.3mg, 23%, 3 steps).
LC/MS ESI(+)：338(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.13(s，1H)，9.05(d，1H，J＝2.1Hz)，8.87(d，1H，J＝2.1Hz)，5.00～4.00(m，4H)，2.53(m，4H)，2.26(s，3H)
Example 54
Synthesis of 1- (8-ethynylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of methyl (tert-butyl 1- (8- ((trimethylsilyl) ethynyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate
1- (8-bromopyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (A)50.0mg, 0.12mmol), TMS-acetylene (81.0. mu.L, 0.58mmol), PdCl2(PPh3)2(8.1mg, 0.01mmol), CuI (4.4mg, 0.02mmol) and TEA (0.5mL) were added to DMF (0.5mL), and the mixture was reacted in a microwave reactor at 50W at 70 ℃ for 1.5 hours and then cooled to room temperature. DCM and water were added to the reaction mixture to form a solid, and the solid formed was filtered to give a grey solid. The resulting grey solid was stirred in MeOH: DCM ═ 1:9 solution and then filtered. The filtrate was distilled under reduced pressure to give methyl (1- (8- ((trimethylsilyl) ethynyl) pyrido [2, 3-e) as a gray solid compound][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (50.0mg, 96%).
LC/MS ESI(+)：452(M+1)
1H-NMR(300MHz，DMSO-d6)：10.00(s，1H)，8.75(d，1H，J＝2.1Hz)，8.57(d，1H，J＝2.1Hz)，5.25-4.85(m，2H)，4.82(m，1H)，4.51(m，1H)，4.41(m，1H)，2.93(s，3H)，1.42(s，9H)，0.27(s，9H)
(b) Synthesis of 1- (8-ethynylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
Methyl (tert-butyl 1- (8- ((trimethylsilyl) ethynyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate (50.0mg, 0.11mmol) was dissolved in 4N HCl in dioxane (1.0 mL). The reaction mixture was stirred at room temperature for 1 hour to form a solid, and the formed solid was filtered. The resulting solid was dissolved in THF (1.0mL), to which was added TBAF (0.2mL, 0.22 mmol). The reaction mixture was stirred at room temperature for 1 hour, to which MeOH was added to form a solid. The solid formed was filtered and then dried under reduced pressure to give 1- (8-ethynylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine as a cream white solid (6.0mg, 19%).
LC/MS ESI(+)：280(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.99(s，1H)，8.76(d，1H，J＝1.8Hz)，8.60(d，1H，J＝1.8Hz)，4.94(m，1H)，4.60-4.40(m，3H)，4.10(m，1H)，3.81(m，1H)，3.16(m，1H)，2.36(s，3H)
Example 55
Synthesis of N-methyl-1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
(a) Synthesis of methyl (1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester
1- (8-bromopyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (150.0mg, 0.35mmol), tributylvinyltin (1.0mL, 3.45mmol), Pd (OAc)2(15.0mg，0.07mmol)、CuI(39.4mg，0.21mmol)、P(o-Tol)3(42.0mg, 0.14mmol) and TEA (240.0. mu.L, 1.73mmol) were added to CH3CN (2.5 mL). The mixture was reacted in a microwave reactor at 100W at 100 ℃ for 3 hours and then cooled to room temperature. The reaction mixture was purified by silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated to give methyl (1- (8-vinylpyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (15.0mg, 11%).
LC/MS ESI(+)：382(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.00(s，1H)，8.78(s，1H)，8.62(bs，1H)，6.86(m，1H)，6.06(m，1H)，5.45(m，1H)，5.20-4.65(m，3H)，4.65-4.20(m，2H)，2.93(s，3H)，1.42(s，9H)
(b) Synthesis of N-methyl-1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
Methyl (tert-butyl 1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate (15.0mg, 0.02mmol) was dissolved in DCM (0.6mL), to which TFA (0.4mL) was then added. The reaction mixture was stirred at room temperature for 1 hour, then purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give the compound N-methyl-1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine as a cream-white solid (1.5mg, 29%).
LC/MS ESI(+)：282(M+1)
1H-NMR(300MHz，DMSO-d6)；：9.99(s，1H)，8.76(s，1H)，8.60(s，1H)，6.86(m，1H)，6.06(m，1H)，5.45(m，1H)，4.90(m，1H)，4.55-4.25(m，2H)，3.99(m，1H)，3.71(m，1H)，2.29(s，3H)
Example 56
Synthetic amines of 1- (8-ethylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-
Methyl tert-butyl (1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate (12.0mg, 0.03mmol) was dissolved in MeOH (1.0mL), to which 10% Pd/C (4.8mg) was added. The flask was replaced with hydrogen gas, and then allowed to react for 12 hours. The reaction mixture was filtered through celite, and then distilled under reduced pressure. The residue was dissolved in DCM (0.6mL), and TFA (0.4mL) was added thereto. The reaction mixture was stirred for 1 hour, then purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give 1- (8-ethylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine as a cream solid (3.5mg, 39%).
LC/MS ESI(+)：284(M+1)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.52(d，1H，J＝2.1Hz)，7.82(d，1H，J＝2.1Hz)，5.09(m，1H)，4.78-4.50(m，2H)，4.22(m，1H)，3.85(m，1H)，2.80(q，2H，J＝7.5Hz)，2.49(s，3H)，1.35(t，3H，J＝7.5Hz)
Example 57
Synthesis of 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-ol
1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (38.0mg, 0.13mmol) was dissolved in DCM (3.0mL) and then cooled to-78 ℃. To this was added 1M tribromoborane (3.0mL) dissolved in DCM at a concentration of 1M. The temperature was gradually increased to 40 ℃. The reaction mixture was stirred for 12 hours, then cooled to-78 ℃ and MeOH (3.0mL) was added. The reaction mixture was allowed to warm to room temperature. The reaction mixture was then concentrated and purified by amine silica gel column chromatography (DCM: MeOH ═ 95:5-90: 10). The product-containing fractions were collected and evaporated to yield the compound 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-ol as a white solid (18.0mg, 50%).
LC/MS ESI(+)：272(M+1)
1H-NMR(300MHz，MeOH-d4)；：9.73(s，1H)，8.20(m，1H)，7.90(m，1H)，4.81(m，2H)，4.45(m，2H)，4.01(m，1H)，2.59(s，3H)
Example 58
Synthesis of 1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
A mixture comprising MeOH (4.5mL) and NaH (450.0mg) was stirred at room temperature for 30 minutes, then added to a solution of tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (300.0mg, 0.69mmol) and CuI (262.8mg, 1.38mmol) in DMF (4.5 mL). The reaction mixture was reacted in a microwave reactor at 60W at 90 ℃ for 30 minutes, cooled to room temperature, and then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (49.0mg, 55%) as a white solid.
LC/MS ESI(+)：386(M+1)
(b) Synthesis of 1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (49.0mg, 0.13mmol) was dissolved in DCM (0.6mL), to which TFA (0.4mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour, then concentrated and purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield 1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine as a white solid (25.0mg, 33%).
LC/MS ESI(+)：286(M+1)
1H-NMR(300MHz，MeOH-d4)；：9.82(s，1H)，8.30(d，1H，J＝2.7Hz)，8.15(d，1H，J＝2.7Hz)，4.76-4.19(m，4H)，4.00(s，3H)，3.83(m，1H)，2.43(s，3H)
Example 59
Synthesis of 1- (8- (difluoromethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8- (benzyloxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Tert-butyl (1- (8- (benzyloxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (200.0mg, 0.60mmol), 1, 10-phenanthroline (21.6mg, 0.12mmol), cesium carbonate (391.0mg, 1.20mmol), and CuI (114.3mg, 0.60mmol) were dissolved in benzyl alcohol (4.0 mL). The mixture was reacted in a microwave reactor at 100W at 100 ℃ for 2 hours, then cooled to room temperature, and 36.0mL of water was added thereto. The reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8- (benzyloxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (120.0mg, 43%) as a white solid.
LC/MS ESI(+)：462(M+1)
(b) Synthesis of tert-butyl (1- (8-hydroxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Tert-butyl (1- (8- (benzyloxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (120.0mg, 0.32mmol) and 10% Pd/C (30.0mg) were dissolved in MeOH (10.0 mL). The flask was replaced with hydrogen, and then stirred at room temperature for 3 days. The reaction mixture was filtered through celite, distilled under reduced pressure, and then purified by column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8-hydroxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (40.0mg, 34%) as a white solid.
LC/MS ESI(+)：372(M+1)
(c) Synthesis of 1- (8- (difluoromethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
1- (8-hydroxypyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (20.0mg, 0.05mmol) in CH3CN (0.7mL), to which was added KOH (60.0mg, 1.08mmol) dissolved in water (0.7mL), followed by diethyl (bromodifluoromethyl) phosphonate (60.0. mu.L, 0.32 mmol). The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, dissolved without further purification in DCM (1.0mL), and then TFA (0.2mL) was added thereto. The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain 1- (8- (difluoromethoxy) pyrido [2,3-e ] as a white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (10.0mg, 17%).
LC/MS ESI(+)：322(M+1)
1H-NMR(300MHz，MeOH-d4)；：9.79(s，1H)，8.41(m，2H)，7.24-6.75(m，1H)，5.03(m，1H)，4.57(m，2H)，4.15(m，1H)，3.82(m，1H)，2.43(s，3H)
Example 60
Synthesis of 8-chloro-7-methoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (10.0mg, 0.03mmol) and NaH (4.0 mg: in 60% oil, 0.12mmol) were dissolved in MeOH (1.0 mL). The mixture was reacted in a microwave reactor at 60W at 90 ℃ for 1 hour. The reaction mixture was concentrated under reduced pressure and then purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). The product-containing fractions were collected and evaporated to yield 8-chloro-7-methoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream-white solid (3.7mg, 37%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.91(s，1H)，8.85(s，1H)，4.41(m，4H)，4.01(m，3H)，2.50(m，4H)，2.26(s，3H)
Example 61
Synthesis of 8-chloro-7-methoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Tert-butyl 4- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate (20.0mg, 0.05mmol) and NaH (9.0 mg: 60% in oil, 0.25mmol) obtained in the same manner as in examples 2(c) - (e) were dissolved in MeOH (1.0 mL). The mixture was stirred in a microwave reactor at 60W at 90 ℃ for 1 hour and then distilled under reduced pressure. The reaction mixture was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give the compound 4- (8-chloro-7-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylic acid tert-butyl ester as a cream white solid. Tert-butyl 4- (8-chloro-7-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate was dissolved in DCM (2.0mL), and TFA (0.5mL) was slowly added thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, and then distilled under reduced pressure. The residue was neutralized with TEA (pH 7) and then purified by amine silica gel column chromatography (DCM: MeOH 99: 1). The product-containing fractions were collected and evaporated to give the compound 8-chloro-7-methoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream-white solid (9.0mg, 60%, 2 steps).
LC/MS ESI(+)：320(M+1)，322(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.98(s，1H)，8.92(s，1H)，4.40(m，4H)，4.08(s，3H)，2.94(m，4H)
Example 62
Synthesis of 7, 8-dichloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Tert-butyl 4- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate (20.0mg, 0.05mmol) was dissolved in DCM (2.0mL), to which TFA (0.5mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and distilled under reduced pressure. The residue was neutralized with TEA (pH 7) and purified by amine silica gel column chromatography (DCM: MeOH 99: 1). The product-containing fractions were collected and evaporated to yield the compound 7, 8-dichloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream-white solid (14.0mg, 92%).
LC/MS ESI(+)：324(M+1)，326(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，9.08(s，1H)，5.00-4.00(m，4H)，2.95(m，4H)
Example 63
Synthesis of 8-chloro-7-ethoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Tert-butyl 4- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate (20.0mg, 0.05mmol) and NaOEt (16.0mg, 0.25mmol) were dissolved in EtOH (1.0 mL). The mixture was reacted in a microwave reactor at 60W at 90 ℃ for 1 hour, cooled to room temperature, distilled under reduced pressure, and then purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). The product-containing fractions were collected and evaporated to give the compound 4- (8-chloro-7-ethoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylic acid tert-butyl ester as a cream-white solid. Tert-butyl 4- (8-chloro-7-ethoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) piperazine-1-carboxylate was dissolved in DCM (2.0mL), to which TFA (0.5mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours and distilled under reduced pressure. The residue was neutralized with TEA (pH 7) and then purified by amine silica gel column chromatography (DCM: MeOH 99: 1). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-7-ethoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream-white solid (4.0mg, 25%, 2 steps).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.98(m，1H)，8.91(m，1H)，4.53(q，2H，J＝7.1Hz)，4.40(m，4H)，2.95(m，4H)，1.46(t，3H，J＝7.1Hz)
Example 64
Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) -7- (2,2, 2-trifluoroethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Mixing 7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (10.0mg, 0.03mmol) and NaH (12.0 mg: in 60% oil, 0.30mmol) were dissolved in CF3CH2OH (1.0 mL). The mixture was then reacted in a microwave reactor at 60W at 120 ℃ for 1 hour, distilled under reduced pressure, and then purified by amine silica gel column chromatography (DCM: MeOH ═ 99: 1). The product-containing fractions were collected and evaporated to give the compound 8-chloro-4- (4-methylpiperazin-1-yl) -7- (2,2, 2-trifluoroethoxy) pyrido [2,3-e as a cream-white solid][1,2,4]Triazolo [4,3-a]Pyrazine (3.51mg, 29%).
LC/MS ESI(+)：402(M+1)，404(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.93(m，1H)，8.93(m，1H)，5.14(q，2H，J＝8.9Hz)，4.43(m，4H)，2.50(m，4H)，2.25(s，3H)
Example 65
Synthesis of 1- (8-bromo-9-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of 5-bromo-4-methylpyridine-2, 3-diamine
5-bromo-4-methyl-3-nitropyridin-2-amine (700.0mg, 3.02mmol), Fe (1680.0mg, 30.20mmol) and concentrated HCl (50.0 μ L) were added to EtOH (2.8mL) and water (0.7 mL). The suspension was stirred at 100 ℃ for 30 minutes. The reaction mixture was cooled to room temperature, filtered through celite, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). Fractions containing the product were collected and evaporated to give 5-bromo-4-methylpyridine-2, 3-diamine (550.0mg, 90%) as a brown solid.
LC/MS ESI(+)：202(M+1)，204(M+3)
1H-NMR(300MHz，DMSO-d6)；：7.37(s，1H)，5.49(s，2H)，4.74(s，2H)，2.12(s，3H)
(b) Synthesis of 7-bromo-8-methylpyrido [2,3-b ] pyrazine-2, 3-diol
A mixture of 5-bromo-4-methylpyridine-2, 3-diamine (550.0mg, 2.72mmol) and diethyl oxalate (10.0mL) was stirred at 100 ℃ for 12 hours and then cooled to room temperature. Adding Et2O is added to the reaction mixture to form a solid, the solid formed is filtered and dried under reduced pressure to obtain the brown solid compound 7-bromo-8-methylpyrido [2,3-b]Pyrazine-2, 3-diol (595.0mg, 85%).
LC/MS ESI(+)：256(M+1)，258(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.40(s，1H)，11.57(s，1H)，8.19(s，1H)，2.45(s，3H)
(c) Synthesis of 7-bromo-2, 3-dichloro-8-methylpyrido [2,3-b ] pyrazine
Reacting 7-bromo-8-methylpyrido [2,3-b ]]Pyrazine-2, 3-diol (560.0mg, 2.19mmol) and POCl3(5.0mL) of the mixture was stirred at 100 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid. The solid formed was filtered and dried under reduced pressure. The resulting solid was dissolved in DCM and passed throughPurification by silica gel column chromatography (EtOAc: n-Hex ═ 1: 9). Collecting the product-containing fractions, and evaporating to obtain 7-bromo-2, 3-dichloro-8-methylpyrido [2,3-b ] as a red solid]Pyrazine (365.0mg, 54%).
LC/MS ESI(+)：292(M+1)，294(M+3)，296(M+5)
1H-NMR(300MHz，DMSO-d6)；：9.25(s，1H)，2.78(s，3H)
(d) Synthesis of tert-butyl (1- (7-bromo-2-chloro-8-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
TEA (72.0. mu.L, 0.51mmol) was added to a mixture of 7-bromo-2, 3-dichloro-8-methylpyrido [2,3-b ] pyrazine (50.0mg, 0.17mmol), azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (40.0mg, 0.18mmol) and DCM (1.7mL) at 0 ℃ and stirred for 1 h. The reaction mixture was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (7-bromo-2-chloro-8-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (58.0mg, 76%) as a yellow solid.
LC/MS ESI(+)：442(M+1)，444(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.85(s，1H)，4.86(m，1H)，4.65-4.55(m，2H)，4.49-4.40(m，2H)，2.90(s，3H)，2.66(s，3H)，1.41(s，9H)
(e) Synthesis of tert-butyl (1- (8-bromo-9-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Hydrazine monohydrate (20.0. mu.L, 0.63mmol) was added to (1- (7-bromo-2-chloro-8-methylpyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (56.0mg, 0.13mmol) and EtOH (1.0mL) in suspension. The reaction mixture was stirred at room temperature for 1 hour, and then distilled under reduced pressure. Adding Et2O was added to the residue to form a solid. The solid formed is filtered off and the filtrate is,drying under reduced pressure to obtain brown solid compound (1- (7-bromo-2-hydrazino-8-methylpyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester. Unpurified (1- (7-bromo-2-hydrazino-8-methylpyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester was dissolved in trimethyl orthoformate (2.0mL) and stirred at 85 ℃ for 3 hours. The reaction mixture was cooled to room temperature and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give the compound (1- (8-bromo-9-methylpyrido [2, 3-e) as a pale brown solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (25.0mg, 45%).
LC/MS ESI(+)：448(M+1)，450(M+3)
(f) Synthesis of 1- (8-bromo-9-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (8-bromo-9-methylpyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]To a mixture of t-butyl pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (25.0mg, 0.06mmol) and DCM (0.6mL) was stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Adding Et2O is added to the residue to form a solid, the solid formed is filtered and dried under reduced pressure to obtain the compound 1- (8-bromo-9-methylpyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (13.0mg, 68%).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.82(s，1H)，8.63(s，1H)，4.90(m，1H)，4.45-4.35(m，2H)，3.99(m，1H)，3.71(m，1H)，2.90(s，3H)，2.30(s，3H)
Example 66
Synthesis of 8-bromo-9-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 7-bromo-2-chloro-8-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
TEA (72.0. mu.L, 0.51mmol) was added to a mixture of 7-bromo-2, 3-dichloro-8-methylpyrido [2,3-b ] pyrazine (50.0mg, 0.17mmol), N-methylpiperazine (20.0. mu.L, 0.18mmol) and DCM (1.7mL) at 0 ℃ and stirred for 1 h. The reaction mixture was purified by amine silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to yield the compound 7-bromo-2-chloro-8-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine as a yellow solid (50.0mg, 82%).
LC/MS ESI(+)：356(M+1)，358(M+3)，360(M+5)
1H-NMR(300MHz，DMSO-d6)；＝8.96(s，1H)，3.64-3.58(m，4H)，2.69(s，3H)，2.55-2.50(m，4H)，2.25(s，3H)
(b) Synthesis of 8-bromo-9-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Hydrazine monohydrate (22.0. mu.L, 0.70mmol) was added to 7-bromo-2-chloro-8-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (50.0mg, 0.14mmol) and EtOH (1.0 mL). The reaction mixture was stirred at room temperature for 1 hour, and then distilled under reduced pressure. Adding Et2O to the residue to form a solid, filtering the solid formed, and then drying under reduced pressure to obtain 7-bromo-2-hydrazino-8-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b as a yellow solid compound]A pyrazine. Unpurified 7-bromo-2-hydrazino-8-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine was dissolved in trimethyl orthoformate (2.0mL) and then stirred at 85 ℃ for 3 hours. The reaction mixture was then cooled to room temperature and distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). Collecting the product-containing fractions, and evaporating to obtain 8-bromo-9-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazine (24.0mg, 47%).
LC/MS ESI(+)：362(M+1)，364(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.85(s，1H)，8.66(s，1H)，4.60-4.30(m，4H)，2.92(s，3H)，2.54-2.50(m，4H)，2.25(s，3H)
Example 67
Synthesis of 1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of N- (4-chloropyridin-2-yl) pivaloamide
4-Chloropyridin-2-amine (1500.0mg, 11.70mmol) was dissolved in pyridine (6.0mL) and pivaloyl chloride (2.2mL, 17.50mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, poured into water and extracted with EtOAc (100.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc ═ 90:10-80: 20). Fractions containing the product were collected and evaporated to yield the compound N- (4-chloropyridin-2-yl) pivalamide as a white solid (2460.0mg, 99%).
LC/MS ESI(+)：213(M+1)，215(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.12(s，1H)，8.32(d，1H，J＝5.3Hz)，8.16(s，1H)，7.25(d，1H，J＝5.3Hz)，1.23(s，9H)
(b) Synthesis of N- (4, 5-dichloropyridin-2-yl) pivaloamide
N- (4-Chloropyridin-2-yl) pivaloamide (2160.0mg, 10.16mmol) and N-chlorosuccinimide (6781.0mg, 50.78mmol) were dissolved in anhydrous CH3CN (100.0 mL). The mixture was stirred at 70 ℃ for 2 hours and then cooled to room temperature. The reaction mixture was poured into water and extracted with EtOAc (100.0 mL). By 1The organic layer was washed with N NaOH and brine, and anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc ═ 90: 10). The product-containing fractions were collected and evaporated to yield the compound N- (4, 5-dichloropyridin-2-yl) pivaloamide (1990.0mg, 79%) as a cream white solid.
LC/MS ESI(+)：247(M+1)，249(M+3)，251(M+5)
1H-NMR(300MHz，DMSO-d6)；：10.29(s，1H)，8.54(s，1H)，8.34(s，1H)，1.23(s，9H)
(c) Synthesis of 4, 5-dichloro-3-nitropyridine-2-amine
N- (4, 5-dichloropyridin-2-yl) pivaloamide (1990.0mg, 8.05mmol) was added slowly to concentrated H at 10 deg.C2SO4(11.0mL) and dissolved therein, and concentrated HNO was slowly added thereto3Concentrated H2SO4(332.0. mu.L/415.0. mu.L). The reaction mixture was stirred at room temperature for 2.5 hours, poured into ice water, basified with 1N aqueous NaOH (pH 9), and extracted with EtOAc (200.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc ═ 90:10-80: 20). The product-containing fractions were collected and evaporated to give 4, 5-dichloro-3-nitropyridin-2-amine (548.0mg, 32%) as a yellow solid.
LC/MS ESI(+)：208(M+1)，210(M+3)，212(M+5)
(d) Synthesis of 4, 5-dichloropyridine-2, 3-diamine
4, 5-dichloro-3-nitropyridine-2-amine (548.0mg, 2.63mmol), Zn powder (1274.0mg, 19.50mmol) and anhydrous CaCl2(1578.0mg, 14.20mmol) was added to 95% EtOH (20.0mL) and the mixture was stirred at 100 ℃ for 1 h. The reaction mixture was filtered through celite and evaporated under reduced pressure to give 4, 5-dichloropyridine-2, 3-diamine as a brown solid.
LC/MS ESI(+)：178(M+1)，180(M+3)，182(M+5)
(e) Synthesis of 2,3,7, 8-tetrachloropyrido [2,3-b ] pyrazine
Unpurified 4, 5-dichloropyridine-2, 3-diamine was added to diethyl oxalate (10.0 mL). The mixture was stirred at 120 ℃ for 12 hours and then cooled to room temperature. Filtration of Et2O to form a solid, filtering the solid formed, drying under reduced pressure to obtain the compound 7, 8-dichloropyrido [2,3-b ] as a light brown solid]Pyrazine-2, 3(1H,4H) -diones. Unpurified 7, 8-dichloropyrido [2,3-b ]]Pyrazine-2, 3(1H,4H) -diones and POCl3(10.0mL) of the mixture was stirred at 130 ℃ for 48 hours and then cooled to room temperature. The reaction mixture was poured into ice water to form a solid, and the formed solid was filtered and then dried under reduced pressure to obtain 2,3,7, 8-tetrachloropyrido [2,3-b ] as a brown solid compound]Pyrazine (290.0mg, 41%, 3 steps).
1H-NMR(300MHz，DMSO-d6)；：9.35(s，1H)
(f) Synthesis of methyl (1- (2,7, 8-trichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester
2,3,7, 8-tetrachloropyrido [2,3-b ]]Pyrazine (100.0mg, 0.37mmol) and TEA (0.3mL, 1.85mmol) were dissolved in DCM (8.0mL) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (83.0mg, 0.37mmol) diluted with DCM (2.0mL) and TEA (0.3mL, 1.85mmol) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour and poured into saturated NH4Aqueous Cl and then extracted with DCM (30.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and distilling under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 100: 0). The product-containing fractions were collected and evaporated to give methyl (1- (2,7, 8-trichloropyrido [2, 3-b) as a brown solid]Pyrazin-3-yl) azetidin-3-yl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：418(M+1)，420(M+3)
(g) Synthesis of tert-butyl (1- (7, 8-dichloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
Unpurified tert-butyl methyl (1- (2,7, 8-trichloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) carbamate and hydrazine monohydrate (46.0mg, 0.93mmol) were dissolved in EtOH (3.0 mL). The mixture was stirred at room temperature for 20 minutes and distilled under reduced pressure to give tert-butyl (1- (7, 8-dichloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate.
LC/MS ESI(+)：414(M+1)，416(M+3)
(h) Synthesis of tert-butyl (1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Unpurified (1- (7, 8-dichloro-2-hydrazinopyrido [2, 3-b))]A mixture of t-butyl pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure to obtain (1- (8, 9-dichloropyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
LC/MS ESI(+)：424(M+1)，426(M+3)
(i) Synthesis of 1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
Unpurified tert-butyl (1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate was dissolved in DCM (2.0mL) and TFA (0.5mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour and distilled under reduced pressure. The residue was neutralized with TEA and purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to give 1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine as a cream solid (3.5mg, 3%, 4 steps).
LC/MS ESI(+)：324(M+1)，326(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.30(s，1H)，8.68(s，1H)，4.94(m，1H)，4.46(m，2H)，4.03(m，1H)，3.73(m，1H)，2.30(s，3H)
Example 68
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride
A mixture of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (200.0mg, 0.60mmol) and 4N HCl in dioxane (10.0mL) was stirred at room temperature for 12 hours. The solid formed was filtered and then dried to give the compound 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride as a white solid (200.0mg, 90%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：10.0(s，1H)，9.69(bs，2H)，9.00(d，1H，J＝2.2Hz)，8.66(d，1H，J＝2.2Hz)，5.20～4.80(m，2H)，4.70～4.40(m，2H)，4.20(m，1H)，2.64(m，3H)
Example 69
Synthesis of 1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride
1- (8-bromopyrido [2,3-e ]]Tetrazolo [1,5-a ] s]A suspension of pyrazin-4-yl) -N-methylazetidin-3-amine (190.0mg, 0.57mmol) and 4N HCl in dioxane (5.0mL) was stirred at room temperature for 8 hours. Et was added thereto2O to form a solid, filtering the solid formed, drying under reduced pressure to obtain a yellow solidified productCompound 1- (8-bromopyrido [2, 3-e)]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride (210.0mg, 99%).
LC/MS ESI(+)：335(M+1)，337(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.77(bs，2H)，9.00(d，1H，J＝2.4Hz)，8.88(d，1H，J＝2.4Hz)，5.10-4.90(m，2H)，4.75-4.45(m，2H)，4.31(m，1H)，2.66(s，3H)
Example 70
Synthesis of 8-chloro-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ] pyrazine (100.0mg, 0.43mmol) and TEA (0.6mL, 4.26mmol) were dissolved in DCM (3.0mL) and 2-methyloctahydropyrrolo [3,4-c ] pyrrole (59.1mg, 0.47mmol) diluted with DCM (1.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound 2, 7-dichloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (72.0mg, 52%) as a yellow solid.
LC/MS ESI(+)：324(M+1)，326(M+3)
(b) Synthesis of 7-chloro-2-hydrazino-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine
2, 7-dichloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (72.0mg, 0.22mmol) and hydrazine monohydrate (32.0. mu.L, 0.67mmol) were dissolved in EtOH (3.0 mL). The mixture was stirred at room temperature for 12 hours to form a solid. The solid formed was filtered to give the compound 7-chloro-2-hydrazino-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (71.0mg, 100%) as a yellow solid.
LC/MS ESI(+)：320(M+1)，322(M+3)
(c) Synthesis of 8-chloro-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 7-chloro-2-hydrazino-3- (5-methyl hexahydro pyrrolo [3, 4-c)]Pyrrol-2 (1H) -yl) pyrido [2,3-b]A mixture of pyrazine (71.0mg, 0.22mmol) and trimethyl orthoformate (3.0mL) was stirred at 80 ℃ for 3 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain 8-chloro-4- (5-methylhexahydropyrrolo [3, 4-c) as yellow solid]Pyrrol-2 (1H) -yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (20.0mg, 27%).
LC/MS ESI(+)：330(M+1)，332(M+3)
1H-NMR(300MHz，CDCl3)；：9.14(s，1H)，8.58(d，1H，J＝2.4Hz)，8.01(d，1H，J＝2.4Hz)，4.70-4.58(m，2H)，4.23-4.02(m，2H)，3.14-3.03(m，2H)，2.71-2.65(m，4H)，2.34(s，3H)
Example 71
Synthesis of 8-bromo-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 7-bromo-2-chloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine
7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (100.0mg, 0.36mmol) and TEA (0.5mL, 3.58mmol) were dissolved in DCM (3.0mL) and 2-methyloctahydropyrrolo [3,4-c ] pyrrole (49.6mg, 0.39mmol) diluted with DCM (1.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 12 hours, then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound 7-bromo-2-chloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (63.0mg, 48%) as a yellow solid.
LC/MS ESI(+)：368(M+1)，370(M+3)
(b) Synthesis of 7-bromo-2-hydrazino-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine
7-bromo-2-chloro-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (63.0mg, 0.17mmol) and hydrazine monohydrate (25.0. mu.L, 0.51mmol) were dissolved in EtOH (3.0 mL). The mixture was stirred at room temperature for 12 hours to form a solid. The solid formed was filtered to give the compound 7-bromo-2-hydrazino-3- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-b ] pyrazine (62.0mg, 100%) as a yellow solid.
LC/MS ESI(+)：364(M+1)，366(M+3)
(c) Synthesis of 8-bromo-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 7-bromo-2-hydrazino-3- (5-methyl hexahydro-pyrrolo [3, 4-c)]Pyrrol-2 (1H) -yl) pyrido [2,3-b]A mixture of pyrazine (62.0mg, 0.17mmol) and trimethyl orthoformate (3.0mL) was stirred at 90 deg.C for 6 hours, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure. The residue was purified by amine silica gel column chromatography (EtOAc: n-Hex: MeOH ═ 4:4: 1). Collecting the product-containing fractions, and evaporating to obtain 8-bromo-4- (5-methylhexahydropyrrolo [3, 4-c) as yellow solid]Pyrrol-2 (1H) -yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (11.0mg, 16%).
LC/MS ESI(+)：374(M+1)，376(M+3)
1H-NMR(300MHz，CDCl3)；：9.17(s，1H)，8.64(d，1H，J＝2.1Hz)，8.14(d，1H，J＝2.1Hz)，4.73-4.55(m，2H)，4.22-3.98(m，2H)，3.12-3.02(m，2H)，2.63(m，4H)，2.32(s，3H)
Example 72
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) carbamate
(3-Methylazetidin-3-yl) carbamic acid tert-butyl ester hydrochloride (43.0mg, 0.23mmol) and TEA (0.8mL, 0.54mmol) were added to a mixture of 7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (50.0mg, 0.18mmol) and DCM (1.8mL) at 0 deg.C and stirred for 1 hour. The reaction mixture was concentrated under reduced pressure to give a yellow solid compound, tert-butyl 1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) carbamate.
LC/MS ESI(+)：428(M+1)，430(M+3)
(b) Synthesis of (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) carbamic acid tert-butyl ester
Hydrazine monohydrate (28.0. mu.L, 0.90mmol) was added to a suspension of unpurified tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) carbamate and EtOH (1.0 mL). The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure to give the compound tert-butyl 1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) carbamate as a yellow solid.
LC/MS ESI(+)：424(M+1)，426(M+3)
(c) Synthesis of tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-yl) carbamate
Unpurified (1- (7-bromo-2-hydrazinopyrido [2, 3-b))]A suspension of pyrazin-3-yl) -3-methylazetidin-3-yl) carbamic acid tert-butyl ester and trimethyl orthoformate (3.0mL) was stirred at 90 ℃ for 3 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, and reducing the pressure to obtain the light brown solid compound (1- (8-bromopyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -3-methylazetidin-3-yl) carbamic acid tert-butyl ester (25.0mg, 32%, 3 steps).
LC/MS ESI(+)：434(M+1)，436(M+3)
(d) Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-amine
Tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-yl) carbamate (25.0mg, 0.06mmol) was dissolved in DCM (0.6mL), to which was added TFA (0.4 mL). The reaction mixture was stirred at room temperature for 1 hour, then purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to yield 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-amine as a brown solid (9.0mg, 47%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.90(d，1H，J＝2.4Hz)，8.59(d，1H，J＝2.4Hz)，4.58(m，1H)，4.47(m，1H)，4.15-3.95(m，2H)，2.40(bs，2H)，1.43(s，3H)
Example 73
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, 3-dimethylazetidin-3-amine
(a) Synthesis of benzyl 3- ((tert-butoxycarbonyl) amino) -3-methylazetidine-1-carboxylate
Tert-butyl (3-methylazetidin-3-yl) carbamate hydrochloride (400.0mg, 1.80mmol) was dissolved in DCM (9.0mL) and benzyl chloroformate (0.8mL, 5.39mmol) and TEA (1.3mL, 9.00mmol) were added thereto at 0 ℃. The reaction mixture was reacted at room temperature for 12 hours, and then purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and evaporated to give the compound benzyl 3- ((tert-butoxycarbonyl) amino) -3-methylazetidine-1-carboxylate (680.0mg, 99%) as a white solid.
LC/MS ESI(+)：321(M+1)
1H-NMR(300MHz，CDCl3)；：7.38-7.30(m，5H)，5.10(s，2H)，4.76(bs，1H)，4.20-4.10(m，2H)，3.90-3.75(m，2H)，1.53(s，3H)，1.44(m，9H)
(b) Synthesis of benzyl 3- ((tert-butoxycarbonyl) (methyl) amino) -3-methylazetidine-1-carboxylate
Benzyl 3- ((tert-butoxycarbonyl) amino) -3-methylazetidine-1-carboxylate (676.0mg, 2.11mmol) was dissolved in THF (21.0mL), to which was added NaH (110.0mg, 2.74 mmol). The reaction mixture was stirred for 10 minutes, then MeI (0.2mL, 3.17mmol) was added thereto, stirred at room temperature for 12 hours, and then reacted at 50 ℃ for 12 hours. The reaction mixture was cooled to room temperature and purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 7). The product-containing fractions were collected and evaporated to give benzyl 3- ((tert-butoxycarbonyl) (methyl) amino) -3-methylazetidine-1-carboxylate (180.0mg, 26%) as a viscous colorless liquid.
LC/MS ESI(+)：335(M+1)
1H-NMR(300MHz，CDCl3)；：7.40-7.30(m，5H)，5.11(s，2H)，4.12-4.05(m，2H)，3.75-3.60(m，2H)，2.69(s，3H)，1.48(s，3H)，1.45(m，9H)
(c) Synthesis of methyl (3-methylazetidin-3-yl) carbamic acid tert-butyl ester
Benzyl 3- ((tert-butoxycarbonyl) (methyl) amino) -3-methylazetidine-1-carboxylate (180.0mg, 0.54mmol) was dissolved in MeOH (2.0mL) to which was added 10% Pd/C (18.0 mg). The flask was replaced with hydrogen and stirred for 12 hours. The reaction mixture was filtered through celite and distilled under reduced pressure to give tert-butyl methyl (3-methylazetidin-3-yl) carbamate (50.0mg, 46%) as a viscous colorless liquid.
1H-NMR(300MHz，DMSO-d6)；：3.85-3.75(m，2H)，3.23-3.15(m，2H)，2.64(s，3H)，1.52(s，3H)，1.44(m，9H)
(d) Synthesis of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate
TEA (90.0. mu.L, 0.65mmol) was added to a mixture of tert-butyl methyl (3-methylazetidin-3-yl) carbamate (47.0mg, 0.24mmol), 7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (60.0mg, 0.22mmol) and DCM (2.2mL) at 0 ℃ and stirred for 1 h. The reaction mixture was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate as a yellow solid (10.0mg, 11%).
LC/MS ESI(+)：442(M+1)，444(M+3)
(e) Synthesis of tert-butyl (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate
Tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate (10.0mg, 0.02mmol) was dissolved in EtOH (1.0mL), to which was added hydrazine monohydrate (3.5 μ L, 0.11 mmol). The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure to give the compound tert-butyl (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate as a yellow solid.
LC/MS ESI(+)：438(M+1)，440(M+3)
(f) Synthesis of tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-yl) (methyl) carbamate
Unpurified tert-butyl (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -3-methylazetidin-3-yl) (methyl) carbamate was dissolved in trimethyl orthoformate (1.0mL) and stirred at 80 ℃ for 1 hour. The reaction mixture was then cooled to room temperature and distilled under reduced pressure to give the atypical yellow compound tert-butyl 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-yl) (methyl) carbamate.
LC/MS ESI(+)：448(M+1)，450(M+3)
(g) Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, 3-dimethylazetidin-3-amine
Unpurified tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-yl) (methyl) carbamate was dissolved in 4N HCl in dioxane, and the reaction mixture was stirred at room temperature for 1 hour and distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated to give the compound 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, 3-dimethylazetidin-3-amine as a cream-white solid (3.0mg, 38%, 3 steps).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.90(d，1H，J＝2.4Hz)，8.59(d，1H，J＝2.4Hz)，4.60-4.40(m，2H)，4.18-3.96(m，2H)，2.27(s，3H)，1.41(s，3H)
Example 74
Synthesis of 8-bromo-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 7-bromo-2-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-b ] pyrazine
7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (206.0mg, 0.74mmol) and octahydropyrrolo [1,2-a ] pyrazine (102.0mg,
0.81mmol) to yield the compound 7-bromo-2-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-b ] pyrazine (180.0mg, 66%) as a yellow solid.
LC/MS ESI(+)：370(M+1)，372(M+3)
1H-NMR(300MHz，CDCl3)；：8.93(d，1H，J＝2.4Hz)，8.32(d，1H，J＝2.4Hz)，4.46(m，2H)，3.15(m，3H)，2.92(m，1H)，2.48(m，1H)，2.22(m，2H)，1.89(m，3H)，1.57(m，1H)
(b) Synthesis of 8-bromo-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 7-bromo-2-chloro-3- (hexahydropyrrolo [1,2-a ]]Pyrazin-2 (1H) -yl) pyrido [2,3-b]Pyrazine (100.0mg, 0.36mmol) and hydrazine monohydrate (0.028mL, 0.90mmol) were dissolved in EtOH (8.0mL) and the mixture was stirred at 40 ℃ for 12 h. Et was added thereto2O/EtOH to form a solid, filtering the solid formed, drying under reduced pressure to give the yellow compound 7-bromo-3- (hexahydropyrrolo [1, 2-a)]Pyrazin-2 (1H) -yl) -2-hydrazinopyrido [2,3-b]A pyrazine. Unpurified 7-bromo-3- (hexahydropyrrolo [1,2-a ] mixture]Pyrazin-2 (1H) -yl) -2-hydrazinopyrido [2,3-b]A mixture of pyrazine (60.0mg, 0.17mmol) and trimethyl orthoformate (1.0mL) was stirred at 80 deg.C for 1.5 hours, then cooled to room temperature. Et was added thereto2O to form a solid, filtering the solid formed, drying under reduced pressure to obtain the brown solid compound 8-bromo-4- (hexahydropyrrolo [1,2-a ]]Pyrazin-2 (1H) -yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (35.0mg, 26%, 2 steps).
LC/MS ESI(+)：374(M+1)，376(M+3)
1H-NMR(300MHz，CDCl3)；：9.16(s，1H)，8.70(d，1H，J＝2.1Hz)，8.17(d，1H，J＝2.1Hz)，6.40(m，1H)，5.52(m，1H)，3.57(m，1H)，3.20(m，2H)，2.92(m，1H)，2.44(m，1H)，2.20(m，2H)，1.79(m，3H)，1.55(m，1H)
Example 75
Synthesis of 4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -7-iodopyrido [2,3-b ] pyrazine
2, 3-dichloro-7-iodopyrido [2,3-b ] pyrazine (50.0mg, 0.15mmol) and TEA (213.0. mu.L, 1.53mmol) were dissolved in DCM (2.0mL) and octahydropyrrolo [1,2-a ] pyrazine (19.0mg, 0.15mmol) was added thereto at 0 ℃. The reaction mixture was stirred at 0 ℃ for 1 hour, then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 3: 97). The product-containing fractions were collected and evaporated to yield the compound 2-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -7-iodopyrido [2,3-b ] pyrazine (38.0mg, 58%) as a yellow solid.
LC/MS ESI(+)：416(M+1)，418(M+3)
(b) Synthesis of 3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -2-hydrazino-7-iodopyrido [2,3-b ] pyrazine
2-chloro-3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -7-iodopyrido [2,3-b ] pyrazine (38.0mg, 0.09mmol) and hydrazine monohydrate (6.7. mu.L, 0.14mmol) were dissolved in EtOH (1.0 mL). The mixture was stirred at room temperature for 3 hours to form a solid, and the solid formed was filtered to give the compound 3- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -2-hydrazino-7-iodopyrido [2,3-b ] pyrazine (37.5mg, 100%) as a yellow solid.
LC/MS ESI(+)：412(M+1)
(c) Synthesis of 4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 3- (hexahydropyrrolo [1,2-a ]]Pyrazin-2 (1H) -yl) -2-hydrazino-7-iodopyrido [2,3-b]A mixture of pyrazine (39.4mg, 0.10mmol) and trimethyl orthoformate (2.0mL) was stirred at 80 ℃ for 3 hours and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain 4- (hexahydropyrrolo [1,2-a ] compound as yellow solid]Pyrazin-2 (1H) -yl) -8-iodopyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (9.0mg, 26%).
LC/MS ESI(+)：422(M+1)
1H-NMR(300MHz，DMSO-d6)；：10.01(s，1H)，9.01(d，1H，J＝2.1Hz)，8.71(d，1H，J＝2.1Hz)，6.18-5.25(m，1H)，3.42-3.35(m，1H)，3.28-3.24(m，1H)，3.20-3.15(m，1H)，3.08-2.87(m，2H)，2.25(m，1H)，2.15-2.03(m，2H)，1.88(m，1H)，1.81-1.68(m，2H)，1.45(m，1H)
Example 76
Synthesis of 8-chloro-4- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ] pyrazine (70.0mg, 0.30mmol) and 1-cyclopropylpiperazine (59.7mg, 0.30mmol) were reacted in the same manner as in example 2(c) to give 2, 7-dichloro-3- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (59.5mg, 61%) as a yellow solid.
LC/MS ESI(+)：324(M+1)，326(M+3)
1H-NMR(300MHz，CDCl3)；：8.93(d，1H，J＝2.7Hz)，8.50(d，1H，J＝2.7Hz)，3.59(m，4H)，2.74(m，4H)，1.71(m，1H)，0.45(m，2H)，0.39(m，2H)
(b) Synthesis of 7-chloro-3- (4-cyclopropylpiperazin-1-yl) -2-hydrazinopyrido [2,3-b ] pyrazine
2, 7-dichloro-3- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (56.7mg, 0.17mmol) and hydrazine monohydrate (14.0 μ L, 0.43mmol) were reacted in the same manner as in example 2(d) to give 7-chloro-3- (4-cyclopropylpiperazin-1-yl) -2-hydrazinopyrido [2,3-b ] pyrazine (50.0mg, 100%) as an orange solid.
LC/MS ESI(+)：320(M+1)，322(M+3)
(c) Synthesis of 8-chloro-4- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
7-chloro-3- (4-cyclopropylpiperazin-1-yl) -2-hydrazinopyrido [2,3-b ] pyrazine (64.0mg, 0.14mmol) and trimethyl orthoformate (2.0mL) were reacted at 85 ℃ for 2 hours in the same manner as in example 2(e) to give 8-chloro-4- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream solid (40.6mg, 62%).
LC/MS ESI(+)：330(M+1)，332(M+3)
1H-NMR(300MHz，CDCl3)；：10.01(s，1H)，8.85(d，1H，J＝2.7Hz)，8.57(d，1H，J＝2.7Hz)，4.80-3.93(m，4H)，2.73(m，4H)，1.68(m，1H)，0.46(m，2H)，0.40(m，2H)
Example 77
Synthesis of 4- ((1S,4S) -2, 5-diazabicyclo [2.2.1] hept-2-yl) -8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl (1S,4S) -5- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) -2, 5-diazabicyclo [2.2.1] hepta-2-carboxylate
TEA (0.2mL, 1.28mmol) was added to a mixture of tert-butyl 2,3, 7-trichloropyrido [2,3-b ] pyrazine (100.0mg, 0.43mmol), (1S,4S) -2, 5-diazabicyclo [2.2.1] hepta-2-carboxylate (89.0mg, 0.45mmol) in DCM (4.3mL) at 0 deg.C, then stirred for 3 hours. The reaction mixture was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to give the compound tert-butyl (1S,4S) -5- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) -2, 5-diazabicyclo [2.2.1] hepta-2-carboxylate (120.0mg, 71%) as a yellow solid.
LC/MS ESI(+)：396(M+1)，398(M+3)
1H-NMR(300MHz，CDCl3)；：8.79(d，1H，J＝2.7Hz)，8.11(d，1H，J＝2.7Hz)，5.30(m，1H)，4.78-4.55(m，1H)，4.20(m，1H)，3.95-3.63(m，2H)，3.53(m，1H)，1.99(m，2H)，1.56(m，9H)
(b) Synthesis of tert-butyl (1S,4S) -5- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -2, 5-diazabicyclo [2.2.1] heptan-2-carboxylate
Hydrazine monohydrate (48.0. mu.L, 1.51mmol) was added to (1S,4S) -5- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) -2, 5-diazabicyclo [2.2.1]Tert-butyl hepta-2-carboxylate (120.0mg, 0.30mmol) and EtOH (1.5 mL). The reaction mixture was stirred at room temperature for 1 hour. Et was then added thereto2O to form a solid, filtering the formed solid, and drying under reduced pressure. Trimethyl orthoformate (3.0mL) was added to the solid formed. The mixture was stirred at 85 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid, filtering the formed solid, and then drying under reduced pressure to obtain (1S,4S) -5- (8-chloropyrido [2, 3-e) as a yellow solid compound][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -2, 5-diazabicyclo [2.2.1]Tert-butyl hepta-2-carboxylate (91.0mg, 83%).
LC/MS ESI(+)：402(M+1)，404(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.98(m，1H)，8.83(m，1H)，8.56(m，1H)，6.27(s，0.7H)，5.27(s，0.3H)，4.57(m，1H)，4.24(m，0.7H)，3.80(m，1H)，3.50(m，1H)，3.40(m，1.3H)，2.20-1.98(m，2H)，1.38-1.49(m，9H)
(c) Synthesis of 4- ((1S,4S) -2, 5-diazabicyclo [2.2.1] hept-2-yl) -8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
TFA (0.8mL) was added to (1S,4S) -5- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -2, 5-diazabicyclo [2.2.1]Tert-butyl hepta-2-carboxylate (90.0mg, 0.22mmol) and DCM (1.2mL), then stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated. Adding Et2O is added to the residue to form a solid, the solid formed is filtered and dried under reduced pressure to give 4- ((1S,4S) -2, 5-diazabicyclo [2.2.1] compound as a milky white solid]Hept-2-yl) -8-chloropyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (58.0mg, 85%).
LC/MS ESI(+)：302(M+1)，304(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.96(m，1H)，8.79(m，1H)，8.52(s，1H)，6.19(s，0.7H)，5.17(s，0.3H)，4.16(s，0.7H)，3.78(s，1H)，3.70(m，1.3H)，3.12-2.80(m，2H)，1.95-1.70(m，2H)
Example 78
Synthesis of 8-chloro-4- ((1S,4S) -5-methyl-2, 5-diazabicyclo [2.2.1] hept-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
37% Formaldehyde (28.0. mu.L, 0.38mmol) and NaBH were added4(14.0mg, 0.38mmol) was added to 4- ((1S,4S) -2, 5-diazabicyclo [2.2.1]Hept-2-yl) -8-chloropyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (38.0mg, 0.13mmol) and MeOH (1.3mL) were suspended. The reaction mixture was stirred at room temperature for 1 hour, and then distilled under reduced pressure. EtOAc and water were added to the residue, extracted with EtOAc,with anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected, evaporated and then DCM/Et was added thereto2O to form a solid. The solid formed was filtered and then dried under reduced pressure to give 8-chloro-4- ((1S,4S) -5-methyl-2, 5-diazabicyclo [2.2.1] compound as a milky white solid]Hept-2-yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (20.0mg, 50%).
LC/MS ESI(+)：316(M+1)，318(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.96(m，1H)，8.79(m，1H)，8.53(m，1H)，6.13(s，0.7H)，5.13(s，0.3H)，4.40(m，0.3H)，4.04(m，0.3H)，3.88(m，0.7H)，3.60(m，1.7H)，2.97(m，1H)，2.64(m，1H)，2.36(s，3H)，2.08-1.80(m，2H)
Example 79
Synthesis of 8-chloro-4- (1, 4-diazacycloheptan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 4- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) -1, 4-diazepan-1-carboxylate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (70.0mg, 0.30mmol) was dissolved in DCM (3.0mL) and 1, 4-diazepan-1-carboxylic acid tert-butyl ester (60.0. mu.L, 0.30mmol) and TEA (135.0. mu.L, 0.90mmol) were added thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, then diluted with DCM. The organic layer was washed with water and brine, and anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (n-Hex: EtOAc ═ 3: 1). Collecting the product-containing fractions, and concentrating to obtain 4- (2, 7-dichloropyrido [2,3-b ] yellow liquid]Pyrazin-3-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester (85.0mg, 71%).
LC/MS ESI(+)：398(M+1)，400(M+3)
1H-NMR(300MHz，CDCl3)；：8.82(d，1H，J＝2.67Hz)，8.12(d，1H，J＝2.67Hz)，4.15-3.92(m，4H)，3.78-3.70(m，2H)，3.57-3.38(m，2H)，2.18-2.06(m，2H)，1.41(s，9H)
(b) Synthesis of 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester
Hydrazine monohydrate (6.5. mu.L, 0.60mmol) was added to 4- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester (82.0mg, 0.20mmol) and EtOH (2.0mL), then stirred at room temperature for 3 hours. With Et2The reaction mixture was diluted with O and then stirred at room temperature for 1 hour. The solid formed was filtered and dried under reduced pressure to give 4- (7-chloro-2-hydrazinopyrido [2,3-b ] as a yellow solid]Pyrazin-3-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：394(M+1)，396(M+3)
(c) Synthesis of 8-chloro-4- (1, 4-diazacycloheptan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
A mixture of unpurified tert-butyl 4- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) -1, 4-diazepan-1-carboxylate and trimethyl orthoformate (1.1mL) was stirred at 80 ℃ for 2 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure. Unpurified tert-butyl 4- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -1, 4-diazepan-1-carboxylate was dissolved in DCM (1.0mL), to which TFA (0.5mL) was slowly added at room temperature, followed by stirring at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 10: 1). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-4- (1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a yellow solid (28.0mg, 35%, 3 steps).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，CDCl3)；：9.14(s，1H)，8.60(d，1H，J＝2.3Hz)，8.01(d，1H，J＝2.3Hz)，4.84-4.72(m，2H)，4.28-4.17(m，2H)，3.21(t，2H，J＝5.7Hz)，2.94(t，2H，J＝5.7Hz)，2.12-2.02(m，2H)
Example 80
Synthesis of 8-chloro-4- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 2, 7-dichloro-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b ] pyrazine
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (50.0mg, 0.21mmol) was dissolved in DCM (2.0mL) and 1-methyl-1, 4-diazepane (26.1. mu.L, 0.21mmol) and TEA (94.7. mu.L, 0.63mmol) were added thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, then diluted with DCM. The organic layer was washed with water and brine, and anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 10: 1). Collecting the product-containing fractions, and concentrating to obtain brown solid compound 2, 7-dichloro-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b]Pyrazine (42.0mg, 64%).
LC/MS ESI(+)：312(M+1)，314(M+3)
1H-NMR(300MHz，CDCl3)；：8.80(d，1H，J＝2.6Hz)，8.11(d，1H，J＝2.6Hz)，4.09-4.03(m，2H)，4.01-3.95(m，2H)，3.02-2.93(m，2H)，2.78-2.69(m，2H)，2.46(s，3H)，2.25-2.13(m，2H)
(b) Synthesis of 7-chloro-2-hydrazino-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b ] pyrazine
Hydrazine monohydrate (3.9 μ L, 0.36mmol) was added to 2, 7-dichloro-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b]Pyrazine (38.5mg, 0.12mmol) and EtOH (1.2mL)Then, it was stirred at room temperature for 2 hours. With Et2The reaction mixture was diluted with O and then stirred at room temperature for 1 hour. The solid formed was filtered and dried under reduced pressure to give the unpurified yellow solid compound 7-chloro-2-hydrazino-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b]A pyrazine.
LC/MS ESI(+)：308(M+1)，310(M+3)
(c) Synthesis of 8-chloro-4- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
A mixture of unpurified 7-chloro-2-hydrazino-3- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-b ] pyrazine and trimethyl orthoformate (0.7mL) was stirred at 80 ℃ for 2 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure and then purified by amine silica gel column chromatography (DCM: MeOH ═ 20: 1). The product-containing fractions were collected and concentrated to give 8-chloro-4- (4-methyl-1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a yellow solid (16.0mg, 42%, 2 steps).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，CDCl3)；：9.14(s，1H)，8.59(d，1H，J＝2.3Hz)，8.01(d，1H，J＝2.3Hz)，4.87-4.74(m，2H)，4.18-4.14(m，2H)，2.92-2.86(m，2H)，2.67-2.60(m，2H)，2.39(s，3H)，2.21-2.09(m，2H)
Example 81
Synthesis of (R) -1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine
(a) Synthesis of (R) - (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester
7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (70.0mg, 0.25mmol) and TEA (175.0. mu.L, 1.26mmol) were dissolved in DCM (1.0mL) and tert-butyl (R) -methyl (pyrrolidin-3-yl) carbamate (55.2mg, 0.28mmol) dissolved in DCM (0.5mL) was added thereto at 0 ℃. The reaction mixture was stirred at 0 ℃ for 12 hours, then concentrated to give tert-butyl (R) - (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate (111.1mg, 100%).
LC/MS ESI(+)：442(M+1)，444(M+3)
(b) Synthesis of (R) - (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester
Tert-butyl (R) - (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate (111.1mg, 0.25mmol) and hydrazine monohydrate (25.0. mu.L, 0.50mmol) were dissolved in EtOH (1.5 mL). The mixture was stirred at room temperature for 3 hours to form a solid, and the solid formed was filtered to give the compound (R) - (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester as a yellow solid (110.0mg, 100%).
LC/MS ESI(+):438(M+1)，440(M+3)
(c) Synthesis of tert-butyl (R) - (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (R) - (1- (7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) pyrrolidin-3-yl) (methyl) carbamate (110.0mg, 0.25mmol) and trimethyl orthoformate (1.0mL) was stirred at 80 ℃ for 2 hours, then cooled to room temperature. The reaction mixture was concentrated and then purified first by silica gel column chromatography (MeOH: DCM ═ 5:95) and second by amine silica gel column chromatography (EtOAc: n-Hex ═ 50: 50). Fractions containing the product were collected and evaporated to yield the compound (R) - (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamic acid tert-butyl ester as a yellow solid (40.0mg, 36%).
LC/MS ESI(+)：448(M+1)，450(M+3)
(d) Synthesis of (R) -1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine
Tert-butyl (R) - (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-yl) (methyl) carbamate (40.0mg, 0.09mmol) was dissolved in DCM (1.0mL), to which TFA (0.4mL) was added. The reaction mixture was stirred at room temperature for 1 hour, then purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound (R) -1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine (20.7mg, 67%) as a yellow solid.
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.98(s，1H)，8.89(d，1H，J＝2.1Hz)，8.57(d，1H，J＝2.1Hz)，4.36-4.20(m，2H)，3.81-3.61(m，2H)，3.30(m，1H)，2.32(s，3H)，2.11-1.86(m，3H)
Example 82
Synthesis of 8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of tert-butyl 6- (2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate
2,3, 7-trichloropyrido [2,3-b ]]Pyrazine (150.0mg, 0.64mmol) was dissolved in DCM (6.0 mL). To which octahydro-1H-pyrrolo [3,4-b ] is added at 0 deg.C]Pyridine-1-carboxylic acid tert-butyl ester (151.0mg, 0.67mmol) and TEA (288.0. mu.L, 1.92 mmol). The reaction mixture was stirred at room temperature for 2 hours, then diluted with DCM, the organic layer was washed with water and brine, anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (n-Hex: EtOAc ═ 3: 1). Collecting the product-containing fractions, and concentrating to obtain milky white solid compound 6- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b]Pyridine-1-carboxylic acid tert-butyl ester (173).0mg，61％)。
LC/MS ESI(+)：424(M+1)，426(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.81(d，1H，J＝2.7Hz)，8.38(d，1H，J＝2.7Hz)，4.69(m，1H)，4.00-3.80(m，4H)，3.70(m，1H)，2.85(m，1H)，2.26(m，1H)，1.77-1.61(m，2H)，1.42(s，9H)，1.38-1.26(m，2H)
(b) Synthesis of tert-butyl 6- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate
Hydrazine monohydrate (37.7. mu.L, 1.20mmol) was added to 6- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b]Pyridine-1-carboxylic acid tert-butyl ester (170.0mg, 0.40mmol) and EtOH (4.0mL), followed by stirring at room temperature for 3 hours. With Et2The reaction mixture was diluted with O and stirred at room temperature for 1 hour to form a solid. The solid formed was filtered and then dried under reduced pressure to give 6- (7-chloro-2-hydrazinopyrido [2,3-b ] compound as a yellow solid]Pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b]Pyridine-1-carboxylic acid tert-butyl ester.
LC/MS ESI(+)：420(M+1)，422(M+3)
(c) Synthesis of tert-butyl 6- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate
A mixture of unpurified tert-butyl 6- (7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate and trimethyl orthoformate (4.0mL) was stirred at 80 ℃ for 2 hours, then cooled to room temperature. The reaction mixture was concentrated under reduced pressure and purified by amine silica gel column chromatography (n-Hex: EtOAc ═ 3: 1). The product-containing fractions were collected and concentrated to give the compound tert-butyl 6- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate (79.0mg, 46%, 2 steps) as a yellow solid.
LC/MS ESI(+)：430(M+1)，432(M+3)
1H-NMR(300MHz，CDCl3)；：9.16(s，1H)，8.59(d，1H，J＝2.3Hz)，8.03(d，1H，J＝2.3Hz)，4.91(m，1H)，4.75(m，1H)，4.35-4.00(m，3H)，3.94-3.71(m，1H)，2.81(m，1H)，2.37(m，1H)，1.91-1.68(m，2H)，1.62-1.31(m，11H)
(d) Synthesis of 8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Tert-butyl 6- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) octahydro-1H-pyrrolo [3,4-b ] pyridine-1-carboxylate (76.0mg, 0.17mmol) was dissolved in DCM (2.0mL) and slowly TFA (126.0. mu.L, 1.70mmol) was added thereto at room temperature. The mixture was stirred at room temperature for 8 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 10: 1). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a cream-white solid (51.0mg, 91%).
LC/MS ESI(+)：330(M+1)，332(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.77(d，1H，J＝2.7Hz)，8.50(d，1H，J＝2.7Hz)，4.53-4.41(m，1H)，4.27-4.10(m，1H)，3.84-3.64(m，2H)，3.42-3.31(m，1H)，2.89-2.79(m，1H)，2.58-2.51(m，1H)，2.44-2.26(m，2H)，1.84-1.35(m，4H)
Example 83
Synthesis of 8-chloro-4- (1-methylhexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Reacting 8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ]]Pyridin-6 (2H) -yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (35.0mg, 0.10mmol) was dissolved in MeOH (0.7mL), to which 37% formaldehyde (40.5mg, 0.50mmol) dissolved in MeOH (0.3mL) was added at 0 deg.C, and then to whichAdding NaBH4(18.9mg, 0.50 mmol). The mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with EtOAc. The organic layer was then washed with water and brine, and anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 20: 1). Collecting the product-containing fractions, and concentrating to obtain brown solid compound 8-chloro-4- (1-methylhexahydro-1H-pyrrolo [3, 4-b)]Pyridin-6 (2H) -yl) pyrido [2,3-e][1,2,4]Triazolo [4,3-a]Pyrazine (7.4mg, 22%).
LC/MS ESI(+)：344(M+1)，346(M+3)
1H-NMR(300MHz，CDCl3)；：9.15(s，1H)，8.58(d，1H，J＝2.3Hz)，8.02(d，1H，J＝2.3Hz)，5.06(m，0.4H)，4.69-4.60(m，0.6H)，4.51-4.39(m，1H)，4.19(m，0.4H)，4.03(m，1H)，3.80(m，0.6H)，2.89-2.75(m，2H)，2.67-2.47(m，1H)，2.34(s，3H)，2.18(m，1H)，1.95-1.55(m，4H)
Example 84
Synthesis of 1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine
Reacting NaBH4(29.0mg, 0.77mmol) was added to 1- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (74.0mg, 0.26mmol), 37% formaldehyde (29.0. mu.L, 0.38mmol) and MeOH (2.5 mL). The reaction mixture was stirred at room temperature for 5 hours, to which EtOAc and water were added and extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 100). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-chloropyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (12.0mg, 16%).
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.81(d，1H，J＝2.4Hz)，8.54(d，1H，J＝2.4Hz)，4.81(m，1H)，4.53(m，1H)，4.34.(m，1H)，4.11(m，1H)，3.35(m，1H)，2.18(s，6H)
Example 85
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine
Reacting NaBH4(11.0mg, 0.29mmol) was added to 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (32.0mg, 0.10mmol), 37% formaldehyde (11.0. mu.L, 0.14mmol) and MeOH (1.0 mL). The reaction mixture was stirred at room temperature for 5 hours, then EtOAc and water were added thereto and extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 100). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (3.0mg, 9%).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.95(s，1H)，8.90(d，1H，J＝2.4Hz)，8.60(d，1H，J＝2.4Hz)，4.81(m，1H)，4.51(m，1H)，4.31(m，1H)，4.10(m，1H)，3.40(m，1H)，2.18(s，6H)
Example 86
Synthesis of (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid
Purification by amine silica gel column chromatography (DCM: MeOH ═ 90:10) obtained from tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (13000.0mg, 30.3mmol) of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine in the same manner as in examples 3(e) - (f). The product-containing fractions were collected and evaporated to give the compound (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid as a cream-white solid (56.0mg, 0.5%).
LC/MS ESI(+)：378(M+1)，380(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.97(s，1H)，9.03(bs，1H)，8.95(d，1H，J＝2.2Hz)，8.65(d，1H，J＝2.2Hz)，4.40～4.30(m，2H)，4.10～4.00(m，1H)，3.90～3.80(m，2H)，2.89(m，3H)
Example 87
Synthesis of 2- ((8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) ethanol
(a) Synthesis of 2- ((2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) amino) ethanol
2,3, 7-trichloropyrido [2,3-b ] pyrazine (500.0mg, 2.14mmol) was dissolved in DCM (20.0mL) and ethanolamine (135.0. mu.L, 2.25mmol) and TEA (965.0. mu.L, 6.42mmol) were added thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, diluted with DCM, and then stirred at room temperature for 1 hour. The solid formed was filtered and dried under reduced pressure to give the unpurified brown solid compound 2- ((2, 7-dichloropyrido [2,3-b ] pyrazin-3-yl) amino) ethanol.
LC/MS ESI(+)：259(M+1)，261(M+3)
(b) Synthesis of 2- ((7-chloro-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) amino) ethanol
Hydrazine monohydrate (276.0. mu.L, 8.79mmol) was added to unpurified 2- ((2, 7-dichloropyrido [2,3-b]Pyrazin-3-yl) amino) ethanol and EtOH (30.0mL), then stirred at room temperature for 4 hours. With Et2The reaction mixture was diluted with O and then stirred at room temperature for 1 hour. The solid formed was filtered and then dried under reduced pressure to give the unpurified brown solid compound 2- ((7-chloro-2-hydrazinopyrido [2, 3-b)]Pyrazin-3-yl) amino) ethanol.
LC/MS ESI(+)：255(M+1)，257(M+3)
(c) Synthesis of 2- ((8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) ethanol
Unpurified 2- ((7-chloro-2-hydrazinopyrido [2, 3-b)]Pyrazin-3-yl) amino) ethanol and trimethyl orthoformate (16.0mL) were stirred at 80 ℃ for 4 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure and then dissolved in MeOH. EtOAc was added to the mixture and the mixture was diluted. The organic layer was washed with water and brine, and anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. EtOAc (20.0mL) was added to the residue and stirred for 1 hour. The resulting solid was filtered and dried under reduced pressure to give 2- ((8-chloropyrido [2, 3-e) as a white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) amino) ethanol (82.0mg, 14%, 3 steps).
LC/MS ESI(+)：265(M+1)，267(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.93(s，1H)，8.82(d，1H，J＝2.7Hz)，8.64-8.58(brs，1H)，8.55(d，1H，J＝2.7Hz)，4.84(m，1H)，3.70-3.64(m，4H)
Example 88
Synthesis of 1- (8-chloroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
1- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (180.0mg, 0.47mmol) was dissolved in 2, 2-diethoxyethylamine (2.0mL), stirred at room temperature for 12 hours, to which EtOAc solvent was added. The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering and then distilling under reduced pressure to obtain a yellow solid compound (1- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
(b) Synthesis of 1- (8-chloroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
Unpurified (1- (7-chloro-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (225.1mg, 0.47mmol) and 4-methylbenzenesulfonic acid (1160.0mg, 6.08mmol) were dissolved in IPA (3.0mL), stirred at 100 deg.C for 2 hours, and then cooled to room temperature. Water was added to the reaction mixture. With saturated NaHCO3The resulting solution was basified with aqueous solution (pH 9) and then extracted with DCM. With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-chloroimidazo [1,2-a ]]Pyrido [2,3-e ]]Pyrazin-4-yl) -N-methylazetidin-3-amine (70.0mg, 52%).
LC/MS ESI(+)：289(M+1)，291(M+3)
1H-NMR(300MHz，CDCl3)；：8.53(m，1H)，7.94-7.89(m，2H)，7.66(m，1H)，5.12-4.13(m，4H)，3.83(m，1H)，2.49(s，3H)
Example 89
Synthesis of 1- (8-bromoimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (7-bromo-2- ((2, 2-diethoxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
Reacting (1- (7-bromo-2-chloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (177.0mg, 0.41mmol) was dissolved in 2, 2-diethoxyethylamine (2.0mL) and the mixture was stirred at room temperature for 12 h. EtOAc solvent was then added to the reaction mixture, washed with brine, and dried over anhydrous Na2SO4Drying, filtering and then distilling under reduced pressure to obtain a yellow solid compound (1- (7-bromo-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester.
(b) Synthesis of 1- (8-bromoimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
Unpurified (1- (7-bromo-2- ((2, 2-diethoxyethyl) amino) pyrido [2, 3-b)]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (215.0mg, 0.41mmol) and 4-methylbenzenesulfonic acid (1020.0mg, 5.36mmol) were dissolved in IPA (3.0mL), and the mixture was stirred at 100 deg.C for 2 hours and then cooled to room temperature. Water was added to the reaction mixture and saturated NaHCO was used3The aqueous solution was basified (pH 9) and then extracted with DCM. With anhydrous Na2SO4The organic layer was dried, filtered, distilled under reduced pressure, and then purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain a milky white solid compound 1- (8-bromoimidazo [1,2-a ]]Pyrido [2,3-e ]]Pyrazin-4-yl) -N-methylazetidin-3-amine (65.0mg, 47%).
LC/MS ESI(+)：333(M+1)，335(M+3)
1H-NMR(300MHz，CDCl3)；：8.60(d，1H，J＝2.1Hz)，8.06(d，1H，J＝2.1Hz)，7.89(d，1H，J＝1.5Hz)，7.65(d，1H，J＝1.5Hz)，5.08-4.20(m，4H)，3.82(m，1H)，2.48(s，3H)
Example 90
Synthesis of tert-butyl (1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate
(a) Synthesis of methyl 2, 5-dichloronicotinate
2, 5-Dichloronicotinic acid (2150.0mg, 11.20mmol) was dissolved in a mixed solution of DCM (10.0mL) and MeOH (5.0mL), to which trimethylsilyldiazomethane (2.0M in hexane) (11.2mL, 22.40mmol) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 9). The product-containing fractions were collected and evaporated to give methyl 2, 5-dichloronicotinate as a white solid (2020.0mg, 87%).
LC/MS ESI(+)：206(M+1)
1H-NMR(300MHz，CDCl3)；：8.48(d，1H，J＝2.6Hz)，8.16(d，1H，J＝2.6Hz)，3.98(s，3H)
(b) Synthesis of methyl 5-chloro-2- ((4-methoxybenzyl) amino) nicotinate
EtOH (30.0mL) was added to methyl 2, 5-dichloronicotinate (2020.0mg, 9.79mmol) and 4-methoxybenzylamine (2.6mL, 19.60mmol) was added thereto. The mixture was stirred at 65 ℃ for 12 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 9). The product-containing fractions were collected and evaporated to give methyl 5-chloro-2- ((4-methoxybenzyl) amino) nicotinate (840.0mg, 28%) as a colourless liquid.
LC/MS ESI(+)：307(M+1)
(c) Synthesis of 6-chloro-1- (4-methoxybenzyl) -1H-pyrido [2,3-d ] [1,3] oxazine-2, 4-dione
Methyl 5-chloro-2- ((4-methoxybenzyl) amino) nicotinate (205.0mg, 0.67mmol) was dissolved in 1, 4-dioxane (3.0mL) and diphosgene (120.0 μ L, 1.01mmol) was added slowly thereto at room temperature. The reaction mixture was stirred at 110 ℃ for 12 hours. The reaction mixture was then distilled under reduced pressure. The residue was dissolved in toluene (5.0mL), distilled under reduced pressure, and then dried to give 6-chloro-1- (4-methoxybenzyl) -1H-pyrido [2,3-d ] [1,3] oxazine-2, 4-dione as a pale yellow solid.
LC/MS ESI(+)：319(M+1)
(d) Synthesis of 6-chloro-4-hydroxy-1- (4-methoxybenzyl) -3-nitro-1, 8-naphthyridin-2 (1H) -one
DMA (3.0mL) was added to NaH (40.4mg: in 60% oil, 1.01mmol), to which was then added ethyl nitroacetate (110.0. mu.L, 1.01 mmol). The mixture was stirred at room temperature for 30 minutes. Reacting 6-chloro-1- (4-methoxybenzyl) -1H-pyrido [2,3-d ] at room temperature][1,3]A solution of oxazine-2, 4-dione in DMA (1.0mL) was slowly added to the reaction mixture. The reaction mixture was stirred at 110 ℃ for 12 hours, cooled to room temperature, and then filtered through celite. The filtrate was distilled under reduced pressure. The residue was dissolved in toluene (5.0mL) and distilled under reduced pressure. In DCM and Et2The solid formed was stirred in a mixture of O and then filtered to give the compound 6-chloro-4-hydroxy-1- (4-methoxybenzyl) -3-nitro-1, 8-naphthyridin-2 (1H) -one (232.0mg, 95%, 2 steps) as a dark yellow solid.
LC/MS ESI(+)：362(M+1)
(e) Synthesis of 8-chloro-5- (4-methoxybenzyl) -2-methyl-oxazolo [4,5-c ] [1,8] naphthyridin-4 (5H) -one
A mixture of 6-chloro-4-hydroxy-1- (4-methoxybenzyl) -3-nitro-1, 8-naphthyridin-2 (1H) -one (200.0mg, 0.55mmol), Zn (220.0mg, 3.36mmol), acetic anhydride (2.7mL, 28.30mmol) and acetic acid (4.0mL) was stirred at 120 ℃ for 12H. The reaction mixture was cooled to room temperature and then filtered through celite. The filtrate was concentrated under reduced pressure, to which TEA (0.5mL, purified by silica gel column chromatography (EtOAc: n-Hex ═ 1:1) was added, fractions containing the product were collected and evaporated to give the compound 8-chloro-5- (4-methoxybenzyl) -2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4 (5H) -one (108.0mg, 52%) as a white solid.
LC/MS ESI(+)：356(M+1)
(f) Synthesis of 4, 8-dichloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridine
Reacting 8-chloro-5- (4-methoxybenzyl) -2-methyl oxazolo [4,5-c][1,8]Naphthyridin-4 (5H) -one (108.0mg, 0.30mmol) in POCl3(2.0mL), the mixture was stirred at 140 ℃ for 12 hours. The reaction mixture was cooled to 0 ℃ and poured over saturated Na with crushed ice2CO3Aqueous solution, then extracted with DCM. With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 3). Collecting the product-containing fractions, and evaporating to obtain 4, 8-dichloro-2-methyloxazolo [4,5-c ] as a pale yellow solid][1,8]Naphthyridine (42.0mg, 54%).
LC/MS ESI(+)：254(M+1)
1H-NMR(300MHz，CDCl3)；：9.04(d，1H，J＝2.7Hz)，8.49(d，1H，J＝2.7Hz)，2.85(s，3H)
(g) Synthesis of tert-butyl (1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate
DMF (2.0mL) was added to 4, 8-dichloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridine (19.4mg, 0.08mmol), to which was then added azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (26.0mg, 0.12mmol) and TEA (30.0. mu.L, 0.23 mmol). The reaction mixture was stirred for 6 hours and distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 2). The product-containing fractions were collected and evaporated to give the compound tert-butyl (1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate as a pale yellow solid (30.0mg, 97%).
LC/MS ESI(+)：404(M+1)
1H-NMR(300MHz，CDCl3)；：8.73(d，1H，J＝2.7Hz)，8.19(d，1H，J＝2.7Hz)，5.41-4.91(m，1H)，4.82-4.68(m，2H)，4.59-4.47(m，2H)，2.99(s，3H)，2.71(s，3H)，1.48(s，9H)
Example 91
Synthesis of 1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate (30.0mg, 0.07mmol) was dissolved in DCM (2.0mL), to which was added TFA (1.0mL, 13.50 mmol). The mixture was stirred for 6 hours. The reaction mixture was distilled under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 50). The product-containing fractions were collected and evaporated to give the compound 1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine (20.3mg, 87%) as a pale yellow solid.
LC/MS ESI(+)：304(M+1)
1H-NMR(300MHz，CDCl3)；：8.71(d，1H，J＝2.7Hz)，8.16(d，1H，J＝2.7Hz)，4.78-4.68(m，2H)，4.29-4.19(m，2H)，3.87-3.76(m，1H)，2.71(s，3H)，2.48(s，3H)
Example 92
Synthesis of 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) oxazolo [4,5-c ] [1,8] naphthyridine
4, 8-dichloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridine (10.0mg, 0.04mmol) was added to DMF (1.0mL), to which was added N-methylpiperazine (26.0mg, 0.12mmol) and TEA (30.0. mu.L, 0.23 mmol). The reaction mixture was stirred for 12 hours and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 10). The product-containing fractions were collected and evaporated to give the compound 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) oxazolo [4,5-c ] [1,8] naphthyridine as a pale yellow solid (7.9mg, 83%).
LC/MS ESI(+)：318(M+1)
1H-NMR(300MHz，CDCl3)；：8.72(d，1H，J＝2.7Hz)，8.17(d，1H，J＝2.7Hz)，4.51-4.28(m，4H)，2.72(s，3H)，2.62-2.53(m，4H)，2.36(s，3H)
Example 93
Synthesis of 1- (8-chloropyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-chloropyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Adding NaN3(122.0mg, 1.87mmol) was added to (1- (2, 7-dichloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (80.0mg, 0.21mmol) and EtOH (2.1mL) in suspension. The reaction mixture was stirred at 70 ℃ for 12 hours, cooled to room temperature and then concentrated under reduced pressure. Water was added thereto to form a solid, and the formed solid was dried under reduced pressure to give a white solid compound (1- (8-chloropyrido [2, 3-e)]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (75.0mg, 93%).
LC/MS ESI(+)：391(M+1)，393(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.88(d，1H，J＝2.4Hz)，8.78(d，1H，J＝2.4Hz)，5.21-4.85(m，2H)，4.82(m，1H)，4.46(m，1H)，4.45(m，1H)，2.93(s，3H)，1.42(s，9H)
(b) Synthesis of 1- (8-chloropyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (8-chloropyrido [2, 3-e)]Tetrazolo [1,5-a ] s]Mixture of pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (75.0mg, 0.19mmol) and DCM (0.6mL)Then, the mixture was stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated. Adding Et2O is added to the residue to form a solid, the formed solid is filtered and then dried under reduced pressure to obtain 1- (8-chloropyrido [2, 3-e) as a white solid]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) -N-methylazetidin-3-amine (42.0mg, 75%).
LC/MS ESI(+)：291(M+1)，293(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.83(d，1H，J＝2.4Hz)，8.76(d，1H，J＝2.4Hz)，4.90(m，1H)，4.60-4.39(m，2H)，4.05(m，1H)，3.75(m，1H)，2.46(m，1H)，2.31(s，3H)
Example 94
Synthesis of 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine
Adding NaN3(55.0mg, 0.85mmol) was added to 2, 7-dichloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (28.0mg, 0.09mmol) and EtOH (1.0 mL). The mixture was stirred at 70 ℃ for 12 hours. The reaction mixture was cooled to room temperature and distilled under reduced pressure. Then, water was added thereto to form a solid, and the formed solid was filtered and dried under reduced pressure to obtain 8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] as an orange solid]Tetrazolo [1,5-a ] s]Pyrazine (21.0mg, 75%).
LC/MS ESI(+)：305(M+1)，307(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.87(d，1H，J＝2.1Hz)，8.79(d，1H，J＝2.4Hz)，4.80-3.90(m，4H)，2.58-2.50(m，4H)，2.26(s，3H)
Example 95
Synthesis of 1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Adding NaN3(71.0mg, 1.09mmol) was added to (1- (7-bromo-2-chloropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (52.0mg, 0.12mmol) and EtOH (1.2mL) in suspension. The mixture was stirred at 70 ℃ for 12 hours. The reaction mixture was cooled to room temperature and then distilled under reduced pressure. Wherein water is added to form a solid, the solid formed is filtered and dried under reduced pressure to obtain the compound (1- (8-bromopyrido [2, 3-e) as a white solid]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (45.0mg, 85%).
LC/MS ESI(+)：435(M+1)，437(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.94(d，1H，J＝2.4Hz)，8.84(d，1H，J＝2.4Hz)，5.25-4.90(m，2H)，4.82(m，1H)，4.65-4.30(m，2H)，2.94(s，3H)，1.43(s，9H)
(b) Synthesis of 1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (8-bromopyrido [2,3-e ]]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (45.0mg, 0.10mmol) and DCM (0.6 mL). The reaction mixture was stirred at room temperature for 1 hour and purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 60). The product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered, followed by drying under reduced pressure to give 1- (8-bromopyrido [2, 3-e) as a white solid]Tetrazolo [1,5-a ] s]Pyrazin-4-yl) -N-methylazetidin-3-amine (28.0mg, 80%).
LC/MS ESI(+)：335(M+1)，337(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.92(bs，1H)，8.81(bs，1H)，4.90(m，1H)，4.55-4.38(m，2H)，4.04(m，1H)，3.76(m，1H)，2.31(s，3H)
Example 96
Synthesis of 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine
Adding NaN3(50.0mg, 0.76mmol) was added to 7-bromo-2-chloro-3- (4-methylpiperazin-1-yl) pyrido [2,3-b]Pyrazine (29.0mg, 0.08mmol) and EtOH (1.0 mL). The reaction mixture was stirred at 70 ℃ for 12 hours, then cooled to room temperature and distilled under reduced pressure. Adding water thereto to form a solid, filtering the solid formed, and drying under reduced pressure to obtain 8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e as an orange solid]Tetrazolo [1,5-a ] s]Pyrazine (24.0mg, 80%).
LC/MS ESI(+)：349(M+1)，351(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.95(d，1H，J＝2.4Hz)，8.85(d，1H，J＝2.4Hz)，4.65-4.05(m，4H)，2.60-2.50(m，4H)，2.26(s，3H)
Example 97
Synthesis of 1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of 5-chloro-3-fluoro-2-nitropyridine
5-chloro-3-fluoropyridin-2-amine (500.0mg, 3.41mmol) was dissolved in H2SO4(1.5mL), to which Na was added2S2O8(406.1mg, 1.71 mmol). The mixture was stirred at room temperature for 12 hours, and then water was poured into the reaction mixture. With saturated NaHCO3The resulting solution was basified with aqueous solution (pH 9) and then extracted with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 10: 90). The product-containing fractions were collected and evaporated to give 5-chloro-3-fluoro-2-nitropyridine (200.0mg, 33%) as a white solid.
1H-NMR(300MHz，DMSO-d6)；：8.21(d，1H，J＝1.8Hz)，8.17(m，1H)
(b) Synthesis of 5-chloro-3-hydrazino-2-nitropyridine
5-chloro-3-fluoro-2-nitropyridine (190.0mg, 1.08mmol) and hydrazine monohydrate (80.0. mu.L, 1.61mmol) were dissolved in EtOH (3.0mL) and the mixture was stirred at room temperature for 2 hours. Et was added thereto2O to form a solid, and the solid formed was filtered to give the compound 5-chloro-3-hydrazino-2-nitropyridine (170.0mg, 84%) as an orange solid.
1H-NMR(300MHz，DMSO-d6)；：9.06(brs，1H)，8.10(m，1H)，7.76(d，1H，J＝2.4Hz)
(c) Synthesis of (Z) -N' - (5-chloro-2-nitropyridin-3-yl) acetyl hydrazone amide
5-chloro-3-hydrazino-2-nitropyridine (100.0mg, 0.53mmol) and ethyl acetimidate hydrochloride (102.2mg, 0.80mmol) were added to pyridine (1.8mL) and stirred at room temperature for 3 hours. Saturated Na2CO3Aqueous solution was added to the reaction mixture, which was then extracted with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex 50: 50). The product-containing fractions were collected and evaporated to yield the compound (Z) -N' - (5-chloro-2-nitropyridin-3-yl) acetyl hydrazone amide (77.0mg, 63%) as a red solid.
1H-NMR(300MHz，DMSO-d6)；：9.53(brs，1H)，7.88(d，1H，J＝2.4Hz)，7.78(d，1H，J＝2.4Hz)，6.62(brs，2H)，1.92(s，3H)
(d) Synthesis of ethyl 1- (5-chloro-2-nitropyridin-3-yl) -3-methyl-1H-1, 2, 4-triazole-5-carboxylate
(Z) -N' - (5-chloro-2-nitropyridin-3-yl) acetyl hydrazone amide (60.0mg, 0.26mmol) and ethyl 2-chloro-2-oxoacetate (54.0. mu.L, 0.52mmol) were dissolved in Et2O (0.6mL), stirred at room temperature for 1 hour, and then toluene (6.0mL) was added thereto. The mixture was stirred at 80 ℃ for 1 hour, at 180 ℃ for 2 hours, then cooled to room temperature, water and EtOAc were added, then basified with aqueous KOH (pH 12) and extracted with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered and distilled under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc: n-Hex 50: 50). The product-containing fractions were collected and evaporated to yield the compound ethyl 1- (5-chloro-2-nitropyridin-3-yl) -3-methyl-1H-1, 2, 4-triazole-5-carboxylate (78.0mg, 56%) as a red solid.
LC/MS ESI(+)：312(M+1)，314(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.95(d，1H，J＝2.4Hz)，8.83(d，1H，J＝2.4Hz)，4.28-4.21(m，2H)，2.40(s，3H)，1.20-1.15(m，3H)
(e) Synthesis of 8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4 (5H) -one
Ethyl 1- (5-chloro-2-nitropyridin-3-yl) -3-methyl-1H-1, 2, 4-triazole-5-carboxylate (62.0mg, 0.20mmol) and Fe (166.6mg, 2.98mmol) were added to acetic acid (13.0mL), stirred at 90 ℃ for 2 hours, then cooled to room temperature. Aqueous 1N HCl was added to the reaction mixture to dissolve completely, then extracted with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered, and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). Collecting the product-containing fractions, and evaporating to obtain 8-chloro-2-methylpyrido [2,3-e ] as a white solid][1,2,4]Triazolo [1,5-a]Pyrazin-4 (5H) -one (40.0mg, 86%).
LC/MS ESI(+)：236(M+1)，238(M+3)
1H-NMR(300MHz，DMSO-d6)；：12.89(brs，1H)，8.52(d，1H，J＝2.4Hz)，8.44(d，1H，J＝2.4Hz)，2.53(s，3H)
(f) Synthesis of 4, 8-dichloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine
Reacting 8-chloro-2-methylpyrido [2,3-e ]][1,2,4]Triazolo [1,5-a]Pyrazin-4 (5H) -one (40.0mg, 0.17mmol) in POCl3(1.0mL), to which DIPEA (60.0. mu.L, 0.34mmol) was added. The reaction mixture was refluxed for 5 hours and then cooled to room temperature. The reaction mixture was poured into ice water and saturated NaHCO3Aqueous solution was neutralized and then extracted with EtOAc. The organic layer was washed with brine, anhydrous Na2SO4Drying, filtering, and distilling under reduced pressure to obtain milky white solid compound 4, 8-dichloro-2-methylpyrido [2,3-e][1,2,4]Triazolo [1,5-a]Pyrazine (43.0mg, 100%).
LC/MS ESI(+)：254(M+1)，256(M+3)
1H-NMR(300MHz，DMSO-d6)；：9.03(m，1H)，8.94(m，1H)，2.67(s，3H)
(g) Synthesis of 1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
4, 8-dichloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine (10.0mg, 0.04mmol) and TFA (16.5. mu.L, 0.12mmol) were dissolved in DMF (0.2mL), to which was added azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (13.0mg, 0.06 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate as a white solid. Unpurified tert-butyl (1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate was dissolved in DCM (1.0mL), to which was added TFA (0.4 mL). The reaction mixture was stirred at room temperature for 1 hour, then purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound 1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (7.0mg, 45%) as a white solid.
LC/MS ESI(+)：304(M+1)，306(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.60(d，1H，J＝2.4Hz)，8.45(d，1H，J＝2.4Hz)，4.83(m，1H)，4.50-4.30(m，2H)，4.00(m，1H)，3.69(m，1H)，2.57(s，3H)，2.30(brs，3H)
Example 98
Synthesis of 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine
4, 8-dichloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine (10.0mg, 0.04mmol) and TFA (16.5. mu.L, 0.12mmol) were dissolved in DMF (0.2mL), to which was added N-methylpiperazine (6.5. mu.L, 0.06 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated and purified by silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and evaporated to yield the compound 8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine as a white solid (8.0mg, 66%).
LC/MS ESI(+)：318(M+1)，320(M+3)
1H-NMR(300MHz，DMSO-d6)；：8.65(d，1H，J＝2.4Hz)，8.49(d，1H，J＝2.4Hz)，4.45-4.18(m，4H)，2.59(s，3H)，2.52(m，4H)，2.25(s，3H)
Example 99
Synthesis of 1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of 7-bromo-6-methylpyrido [2,3-b ] pyrazine-2, 3-diol
A mixture of 5-bromo-6-methylpyridine-2, 3-diamine (500.0mg, 2.47mmol) and diethyl oxalate (3.0mL) was stirred at 100 ℃ for 12 hours and then cooled to room temperature. Adding Et2O was added to the reaction mixture to form a solid. Then the solid formed was filtered and dried under reduced pressure to give 7-bromo-6-methylpyrido [2,3-b as a light brown solid]Pyrazine-2, 3-diol (611.0mg, 97%).
LC/MS ESI(+)：256(M+1)，258(M+3)
1H-NMR(400MHz，DMSO-d6)；：12.40(s，1H)，11.94(s，1H)，7.53(s，1H)，2.48(s，3H)
(b) Synthesis of 7-bromo-2, 3-dichloro-6-methylpyrido [2,3-b ] pyrazine
Reacting 7-bromo-6-methylpyrido [2,3-b ]]Pyrazine-2, 3-diol (300.0mg, 1.17mmol) and POCl3(6.0mL) of the mixture was stirred at 95 ℃ for 12 hours, then cooled to room temperature. The reaction mixture was poured into ice water to form a solid. Filtering the solid formed, drying under reduced pressure to obtain black solid compound 7-bromo-2, 3-dichloro-6-methylpyrido [2,3-b]Pyrazine (340.0mg, 99%).
LC/MS ESI(+)：292(M+1)，294(M+3)，296(M+5)
1H-NMR(400MHz，CDCl3)；：8.53(s，1H)，2.95(s，3H)
(c) Synthesis of tert-butyl (1- (7-bromo-2-chloro-6-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
TEA (140.0. mu.L, 1.02mmol) was added to a mixture of 7-bromo-2, 3-dichloro-6-methylpyrido [2,3-b ] pyrazine (100.0mg, 0.34mmol), azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (76.0mg, 0.34mmol), and DCM (3.4mL) at 0 ℃ and stirred for 1 hour. The reaction mixture was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (7-bromo-2-chloro-6-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate as a red solid (43.0mg, 28%).
LC/MS ESI(+)：442(M+1)，444(M+3)
1H-NMR(400MHz，CDCl3)；：8.23(s，1H)，5.02-4.90(m，1H)，4.78-4.68(m，2H)，4.50-4.49(m，2H)，2.96(s，3H)，2.836(s，3H)，1.48(s，9H)
(d) Synthesis of tert-butyl (1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Hydrazine monohydrate (15.0 μ L, 0.49mmol) was added to a suspension of tert-butyl (1- (7-bromo-2-chloro-6-methylpyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (43.0mg, 0.10mmol) and EtOH (1.0 mL). The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was dissolved in trimethyl orthoformate (1.0mL) and stirred at 85 ℃ for 1 hour. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (MeOH: DCM ═ 1: 40). Fractions containing the product were collected and evaporated to give the compound tert-butyl (1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (35.0mg, 80%) as a light brown solid.
LC/MS ESI(+)：448(M+1)，450(M+3)
(e) Synthesis of 1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (8-bromo-7-methylpyrido [2, 3-e)][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (35.0mg, 0.08mmol) and DCM (0.6mL) was stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered, followed by drying under reduced pressure to give 1- (8-bromo-7-methylpyrido [2, 3-e) as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (12.0mg, 43%).
LC/S ESI(+)：348(M+1)，350(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.93(s，1H)，8.63(s，1H)，4.90(m，1H)，4.45-4.35(m，2H)，3.97(m，1H)，3.71(m，1H)，2.60(s，3H)，2.30(s，3H)
Example 100
Synthesis of 8-bromo-7-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
(a) Synthesis of 7-bromo-2-chloro-6-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine
TEA (140.0. mu.L, 1.02mmol) was added to a mixture of 7-bromo-2, 3-dichloro-6-methylpyrido [2,3-b ] pyrazine (100.0mg, 0.34mmol), N-methylpiperazine (38.0. mu.L, 0.34mmol) and DCM (3.4mL) obtained from example 99 step (b) at 0 ℃ and then stirred for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and evaporated to yield the compound 7-bromo-2-chloro-6-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine as a red solid (23.0mg, 19%).
LC/MS ESI(+)：356(M+1)，358(M+3)，360(M+5)
1H-NMR(400MHz，DMSO-d6)；：8.32(s，1H)，3.79-3.70(m，4H)，2.87(s，3H)，2.65-2.62(m，4H)，2.38(s，3H)
(b) Synthesis of 8-bromo-7-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Hydrazine monohydrate (10.0 μ L, 0.32mmol) was added to a suspension of 7-bromo-2-chloro-6-methyl-3- (4-methylpiperazin-1-yl) pyrido [2,3-b ] pyrazine (23.0mg, 0.06mmol) and EtOH (1.0 mL). The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was dissolved in trimethyl orthoformate (1.0mL) and stirred at 85 ℃ for 1 hour. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to yield the compound 8-bromo-7-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine as a yellow solid (1.1mg, 4%).
LC/MS ESI(+)：362(M+1)，364(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.99(s，1H)，8.93(s，1H)，4.46-4.30(m，4H)，3.32(s，3H)，2.63(s，3H)2.54-2.50(m，4H)
Example 101
Synthesis of 8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-7-ol HCl salt
(a) Synthesis of tert-butyl (1- (8-chloro-7-hydroxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
CsF (32.2mg, 0.21mmol) was added to a mixture of tert-butyl (1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (30.0mg, 0.07mmol) obtained from example 27 step (b) and NMP (1.0 mL). The reaction mixture was stirred at 135 ℃ for 12 hours and then cooled to room temperature. CsF (161.0mg, 1.06mmol) was added thereto and stirred at 145 ℃ for 12 hours. The reaction mixture was cooled to room temperature and purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give tert-butyl (1- (8-chloro-7-hydroxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate as a cream solid (10.0mg, 35%).
LC/MS ESI(+)：406(M+1)，408(M+3)
(b) Synthesis of 8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-7-ol HCl salt
Reacting (1- (8-chloro-7-hydroxypyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]A suspension of t-butyl pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (9.0mg, 0.02mmol) and 4N HCl in 1, 4-dioxane (0.5mL) was stirred at room temperature for 1 hour. Adding Et2O was added to the reaction mixture to form a solid. The solid formed was filtered and dried under reduced pressure to give 8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-7-ol HCl salt (3.2mg, 43%).
LC/MS ESI(+)：306(M+1)，308(M+3)
1H-NMR(400MHz，DMSO-d6)；：13.00-12.30(m，1H)，9.80(s，1H)，9.75-9.65(m，2H)，8.76(s，1H)，5.02(m，1H)，4.89(m，1H)，4.53(m，2H)，4.23(m，1H)，2.63(s，3H)
Example 102
Synthesis of N- (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine
(a) Synthesis of O-benzoyl-N- (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine
Mixing CHCl3(5.0mL) was added to 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (120.0mg, 0.36mmol), benzoyl peroxide (116.0mg, 0.36mmol) and K2CO3(174.0mg, 1.26mmol) was stirred at 65 ℃ for 12 hours. The reaction mixture was cooled to room temperature, filtered through celite,then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 98:2 to 93: 7). The product-containing fractions were collected and evaporated to give the gray solid compound O-benzoyl-N- (1- (8-bromopyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine (150.0mg, 92%).
LC/MS ESI(+)：454(M+1)，456(M+3)
(b) Synthesis of N- (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine
NaOH (24.0mg, 0.60mmol) was dissolved in MeOH (8.0mL), to which O-benzoyl-N- (1- (8-bromopyrido [2,3-e ] was added][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine (120.0mg, 0.26 mmol). The mixture was stirred for 12 hours and purified by silica gel column chromatography (DCM: MeOH ═ 91: 9). The product-containing fractions were collected and evaporated. The residue was dissolved in DMSO and then purified by reverse phase high performance chromatography (ACN: H)2O65: 35). The product containing fractions were collected and evaporated to give a light brown solid. The solid obtained is taken up in Et2Stirring in O for 30 min, and filtering to obtain light brown solid compound N- (1- (8-bromopyrido [2, 3-e))][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine (71.0mg, 77%).
LC/MS ESI(+)：350(M+1)，352(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.95(s，1H)，8.90(d，1H，J＝2.2Hz)，8.59(d，1H，J＝2.2Hz)，4.76(m，2H)，4.30(m，2H)，3.77(m，1H)，2.51(s，3H)
Example 103
Synthesis of 1- (2-bromopyrido [3,2-e ] pyrrolo [1,2-c ] pyrimidin-6-yl) -N-methylazetidin-3-amine
(a) Synthesis of 2-bromopyrido [3,2-e ] pyrrolo [1,2-c ] pyrimidin-6-ol
Pd (PPh)3)4(193.0mg, 0.17mmol) was added to 5-bromo-3-iodopyridin-2-amine (500.0mg, 1.67mmol), (1- (tert-butoxycarbonyl) -1H-pyrrol-2-yl) boronic acid (423.0mg, 2.00mmol), Cs2CO3(1600.0mg，5.01mmol)、H2O (4.0mL) and DME (16.0 mL). The mixture was reacted in a microwave reactor at 100W at 110 ℃ for 2 hours and then cooled to room temperature. Adding Et2O was added to the reaction mixture to form a solid. The solid formed was filtered and then dried under reduced pressure to form 2-bromopyrido [3,2-e ] as a white solid]Pyrrolo [1,2-c]Pyrimidin-6-ol (180.0mg, 41%).
LC/MS ESI(+)：264(M+1)，266(M+3)
1H-NMR(400MHz，DMSO-d6)；：12.11(s，1H)，8.66(d，1H，J＝1.5Hz)，8.39(d，1H，J＝1.5Hz)，7.66(m，1H)，7.23(m，1H)，6.72(m，1H)
(b) Synthesis of tert-butyl (1- (2-bromopyrido [3,2-e ] pyrrolo [1,2-c ] pyrimidin-6-yl) azetidin-3-yl) (methyl) carbamate
DIPEA (24.0. mu.L, 0.14mmol) was added to 2-bromopyrido [3,2-e ]]Pyrrolo [1,2-c]Pyrimidin-6-ol (30.0mg, 0.11mmol) and POCl3(1.1mL) and then stirred at 120 ℃ for 20 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was dissolved in DCM (1.1mL) and tert-butyl azetidin-3-yl (methyl) carbamate HCl salt (50.0mg, 0.23mmol) and DIPEA (200.0. mu.L, 1.13mmol) were added thereto. The mixture was stirred at room temperature for 12 hours. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 50). The product-containing fractions were collected and evaporated to give the compound (1- (2-bromopyrido [3, 2-e) as a yellow solid]Pyrrolo [1,2-c]Pyrimidin-6-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (28.0mg, 57%).
LC/MS ESI(+)：433(M+1)，435(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.65(d，1H，J＝1.8Hz)，8.46(d，1H，J＝1.8Hz)，7.60(m，1H)，7.25(m，1H)，6.84(m，1H)，5.01-4.90(m，1H)，4.68(m，2H)，4.54(m，2H)，2.93(s，3H)，1.41(s，9H)
(c) Synthesis of 1- (2-bromopyrido [3,2-e ] pyrrolo [1,2-c ] pyrimidin-6-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (2-bromopyrido [3,2-e ]]Pyrrolo [1,2-c]To a mixture of t-butyl pyrimidin-6-yl) azetidin-3-yl) (methyl) carbamate (28.0mg, 0.06mmol) and DCM (0.6mL) was stirred at room temperature for 1 hour. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1:40), and the product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered, followed by drying under reduced pressure to give 1- (2-bromopyrido [3, 2-e) as a yellow solid]Pyrrolo [1,2-c]Pyrimidin-6-yl) -N-methylazetidin-3-amine (14.0mg, 64%).
LC/MS ESI(+)：332(M+1)，334(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.62(d，1H，J＝1.8Hz)，8.44(d，1H，J＝1.8Hz)，7.60(m，1H)，7.25(m，1H)，6.82(m，1H)，4.61(m，2H)，4.17(m，2H)，3.60(m，1H)，2.38(m，1H)，2.37(s，3H)
Example 104
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2,2, 2-trifluoroacetate
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2,2, 2-trifluoroacetate
Tert-butyl (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (300.0mg, 0.69mmol) was dissolved in DCM (1.8mL), to which TFA (0.9mL) was slowly added at room temperature. The reaction mixture was stirred at room temperature for 30 minutes, then concentrated under reduced pressure. MeOH (1.84mL) was added to form a solid. The solid formed was filtered and dried to give the compound 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2,2, 2-trifluoroacetate as a white solid (270.0mg, 90%).
LC/MS ESI(+)：334(M+1)
1H-NMR(400MHz，DMSO-d6)；：10.03(s，1H)，9.31(s，1H)，8.99(d，1H，J＝1.5Hz)，8.67(d，1H，J＝1.5Hz)，5.04(m，1H)，4.85(m，1H)，4.59(m，1H)，4.42(m，1H)，4.27(m，1H)，2.69(s，3H)
Example 105
Synthesis of (S) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (S) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (500.0mg, 1.17mmol) and (S) -2-aminopropan-1-ol (193.0mg, 2.57mmol) in EtOH (20.0mL) was stirred at 80 ℃ for 8 hours. The reaction mixture was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH 98:2-95: 5). The product-containing fractions were collected and evaporated to yield the compound (S) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester as a yellow solid (349.0mg, 64%).
LC/MS ESI(+)：467(M+1)，469(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.40(d，1H，J＝2.4Hz)，7.97(d，1H，J＝2.4Hz)，6.18(d，1H，J＝7.8Hz)，4.85(brs，1H)，4.80(t，1H，J＝5.6Hz)，4.48(q，2H，J＝8.4Hz)，4.20-4.35(m，3H)，3.50(m，2H)，2.88(s，3H)，1.40(s，9H)，1.19(d，3H，J＝6.6Hz)
(b) Synthesis of (S) - (tert-butyl 1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (S) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (199.0mg, 0.43mmol), MsCl (63.0mg, 0.56mmol), and TEA (129.0mg, 1.29mmol) in DCM (10.0mL) was stirred at room temperature for 2 hours, then concentrated under reduced pressure. The residue was purified by column chromatography over silica gel (DCM: MeOH ═ 98:2-95: 5). The product-containing fractions were collected and evaporated to give the compound (S) - (tert-butyl 1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (211.0mg, quantitative) as a yellow solid.
LC/MS ESI(+)：449(M+1)，451(M+3)
(c) Synthesis of (S) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (S) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (100.0mg, 0.22mmol) was dissolved in DCM (1.0mL) and TFA (1.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure. The residue was neutralized with TEA and purified by amine silica gel and silica gel column chromatography (DCM: MeOH ═ 90: 10). The product-containing fractions were collected and evaporated to yield the compound (S) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (42.0mg, 54%) as a yellow solid.
LC/MS ESI(+)：349(M+1)，351(M+3)
1H-NMR(400MHz，DMSO-d6)；：7.95(d，1H，J＝2.2Hz)，7.27(d，1H，J＝2.2Hz)，4.68(m，1H)，4.35(m，1H)，4.23(m，1H)，4.04(t，2H，J＝10.6Hz)，3.80(m，1H)，3.45-3.57(m，2H)，2.23(s，3H)，1.25(d，3H，J＝6.7Hz)
Example 106
Synthesis of (R) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (R) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (500.0mg, 1.17mmol) and (R) -2-aminopropan-1-ol (193.0mg, 2.57mmol) in EtOH (20.0mL) was stirred at 80 ℃ for 8 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH 98:2-95: 5). The product-containing fractions were collected and evaporated to yield the compound (R) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester as a yellow solid (378.0mg, 69%).
LC/MS ESI(+)：467(M+1)，469(M+3)
(b) Synthesis of (R) - (tert-butyl 1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (R) - (1- (7-bromo-2- ((1-hydroxypropan-2-yl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (378.0mg, 0.81mmol), MsCl (120.0mg, 1.05mmol), and TEA (246.0mg, 2.43mmol) in DCM (10.0mL) was stirred at room temperature for 12 hours, then concentrated under reduced pressure. The residue was purified by column chromatography over silica gel (DCM: MeOH ═ 98:2-95: 5). The product-containing fractions were collected and evaporated to yield the compound (R) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester as a yellow solid (320.0mg, 88%).
LC/MS ESI(+)：449(M+1)，451(M+3)
(c) Synthesis of (R) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (R) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (150.0mg, 0.33mmol) was dissolved in DCM (1.5mL) and TFA (1.5mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 3 hours, then concentrated under reduced pressure. The residue was neutralized with TEA and purified by amine silica gel and silica gel column chromatography (DCM: MeOH ═ 90: 10). The product-containing fractions were collected and evaporated to yield the compound (R) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (75.0mg, 64%) as a yellow solid.
LC/MS ESI(+)：349(M+1)，351(M+3)
1H-NMR(400MHz，DMSO-d6)；：7.95(d，1H，J＝2.2Hz)，7.27(d，1H，J＝2.2Hz)，4.68(m，1H)，4.35(m，1H)，4.22(m，1H)，4.03(t，2H，J＝10.6Hz)，3.80(m，1H)，3.44-3.58(m，2H)，2.24(s，3H)，1.25(d，3H，J＝6.7Hz)
Example 107
Synthesis of 1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (7-bromo-2- ((2-hydroxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (1- (7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (500.0mg, 1.17mmol) and 2-aminoethanol (157.0mg, 2.57mmol) in EtOH (20.0mL) was stirred at 90 ℃ for 12 hours. The reaction mixture was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH 98:2-95: 5). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (7-bromo-2- ((2-hydroxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (481.0mg, 91%) as a yellow solid.
LC/MS ESI(+)：453(M+1)，455(M+3)
(b) Synthesis of tert-butyl (1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (1- (7-bromo-2- ((2-hydroxyethyl) amino) pyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate (481.0mg, 1.06mmol), MsCl (158.0mg, 1.38mmol), and TEA (322.0mg, 3.18mmol) in DCM (10.0mL) was stirred at room temperature for 6 hours, then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (435.0mg, 94%) as a yellow solid.
LC/MS ESI(+)：435(M+1)，437(M+3)
1H-NMR(400MHz，DMSO-d6)；：7.96(d，1H，J＝2.2Hz)，7.31(d，1H，J＝2.2Hz)，4.83(brs，1H)，4.74(m，1H)，4.57(m，1H)，4.31(m，1H)，4.20(m，1H)，4.04(m，2H)，3.92(m，2H)，2.87(s，3H)，1.41(s，9H)
(c) Synthesis of 1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
Tert-butyl (1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (435.0mg, 1.00mmol) was dissolved in DCM (1.0mL), to which TFA (2.0mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 3 hours, then concentrated under reduced pressure. The residue was neutralized with TEA and then purified by column chromatography on amine silica gel (DCM: MeOH ═ 90: 10). The product-containing fractions were collected and evaporated to yield 1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine as a yellow solid (186.0mg, 56%).
LC/MS ESI(+)：335(M+1)，337(M+3)
1H-NMR(400MHz，DMSO-d6)；：7.95(d，1H，J＝2.2Hz)，7.28(d，1H，J＝2.2Hz)，4.67(m，1H)，4.23(m，2H)，4.03(m，2H)，3.91(m，2H)，3.81(m，1H)，3.53(m，1H)，2.32(brs，1H)，2.23(s，3H)
Example 108
Synthesis of 1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of tert-butyl (1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
A mixture of tert-butyl (S) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (139.0mg, 0.31mmol) and DDQ (140.0mg, 0.62mmol) in xylene (5.0mL) was stirred at 90 ℃ for 8 hours, and the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 98: 2). The product-containing fractions were collected and evaporated to give a yellow solid compound as a mixture of tert-butyl (S) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate and tert-butyl (1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate.
LC/MS ESI(+)：447(M+1)，449(M+3)
(b) Synthesis of 1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine
A mixture of (S) - (1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester and (1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester was dissolved in DCM (1.0mL) and TFA (2.0mL) was added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, then concentrated under reduced pressure. The residue was neutralized with TEA (pH 7) and purified by silica gel column chromatography (DCM: MeOH 98:2-90: 10). The product-containing fractions were collected and evaporated to yield 1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine as a yellow solid (14.0mg, 13%, 2 steps).
LC/MS ESI(+)：347(M+1)，349(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.75(d，1H，J＝2.1Hz)，8.53(d，1H，J＝2.1Hz)，8.43(s，1H)，4.85(m，1H)，4.38(m，2H)，3.96(m，1H)，3.63(m，1H)，2.38(s，3H)，2.30(s，3H)
Example 109
Synthesis of 1- (9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine
(a) Synthesis of 5-bromo-3- (3-methyl-1H-pyrazol-5-yl) pyridin-2-amine
Pd (PPh)3)4(190.0mg, 0.17mmol) was added to 5-bromo-3-iodopyridin-2-amine (500.0mg, 1.67mmol), (1- (tert-butoxycarbonyl) -3-methyl-1H-pyrazol-5-yl) boronic acid (755.0mg, 3.34mmol), Cs2CO3(1600.0mg，5.01mmol)、H2O (3.0mL) and DME (12.0 mL).The reaction mixture was reacted in a microwave reactor at 100W and 130 ℃ for 2 hours and 30 minutes, and then at 100W and 150 ℃ for 2 hours, followed by cooling to room temperature. Water was added to the reaction mixture and extracted 2 times with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated to give a brown solid compound, 5-bromo-3- (3-methyl-1H-pyrazol-5-yl) pyridin-2-amine and 9-bromo-2-methylpyrazolo [1,5-c]Pyrido [3,2-e]A mixture of pyrimidin-5-ols (78.0 mg).
(b) Synthesis of 9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-ol
Diphosgene (0.05mL) was added to 5-bromo-3- (3-methyl-1H-pyrazol-5-yl) pyridin-2-amine and 9-bromo-2-methylpyrazolo [1,5-c]Pyrido [3,2-e]Suspension (78.0mg) of a mixture of pyrimidin-5-ol and 1, 4-dioxane (1.0 mL). The reaction mixture was stirred at 110 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered and dried under reduced pressure to give the milky white compound 9-bromo-2-methylpyrazolo [1,5-c]Pyrido [3,2-e]Pyrimidin-5-ol (57.0mg, 12%, 2 steps).
LC/MS ESI(+)：279(M+1)，281(M+3)
1H-NMR(400MHz，DMSO-d6)；：12.34(s，1H)，8.76(m，1H)，8.57(m，1H)，7.17(s，1H)，2.38(s，3H)
(c) Synthesis of tert-butyl (1- (9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate
DIPEA (22.0. mu.L, 0.13mmol) was added to 9-bromo-2-methylpyrazolo [1,5-c]Pyrido [3,2-e]Pyrimidin-5-ol (30.0mg, 0.11mmol) and POCl3(2.0mL) was stirred at 120 ℃ for 20 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was dissolved in DCM (1.0 mL). To which azetidin-3-yl (methyl) amine is addedTert-butyl benzoate HCl salt (119.0mg, 0.54mmol) and DIPEA (400.0. mu.L, 2.14 mmol). The mixture was then stirred at room temperature for 12 hours. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 50). The product-containing fractions were collected and evaporated to give the compound (1- (9-bromo-2-methylpyrazolo [1, 5-c) as a yellow solid]Pyrido [3,2-e]Pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (32.0mg, 67%).
LC/MS ESI(+)：448(M+1)，450(M+3)
1H-NMR(400MHz，CDCl3)；：8.65(m，1H)，8.17(m，1H)，6.67(s，1H)，5.20-5.10(m，1H)，4.86(m，2H)，4.65(m，2H)，2.99(s，3H)，2.46(m，3H)，1.48(s，9H)
(d) Synthesis of 1- (9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (9-bromo-2-methylpyrazolo [1, 5-c)]Pyrido [3,2-e]To a mixture of t-butyl pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate (32.0mg, 0.07mmol) and DCM (0.6mL) was stirred at room temperature for 30 min. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. The solid formed was filtered and dried under reduced pressure to give the milky white compound 1- (9-bromo-2-methylpyrazolo [1, 5-c)]Pyrido [3,2-e]Pyrimidin-5-yl) -N-methylazetidin-3-amine (15.0mg, 60%).
LC/MS ESI(+)：347(M+1)，349(M+3)
1H-NMR(400MHz，CDCl3)；：8.64(d，1H，J＝1.8Hz)，8.15(d，1H，J＝1.8Hz)，6.65(s，1H)，4.82(m，2H)，4.37(m，2H)，3.75(m，1H)，2.48(s，3H)，2.47(s，3H)
Example 110
Synthesis of 1- (9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine
(a) Synthesis of 5-bromo-3- (1H-pyrazol-5-yl) pyridin-2-amine
Pd (PPh)3)4(39.0mg, 0.03mmol) was added to 5-bromo-3-iodopyridin-2-amine (100.0mg, 0.34mmol), (1- (tert-butoxycarbonyl) -1H-pyrazol-5-yl) boronic acid (99.0mg, 0.47mmol), Cs2CO3(327.0mg，1.01mmol)、H2O (0.5mL) and DME (2.0 mL). The reaction mixture was reacted in a microwave reactor at 100W and 120 ℃ for 1 hour and 30 minutes, then cooled to room temperature, and Pd (PPh) was added thereto3)4(20.0mg, 0.02mmol) and (1- (tert-butoxycarbonyl) -1H-pyrazol-5-yl) boronic acid (50.0mg, 0.24 mmol). The reaction mixture was reacted in a microwave reactor at 100W and 130 ℃ for 2 hours, and then cooled to room temperature. Water was added to the reaction mixture and extracted 2 times with DCM. With anhydrous Na2SO4The organic layer was dried, filtered, and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 50). The product-containing fractions were collected and evaporated to give a brown solid compound, 5-bromo-3- (1H-pyrazol-5-yl) pyridin-2-amine and 9-bromopyrazolo [1,5-c]Pyrido [3,2-e]A mixture of pyrimidin-5-ols (39.0 mg).
(b) Synthesis of 9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-ol
Diphosgene (0.05mL) was added to 5-bromo-3- (1H-pyrazol-5-yl) pyridin-2-amine and 9-bromopyrazolo [1,5-c ]]Pyrido [3,2-e]Suspension of a mixture of pyrimidin-5-ol and 1, 4-dioxane (1.0 mL). The reaction mixture was stirred at 110 ℃ for 1 hour and then cooled to room temperature. Et was added thereto2O to form a solid. The resulting solid was filtered and dried under reduced pressure to give 9-bromopyrazolo [1,5-c as a yellow solid]Pyrido [3,2-e]Pyrimidin-5-ol (32.0mg, 36%, 2 steps).
LC/MS ESI(+)：265(M+1)，267(M+3)
1H-NMR(400MHz，CDCl3+DMSO-d6)；：8.51(d，1H，J＝1.5Hz)，8.43(d，1H，J＝1.5Hz)，8.04(m，1H)，7.11(m，1H)
(c) Synthesis of tert-butyl (1- (9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate
DIPEA (25.0. mu.L, 0.15mmol) was added to 9-bromopyrazolo [1,5-c ]]Pyrido [3,2-e]Pyrimidin-5-ol (32.0mg, 0.12mmol) and POCl3(2.0mL) was stirred at 120 ℃ for 24 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was dissolved in DCM (1.2mL) and tert-butyl azetidin-3-yl (methyl) carbamate HCl salt (135.0mg, 0.61mmol) and DIPEA (420.0. mu.L, 2.42mmol) were added thereto. The reaction mixture was stirred at room temperature for 12 hours, then purified by amine silica gel column chromatography (EtOAc: n-Hex ═ 1: 5). The product-containing fractions were collected and evaporated to give the compound (1- (9-bromopyrazolo [1, 5-c) as a yellow solid]Pyrido [3,2-e]Pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (32.0mg, 62%).
LC/MS ESI(+)：433(M+1)，435(M+3)
1H-NMR(400MHz，CDCl3)；：8.68(m，1H)，8.24(m，1H)，7.97(m，1H)，6.89(m，1H)，5.20-5.10(m，1H)，4.89(m，2H)，4.68(m，2H)，2.99(s，3H)，1.48(s，9H)
(d) Synthesis of 1- (9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine
TFA (0.4mL) was added to (1- (9-bromopyrazolo [1, 5-c)]Pyrido [3,2-e]To a mixture of t-butyl pyrimidin-5-yl) azetidin-3-yl) (methyl) carbamate (32.0mg, 0.07mmol) and DCM (0.6mL) was stirred at room temperature for 30 min. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and evaporated. Adding Et2O was added to the residue to form a solid. Filtering the solid formed, drying under reduced pressure to obtain a milky white solid compound 1- (9-bromopyrazolo [1, 5-c)]Pyridine compoundAnd [3,2-e ]]Pyrimidin-5-yl) -N-methylazetidin-3-amine (16.0mg, 64%).
LC/MS ESI(+)：333(M+1)，335(M+3)
1H-NMR(400MHz，CDCl3)；：8.67(d，1H，J＝1.8Hz)，8.22(d，1H，J＝1.8Hz)，7.97(d，1H，J＝1.5Hz)，6.88(d，1H，J＝1.5Hz)，4.86(m，2H)，4.40(m，2H)，3.76(m，1H)，2.48(s，3H)
Example 111
Synthesis of N-methyl-1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
(a) Synthesis of 5-nitropyridine-2, 3-diamine
3, 5-dinitropyridin-2-amine (300.0mg, 1.63mmol) and ammonium sulfide solution (2.4mL, 8.15mmol) were dissolved in MeOH (11.3mL), stirred at 75 deg.C for 30 minutes, and then cooled to room temperature. The solid formed was filtered and then dried under reduced pressure to give 5-nitropyridine-2, 3-diamine (250.0mg, 99%) as a red solid.
LC/MS ESI(+)：155(M+1)
1H-NMR(400MHz，DMSO-d6)；：8.29(d，1H，J＝1.8Hz)，7.36(d，1H，J＝1.8Hz)，6.99(s，2H)，5.32(s，2H)
(b) Synthesis of 7-nitropyrido [2,3-b ] pyrazine-2, 3-diol
5-Nitropyridine-2, 3-diamine (25.0mg, 1.63mmol) was added to diethyl oxalate (5.0mL), and the reaction mixture was stirred at 180 ℃ for 72 hours, then cooled to room temperature. Adding Et2O was added to the reaction mixture to form a solid. The resulting solid was filtered and dried under reduced pressure to give 7-nitropyrido [2,3-b ] as a light brown solid]Pyrazine-2, 3-diol (167.0mg, 49%).
1H-NMR(400MHz，DMSO-d6)；：12.95(s，1H)，12.23(s，1H)，8.93(d，1H，J＝1.8Hz)，8.11(d，1H，J＝1.8Hz)
(c) Synthesis of 2, 3-dichloro-7-nitropyrido [2,3-b ] pyrazine
Reacting 7-nitropyrido [2,3-b ]]Pyrazine-2, 3-diol (167.0mg, 0.80mmol) and POCl3(2.6mL) the mixture was stirred at 150 ℃ for 48 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc ═ 80: 20). Collecting the product-containing fractions, and evaporating to obtain 2, 3-dichloro-7-nitropyrido [2,3-b ] as a white solid]Pyrazine (68.0mg, 35%).
LC/MS ESI(+)：245(M+1)，247(M+3)
(d) Synthesis of tert-butyl (1- (2-chloro-7-nitropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
2, 3-dichloro-7-nitropyrido [2,3-b ] pyrazine (68.0mg, 0.28mmol) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester HCl salt (61.8mg, 0.28mmol) were added to DCM (6.0mL) to which TEA (0.12mL, 0.84mmol) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 70: 30). The product-containing fractions were collected and evaporated to yield the compound tert-butyl (1- (2-chloro-7-nitropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate as a white solid (95.0mg, 87%).
LC/MS ESI(+)：395(M+1)
(e) Synthesis of tert-butyl (1- (2-hydrazino-7-nitropyrido [2,3-b ] pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate
1- (2-chloro-7-nitropyrido [2,3-b ]]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (90.0mg, 0.23mmol) and hydrazine hydrate (24.0. mu.L, 0.68mmol) were dissolved in EtOH (2.3mL), stirred at room temperature for 1 hour, and then concentrated under reduced pressure. Adding Et2O is added to the reaction mixtureIn this, a solid was formed. The resulting solid was filtered and dried under reduced pressure to give (1- (2-hydrazino-7-nitropyridino [2, 3-b) ] a yellow solid compound]Pyrazin-3-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester. Unpurified (1- (2-hydrazino-7-nitropyridino [2, 3-b))]A mixture of t-butyl pyrazin-3-yl) azetidin-3-yl) (methyl) carbamate and trimethyl orthoformate (2.5mL) was stirred at 75 ℃ for 5 hours, then cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered and purified by amine silica gel column chromatography (DCM: MeOH ═ 95: 5). The product-containing fractions were collected and evaporated to give methyl (1- (8-nitropyrido [2, 3-e) as a milky white solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-yl) carbamic acid tert-butyl ester (64.0mg, 66%).
LC/MS ESI(+)：401(M+1)
1H-NMR(400MHz，DMSO-d6)；：10.23(s，1H)，9.47(d，1H，J＝1.8Hz)，9.32(d，1H，J＝1.8Hz)，5.00(m，3H)，4.52(m，1H)，2.95(s，3H)，1.44(s，9H)
(f) Synthesis of N-methyl-1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
Methyl (tert-butyl 1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate (30.0mg, 0.08mmol) was dissolved in DCM (1.0mL), to which TFA (0.5mL) was slowly added at 0 ℃. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 95: 5). The product-containing fractions were collected and evaporated to yield the yellow solid compound N-methyl-1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (20.0mg, 89%).
LC/MS ESI(+)：301(M+1)
1H-NMR(400MHz，DMSO-d6)；：10.22(s，1H)，9.44(d，1H，J＝1.8Hz)，9.30(d，1H，J＝1.8Hz)，4.97(m，1H)，4.53(m，2H)，4.09(m，1H)，3.77(m，1H)，2.32(s，3H)
Example 112
Synthesis of 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-amine
(a) Synthesis of tert-butyl (1- (8-aminopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate
Methyl tert-butyl (1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) carbamate (35.0mg, 0.09mmol) and 10% Pd/C (9.0mg) were dissolved in MeOH (3.0 mL). The flask was replaced with hydrogen, and then stirred at room temperature for 40 minutes. The reaction mixture was filtered through celite, then concentrated under reduced pressure to give tert-butyl (1- (8-aminopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (32.4 mg).
LC/MS ESI(+)：371(M+1)
(b) Synthesis of 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-amine
Unpurified tert-butyl (1- (8-aminopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate (32.4mg, 0.09mmol) was dissolved in DCM (1.5mL) and TFA (0.5mL) was added slowly thereto at room temperature. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (n-Hex: EtOA: MeOH 40:40: 20). The product-containing fractions were collected and evaporated to give the compound 4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-amine as a yellow solid (5.0mg, 21%, 2 steps).
LC/MS ESI(+)：271(M+1)
1H-NMR(400MHz，DMSO-d6)；：9.83(s，1H)，8.05(d，1H，J＝1.8Hz)，7.56(d，1H，J＝1.8Hz)，5.63(s，2H)，4.55(m，2H)，4.11(m，2H)，3.68(m，1H)，2.29(s，3H)
Example 113
Synthesis of N-methyl-1- (8-phenylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine
Pd (PPh)3)4(17.0mg, 0.02mmol) was added to 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol), phenylboronic acid (27.0mg, 0.22mmol), Cs2CO3(122.0mg，0.38mmol)、H2O (0.3mL) and DME (1.2 mL). The reaction mixture was reacted for 30 minutes using a microwave at 50W and 80 ℃ and then cooled to room temperature. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and concentrated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered and dried under reduced pressure to give N-methyl-1- (8-phenylpyrido [2,3-e ] as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) azetidin-3-amine (42.0mg, 85%).
LC/MS ESI(+)：332(M+1)
1H-NMR(400MHz，DMSO-d6)；：10.08(s，1H)，8.91(m，2H)，7.87(m，2H)，7.55(m，2H)，7.44(m，1H)，4.91(m，1H)，4.45(m，2H)，4.02(m，1H)，3.72(m，1H)，2.45(m，1H)，2.31(s，1H)
Example 114
Synthesis of 1- (8- (furan-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
Pd (PPh)3)4(17.0mg, 0.02mmol) was added to 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazole compoundsAnd [4,3-a ]]Pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol), furan-2-ylboronic acid (34.0mg, 0.30mmol), Cs2CO3(122.0mg，0.38mmol)，H2O (0.3mL) and DME (1.2 mL). The reaction mixture was reacted at 50W and 80 ℃ for 30 minutes using microwaves, and then cooled to room temperature. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1:40), fractions containing the product were collected, and concentrated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered and dried under reduced pressure to give 1- (8- (furan-2-yl) pyrido [2, 3-e) as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (31.0mg, 65%).
LC/MS ESI(+)：322(M+1)
1H-NMR(400MHz，DMSO-d6)；：10.07(s，1H)，8.91(s，1H)，8.82(s，1H)，7.87(s，1H)，7.12(m，1H)，6.70(m，1H)，4.91(m，1H)，4.44(m，2H)，4.02(m，1H)，3.72(m，1H)，2.31(s，3H)
Example 115
Synthesis of 1- (2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6-yl) -N-methylazetidin-3-amine
(a) Synthesis of 5-bromo-3- (1H-pyrrol-1-yl) pyridin-2-amine
A mixture of 5-bromopyridine-2, 3-diamine (2000.0mg, 10.6mmol) and 2, 5-dimethoxytetrahydrofuran (1.5mL, 11.7mmol) in AcOH (10.0mL) was stirred at 90 ℃ for 4 hours. The reaction mixture was poured into water, neutralized with 10% aqueous NaOH (pH 7), and then extracted with EtOAc (200.0 mL). The organic layer was washed with brine, anhydrous Na2SO4Dried, filtered and then concentrated. The residue was purified by silica gel column chromatography (EtOAc: n-Hex ═ 1: 4). The product-containing fractions were collected and concentrated to give 5-bromo-3- (1H-pyrrol-1-yl) pyridin-2-amine (614.0mg, 24%) as a yellow solid.
LC/MS ESI(+)：238(M+1)，240(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.13(d，1H，J＝2.1Hz)，7.53(d，1H，J＝2.1Hz)，6.83(m，2H)，6.37(m，2H)，4.63(brs，2H)
(b) Synthesis of 2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6 (5H) -one
A mixture of 5-bromo-3- (1H-pyrrol-1-yl) pyridin-2-amine (200.0mg, 0.84mmol) and triphosgene (374.0mg, 1.26mmol) in dry toluene (8.0mL) was stirred at 110 deg.C for 3 hours, cooled to room temperature, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM: MeOH ═ 30: 1). The product-containing fractions were collected and then concentrated to give the dark green solid compound 2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6 (5H) -one (178.0mg, 80%).
LC/MS ESI(+)：264(M+1)，266(M+3)
1H-NMR(400MHz，DMSO-d6)；：11.86(s，1H)，8.81(m，1H)，8.37(m，1H)，8.28(m，1H)，7.08(m，1H)，6.74(m，1H)
(c) Synthesis of tert-butyl (1- (2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6-yl) azetidin-3-yl) (methyl) carbamate
Reacting 2-bromopyrido [2,3-e ]]Pyrrolo [1,2-a]Pyrazin-6 (5H) -one (178.0mg, 0.67mmol) was dissolved in POCl3(3.0mL) to which DIPEA (141.0. mu.L, 0.81mmol) was then added. The reaction mixture was stirred at 120 ℃ for 12 hours, cooled to room temperature and then concentrated under reduced pressure. Unpurified 2-bromo-6-chloropyrido [2,3-e ]]Pyrrolo [1,2-a]Pyrazine and TEA (0.5mL, 3.35mmol) were dissolved in DMA (7.0mL) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (164.0mg, 0.74mmol) was added thereto. The reaction mixture was stirred at 110 ℃ for 12 hours, cooled to room temperature and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (DCM: MeOH ═ 97: 3). The product-containing fractions were collected and then concentrated to give a brown solid compound (1- (2-bromopyrido [2, 3-)e]Pyrrolo [1,2-a]Pyrazin-6-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (179.0mg, 62%, 2 steps).
LC/MS ESI(+)：432(M+1)，434(M+3)
(d) Synthesis of 1- (2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6-yl) -N-methylazetidin-3-amine
1- (2-bromopyrido [2, 3-e)]Pyrrolo [1,2-a]Pyrazin-6-yl) azetidin-3-yl) (methyl) carbamic acid tert-butyl ester (179.0mg, 0.41mmol) was dissolved in DCM (2.0mL) to which TFA (2.0mL) was added slowly at 0 ℃. The reaction mixture was stirred at room temperature for 2 hours, then concentrated under reduced pressure. By NEt3The residue was neutralized and purified by amine silica gel column chromatography (DCM: MeOH ═ 100:0-90: 10). Collecting the product-containing fractions, and concentrating to obtain 1- (2-bromopyrido [2,3-e ] as brown solid]Pyrrolo [1,2-a]Pyrazin-6-yl) -N-methylazetidin-3-amine (91.0mg, 66%).
LC/MS ESI(+)：332(M+1)，334(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.75(m，1H)，8.39(m，2H)，6.90(m，1H)，6.82(m，1H)，4.54(m，2H)，4.09(m，2H)，3.65(m，1H)，2.28(s，3H)
Example 116
Synthesis of 1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylazetidin-3-amine
(a) Synthesis of 6-bromo-3-nitropyridine-2-amine
2, 6-dibromo-3-nitropyridine (700.0mg, 2.48mmol) was added to a 2M solution of ammonia in EtOH (25.0mL, 49.66 mmol). The mixture was stirred at room temperature for 12 hours, then concentrated under reduced pressure to give 6-bromo-3-nitropyridin-2-amine (526.0mg, 97%) as a yellow solid.
1H-NMR(400MHz，DMSO-d6)；：8.26(m，3H)，6.91(d，1H，J＝8.4Hz)
(b) Synthesis of 6-bromopyridine-2, 3-diamine
6-bromo-3-nitropyridin-2-amine (520.0mg, 2.39mmol) and SnCl22H2O (2140.0mg, 9.54mmol) was added to DMF (8.6mL) and the reaction mixture was stirred at 80 ℃ for 2 h. The reaction mixture was cooled to room temperature, basified with 1N NaOH (pH 9), and then extracted with EtOAc. With anhydrous Na2SO4The organic layer was dried, filtered, and then concentrated under reduced pressure to give 6-bromopyridine-2, 3-diamine (238.0mg, 44%) as a brown solid.
1H-NMR(400MHz，DMSO-d6)；：6.62(d，1H，J＝8.4Hz)，6.48(d，1H，J＝8.4Hz)，5.81(brs，2H)，4.79(brs，2H)
(c) Synthesis of 6-bromopyrido [2,3-b ] pyrazine-2, 3-diol
6-bromopyridine-2, 3-diamine (238.0mg, 1.26mmol) was added to diethyl oxalate (4.0 mL). The mixture was stirred at 150 ℃ for 4 hours and then cooled to room temperature. Et was added2O to form a solid. The solid formed was filtered and then dried under reduced pressure to give a brown solid compound, 6-bromopyrido [2,3-b]Pyrazine-2, 3-diol (250.0mg, 82%).
1H-NMR(400MHz，DMSO-d6)；：12.50(brs，1H)，12.06(brs，1H)，7.36(m，2H)
(d) Synthesis of 2,3, 6-trichloropyrido [2,3-b ] pyrazine
Reacting 6-bromopyrido [2,3-b ]]Pyrazine-2, 3-diol (250.0mg, 1.03mmol) and POCl3(4.0mL) of the mixture was stirred at 150 ℃ for 10 hours and then cooled to room temperature. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 70: 30). Collecting the product-containing fractions, and concentrating to obtain white solid compound 2,3, 6-trichloropyrido [2,3-b ]]Pyrazine (180.0mg, 63%).
LC/MS ESI(+)：234(M+1)，236(M+3)
1H-NMR(400MHz，DMSO-d6)；：8.65(d，1H，J＝8.8Hz)，8.08(d，1H，J＝8.8Hz)
(e) Synthesis of 2, 6-dichloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Hydrazine monohydrate (18.0. mu.L, 0.51mmol) was added to 2,3, 6-trichloropyrido [2,3-b ]]Pyrazine (80.0mg, 0.34mmol) and EtOH (1.4 mL). The reaction mixture was stirred at room temperature for 90 minutes. Et was added thereto2O to form a solid. The solid formed was filtered and dried under reduced pressure to give 2, 6-dichloro-3-hydrazinopyrido [2,3-b ] as a yellow solid]A pyrazine. Unpurified 2, 6-dichloro-3-hydrazinopyrido [2,3-b ]]Pyrazine (60.0mg, 0.26mmol) was dissolved in trimethyl orthoformate (3.0mL) and stirred at 85 ℃ for 12 h. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 70: 30). Collecting the product-containing fractions, and concentrating to obtain light brown solid compound 2, 6-dichloropyrido [3,2-e][1,2,4]Triazolo [4,3-a]Pyrazine (35.0mg, 56%, 2 steps).
1H-NMR(400MHz，DMSO-d6)；：10.14(s，1H)，8.60(d，1H，J＝8.4Hz)，7.96(d，1H，J＝8.4Hz)
(f) Synthesis of 1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylazetidin-3-amine
2, 6-dichloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (35.0mg, 0.12mmol) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (41.0mg, 0.18mmol) were added to DCM (1.0mL), to which TEA (51.0. mu.L, 0.37mmol) was slowly added at room temperature. The reaction mixture was stirred at room temperature for 30 minutes, then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 70: 30). The product-containing fractions were collected and concentrated to give the compound tert-butyl (1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) azetidin-3-yl) (methyl) carbamate as a yellow solid. To a mixture of tert-butyl (1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) azetidin-3-yl) (methyl) carbamate and DCM (1.0mL) was added TFA (0.4mL), and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and concentrated to give the light brown solid compound 1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylazetidin-3-amine (30.0mg, 86%, 2 steps).
LC/MS ESI(+)：290(M+1)
1H-NMR(400MHz，DMSO-d6)；：9.31(s，1H)，8.02(d，1H，J＝8.4Hz)，7.61(d，1H，J＝8.4Hz)，4.90(m，1H)，4.43(m，2H)，4.00(m，1H)，3.73(m，1H)，2.30(s，3H)
Example 117
Synthesis of 1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
(a) Synthesis of 6-bromopyridine-3, 4-diamine
2-bromo-5-nitropyridin-4-amine (300.0mg, 2.48mmol) and Fe (300.0mg, 5.37mmol) were added to AcOH and the reaction mixture was stirred at 75 ℃ for 4h and then cooled to room temperature. The reaction mixture was filtered through celite and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (n-Hex: EtOAc 50: 50). The product-containing fractions were collected and then concentrated to give 6-bromopyridine-3, 4-diamine (200.0mg, 78%) as a brown solid.
1H-NMR(400MHz，DMSO-d6)；：7.39(s，1H)，6.42(s，1H)，5.74(brs，2H)，4.66(brs，2H)
(b) Synthesis of 7-bromopyrido [3,4-b ] pyrazine-2, 3-diol
6-bromopyridine-3, 4-diamine (200.0mg, 1.06mmol) was added to oxalic acidDiethyl ester (3.0 mL). The mixture was stirred at 130 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid. The resulting solid was filtered and dried under reduced pressure to give 7-bromopyrido [3,4-b ] as a brown solid]Pyrazine-2, 3-diol (195.0mg, 76%).
1H-NMR(400MHz，DMSO-d6)；：12.23(brs，1H)，12.11(brs，1H)，8.09(s，1H)，7.05(s，1H)
(c) Synthesis of 2,3, 7-trichloropyrido [3,4-b ] pyrazine
Reacting 7-bromopyrido [3,4-b ]]Pyrazine-2, 3-diol (190.0mg, 0.79mmol) and POCl3(3.0mL) of the mixture was stirred at 150 ℃ for 12 hours and then cooled to room temperature. Et was added thereto2O to form a solid and filtering the formed solid. The resulting solid was purified by silica gel column chromatography (n-Hex: EtOAc ═ 50: 50). Collecting the product-containing fractions, and concentrating to obtain white solid compound 2,3, 7-trichloropyrido [3,4-b ]]Pyrazine (1.0mg, 5%).
LC/MS ESI(+)：234(M+1)，236(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.39(s，1H)，8.31(s，1H)
(d) Synthesis of 4, 8-dichloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazine
Hydrazine monohydrate (1.9. mu.L, 0.05mmol) was added to 2,3, 7-trichloropyrido [3,4-b ] at 0 deg.C]Pyrazine (10.0mg, 0.03mmol) and EtOH (0.5 mL). The reaction mixture was stirred at room temperature for 3 hours. Et was added thereto2O to form a solid. Filtering the solid formed, drying under reduced pressure to obtain the compound 3, 7-dichloro-2-hydrazinopyrido [3,4-b ] as a red solid]A pyrazine. Unpurified 3, 7-dichloro-2-hydrazinopyrido [3,4-b ]]Pyrazine was dissolved in trimethyl orthoformate (0.3mL) and the reaction mixture was stirred at 80 ℃ for 5 hours. The reaction mixture was cooled to room temperature. Et was added thereto2O to form a solid. The solid formed was filtered and dried under reduced pressure to form 4, 8-dichloropyrazole as an orange solidPyrido [3,4-e][1,2,4]Triazolo [4,3-a]Pyrazine (3.5mg, 42%, 2 steps).
LC/MS ESI(+)：240(M+1)
1H-NMR(400MHz，CDCl3)；：9.35(s，1H)，9.19(s，1H)，7.90(s，1H)
(e) Synthesis of 1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine
4, 8-dichloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazine (3.5mg, 0.01mmol) and azetidin-3-yl (methyl) carbamic acid tert-butyl ester hydrochloride (4.1mg, 0.02mmol) were added to DCM (0.2mL), and TEA (5.0. mu.L, 0.04mmol) was added thereto at room temperature. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure to give tert-butyl (1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate. To a mixture of unpurified tert-butyl (1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) (methyl) carbamate and DCM (1.0mL) was added TFA (0.2mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and concentrated to give the light brown solid compound 1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (2.0mg, 57%, 2 steps).
LC/MS ESI(+)：290(M+1)
1H-NMR(400MHz，CDCl3)；：9.12(s，1H)，8.77(s，1H)，7.62(s，1H)，5.07(m，1H)，4.59(m，2H)，4.18(m，1H)，3.88(m，1H)，2.50(s，3H)
Example 118
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine methanesulfonate
Methanesulfonic acid (38.9 μ L, 0.60mmol) was slowly added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (200.0mg, 0.60mmol) and ethanol (3.0 mL). The reaction mixture was stirred at room temperature for 20 hours to form a solid. The solid formed was filtered under reduced pressure, washed with ethanol and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine mesylate, a cream-white solid (226.0mg, 88%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.02(s，1H)，9.14(brs，2H)，8.99(d，1H，J＝2.3Hz)，8.67(d，1H，J＝2.3Hz)，5.03(m，1H)，4.84(m，1H)，4.58(m，1H)，4.40(m，1H)，4.27(m，1H)，2.68(s，3H)，2.30(s，3H)
Example 119
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine maleate
Maleic acid (139.0, 1.20mmol) was added slowly to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (200.0mg, 0.60mmol) and ethanol (3.0 mL). The reaction mixture was stirred at room temperature for 20 hours. The solid formed was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine maleate as a white solid (242.0mg, 90%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.02(s，1H)，8.99(brs，2H)，8.98(d，1H，J＝2.3Hz)，8.66(d，1H，J＝2.3Hz)，6.02(s，2H)，5.03(m，1H)，4.82(m，1H)，4.58(m，1H)，4.38(m，1H)，4.23(m，1H)，2.66(s，3H)
Example 120
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2-hydroxypropane-1, 2, 3-tricarboxylate
2-hydroxypropane-1, 2, 3-tricarboxylic acid (230.0mg, 1.20mmol) was added slowly to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (200.0mg, 0.60mmol) and THF (6.0 mL). The reaction mixture was stirred at room temperature for 20 hours. The solid formed was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2-hydroxypropane-1, 2, 3-tricarboxylate (246.0mg, 78%) as a white solid.
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.98(s，1H)，8.94(d，1H，J＝2.3Hz)，8.63(d，1H，J＝2.3Hz)，4.96(m，1H)，4.62(m，1H)，4.50(m，1H)，4.18(m，1H)，3.95(m，1H)，2.55(m，4H)，2.47(s，3H)
Example 121
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine nitrate
70% nitric acid (13.0. mu.L, 0.30mmol) was added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.5 mL). The reaction mixture was stirred at room temperature for 12 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine nitrate as a white solid (55.8mg, 94%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.02(s，1H)，9.15(brs，2H)，8.98(d，1H，J＝2.4Hz)，8.66(d，1H，J＝2.4Hz)，5.03(m，1H)，4.83(m，1H)，4.58(m，1H)，4.40(m，1H)，4.27(m，1H)，2.69(s，3H)
Example 122
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydroiodide
55% HI (41.0. mu.L, 0.30mmol) was added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.5 mL). The reaction mixture was stirred at room temperature for 12 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydroiodide (60.0mg, 87%) as a yellow solid.
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.02(s，1H)，9.11(brs，2H)，8.99(d，1H，J＝2.0Hz)，8.67(d，1H，J＝2.0Hz)，5.03(m，1H)，4.83(m，1H)，4.58(m，1H)，4.40(m，1H)，4.26(m，1H)，2.69(s，3H)
Example 123
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine phosphate
85% phosphoric acid (25.9. mu.L, 0.22mmol) was added slowly to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (2.0 mL). The reaction mixture was stirred at room temperature for 20 hours. The solid formed was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine phosphate (59.0mg, 91%) as a white solid.
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.97(s，1H)，8.93(d，1H，J＝2.3Hz)，8.61(d，1H，J＝2.3Hz)，4.94(m，1H)，4.59(m，1H)，4.48(m，1H)，4.15(m，1H)，3.90(m，1H)，2.42(s，3H)
Example 124
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 4,4' -methylenebis (3-hydroxy-2-naphthoic acid) salt
Pamoic acid (87.2mg, 0.22mmol) was added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (2.0 mL). The reaction mixture was stirred at room temperature for 20 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and then dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 4,4' -methylenebis (3-hydroxy-2-naphthoic acid) salt as a green solid (95.0mg, 88%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.98(s，1H)，8.91(d，1H，J＝2.3Hz)，8.63(d，1H，J＝2.3Hz)，8.35(s，2H)，8.14(m，2H)，7.79(m，2H)，7.30(m，2H)，7.15(m，2H)，5.04(m，1H)，4.91(m，1H)，4.74(s，2H)，4.59(m，1H)，4.47(m，1H)，4.29(m，1H)，2.72(s，3H)
Example 125
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrobromide
A solution of 33% hydrogen bromide in acetic acid (41.3. mu.L, 0.23mmol) was added slowly to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.5 mL). The reaction mixture was stirred at room temperature for 20 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrobromide as a white solid (54.3mg, 87%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.04(s，1H)，9.23(brs，2H)，9.00(d，1H，J＝2.2Hz)，8.67(d，1H，J＝2.2Hz)，5.05(m，1H)，4.87(m，1H)，4.59(m，1H)，4.43(m，1H)，4.29(m，1H)，2.69(s，3H)
Example 126
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine sulfate
A solution of sulfuric acid (22.5mg, 0.23mmol) in ethanol (0.5mL) was added slowly to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.0 mL). The reaction mixture was stirred at room temperature for 20 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine sulfate as a white solid (64.6mg, 99%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.03(s，1H)，9.20(brs，2H)，9.01(d，1H，J＝2.2Hz)，8.67(d，1H，J＝2.2Hz)，5.05(m，1H)，4.86(m，1H)，4.60(m，1H)，4.43(m，1H)，4.29(m，1H)，2.69(t，3H，J＝5.0Hz)
Example 127
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (2R,3R) -2, 3-dihydroxysuccinate salt
A solution of (2R,3R) -2, 3-dihydroxysuccinic acid (45mg, 0.30mmol) in ethanol (0.5mL) was slowly added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.0 mL). The reaction mixture was stirred at room temperature for 48 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (2R,3R) -2, 3-dihydroxysuccinate as a white solid (62.9mg, 86%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.97(s，1H)，8.92(d，1H，J＝2.2Hz)，8.60(d，1H，J＝2.2Hz)，4.94(m，1H)，4.57(m，1H)，4.47(m，1H)，4.14(m，1H)，4.13(s，2H)，3.90(m，1H)，2.42(s，3H)
Example 128
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (1S) - (+) -10-camphorsulfonate
(1S) - (+) -10-Camphorsulfonic acid (69.5mg, 0.30mmol) was added to a suspension of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (1.5 mL). The reaction mixture was stirred at room temperature for 12 hours. The resulting solid was filtered under reduced pressure, washed with ethanol, and dried under reduced pressure to give 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (1S) - (+) -10-camphorsulfonate (39.0mg, 46%) as a white solid.
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.03(s，1H)，9.03(brs，2H)，8.99(d，1H，J＝2.0Hz)，8.67(d，1H，J＝2.0Hz)，5.04(m，1H)，4.84(m，1H)，4.59(m，1H)，4.40(m，1H)，4.25(m，1H)，2.86(d，1H，J＝14.8Hz)，2.71(m，1H)，2.68(S，3H)，2.36(d，1H，J＝14.8Hz)，2.23(m，1H)，1.93(m，1H)，1.85(1H，m)，1.80(d，1H，J＝18.4Hz)，1.27(m，2H)，1.05(S，3H)，0.75(S，3H)
Example 129
Synthesis of 8-bromo-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine
(a) Synthesis of 7-bromo-2-chloro-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-b ] pyrazin-3-amine
TEA (150.0. mu.L, 1.07mmol) was added to a mixture of 7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (100.0mg, 0.36mmol), 1-methylpyrrolidin-3-amine (43.0mg, 0.43mmol) and DCM (1.8mL) and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and concentrated to give the compound 7-bromo-2-chloro-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-b ] pyrazin-3-amine (30.0mg, 24%) as a yellow solid.
LC/MS ESI(+)：342(M+1)，344(M+3)
(b) Synthesis of 8-bromo-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine
Hydrazine monohydrate (5.5. mu.L, 0.18mmol) was added to 7-bromo-2-chloro-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-b ]]Pyrazin-3-amine (26.0mg, 0.06mmol) and EtOH (1.0mL) were suspended. The reaction mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was dissolved in trimethyl orthoformate (1.0mL) and the reaction mixture was stirred at 100 ℃ for 5 hours. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 1: 40). The product-containing fractions were collected and concentrated. Adding Et2O was added to the residue to form a solid. The resulting solid was filtered and dried under reduced pressure to give 8-bromo-N- (1-methylpyrrolidin-3-yl) pyrido [2,3-e as a yellow solid][1,2,4]Triazolo [4,3-a]Pyrazin-4-amine (6.0mg, 30%).
LC/MS ESI(+)：348(M+1)，350(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.95(s，1H)，8.92(d，1H，J＝2.0Hz)，8.78(m，1H)，8.61(d，1H，J＝2.0Hz)，4.72(m，1H)，2.86(m，1H)，2.68-2.50(m，3H)，2.33-2.10(m，4H)，2.01(m，1H)
Example 130
Synthesis of 1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (S) -2-hydroxypropionate
A solution of (S) -2-hydroxypropionic acid in water (0.8mL) (135.0mg, 1.50mmol) was added to 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (50.0mg, 0.15mmol) and ethanol (0.8mL) in suspension. The reaction mixture was stirred at room temperature for 3 hours, then concentrated under reduced pressure. Reaction of EtOH and Et2O was added to the residue to form a solid. Filtering to obtainTo solid by Et2Washing with O, and drying under reduced pressure to obtain yellow solid compound 1- (8-bromopyrido [2,3-e ]][1,2,4]Triazolo [4,3-a]Pyrazin-4-yl) -N-methylazetidin-3-amine (S) -2-hydroxypropionate (34.0mg, 53%).
LC/MS ESI(+)：334(M+1)，336(M+3)
1H-NMR(400MHz，DMSO-d6)；：9.96(s，1H)，8.91(d，1H，J＝2.0Hz)，8.60(d，1H，J＝2.0Hz)，4.91(m，1H)，4.45(m，2H)，4.02(m，2H)，3.75(m，1H)，2.33(s，3H)，1.22(m，3H)
Example 131
Synthesis of N- (azetidin-3-ylmethyl) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine trifluoroacetate
(a) Synthesis of tert-butyl 3- (((7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) amino) methyl) azetidine-1-carboxylate
7-bromo-2, 3-dichloropyrido [2,3-b ] pyrazine (100.0mg, 0.56mmol) was dissolved in DCM (5.6mL) and tert-butyl 3- (aminomethyl) azetidine-1-carboxylate (104.1mg, 0.56mmol) and TEA (0.23mL, 1.68mmol) were added slowly thereto at 0 ℃. The reaction mixture was stirred at room temperature for 48 hours, then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-Hex: EtOAc 70: 30). The product-containing fractions were collected and concentrated to give the compound tert-butyl 3- (((7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) amino) methyl) azetidine-1-carboxylate (72.0mg, 30%) as a yellow solid.
LC/MS ESI(+)：428(M+1)
1H-NMR(400MHz，DMSO-d6)；：8.85(d，1H，J＝2.4Hz)，8.48(d，1H，J＝2.4Hz)，8.20(t，1H，J＝5.6Hz)，3.90(m，2H)，3.70(m，4H)，2.92(m，1H)，1.38(s，9H)
(b) Synthesis of tert-butyl 3- (((7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) amino) methyl) azetidine-1-carboxylate
Reacting 3- (((7-bromo-2-chloropyrido [2, 3-b))]Pyrazin-3-yl) amino) methyl) azetidine-1-carboxylic acid tert-butyl ester (50.0mg, 0.12mmol) and hydrazine monohydrate (12.0 μ L, 0.35mmol) were dissolved in EtOH (0.6mL) and the reaction mixture was stirred at room temperature for 1 hour. Et was added thereto2O to form a solid. The resulting solid was filtered and dried under reduced pressure to give 3- (((7-bromo-2-hydrazinopyrido [2, 3-b)) as a yellow solid]Pyrazin-3-yl) amino) methyl) azetidine-1-carboxylic acid tert-butyl ester (48.0mg, 98%).
LC/MS ESI(+)：424(M+1)
(c) Synthesis of N- (azetidin-3-ylmethyl) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine trifluoroacetate
The reaction mixture of tert-butyl 3- (((7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) amino) methyl) azetidine-1-carboxylate (50.0mg, 0.12mmol) and trimethyl orthoformate (1.5mL) was stirred at 70 ℃ for 12 hours, cooled to room temperature, and then dried under reduced pressure to give tert-butyl 3- (((8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) methyl) azetidine-1-carboxylate. TFA (0.3mL) was added to a mixture of unpurified tert-butyl 3- (((8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) methyl) azetidine-1-carboxylate and DCM (1.0mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was purified by amine silica gel column chromatography (MeOH: DCM ═ 5: 95). The product-containing fractions were collected and concentrated to give the light brown solid compound N- (azetidin-3-ylmethyl) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-amine trifluoroacetate (1.5mg, 3%, 2 steps).
LC/MS ESI(+)：334(M+1)
1H-NMR(400MHz，MeOH-d4)；：9.77(s，1H)，8.80(d，1H，J＝2.0Hz)，8.64(m，1H)，3.94(m，2H)，3.76(m，2H)，3.63(m，2H)，3.35(s，1H)
Example 132
Synthesis of 4- (azetidin-3-ylmethoxy) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine hydrochloride
(a) Synthesis of tert-butyl 3- (((7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) oxy) methyl) azetidine-1-carboxylate
60% NaH (10mg, 0.25mmol) was added to a solution of tert-butyl 3- (hydroxymethyl) azetidine-1-carboxylate (47.9mg, 0.25mmol) in THF (2.5mL) at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 30 minutes, and then 7-bromo-2, 3-dichloropyrido [2,3-b ] was added thereto at room temperature]Pyrazine (60.0mg, 0.25 mmol). The reaction mixture was stirred at rt for 2h, diluted with EtOAc. With anhydrous MgSO4Dried, filtered and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (n-Hex: EtOAc ═ 2: 1). Collecting the product-containing fractions, and concentrating to obtain white solid compound 3- (((7-bromo-2-chloropyrido [2, 3-b))]Pyrazin-3-yl) oxy) methyl) azetidine-1-carboxylic acid tert-butyl ester (65.8mg, 61%).
LC/MS ESI(+)：429(M+1)，431(M+3)，451(M+23)，453(M+25)
1H-NMR(400MHz，CDCl3)；：9.00(d，1H，J＝2.3Hz)，8.45(d，1H，J＝2.3Hz)，4.79(d，2H，J＝6.6Hz)，4.13(t，2H，J＝8.5Hz)，3.85(dd，2H，J＝5.2，3.6Hz)，3.13(m，1H)，1.45(s，9H)
(b) Synthesis of tert-butyl 3- (((8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) oxy) methyl) azetidine-1-carboxylate
Hydrazine monohydrate (14.1 μ L, 0.45mmol) was added to a mixture of tert-butyl 3- (((7-bromo-2-chloropyrido [2,3-b ] pyrazin-3-yl) oxy) methyl) azetidine-1-carboxylate (63.8mg, 0.15mmol) and EtOH (1.5mL) and the reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure and unpurified tert-butyl 3- (((7-bromo-2-hydrazinopyrido [2,3-b ] pyrazin-3-yl) oxy) methyl) azetidine-1-carboxylate was dissolved in trimethyl orthoformate (1.5 mL). The reaction mixture was stirred at 80 ℃ for 4 hours and then concentrated under reduced pressure to give the unpurified brown solid compound, tert-butyl 3- (((8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) oxy) methyl) azetidine-1-carboxylate.
LC/MS ESI(+)：435(M+1)，437(M+3)，457(M+23)，459(M+25)
(c) Synthesis of 4- (azetidin-3-ylmethoxy) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine hydrochloride
Unpurified tert-butyl 3- (((8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) oxy) methyl) azetidine-1-carboxylate was dissolved in DCM (1.5mL) and a solution of 4N HCl in 1, 4-dioxane (75.0 μ L, 0.30mmol) was slowly added thereto at room temperature. The mixture was stirred at room temperature for 2 hours, then concentrated under reduced pressure. The residue was dissolved in DMSO (0.5mL) and then purified by preparative HPLC (YL9110S young lin, acetonitrile: water 8: 92). The product-containing fractions were collected and lyophilized to give the compound 4- (azetidin-3-ylmethoxy) -8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine hydrochloride as a white solid (6.4mg, 11%, 3 steps).
LC/MS ESI(+)：335(M+1)，337(M+3)
1H-NMR(400MHz，DMSO-d6)；：10.05(s，1H)，9.15(d，1H，J＝2.0Hz)，8.97(brs，2H)，8.78(d，1H，J＝2.0Hz)，4.79(d，2H，J＝6.64Hz)，4.04(m，2H)，3.90(m，2H)，3.33(m，1H)
Experimental example 1: binding affinity assay for human histamine 4 receptor (hH4R)
Each of the compounds of the present invention prepared in examples was diluted 1000-fold (v/w) with DMSO, and then 1. mu.L of the diluted compound solution was mixed with 99. mu.LL of assay buffer (50mM tris-HCl pH7.4, 5mM EDTA) were mixed to give concentrations of 0.1, 0.3, 1,3, 10 and 100. mu.M. mu.L of the prepared compound solution was transferred to each well of a 96-well plate, and then 20. mu.L of 100. mu.M histamine diluted with assay buffer solution and 1% DMSO was transferred to each well to calculate the extent of nonspecific binding and total binding. Mu.g of human histamine 4 receptor-overexpressed cell membranes (Perkinelmer) were diluted into 160. mu.L of assay buffer and transferred to each well. Will 23H]Labeled histamine (Perkinelmer) was diluted to a concentration of 1. mu.M, and 20. mu.L was dispensed in each well, which was then kept in an incubator at 27 ℃ for 30 minutes. After the reaction, 200. mu.L of the mixture was transferred to a glass fiber plate, which had been preimpregnated with 0.5% of polyvinylamine, and then non-specific binding was removed in vacuo3H]Labeled Histamine after washing 4 times with 200. mu.L of a washing buffer solution (50mM tris-HCl, pH7.4), the plate was oven-dried at 37 ℃ for 18 hours, 100. mu.L of β scintillation cocktail solution was added to each well, and after 1 hour, the value was determined by using Wallac β -counter TriLux [ ]3H]CPM (counts/min) value of labeled histamine. The binding affinity of the histamine 4 receptor of the present inventors (IC50) was analyzed using the Excel program, and the analytical structure is shown in table 1 below.
[ Table 1]
Experimental example 2: binding affinity assay for human histamine 3 receptor (hH3R)
Each of the compounds of the invention prepared in the examples was prepared in DMSO at concentrations of 0.02, 0.06, 0.3 and 2mMmu.L of the prepared compound solution was dispensed into each well of a 96-well plate, and then 10. mu.L of 200. mu.M (R) (-) - α -methyl histamine (RaMH) diluted with assay buffer and 1% DMSO was transferred into each well to calculate the degree of nonspecific binding and total binding, 180. mu.L of assay buffer (50mM tris-HCl pH7.4, 5mM MgCl. sub.2) 15 μ g of human histamine 3 receptor-overexpressing cell membrane (Perkinelmer) was diluted and transferred to each well. Will 23H]Labeled N- α -Methylhistamine (Perkinelmer) was diluted to a concentration of 20. mu.M, 10. mu.L was added to each well, which was then kept in an incubator at 27 ℃ for 30 minutes after the reaction, 200. mu.L of the mixture was transferred to a glass fiber plate, which was pre-impregnated with 0.5% polyvinylamine, followed by vacuum removal of non-specific binding [ ] [, ]3H]Labeled N- α -Methylhistamine after washing 5 times with 200. mu.L of a washing buffer solution (50mM tris-HCl, pH7.4), the plate was oven-dried at 37 ℃ for 18 hours, 100. mu.L of a β scintillation cocktail solution was added to each well, and after 1 hour, the concentration was measured by using Wallac β -counter TriLux3H]Cpm of labeled N- α -methyl histamine the binding affinity of the histamine 3 receptor of the present inventors was analyzed using the Excel program (IC50), and the analytical structure is shown in table 2 below.
[ Table 2]
Experimental example 3: binding affinity assay for human 5-hydroxytryptamine 3 receptor (human 5-HT3 receptor)
Binding assays for the 5-hydroxytryptamine 3 receptor of the present inventors were performed in Cerep (Poitiers, France) and the analytical structures are shown in Table 3 below.
[ Table 3]
| Compound (I) | Binding affinity (IC50) of human 5-hydroxytryptamine 3 receptor (h5-HT3R) |
| Compound 18 | 2.9μM |
| Compound 24 | 9.1μM |
| Compound 25 | >10μM(0％*) |
| Compound 48 | >10μM(28％*) |
| Compound 93 | >10μM(18％*) |
| Compound 95 | >10μM(25％*) |
*: indicates the degree of% inhibition at 10. mu.M
Experimental example 4: solubility testing in artificial gastrointestinal solutions
[ first solution ]
2.0g of sodium chloride was added to 7.0mL of 37% hydrochloric acid and water to make the total volume 1L. The pH was adjusted to exactly 1.2 with 1N sodium hydroxide or 1N hydrochloric acid. A small amount (20mL) of this solution was taken and 25. mu.L of triton X-100 was added to make the total volume 25 mL.
[ second solution ]
0.348g of sodium hydroxide and 3.438g of NaH2PO46.186g of sodium chloride and water were added to make the total volume 1L. The pH was adjusted to exactly 6.5 with 1N sodium hydroxide or 1N hydrochloric acid. A small amount (20mL) of this solution was taken and 41.25mg of hydrated sodium taurocholate dissolved in ethanol and 25. mu.L of 750mM lecithin were added to make the total volume 25 mL.
[ experiment ]
For the compounds of the present invention prepared in the examples, test samples were prepared with the first solution and the second solution each having a concentration of 2mg/mL, and then they were stirred for 1 minute, sonicated for 1 minute, heated at 37 ℃ for 1 hour for the first solution, and heated at 37 ℃ for 2 hours for the second solution. The heated test sample solution is filtered to remove undissolved compounds therein. Finally, 100. mu.L of a completely dissolved solution sample of the compound was collected, and 100. mu.L of CH was added thereto3CN to make test sample solution. For each analysis of the test sample solution, the solubility was determined by experimental liquid chromatography. The results of the solubility tests in the first solution and the second solution are shown in table 4 below.
[ Table 4]
Experimental example 5: metabolic stability test
[ preparation of microsomal solution ]
Microsomes (human, mouse, rat) having a protein concentration of 20mg/mL were diluted with 0.1M phosphate buffer (pH: 7.4) to a concentration of 1.316 mg/mL. The compound of the present invention prepared in example was dissolved in DMSO to prepare a 2.5mM solution, which was diluted with distilled water to prepare a 125. mu.M solution. The microsomal diluted solution and the compound diluted solution prepared as described above were mixed to prepare a microsomal solution having a protein concentration of 1.25mg/mL and a compound concentration of 6.25. mu.M.
[ preparation of NADPH solution ]
NADPH was dissolved in 0.1M phosphate buffer to make a 5mM solution.
[ experiment ]
Each 80. mu.L of the 6.25. mu.M concentration microsomal solutions of the compound of the present invention prepared in the examples was placed in a bath at 37 ℃ and maintained for 5 minutes, and then the reaction was initiated by making 1mg/mL microsomal protein, 5. mu.M compound and 1mM NADPH final concentration by adding 20. mu.L of 5mM NADPH. The reaction was stopped by adding 100 μ L acetonitrile at 0, 10, 20 and 30 minutes, centrifuged, and the supernatant was analyzed by using liquid chromatography. Half-lives were determined using the retained compound peak areas at 0, 10, 20 and 30 minutes, from which CL indicative of metabolic stability was calculatedh.intAnd (4) parameters. The metabolic stability results are shown in table 5 below.
[ Table 5]
NCy: not calculable (99.8% retained in 30 minutes)
Experimental example 6: pharmacokinetic testing
[ preparation of drug ]
The compound of the present invention prepared in example was dissolved in 20% hydroxypropyl- β -cyclodextrin solution to make 5mg/mL administration solution for mouse, and DMSO (5%), 5% glucose solution (94.75%), and 2N hydrochloric acid (0.25%) were added in order to make 2mg/mL administration solution for rat.
[ experiment ]
The weight of each of ICR mice and SD rats is suitably 20-30 g and 200-300 g, and the dose of the compound of the present invention is 10mL/kg for mice and 5mL/kg for rats, administered orally using Zonde. Blood was collected at 0.25, 0.5, 1,2,4, 6, 8 and 24 hours by taking a sample of blood using a capillary orbital vein coated with an anticoagulant, and then plasma was separated using a centrifuge tube and kept in a refrigerated storage.
[ analysis ]
Plasma and standard concentration material collected from animals were pretreated using solid phase extraction and the concentration of the compounds of the invention was determined using a liquid chromatography mass spectrometer (Agilent HPLC, API-3000).
From the values obtained, the pharmacokinetic parameters, half-life (t) were determined using WinNonlin (version 6.2)1/2) Maximum blood concentration (C)max) And Area Under Curve (AUC)inf) As shown in tables 6 and 7 below.
[ Table 6]
[ Table 7]
Experimental example 7: histamine-induced mast cell migration
[ animals ]
Female Balb/c mice (7 weeks old, 20. + -.3 g) were purchased from OrientBio Co., Ltd. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment ]
Mice were given either vehicle (20% cyclodextrin in double distilled water) or compound by oral administration. The compound (20mg/kg) was dissolved in vehicle and administered in a volume of 10 mL/kg. After 15 minutes, mice were challenged with PBS (phosphate buffered saline) or 0.1M histamine (dissolved in PBS) aerosol inhalation for 20 min. This process was repeated for 2 days. After the final challenge, the trachea was cleared of blood by perfusion with PBS, extracted, and fixed with 10% (w/v) formaldehyde for subsequent paraffin cross-sections and toluidine blue staining. Total mast cells were calculated and reported as The mean (The Journal of Pharmacology and Experimental Therapeutics, 2004, 309, 404-. The inhibitory effect of the compounds is shown in table 8. Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant.
[ Table 8]
"1" refers to statistical significance.
Experimental example 8: model of itch in histamine-induced ICR mice
[ animals ]
Female ICR mice (8 weeks old) were purchased from OrientBio co., Ltd. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment: causes of pruritus and determination thereof
24 hours prior to this experiment, hair was cut using a clipper (8000AD, THRIVE) in the back mouth under isoflurane anesthesia. Histamine (300nmol) was dissolved in saline and injected intradermally into the posterior mouth portion in a volume of 40 μ L. Mice were given either vehicle (20% cyclodextrin in double distilled water) or compound by oral administration. The compound (50mg/kg) was dissolved in vehicle (20% cyclodextrin in double distilled water) and administered orally at 10 mL/kg. Mice (n 10/group) were divided into 3 groups (normal group, control group, and experimental group). Animals were randomly grouped according to similar weight distribution. The vehicle or compound was administered 20 minutes before injection of histamine or saline, and the animals were returned to an acrylic cage consisting of 4 chambers (about 30x30x30cm, produced ad libitum) immediately after transdermal injection, allowed to acclimate for 1 hour prior to the experiment, and observed for a pruritic response. A camera (Samsung, VLUUNV30) was positioned in the upper part of the mice to record their response. Pruritus was determined by blind counting of scratching times over a 20min period immediately following intradermal injection. Scratching times were defined as 3 or more separate rapid scratching movements around the injection site using the hind paw (j. allergy clin. immunol.2007, 19(1), 176-. All data were analyzed by using Excel and Prism, and all values were expressed as mean ± s.e.m. The inhibitory effect of the compound is shown as a percentage of the maximal response to histamine (control 100%).
Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant. The inhibitory effect of the compounds is shown in table 9 below.
[ Table 9]
| Compound (I) | Number of scratching (% change compared to control) |
| Compound 4 | 94.31 |
| Compound 7 | 61.21 |
| Compound 18 | 84.31 |
| Compound 24 | 93.61 |
| Compound 31 | 89.31 |
| Compound 48 | 67.31 |
| Compound 95 | 98.21 |
"1" refers to statistical significance.
Experimental example 9: substance P-induced pruritus model in ICR mice
[ animals ]
Female ICR mice (8 weeks old) were purchased from OrientBio co., Ltd. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment: causes of pruritus and determination thereof
24 hours prior to this experiment, hair was cut using a clipper (8000AD, THRIVE) in the back mouth under isoflurane anesthesia. Substance P (100nmol) was dissolved in saline and a volume of 40. mu.L was injected intradermally into the posterior mouth. Mice were given either vehicle (20% cyclodextrin in double distilled water) or compound by oral administration. The compound (50mg/kg) was dissolved in vehicle (20% cyclodextrin in double distilled water) and administered orally in a volume of 10 mL/kg.
Mice (n 10/group) were divided into 3 groups (normal group, control group, and experimental group). Animals were randomly grouped according to similar weight distribution. The vehicle or compound was given 20 minutes before injection of substance P or saline, and the animals were returned to an acrylic cage consisting of 4 chambers (about 30x30x30cm, produced ad libitum) immediately after transdermal injection, allowed to acclimate and observed for a pruritic response 1 hour before the present experiment. A camera (Samsung, VLUUNV30) was positioned in the upper part of the mice to record their response. Pruritus was determined by blind counting of scratching times over a 20min period immediately following intradermal injection. The number of scratching was defined as 3 or more separate rapid scratching movements around the injection site using hind paws (J.Toxicol. Sci.2009, 34(4), 427-. All data were analyzed by using Excel and Prism, and all values were expressed as mean ± s.e.m. The inhibitory effect of the compound is shown as a percentage of the maximal response to histamine (control 100%). Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant. The inhibitory effect of the compounds is shown in table 10 below.
[ Table 10]
| Compound (I) | Number of scratching (% change from control group) |
| Compound 4 | 50.8 |
| Compound 7 | 60.51 |
| Compound 18 | 42.6 |
| Compound 24 | 74.51 |
| Compound 48 | 33.5 |
| Compound 95 | 80.71 |
"1" refers to statistical significance.
Experimental example 10: compound 48/80-induced pruritus model in ICR mice
[ animals ]
Female ICR mice (8 weeks old) were purchased from OrientBio co., Ltd. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment: causes of itching and measurement thereof
24 hours prior to this experiment, hair was cut using a clipper (8000AD, THRIVE) in the back mouth under isoflurane anesthesia. Compound 48/80(100 μ g) was dissolved in saline and a volume of 40 μ L was injected intradermally into the posterior mouth portion. Mice were given either vehicle (20% cyclodextrin in double distilled water) or compound by oral administration. The compound (50mg/kg) was dissolved in vehicle (20% cyclodextrin in double distilled water) and administered orally in a volume of 10 mL/kg.
Mice (n-8-10 mice/group) were divided into 3 groups (normal group, control group, and experimental group). Animals were randomly grouped according to similar weight distribution. The vehicle or compound was given 20 minutes before injection of compound 48/80 or saline, and the animals were returned to an acrylic cage consisting of 4 chambers (approximately 30x30x30cm, produced ad libitum) immediately after transdermal injection, allowed to acclimate and observed for a pruritic response 1 hour prior to this experiment. A camera (Samsung, VLUUNV30) was positioned in the upper part of the mice to record their response. Pruritus was determined by blind counting of scratching times over a 20min period immediately following intradermal injection. The number of scratching was defined as 3 or more separate rapid scratching movements around the injection site using hind paws (J.Pharmacol. Sci.2006, 100, 285-. All data were analyzed by using Excel and Prism, and all values were expressed as mean ± s.e.m. The inhibitory effect of the compound is shown as a percentage of the maximal response to histamine (control 100%). Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant. The inhibitory effect of the compounds is shown in table 11 below.
[ Table 11]
| Compound (I) | Number of scratching (% change from control group) |
| Compound 4 | 49.61 |
| Compound 18 | 50.91 |
| Compound 24 | 47.91 |
| Compound 48 | 30.3 |
| Compound 95 | 93.61 |
"1" refers to statistical significance.
Experimental example 11: oxazolone-induced model of atopic dermatitis in Balb/c mice
[ animals ]
Female Balb/c mice (6 weeks old, 20. + -.3 g) were purchased from OrientBio Co., Ltd. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment ]
On the day of the start (day 1), mice were sensitized by topical application of 50 μ L of 1% oxazolone in acetone and olive oil (4:1 ratio) solution on abdominal skin. On days 8, 10, 12, 15, 17, 19, 22, 24 and 26, 25 μ L of 0.2% oxazolone was administered topically to the inside of the right ear of each mouse. On days 8-28, the thickness of the right ear of the mice was measured by orally administering the compound of the present invention (10mL/kg) 2 times a day, in the morning and afternoon on days 12, 19 and 27. On day 28, mouse ear tissues were extracted, fixed in 10% formalin solution, and then tissue sections were prepared, stained with H & E (hematoxylin and eosin), and then observed for ear thickness (Journal of Investigative Dermatology, 2008, 128, 79-86). Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant. The inhibitory effect of the compounds is shown in table 12 below.
[ Table 12]
"1" refers to statistical significance.
Experimental example 12: determination of pharmacological effects in atopic dermatitis NC/Nga mice induced by in vitro administration of dermatophagoides farinae
[ animals ]
Female NC/Nga mice (3 weeks old, 15-17g) were purchased from Central Lab. Animals were housed under controlled temperature (23 + -3 deg.C), humidity (50 + -5%) and light and free access to food and water.
[ experiment ]
The mice were shaved with scissors on their backs and the remaining hair was completely removed with a shaving cream. To disrupt the barrier, 150 μ L of 4% Sodium Dodecyl Sulfate (SDS) was applied to the shaved back skin. The Df ointment (100 mg/mouse) was spread 2 times per week for 3 weeks. Compounds were administered orally in the morning and afternoon for 3 weeks, and then the severity of dermatitis was assessed on days 2,7, 10, 14, and 21. (1) Erythema/hemorrhage, (2) scarring/drying, (3) edema, (4) scratch/erosion were scored 0 (none), 1 (mild), 2 (moderate) and 3 (severe). The total skin score was defined as the sum of the individual scores (Scandinavian Journal of immunological 2011, 73, 536-. Statistical analysis was analyzed using one-way ANOVA and dunnett's test. Values with p <0.05 were considered statistically significant. The improvement effect of skin lesions is shown in table 13 below.
[ Table 13]
| Compound (I) | Skin damage score (day 21) |
| Normal group (non-Df) | 0.0 |
| Control group (Df) | 9.0 |
| Compound 24(25mg/kg, BID) | 7.2 |
| Compound 24(50mg/kg, BID) | 4.41 |
"1" refers to statistical significance.
As described above, the compounds of the present invention have significant improvements in solubility and metabolic stability compared to conventional drugs. As a result of comparative pharmacokinetic analysis using ICR mouse and SD rat animal models in comparison with the compound disclosed in WO2010/030785, the compound of the present invention showed excellent effects in terms of pharmacokinetic properties, such as AUC and maximum blood concentration which were much more than 7-8 times higher than the comparative compound. It was found that histamine-induced migration of inflammatory cells such as mast cells and eosinophils is mediated by The histamine 4 receptor (The Journal of Pharmacology and Experimental therapeutics, 2003, 305, 1212-1221), and that histamine 4 receptor antagonists may have an anti-inflammatory effect by inhibiting The increase of inflammatory cells. According to the improvement of pharmacokinetic properties, the compound of the present invention showed excellent pharmacological effects much more than 3 times higher than the compound disclosed in WO2010/030785, as shown by experimental results of inhibiting migration of histamine into mast cell trachea. Compared to the compounds disclosed in WO2010/030785, the compounds of the present invention showed strong anti-itch effects and the like in the histamine-induced itch model, and strong anti-inflammatory effects in the oxazolone-induced atopic model, which is one of the human body mimics (j.invest.dermatol.2008, 128, 79-86).
Furthermore, the compounds of the present invention show the same or stronger effect on the improvement of skin appearance in NC/Nga mouse atopic model, which is a human mimetic, compared to tacrolimus currently used as an atopic drug (j.clin.invest.1999, 104, 1097-1105). Therefore, it is expected to be extremely effective in treating atopic patients.
Claims (22)
1. Heterocyclic compounds of formula 1
[ formula 1]
Or a racemate, R-or S-isomer or a pharmaceutically acceptable salt thereof:
wherein
X1And X2Is C;
X3and X4Each independently isC or N, provided that X3And X4Is N;
R1is a saturated or unsaturated 3-12 membered mono-or poly-heterocyclyl containing 1-3 heteroatoms selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7And R8(ii) a Or R1is-NR6R7；
R2is-H; -C1-C6Alkyl or-halogen selected from-F, -Cl, -Br and-I;
R3is-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8An alkynyl group; -an amino group; -OH; -nitro (-NO)2)；-C6-C10An aryl group; or-heterocyclyl; wherein heterocyclyl is a saturated or unsaturated 3-6 membered heterocyclyl containing 1-3 heteroatoms selected from N, O and S;
R4is-H; -C1-C6An alkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; or-OH;
R5is-H;
with the proviso that when X3When is N, R4Is absent; and when X4When is N, R5Is absent;
Y1and Y5Each independently is C or N; and Y is2、Y3And Y4Each independently is C or a heteroatom independently selected from N, O and S, with the proviso that Y1、Y2And Y5Is N; or Y2、Y3And Y4At least two of which are heteroatoms independently selected from N, O and S;
Y2and Y3Each of which may be independently substituted by R9Substitution;
Y4can be substituted by-H or-C1-C6Alkyl substitution;
R6and R7Each independently selected from-H and-C1-C6An alkyl group;
R8is-C1-C6Alkyl or-C3-C8A cycloalkyl group; and is
R9Is selected from-H; -C1-C6Alkyl and-C3-C7A cycloalkyl group;
wherein the alkyl group may be unsubstituted or substituted by one or more substituents selected from-C1-C4Alkoxy and-OH.
2. The heterocyclic compound of claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein X3Is C or N, and X4Is N.
3. The heterocyclic compound of claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein Y1And Y5Each independently is C or N; and Y is2、Y3And Y4Each independently is C or a heteroatom independently selected from N, O and S, with the proviso that Y1、Y2、Y3、Y4And Y5Is N.
4. The heterocyclic compound of claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein R1Is a saturated or unsaturated 3-to 8-membered mono-or poly-heterocyclyl group containing 1 to 3 heteroatoms selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7And R8。
5. The heterocyclic compound of claim 1 or a pharmaceutically acceptable salt thereofA spiro, R-or S-isomer or a pharmaceutically acceptable salt thereof, wherein R is3Is selected from-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8Alkynyl and-OH.
6. The heterocyclic compound according to claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein
X1And X2Is a compound of formula (I) and (II),
X3and X4Each independently is C or N, provided that X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-12 membered mono-or poly-heterocyclyl containing 1-3 heteroatoms selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7And R8(ii) a Or R1is-NR6R7，
R2is-H; -C1-C6Alkyl or-halogen selected from-F, -Cl, -Br and-I;
R3is-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -C1-C6A perhaloalkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy; -C2-C7An alkenyl group; -C2-C8An alkynyl group; or an-OH group, or a group,
R4is-H; -C1-C6An alkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C1-C6Perhaloalkoxy radical(ii) a or-OH;
R5is-H;
with the proviso that when X3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1and Y5Each independently is C or N; and Y is2、Y3And Y4Each independently is C or a heteroatom independently selected from N, O and S, with the proviso that Y1、Y2And Y5Is N; or Y2、Y3And Y4At least two of which are heteroatoms independently selected from N, O and S,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
Y4can be substituted by-H or-C1-C6The substitution of the alkyl group is carried out,
R6and R7Each independently selected from-H and-C1-C6An alkyl group, a carboxyl group,
R8is selected from-C1-C6Alkyl and-C3-C8Cycloalkyl radicals, and
R9is selected from-H; -C1-C6Alkyl and-C3-C7A cycloalkyl group,
wherein alkyl may be unsubstituted or substituted with one or more substituents selected from-OH and-C1-C4An alkoxy group.
7. The heterocyclic compound according to claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein
X1And X2Is a compound of formula (I) and (II),
X3and X4Each independently is C or N, provided that X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-12 membered mono-or poly-heterocyclyl containing 1-3 heteroatoms selected from N, O and S, wherein R is1Unsubstituted or substituted by 1 to 3 substituents selected from-NR6R7And R8，
R2is-H; -C1-C6Alkyl or-halogen selected from-F, -Cl, -Br and-I;
R3is-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C2-C7An alkenyl group; or-C2-C8An alkynyl group,
R4is-H; -C1-C6An alkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; or-C1-C6A halogenated alkoxy group,
R5is a group of formula (I) having the formula-H,
with the proviso that when X3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1and Y5Each independently is C or N; and Y is2、Y3And Y4Each independently selected from C, N and O, with the proviso that Y1、Y2And Y5Is N; or Y2、Y3And Y4At least two of which are N or O,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6An alkyl group, a carboxyl group,
R8is selected from-C1-C6Alkyl radical, and
R9is selected from-H, -C1-C6Alkyl and-C3-C7A cycloalkyl group,
wherein alkyl may be unsubstituted or substituted with one or more substituents selected from-OH and-C1-C4An alkoxy group.
8. The heterocyclic compound according to claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein
X1And X2Is a compound of formula (I) and (II),
X3and X4Each independently is C or N, provided that X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-6 membered heterocyclic group containing 1-3 heteroatoms selected from N, O and S, wherein R1Unsubstituted or substituted by-NR6R7The substitution is carried out by the following steps,
R2is-H; -C1-C6Alkyl or-halogen selected from-F, -Cl, -Br and-I;
R3is-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; -C1-C6A haloalkoxy group; -C2-C7An alkenyl group; or-C2-C8An alkynyl group,
R4is-H; -C1-C6An alkyl group; -halogen selected from-F, -Cl, -Br and-I; -CN; -C1-C6An alkoxy group; or-C1-C6A halogenated alkoxy group,
R5is a group of formula (I) having the formula-H,
with the proviso that when X3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1and Y5Each independently is C or N; and Y is2、Y3And Y4Each independently selected from C, N and O, with the proviso that Y1、Y2And Y5At least two of which are N,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6Alkyl radical, and
R9is selected from-H, -C1-C6Alkyl and-C3-C7A cycloalkyl group,
wherein the alkyl group may be unsubstituted or substituted by one or more substituents selected fromfrom-OH and-C1-C4An alkoxy group.
9. The heterocyclic compound according to claim 1, or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein
X1And X2Is a compound of formula (I) and (II),
X3and X4Each independently is C or N, provided that X3And X4Is N is the number of N,
R1is a saturated or unsaturated 3-6 membered heterocyclic group containing 1-3 heteroatoms selected from N, O and S, wherein R1Unsubstituted or substituted by-NR6R7The substitution is carried out by the following steps,
R2is-H; -C1-C6Alkyl or-halogen selected from-F, -Cl, -Br and-I;
R3is-H; -C1-C6An alkyl group; -C1-C6A haloalkyl group; or-halogen selected from-F, -Cl, -Br and-I,
R4is-H; -C1-C6An alkyl group; or-halogen selected from-F, -Cl, -Br and-I,
R5is a group of formula (I) having the formula-H,
with the proviso that when X3When is N, R4Is absent; and when X4When is N, R5In the absence of the presence of the agent,
Y1and Y5Each independently is C or N; and Y is2、Y3And Y4Each independently selected from C, N and O, with the proviso that Y1、Y2And Y5At least two of which are N,
Y2and Y3Each of which may be independently substituted by R9The substitution is carried out by the following steps,
R6and R7Each independently selected from-H and-C1-C6Alkyl radical, and
R9is selected from-H and-C1-C6An alkyl group, a carboxyl group,
wherein alkyl may be unsubstituted or substituted with one or more substituents selected from-OH and-C1-C4An alkoxy group.
10. A heterocyclic compound selected from the group consisting of:
3-methyl-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 1);
8-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 2);
4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 3);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 4);
3-chloro-6- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 5);
6- (4-Methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazine (Compound 6);
8-chloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 7);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 8);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylpyrrolidin-3-amine (compound 9);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine (compound 10);
(R) -1- (3-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylpyrrolidin-3-amine (compound 11);
8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 12);
4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 13);
8-chloro-1-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 14);
8-chloro-1-methyl-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 15);
8-bromo-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 16);
7, 8-dichloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 17);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 18);
(S) -8-chloro-4- (3-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 19);
(S) -8-chloro-4- (3, 4-dimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 20);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile (compound 21);
8-chloro-4- (3,4, 5-trimethylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 22);
8-chloro-7-ethoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 23);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 24);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 25);
4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-8-carbonitrile (compound 26);
1- (7, 8-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 27);
8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine-7-carbonitrile (compound 28);
8-chloro-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 29);
(R) -1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) pyrrolidin-3-amine (compound 30);
9-chloro-2-methyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 31);
9-chloro-2-methyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 32);
1- (9-chloro-2-methylpyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 33);
9-chloro-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 35);
1- (9-chloropyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 36);
9-chloro-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 37);
9-chloro-2-cyclopropyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 38);
9-chloro-2-cyclopropyl-5- (piperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 39);
9-chloro-2- (methoxymethyl) -5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 40);
9-chloro-2-ethyl-5- (4-methylpiperazin-1-yl) pyrido [3,2-e ] [1,2,4] triazolo [1,5-c ] pyrimidine (compound 41);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine (compound 43);
1- (8-chloro-2-methyl-2H-pyrazolo [3,4-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine (compound 44);
8-chloro-2-methyl-4- (piperazin-1-yl) -2H-pyrazolo [3,4-c ] [1,8] naphthyridine (compound 45);
8-chloro-4- (4-methylpiperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine (compound 46);
8-chloro-4- (piperazin-1-yl) imidazo [1,2-a ] pyrido [2,3-e ] pyrazine (compound 47);
1- (8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 48);
8-iodo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 49);
n-methyl-1- (8-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 50);
1- (8- (difluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 51);
n-methyl-1- (8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 52);
4- (4-methylpiperazin-1-yl) -8- (trifluoromethyl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 53);
1- (8-ethynylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 54);
n-methyl-1- (8-vinylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 55);
1- (8-ethylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 56);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-ol (compound 57);
1- (8-methoxypyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 58);
1- (8- (difluoromethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 59);
8-chloro-7-methoxy-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 60);
8-chloro-7-methoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 61);
7, 8-dichloro-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 62);
8-chloro-7-ethoxy-4- (piperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 63);
8-chloro-4- (4-methylpiperazin-1-yl) -7- (2,2, 2-trifluoroethoxy) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 64);
1- (8-bromo-9-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 65);
8-bromo-9-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 66);
1- (8, 9-dichloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 67);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride (compound 68);
1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrochloride (compound 69);
8-chloro-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 70);
8-bromo-4- (5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 71);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -3-methylazetidin-3-amine (compound 72);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, 3-dimethylazetidin-3-amine (compound 73);
8-bromo-4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 74);
4- (hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl) -8-iodopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 75);
8-chloro-4- (4-cyclopropylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 76);
4- ((1S,4S) -2, 5-diazabicyclo [2.2.1] hept-2-yl) -8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 77);
8-chloro-4- ((1S,4S) -5-methyl-2, 5-diazabicyclo [2.2.1] hept-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 78);
8-chloro-4- (1, 4-diazepan-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 79);
(R) -1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylpyrrolidin-3-amine (compound 81);
8-chloro-4- (hexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (Compound 82);
8-chloro-4- (1-methylhexahydro-1H-pyrrolo [3,4-b ] pyridin-6 (2H) -yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 83);
1- (8-chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (compound 84);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N, N-dimethylazetidin-3-amine (compound 85);
2- ((8-Chloropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) amino) ethanol (Compound 87);
1- (8-chloroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 88);
1- (8-bromoimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 89);
(tert-butyl 1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) azetidin-3-yl) (methyl) carbamate (compound 90);
1- (8-chloro-2-methyloxazolo [4,5-c ] [1,8] naphthyridin-4-yl) -N-methylazetidin-3-amine (compound 91);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) oxazolo [4,5-c ] [1,8] naphthyridine (compound 92);
1- (8-chloropyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 93);
8-chloro-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine (compound 94);
1- (8-bromopyrido [2,3-e ] tetrazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 95);
8-bromo-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] tetrazolo [1,5-a ] pyrazine (compound 96);
1- (8-chloro-2-methylpyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 97);
8-chloro-2-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [1,5-a ] pyrazine (compound 98);
1- (8-bromo-7-methylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 99);
8-bromo-7-methyl-4- (4-methylpiperazin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazine (compound 100);
8-chloro-4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-7-ol HCl salt (compound 101);
n- (1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-yl) -N-methylhydroxylamine (compound 102);
1- (8-Bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2,2, 2-trifluoroacetate (Compound 104);
(S) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 105);
(R) -1- (8-bromo-2-methyl-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 106);
1- (8-bromo-1, 2-dihydroimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 107);
1- (8-bromo-2-methylimidazo [1,2-a ] pyrido [2,3-e ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 108);
1- (9-bromo-2-methylpyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 109);
1- (9-bromopyrazolo [1,5-c ] pyrido [3,2-e ] pyrimidin-5-yl) -N-methylazetidin-3-amine (compound 110);
n-methyl-1- (8-nitropyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 111);
4- (3- (methylamino) azetidin-1-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-8-amine (compound 112);
n-methyl-1- (8-phenylpyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) azetidin-3-amine (compound 113);
1- (8- (furan-2-yl) pyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 114);
1- (2-bromopyrido [2,3-e ] pyrrolo [1,2-a ] pyrazin-6-yl) -N-methylazetidin-3-amine (compound 115);
1- (2-chloropyrido [3,2-e ] [1,2,4] triazolo [4,3-a ] pyrazin-6-yl) -N-methylazetidin-3-amine (compound 116);
1- (8-chloropyrido [3,4-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (compound 117);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine methanesulfonate (compound 118);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine maleate (compound 119);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 2-hydroxypropane-1, 2, 3-tricarboxylate (compound 120);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine nitrate (compound 121);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydroiodide (compound 122);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine phosphate (compound 123);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine 4,4' -methylenebis (3-hydroxy-2-naphthoic acid) salt (compound 124);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine hydrobromide (compound 125);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine sulfate (compound 126);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (2R,3R) -2, 3-dihydroxysuccinate salt (compound 127);
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (1S) - (+) -10-camphorsulfonate (compound 128); and
1- (8-bromopyrido [2,3-e ] [1,2,4] triazolo [4,3-a ] pyrazin-4-yl) -N-methylazetidin-3-amine (S) -2-hydroxypropionate (compound 130);
or a racemate, an R-or S-isomer or a pharmaceutically acceptable salt thereof.
11. A process for preparing a heterocyclic compound having the structure of formula 1 as defined in claim 1 or its racemate, R-or S-isomer or pharmaceutically acceptable salt, which comprises the steps of:
(a) by using the formula R1Arylating a compound of formula 4 with a compound of formula H to prepare a compound of formula 3;
(b) preparing a compound of formula 2 by arylating the prepared compound of formula 3; and
(c) cyclizing the prepared compound of formula 2:
[ formula 2]
[ formula 3]
[ formula 4]
Wherein in the formulae 2 to 4, X1、X2、X3、X4、R1、R2、R3、R4、R5、R9And Y5The same as defined in formula 1; and Z represents a reactive leaving group.
12. A process for preparing a heterocyclic compound having the structure of formula 5 or its racemate, R-or S-isomer or pharmaceutically acceptable salt, wherein said formula 5 is wherein Y1、Y2And Y4Formula 1 as defined in claim 1, which is N, the method comprising the steps of:
(a) preparing a compound of formula 9 by halogenating a compound of formula 10;
(b) preparing a compound of formula 8 by cyanating the prepared compound of formula 9;
(c) preparing a compound of formula 7 by acylating the prepared compound of formula 8;
(d) preparing a compound of formula 6 by cyclizing the prepared compound of formula 7 followed by halogenation; and
(e) using the formula R1-H arylating the prepared compound of formula 6:
[ formula 5]
[ formula 6]
[ formula 7]
[ formula 8]
[ formula 9]
[ formula 10]
Wherein in formulae 5-10, X1、X2、X3、X4、R1、R2、R3、R4、R5、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group.
13. A process for preparing a heterocyclic compound having the structure of formula 11 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein formula 11 is wherein Y2、Y4And Y5Formula 1 as defined in claim 1, which is N, the method comprising the steps of:
(a) preparing a compound of formula 16 by arylating a compound of formula 17;
(b) preparing a compound of formula 15 by imidizing the prepared compound of formula 16;
(c) preparing a compound of formula 14 by acylating the prepared compound of formula 15;
(d) preparing a compound of formula 13 by cyclizing the prepared compound of formula 14;
(e) preparing a compound of formula 12 by reducing the prepared compound of formula 13, followed by halogenation; and
(f) using the formula R1Arylating the prepared compound of formula 12 with a compound of-H:
[ formula 11]
[ formula 12]
[ formula 13]
[ formula 14]
[ formula 15]
[ formula 16]
[ formula 17]
Wherein in formulae 11-17, X1、X2、X3、X4、R1、R2、R3、R4、R5、R8And R9The same as defined in formula 1; and Z represents a reactive leaving group.
14. A process for the preparation of a heterocyclic compound having the structure of formula 18 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein formula 18 is wherein Y2And Y3Formula 1 as defined in claim 1, which is N, the method comprising the steps of:
(a) preparing a compound of formula 21 by friedel-crafts reaction of a compound of formula 23 with a compound of formula 22;
(b) preparing a compound of formula 20 by cyclizing the prepared compound of formula 21;
(c) preparing a compound of formula 19 by halogenating the prepared compound of formula 20; and
(d) using the formula R1Arylating the prepared compound of formula 19 with a compound of-H:
[ formula 18]
[ formula 19]
[ formula 20]
[ formula 21]
[ formula 22]
[ formula 23]
Wherein in the formulae 18 to 23, X1、X2、X3、X4、R1、R2、R3、R4、R5And R9The same as defined in formula 1; and Z represents a reactive leaving group.
15. A process for the preparation of a heterocyclic compound having the structure of formula 24 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein formula 24 is wherein Y1、Y3And Y4Formula 1 as defined in claim 1, which is N, the method comprising the steps of:
(a) preparing a compound of formula 28 by cyclizing a compound of formula 29;
(b) preparing a compound of formula 27 by halogenating the prepared compound of formula 28 followed by hydroxylation;
(c) by using the formula R1Arylating the prepared compound of formula 27 with a compound of-H, followed by halogenation to prepare a compound of formula 26;
(d) preparing a compound of formula 25 by arylating the prepared compound of formula 26; and
(e) cyclizing the prepared compound of formula 25:
[ formula 24]
[ formula 25]
[ formula 26]
[ formula 27]
[ formula 28]
[ formula 29]
Wherein in the formulae 24 to 29, X1、X2、X3、X4、R1、R2、R3、R4、R5And R9The same as defined in formula 1; and Z represents a reactive leaving group.
16. A process for preparing a heterocyclic compound having the structure of formula 30 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein formula 30 is wherein Y2Is N and Y4Formula 1 as defined in claim 1, which is O, the process comprising the steps of:
(a) preparing a compound of formula 36 by esterifying a compound of formula 37;
(b) preparing a compound of formula 35 by arylating the prepared compound of formula 36;
(c) preparing a compound of formula 34 by cyclizing the prepared compound of formula 35;
(d) preparing a compound of formula 33 by enolization addition reaction of the prepared compound of formula 34;
(e) preparing a compound of formula 32 by cyclizing the prepared compound of formula 33;
(f) preparing a compound of formula 31 by halogenating the prepared compound of formula 32; and
(g) using the formula R1Arylating the prepared compound of formula 31 with a compound of-H:
[ formula 30]
[ formula 31]
[ formula 32]
[ formula 33]
[ formula 34]
[ formula 35]
[ formula 36]
[ formula 37]
Wherein in the formulae 30 to 37, X1、X2、X3、X4、R1、R2、R3、R4、R5、R6And R9The same as defined in formula 1; p represents a protecting group; and Z represents a reactive leaving group.
17. A process for the preparation of a heterocyclic compound having the structure of formula 38 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, said formula 38 being wherein Y1Is N and Y2And Y3Formula 1 as defined in claim 1, each independently being C or N, the process comprising the steps of:
(a) preparing a compound of formula 40 by suzuki coupling reaction of a compound of formula 42 with a compound of formula 41;
(b) preparing a compound of formula 39 by halogenating the prepared compound of formula 40; and
(c) using the formula R1Arylating the prepared compound of formula 39 with a compound of-H:
[ formula 38]
[ formula 39]
[ formula 40]
[ formula 41]
[ formula 42]
Wherein in the formulae 38 to 42, X1、X2、X3、X4、R1、R2、R3、R4And R5The same as defined in formula 1; and Z represents a reactive leaving group.
18. A process for the preparation of a heterocyclic compound having the structure of formula 43 or a racemate, R-or S-isomer or pharmaceutically acceptable salt thereof, wherein formula 43 is wherein Y5Formula 1 as defined in claim 1, which is N, the method comprising the steps of:
(a) preparing a compound of formula 46 by pyrrolation of a compound of formula 47;
(b) preparing a compound of formula 45 by cyclizing the prepared compound of formula 46;
(c) preparing a compound of formula 44 by halogenating the prepared compound of formula 45; and
(d) using the formula R1Arylating the prepared compound of formula 44 with a compound of-H:
[ formula 43]
[ formula 44]
[ formula 45]
[ formula 46]
[ formula 47]
Wherein in the formulae 43 to 47, X1、X2、X3、X4、R1、R2、R3、R4And R5The same as defined in formula 1; and Z represents a reactive leaving group.
19. A pharmaceutical composition comprising an effective amount of a compound as defined in any one of claims 1 to 10 or a racemate, R-or S-isomer or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.
20. The pharmaceutical composition of claim 19, wherein the composition exhibits human histamine 4 receptor (hH4R) inhibitory activity.
21. The pharmaceutical composition of claim 19, wherein the composition is for treating or preventing a disease selected from the group consisting of inflammatory diseases, autoimmune diseases, allergic diseases, eye diseases, skin diseases, respiratory diseases, pain diseases, heart diseases, and human histamine 4 receptor (hH4R) -related diseases.
22. The pharmaceutical composition of claim 21, wherein the composition is used for treating or preventing a disease selected from the group consisting of nasal polyps, allergic rhinitis, non-allergic rhinitis, viral rhinitis, nasal pruritus, sinusitis, nasal congestion, asthma, Chronic Obstructive Pulmonary Disease (COPD), rheumatoid arthritis, atopic dermatitis, psoriasis, eczema, pruritus, cutaneous pruritus, urticaria, idiopathic chronic urticaria, scleroderma, conjunctivitis, keratoconjunctivitis, ocular inflammation, dry eye disease, cardiac dysfunction, arrhythmia, atherosclerosis, multiple sclerosis, inflammatory bowel disease, inflammatory pain, neuropathic pain, osteoarthritic pain, autoimmune thyroid disease, immune-mediated diabetes, lupus, postoperative adhesions, cancer, and vestibular disorders.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2011-0100369 | 2011-09-30 | ||
| KR10-2012-0033444 | 2012-03-30 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1196820A HK1196820A (en) | 2014-12-24 |
| HK1196820B true HK1196820B (en) | 2018-04-27 |
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