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HK1216532B - Novel compounds for selective histone deacetylase inhibitors, and pharmaceutical composition comprising the same - Google Patents

Novel compounds for selective histone deacetylase inhibitors, and pharmaceutical composition comprising the same Download PDF

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Publication number
HK1216532B
HK1216532B HK16104539.2A HK16104539A HK1216532B HK 1216532 B HK1216532 B HK 1216532B HK 16104539 A HK16104539 A HK 16104539A HK 1216532 B HK1216532 B HK 1216532B
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formula
methyl
group
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HK16104539.2A
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HK1216532A1 (en
Inventor
李昌植
梁贤模
崔好辰
金度勳
金昭映
河妮娜
林孝珍
高恩熙
尹成爱
斐大权
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株式会社钟根堂
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Priority claimed from PCT/KR2014/003776 external-priority patent/WO2014178606A1/en
Publication of HK1216532A1 publication Critical patent/HK1216532A1/en
Publication of HK1216532B publication Critical patent/HK1216532B/en

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Description

Novel compounds for selective histone deacetylase inhibitors and pharmaceutical compositions comprising the same
Technical Field
The present invention relates to novel compounds having Histone Deacetylase (HDAC) inhibitory activity, optical isomers thereof, pharmaceutically acceptable salts thereof, their use for the preparation of medicaments for the treatment of HDAC mediated diseases, pharmaceutical compositions comprising the same, methods of treatment using the compositions, and methods for the preparation thereof.
Background
Cellular transcriptional regulation is a complex biological process. One of the basic principles is the post-translational modification of the histone proteins H2A/B, H3 and H4 to form an octameric histone core complex. Complex N-terminal modifications at lysine residues by acetylation or methylation and at serine residues by phosphorylation constitute part of the so-called "histone code" (Stahl & Ellis, Nature 403, 41-45, 2000).
In a simple model, acetylation of positively charged lysine residues reduces the affinity for negatively charged DNA, so transcription factors can easily enter.
Histone acetylation and deacetylation are catalyzed by Histone Acetyltransferase (HAT) and Histone Deacetylase (HDAC), respectively. HDACs are associated with transcription repression complexes that switch chromatin to transcriptionally inactive, silent structures (Marks et al Nature Cancer Rev 1, 189-202, 2001). For certain HATs associated with the transcriptional activation complex, and vice versa. Three different classes of HDACs located in the nucleus have been described so far, namely class I (HDAC 1-3, 8; Mr. gtoreq.42-55 kDa) sensitive to the inhibition of Trichostatin a (TSA), class II (HDAC 4-7, 9, 10; Mr. gtoreq.120-130 kDa) sensitive to TSA, and class III (Sir2) which are very different by their NAD + dependency and TSA insensitivity.
Inhibitors of Histone Deacetylase (HDAC) constitute a new class of anticancer agents with differentiation and apoptotic activity. By targeting Histone Deacetylases (HDACs), HDAC inhibitors affect histone (protein) acetylation and chromatin structure, inducing complex transcriptional reprogramming, such as reactivation of tumor suppressor genes and suppression of oncogenes. In addition to acetylation of the N-terminal lysine residues produced in the core histone protein, there are non-histone targets important for cancer cell biology, including heat shock protein (HSP90), tubulin or p53 tumor suppressor protein. Therefore, HDAC inhibitors may be used for the treatment of cancer as well as inherited metabolic diseases, autoimmune diseases, etc., since efficacy in animal models has been shown for inflammatory diseases, rheumatoid arthritis and neurodegeneration.
Examples of histone deacetylase-mediated diseases include cell proliferative diseases including malignant tumors such as cancer and the like, autosomal dominant diseases such as Huntington's disease and the like, hereditary metabolic diseases such as cystic fibrosis (cystic fibrosis), hepatic fibrosis, renal fibrosis (kidney fibrosis), pulmonary fibrosis (pulmonary fibrosis), dermal fibrosis and the like, autoimmune diseases (autoimmune diseases) such as rheumatoid arthritis and the like, acute and chronic neurological diseases (acute and chronic neurological diseases) such as diabetes (diabetes), stroke (stroke) and the like, hypertrophy (cardiac hypertrophy) such as cardiac hypertrophy and the like, hemorrhagic heart failure (hemorrhagic heart failure), amyotrophic lateral sclerosis (amyotrophic lateral sclerosis), glaucoma (glaucomia), ocular disease (ocular disease) (associated with angiogenesis), Alzheimer's disease, and the like.
The HDAC inhibitors known to date can be divided into four classes according to their structure: 1) short chain fatty acids (butyric acid, valproic acid); 2) hydroxamic acid (trichostatin a, SAHA, LBH-589); 3) cyclic peptides (desipeptide); and 4) benzamide (MS-275, MGCD-0103) (International Journal of Onocology 33, 637-646, 2008). These many HDAC inhibitors (SAHA, LBH-589, MS-275, etc.) effectively induce growth inhibition, differentiation and apoptosis of various transformed cells in culture as well as in animal models (Marks, p.a, etc., Curr Opin oncol.2001.13.477-483), and some HDAC inhibitors such as SAHA, LBH-589, MS-275, etc., were clinically evaluated for the treatment of various cancers (Johnstone, R.W nat. rev. drug discov.20021.287-299). Typical examples of HDAC inhibitor compounds known to date include hydroxamate compounds such as SAHA (U.S. Pat. No. 771,760, Zolinza, Vorinostat), PXD101(WO 02/30879, Belinostat), LBH-589(WO 02/22577, Panobinostat) and benzamide compounds such as MS-275(EP8799) and MGCD0103(WO 04/69823). Among these compounds, SAHA was approved in 2006, month 10 and has been used for the treatment of CTCL (cutaneous T-cell lymphoma). The diseases for which drugs are effective have additionally expanded, but defects in efficacy and side effects are known (Cancer Res2006, 66, 5781-5789).
That is, although many HDAC inhibitors have been reported so far, most HDAC inhibitors have defects in terms of efficacy and side effects. Therefore, in order to overcome these drawbacks, there is a continuing need to develop effective HDAC inhibitors with high selectivity and low side effects (Mol Cancer Res, 5, 981, 2007).
Disclosure of Invention
Technical problem
An object of the present invention is to provide a novel compound, an optical isomer thereof or a pharmaceutical salt thereof.
It is another object of the present invention to provide a pharmaceutical composition comprising a novel compound having a highly selective Histone Deacetylase (HDAC) inhibitory activity, an optical isomer thereof, or a pharmaceutical salt thereof.
It is still another object of the present invention to provide a method for preparing the same.
It is still another object of the present invention to provide a pharmaceutical composition comprising the above compound for the treatment of diseases associated with HDAC activity, including cancer, inflammatory diseases, autoimmune diseases, neurological or neurodegenerative diseases, and the like.
It is a further object of the present invention to provide the use thereof for the preparation of a medicament for the treatment of HDAC mediated diseases such as cancer, inflammatory diseases, autoimmune diseases, neurological or neurodegenerative diseases and the like.
It is still another object of the present invention to provide a method for treating HDAC mediated diseases such as cancer, inflammatory diseases, autoimmune diseases, neurological or neurodegenerative diseases, etc., which comprises administering a therapeutically effective amount of a pharmaceutical composition comprising the above compound.
Means for solving the problems
The inventors of the present invention have found a novel compound having Histone Deacetylase (HDAC) inhibitory activity and used it for the treatment of histone deacetylase mediated diseases, thereby completing the present invention.
In accordance with the above objects, the present invention provides a compound of the following formula I:
[ formula I ]
Wherein A is
Xa and Xb are each independently C or N,
L1and L2Each independently of the others is hydrogen, -F, -Cl, -Br, -I, -CF3or-C1-3A linear or branched alkyl group,
q is C (═ O), S ((═ O)2) S (═ O), or C (═ NH),
y is selected from the group consisting of:
m is C, O, N, S (═ O)2Or a combination of the above, or S,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4A linear or branched alkyl group (wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I); a saturated or unsaturated 5-to 7-membered heterocyclic compound comprising as ring members from 1 to 3 heteroatoms selected from N, O and S (wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I); phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group;
wherein R iscAnd RdEach independently of the other being hydrogen, C1-3A linear or branched alkyl group;or
(wherein as the alkyl group substituted with the heterocyclic compound or the heterocyclic compound mentioned in Ra1And Ra2The heterocyclic compound in (1) is preferably pyridine, morpholine or tetrahydropyran
n is an integer of 0, 1 or 2,
Rbis hydrogen; a hydroxyl group; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CF3(ii) a -F; -Cl; -Br; -I; orWherein R iseAnd RfEach independently is hydrogen or-C1-3A linear or branched alkyl group, and
z is phenyl; or a saturated or unsaturated 5-to 7-membered heterocyclic ring containing as ring members from N, O and 1 to 3 heteroatoms of S, wherein the phenyl or heterocyclic ring of Z may be unsubstituted or substituted with at least one hydroxyl group; -C1-4A linear or branched alkyl group which is unsubstituted or has one or more substituents; -CF3Unsubstituted or substituted with one or more substituents; -CN, which is unsubstituted or has one or more substituents; -C1-6A linear or branched alkoxy group which is unsubstituted or has one or more substituents; -C2-6A straight-chain or branched alkylalkoxy group which is unsubstituted or has one or more substituents; -C1-3An alcohol which is unsubstituted or has one or more substituents; phenyl which is unsubstitutedSubstituted or substituted with one or more substituents; a heterocycle, which is unsubstituted or has one or more substituents; or halogen, which is unsubstituted or has one or more substituents.
Preferably, Z may be selected from the group consisting of:
wherein P isaAnd PbEach independently isHydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F; or-C1-3The alcohol is added into the mixture of the alcohol,
whereinSelected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan, tetrahydropyridine, piperidine or the following groups:
x, y and z are each independently an integer of 0 or 1, and
Rg1、Rg2and Rg3Each independently selected from hydrogen; a hydroxyl group; -C1-3An alkyl group; -CF3；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -N (CH)3)2(ii) a -F; -Cl; -Br; -I; benzene and its derivativesA group; -S ((═ O)2)CH3(ii) a Or the following groups:
preferably, the compound represented by the above formula I may be a compound represented by the following formula II:
[ formula II ]
Wherein
a is an integer of 0, 1 or 2,
k is independently hydrogen, -F, -Cl, -Br or-I,
y is selected from the group consisting of:
m is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4Straight or branched chain alkyl (wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3，CH3，CH2CH3F, Cl, Br or I); saturated or unsaturated 5-to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from N, O and S as ring members(wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3，CH3，CH2CH3F, Cl, Br or I); phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group;wherein R iscAnd RdEach independently of the other being hydrogen, C1-3A linear or branched alkyl group;or
n is an integer of 0, 1 or 2,
Rbis hydrogen; a hydroxyl group; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CF3(ii) a -F; -Cl; -Br; -I; orWherein R iseAnd RfEach independently is hydrogen or-C1-3A linear or branched alkyl group, and
z is selected from the group consisting of:
wherein P isaAnd PbEach independently isHydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F; or-C1-3The alcohol is added into the mixture of the alcohol,
whereinSelected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan, tetrahydropyridine, piperidine or the following groups:
the indazole of (a) can be indazol-5-yl or indazol-6-yl,
the pyridine of (a) may be a pyridin-3-yl group,
the indole of (a) may be indol-4-yl, indol-5-yl or indol-6-yl,
the pyrimidine in (b) may be a pyrimidin-5-yl group,
the quinoline of (a) may be quinolin-7-yl,
the furan of (a) may be a furan-3-yl group,
the tetrahydropyridine of (a) may be 1, 2, 3, 6-tetrahydropyridin-4-yl,
the piperidine of (a) may be a piperidin-4-yl group,
is/are as followsMay be benzo [ d ]][1，3]A dioxol-5-yl group,
is/are as followsMay be a 3, 6-dihydro-2H-pyran-4-yl group,
is/are as followsMay be tetrahydro-2H-pyran-4-yl,
is/are as followsMay be 2, 3-dihydrobenzo [ b ]][1，4]IIAn in-6-yl group, or a pharmaceutically acceptable salt thereof,
x, y and z are each independently an integer of 0 or 1, and
Rg1、Rg2and Rg3Each independently selected from hydrogen; a hydroxyl group; -C1-3An alkyl group; -CF3；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -N (CH)3)2(ii) a -F; -Cl; -Br; -I; a phenyl group; -S ((═ O)2)CH3(ii) a Or the following groups:
in an embodiment according to the present invention, the compound represented by formula II above may preferably include the following structure:
wherein
a is an integer of 0, 1 or 2,
k is independently hydrogen, -F, -Cl, -Br or-I,
y is
M is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4A linear or branched alkyl group (wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I); a saturated or unsaturated 5-to 7-membered heterocyclic compound comprising as ring members from 1 to 3 heteroatoms selected from N, O and S (wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I); phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group; or
n is an integer of 0, 1 or 2,
Rbis hydrogen; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; and is
Z is selected from the group consisting of:
wherein P isaAnd PbEach independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F；-C1-An alcohol;or
May be benzo [ d ]][1，3]A dioxol-5-yl group,
may be 2, 3-dihydrobenzo [ b ]][1，4]IIIn-6-yl.
More preferably, the compound represented by the above formula II may be a compound that may be represented by the following formula III:
[ formula III ]
Wherein
Y is
M is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; by phenyl, pyridine or pyrimidine substituted-C1-4A linear or branched alkyl group (wherein the phenyl, pyridine or pyrimidine may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I);
n is an integer of 0, 1 or 2,
Rbis hydrogen; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; and is
Z is selected from the group consisting of:
wherein P isaAnd PbEach independently is hydrogen; -F; -Cl; -Br; -CF3；-OCF3；-CH2F；Or
May be benzo [ d ]][1，3]A dioxol-5-yl group,
may be 2, 3-dihydrobenzo [ b ]][1，4]IIIn-6-yl.
The compound represented by the above formula I may be a compound represented by the following formula I-1:
[ formula I-1]
Wherein R is1To R5Each independently of the others is hydrogen, -F, -Cl, -Br, -I, -CF3，-CH3，-CH2CH3，-OCH3or-OCF3And is and
R6is piperidine; by one or more C1-3An alkyl-substituted piperidine; morpholine; by one or more C1-3Alkyl substituted morpholines; piperazine; by one or more C1-3Alkyl-substituted piperazines, -C (═ O) CH3Benzhydryl, phenyl, benzyl, pyridine, -C1-2An alkyl morpholine, or a morpholinoethanone (wherein the phenyl or benzyl group may be unsubstituted or at least one hydrogen may optionally be-OCH3，-CH3，-CH2CH3-F, -Cl, -Br, or-I substitution); (piperazinyl) ethanone; (R) -3-fluoropyrrolidine; (S) -3-fluoropyrrolidine; (R) -pyrrolidin-2-yl-methanol; (S) -2- (trifluoromethyl) pyrrolidine; azetidine; difluoroazetidine; phenyl piperidinol; or oxazepane (oxazepane).
Preferably, the compound represented by the above formula I-1 may be a compound 255, 256, 279, 374, 385, 386, 389, 390, 391, 392, 393, 394, 413, 414, 415, 416, 438, 439, 440, 441, 453, 454, 455, 456, 457, 458, 459, 460, 461, 477, 478, 479, 480, 481, 482, 484, 485, 486, 494, 495, 496, 497, 498, 520, 521, 522, 529, 530, 543, 544, 545, 580, 683, 684, 717, 718, 771, 772, 773, 774, 776, 791, 800 or 801 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-2:
[ formula I-2]
Wherein R is4Is indole, pyridine, difluorophenyl, (dimethylamino) pyridine, pyrimidine, bis (trifluorophenyl) phenyl, 2, 3-dihydrobenzo [ b ]][1，4]IIEnglish, trimethoxyphenyl, dimethylphenyl, furan, 3, 6-dihydro-2H-pyran, 1-methyl-1, 2, 3, 6-tetrahydropyridine, 1-Boc-1, 2, 3, 6-tetrahydropyridine, benzo [ d][1，3]Dioxoles, or tetrahydro-2H-pyrans.
Preferably, the compound represented by the above formula I-2 may be compound 261, 262, 263, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 418 or 483 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-3:
[ formula I-3]
Wherein R is4Is 1-C1-3alkyl-1H-indazole and R6Is morpholine, piperidine or piperazine, wherein said piperazine is unsubstituted or substituted by-C1-3Alkyl substitution.
Preferably, the compound represented by the above formula I-3 may be a compound 252, 253, 254 or 260 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-4:
[ formula I-4]
Wherein R is4Is tetrahydropyridine or piperidine and R7Is absent (nothing), acetyl, methylsulfonyl, N-isopropylcarbamoyl, 2-hydroxy-2-methylpropyl, 2-fluoro-2-methylpropyl, tert-butyl carboxylate.
Preferably, the compound represented by the above formula I-4 may be a compound 419, 420, 489, 490, 491, 492, 493, 517 or 518 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-5:
[ formula I-5]
Wherein R is6Is piperidine, piperidinol, dimethylmorpholine, phenylpiperidinol, methylpiperazine, 2- (piperazin-1-yl) ethanol, pyridine, (S) -pyrrolidin-2-ylmethanol, cyclopropyl (piperazin-1-yl) methanol, or azetidine.
Preferably, the compound represented by the above formula I-5 may be a compound 462, 463, 464, 465, 466, 467, 468, 469, 470 or 471 as described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-6:
[ formula I-6]
Wherein R is3And R4Each independently of the others is hydrogen, -F, -Cl, -Br, -I, -CF3Benzo [ d ]][1，3]Dioxole or dihydro-2H-pyran, and R6Is morpholine, hydroxypiperidine or difluoroazetidine.
Preferably, the compound represented by formula I-6 above may be compound 487, 488, 511, 512, 513, 514, 532, 577, or 578 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-7:
[ formula I-7]
Wherein P, Q1And Q2Each independently is C or N, R3And R4Each independently of the others is hydrogen, -F, -Cl, -Br, -I, or-CF3And R is6Is piperidine, substituted by one or more C1-3Piperidine, morpholine, substituted by one or more C1-3Morpholine substituted by alkyl, phenyltetrahydropyridine, by one or more C1-3Alkyl-substituted piperazines, benzylpiperazines, phenylpiperidinols, (methoxyphenyl) piperazines, (fluorophenyl) piperazines, pyrrolidines, substituted with one or more C1-3Alkyl-substituted diazepans, azetidines, (dimethylphenyl) piperazines, (1, 4-diazepan-1-yl) ethanones, cyclopropyl (piperazin-1-yl) methanones, or fluoropyrrolidines.
Preferably, the compound represented by the above formula I-7 may be a compound 280, 281, 309, 311, 312, 313, 341, 342, 343, 352, 353, 354, 355, 356, 357, 358, 376, 377, 379, 450, 451, 533, 778, 826, 827, 828, or 829 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-8:
[ formula I-8]
Wherein R is3Is 1-methyl-1H-indazole, phenyl, difluorophenyl, pyridine, pyrimidine, quinoline, biphenyl, indole, trimethoxyphenyl, bis (trifluoromethyl) phenyl, -F, -Cl, -Br, or-I.
Preferably, the compound represented by the above formula I-8 may be a compound 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340 or 372 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-9:
[ formula I-9]
Wherein R is7Is morpholine, substituted by one or more C1-3Alkyl substituted piperazine, (S) -pyrrolidin-2-ylmethanol, or (methylamino) ethanol.
Preferably, the compound represented by the above formula I-9 may be compound 380, 381, 382 or 383 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-10:
[ formula I-10]
Wherein R is8Is hydroxy, -F, -Cl, -Br, or-I and R6Is morpholine, ethylpiperazine, or piperazinyl-2-fluoro-2-methylpropyl.
Preferably, the compound represented by the above formula I-10 may be compound 499, 500, 765 or 766 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-11:
[ formula I-11]
Wherein R is9Is hydrogen, hydroxy, -F, -Cl, -Br, or-I.
Preferably, the compound represented by the above formula I-11 may be the compound 370 or 371 described in the present invention.
The compound represented by the above formula I may be a compound represented by the following formula I-12:
[ formula I-12]
Wherein R is6May be selected from the following groups:
preferably, the compound represented by the above formula I-12 may be compound 531, 651, 716, 797, 802 or 803 described in the present invention.
The compounds represented by formula I and I-1 to I12 are as follows:
in the present invention, pharmaceutically acceptable salts refer to salts commonly used in the pharmaceutical industry, such as inorganic ion salts with calcium, potassium, sodium, magnesium, etc., inorganic acid salts with hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, tartaric acid, sulfuric acid, etc., organic acid salts with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc., sulfonic acid salts with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc., amino acid salts with glycine, arginine, lysine, etc., and amine salts with trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the type of salt used in the present invention is not limited to the above-mentioned salts.
In the present invention, a preferred salt is a hydrochloride, and preferred examples of this compound include compounds 461 and 493 described in the present specification.
Specific preparation methods of the novel compounds of formula I, optical isomers thereof, or pharmaceutically acceptable salts thereof are shown in the following reaction schemes 1 to 12.
[ reaction scheme 1]
In reaction scheme 1, the compound of formula 1-2 is synthesized by reductive amination of the compound of formula 1-1 with methyl 4-formylbenzoate at room temperature for 12 to 24 hours or with methyl 4- (bromomethyl) benzoate at room temperature for 16 hours, and the compound of formula 1-3 is synthesized by reacting the compound of formula 1-2 with 4-nitrophenyl chloroformate at room temperature for 12 to 24 hours.
Then, the compounds of formulae 1-4 are prepared by reacting the compounds of formulae 1-3 with an amine derivative (R)6) In the presence of a dimethylformamide solvent at room temperature or at 60 ℃ for 1 to 2 days. Finally, the target compound 255, 256, 279, 374, 385, 386, 389, 390, 391, 392, 393, 394, 413, 414, 415, 416, 438, 439, 440, 441, 453, 454, 455, 456, 457, 458, 459, 460, 461, 477, 478, 479, 480, 481, 482, 484, 485, 486, 494, 495, 496, 497, 498, 520, 521, 522, 529, 530, 543, 544, 545, 580, 683, 684, 717, 718, 771, 773, 774, 776, 791, 800 or 801 by reacting a compound of formula 1-4 with potassium hydroxide (KOH), methanol and hydroxylamine hydrochloride (HONH)2HCl) at room temperature or with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 2]
In reaction scheme 2, a compound of formula 2-2 is synthesized by a Suzuki reaction of a compound of formula 2-1 with boronic acid, and then a compound of formula 2-4 is synthesized by reduction of a compound of formula 2-3 with hydrogen and palladium. Finally, the target compound 261, 262, 263, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 418 or 483 is prepared by dissolving the compound of formula 2-2 or 2-4 in methanol, followed by reaction with hydroxylamine hydrochloride (HONH)2HCl) and potassium hydroxide (KOH) at room temperature, or with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 3]
In reaction scheme 3, a compound of formula 3-2 is synthesized by a Suzuki reaction of a compound of formula 3-1 with boronic acid, and then a compound of formula 3-3 is synthesized by reacting a compound of formula 3-2 with an amine derivative (R)6) Reacting at 50-60 deg.c. Finally, the target compound 252, 253, 254 or 260 was synthesized by dissolving the compound of formula 3-3 in methanol, followed by reaction with hydroxylamine hydrochloride, potassium hydroxide (KOH) and an aqueous solution of hydroxylamine (50 wt%) (which were added dropwise) at room temperature.
[ reaction scheme 4]
In reaction scheme 4, the compound of formula 4-2 is synthesized by reacting the compound of formula 4-1 with hydrogen chloride, and then the compound of formula 4-3 is synthesized using acetic anhydride or methanesulfonyl chloride. Next, the compound of formula 4-4 was synthesized by reducing the compound of formula 4-1 with hydrogen and palladium, and the compound of formula 4-6 was synthesized by the same method. Subsequently, the compound of formula 4-7 is synthesized by reacting the compound of formula 4-5 with 2, 2-dimethyloxirane at 120 ℃ in a microwave reactor, and then the compound of formula 4-8 is synthesized using diethylaminosulfur trifluoride (DAST). Finally, the target compound 419, 420, 489, 490, 491, 492, 493, 517 or 518 is prepared by dissolving a compound of formula 4-3, 4-6, 4-4, 4-7 or 4-8 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 5]
In reaction scheme 5, the compound of formula 5-2 is synthesized by Suzuki reaction of the compound of formula 5-1 with boric acid, and then the compound of formula 5-3 is synthesized by reacting formula 5-2 with 4-nitrophenyl chloroformate at room temperature. Next, the compound of formula 5-4 is prepared by reacting the compound of formula 5-3 with an amine derivative (R)6) Reacted at 50 ℃ to synthesize the product. Finally, target compound 462, 463, 464, 465, 466, 467, 468, 469, 470 or 471 is prepared by dissolving a compound of formula 5-4 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 6]
In reaction scheme 6, the compound of formula 6-2 is synthesized by substituting the compound of formula 1-1 with the compound of formula 6-1, and then the compound of formula 6-3 is synthesized by hydrolyzing the compound of formula 6-2 with lithium hydroxide (LiOH). Next, the compound of formula 6-4 is prepared by reacting the compound of formula 6-3 with methanol, 1-ethyl-3- [ 3-dimethylaminopropyl group]Carbodiimide hydrochloride (EDC) and 1-hydroxybenzotriazole hydrate (HOBt), and then the compound of formula 6-5 is synthesized by reacting the compound of formula 6-4 with 4-nitrophenyl chloroformate at room temperature. Subsequently, the compound of formula 6-6 is prepared by reacting the compound of formula 6-5 with an amine derivative (R)6) Synthesized by reaction at 50 deg.c, and then, if necessary, a compound of formula 6-7 is synthesized by Suzuki reaction of a compound of formula 6-6 with boric acid. Finally, the target compound 487, 488, 511, 512, 513, 514, 532, 577 or 578 is prepared by dissolving the compound of formula 6-6 or 6-7 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 7]
In reaction scheme 7, the compound of formula 7-4 is synthesized by reductive amination of the compound of formula 7-3 or by substitution of the compound of formula 7-1 with the compound of formula 7-2. Then, the compound of formula 7-5 is synthesized by reacting the compound of formula 7-4 with 4-nitrophenyl chloroformate at room temperature or at 60 ℃. Subsequently, the compound of formula 7-6 is prepared by reacting the compound of formula 7-5 with an amine derivative (R)6) At 60 ℃ or by adding acetyl chloride to the compound of formula 7-4. Finally, the target compound 280, 281, 309, 311, 312, 313, 341, 342, 343, 352, 353, 354, 355, 356, 357, 358, 376, 377, 379, 450, 451, 533, 778, 826, 827, 828 or 829 is produced byDissolving the compound of formula 7-6 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 8]
In reaction scheme 8, a compound of formula 8-2 is synthesized by substituting a compound of formula 8-1 with aniline, and then a compound of formula 8-3 is synthesized by reacting the compound of formula 8-2 with N-bromosuccinimide at room temperature. Next, the compound of formula 8-4 was synthesized by reacting the compound of formula 8-3 with 4-nitrophenyl chloroformate at room temperature, and then the compound of formula 8-5 was synthesized by substituting the compound of formula 8-4 with morpholine at 60 ℃. Subsequently, the compound of formula 8-6 was synthesized by Suzuki reaction of the compound of formula 8-5 with boric acid. Finally, the target compound 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340 or 372 is prepared by dissolving a compound of formula 8-5 or 8-6 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 9]
In reaction scheme 9, a compound of formula 9-1 is synthesized by a Suzuki reaction of a compound of formula 8-5 with 4-formylphenylboronic acid, and then a compound of formula 9-2 is synthesized by reacting the compound of formula 9-1 with an amine derivative (R)7) Reductive amination and synthesis. Finally, the target compound 380, 381, 382 or 383 was prepared by dissolving the compound of formula 9-2 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 10]
In reaction scheme 10, the compound of formula 10-2 is synthesized by reductive amination of the compound of formula 10-1 with methyl 4-formylbenzoate, and the compound of formula 10-3 is synthesized by reacting the compound of formula 10-2 with 4-nitrophenyl chloroformate at room temperature. Next, the compound of formula 10-4 was synthesized by substituting the compound of formula 10-3 with morpholine at room temperature, and then the compound of formula 10-5 was synthesized by hydrolyzing the compound of formula 10-4 with lithium hydroxide (LiOH). Subsequently, the compound of formula 10-6 is synthesized by reacting the compound of formula 10-5 with (tetrahydropyran-2-yl) hydroxylamine, 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) and 1-hydroxybenzotriazole hydrate (HOBt) at room temperature. Then, target compound 499 was synthesized by dissolving the compound of formula 10-6 in methanol and adding hydrogen chloride at room temperature.
The compound of formula 10-7 is synthesized by reacting the compound of formula 10-4 with diethylaminosulfur trifluoride (DAST), and then the target compound 500 is synthesized by dissolving the compound of formula 10-7 in methanol, followed by hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
The compound of formula 10-8 is synthesized by substituting the compound of formula 10-3 with ethylpiperazine, and then the compound of formula 10-9 is synthesized using diethylaminosulfur trifluoride (DAST). Next, the objective compound 765 was prepared by dissolving the compound of formula 10-9 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
The compound of formula 10-11 was synthesized by introducing Boc-piperazine to the compound of formula 10-3, followed by reaction with diethylaminosulfur trifluoride (DAST). The compounds of formulas 10-12 were then synthesized by Boc-deprotection under acidic conditions. Next, compounds of formula 10-13 are prepared by reacting compounds of formula 10-12The compound is synthesized by reacting with 2, 2-dimethyloxirane under microwave irradiation, and then the compound of formula 10-14 is synthesized using diethylaminosulfur trifluoride (DAST). Finally, the target compound 766 is prepared by dissolving the compound of formula 10-14 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 11]
In reaction scheme 11, a compound of formula 11-3 is synthesized by substituting a compound of formula 11-2 with a compound of formula 11-1 twice, and then a compound of formula 11-5 is synthesized by cyclizing a compound of formula 11-3 with a compound of formula 11-4 in the presence of an ethanol solvent by stirring under reflux. Next, the compound of formula 11-6 was synthesized by reacting the compound of formula 11-5 with 4-nitrophenyl chloroformate at room temperature, and then the compound of formula 11-7 was synthesized by substituting the compound of formula 11-6 with morpholine at 60 ℃. Finally, the target compound 370 or 731 is prepared by dissolving the compound of formula 11-7 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
[ reaction scheme 12]
In reaction scheme 12, the compound of formula 12-1 is synthesized by reacting the compound of formula 1-3 with N-Boc-piperazine, and then the compound of formula 12-2 is synthesized by Boc-deprotection under acidic conditions. Next, the compound of formula 12-3 is synthesized by reacting the compound of formula 12-2 with 2, 2-dimethyloxirane under microwave irradiation, and then the compound of formula 12-4 is synthesized using diethylaminosulfur trifluoride (DAST). Then, Compound 531 is prepared by combining a compound of formula 12-4The solution was dissolved in methanol and then reacted with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
A compound of formula 12-5 is prepared by reacting a compound of formula 12-2 with 1, 6-dioxaspiro [2, 5]]Octane is synthesized by reaction under microwave irradiation, and then compound 651 is synthesized by dissolving the compound of formula 12-5 in methanol, followed by hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
The compound of formula 12-6 is synthesized by reacting the compound of formula 12-5 with DAST, and then compound 716 is synthesized by dissolving the compound of formula 12-6 in methanol, followed by hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
Compounds of formulae 12-8 are prepared by reacting an amine derivative (R)6) Introduced into the compound of formula 1-3, followed by substitution, and then the target compound 797, 802 or 803 is synthesized by dissolving the compound of formula 12-8 in methanol, followed by reaction with hydroxylamine (NH)2OH) aqueous solution and potassium hydroxide (KOH), which are successively added dropwise, at room temperature.
The present invention provides a pharmaceutical composition for preventing or treating histone deacetylase-mediated diseases, comprising a compound represented by the following formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
[ formula I ]
Wherein Y, Q, Z and A are as defined above.
Examples of histone deacetylase-mediated diseases include cell proliferative diseases including malignancies such as cancer and the like, inflammatory diseases, autosomal dominant diseases such as Huntington's disease and the like, hereditary metabolic diseases such as cystic fibrosis (cystic fibrosis), hepatic fibrosis, renal fibrosis (kidney fibrosis), pulmonary fibrosis (pulmonary fibrosis), dermal fibrosis (skin fibrosis) and the like, autoimmune diseases such as rheumatoid arthritis and the like, acute and chronic neurological diseases such as diabetes (diabetes), stroke (stroke) and the like, hypertrophy such as cardiac hypertrophy, hemorrhagic sclerosis such as cardiac hypertrophy, ischemic sclerosis (ischemic sclerosis) and the like, ischemic cardiovascular diseases such as glaucoma, ischemic cardiovascular diseases such as ischemic cardiovascular diseases, ischemic cardiovascular diseases such as the development of the eye, and the like, Neurodegenerative diseases (neurodegenerative diseases), and the like, and also includes symptoms and diseases associated with abnormal functions of histone deacetylases.
The compound represented by formula I of the present invention may be one of the compounds represented by the above formulas I-1 to I-12, formula II or formula III.
The pharmaceutically acceptable salts are the same as those described for the compounds represented by formula I above.
For administration, the pharmaceutical composition of the present invention may further comprise one or more pharmaceutically acceptable carriers in addition to the compound represented by formula I above, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. Examples of pharmaceutically acceptable carriers include saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and combinations thereof. Other conventional additives such as antioxidants, buffer solutions, antibacterial agents, etc. may be added to the composition if desired. In addition, the pharmaceutical composition of the present invention can be formulated into injections such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules or tablets by adding diluents, dispersants, surfactants, binders and lubricants. Thus, the compositions of the present invention may be presented as patches, liquids, pills, capsules, granules, tablets, suppositories, and the like. Depending on the disease and/or the composition, these preparations may be formulated by suitable methods in the art or as described in Remington's pharmaceutical science (latest edition), Mack Publishing Company, Easton PA.
The composition of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) in one dose depending on various factors including patient body weight, age, sex, health state, diet, administration time, administration route, excretion rate, disease severity, and the like. The daily dose of the composition of the present invention may be about 1 to 500mg/kg, preferably 5 to 100mg/kg, and the composition of the present invention may be administered once or several times per day.
In addition to the compound represented by the above formula I, its optical isomer, or a pharmaceutically acceptable salt thereof, the pharmaceutical composition of the present invention may further comprise one or more active ingredients having the same or similar potency.
The present invention provides a method for preventing or treating histone deacetylase-mediated diseases, which comprises administering a therapeutically effective amount of a compound represented by the above formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
As used herein, the term "therapeutically effective amount" refers to an amount of a compound represented by formula I above that is effective for preventing or treating histone deacetylase-mediated diseases.
Further, the present invention provides a method for inhibiting Histone Deacetylase (HDAC), which comprises administering an effective amount of a compound represented by the above formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof to mammals including humans.
The method for preventing or treating histone deacetylase-mediated diseases according to the present invention includes managing the diseases themselves and inhibiting or avoiding the symptoms by administering the compound represented by the above formula I before the onset of the symptoms. In the management of disease, the prophylactic or therapeutic dose of a particular active ingredient may vary depending on the nature and severity of the disease or condition and the route of administration of the active ingredient. The dose and dose frequency may vary depending on the age, weight and response of the individual patient. Based on these factors, appropriate dosage and use can be readily selected by one of ordinary skill in the art. In addition, the method for preventing or treating histone deacetylase-mediated diseases according to the present invention may further include administering a therapeutically effective amount of an additional active ingredient, which together with the compound represented by the above formula I, improves the treatment of diseases. The additional active ingredients may exhibit a synergistic or additive effect in conjunction with the compound represented by formula I above.
The present invention also provides the use of a compound represented by the above formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of HDAC mediated diseases. The compound represented by the above formula I for the preparation of a medicament may contain pharmaceutically acceptable adjuvants, diluents, carriers, etc., and may be formulated into a combined preparation together with other active ingredients to have synergistic activity.
The references to use, compositions and methods of treatment of the present invention are equally applicable unless they are mutually inconsistent.
Advantageous effects
The compound represented by the above formula I, its optical isomer or a pharmaceutically acceptable salt thereof according to the present invention may selectively inhibit HDAC, and thus have an excellent effect on the prevention and treatment of histone deacetylase-mediated diseases.
Description of the drawings
Figure 1 shows the acetylation levels of tubulin, histone H3 and histone H4 after treatment with compound 255.
Figure 2 shows the acetylation levels of tubulin, histone H3 and histone H4 after treatment with compound 374.
Figure 3 shows the improvement of arthritis in the collagen-induced arthritis model after treatment with compound 254 or compound 255.
Figure 4 shows the improvement of arthritis in the collagen-induced arthritis model after treatment with compound 374.
Figure 5 shows the improvement of arthritis in the adjuvant-induced arthritis model after treatment with compounds 255, 374 or 461.
Figure 6 shows the recovery of body weight in colitis model after treatment with compound 254 or 255.
Figure 7 shows the combination index after combined treatment of compound 255 and bortezomib (Velcade).
EMBODIMENTS FOR CARRYING OUT THE INVENTION
Hereinafter, the present invention will be described further specifically with reference to examples, preparation examples and test examples. It is to be understood, however, that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Unless otherwise indicated, the reagents and solvents mentioned below are those available from Sigma-Aldrich and TCI, HPLC is Waters e2695, silica gel for column chromatography is Merck (230-400 mesh),1h NMR data were measured using Bruker400MHz and the mass spectra were an Agilent 1100 series.
Preparation of novel urea compounds
The preparation of the compound of formula 1 is described by the reaction scheme.
Example 1: synthesis of Compound 252
(formula 1-2: 4- ((3-bromophenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (3-bromoaniline; 5g, 29mmol) was dissolved in dichloromethane (200mL), then methyl 4-formylbenzoate (5.7g, 35mmol) and acetic acid (1.74mL, 29mmol) were added and stirred for 1 hour. Then, sodium cyanoborohydride (2.2g, 35mmol) was slowly added dropwise and stirred for 1 day. Water was poured into the reaction mixture, and the organic layer was extracted. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (7.8g, 84%) as a white solid.
(formula 3-1: 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((3-bromophenylamino) methyl) benzoate; 5.90g, 18.4mmol) was dissolved in acetonitrile (200mL), followed by addition of potassium carbonate (5.09g, 36.9mmol) and 4-nitrophenyl chloroformate (4.09g, 20.3 mmol). Then, the mixture was heated and stirred at 50 ℃ for 1 day. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 1%) and concentrated to give the desired compound of formula 3-1 (7.3g, 82%) as a white solid.
(formula 3-2: 4- (((3- (1-methyl-1H-indazol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 3-1 (methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 1.00g, 2.06mmol), 1-methyl-1H-indazol-5-ylboronic acid (0.435g, 2.473mmol), Pd (dppf) Cl2(0.168g, 0.206mmol) and carbonic acidSodium (0.693g, 4.53mmol) and dimethoxyethane (5mL)/H2O (5mL) was mixed, heated at 120 ℃ for 15 minutes under microwave irradiation, and then the temperature was lowered to room temperature. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 3-2 (0.620g, 56%) as a white solid.
(formula 3-3: 4- ((N- (3- (1-methyl-1H-indazol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (1-methyl-1H-indazol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.05g, 0.093mmol) of the compound of formula 3-2 was dissolved in dimethylformamide (10mL), followed by addition of morpholine (0.012mL, 0.14mmol) and potassium carbonate (0.039g, 0.28 mmol). Then, the mixture was heated and stirred at 60 ℃ for 12 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 3-3 (0.035g, 77%) as a colorless liquid.
(Compound 252: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1-methyl-1H-indazol-5-yl) phenyl) morpholine-4-carboxamide)
A compound of formula 3-3 (methyl 4- ((N- (3- (1-methyl-1H-indazol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.035g, 0.072mmol) was dissolved in methanol (10mL), followed by dropwise addition of hydroxylamine hydrochloride (0.025g, 0.361mmol), potassium hydroxide (0.041g, 0.722mmol) and hydroxylamine (50% by weight in water; 0.186mL, 1.445mmol) and stirring at room temperature for 6 hours. After completion of the reaction, methanol was removed under pressure, and then 2N hydrochloric acid was added to precipitate a solid. Then, the resulting solid was filtered and dried to obtain the desired compound 252(0.016g, 46%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.08(s，1H)，7.97(s，1H)，7.69-7.64(m，4H)，7.43-7.35(m，5H)，7.08(d，1H，J＝7.0Hz)，4.93(s，2H)，4.05(s，3H)，3.40-3.37(m，4H)，3.19-3.16(m，4H)；MS(ESI)m/z 486.1(M++H)。
Example 2: synthesis of Compound 253
(formula 3-3: 4- ((4-methyl-N- (3- (1-methyl-1H-indazol-5-yl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (1-methyl-1H-indazol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.05g, 0.093mmol) of the compound of formula 3-2 was dissolved in dimethylformamide (10mL), followed by addition of n-methylpiperazine (0.016mL, 0.14mmol) and potassium carbonate (0.039g, 0.28 mmol). Then, the mixture was heated and stirred at 55 ℃ for 12 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formula 3-3 (0.044g, 95%) as a colorless liquid.
(Compound 253: N- (4- (hydroxycarbamoyl) benzyl) -4-methyl-N- (3- (1-methyl-1H-indazol-5-yl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4-methyl-N- (3- (1-methyl-1H-indazol-5-yl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.044g, 0.088mmol, of the compound of formula 3-3 was dissolved in methanol (5mL), followed by dropwise addition of hydroxylamine hydrochloride (0.031g, 0.442mmol), potassium hydroxide (0.049g, 0.884mmol) and hydroxylamine (50% by weight aqueous solution; 0.228mL, 1.77mmol) and stirring at room temperature for 6 hours. After completion of the reaction, methanol was removed under pressure, and then 2N hydrochloric acid was added to precipitate a white solid. Then, the resulting solid was filtered and dried to obtain the desired compound 253(0.02g, 45%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.08(s，1H)，7.97(s，1H)，7.69-7.64(m，4H)，7.41-7.37(m，5H)，7.07-7.06(m，1H)，4.92(s，2H)，4.06(s，3H)，3.20-3.19(m，4H)，2.13-2.11(m，4H)，2.06(s，3H)；MS(ESI)m/z 499.2(M++H)。
Example 3: synthesis of Compound 254
(formula 3-2: 4- (((3- (1-methyl-1H-indazol-6-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 3-1 (methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 1.00g, 2.06mmol), 1-methyl-1H-indazol-6-ylboronic acid (0.435g, 2.47mmol), Pd (dppf) Cl2(0.168g, 0.206mmol) and sodium carbonate (0.693g, 4.53mmol) with dimethoxyethane (5mL)/H2O (5mL), heated at 120 ℃ for 15 minutes under microwave irradiation,the temperature was then reduced to room temperature. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 3-2 (1.03g, 93%) as a white solid.
(formula 3-3: 4- ((N- (3- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (1-methyl-1H-indazol-6-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.05g, 0.093mmol) of the compound of formula 3-2 was dissolved in dimethylformamide (10mL), followed by addition of morpholine (0.011mL, 0.14mmol) and potassium carbonate (0.0386g, 0.28 mmol). Then, the mixture was heated and stirred at 60 ℃ for 12 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 3-3 (0.035g, 78%) as a white solid.
(Compound 254: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamide)
A compound of formula 3-3 (methyl 4- ((N- (3- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.035g, 0.072mmol) was dissolved in methanol (5mL) and then hydroxylamine hydrochloride (0.025g, 0.361mmol), potassium hydroxide (0.041g, 0.722mmol) and hydroxylamine (50% by weight in water; 0.186mL, 1.45mmol) were added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under pressure, and then 2N hydrochloric acid was added to precipitate a solid. The resulting solid was then filtered and dried to give the desired compound 254(0.024g, 68%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.04(s，1H)，7.83(s，1H)，7.80(d，1H，J＝8.4Hz)，7.65(d，2H，J＝8.2Hz)，7.52(s，1H)，7.48(d，1H，J＝7.9Hz)，7.43-7.38(m，4H)，7.14(d，1H，J＝7.9Hz)，4.95(s，2H)，4.09(s，2H)，3.41-3.40(m，4H)，3.19-3.18(m，4H)；MS(ESI)m/z 486.1(M++H)。
Example 4: synthesis of Compound 255
(formula 1-4: 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, formula 1-3, 1.5g, 3.09mmol was dissolved in acetonitrile (50mL) and potassium carbonate (1.28g, 9.3mmol) and morpholine (0.40mL, 4.64mmol) were slowly added. Then, the temperature was slowly raised and the mixture was stirred at 80 ℃ for 3 hours. The temperature was lowered to room temperature, and dimethylformamide (50ml) was further added and stirred at 80 ℃ for 5 hours. Then, the reaction was completed, and the organic layer was washed three times with a saturated aqueous ammonium chloride solution, dehydrated with sodium sulfate, and filtered. Then, the filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) to give the desired compound of formulae 1 to 4 (0.45g, 33.6%) as a transparent oil.
(Compound 255: N- (3-bromophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.05g, 0.12mmol, of the compound of formula 1-4 was dissolved in methanol (2ml), and hydroxylamine hydrochloride (0.040g, 0.58mmol) was slowly added. Then, potassium hydroxide (0.065g, 1.15mmol) was added and stirred at room temperature for 10 minutes, and hydroxylamine (50.0 wt% aqueous solution; 0.14mL, 2.31mmol) was added. The mixture was stirred at room temperature for 1 day, and then the organic solvent was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. Then, the organic layer was washed three times with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered, and then the filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0-80%) to give the desired compound 255(0.036g, 72%) as a white solid.
1H NMR(400MHz，CDCl3-d6)δ7.63(d，2H，J＝7.8Hz)，7.27-7.20(m，4H)，7.13(t，1H，J＝7.8Hz)，6.96(d，1H，J＝7.1Hz)，4.83(s，2H)，3.49(brs，4H)，3.23(brs，4H)；MS(ESI)m/z436(M++H)。
Example 5: synthesis of Compound 256
(formula 1-4: 4- ((N- (3-bromophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, formula 1-3, 1.5g, 3.09mmol, was dissolved in acetonitrile (50mL) and potassium carbonate (1.28g, 9.3mmol) and 1-methylpiperazine (0.52mL, 4.64mmol) were slowly added. Then, the temperature was slowly raised and the mixture was stirred at 80 ℃ for 3 hours. The temperature was lowered to room temperature, and dimethylformamide (50ml) was further added and stirred at 80 ℃ for 5 hours. Then, the reaction was completed, and the organic layer was washed three times with a saturated aqueous ammonium chloride solution, dehydrated with sodium sulfate, and filtered. Then, the filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) to give the desired compound of formulae 1 to 4 (0.9g, 65%) as a transparent oil.
(Compound 256: N- (3-bromophenyl) -N- (4- (hydroxycarbamoyl) benzyl) methylpiperazine-1-carboxamide)
Methyl 4- ((N- (3-bromophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoate, a compound of formula 1-4 (0.05g, 0.12mmol) was dissolved in methanol (2ml), and hydroxylamine hydrochloride (0.039g, 0.56mmol) was slowly added. Then, potassium hydroxide (0.063g, 1.12mmol) was added and stirred at room temperature for 10 minutes, and hydroxylamine (50.0 wt% aqueous solution; 0.14mL, 2.24mmol) was added. The mixture was stirred at room temperature for 1 day, and then the organic solvent was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. Then, the organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered to obtain the desired compound 256(0.05g, 99%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.2Hz)，7.39(d，2H，J＝8.0Hz)，7.30-7.09(m，4H)，4.91(brs，2H)，3.26(m，4H)，2.26(m，4H)，2.22(s，3H)；MS(ESI)m/z 449(M++H)。
Example 6: synthesis of Compound 260
(formula 3-3: 4- ((N- (3- (1-methyl-1H-indazol-5-yl) phenyl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (1-methyl-1H-indazol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.05g, 0.093mmol) of the compound of formula 3-2 was dissolved in dimethylformamide (3mL), followed by addition of piperidine (0.012mL, 0.14mmol) and potassium carbonate (0.039g, 0.28 mmol). Then, the mixture was heated and stirred at 60 ℃ for 12 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 30%) and concentrated to give the desired compound of formula 3-3 (0.042g, 93.4%) as a colorless liquid.
(Compound 260: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1-methyl-1H-indazol-5-yl)
Phenyl) piperidine-1-carboxamide
Methyl 4- ((N- (3- (1-methyl-1H-indazol-5-yl) phenyl) piperidine-1-carboxamido) methyl) benzoate, a compound of formula 3-3 (0.042g, 0.087mmol) was dissolved in methanol (5mL), then hydroxylamine hydrochloride (0.030g, 0.435mmol), potassium hydroxide (0.049g, 0.87mmol) were added, and hydroxylamine (50 wt% aqueous solution; 0.224mL, 1.74mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under pressure, and then 2N hydrogen chloride was added to precipitate a solid. The resulting solid was then filtered and dried to give the desired compound 260(0.036g, 85.5%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.08(s，1H)，7.95(s，1H)，7.71-7.64(m，4H)，7.37-7.36(m，5H)，7.04-7.03(m，1H)，4.90(s，2H)，4.05(s，3H)，3.18-3.17(m，4H)，1.42-1.41(m，2H)，1.29-1.28(m，4H)；MS(ESI)m/z 483.56(M++H)。
Example 7: synthesis of Compound 261
(formula 2-2: 4- ((N- (3- (1H-indol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.12g, 0.28mmol) was placed in a microwave reactor, followed by dimethoxyethane (5 mL). Then, indole-6-boronic acid (0.054g, 0.33mmol) and Pd (dppf) Cl were slowly added2(0.023g, 0.028mmol) followed by addition of saturated aqueous sodium carbonate solution (0.093g, 0.61 mmol). The mixture was stirred at 120 ℃ for 15 minutes under microwave irradiation, and then the reaction was completed. After washing three times with saturated aqueous sodium chloride solution, the organic layer was dehydrated with sodium sulfate, filtered, and concentrated. Then, the filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 80%) and concentrated to give the desired compound of formula 2-2 (0.091g, 70%) as a white solid.
(Compound 261: N- (3- (1H-indol-6-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1H-indol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.09g, 0.19mmol) was dissolved in methanol (10mL) and hydroxylamine hydrochloride (0.067g, 0.96mmol) was slowly added. Then, potassium hydroxide (0.11g, 1.92mmol) was added and stirred at room temperature for 10 minutes, and then hydroxylamine (50.0 wt% aqueous solution; 0.23mL, 3.83mmol) was added and stirred at room temperature for 1 day. The organic solvent was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. The resulting solid was then filtered and dried to give the desired compound 261(0.05g, 55%) as a yellow solid.
1H NMR(400MHz，CDCl3-d6)δ7.65(d，2H，J＝8.2Hz)，7.60-7.57(m，2H)，7.40-7.36(m，6H，J＝7.8Hz)，7.24(d，1H，J＝8.2Hz)，7.09-7.07(m，1H)，6.43(s，1H)，4.93(s，2H)，3.50(m，4H)，4.93(m，4H)，2.49(s，3H)；MS(ESI)m/z 471(M++H)。
Example 8: synthesis of Compound 262
(formula 2-2: 4- ((N- (3- (pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.11g, 0.25mmol) was placed in a microwave reactor, followed by dimethoxyethane (5 mL). Then, pyridine-3-boronic acid (0.037g, 0.31mmol) and Pd (dppf) Cl were slowly added2(0.021g, 0.025mmol) followed by addition of saturated aqueous sodium carbonate solution (0.085g, 0.56 mmol). The mixture was stirred at 120 ℃ for 15 minutes under microwave irradiation, and then the reaction was completed. After washing three times with saturated aqueous sodium chloride solution, the organic layer was dehydrated with sodium sulfate, filtered, and concentrated. Then, the filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 80%) and concentrated to give the desired compound of formula 2-2 (0.064g, 58%) as a white solid.
(Compound 262: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (pyridin-3-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.06g, 0.14mmol, of the compound of formula 2-2 was dissolved in methanol (10mL), followed by slow addition of hydroxylamine hydrochloride (0.048g, 0.70 mmol). Then, potassium hydroxide (0.078g, 1.40mmol) was added and stirred at room temperature for 10 minutes, and then hydroxylamine (50.0 wt% aqueous solution; 0.17mL, 2.78mmol) was added and stirred at room temperature for 1 day. The organic solvent was concentrated under reduced pressure and then neutralized by adding 2N hydrochloric acid. The organic layer was washed with diethyl ether and three times with saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered to obtain the desired compound 262(0.045g, 75%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.86(s，1H)，8.64(d，1H，J＝4.0Hz)，8.29(d，1H，J＝8.1Hz)，7.24-7.26(m，9H)，5.02(m，4H)，3.30(m，4H)；MS(ESI)m/z 433(M++H)。
Example 9: synthesis of Compound 263
(formula 2-2: 4- ((N- (3- (1H-indol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.05g, 0.12mmol) was placed in a microwave reactor, followed by dimethoxyethane (5 mL). Then, indole-5-boronic acid (0.023g, 0.14mmol) and Pd (dppf) Cl were slowly added2(0.009g, 0.012mmol) followed by addition of saturated aqueous sodium carbonate solution (0.039g, 0.25 mmol). Subjecting the mixture to microwaveThe reaction was completed after stirring at 120 ℃ for 15 minutes under irradiation. After washing three times with saturated aqueous sodium chloride solution, the organic layer was dehydrated with sodium sulfate, filtered, and concentrated. Then, the filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 80%) and concentrated to give the desired compound of formula 2-2 (0.040g, 74%) as a white solid.
(Compound 263N- (3- (1H-indol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1H-indol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.040g, 0.085mmol) was dissolved in methanol (2mL) and hydroxylamine hydrochloride (0.030g, 0.43mmol) was added slowly. Then, potassium hydroxide (0.48g, 0.85mmol) was added and stirred at room temperature for 10 minutes, and then hydroxylamine (50.0 wt% aqueous solution; 0.11mL, 1.70mmol) was added and stirred at room temperature for 1 day. The organic solvent was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. Then, the resulting solid was filtered and dried to obtain the desired compound 263(0.03g, 82%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.99(brs，1H)，7.75(s，1H)，7.64(d，1H，J＝7.8Hz)，7.44-7.05(m，9H)，6.46(s，1H)，4.93(s，2H)，3.39(m，4H)，3.17(m，4H)；MS(ESI)m/z 471(M++H)。
Example 10: synthetic Compound 279
(formula 1-2: 4- ((phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (aniline; 0.20g, 2.15mmol) was dissolved in dichloroethane (10mL), and triethylamine (0.45mL, 3.22mmol) was slowly added. Methyl 4- (bromomethyl) benzoate (0.74g, 3.22mmol) was then added, the temperature was slowly raised to 60 ℃ and the mixture was stirred for 1 hour. The reaction was completed, and the organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) to give the desired compound of formula 1-2 (0.15g, 29%) as a yellow oil.
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((phenylamino) methyl) benzoate; 0.15g, 0.62mmol) was dissolved in acetonitrile (3mL), followed by the slow addition of potassium carbonate (0.17g, 1.24mmol) and stirring at room temperature for 10 minutes. 4-Nitrophenyl chloroformate (0.14g, 0.68mmol) was then added, the temperature was slowly raised to 50 ℃ and the mixture was stirred for 12 hours. After completion of the reaction, the solvent was evaporated under reduced pressure, and the ethyl acetate layer was washed three times with a saturated aqueous sodium chloride solution, and then the organic layer was dehydrated with sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0-20%) to give the desired compound of formulae 1-3 (0.18g, 71%) as a transparent oil.
(formula 1-3: 4- ((N- (phenylmorpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (phenyl) amino) methyl) benzoate; 0.18g, 0.44mmol) of the compound of formulae 1-4 was dissolved in dimethylformamide (5mL), followed by the slow addition of potassium carbonate (0.18g, 1.33mmol) and morpholine (0.060g, 0.67 mmol). The temperature was lowered to room temperature, and dimethylformamide (50ml) was further added. Then, the temperature was slowly raised to 80 ℃, and the mixture was stirred for 3 hours. Then, the temperature was raised to 80 ℃ again, and the mixture was stirred for 5 hours. The reaction was completed, and the organic layer was washed three times with saturated aqueous ammonium chloride solution, dehydrated with sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) to give the desired compound of formulae 1 to 4 (0.10g, 64%) as a pale yellow solid.
(Compound 279: N- (4- (hydroxycarbamoyl) benzyl) -N-phenylmorpholine-4-carboxamide)
The compound of formula 1-4 (methyl 4- ((N-phenylmorpholine-4-carboxamido) methyl) benzoate; 0.15g, 0.42mmol) was dissolved in methanol (5ml) and hydroxylamine hydrochloride (0.15g, 2.12mmol) was added slowly. Then, potassium hydroxide (0.24g, 4.23mmol) was added and stirred at room temperature for about 10 minutes, and then hydroxylamine (50.0 wt% aqueous solution; 0.56mL, 8.46mmol) was added and stirred at room temperature for 1 day. The organic solvent was concentrated under reduced pressure, washed twice with diethyl ether after addition of water, neutralized by addition of 2N hydrochloric acid, and then further extracted with ethyl acetate. Then, the organic layer was washed twice with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered to obtain the desired compound 279(0.080g, 53%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ7.66(d，2H，J＝8.1Hz)，7.41(d，2H，J＝8.1Hz)，7.34(t，2H，J＝7.8Hz)，7.15(d，2H，J＝8.4Hz)，4.92(s，2H)，3.46(t，4H，J＝4.7Hz)，3.22(t，4H，J＝4.7Hz)；MS(ESI)m/z 356(M++H)。
Example 11: synthesis of Compound 280
(formula 7-4: 4- ((pyridin-2-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 7-3 (pyridin-2-amine, 0.2g, 2.13mmol) was dissolved in methanol (10mL), and then methyl 4-formylbenzoate (0.35g, 2.13mmol) was added and stirred at room temperature for 20 minutes. Then, sodium cyanoborohydride (0.13g, 2.13mmol) and acetic acid (0.12mL, 2.13mmol) were added slowly and stirred at room temperature for 5 hours. The organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0-30%) to give the desired compound of formula 7-4 (0.10g, 19%) as a transparent oil.
1H NMR(400MHz，CDCl3)δ8.17(d，1H，J＝5.8Hz)，8.06(d，2H，J＝8.4Hz)，7.66(t，1H，J＝7.8Hz)，7.44(d，2H，J＝8.0Hz)，6.76(t，1H，J＝6.7Hz)，6.58(d，1H，J＝8.6Hz)，4.67(d，2H，J＝6.0Hz)，3.92(s，3H)
(formula 7-5: 4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoic acid methyl ester)
Methyl (4- ((pyridin-2-ylamino) methyl) benzoate of formula 7-4; 0.040g, 0.16mmol) was dissolved in dimethylformamide (3mL) and potassium carbonate (0.046g, 0.33mmol) was then added slowly. 4-nitrophenyl chloroformate (0.037g, 0.18mmol) was then added, the temperature was slowly raised to 50 ℃ and the mixture was stirred for 2 days. After completion of the reaction, the ethyl acetate layer was washed three times with a saturated aqueous ammonium chloride solution, and then the organic layer was dehydrated with sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) to give the desired compound of formula 7-5 (0.048g, 71%) as a yellow oil.
1H NMR(400MHz，CDCl3)δ8.49-8.48(m，1H)，8.24(dd，2H，J＝7.0，2.2Hz)，8.17(dd，2H，J＝7.2，2.0Hz)，8.00(d，2H，J＝8.4Hz)，7.78(t，1H，J＝3.8Hz)，7.44(d，2H，J＝8.0Hz)，6.91(dd，2H，J＝7.3，2.1Hz)，5.39(brs，2H)，3.92(s，3H)；MS(ESI)m/z 408(M++H)
(formula 7-6: 4- ((N- (pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate of formula 7-5; 0.040g, 0.098mmol) was dissolved in dimethylformamide (5mL), followed by the slow addition of potassium carbonate (0.040g, 0.30mmol) and morpholine (0.013mL, 0.15mmol), after which the temperature was slowly raised to 80 ℃ and the mixture was stirred for 3 hours. The reaction was completed, and the organic layer was washed three times with saturated ammonium chloride, dehydrated with sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 50%) and concentrated to give the desired compound of formulae 7 to 6 (0.022g, 63%) as a pale yellow solid.
1H NMR(400MHz，CDCl3)δ8.37-8.35(m，1H)，7.95(d，2H，J＝8.4Hz)，7.60-7.58(m，1H)，7.47(d，2H，J＝8.4Hz)，6.94-6.89(m，2H)，5.13(s，2H)，3.89(s，3H)，3.53-3.51(m，4H)，3.31-3.29(m，4H)
(Compound 280: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) morpholine-4-carboxamide)
A compound of formula 7-6 (methyl 4- ((N- (pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoate; 0.022g, 0.062mmol) was dissolved in methanol (2mL) and hydroxylamine hydrochloride (0.022g, 0.31mmol) was slowly added. Then, potassium hydroxide (0.035g, 0.62mmol) was added and stirred at room temperature for about 10 minutes, then hydroxylamine (50.0 wt% aqueous solution; 0.082mL, 1.24mmol) was added and stirred at room temperature for 1 day. Then, the organic solvent was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. Then, the organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered to obtain the desired compound 280(0.007g, 32%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.32(d，1H，J＝3.6Hz)，7.72(t，1H，J＝6.6Hz)，7.67(d，2H，J＝8.2Hz)，7.48(d，2H，J＝8.2Hz)，7.08-7.01(m，2H)，5.08(s，2H)，3.52(t，4H，J＝4.8Hz)，3.29(t，4H，J＝4.8Hz)；MS(ESI)m/z 357(M++H)。
Example 12: synthesis of Compound 281
(formula 7-4: 4- ((pyridin-3-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 7-3 (pyridin-3-amine, 0.5g, 5.31mmol) was dissolved in methanol (10mL), and then methyl 4-formylbenzoate (1.03g, 5.31mmol) was added and stirred at room temperature for 20 minutes. Then, sodium cyanoborohydride (0.33g, 5.31mmol) and acetic acid (0.32mL, 5.31mmol) were added slowly and stirred at room temperature for 5 hours. The organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0-30%) to give the desired compound of formula 7-4 (0.85g, 66%) as a white solid.
(formula 7-5: 4- ((((4-nitrophenoxy) carbonyl) (pyridin-3-yl) amino) methyl) benzoic acid methyl ester)
The compound of formula 7-4 (methyl 4- ((pyridin-3-ylamino) methyl) benzoate; 0.20g, 0.83mmol) was dissolved in dimethylformamide (3mL), followed by the addition of 4-nitrophenyl chloroformate (0.18g, 0.90mmol) and potassium carbonate (0.23g, 1.65 mmol). Then, the temperature was slowly raised to 90 ℃, and the mixture was stirred for 1 day. After completion of the reaction, the ethyl acetate layer was washed three times with a saturated aqueous ammonium chloride solution, and then the organic layer was dehydrated with sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 100%) to give the desired compound of formula 7-5 (0.030g, 8%) as a white solid.
(formula 7-6: 4- ((N- (pyridin-3-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
The compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (pyridin-3-yl) amino) methyl) benzoate; 0.025g, 0.061mmol) was dissolved in dimethylformamide (1mL) and potassium carbonate (0.025g, 0.184mmol) and morpholine (0.008g, 0.092mmol) were added. Then, the temperature was slowly raised to 60 ℃, and the mixture was stirred for 3 hours. After completion of the reaction, the ethyl acetate layer was washed three times with a saturated aqueous ammonium chloride solution, and then the organic layer was dehydrated with sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure, and then the concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 0-10%) to give the desired compound of formula 7-6 (0.018g, 83%) as a pale yellow solid.
(Compound 281: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-3-yl) morpholine-4-carboxamide)
The compound of formula 7-6 (methyl 4- ((N- (pyridin-3-yl) morpholine-4-carboxamido) methyl) benzoate; 0.018g, 0.051mmol) was dissolved in methanol (5mL) and hydroxylamine hydrochloride (0.018g, 0.25mmol) was added slowly. Then, potassium hydroxide (0.028g, 0.51mmol) was added and stirred at room temperature for about 10 minutes, and then hydroxylamine (50.0 wt% aqueous solution; 0.067mL, 1.0mmol) was added and stirred at room temperature for 1 day. Then, the organic layer was concentrated under reduced pressure, followed by neutralization by addition of 2N hydrochloric acid. Then, the organic layer was washed three times with saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, and filtered to obtain the desired compound 281(0.010g, 55%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.11(s，1H)，8.06(d，1H，J＝4.1Hz)，8.03(d，2H，J＝8.3Hz)，7.42(d，2H，J＝2.8Hz)，7.27-7.21(m，2H)，4.93(s，2H)，3.51(t，4H，J＝4.7Hz)，3.25(t，4H，J＝4.8Hz)；MS(ESI)m/z 357(M++H)。
Example 13: synthesis of Compound 309
(formula 7-4: 4- ((pyrimidin-2-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 7-1 (2-chloropyrimidine; 6.25g, 54.6mmol) and the compound of formula 7-2 (methyl 4- (aminomethyl) benzoate hydrochloride; 10.0g, 49.6mmol) were dissolved in ethanol (150mL), and triethylamine (17.0mL, 124mmol) was added and stirred under reflux for 3 days. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 7-4 (8.06g, 67%) as a white solid.
(formula 7-5: 4- ((((4-nitrophenoxy) carbonyl) (pyrimidin-2-yl) amino) methyl) benzoic acid methyl ester)
The compound of formula 7-4(4- ((pyrimidin-2-ylamino) methyl) benzoic acid methyl ester; 8.06g, 33.1mmol) and 4-nitrophenyl chloroformate (7.01g, 34.8mmol) were dissolved in acetonitrile (150mL), followed by addition of potassium carbonate (6.87g, 49.7mmol) and stirring at room temperature for 16 hours. Then, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 7-5 (1.50g, 11%) as a pale yellow liquid.
(formula 7-6: 4- ((4-benzyl-N- (pyrimidin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
The compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (pyrimidin-2-yl) amino) methyl) benzoate; 0.150g, 0.367mmol) was dissolved in dimethylformamide (1mL), and then 1-benzylpiperazine (0.130g, 0.735mmol) and potassium carbonate (0.152g, 1.10mmol) were added and stirred at 60 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formulae 7 to 6 (0.132g, 81%) as a yellow liquid.
(Compound 309: 4-benzyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (pyrimidin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4-benzyl-N- (pyrimidin-2-yl) piperazine-1-carboxamido) methyl) benzoate of formula 7-6 (0.132g, 0.296mmol) was dissolved in methanol (5mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.906mL, 14.8mmol) and potassium hydroxide (0.166g, 2.96mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and a saturated aqueous sodium bicarbonate solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 309(0.079g, 60%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.18(s，1H)，9.03(s，1H)，8.50(d，2H，J＝4.8Hz)，7.68(d，2H，J＝8.4Hz)，7.39(d，2H，J＝8.4Hz)，7.32-7.21(m，5H)6.93(t，1H，J＝4.8Hz)，4.97(s，2H)，3.40(s，2H)，3.29(m，4H)，2.19(m，4H)。MS(ESI)m/z 447(M++H)。
Example 14: synthesis of Compound 311
(formula 7-6: 4- ((4-hydroxy-4-phenyl-N- (pyrimidin-2-yl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (pyrimidin-2-yl) amino) methyl) benzoate, a compound of formula 7-5 (0.150 g, 0.367mmol) was dissolved in dimethylformamide (1mL), and then 4-phenylpiperidin-4-ol (0.130g, 0.735mmol) and potassium carbonate (0.152g, 1.10mmol) were added and stirred at 60 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to obtain the desired compound of formulae 7 to 6 (0.104g, 63%) as a pale yellow liquid.
(Compound 311: 4-hydroxy-N- (4- (hydroxycarbamoyl) benzyl) -4-phenyl-N- (pyrimidin-2-yl) piperidine-1-carboxamide)
Methyl 4- ((4-hydroxy-4-phenyl-N- (pyrimidin-2-yl) piperidine-1-carboxamido) methyl) benzoate, a compound of formula 7-6 (0.104g, 0.233mmol) was dissolved in methanol (5mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.43mL, 23.3mmol) and potassium hydroxide (0.131g, 2.33mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and a saturated aqueous sodium bicarbonate solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 311(0.072g, 69%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.19(s，1H)，9.04(s，1H)，8.54(d，2H，J＝4.8Hz)，7.73(d，2H，J＝8.3Hz)，7.45(d，2H，J＝8.3Hz)，7.30-7.26(m，4H)，7.19(m，1H)，6.93(t，1H，J＝4.8Hz)，5.08(s，1H)，5.01(brs，2H)，3.73(brs，2H)，3.18-3.11(m，2H)，1.56-1.47(m，4H)。MS(ESI)m/z 448(M++H)。
Example 15: synthesis of Compound 312
(formula 7-6: 4- ((N- (pyrimidin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
The compound of formula 7-4(4- ((pyrimidin-2-ylamino) methyl) benzoic acid methyl ester; 0.200g, 0.822mmol) was dissolved in acetonitrile (2mL), then morpholine-4-carbonyl chloride (0.185g, 1.23mmol) and N, N-diisopropylethylamine (0.291mL, 1.64mmol) were added and stirred at 60 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to obtain the desired compound of formulae 7-6 (0.179g, 61%) as a pale yellow solid.
(Compound 312: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyrimidin-2-yl) morpholine-4-carboxamide)
The compound of formula 7-6(4- ((N- (pyrimidin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.179g, 0.502mmol) was dissolved in methanol (5mL), followed by the sequential addition of hydroxylamine (50.0 wt.% aqueous solution; 2.15mL, 35.2mmol) and potassium hydroxide (0.282g, 5.02mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and a saturated aqueous sodium bicarbonate solution. The organic layer was washed three times with water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was recrystallized from diethyl ether and hexane to give the desired compound 312(0.036g, 20%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.18(s，1H)，8.53(d，2H，J＝4.8Hz)，7.68(d，2H，J＝7.6Hz)，7.40(d，2H，J＝8.1Hz)，6.96(t，1H，J＝4.8Hz)，4.98(s，2H)，3.40(m，4H)，3.28(m，4H)。MS(ESI)m/z 358(M++H)。
Example 16: synthesis of Compound 313
(formula 7-6: 4- ((2, 6-dimethyl-N- (pyrimidin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (pyrimidin-2-yl) amino) methyl) benzoate, a compound of formula 7-5 (0.150 g, 0.367mmol) was dissolved in dimethylformamide (1mL), followed by addition of 2, 6-dimethylmorpholine (0.085g, 0.735mmol) and potassium carbonate (0.152g, 1.10mmol) and stirring at 60 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formulae 7-6 (0.062g, 44%) as a yellow liquid.
(Compound 313: N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (pyrimidin-2-yl) morpholine-4-carboxamide)
Methyl 4- ((2, 6-dimethyl-N- (pyrimidin-2-yl) morpholine-4-carboxamido) methyl) benzoate, 0.062g, 0.161mmol, a compound of formula 7-6 was dissolved in methanol (5mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.986mL, 16.1mmol) and potassium hydroxide (0.091g, 1.61mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and a saturated aqueous sodium bicarbonate solution. The organic layer was washed three times with water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was recrystallized from diethyl ether and hexane to give the desired compound 313(0.037g, 60%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.18(s，1H)，9.02(s，1H)，8.52(d，2H，J＝4.4Hz)，7.68(d，2H，J＝7.8Hz)，7.40(d，2H，J＝7.8Hz)，6.95(t，1H，J＝4.6Hz)，4.98(s，2H)，3.68(brs，2H)，3.21(brs，2H)，2.44-2.41(m，2H)，0.97(s，3H)，0.96(s，3H)。MS(ESI)m/z 386(M++H)。
Example 17: synthesis of Compound 329
(formula 8-2: 4- ((phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 8-1 (methyl 4- (bromomethyl) benzoate; 12.5g, 54.8mmol) and aniline (5.00mL, 54.8mmol) were dissolved in acetonitrile (70mL), then N, N-diisopropylethylamine (11.6mL, 65.7mmol) was added and stirred at room temperature for 16 h. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 15%) and concentrated to give the desired compound of formula 8-2 (9.13g, 69%) as a yellow liquid.
(formula 8-3: 4- ((4-bromophenylamino) methyl) benzoic acid methyl ester)
The compound of formula 8-2 (methyl 4- ((phenylamino) methyl) benzoate; 9.13g, 37.8mmol) was dissolved in dichloromethane (70mL) and N-bromosuccinimide (7.07g, 39.7mmol) was added and stirred at room temperature for 16 hours. After completion of the reaction, the organic layer was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 15%) and concentrated to obtain the desired compound of formula 8-3 (9.57g, 79%) as a white solid.
(formula 8-4: 4- (((4-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 8-3 (4- ((4-bromophenylamino) methyl) benzoic acid methyl ester; 7.67g, 24.0mmol) and 4-nitrophenyl chloroformate (5.31g, 26.4mmol) were dissolved in acetonitrile (70mL), followed by addition of potassium carbonate (4.97g, 35.9mmol) and stirring at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 8-4 (11.2g, 96%) as a yellow liquid.
(formula 8-5: 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
The compound of formula 8-4 (methyl 4- (((4-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 5.00g, 10.3mmol) was dissolved in dimethylformamide (20mL), and then morpholine (1.35g, 15.5mmol) and potassium carbonate (2.85g, 20.6mmol) were added and stirred at 60 ℃ for 5 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 8-5 (3.86g, 87%) as a yellow solid.
(formula 8-6: 4- ((N- (4- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.185g, 0.427mmol), 1-methyl-indazol-6-ylboronic acid (0.090g, 0.512mmol) and Pd (dppf) Cl2(0.035g, 0.043mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.181g, 1.71mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 80 ℃ for 16 h. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to obtain the desired compound of formulae 8 to 6 (0.112g, 71%) as a white solid.
(Compound 329: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (1-methyl-1H-indazol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.112g, 0.231mmol, a compound of formula 8-6 was dissolved in methanol (5mL), and hydroxylamine (50.0 wt% aqueous solution; 1.41mL, 23.1mmol) and potassium hydroxide (0.130g, 2.31mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 329(0.096g, 86%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ8.02(s，1H)，7.88(s，1H)，7.77(d，1H，J＝8.4Hz)，7.73(d，2H，J＝8.6Hz)，7.66(d，2H，J＝8.2Hz)，7.42-7.39(m，3H)，7.26(d，2H，J＝8.5Hz)，4.93(s，2H)，4.07(s，3H)，3.47-3.43(m，4H)，3.22-3.17(m，4H)。MS(ESI)m/z 486(M++H)。
Example 18: synthesis of Compound 330
(formula 8-6: 4- ((N- (biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), phenylboronic acid (0.068g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane 60%)Purification and concentration gave the desired compound of formula 8-6 (0.160g, 81%) as a white solid.
(Compound 330: N- (Biphenyl-4-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
The compound of formula 8-6(4- ((N- (biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.160g, 0.372mmol) was dissolved in methanol (10mL), followed by the sequential addition of hydroxylamine (50.0 wt% aqueous solution; 2.27mL, 37.2mmol) and potassium hydroxide (0.209g, 3.72mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 330(0.120g, 75%) as a pale brown solid.
MS(ESI)m/z 432(M++H)。
Example 19: synthesis of Compound 331
(formula 8-6: 4- ((N- (3 ', 5' -difluorobiphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 3 '5' -difluorophenylboronic acid (0.088g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), howeverThen sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 60%) and concentrated to obtain the desired compound of formulae 8-6 (0.169g, 79%) as a white solid.
(Compound 331: N- (3 ', 5' -difluorobiphenyl-4-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3 ', 5' -difluorobiphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate, 0.169g, 0.362mmol, a compound of formula 8-6 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.22mL, 36.2mmol) and potassium hydroxide (0.203g, 3.62mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 331(0.102g, 60%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.70(d，2H，J＝8.8Hz)，7.65(d，2H，J＝8.3Hz)，7.43-7.37(m，4H)，7.22(d，2H，J＝8.8Hz)，7.17(m，1H)，4.92(s，2H)，3.44(t，4H，J＝4.6Hz)，3.18(t，4H，J＝4.6Hz).MS(ESI)m/z 468(M++H)。
Example 20: synthesis of Compound 332
(formula 8-6: 4- ((N- (4- (pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), pyridin-3-ylboronic acid (0.068g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 75%) and concentrated to obtain the desired compound of formula 8-6 (0.132g, 66%) as a white solid.
(Compound 332: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (pyridin-3-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.132g, 0.306mmol) of the compound of formula 8-6 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.87mL, 30.6mmol) and potassium hydroxide (0.172g, 3.06mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 332(0.051g, 39%) as a brown solid.
1H NMR(400MHz，DMSO-d6)δ11.15(s，1H)，9.00(s，1H)，8.86(s，1H)，8.52(s，1H)，8.04(d，1H，J＝5.2Hz)，7.71-7.65(m，4H)，7.43-7.38(m，3H)，7.26(d，2H，J＝6.8Hz)，4.93(s，2H)，3.44(m，4H)，3.19(m，4H)。MS(ESI)m/z 433(M++H)。
Example 21: synthesis of Compound 333
(formula 8-6: 4- ((N- (4- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), pyrimidin-5-ylboronic acid (0.069g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 75%) and concentrated to obtain the desired compound of formulae 8-6 (0.148g, 74%) as a white solid.
(Compound 333: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.148g, 0.342mmol, a compound of formula 8-6 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.09mL, 34.2mmol) and potassium hydroxide (0.192g, 3.42mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and the organic layer was extracted with ethyl acetate and a saturated aqueous sodium bicarbonate solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was recrystallized from dichloromethane and hexane to give the desired compound 333(0.060g, 40%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ9.13(s，1H)，9.10(s，2H)，7.77(d，2H，J＝8.7Hz)，7.65(d，2H，J＝8.3Hz)，7.39(d，2H，J＝8.2Hz)，7.29(d，2H，J＝8.7Hz)，4.94(s，2H)，3.45(t，4H，J＝4.6Hz)，3.19(t，4H，J＝4.6Hz).
MS(ESI)m/z 434(M++H)。
Example 22: synthesis of Compound 334
(formula 8-6: 4- ((N- (4- (quinolin-7-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), quinolin-7-ylboronic acid (0.096g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to give the desired formula8-6 (0.192g, 86%) as a light brown solid.
(Compound 334: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (quinolin-7-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (quinolin-7-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.192g, 0.399mmol) of the compound of formula 8-6 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.44mL, 39.9mmol) and potassium hydroxide (0.224g, 3.99mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 334(0.173g, 90%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ8.89(m，1H)，8.41(dd，1H，J＝8.3，1.7Hz)，7.96(m，1H)，7.74(m，1H)，7.71-7.63(m，5H)，7.55(dd，1H，J＝8.2，4.1Hz)，7.43(d，2H，J＝8.2Hz)，7.25(d，2H，J＝8.6Hz)，4.95(s，2H)，3.48-3.45(m，4H)，3.24-3.20(m，4H)。MS(ESI)m/z 483(M++H)。
Example 23: synthesis of Compound 335
(formula 8-6: 4- ((N- (4- (biphenyl-3-yl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5(4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.200g0.462mmol), biphenyl-3-ylboronic acid (0.110g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to obtain the desired compound of formula 8-6 (0.230g, 98%) as a white solid.
(Compound 335: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (biphenyl-3-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (biphenyl-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.230g, 0.454mmol) of the compound of formula 8-6 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.78mL, 45.4mmol) and potassium hydroxide (0.255g, 4.54mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 335(0.220g, 96%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.86(s，1H)，7.75-7.70(m，4H)，7.67(d，2H，J＝8.3Hz)，7.63-7.60(m，2H)，7.53-7.45(m，3H)，7.41-7.39(m，3H)，7.24(d，2H，J＝8.6Hz)，4.92(s，2H)，3.44-3.42(m，4H)，3.19-3.17(m，4H)。MS(ESI)m/z 508(M++H)。
Example 24: synthesis of Compound 336
(formula 8-6: 4- ((N- (4- (1H-indol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl (4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), indol-5-ylboronic acid (0.089g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to obtain the desired compound of formulae 8-6 (0.191g, 88%) as a pale brown solid.
(Compound 336: N- (4- (1H-indol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (1H-indol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 8-6 (0.191g, 0.407mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.49mL, 40.7mmol) and potassium hydroxide (0.228g, 4.07mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 336(0.185g, 97%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(s，1H)，7.77(s，1H)，7.66(d，2H，J＝8.2Hz)，7.61(d，2H，J＝8.6Hz)，7.44-7.38(m，3H)，7.36-7.33(m，2H)，7.19(d，2H，J＝8.6Hz)，6.44(s，1H)，4.90(s，2H)，3.45-3.41(m，4H)，3.21-3.17(m，4H)。MS(ESI)m/z 471(M++H)。
Example 25: synthesis of Compound 337
(formula 8-6: 4- ((N- (4- (1H-indol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), indol-6-ylboronic acid (0.089g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to obtain the desired compound of formulae 8-6 (0.201g, 93%) as a pale brown solid.
(Compound 337: N- (4- (1H-indol-6-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (1H-indol-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 8-6 (0.201g, 0.428mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.62mL, 42.8mmol) and potassium hydroxide (0.240g, 4.28mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 337(0.195g, 97%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ11.15(s，1H)，7.67(d，2H，J＝8.2Hz)，7.63-7.56(m，4H)，7.39(d，2H，J＝8.2Hz)，7.36(m，1H)，7.26(m，1H)，7.21(d，2H，J＝8.6Hz)，6.42(s，1H)，4.91(s，2H)，3.44-3.42(m，4H)，3.19-3.17(m，4H)。MS(ESI)m/z 471(M++H)。
Example 26: synthesis of Compound 338
(formula 8-6: 4- ((N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-4-yl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), 3, 4, 5-trimethoxyphenylboronic acid (0.117g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL) then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reactionThe solution was stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to obtain the desired compound of formula 8-6 (0.121g, 50%) as a white solid.
(Compound 338: N- (4- (hydroxycarbamoyl) benzyl) -N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-4-yl) morpholine-4-carboxamide)
Reacting methyl (4- ((N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate of formula 8-6; 0.121g, 0.232mmol) was dissolved in methanol (10mL) and then hydroxylamine (50.0 wt% aqueous solution; 1.42mL, 23.2mmol) and potassium hydroxide (0.130g, 2.32mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 338(0.038g, 31%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.66-7.61(m，4H)，7.38(d，2H，J＝8.2Hz)，7.20(d，2H，J＝8.6Hz)，6.86(s，2H)，4.91(s，2H)，3.83(s，6H)，3.66(s，3H)，3.44(t，4H，J＝4.3Hz)，3.17(t，4H，J＝4.4Hz).MS(ESI)m/z522(M++H)。
Example 27: synthesis of Compound 339
(formula 8-6: 4- ((N- (3 ', 5' -bis (trifluoromethyl) biphenyl-4-yl)) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 3, 5-bis (trifluoromethyl) phenylboronic acid (0.143g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formula 8-6 (0.203g, 78%) as a white solid.
(Compound 339: N- (3 ', 5' -bis (trifluoromethyl) biphenyl-4-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Reacting a compound of formula 8-6 (methyl 4- ((N- (3 ', 5' -bis (trifluoromethyl) biphenyl-4-yl)) morpholine-4-carboxamido) methyl) benzoate; 0.203g, 0.358mmol) was dissolved in methanol (10mL) and then hydroxylamine (50.0 wt% aqueous solution; 2.19mL, 35.8mmol) and potassium hydroxide (0.201g, 3.58mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 339(0.197g, 97%) as a light brown solid.
1H NMR(400MHz，DMSO-d6)δ8.29(s，2H)，8.03(s，1H)，7.82(d，2H，J＝8.6Hz)，7.66(d，2H，J＝8.2Hz)，7.40(d，2H，J＝8.2Hz)，7.28(d，2H，J＝8.6Hz)，4.94(s，2H)，3.47-3.43(m，4H)，3.21-3.15(m，4H)。MS(ESI)m/z 568(M++H)。
Example 28: synthesis of Compound 340
(formula 8-6: 4- ((N- (4- (1H-indol-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), indol-4-ylboronic acid (0.089g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then sodium carbonate (0.196g, 1.85mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 6 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to obtain the desired compound of formula 8-6 (0.142g, 66%) as a pale yellow solid.
(Compound 340: N- (4- (1H-indol-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (1H-indol-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 8-6 (0.142g, 0.302mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.85mL, 30.2mmol) and potassium hydroxide (0.170g, 3.02mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 3mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 340(0.105g, 74%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.25(s，1H)，7.68(d，2H，J＝8.3Hz)，7.61(d，2H，J＝8.6Hz)，7.41-7.36(m，4H)，7.25(d，2H，J＝8.6Hz)，7.14(t，1H，J＝7.7Hz)，7.04(d，1H，J＝7.3Hz)，6.51(s，1H)，4.92(s，2H)，3.45-3.43(m，4H)，3.22-3.20(m，4H)。MS(ESI)m/z 471(M++H)。
Example 29: synthesis of Compound 341
(formula 7-4: 4- ((pyridin-2-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 7-3 (pyridin-2-amine; 10.0g, 106mmol) was dissolved in methanol (100mL), followed by addition of methyl 4-formylbenzoate (17.4g, 106mmol) and acetic acid (6.08mL, 106mmol) and stirring for 1 day. Then, sodium cyanoborohydride (22.5g, 106mmol) was added and stirred for 1 day. After completion of the reaction, the resultant solid was filtered, and the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; dichloromethane/ethyl acetate ═ 5%) and concentrated to give the desired compound of formula 7-4 (8.0g, 31%) as a white solid.
(formula 7-5: 4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoic acid methyl ester)
The compound of formula 7-4 (methyl 4- ((pyridin-2-ylamino) methyl) benzoate; 1.80g, 7.43mmol) was dissolved in acetonitrile (50mL), followed by the addition of potassium carbonate (3.08g, 22.3mmol) and 4-nitrophenyl chloroformate (2.24g, 11.1 mmol). The mixture was then heated and stirred at about 50 ℃ for 1 day. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; hexane/ethyl acetate ═ 30%) and concentrated to give the desired compound of formula 7-5 (0.98, 32%) as a white solid.
(formula 7-6: 4- ((N- (pyridine-2-yl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.30g, 0.736mmol) was dissolved in dimethylformamide (4mL), followed by the addition of piperidine (0.06mL, 0.736mmol) and potassium carbonate (0.122g, 0.884mmol), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.27g, 104%) as a yellow oil.
(Compound 341: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperidine-1-carboxamide)
Methyl 4- ((N- (pyridin-2-yl) piperidine-1-carboxamido) methyl) benzoate of formula 7-6; 0.13g, 0.38mmol) was dissolved in methanol (10mL), and hydroxylamine (0.13g, 1.89mmol) and potassium hydroxide (0.21g, 3.79mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 0.98mL, 7.58mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate, washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 341(0.09g, 70%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.31-8.29(m，1H)，7.72-7.66(m，3H)，7.49(d，2H，J＝8.3Hz)，7.01-6.98(m，2H)，5.07(s，2H)，3.27(t，4H，J＝5.5Hz)，1.57-1.53(m，2H)，1.45-1.41(m，4H)；MS(ESI)m/z 355.2(M++H)。
Example 30: synthesis of Compound 342
(formula 7-6: 4- ((4-methyl-N- (pyridine-2-yl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.300g, 0.736mmol) was dissolved in dimethylformamide (4mL), then 4-methylpiperidine (0.07g, 0.736mmol) and potassium carbonate (0.122g, 0.884mmol) were added and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.27g, 100%) as a yellow oil.
(Compound 342: N- (4- (hydroxycarbamoyl) benzyl) -4-methyl-N- (pyridin-2-yl) piperidine-1-carboxamide)
Methyl 4- ((4-methyl-N- (pyridin-2-yl) piperidine-1-carboxamido) methyl) benzoate, 0.15g, 0.40mmol, a compound of formula 7-6 was dissolved in methanol (10mL), and hydroxylamine (0.14g, 2.00mmol) and potassium hydroxide (0.22g, 4.00mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 1.03mL, 8.00mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate, washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 342(0.09g, 64%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.31-8.29(m，1H)，7.71-7.67(m，3H)，7.48(d，2H，J＝8.3Hz)，7.01-6.98(m，2H)，5.07(s，2H)，3.83(d，2H，J＝13.2Hz)，2.72(td，2H，J＝12.6，2.0Hz)，1.55-1.50(m，3H)，0.97-0.93(m，2H)，0.95(m，3H)；MS(ESI)m/z 369.1(M++H)。
Example 31: synthesis of Compound 343
(formula 7-6: 4- ((2, 6-dimethyl-N- (pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.300g, 0.736mmol) was dissolved in dimethylformamide (4mL), then 2, 6-dimethylmorpholine (0.085g, 0.736mmol) and potassium carbonate (0.122g, 0.884mmol) were added and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.17g, 60%) as a white oil.
(Compound 343: N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (pyridin-2-yl) morpholine-4-carboxamide)
Methyl 4- ((2, 6-dimethyl-N- (pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoate, 0.116g, 0.303mmol, a compound of formula 7-6 was dissolved in methanol (10mL), and hydroxylamine (0.105g, 1.51mmol) and potassium hydroxide (0.169g, 3.02mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 0.778mL, 6.05mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate, washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 343(0.107g, 92%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.32(dd，1H，J＝4.9，1.1Hz)，7.74-7.69(m，1H)，7.67(d，2H，J＝8.2Hz)，7.48(d，2H，J＝8.2Hz)，7.05-7.02(m，2H)，5.08(s，2H)，3.65(d，2H，J＝12.9Hz)，3.43-3.37(m，2H)，2.46-2.40(m，2H)，1.03(d，6H，J＝6.2Hz)；MS(ESI)m/z 385.1(M++H)。
Example 32: synthesis of Compound 352
(formula 7-6: 4- ((4-phenyl-N- (pyridin-2-yl) -1, 2, 3, 6-tetrahydropyridine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate of formula 7-5; 0.50g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 4-phenyl-1, 2, 3, 6-tetrahydropyridine (0.264g, 1.35mmol) and potassium carbonate (0.339g, 2.46mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formulae 7-6 (0.57g, 108%) as a yellow oil.
(Compound 352: N- (4- (hydroxycarbamoyl) benzyl) -4-phenyl-N- (pyridin-2-yl) -5, 6-dihydropyridine-1 (2H) -carboxamide)
Methyl 4- ((4-phenyl-N- (pyridin-2-yl) -1, 2, 3, 6-tetrahydropyridine-1-carboxamido) methyl) benzoate, 0.386g, 0.903mmol, of the compound of formula 7-6 was dissolved in methanol (20mL), and hydroxylamine (0.313g, 4.51mmol) and potassium hydroxide (0.507g, 9.03mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 2.33mL, 18.1mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, methanol was evaporated under reduced pressure, and a solid was precipitated by adding 2N hydrogen chloride. The solid was filtered and dried to give the desired compound 352(0.22g, 57%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.33(dd，1H，J＝5.0，1.2Hz)，7.74-7.70(m，1H)，7.65(d，2H，J＝8.3Hz)，7.49(d，2H，J＝8.3Hz)，7.36-7.23(m，5H)，7.09-7.02(m，2H)，5.97(t，1H，J＝3.2Hz)，5.12(s，2H)，3.90(d，2H，J＝3.0Hz)，3.59(t，2H，J＝5.7Hz)，2.43(d，2H，J＝5.7Hz)；MS(ESI)m/z 429.1(M++H)。
Example 33: synthesis of Compound 353
(formula 7-6: 4- ((4-methyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.5g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 1-methylpiperazine (0.15mL, 1.35mmol) and potassium carbonate (0.339g, 2.46mmol) were added, and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formula 7-6 (0.33g, 73%) as a yellow oil.
(Compound 353: N- (4- (hydroxycarbamoyl) benzyl) -4-methyl-N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4-methyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoate, 0.23g, 0.624mmol of the compound of formula 7-6 was dissolved in methanol (20mL), and then hydroxylamine (0.217g, 3.12mmol) and potassium hydroxide (0.35g, 6.24mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 1.61mL, 12.49mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, methanol was evaporated under reduced pressure, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 353(0.04g, 17%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.33(dd，1H，J＝4.9，1.6Hz)，7.75-7.71(m，1H)，7.68(d，2H，J＝8.2Hz)，7.48(d，2H，J＝8.1Hz)，7.09(d，1H，J＝8.2Hz)，7.06-7.03(m，1H)，5.08(s，2H)，3.41-3.38(m，4H)，2.59-2.57(m，4H)，2.44(s，3H)；MS(ESI)m/z 370.1(M++H)。
Example 34: synthesis of Compound 354
(formula 7-6: 4- ((4-Ethyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 1-ethylpiperazine (0.154g, 1.35mmol) and potassium carbonate (0.339g, 2.45mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 30%) and concentrated to give the desired compound of formula 7-6 (0.3g, 64%) as a yellow oil.
(Compound 354: 4-Ethyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4-ethyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoate, 0.23g, 0.60mmol, of the compound of formula 7-6 was dissolved in methanol (20mL), and hydroxylamine (0.209g, 3.01mmol) and potassium hydroxide (0.337g, 6.01mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 1.55mL, 12.02mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 354(0.038g, 17%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.31(dd，1H，J＝4.8，1.2Hz)，7.73-7.66(m，3H)，7.48(d，2H，J＝8.2Hz)，7.06-7.01(m，2H)，5.08(s，2H)，3.36-3.31(m，4H)，2.46-2.37(m，6H)，1.07(t，3H，J＝7.2Hz)；MS(ESI)m/z 384.1(M++H)。
Example 35: synthesis of Compound 355
(formula 7-6: 4- ((4-benzyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 1-benzylpiperazine (0.231mL, 1.35mmol) and potassium carbonate (0.339g, 2.46mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formulae 7-6 (0.185g, 34%) as a yellow oil.
(Compound 355: 4-benzyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4-benzyl-N- (pyridin-2-yl) -piperazine-1-carboxamido) methyl) benzoate of formula 7-6, 0.136g, 0.31mmol, was dissolved in methanol (20mL), then hydroxylamine (0.106g, 1.53mmol) and potassium hydroxide (0.172g, 3.06mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 0.788mL, 6.12mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, methanol was evaporated under reduced pressure, and a solid was precipitated by adding 2N hydrogen chloride. The solid was filtered to give the desired compound 355(0.077g, 57%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.31(dd，1H，J＝4.8，0.9Hz)，7.72-7.65(m，3H)，7.47(d，2H，J＝8.2Hz)，7.32-7.24(m，5H)，7.03-7.00(m，2H)，5.07(s，2H)，3.48(s，2H)，3.31-3.30(m，4H)，2.32-2.29(m，4H)；MS(ESI)m/z 446.1(M++H)。
Example 36: synthesis of Compound 356
(formula 7-6: 4- ((4- (2-methoxyphenyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 1- (2-methoxyphenyl) piperazine (0.236mL, 1.35mmol) and potassium carbonate (0.339g, 2.46mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 7-6 (0.565g, 100%) as a yellow oil.
(Compound 356: N- (4- (hydroxycarbamoyl) benzyl) -4- (2-methoxyphenyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4- (2-methoxyphenyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoate, 0.395g, 0.858mmol of the compound of formula 7-6 was dissolved in methanol (20mL), and then hydroxylamine (0.298g, 4.29mmol) and potassium hydroxide (0.481g, 8.58mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 2.21mL, 17.15mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 356(0.28g, 70%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.33(dd，1H，J＝4.9，1.2Hz)，7.74-7.71(m，1H)，7.68(d，2H，J＝8.2Hz)，7.51(d，2H，J＝8.2Hz)，7.09-6.87(m，6H)，5.11(s，2H)，3.82(s，3H)，3.47-3.44(m，4H)，2.87-2.85(m，4H)；MS(ESI)m/z 446.1(M++H)。
Example 37: synthesis of Compound 357
(formula 7-6: 4- ((4- (4-fluorophenyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (3mL), then 1- (4-fluorophenyl) piperazine (0.243mL, 1.35mmol) and potassium carbonate (0.339g, 2.46mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formulae 7-6 (0.54g, 98%) as a yellow oil.
(Compound 357: 4- (4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4- (4-fluorophenyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoate, 0.31g, 0.691mmol of the compound of formula 7-6 was dissolved in methanol (20mL), and then hydroxylamine (0.24g, 3.46mmol) and potassium hydroxide (0.3881g, 6.91mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 1.78mL, 13.8mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 357(0.27g, 87%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.34-8.33(m，1H)，7.75-7.71(m，1H)，7.67(d，2H，J＝8.2Hz)，7.51(d，2H，J＝8.2Hz)，7.09(d，1H，J＝8.3Hz)，7.05-6.89(m，5H)，5.11(s，2H)，3.46-3.44(m，4H)，2.96-2.93(m，4H)；MS(ESI)m/z 450.1(M++H)。
Example 38: synthesis of Compound 358
(formula 7-6: 4- ((N- (pyridin-2-yl) pyrrolidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.20g, 0.491mmol) was dissolved in dimethylformamide (4mL), then pyrrolidine (0.035mL, 0.491mmol) and potassium carbonate (0.081g, 0.589mmol) were added, and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.031g, 19%) as a colorless oil.
(Compound 358: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) pyrrolidine-1-carboxamide)
Methyl (4- ((N- (pyridin-2-yl) pyrrolidine-1-carboxamido) methyl) benzoate of formula 7-6; 0.031g, 0.091mmol) was dissolved in methanol (4mL) and then hydroxylamine (0.032g, 0.457mmol) and potassium hydroxide (0.051g, 0.913mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 0.135mL, 1.83mmol) was added dropwise and stirred at room temperature for 12 hours. After completion of the reaction, methanol was evaporated under reduced pressure, 2N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 358(0.012g, 39%) as a colorless oil.
1H NMR(400MHz，MeOD-d3)δ8.31(dd，1H，J＝4.9，1.3Hz)，7.73-7.69(m，1H)，7.65(d，2H，J＝8.2Hz)，7.47(d，2H，J＝8.2Hz)，7.07-7.02(m，2H)，5.08(s，2H)，3.19-3.18(m，4H)，1.80-1.77(m，4H)；MS(ESI)m/z 341.2(M++H)。
Example 39: synthesis of Compound 370
(formula 11-3: 4- (Thiouromethyl) benzoic acid methyl ester)
The compound of formula 11-2 (1, 1' -thiocarbonyldiimidazole; 8.26g, 46.4mmol) was dissolved in hydrogen chloride (40mL), and then triethylamine (6.34mL, 46.4mmol) and the compound of formula 11-1 (4- (aminomethyl) benzoate hydrochloride; 8.50g, 42.2mmol) were successively added slowly, followed by stirring at room temperature for 3 hours. Then, ammonia (28.0 wt% aqueous solution; 30mL) and methanol (10mL) were added to the mixture and stirred at room temperature for 2 hours, followed by further addition of hexane (85mL) and water (45mL) and stirring at the same temperature for 1 hour. The solid product was filtered, washed with hexane then water, followed by vacuum drying to give the desired compound of formula 11-3 (9.40g, 99%) as a pale brown solid.
(formula 11-5: 4- ((4-phenylthiazol-2-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 11-3 (methyl 4- (thioureidomethyl) benzoate; 0.180g, 0.803mmol) and the compound of formula 11-4 (2-bromoacetophenone; 0.084g, 0.843mmol) were dissolved in ethanol (5mL) and then stirred under reflux for 16 hours. After the completion of the reaction, the temperature of the reaction solution was cooled to room temperature, and then ethyl acetate and hexane were added and stirred. The solid product was then filtered and dried in vacuo to give the desired compound of formula 11-5 (0.175g, 67%) as a pale brown solid.
(formula 11-6: 4- ((((4-nitrophenoxy) carbonyl) (4-phenylthiazol-2-yl) amino) methyl) benzoic acid methyl ester)
A compound of formula 11-5 (4- ((4-phenylthiazol-2-ylamino) methyl) benzoic acid methyl ester; 0.175g, 0.539mmol) and 4-nitrophenyl chloroformate (0.141g, 0.701mmol) were dissolved in acetonitrile (10mL), followed by addition of potassium carbonate (0.112g, 0.809mmol) and stirring at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 11-6 (0.160g, 61%) as a yellow solid.
(formula 11-7: 4- ((N- (4-phenylthiazol-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (4-phenylthiazol-2-yl) amino) methyl) benzoate, formula 11-6, 0.160g, 0.327mmol was dissolved in dimethylformamide (5mL), then morpholine (0.057g, 0.654mmol) and potassium carbonate (0.136g, 0.981mmol) were added and stirred at 60 ℃ for 16 h. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 11-7 (0.085g, 59%) as a pale yellow solid.
(Compound 370: N- (4- (hydroxycarbamoyl) benzyl) -N- (4-phenylthiazol-2-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (4-phenylthiazol-2-yl) morpholine-4-carboxamido) methyl) benzoate, 0.085g, 0.194mmol) of the compound of formula 11-7 was dissolved in methanol (5mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.19mL, 19.4mmol) and potassium hydroxide (0.109g, 1.94mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 370(0.059g, 69%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.82(d，2H，J＝8.5Hz)，7.67(d，2H，J＝8.3Hz)，7.55(s，1H)，7.41-7.35(m，4H)，7.29(m，1H)，5.13(s，2H)，3.59-3.57(m，4H)，3.39-3.36(m，4H)。MS(ESI)m/z 439(M++H)。
Example 40: synthesis of Compound 371
(formula 11-5: 4- ((4- (4-chlorophenyl) thiazol-2-ylamino) methyl) benzoic acid methyl ester)
The compound of formula 11-3 (methyl 4- (thioureidomethyl) benzoate; 0.500g, 2.23mmol) and 2-bromo-4-chloroacetophenone (0.573g, 2.45mmol) were dissolved in ethanol (20mL) and then stirred under reflux for 16 hours. After the completion of the reaction, the temperature of the reaction solution was cooled to room temperature, and then ethyl acetate and hexane were added and stirred. Then, the solid product was filtered and dried in vacuo to give the desired compound of formula 11-5 (0.700g, 88%) as a white solid.
(formula 11-6: 4- (((4- (4-chlorophenyl) thiazol-2-yl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 11-5 (4- ((4- (4-chlorophenyl) thiazol-2-ylamino) methyl) benzoate; 0.500g, 1.39mmol) and 4-nitrophenyl chloroformate (0.337g, 1.67mmol) were dissolved in dimethylformamide (10mL), followed by addition of N, N-diisopropylethylamine (0.370mL, 2.09mmol) and stirring at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 15%) and concentrated to give the desired compound of formula 11-6 (0.520g, 71%) as a yellow solid.
(formula 11-7: 4- ((N- (4- (4-chlorophenyl) thiazol-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4- (4-chlorophenyl) thiazol-2-yl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, formula 11-6, 0.520g, 0.992mmol was dissolved in dimethylformamide (5mL), then morpholine (0.173g, 1.99mmol) and potassium carbonate (0.412g, 2.98mmol) were added and stirred at 60 ℃ for 16 h. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 11-7 (0.460g, 98%) as a white solid.
(Compound 371: N- (4- (4-chlorophenyl) thiazol-2-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (4-chlorophenyl) thiazol-2-yl) morpholine-4-carboxamido) methyl) benzoate, 0.460g, 0.975mmol) of the compound of formula 11-7 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 2.98mL, 48.7mmol) and potassium hydroxide (0.547g, 9.75mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. Then, the solid product was filtered, washed with water, followed by vacuum drying to obtain the desired compound 371(0.435g, 94%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(s，1H)，9.00(s，1H)，7.83(d，2H，J＝8.5Hz)，7.67(d，2H，J＝8.2Hz)，7.62(s，1H)，7.45(d，2H，J＝8.6Hz)，7.39(d，2H，J＝8.0Hz)，5.14(s，2H)，3.60-3.57(m，4H)，3.41-3.38(m，4H)。MS(ESI)m/z 473(M++H)。
Example 41: synthesis of Compound 372
(Compound 372: N- (4-bromophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
A compound of formula 8-5(4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.085g, 0.196mmol) was dissolved in methanol (5mL), followed by the sequential addition of hydroxylamine (50.0 wt.% aqueous solution; 1.20mL, 19.6mmol) and potassium hydroxide (0.110g, 1.96mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 372(0.049g, 58%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.4Hz)，7.45(dd，2H，J＝6.8，2.1Hz)，7.33(d，2H，J＝8.2Hz)，7.09(dd，2H，J＝6.8，2.1Hz)，4.85(s，2H)，3.41(t，4H，J＝4.7Hz)，3.13(t，4H，J＝4.6Hz).MS(ESI)m/z 434，436(M++H)。
Example 42: synthesis of Compound 374
(formula 1-2: 4- ((3- (trifluoromethyl) phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (3- (trifluoromethyl) aniline; 0.30g, 1.84mmol) and potassium carbonate (0.76g, 5.53mmol) were dissolved in dimethylformamide (DMF, 5mL), followed by the addition of methyl 4- (bromomethyl) benzoate (0.42g, 1.84 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formula 1-2 (0.37g, 65%).
1H NMR(400MHz，DMSO-d6)δ7.93(d，2H，J＝8.3Hz)，7.49(d，2H，J＝8.3Hz)，7.24(t，1H，J＝7.9Hz)，6.88-6.78(m，4H)，4.42(d，2H，J＝6.1Hz)，3.83(s，3H)，MS(ESI)m/z 310(M++H)。
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((3- (trifluoromethyl) phenylamino) methyl) benzoate; 0.26g, 0.82mmol) and 4-nitrophenyl chloroformate (0.33g, 1.65mmol) were dissolved in acetonitrile (10mL), followed by addition of potassium carbonate (0.34g, 2.47 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formulae 1-3 (0.35g, 89%) as a colorless oil.
1H NMR(400MHz，CDCl3)δ8.20(d，2H，J＝10.2Hz)，8.01(d，2H，J＝7.8Hz)，7.56-7.46(m，3H)，7.35(d，3H，J＝8.0Hz)，7.26(d，2H，J＝8.1Hz)，5.01(bs，2H)，3.90(s，3H)。
(formula 1-4: 4- ((N- (3- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.29g, 0.60mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (10mL), followed by addition of potassium carbonate (0.25g, 1.81mmol) and morpholine (0.05mL, 0.60 mmol). The mixture was reacted at 60 ℃ for 2 days, and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compounds of formulae 1-4 (0.15g, 60%).
1H NMR(400MHz，DMSO-d6)δ7.97(d，2H，J＝8.2Hz)，7.43-7.32(m，5H)，7.20(d，1H，J＝8.0Hz)，4.94(s，2H)，3.90(s，3H)，3.50(t，4H，J＝4.8Hz)，3.25(t，4H，J＝4.8Hz)，MS(ESI)m/z 423(M++H)。
(Compound 374: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
The compound of formula 1-4 (methyl 4- ((N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.15g, 0.36mmol) was dissolved in methanol (5mL) and then aqueous hydroxylamine (50 wt%, 1mL) and potassium hydroxide (0.10g, 1.81mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give a solid product, and the resulting solid was filtered and dried to give the desired compound 374(0.082g, 54%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ11.14(brs，1H)，8.99(brs，1H)，7.85(d，2H，J＝8.0Hz)，7.66-7.27(m，6H)，4.94(s，2H)，3.41(s，2H)，3.15(s，2H)。MS(ESI)m/z 424(M++H)。
Example 43: synthesis of Compound 376
(formula 7-6: 4- ((4-methyl-N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (4mL), then 1-methyl-1, 4-diazepane (0.183mL, 1.47mmol) and potassium carbonate (0.339g, 2.45mmol) were added and the mixture was heated and stirred at 55 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 7-6 (0.32g, 68%) as a pale yellow oil.
(Compound 376: N- (4- (hydroxycarbamoyl) benzyl) -4-methyl-N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamide)
A compound of formula 7-6 (methyl 4- ((4-methyl-N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamido) methyl) benzoate; 0.15g, 0.39mmol) was dissolved in methanol (20mL) and then hydroxylamine (0.136g, 1.96mmol) and potassium hydroxide (0.22g, 3.92mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 1.01mL, 7.84mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was evaporated under reduced pressure, 1N hydrogen chloride was added, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 376(0.009g, 6%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.33-8.31(m，1H)，7.74-7.72(m，1H)，7.67(d，2H，J＝8.3Hz)，7.50(d，2H，J＝8.2Hz)，7.10-7.04(m，2H)，5.09(d，2H，J＝2.4Hz)，3.96-3.93(m，2H)，3.56-2.95(m，6H)，2.88(s，3H)，2.06-2.02(m，2H)；MS(ESI)m/z 384.1(M++H)。
Example 44: synthesis of Compound 377
(formula 7-6: 4- ((N- (pyridin-2-yl) azetidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (4mL), then azetidine (0.183g, 1.47mmol) and potassium carbonate (0.339g, 2.45mmol) were added and the mixture was heated and stirred at 55 ℃ for 1 day. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.03g, 8%) as a white solid.
(Compound 377: N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) azetidine-1-carboxamide)
Methyl 4- ((N- (pyridin-2-yl) azetidine-1-carboxamido) methyl) benzoate of formula 7-6, 0.03g, 0.09mmol was dissolved in methanol (20mL), and hydroxylamine (0.032g, 0.092mmol) and potassium hydroxide (0.052g, 0.922mmol) were added and stirred for 5 minutes. Then, hydroxylamine (50 wt% aqueous solution; 0.238mL, 1.84mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, 1N hydrogen chloride was added and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 377(0.013g, 43%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.38-8.36(m，1H)，7.78-7.74(m，1H)，7.64(d，2H，J＝8.3Hz)，7.40(d，2H，J＝8.3Hz)，7.24(d，1H，J＝8.2Hz)，7.15-7.11(m，1H)，5.09(s，2H)，3.73(t，4H，J＝7.7Hz)，2.12(tt，2H，J＝7.7Hz)；MS(ESI)m/z 327.2(M++H)。
Example 45: synthesis of Compound 379
(formula 7-6: 4- ((4- (3, 4-dimethylphenyl-N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.50g, 1.23mmol) was dissolved in dimethylformamide (4mL), then 1- (3, 4-dimethylphenyl) piperazine (0.28g, 1.47mmol) and potassium carbonate (0.339g, 2.46mmol) were added and the mixture was heated and stirred at 55 ℃ for 1 day. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 7-6 (0.4g, 71%) as a yellow oil.
(Compound 379: 4- (3, 4-dimethylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4- (3, 4-dimethylphenyl-N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoate, 0.1g, 0.218mmol, a compound of formula 7-6(4- ((4- (3, 4-dimethylphenyl-N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) was dissolved in methanol (20mL), then hydroxylamine (0.076g, 1.09mmol) and potassium hydroxide (0.122g, 2.18mmol) were added and stirred for 5 minutes, then hydroxylamine (50 wt% aqueous solution; 0.56mL, 4.36mmol) was added dropwise and stirred at room temperature for 6 hours, after completion of the reaction, 2N hydrogen chloride was added and the organic layer was extracted with ethyl acetate, the organic layer was washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, then concentrated under reduced pressure, the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound 379(0.05g, 52%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ8.33(dd，1H，J＝4.9，1.2Hz)，7.74-7.70(m，1H)，7.67(d，2H，J＝8.3Hz)，7.50(d，2H，J＝8.3Hz)，7.08(d，1H，J＝8.3Hz)，7.05-7.02(m，1H)，6.97(d，1H，J＝8.3Hz)，6.72(d，1H，J＝2.2Hz)，6.63(dd，1H，J＝8.2，2.4Hz)，5.10(s，2H)，3.45-3.43(m，4H)，2.95-2.93(m，4H)，2.20(s，3H)，2.15(s，3H)；MS(ESI)m/z 460.2(M++H)。
Example 46: synthesis of Compound 380
(formula 9-1: 4- ((N- (4' -formylbiphenyl-4-yl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 8-5 (methyl 4- ((N- (4-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 1.00g, 2.31mmol), 4-formylphenylboronic acid (0.415g, 2.77mmol) and Pd (dppf) Cl2(0.094g, 0.115mmol) was dissolved in 1, 4-bisAlkane (12mL), then cesium carbonate (2.24g, 6.92mmol) dissolved in water (3mL) was added to the reaction solution, followed by stirring in a microwave reactor at 140 ℃ for 15 minutes. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 9-1 (0.750g, 71%) as a pale yellow solid.
(formula 9-2: 4- ((N- (4' - (morpholinomethyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 9-1 (methyl 4- ((N- (4' -formylbiphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate; 0.150g, 0.327mmol) was dissolved in methanol (5mL), followed by addition of morpholine (0.086g, 0.981mmol) and acetic acid (0.094mL, 1.64mmol) and stirring at room temperature for 1 hour. Then, sodium cyanoborohydride (0.041g, 0.654mmol) was added and stirred at the same temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered, and then the filtrate was concentrated under reduced pressure. Then, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formula 9-2 (0.136g, 79%) as a white solid.
(Compound 380: N- (4- (hydroxycarbamoyl) benzyl) -N- (4' - (morpholinomethyl) biphenyl-4-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (4' - (morpholinomethyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 9-2 (0.130g, 0.245mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.50mL, 24.5mmol) and potassium hydroxide (0.138g, 2.46mmol) were added, and the mixture was stirred at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was recrystallized from dichloromethane and hexane to give the desired compound 380(0.073g, 56%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(brs，H)，9.14(brs，1H)，7.65(d，2H，J＝8.1Hz)，7.62-7.56(m，4H)，7.39-7.33(m，4H)，7.21(d，2H，J＝8.5Hz)，4.90(s，2H)，3.58-3.55(m，4H)，3.46(s，2H)，3.42-3.37(m，8H)，3.17(m，4H)。MS(ESI)m/z 529(M++H)。
Example 47: synthesis of Compound 381
(formula 9-2: 4- ((N- (4' - ((4-methylpiperazin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 9-1 (methyl 4- ((N- (4' -formylbiphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate; 0.150g, 0.327mmol) was dissolved in methanol (5mL), and then 1-methylpiperazine (0.098g, 0.981mmol) and acetic acid (0.094mL, 1.64mmol) were added and stirred at room temperature for 1 hour. Then, sodium cyanoborohydride (0.041g, 0.654mmol) was added and stirred at the same temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered, and then the filtrate was concentrated under reduced pressure. Then, the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 9-2 (0.140g, 79%) as a white solid.
(Compound 381: N- (4- (hydroxycarbamoyl) benzyl) -N- (4' - ((4-methylpiperazin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (4' - ((4-methylpiperazin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate of the formula 9-2 (0.140g, 0.258mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.58mL, 25.8mmol) and potassium hydroxide (0.145g, 2.58mmol) were added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was recrystallized from dichloromethane and hexane to give the desired compound 381(0.093g, 66%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝7.8Hz)，7.61-7.56(m，4H)，7.38(d，2H，J＝8.0Hz)，7.32(d，2H，J＝7.6Hz)，7.21(d，2H，J＝8.5Hz)，4.90(s，2H)，3.59-3.39(m，10H)，3.17(m，4H)，2.40-2.23(m，4H)，2.18(s，3H)。MS(ESI)m/z 544(M++H)。
Example 48: synthesis of Compound 382
(formula 9-2 (S) -methyl 4- ((N- (4' - ((2- (hydroxymethyl) pyrrolidin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate)
The compound of formula 9-1 (methyl 4- ((N- (4' -formylbiphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate; 0.150g, 0.327mmol) was dissolved in methanol (5mL), followed by addition of (S) -pyrrolidin-2-yl-methanol (0.099g, 0.981mmol) and acetic acid (0.094mL, 1.64mmol) and stirring at room temperature for 1 hour. Then, sodium cyanoborohydride (0.041g, 0.654mmol) was added and stirred at the same temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered, and then the filtrate was concentrated under reduced pressure. Then, the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 9-2 (0.123g, 69%) as a white solid.
(Compound 382 (S) -N- (4- (hydroxycarbamoyl) benzyl) -N- (4' - ((2- (hydroxymethyl) pyrrolidin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamide)
Methyl ((S) -4- ((N- (4' - ((2- (hydroxymethyl) pyrrolidin-1-yl) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate; 0.120g, 0.221mmol) of the compound of formula 9-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.35mL, 22.1mmol) and potassium hydroxide (0.124g, 2.21mmol) were added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain the desired compound 382(0.036g, 30%) as a colorless liquid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝7.8Hz)，7.61-7.55(m，4H)，7.39-7.34(m，4H)，7.21(d，2H，J＝8.3Hz)，4.90(s，2H)，4.06(m，1H)，3.47-3.39(m，6H)，3.37-3.35(m，2H)，3.17(m，4H)，2.24-2.20(m，2H)，1.64-1.52(m，5H)。MS(ESI)m/z 545(M++H)。
Example 49: synthesis of Compound 383
(formula 9-2: 4- ((N- (4' - (((2- (hydroxyethyl) (methyl) amino) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
The compound of formula 9-1 (4- ((N- (4' -formylbiphenyl-4-yl) morpholine-4-carboxamido) methyl benzoate; 0.150g, 0.327mmol) was dissolved in methanol (5mL), then 2- (methylamino) ethanol (0.074g, 0.981mmol) and acetic acid (0.094mL, 1.64mmol) were added and stirred at room temperature for 1 hour, then, sodium cyanoborohydride (0.041g, 0.654mmol) was added and stirred at the same temperature for 16 hours, after completion of the reaction, the organic layer was extracted with ethyl acetate and a saturated aqueous sodium bicarbonate solution and concentrated under reduced pressure, then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered, then the filtrate was concentrated under reduced pressure, then, the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 7%) and concentrated to give the desired compound of formula 9-2 (0.152g, 90%) as a white solid.
(Compound 383: N- (4- (hydroxycarbamoyl) benzyl) -N- (4' - ((((2-hydroxyethyl) (methyl) amino) methyl) biphenyl-4-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (4' - ((2- (hydroxyethyl) (methyl) amino) methyl) biphenyl-4-yl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 9-2 (0.150 g, 0.290mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.77mL, 29.0mmol) and potassium hydroxide (0.163g, 2.90mmol) were added, followed by stirring at room temperature for 30 minutes.
1H NMR(400MHz，DMSO-d6)δ7.66-7.56(m，6H)，7.39-7.34(m，4H)，7.21(d，2H，J＝8.6Hz)，4.90(s，2H)，3.51-3.48(m，4H)，3.42-3.37(m，4H)，3.17(m，4H)，2.45-2.43(m，2H)，2.22(m，1H)，2.15(s，3H)。MS(ESI)m/z 519(M++H)。
Example 50: synthesis of Compound 385
(formula 1-2: 4- ((4- (trifluoromethyl) phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (4- (trifluoromethyl) aniline; 0.30g, 1.84mmol) and potassium carbonate (0.76g, 5.53mmol) were dissolved in dimethylformamide (10mL), followed by addition of methyl 4- (bromomethyl) benzoate (0.84g, 3.67 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with water and saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formula 1-2 (0.27g, 47%).
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((4- (trifluoromethyl) phenylamino) methyl) benzoate; 0.27g, 0.87mmol) and 4-nitrophenyl chloroformate (0.35g, 1.74mmol) were dissolved in acetonitrile (10mL), followed by addition of potassium carbonate (0.36g, 2.61 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with water and saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formulae 1-3 (0.40g, 97%).
(formula 1-4: 4- ((N- (4- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.39g, 0.84mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide, followed by addition of potassium carbonate (0.25g, 1.81mmol) and morpholine (10 mL). The mixture was reacted at 60 ℃ for 2 days and diluted with saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compounds of formulae 1-4 (0.20g, 60%).
(Compound 385, N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.20g, 0.48mmol, of the compound of formula 1-4 was dissolved in methanol, and hydroxylamine (50% by weight in water; 0.29mL) and potassium hydroxide (0.13g, 2.39mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give a white solid, and the resulting solid was filtered and dried to give the desired compound 385(0.10g, 50%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(brs，1H)，8.99(brs，1H)，7.67-7.62(m，4H)，7.38(d，2H，J＝8.2Hz)，7.29(d，2H，J＝8.5Hz)，4.94(s，2H)，3.47-3.44(m，4H)，3.21-3.18(m，4H)。MS(ESI)m/z 424(M++H)。
Example 51: synthesis of Compound 386
(formula 1-2: 4- ((3- (trifluoromethyl) phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (3- (trifluoromethyl) aniline; 1.00g, 6.21mmol) and methyl 4-formylbenzoate (1.02g, 6.21mmol) were dissolved in methanol (10ml), and sodium cyanoborohydride (NaCNBH) was added3) (0.41g, 6.21mmol) and acetic acid (0.71mL, 2.00 mmol). The mixture was reacted at room temperature for 2 days, and then methanol was removed under reduced pressure. The reaction was carried out by adding saturated sodium bicarbonateThe solution was complete. The reaction mixture was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formula 1-2 (1.43g, 74.5%).
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((3- (trifluoromethyl) phenylamino) methyl) benzoate; 1.04g, 3.36mmol) and 4-nitrophenyl chloroformate (1.36g, 6.73mmol) were dissolved in acetonitrile (30mL), followed by addition of potassium carbonate (1.39g, 10.1 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with water and a saturated aqueous sodium chloride solution and water, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formula 1-3 (1.51g, 95%).
(formula 1-4: 4- ((2, 6-dimethyl-N- (3- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.38g, 0.80mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (10ml), followed by addition of potassium carbonate (0.33g, 2.38mmol) and 2, 6-dimethylmorpholine (0.09, 0.80 mmol). The mixture was reacted at 60 ℃ for 2 days, then diluted with ethyl acetate and washed with a saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compound of formulae 1-4 (0.21g, 59%).
(Compound 386, N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((2, 6-dimethyl-N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.21g, 0.47mmol, a compound of formulae 1-4 was dissolved in methanol (50mL), and then aqueous hydroxylamine (50 wt%, 0.29mL) and potassium hydroxide (0.13g, 2.35mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give a white solid, and the resulting solid was filtered and dried to give the desired compound 386(0.10g, 48%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.65(d，2H，J＝8.3Hz)，7.54-7.50(m，1H)，7.43-7.36(m，5H)，3.55(d，2H，J＝12.4Hz)，2.36-2.31(m，2H)，0.94(s，3H)，0.93(s，3H)。MS(ESI)m/z 452(M++H)。
Example 52: synthesis of Compound 389
(formula 1-2: 4- ((4-chloro-3- (trifluoromethyl) phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (4-chloro-3- (trifluoromethyl) aniline; 5g, 25.6mmol) and methyl 4-formylbenzoate (4.19g, 25.6mmol) were dissolved in methanol (100mL) and stirred for 1 hour. Then, acetic acid (1.58mL, 25.6mmol) and sodium cyanoborohydride (1.61g, 25.6mmol) were added and stirred for 1 day. Methanol was partially removed by air drying to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound of formula 1-2 (3.4g, 39%) as a white solid.
(formula 1-3: 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (4- ((4-chloro-3- (trifluoromethyl) phenylamino) methyl) benzoate; 2g, 5.82mmol) and 4-nitrophenyl chloroformate (1.17g, 5.82mmol) were dissolved in methylene chloride (50mL) and stirred at room temperature for 2 days to precipitate a solid, and the resulting solid was filtered and dried to give the desired compound of formula 1-3 (3.14g, 106%) as a yellow solid.
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 0.983mmol) and piperidine (0.097mL, 0.983mmol) of the compound of formula 1-3 were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.425g, 95%) as a colorless oil.
(Compound 389: N- (4-chloro-3- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) piperidine-1-carboxamido) methyl) benzoate, 0.263g, 0.578mmol) of the compound of the formula 1-4 was dissolved in methanol (20mL), hydroxylamine hydrochloride (0.201g, 2.89mmol) and potassium hydroxide (0.324g, 5.78mmol) were added and stirring was carried out. Then, hydroxylamine (50 wt% aqueous solution; 1.49mL, 11.56mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 389(0.18g, 68%) as an apricot yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.1Hz)，7.58(d，1H，J＝8.7Hz)，7.43(d，1H，J＝2.6Hz)，7.35(dd，1H，J＝8.8，2.6Hz)，7.28(d，2H，J＝8.1Hz)，4.87(s，2H)，3.71-3.68(m，2H)，2.69-2.63(m，2H)，1.50-1.44(m，2H)，0.87-0.81(m，4H)；MS(ESI)m/z 456.1(M++H)。
Example 53: synthesis of Compound 390
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-methylpiperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 0.983mmol) and 4-methylpiperidine (0.116mL, 0.983mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.369g, 80%) as a white oil.
(Compound 390: N- (4-chloro-3- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperidine-1-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-methylpiperidine-1-carboxamido) methyl) benzoate, 0.239g, 0.51mmol) of the compound of the formula 1-4 was dissolved in methanol (20mL), hydroxylamine hydrochloride (0.177g, 2.55mmol) and potassium hydroxide (0.286g, 5.09mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 1.31mL, 10.19mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 390(0.175g, 73%) as an apricot yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.2Hz)，7.58(d，1H，J＝8.8Hz)，7.44(d，1H，J＝2.6Hz)，7.34(dd，1H，J＝8.8，2.6Hz)，7.29(d，2H，J＝8.2Hz)，4.88(s，2H)，3.18-3.15(m，4H)，1.46-1.45(m，3H)，1.33-1.32(m，5H)；MS(ESI)m/z 470.1(M++H)。
Example 54: synthesis of Compound 391
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 0.983mmol) and morpholine (0.086mL, 0.983mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.229g, 51%) as a white oil.
(Compound 391: N- (4-chloro-3- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.138g, 0.302mmol) of the compound of formula 1-4 was dissolved in methanol (20mL), hydroxylamine hydrochloride (0.105g, 1.51mmol) and potassium hydroxide (0.169g, 3.02mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.778mL, 6.04mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium hydrogencarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 391(0.089g, 64%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(s，1H)，8.99(s，1H)，7.65(d，2H，J＝8.3Hz)，7.60(d，1H，J＝8.7Hz)，7.54(d，1H，J＝2.6Hz)，7.43-7.40(m，1H)，7.36(d，2H，J＝8.2Hz)，4.94(s，2H)，3.44-3.42(m，4H)，3.18-3.16(m，4H)；MS(ESI)m/z 458.1(M++H)。
Example 55: synthesis of Compound 392
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 0.983mmol) and 2, 6-dimethylmorpholine (0.12mL, 0.983mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.2g, 42%) as a white oil.
(Compound 392: N- (4-chloro-3- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethylmorpholine-4-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoate, 0.121g, 0.250mmol) of the compound of formulae 1-4 was dissolved in methanol (20mL), hydroxylamine hydrochloride (0.087g, 1.25mmol) and potassium hydroxide (0.14g, 2.49mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.643mL, 4.99mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 392(0.056g, 46%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.3Hz)，7.59(d，1H，J＝8.8Hz)，7.49(d，1H，J＝2.6Hz)，7.39-7.36(m 1H)，7.30(d，2H，J＝8.2Hz)，4.89(s，2H)，3.58-3.55(m，2H)，2.40-2.32(m，4H)，0.96(d，6H，J＝6.2Hz)；MS(ESI)m/z 486.1(M++H)。
Example 56: synthesis of Compound 393
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 0.983mmol) and 1-methylpiperazine (0.109mL, 0.983mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1 to 4 (0.420g, 91%) as a yellow oil.
(Compound 393: N- (4-chloro-3- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperazine-1-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoate, 0.251g, 0.534mmol) of the compound of the formula 1 to 4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.185g, 2.67mmol) and potassium hydroxide (0.299g, 5.34mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 1.38mL, 10.7mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 393(0.19g, 75%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(s，1H)，8.99(s，1H)，7.65(d，2H，J＝8.1Hz)，7.60(d，1H，J＝8.7Hz)，7.49(d，1H，J＝2.4Hz)，7.40-7.35(m，3H)，5.75(s，1H)，4.92(s，2H)，3.17(m，4H)，2.14(m，4H)，2.10(s，3H)；MS(ESI)m/z 471.1(M++H)。
Example 57: synthesis of Compound 394
(formula 1-4: 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-ethylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((4-chloro-3- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.5g, 0.983mmol) and 1-ethylpiperazine (0.125mL, 0.983mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1 to 4 (0.411g, 86%) as a yellow oil.
(Compound 394: N- (4-chloro-3- (trifluoromethyl) phenyl) -4-ethyl-N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((N- (4-chloro-3- (trifluoromethyl) phenyl) -4-ethylpiperazine-1-carboxamido) methyl) benzoate, 0.261g, 0.539mmol) of the compound of the formula 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.187g, 2.69mmol) and potassium hydroxide (0.303g, 5.39mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 1.39mL, 10.79mmol) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 394(0.25g, 95%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.13(s，1H)，9.00(s，1H)，7.65(d，2H，J＝8.2Hz)，7.60(d，1H，J＝8.8Hz)，7.49(d，1H，J＝2.4Hz)，7.40-7.35(m，3H)，5.75(s，2H)，4.92(s，2H)，3.18(m，4H)，2.24(q，2H，J＝7.2Hz)，2.19(m，4H)，0.93(t，3H，J＝7.1Hz)；MS(ESI)m/z485.1(M++H)。
Example 58: synthesis of Compound 395
(formula 2-2: 4- ((N- (3 ', 5' -difluorobiphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 3, 5-difluorophenylboronic acid (0.088g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 2-2 (0.180g, 84%) as a pale brown solid.
(Compound 395: N- (3 ', 5' -difluorobiphenyl-3-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3 ', 5' -difluorobiphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoate, 0.180g, 0.386mmol, a compound of formula 2-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 2.36mL, 38.6mmol) and potassium hydroxide (0.217g, 3.86mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 395(0.159g, 88%) as a light brown solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.2Hz)，7.57(s，1H)，7.46-7.36(m，6H)，7.23(m，1H)，7.15(m，1H)，4.96(s，2H)，3.40-3.37(m，4H)，3.17-3.14(m，4H)。MS(ESI)m/z468(M++H)。
Example 59: synthesis of Compound 396
(formula 2-2: 4- ((N- (3- (6- (dimethylamino) pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 6-dimethylaminopyridin-3-ylboronic acid (0.092g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 2-2 (0.136g, 62%) as a pale brown solid.
(Compound 396: N- (3- (6- (dimethylamino) pyridin-3-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (6- (dimethylamino) pyridin-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.136g, 0.287mmol, a compound of formula 2-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.75mL, 28.7mmol) and potassium hydroxide (0.161g, 2.87mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 396(0.118g, 87%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ8.38(d，1H，J＝2.5Hz)，7.78(m，1H)，7.64(d，2H，J＝8.3Hz)，7.39-7.35(m，3H)，7.32-7.29(m，2H)，7.05(m，1H)，6.69(d，1H，J＝8.9Hz)，4.93(s，2H)，3.41-3.38(m，4H)，3.18-3.15(m，4H)，3.05(s，6H)。MS(ESI)m/z 476(M++H)。
Example 60: synthesis of Compound 397
(formula 2-2: 4- ((N- (3- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.200g, 0.462mmol), pyrimidin-5-ylboronic acid (0.069g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formula 2-2Material (0.120g, 60%) was in the form of a yellow solid.
(Compound 397: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (pyrimidin-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.120g, 0.277mmol, a compound of formula 2-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.70mL, 27.7mmol) and potassium hydroxide (0.156g, 2.78mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and then extracted by ethyl acetate and saturated aqueous ammonium chloride solution. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered, and then the filtrate was concentrated under reduced pressure to obtain the desired compound 397(0.056g, 47%) as a brown solid.
1H NMR(400MHz，DMSO-d6)δ11.21(brs，1H)，9.18(brs，1H)，9.13(s，2H)，8.97(s，1H)，7.66-7.63(m，3H)，7.51-7.41(m，4H)，7.20(m，1H)，4.97(s，2H)，3.41-3.37(m，4H)，3.19-3.16(m，4H)。MS(ESI)m/z 434(M++H)。
Example 61: synthesis of Compound 398
(formula 2-2: 4- ((N- (3 ', 5' -bis (trifluoromethyl) biphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.200g, 0.462mmol), 3, 5-bis (trifluoromethyl) benzeneBoronic acid (0.143g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to obtain the desired compound of formula 2-2 (0.185g, 71%) as a white solid.
(Compound 398: N- (3 ', 5' -bis (trifluoromethyl) biphenyl-3-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3 ', 5' -bis (trifluoromethyl) biphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.185g, 0.327mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 2.00mL, 32.7mmol) and potassium hydroxide (0.183g, 3.27mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 398(0.160g, 86%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.31(s，2H)，8.09(s，1H)，7.74(s，1H)，7.64(d，2H，J＝8.4Hz)，7.57(m，1H)，7.43-7.40(m，3H)，7.22(m，1H)，4.98(s，2H)，3.41-3.38(m，4H)，3.19-3.16(m，4H)。MS(ESI)m/z 568(M++H)。
Example 62: synthesis of Compound 399
(formula 2-2: 4- ((N- (3- (2, 3-dihydrobenzo [ b))][1，4]IIEn-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.200g, 0.462mmol), 2, 3-dihydrobenzo [ b][1，4]IIEn-6-ylboronic acid (0.100g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formula 2-2 (0.182g, 81%) as a yellow liquid.
(Compound 399: N- (3- (2, 3-dihydrobenzo [ b ]][1，4]IIEn-6-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide
A compound of formula 2-2 (4- ((N- (3- (2, 3-dihydrobenzo [ b ]))][1，4]IIEn-6-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.182g, 0.373mmol) was dissolved in methanol (10mL) and hydroxylamine (50.0 wt% aqueous solution; 2.28mL, 37.3mmol) and potassium hydroxide (0.209g, 3.73mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 399(0.165g, 91%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.2Hz)，7.38(d，2H，J＝8.2Hz)，7.34-7.27(m，3H)，7.12-7.04(m，3H)，6.91(d，1H，J＝8.3Hz)，4.92(s，2H)，4.26(s，4H)，3.40-3.37(m，4H)，3.17-3.15(m，4H)。MS(ESI)m/z 490(M++H)。
Example 63: synthesis of Compound 400
(formula 2-2: 4- ((N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-3-yl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 3, 4, 5-trimethoxyphenylboronic acid (0.117g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was addedThe reaction solution was added and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formula 2-2 (0.146g, 61%) as a white solid.
(Compound 400: N- (4- (hydroxycarbamoyl) benzyl) -N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-3-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (3 ', 4 ', 5 ' -trimethoxybiphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.146g, 0.280mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.72mL, 28.1mmol) and potassium hydroxide (0.157g, 2.81mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 400(0.125g, 86%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝8.2Hz)，7.41-7.39(m，3H)，7.37-7.35(m，2H)，7.12(m，1H)，6.81(s，2H)，4.95(s，2H)，3.83(s，6H)，3.67(s，3H)，3.43-3.40(m，4H)，3.19-3.16(m，4H)。MS(ESI)m/z 522(M++H)。
Example 64: synthesis of Compound 401
(formula 2-2: 4- ((N- (2 ', 6' -dimethylbiphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), 2, 6-dimethylphenylboronic acid (0.083g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formula 2-2 (0.205g, 97%) as a white solid.
(Compound 401: N- (2 ', 6' -dimethylbiphenyl-3-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2 ', 6' -dimethylbiphenyl-3-yl) morpholine-4-carboxamido) methyl) benzoate, 0.205g, 0.447mmol, of the compound of formula 2-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 2.74mL, 44.7mmol) and potassium hydroxide (0.251g, 4.47mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 401(0.195g, 95%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.61(d，2H，J＝8.2Hz)，7.38(t，1H，J＝7.8Hz)，7.33(d，2H，J＝8.3Hz)，7.16-7.11(m，2H)，7.06(d，2H，J＝7.4Hz)，6.81(d，1H，J＝7.6Hz)，6.77(m，1H)，4.86(s，2H)，3.40-3.37(m，4H)，3.17-3.14(m，4H)，1.83(s，6H)。MS(ESI)m/z 460(M++H)。
Example 65: synthesis of Compound 402
(formula 2-2: 4- ((N- (3- (furan-3-yl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.200g, 0.462mmol), furan-3-ylboronic acid (0.062g, 0.554mmol) and Pd (dppf) Cl2(0.038g, 0.046mmol) was dissolved in 1, 4-bisAlkane (3mL), then potassium carbonate (0.128g, 0.923mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 100 ℃ for 4 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate and concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to obtain the desired compound of formula 2-2 (0.113g, 58%) as a brown solid.
(Compound 402: N- (3- (furan-3-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (furan-3-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.113g, 0.269mmol) of the compound of formula 2-2 was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.64mL, 26.9mmol) and potassium hydroxide (0.151g, 2.69mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 402(0.060g, 53%) as a brown solid.
1H NMR(400MHz，DMSO-d6)δ8.20(s，1H)，7.73(s，1H)，7.63(d，2H，J＝8.1Hz)，7.43(s，1H)，7.38(d，2H，J＝8.1Hz)，7.30-7.28(m，2H)，7.00(m，1H)，6.95(m，1H)，4.91(s，2H)，3.39-3.36(m，4H)，3.16-3.13(m，4H)。MS(ESI)m/z 422(M++H)。
Example 66: synthesis of Compound 403
(formula 2-2: 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.400g, 0.923mmol), 1, 2, 3, 6-tetrahydropyran-4-boronic acid pinacol ester (0.233g, 1.11mmol) and Pd (dppf) Cl2(0.030g, 0.046mmol) in 1, 4-bisAlkane (4mL), and cesium carbonate (0.897g, 2.77mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 140 ℃ for 15 minutes by using a microwave reactor. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. It was concentrated under reduced pressure and the residue was purified by column chromatography (silica; B)Ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 2-2 (0.132g, 33%) as a brown solid.
(Compound 403: N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.050g, 0.115mmol, of a compound of formula 2-2 was dissolved in methanol (5mL), and hydroxylamine (50.0 wt% aqueous solution; 1.40mL, 22.9mmol) and potassium hydroxide (0.064g, 1.15mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 403(0.030g, 60%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(s，1H)，8.97(s，1H)，7.63(d，2H，J＝8.3Hz)，7.36(d，2H，J＝8.3Hz)，7.27(t，1H，J＝7.9Hz)，7.18(s，1H)，7.14(d，1H，J＝7.6Hz)，7.03(d，1H，J＝7.8Hz)，6.23(m，1H)，4.88(s，2H)，4.19(d，2H，J＝2.6Hz)，3.78(t，2H，J＝5.5Hz)，3.39-3.36(m，4H)，3.14-3.11(m，4H)，2.38-2.36(m，2H)。MS(ESI)m/z 438(M++H)。
Example 67: synthesis of Compound 404
(formula 2-2: 4- ((N- (3- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.400g, 0.923mmol), 1-methyl-1, 2, 3, 6-tetrahydropyridine-4-boronic acid pinacol ester (0.247g, 1.11mmol) and Pd (dppf) Cl2(0.030g, 0.046mmol) in 1, 4-bisAlkane (4mL), and cesium carbonate (0.897g, 2.77mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 140 ℃ for 15 minutes by using a microwave reactor. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. It was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 2-2 (0.193g, 47%) as a brown solid.
(Compound 404: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.050 g, 0.111mmol) was dissolved in methanol (5mL), and hydroxylamine (50.0 wt% aqueous solution; 1.36mL, 22.2mmol) and potassium hydroxide (0.062g, 1.11mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 404(0.015g, 30%) as a brown solid.
MS(ESI)m/z 451(M++H)。
Example 68: synthesis of Compound 405
(formula 2-2: 4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester)
A compound of formula 2-1 (methyl 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.800g, 1.85mmol), N-Boc-1, 2, 3, 6-tetrahydropyridine-4-boronic acid pinacol ester (0.685g, 2.22mmol) and Pd (dppf) Cl2(0.060g, 0.092mmol) was dissolved in 1, 4-bisAlkane (8mL), and cesium carbonate (1.79g, 5.54mmol) dissolved in water (2mL) was added to the reaction solution and stirred at 140 ℃ for 15 minutes by using a microwave reactor. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. It was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 2-2 (0.832g, 84%) as a brown solid.
(Compound 405: 4- (3- (N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamido) phenyl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester)
The compound of formula 2-2 (4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester; 0.050g, 0.093mmol) was dissolved in methanol (5mL), and hydroxylamine (50.0 wt% aqueous solution; 1.14mL, 18.7mmol) and potassium hydroxide (0.052g, 0.933mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until the volume thereof became about 1mL, and then a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 405(0.030g, 60%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.3Hz)，7.35(d，2H，J＝8.2Hz)，7.26(t，1H，J＝7.9Hz)，7.18(s，1H)，7.12(d，1H，J＝8.2Hz)，7.02(d，1H，J＝8.0Hz)，6.17(m，1H)，4.87(s，2H)，3.97(brs，2H)，3.50(t，2H，J＝5.7Hz)，3.38(t，4H，J＝4.6Hz)，3.12(t，4H，J＝4.5Hz)，2.40(brs，2H)，1.41(s，9H)。MS(ESI)m/z 537.
Example 69: synthesis of Compound 413
(formula 1-2: 4- ((2, 4-difluorophenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (2, 4-difluoroaniline; 3.0g, 23.2mmol) and methyl 4-formylbenzoate (3.81g, 23.2mmol) were dissolved in methanol (500mL) and stirred at room temperature for 2 hours. Then, acetic acid (1.33mL, 23.2mmol) and sodium cyanoborohydride (1.46g, 23.2mmol) were added and stirred for 1 day. Methanol was partially removed by air drying to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound of formula 1-2 (2.9g, 45%) as a white solid.
(formula 1-3: 4- (((2, 4-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((2, 4-difluorophenylamino) methyl) benzoate; 2g, 7.21mmol) and 4-nitrophenyl chloroformate (1.45g, 7.21mmol) were dissolved in dichloromethane (50mL) and stirred at room temperature for 3 days. Then, water was added to extract the organic layer. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was dried to give the desired compound of formulae 1-3 (2.5g, 78%) as a yellow oil.
(formula 1-4: 4- ((N- (2, 4-difluorophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2, 4-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50g, 1.13mmol) and morpholine (0.098mL, 1.13mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1-4 (0.44g, 98%) as a colorless oil.
(Compound 413: N- (2, 4-difluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2, 4-difluorophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.10g, 0.256mmol, of the compound of formulae 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.089g, 1.28mmol) and potassium hydroxide (0.144g, 2.56mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.329mL, 5.12mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Then, the residue was dissolved in dichloromethane and hexane was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 413(0.076g, 76%) as a pale yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.3Hz)，7.41(d，2H，J＝8.2Hz)，7.27-7.25(m，1H)，7.04-6.96(m，2H)，4.80(s，2H)，3.46-3.43(m，4H)，3.22-3.19(m，4H)；MS(ESI)m/z 392.1(M++H)。
Example 70: synthesis of Compound 414
Formulas 1 to 4: 4- ((N- (2, 4-difluorophenyl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2, 4-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50g, 1.13mmol) and piperidine (0.112mL, 1.13mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10%) and concentrated to give the desired compound of formulae 1-4 (0.46g, 104.8%) as a colorless oil.
(Compound 414: N- (2, 4-difluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (2, 4-difluorophenyl) piperidine-1-carboxamido) methyl) benzoate, a compound of formula 1-4 (0.15g, 0.386mmol) was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.134g, 1.93mmol) and potassium hydroxide (0.217g, 3.86mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.496mL, 7.72mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Then, the residue was dissolved in dichloromethane and hexane was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 414(0.118g, 79%) as a pale yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.2Hz)，7.41(d，2H，J＝8.1Hz)，7.23-7.20(m，1H)，7.05-6.94(m，2H)，4.77(s，2H)，3.21-3.18(m，4H)，1.51-1.49(m，2H)，1.32-1.31(m，4H)；MS(ESI)m/z 390.1(M++H)。
Example 71: synthesis of Compound 415
Formulas 1 to 4: 4- ((N- (2, 4-difluorophenyl) 2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2, 4-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 1.13mmol) and 2, 6-dimethylmorpholine (0.138mL, 1.13mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1-4 (0.40g, 85%) as a colorless oil.
(Compound 415: N- (2, 4-difluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethylmorpholine-4-carboxamide)
Methyl 4- ((N- (2, 4-difluorophenyl) 2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoate, 0.20g, 0.478mmol, a compound of formulae 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.166g, 2.39mmol) and potassium hydroxide (0.268g, 4.78mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.614mL, 9.56mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Then, the residue was dissolved in dichloromethane and hexane was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 415(0.086g, 43%) as a pale yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.0Hz)，7.41(d，2H，J＝8.0Hz)，7.27-7.26(m，1H)，7.04-6.94(m，2H)，4.79(s，2H)，3.58(d，2H，J＝12.9Hz)，2.38-2.32(m，2H)，1.01(d，6H，J＝6.2Hz)；MS(ESI)m/z 420.1(M++H)。
Example 72: synthesis of Compound 416
Formulas 1 to 4: 4- ((N- (2, 4-difluorophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2, 4-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.50g, 1.13mmol) and 1-methylpiperazine (0.126mL, 1.13mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1-4 (0.46g, 101%) as a yellow oil.
(Compound 416: N- (2, 4-difluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperazine-1-carboxamide)
Methyl 4- ((N- (2, 4-difluorophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoate of the formula 1 to 4 (0.22g, 0.545mmol) was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.189g, 2.73mmol) and potassium hydroxide (0.306g, 5.45mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.701mL, 10.9mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Then, the residue was dissolved in dichloromethane and hexane was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 416(0.154g, 70%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.2Hz)，7.40(d，2H，J＝8.2Hz)，7.26-7.25(m，1H)，7.04-6.96(m，2H)，4.79(s，2H)，3.25-3.23(m，4H)，2.24-2.21(m，7H)；MS(ESI)m/z 405.1(M++H)。
Example 73: synthesis of Compound 418
(formula 2-2: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-1 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.200g, 0.462mmol), benzo [ d ]][1，3]Dioxol-5-ylboronic acid (0.092g, 0.554mmol) and Pd (dppf) Cl2(0.015g, 0.023mmol) was dissolved in 1, 4-bisAlkane (4mL), and cesium carbonate (0.448g, 1.39mmol) dissolved in water (1mL) was added to the reaction solution and stirred at 140 ℃ for 15 minutes by using a microwave reactor. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate. Then, the organic layer was dehydrated with anhydrous magnesium sulfate and filtered. It was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 2-2 (0.146g, 67%) In the form of a beige solid.
(Compound 418: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-2 (0.146g, 0.308mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.88mL, 30.8mmol) and potassium hydroxide (0.173g, 3.08mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, followed by extraction with ethyl acetate and a saturated aqueous sodium chloride solution. Then, the organic layer was washed with water and dehydrated with anhydrous magnesium sulfate. It was filtered and concentrated under reduced pressure to give the desired compound 418(0.121g, 83%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.2Hz)，7.34-7.27(m，5H)，7.21(d，1H，J＝1.7Hz)，7.09(dd，1H，J＝8.1，1.8Hz)，7.05(d，1H，J＝7.7Hz)，6.97(d，1H，J＝8.1Hz)，6.05(s，2H)，4.90(s，2H)，3.39(t，4H，J＝4.5Hz)，3.16(t，4H，J＝4.4Hz).MS(ESI)m/z 476(M++H)。
Example 74: synthesis of Compound 419
(formula 4-2: 4- ((N- (3- (1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester hydrochloride)
General formulaThe compound 4-1 (4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester; 0.510g, 0.952mmol) was dissolved in 1, 4-bis (tert-butyl) acetateAlkane (5mL) followed by hydrogen chloride (4.0M 1, 4-bis)An alkane solution; 4.76mL, 19.0mmol) and stirred at room temperature for 3 hours. After completion of the reaction, the organic layer was concentrated under reduced pressure, and the residue was recrystallized from dichloromethane and hexane to obtain the desired compound of formula 4-2 (0.440g, 98%) as a brown solid.
(formula 4-3: 4- ((N- (3- (1-acetyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride of the formula 4-2 (0.140g, 0.297mmol) was suspended in dichloromethane (5mL), then N, N-diisopropylethylamine (0.158mL, 0.890mmol) and acetic anhydride (0.036g, 0.356mmol) were successively added, followed by stirring at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formula 4-3 (0.103g, 73%) as a white solid.
(Compound 419: N- (3- (1-acetyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1-acetyl-1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-3 (0.103g, 0.216mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.32mL, 21.6mmol) and potassium hydroxide (0.121g, 2.16mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was washed with water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain the desired compound 419(0.043g, 42%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(s，1H)，8.98(s，1H)，7.63(d，2H，J＝8.2Hz)，7.36(d，2H，J＝8.2Hz)，7.27(t，1H，J＝7.9Hz)，7.18-7.12(m，2H)，7.03(d，1H，J＝8.4Hz)，6.16(m，1H)，4.87(s，2H)，4.12-4.06(m，2H)，3.63-3.58(m，2H)，3.39(t，4H，J＝4.5Hz)，3.12(t，4H，J＝4.4Hz)，2.38(brs，2H)，2.07(d，3H，J＝9.0Hz).MS(ESI)m/z 479(M++H)。
Example 75: synthesis of Compound 420
(formula 4-3: 4- ((N- (3- (1- (methylsulfonyl) -1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride of the formula 4-2 (0.140g, 0.297mmol) was suspended in dichloromethane (5mL), then N, N-diisopropylethylamine (0.158mL, 0.890mmol) and methanesulfonyl chloride (0.040g, 0.356mmol) were successively added, followed by stirring at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70%) and concentrated to give the desired compound of formula 4-3 (0.114g, 75%) as a white solid.
(Compound 420: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1- (methylsulfonyl) -1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1- (methylsulfonyl) -1, 2, 3, 6-tetrahydropyridin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-3 (0.114g, 0.222mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 1.36mL, 22.2mmol) and potassium hydroxide (0.125g, 2.22mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 2mL, and the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was washed with water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to give the desired compound 420(0.038g, 33%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.0Hz)，7.34(d，2H，J＝7.9Hz)，7.27(t，1H，J＝7.8Hz)，7.20(s，1H)，7.14(d，1H，J＝8.3Hz)，7.04(d，1H，J＝7.0Hz)，6.19(s，1H)，4.85(s，2H)，3.83(d，2H，J＝2.2Hz)，3.39-3.35(m，6H)，3.14-3.11(m，4H)，2.92(s，3H)，2.54(m，2H)。MS(ESI)m/z 515(M++H)。
Example 76: synthesis of Compound 438
(formula 1-2: 4- ((2-fluoro-4-methylphenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (2-fluoro-4-methylaniline; 3.0g, 23.9mmol) and methyl 4-formylbenzoate (3.94g, 23.9mmol) were dissolved in methanol (500mL) and stirred at room temperature for 2 hours. Then, acetic acid (1.44mL, 23.9mmol) and sodium cyanoborohydride (1.51g, 23.9mmol) were added and stirred for 1 day. Methanol was partially removed by air drying to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound of formula 1-2 (4.2g, 64%) as a white solid.
(formula 1-3: 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (4- ((2-fluoro-4-methylphenylamino) methyl) benzoic acid methyl ester; 2g, 7.32mmol) and 4-nitrophenyl chloroformate (1.48g, 7.32mmol) were dissolved in methylene chloride (50mL) and stirred at room temperature for 3 days, and then the organic layer was extracted with water. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was dried to give the desired compound of formulae 1-3 (2.5g, 78%) as a yellow solid.
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) piperidine-1-formamido) methyl) benzoate)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50g, 1.14mmol) and piperidine (0.113mL, 1.14mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10%) and concentrated to give the desired compound of formulae 1-4 (0.56g, 127%) as a white oil.
(Compound 438: N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) piperidine-1-carboxamido) methyl) benzoate, 0.281g, 0.731mmol) of the compound of formula 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.254g, 3.66mmol) and potassium hydroxide (0.41g, 7.31mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.94mL, 14.6mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 438(0.23g, 82%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.61(d，2H，J＝8.2Hz)，7.34(d，2H，J＝8.2Hz)，7.13-7.08(m，1H)，7.03(d，1H，J＝11.6Hz)，6.94(d，1H，J＝8.2Hz)，4.68(s，2H)，3.09-3.06(m，4H)，2.24(s，3H)，1.39(m，2H)，1.19(m，4H)；MS(ESI)m/z 386.1(M++H)。
Example 77: synthesis of Compound 439
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50g, 1.14mmol) and morpholine (0.099mL, 1.14mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.50g, 114%) as a white oil.
(Compound 439: N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) morpholine-4-carboxamido) methyl) benzoate, 0.213g, 0.551mmol, a compound of formula 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.191g, 2.76mmol) and potassium hydroxide (0.309g, 5.51mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.709mL, 11.0mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 439(0.175g, 82%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(s，1H)，9.01(s，1H)，7.62(d，2H，J＝8.2Hz)，7.33(d，2H，J＝8.2Hz)，7.17-7.13(m，1H)，7.05(d，1H，J＝11.6Hz)，6.96(d，1H，J＝8.2Hz)，4.72(s，2H)，3.33-3.29(m，4H)，3.09-3.06(m，4H)，3.42(s，3H)；MS(ESI)m/z 388.2(M++H)。
Example 78: synthesis of Compound 440
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) 2, 6-dimethylmorpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50 g, 1.14mmol) and 2, 6-dimethylmorpholine (0.14mL, 1.14mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.27g, 57%) as a white oil.
(Compound 440: N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethylmorpholine-4-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoate, a compound of formula 1-4 (0.104g, 0.251mmol) was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.087g, 1.26mmol) and potassium hydroxide (0.141g, 2.51mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.323mL, 5.02mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 440(0.086g, 83%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.60(d，2H，J＝7.8Hz)，7.27(d，2H，J＝6.7Hz)，7.14-7.12(m，1H)，7.03(d，1H，J＝11.8Hz)，6.94(d，1H，J＝8.0Hz)，4.68(s，2H)，3.49(d，2H，J＝12.7Hz)，3.19-3.17(m，2H)，2.27-2.21(m，2H)，0.90(d，6H，J＝6.1Hz)；MS(ESI)m/z 416.2(M++H)。
Example 79: synthesis of Compound 441
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) 4-methylpiperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.50 g, 1.14mmol) and 1-methylpiperazine (0.127mL, 1.14mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 5%) and concentrated to give the desired compound of formulae 1 to 4 (0.43g, 94%) as a yellow oil.
(Compound 441: N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperazine-1-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) 4-methylpiperazine-1-carboxamido) methyl) benzoate, 0.095g, 0.238mmol, a compound of formula 1-4 was dissolved in methanol (20mL), and hydroxylamine hydrochloride (0.083g, 1.19mmol) and potassium hydroxide (0.133g, 2.38mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.306mL, 4.76mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 441(0.008g, 8%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(s，1H)，9.01(s，1H)，7.61(d，2H，J＝8.3Hz)，7.32(d，2H，J＝8.2Hz)，7.14-7.10(m，1H)，7.05-7.02(m，1H)，6.95(d，1H，J＝8.2Hz)，4.70(s，2H)，3.09-3.07(m，4H)，2.24(s，3H)，2.05-2.02(m，4H)；MS(ESI)m/z 401.2(M++H)。
Example 80: synthesis of Compound 450
(formula 7-6: 4- ((4-acetyl-N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.40g, 0.74mmol) was dissolved in dimethylformamide (10mL), then 1- (1, 4-diazepan-1-yl) ethanone (0.102mL, 1.08mmol) was added and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formula 7-6 (0.378g, 94%) as a white oil.
(Compound 450: 4-acetyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamide)
Methyl 4- ((4-acetyl-N- (pyridin-2-yl) -1, 4-diazepan-1-carboxamido) methyl) benzoate, a compound of formula 7-6 (0.20g, 0.487mmol) was dissolved in methanol (20mL), and hydroxylamine (0.169g, 2.44mmol) and potassium hydroxide (0.273g, 4.87mmol) were added. Then, hydroxylamine (50 wt% aqueous solution; 0.626mL, 9.75mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 450(0.07g, 35%) as a yellow oil.
1H NMR(400MHz，MeOD-d3)δ8.33-8.30(m，1H)，7.74-7.63(m，3H)，7.51-7.46(m，2H)，7.06-6.99(m，2H)，4.89(s，2H)，3.58-3.27(m，8H，1.99-1.96(m，3H)，1.76-1.57(m，2H)；MS(ESI)m/z 412.2(M++H)。
Example 81: synthesis of Compound 451
(formula 7-6: 4- ((4- (cyclopropanecarbonyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 7-5 (4- ((((4-nitrophenoxy) carbonyl) (pyridin-2-yl) amino) methyl) benzoate; 0.40g, 0.982mmol) was dissolved in dimethylformamide (10mL), then cyclopropyl (piperazin-1-yl) methanone (0.167mL, 1.17mmol) was added and the mixture was heated and stirred at 60 ℃ for 2 days. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formula 7-6 (0.4g, 96%) as a white oil.
(Compound 451: 4- (Cyclopropanecarbonyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (pyridin-2-yl) piperazine-1-carboxamide)
Methyl 4- ((4- (cyclopropanecarbonyl) -N- (pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoate, 0.20g, 0.473mmol of the compound of formula 7-6 was dissolved in methanol (20mL), and hydroxylamine (0.164g, 2.38mmol) and potassium hydroxide (0.265g, 4.73mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.61mL, 9.47mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 451(0.05g, 25%) as a yellow oil.
1H NMR(400MHz，MeOD-d3)δ8.33(d，1H，J＝3.7Hz)，7.75-7.71(m，1H)，7.67(d，2H，J＝8.2Hz)，7.49(d，2H，J＝8.2Hz)，7.10(d，1H，J＝8.3Hz)，7.06-7.03(m，1H)，5.05(s，2H)，3.67(m，4H)，3.31(m，4H)，1.89-1.86(m，1H)，0.89-0.88(m，4H)；MS(ESI)m/z 424.2(M++H)。
Example 82: synthesis of Compound 453
(formula 1-4: 4- ((4-Ethyl-N- (2-fluoro-4-methylphenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.20g, 0.456mmol) and 1-ethylpiperazine (0.116mL, 0.912mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1-4 (0.14g, 74%) as a yellow oil.
(Compound 453: 4-Ethyl-N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((4-ethyl-N- (2-fluoro-4-methylphenyl) piperazine-1-carboxamido) methyl) benzoate, 0.14g, 0.339mmol, a compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.118g, 1.69mmol) and potassium hydroxide (0.19g, 3.39mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.435mL, 6.77mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium hydrogencarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 453(0.12g, 86%) as a yellow oil.
1H NMR(400MHz，MeOD-d3)δ7.64(d，2H，J＝8.2Hz)，7.33(d，2H，J＝8.2Hz)，7.06-6.93(m，3H)，4.77(s，2H)，3.26-3.24(m，4H)，2.34(q，2H，J＝7.2Hz)，2.29(s，3H)，2.26-2.23(m，4H)，1.03(t，3H，J＝7.2Hz)；MS(ESI)m/z 415.2(M++H)。
Example 83: synthesis of Compound 454
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) -4- (2-methoxyphenyl) piperazine-1-formamido) methyl) benzoate)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3, 0.20g, 0.456mmol and 4- (2-methoxyphenyl) piperazine (0.175mL, 0.912mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.15g, 67%) as a yellow oil.
(Compound 454: N- (2-fluoro-4-methylphenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4- (2-methoxyphenyl) piperazine-1-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) -4- (2-methoxyphenyl) piperazine-1-carboxamido) methyl) benzoate, 0.15g, 0.305mmol, a compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.106g, 1.53mmol) and potassium hydroxide (0.171g, 3.05mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.392mL, 6.10mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 454(0.06g, 43%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.67(d，2H，J＝8.0Hz)，7.44(d，2H，J＝8.0Hz)，7.14-7.10(m，1H)，7.03-6.84(m，6H)，4.83(s，2H)，3.82(s，3H)，3.38(m，4H)，2.80(m，4H)，2.33(s，3H)；MS(ESI)m/z 493.2(M++H)。
Example 84: synthesis of Compound 455
(formula 1-4: 4- ((N- (2-fluoro-4-methylphenyl) -4- (4-fluorophenyl) piperazine-1-formamido) methyl) benzoate)
Methyl 4- (((2-fluoro-4-methylphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3, 0.20g, 0.456mmol and 1- (4-fluorophenyl) piperazine (0.164mL, 0.912mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formulae 1-4 (0.12g, 55%) as a yellow oil.
(Compound 455: N- (2-fluoro-4-methylphenyl) -4- (4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((N- (2-fluoro-4-methylphenyl) -4- (4-fluorophenyl) piperazine-1-carboxamido) methyl) benzoate, 0.12g, 0.25mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.087g, 1.25mmol) and potassium hydroxide (0.14g, 2.50mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.322mL, 5.01mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 455(0.051g, 42%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.65(d，2H，J＝8.1Hz)，7.38(d，2H，J＝8.1Hz)，7.10-7.06(m，1H)，6.99-6.86(m，6H)，4.80(s，2H)，3.38-3.35(m，4H)，2.87-2.84(m，4H)，2.30(s，3H)；MS(ESI)m/z 481.2(M++H)。
Example 85: synthesis of Compound 456
(formula 1-2: 4- ((3-chloro-4-fluorophenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (3-chloro-4-fluoroaniline; 2.0g, 13.7mmol) and methyl 4-formylbenzoate (2.26g, 13.7mmol) were dissolved in methanol (500mL) and stirred at room temperature for 3 hours. Then, acetic acid (0.786mL, 13.7mmol) and sodium cyanoborohydride (0.86g, 13.7mmol) were added and stirred for 1 day. Methanol was partially removed by air drying to precipitate a solid, and the resulting solid was filtered and dried to give the desired compound of formula 1-2 (2.9g, 72%) as a gray solid.
(formula 1-3: 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((3-chloro-4-fluorophenylamino) methyl) benzoate; 2.5g, 8.51mmol) and 4-nitrophenylchloroformate (2.06g, 10.2mmol) were dissolved in methylene chloride (50mL) and stirred at room temperature for 3 days, and then the organic layer was extracted with water. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was dried to give the desired compound of formulae 1-3 (2.5g, 64%) as a purple oil.
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.20g, 0.436mmol) and piperidine (0.043mL, 0.436mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.15g, 85%) as a white solid.
(Compound 456: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) piperidine-1-carboxamido) methyl) benzoate, 0.10g, 0.247mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.086g, 1.23mmol) and potassium hydroxide (0.139g, 2.47mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.318mL, 4.94mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 456(0.066g, 66%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.2Hz)，7.35-7.29(m，4H)，7.10-7.06(m，1H)，4.82(s，2H)，3.14-3.12(m，4H)，1.44-1.43(m，2H)，1.30(m，4H)；MS(ESI)m/z 406.1(M++H)。
Example 86: synthesis of Compound 457
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) 4-methylpiperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.20g, 0.436mmol) and 4-methylpiperidine (0.051mL, 0.436mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.17g, 93%) as a yellow oil.
(Compound 457: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperidine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) 4-methylpiperidine-1-carboxamido) methyl) benzoate of the formula 1-4; 0.10g, 0.239mmol) was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.083g, 1.19mmol) and potassium hydroxide (0.134g, 2.39mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.307mL, 4.78mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 457(0.04g, 40%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.2Hz)，7.35-7.31(m，4H)，7.06-7.04(m，1H)，4.82(s，2H)，3.69-3.66(m，2H)，2.59(m，2H)，1.46-1.44(m，3H)，0.82-0.81(m，5H)；MS(ESI)m/z 420.2(M++H)。
Example 87: synthesis of Compound 458
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3 (0.20g, 0.436mmol) and morpholine (0.038mL, 0.436mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.022g, 12%) as a colorless oil.
(Compound 458: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) morpholine-4-carboxamido) methyl) benzoate of formula 1-4; 0.050g, 0.123mmol) was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.043g, 0.614mmol) and potassium hydroxide (0.069g, 1.23mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.158mL, 2.46mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 458(0.017g, 34%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.2Hz)，7.44-7.42(m，1H)，7.33-7.29(m，3H)，7.15-7.11(m，1H)，4.84(s，2H)，3.41-3.40(m，4H)，3.14-3.12(m，4H)；MS(ESI)m/z408.1(M++H)。
Example 88: synthesis of Compound 459
(formula 1-4: 4- ((4-methyl-N- (3- (trifluoromethyl) phenyl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.38g, 0.80mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (10mL), followed by addition of potassium carbonate (0.33g, 2.38mmol) and 4-methylpiperidine (0.10mL, 0.80 mmol). The mixture was reacted at 60 ℃ for 2 days, then diluted with ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compound of formulae 1-4 (0.26g, 75%).
(Compound 459: N- (4- (hydroxycarbamoyl) benzyl) -4-methyl-N- (3- (trifluoromethyl) phenyl) piperidine-1-carboxamide)
Methyl 4- ((4-methyl-N- (3- (trifluoromethyl) phenyl) piperidine-1-carboxamido) methyl) benzoate, 0.26g, 0.60mmol, a compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.37mL) and potassium hydroxide (0.17g, 2.98mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give the product as a white solid, and the resulting solid was filtered and dried to give the desired compound 459(0.07g, 26%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.65(d，2H，J＝8.3Hz)，7.54-7.50(m，1H)，7.43-7.36(m，5H)，4.92(s，2H)，3.41-3.36(m，4H)，2.36-2.26(m，3H)，0.94(s，3H)。
Example 89: synthesis of Compound 460
(formula 1-4: 4- ((N- (3- (trifluoromethyl) phenyl) piperidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.44g, 0.94mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (10mL), followed by addition of potassium carbonate (0.39g, 2.81mmol) and piperidine (0.09mL, 0.94 mmol). The mixture was reacted at 60 ℃ for 1 day, then diluted with ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate hexanes ═ 20%) to give the desired compounds of formulae 1-4 (0.16g, 41%).
(Compound 460: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperidine-1-carboxamide)
Methyl 4- ((N- (3- (trifluoromethyl) phenyl) piperidine-1-carboxamido) methyl) benzoate, 0.27g, 0.65mmol, a compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.40mL) and potassium hydroxide (0.18g, 3.27mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give the solid product, and the resulting solid was filtered and dried to give the desired compound 460(0.07g, 24%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝8.2Hz)，7.53-7.49(m，1H)，7.38-7.32(m，5H)，7.15-7.11(m，1H)，4.91(s，2H)，3.17-3.14(m，4H)，1.45-1.46(m，2H)，1.30(brs，4H)；MS(ESI)m/z 422.1(M++H)。
Example 90: synthesis of Compound 461
(formula 1-4: 4- ((4-Ethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.346g, 0.73mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (10mL), followed by the addition of potassium carbonate (0.30g, 2.19mmol) and 1-ethylpiperazine (0.09mL, 0.73 mmol). The mixture was reacted at 60 ℃ for 1 day, then diluted with ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate hexanes ═ 50%) to give the desired compounds of formulae 1-4 (0.15g, 46%).
(Compound 461: 4-Ethyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4-ethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.15g, 0.33mmol, of the compound of formula 1-4 was dissolved in methanol (10mL), and hydroxylamine (50 wt% aqueous solution; 0.20mL) and potassium hydroxide (0.09g, 1.67mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was stirred in diethyl ether to give a solid product, and the resulting solid was filtered and dried to give the desired compound 461(0.09g, 61%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，7.65(d，2H，J＝8.2Hz)，7.51(t，1H，J＝7.9Hz)，7.41-7.36(m，5H)，4.92(s，2H)，3.17-3.14(m，4H)，2.25，2.22(ABq，2H，J＝12.4，7.2Hz)，2.18-2.15(m，4H)，0.92(t，3H，J＝7.2Hz)；MS(ESI)m/z 451.1(M++H)。
Example 91: synthesis of Compound 462
(formula 5-2: 4- ((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenylamino) methyl) benzoic acid methyl ester)
A compound of formula 5-1 (methyl 4- ((3-bromophenylamino) methyl) benzoate; 3.00g, 9.37mmol), benzo [ d][1，3]Dioxol-5-ylboronic acid (1.87g, 11.2mmol) and Pd (dbpf) Cl2(0.305g, 0.468mmol) was dissolved in 1, 4-bisAlkane (24mL) and sodium carbonate (1.99g, 18.7mmol) dissolved in water (6mL) was added to the reaction solution and stirred in a microwave reactor at 120 ℃ for 30 minutes. After completion of the reaction, the organic layer was washed with ethyl acetate and saturated aqueous sodium bicarbonateExtracted, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10%) and concentrated to give the desired compound of formula 5-2 (1.52g, 45%) as a brown solid.
(formula 5-3: 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
Methyl 4- ((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenylamino) methyl) benzoate of formula 5-2, 1.52g, 4.21mmol and 4-nitrophenyl chloroformate (0.933g, 4.63mmol) were dissolved in acetonitrile (20mL), followed by addition of potassium carbonate (0.872g, 6.31mmol) and stirring at room temperature for 3 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 5-3 (1.87g, 84%) as a pale yellow solid.
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of piperidine (0.087g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.155g, 96%) as a pale yellow liquid.
(Compound 462: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) piperidine-1-carboxamido) methyl) benzoate, a compound of formula 5-4 (0.155g, 0.328mmol) was dissolved in methanol (10mL), followed by the sequential addition of hydroxylamine (50.0 wt.% aqueous solution; 1.00mL, 16.4mmol) and potassium hydroxide (0.184g, 3.28mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 462(0.153g, 99%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.3Hz)，7.39(d，2H，J＝8.2Hz)，7.36-7.25(m，3H)，7.19(d，1H，J＝1.8Hz)，7.08(dd，1H，J＝8.1，1.8Hz)，7.02-6.95(m，2H)，6.05(s，2H)，4.90(s，2H)，3.17-3.14(m，4H)，1.43(m，2H)，1.28(m，4H)。MS(ESI)m/z 474(M++H)。
Example 92: synthesis of Compound 463
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of 4-hydroxypiperidine (0.104g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formula 5-4 (0.128g, 77%) as a pale yellow liquid.
(Compound 463: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxy-N- (4- (hydroxycarbamoyl) benzyl) piperidine-1-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) benzoate (formula 5-4) (0.128g, 0.262mmol) was dissolved in methanol (10mL), followed by the sequential addition of hydroxylamine (50.0 wt% aqueous solution; 0.801mL, 13.1mmol) and potassium hydroxide (0.147g, 2.62mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 463(0.085g, 66%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ7.64(d，2H，J＝8.3Hz)，7.39(d，2H，J＝8.3Hz)，7.33-7.27(m，3H)，7.20(d，1H，J＝1.8Hz)，7.08(dd，1H，J＝8.1，1.8Hz)，7.02-7.00(m，1H)，6.97(d，1H，J＝8.1Hz)，6.05(s，2H)，4.90(s，2H)，4.65(brs，1H)，3.54-3.51(m，3H)，2.87-2.81(m，2H)，1.56-1.53(m，2H)，1.14-1.09(m，2H)。MS(ESI)m/z 490(M++H)。
Example 93: synthesis of Compound 464
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of 2, 6-dimethylmorpholine (0.118g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 25%) and concentrated to give the desired compound of formula 5-4 (0.116g, 68%) as a colorless liquid.
(Compound 464: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethylmorpholine-4-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoate, a compound of formula 5-4 (0.116g, 0.231mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.706mL, 11.5mmol) and potassium hydroxide (0.130g, 2.31mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 464(0.085g, 73%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.3Hz)，7.40(d，2H，J＝8.2Hz)，7.37-7.27(m，3H)，7.21(d，1H，J＝1.8Hz)，7.09(dd，1H，J＝8.1，1.8Hz)，7.04(m，1H)，6.98(d，1H，J＝8.1Hz)，6.05(s，2H)，4.90(s，2H)，3.61-3.58(m，2H)，3.29-3.27(m，2H)，2.30-2.27(m，2H)，0.92(s，3H)，0.90(s，3H)。MS(ESI)m/z 504(M++H)。
Example 94: synthesis of Compound 465
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxy-4-phenylpiperidine-1-carboxamido) methyl) benzoic acid methyl ester)
That is, methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (formula 5-3; 0.180g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of 4-phenylpiperidin-4-ol (0.182g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.137g, 71%) as a pale yellow liquid.
(Compound 465: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxy-N- (4- (hydroxycarbamoyl) benzyl) -4-phenylpiperidine-1-carboxamide)
A compound of formula 5-4 (methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxy-4-phenylpiperidine-1-carboxamido) methyl) benzoate; 0.137g, 0.243mmol) was dissolved in methanol (10mL), followed by successively adding hydroxylamine (50.0 wt% aqueous solution; 0.742mL, 12.1mmol) and potassium hydroxide (0.136g, 2.43mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 465(0.118g, 86%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.66(d，2H，J＝8.2Hz)，7.41(d，2H，J＝7.9Hz)，7.38-7.33(m，2H)，7.29-7.20(m，6H)，7.17-7.06(m，3H)，6.99(d，1H，J＝8.2Hz)，6.06(s，2H)，4.98(s，1H)，4.92(s，2H)，3.69(d，2H，J＝11.8Hz)，3.04(t，2H，J＝12.2Hz)，1.63-1.58(m，2H)，1.40-1.36(m，2H)。MS(ESI)m/z 566(M++H)。
Example 95: synthesis of Compound 466
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 5-3, methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.180g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of 1-methylpiperazine (0.103g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 5-4 (0.099g, 59%) as a yellow liquid.
(Compound 466: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperazine-1-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoate of the formula 5-4 (0.099g, 0.203mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0% by weight aqueous solution; 0.621mL, 10.2mmol) and potassium hydroxide (0.114g, 2.03mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous sodium bicarbonate solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 466(0.036g, 36%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.1Hz)，7.35(d，2H，J＝8.2Hz)，7.32-7.26(m，3H)，7.21(d，1H，J＝1.8Hz)，7.09(dd，1H，J＝8.2，1.7Hz)，7.02(m，1H)，6.98(d，1H，J＝8.1Hz)，6.05(s，2H)，4.90(s，2H)，3.17(m，4H)，2.10(m，4H)，2.06(s，3H)。MS(ESI)m/z489(M++H)。
Example 96: synthesis of Compound 467
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4- (2-hydroxyethyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 5-3, methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.180g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of 2- (piperazin-1-yl) ethanol (0.134g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 5-4 (0.101g, 57%) as a yellow liquid.
(Compound 467: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4- (2-hydroxyethyl) piperazine-1-carboxamide)
A compound of formula 5-4 (methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4- (2-hydroxyethyl) piperazine-1-carboxamido) methyl) benzoate; 0.101g, 0.195mmol) was dissolved in methanol (10mL), followed by the sequential addition of hydroxylamine (50.0 wt% aqueous solution; 0.597mL, 9.76mmol) and potassium hydroxide (0.110g, 1.95mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL and extracted with ethyl acetate and a saturated aqueous sodium bicarbonate solution. The organic was washed with water and dehydrated over anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to obtain the desired compound 467(0.054g, 53%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.12(s，1H)，9.00(s，1H)，7.64(d，2H，J＝8.2Hz)，7.38(d，2H，J＝8.2Hz)，7.34-7.26(m，3H)，7.21(d，1H，J＝1.7Hz)，7.09(dd，1H，J＝8.2，1.8Hz)，7.02(d，1H，J＝7.9Hz)，6.98(d，1H，J＝8.1Hz)，6.05(s，2H)，4.91(s，2H)，4.36(t，1H，J＝5.3Hz)，3.41(q，2H，J＝5.9Hz)，3.16(m，4H)，2.27(t，2H，J＝6.2Hz)，2.22(m，4H)。MS(ESI)m/z 519(M++H)。
Example 97: synthesis of Compound 468
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), then pyrrolidine (0.073g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) were added and stirred at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.148g, 94%) as a colorless liquid.
(Compound 468: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) pyrrolidine-1-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate of formula 5-4 (0.148g, 0.323mmol) was dissolved in methanol (10mL), followed by addition of hydroxylamine (50.0 wt% aqueous solution; 0.987mL, 16.1mmol) and potassium hydroxide (0.181g, 3.23mmol) successively, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to afford the desired compound 468(0.137g, 92%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.1Hz)，7.38(d，2H，J＝8.2Hz)，7.31-7.28(m，3H)，7.18(d，1H，J＝1.7Hz)，7.05(dd，1H，J＝8.1，1.9Hz)，7.00-6.95(m，2H)，6.04(s，2H)，4.91(s，2H)，3.04(m，4H)，1.64(m，4H)。MS(ESI)m/z 460(M++H)。
Example 98: synthesis of Compound 469
(formula 5-4 (S) -4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -2 (hydroxymethyl) pyrrolidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of (S) -pyrrolidin-2-yl-methanol (0.104g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.142g, 85%) as a colorless liquid.
(Compound 469 (S) -N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2- (hydroxymethyl) pyrrolidine-1-carboxamide)
Methyl ((S) -4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -2 (hydroxymethyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.142g, 0.291mmol) of the compound of formula 5-4 was dissolved in methanol (10mL), followed by addition of hydroxylamine (50.0 wt% aqueous solution; 0.889mL, 14.5mmol) and potassium hydroxide (0.163g, 2.91mmol) successively, and stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. Then, the solid product was filtered, washed with water, and then dried in vacuo to give the desired compound 469(0.112g, 79%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.2Hz)，7.39-7.36(m，3H)，7.31-7.25(m，2H)，7.20(d，1H，J＝1.8Hz)，7.11-7.07(m，2H)，6.95(d，1H，J＝8.1Hz)，6.04(s，2H)，5.01(d，1H，J＝16.1Hz)，4.85-4.81(m，2H)，3.93(m，1H)，3.54(m，2H)，3.10(m，1H)，2.55(m，1H)，1.82(m，1H)，1.65(m，2H)，1.56(m，1H)。MS(ESI)m/z 490(M++H)。
Example 99: synthesis of Compound 470
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4- (cyclopropanecarbonyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 5-3, methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.180g, 0.342mmol) was dissolved in dimethylformamide (2mL), followed by addition of cyclopropyl (piperazin-1-yl) methanone (0.158g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) and stirring at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.096g, 52%) as a yellow liquid.
(Compound 470: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4- (cyclopropanecarbonyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
A compound of formula 5-4 (methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4- (cyclopropanecarbonyl) piperazine-1-carboxamido) methyl) benzoate; 0.096g, 0.177mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.542mL, 8.86mmol) and potassium hydroxide (0.100g, 1.77mmol) were successively added, followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous ammonium chloride solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried in vacuo to give the desired compound 470(0.048g, 50%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.1Hz)，7.40-7.38(m，3H)，7.33-7.31(m，2H)，7.23(d，1H，J＝1.6Hz)，7.10(dd，1H，J＝8.1，1.6Hz)，7.06(m，1H)，6.97(d，1H，J＝8.2Hz)，6.05(s，2H)，4.93(s，2H)，3.50(m，4H)，3.21-3.17(m，4H)，1.88(m，1H)，0.68-0.63(m，4H)。MS(ESI)m/z 543(M++H)。
Example 100: synthesis of Compound 471
(formula 5-4: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) azetidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 5-3 (0.180 g, 0.342mmol) was dissolved in dimethylformamide (2mL), and then azetidine hydrochloride (0.096g, 1.03mmol) and potassium carbonate (0.142g, 1.03mmol) were added and stirred at 50 ℃ for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formula 5-4 (0.150g, 99%) as a white solid.
(Compound 471: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) azetidine-1-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) azetidine-1-carboxamido) methyl) benzoate, a compound of formula 5-4 (0.150g, 0.337mmol) was dissolved in methanol (10mL), followed by the sequential addition of hydroxylamine (50.0 wt% aqueous solution; 1.03mL, 16.9mmol) and potassium hydroxide (0.189g, 3.38mmol), followed by stirring at room temperature for 30 minutes. After the completion of the reaction, the reaction solution was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous sodium bicarbonate solution (1-2mL) was added and stirred. The solid product was then filtered, washed with water, and dried under vacuum to give the desired compound 471(0.150g, 99%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.2Hz)，7.40-7.37(m，2H)，7.34-7.30(m，3H)，7.19(d，1H，J＝1.8Hz)，7.08-7.05(m，2H)，6.97(d，1H，J＝8.1Hz)，6.04(s，2H)，4.90(s，2H)，3.52-3.48(m，4H)，1.94-1.92(m，2H)。MS(ESI)m/z 446(M++H)。
Example 101: synthesis of Compound 477
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) -2, 6-dimethylmorpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.20g, 0.436mmol) and 2, 6-dimethylmorpholine (0.053mL, 0.436mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.048g, 25%) as a yellow oil.
(Compound 477: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethylmorpholine-4-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) -2, 6-dimethylmorpholine-4-carboxamido) methyl) benzoate, 0.048g, 0.11mmol, a compound of formulae 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.038g, 0.552mmol) and potassium hydroxide (0.062g, 1.10mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.142mL, 2.21mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 477(0.022g, 46%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.67(d，2H，J＝7.7Hz)，7.40(d，2H，J＝7.6Hz)，7.31-7.30(m，1H)，7.22-7.18(m，1H)，7.11-7.09(m，1H)，3.66-3.62(m，2H)，3.43-3.39(m，2H)，2.40-2.34(m，2H)，1.03(d，6H，J＝6.1Hz)；MS(ESI)m/z 436.1(M++H)。
Example 102: synthesis of Compound 478
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.20g, 0.436mmol) and 1-methylpiperazine (0.049mL, 0.436mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1 to 4 (0.13g, 72%) as a yellow oil.
(Compound 478: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4-methylpiperazine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) -4-methylpiperazine-1-carboxamido) methyl) benzoate of the formula 1 to 4; 0.10g, 0.238mmol) was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.083g, 1.19mmol) and potassium hydroxide (0.134g, 2.38mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.307mL, 4.76mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 478(0.077g, 77%) as a yellow oil.
1H NMR(400MHz，MeOD-d3)δ7.67(d，2H，J＝8.1Hz)，7.38(d，2H，J＝8.1Hz)，7.30-7.28(m，1H)，7.22-7.17(m，1H)，7.10-7.07(m，1H)，4.89(s，2H)，3.29-3.27(m，4H)，2.30-2.27(m，4H)，2.23(s，3H)；MS(ESI)m/z 421.1(M++H)。
Example 103: synthesis of Compound 479
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) -4-ethylpiperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3, 0.20g, 0.436mmol and 1-ethylpiperazine (0.055mL, 0.436mmol) were dissolved in dimethylformamide (10mL), and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 5%) and concentrated to give the desired compound of formulae 1-4 (0.165g, 87%) as a yellow oil.
(Compound 479: N- (3-chloro-4-fluorophenyl) -4-ethyl-N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) -4-ethylpiperazine-1-carboxamido) methyl) benzoate of the formula 1 to 4; 0.10g, 0.23mmol) was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.08g, 1.15mmol) and potassium hydroxide (0.129g, 2.31mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.297mL, 4.61mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 479(0.092g, 92%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.67(d，2H，J＝8.2Hz)，7.39(d，2H，J＝8.2Hz)，7.30-7.28(m，1H)，7.22-7.17(m，1H)，7.10(m，1H)，4.90(s，2H)，3.31-3.28(m，4H)，2.41-2.36(m，2H)，2.33-2.31(m，4H)，1.06(t，3H，J＝7.2Hz)；MS(ESI)m/z 435.1(M++H)。
Example 104: synthesis of Compound 480
(formula 1-4: 4- ((4-benzyl-N- (3-chloro-4-fluorophenyl) piperazine-1-formamido) methyl) benzoate)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.20g, 0.436mmol) and 1-benzylpiperazine (0.075mL, 0.436mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.151g, 70%) as a yellow oil.
(Compound 480: 4-benzyl-N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((4-benzyl-N- (3-chloro-4-fluorophenyl) piperazine-1-carboxamido) methyl) benzoate, 0.10g, 0.202mmol, a compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.07g, 1.01mmol) and potassium hydroxide (0.113g, 2.02mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.26mL, 4.03mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 480(0.057g, 57%) as a yellow solid.
1H NMR(400MHz，MeOD-d3)δ7.67(d，2H，J＝8.2Hz)，7.39(d，2H，J＝8.2Hz)，7.32-7.15(m，6H)，7.09-7.05(m，1H)，4.88(s，2H)，3.48(s，2H)，3.27(m，4H)，2.30(m，4H)；MS(ESI)m/z 497.2(M++H)。
Example 105: synthesis of Compound 481
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) -4- (2-methoxyphenyl) piperazine-1-formamido) methyl) benzoate)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.20g, 0.436mmol) and 4- (2-methoxyphenyl) piperazine (0.076mL, 0.436mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.157g, 70%) as a yellow oil.
(Compound 481: N- (3-chloro-4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) -4- (2-methoxyphenyl) piperazine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) -4- (2-methoxyphenyl) piperazine-1-carboxamido) methyl) benzoate, 0.10g, 0.195mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.068g, 0.977mmol) and potassium hydroxide (0.109g, 1.95mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.252mL, 3.91mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 481(0.037g, 37%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ7.75(d，2H，J＝8.1Hz)，7.44(d，2H，J＝8.1Hz)，7.41-7.39(m，1H)，7.31-7.19(m，2H)，7.05-6.93(m，4H)，4.96(s，2H)，3.86(s，3H)，3.44(m，4H)，2.92-2.91(m，4H)；MS(ESI)m/z513.1(M++H)。
Example 106: synthesis of Compound 482
(formula 1-4: 4- ((N- (3-chloro-4-fluorophenyl) -4- (4-fluorophenyl) piperazine-1-formamido) methyl) benzoate)
Methyl 4- (((3-chloro-4-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, 0.20g, 0.436mmol) and 1- (4-fluorophenyl) piperazine (0.079mL, 0.436mmol) were dissolved in dimethylformamide (10mL) and the mixture was heated and stirred at 60 ℃ for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-4 (0.184g, 84%) as a yellow oil.
(Compound 482: N- (3-chloro-4-fluorophenyl) -4- (4-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((N- (3-chloro-4-fluorophenyl) -4- (4-fluorophenyl) piperazine-1-carboxamido) methyl) benzoate, 0.10g, 0.20mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine hydrochloride (0.069g, 1.00mmol) and potassium hydroxide (0.112g, 2.00mmol) were added and stirred. Then, hydroxylamine (50 wt% aqueous solution; 0.258mL, 4.00mmol) was added dropwise and stirred at room temperature for 3 hours. After completion of the reaction, methanol was removed under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound 482(0.029g, 29%) as a white solid.
1H NMR(400MHz，MeOD-d3)δ7.75(d，2H，J＝6.8Hz)，7.48-7.43(m，3H)，7.34-7.30(m，1H)，7.22(m，1H)，7.08-6.99(m，4H)，4.98(s，2H)，3.45(m，4H)，3.01(m，4H)；MS(ESI)m/z501.1(M++H)。
Example 107: synthesis of Compound 483
(formula 2-4: 4- ((N- (3- (tetrahydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 2-3 (methyl 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.260g, 0.596mmol) was dissolved in tetrahydrofuran (10mL) at room temperature, Pd/C (10.0%, 0.032g, 0.030mmol) was slowly added, a hydrogen balloon was attached, and the reaction mixture was stirred at the same temperature for 16 hours. The reaction mixture was filtered through a pad of celite, and the filtrate was concentrated under reduced pressure. Then, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 50%) and concentrated to give the desired compound of formula 2-4 (0.217g, 83%) as a white solid.
(Compound 483: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (tetrahydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (tetrahydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 2-4 (0.217g, 0.495mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.51mL, 24.7mmol) was added at room temperature, followed by potassium hydroxide (0.278g, 4.95mmol) and stirred at the same temperature for 30 minutes. Then, the reaction mixture was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous sodium bicarbonate solution (1-2mL) was added and stirred. The precipitated solid was then filtered and dried to give the desired compound 483(0.120g, 55%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，8.97(brs，1H)，7.64(d，2H，J＝8.2Hz)，7.34(d，2H，J＝8.3Hz)，7.23(t，1H，J＝7.7Hz)，6.98-6.94(m，3H)，4.84(s，2H)，3.93-3.90(m，2H)，3.42-3.37(m，6H)，3.12-3.10(m，4H)，2.69(m，1H)，1.62-1.54(m，4H)。MS(ESI)m/z440(M++H)。
Example 108: synthesis of Compound 484
(formula 1-2: 4- (((3-methoxyphenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (m-anisidine; 3.23g, 26.2mmol) and methyl 4- (bromomethyl) benzoate (5.00g, 21.8mmol) were dissolved in acetonitrile (50mL), and N, N-diisopropylethylamine (5.80mL, 32.7mmol) was added and stirred at room temperature for 16 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 5%) and concentrated to give the desired compound of formula 1-2 (5.14g, 87%) as a pale yellow liquid.
(formula 1-3: 4- (((3-methoxyphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- (((3-methoxyphenyl) amino) methyl) benzoate; 5.14g, 18.9mmol) and 4-nitrophenyl chloroformate (4.20g, 20.8mmol) were dissolved in acetonitrile (100mL), followed by addition of potassium carbonate (3.93g, 28.4mmol) and stirring at room temperature for 3 hours. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous sodium bicarbonate, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1 to 3 (5.88g, 71%) as a yellow liquid.
(formula 1-4: 4- ((N- (3-methoxyphenyl) morpholine-4-formamido) methyl) benzoate)
Methyl 4- (((3-methoxyphenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, formula 1-3, 5.88g, 13.5mmol) was dissolved in dimethylformamide (50mL), and morpholine (2.35g, 27.0mmol) and potassium carbonate (5.60g, 40.5mmol) were added and stirred at 60 ℃ for 16 h. After completion of the reaction, the organic layer was extracted with ethyl acetate and saturated aqueous ammonium chloride solution, dehydrated with anhydrous magnesium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and then the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (3.69g, 71%) as a yellow solid.
(Compound 484: N- (4- (hydroxycarbamoyl) benzyl) -N- (3-methoxyphenyl) morpholine-4-carboxamide)
The compound of formula 1-4 (4- ((N- (3-methoxyphenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 0.180g, 0.468mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt.% aqueous solution; 1.43mL, 23.4mmol) was added at room temperature, followed by potassium hydroxide (0.263g, 4.68mmol) and stirred at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous ammonium chloride solution was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was crystallized from dichloromethane (2mL) and hexane (10mL) to give the desired compound 484(0.140g, 78%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.1Hz)，7.32(m，2H)，7.19(t，1H，J＝8.4Hz)，6.69-6.67(m，2H)，6.62(m，1H)，4.84(s，2H)，3.69(s，3H)，3.39-3.36(m，4H)，3.15-3.12(m，4H)。MS(ESI)m/z 386(M++H)。
Example 109: synthesis of Compound 485
(formula 1-4: 4- ((3, 3-difluoro-N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate, 0.22g, 0.46mmol) of the compound of formula 1-3 was dissolved in dimethylformamide (5ml), followed by addition of potassium carbonate (0.19g, 1.37mmol) and 3, 3-difluoroazetidine (0.19g, 0.92 mmol). The mixture was reacted at 60 ℃ for 2 days, diluted with ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compound of formulae 1-4 (0.20g, 101%).
(Compound 485, 3, 3-difluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamide)
Methyl 4- ((3, 3-difluoro-N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamido) methyl) benzoate, 0.20g, 0.47mmol, a compound of formulae 1-4 was dissolved in methanol (10mL), and hydroxylamine (50 wt% aqueous solution; 0.29mL) and potassium hydroxide (0.13g, 2.33mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 485(0.07g, 36%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.84-7.81(m，2H)，7.43-7.34(m，6H)，4.42(s，2H)，4.18-4.14(m，4H)。MS(ESI)m/z 430(M++H)。
Example 110: synthesis of Compound 486
(formula 1-4: 4- ((4-hydroxy-4-phenyl-N- (4- (trifluoromethyl) phenyl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.25g, 0.53mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (5ml), followed by addition of potassium carbonate (0.22g, 1.61mmol) and 4-phenylpiperidin-4-ol (0.19g, 1.07 mmol). The mixture was reacted at 60 ℃ for 2 days, diluted with ethyl acetate and washed with saturated ammonium chloride solution. The organic layer was dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) to give the desired compounds of formulae 1-4 (0.17g, 60%).
(Compound 486, 4-hydroxy-N- (4- (hydroxycarbamoyl) benzyl) -4-phenyl-N- (4- (trifluoromethyl) phenyl) piperidine-1-carboxamide)
Methyl 4- ((4-hydroxy-4-phenyl-N- (4- (trifluoromethyl) phenyl) piperidine-1-carboxamido) methyl) benzoate, 0.17g, 0.32mmol, a compound of formula 1-4 was dissolved in methanol (10mL), and hydroxylamine (50 wt% aqueous solution; 0.20mL) and potassium hydroxide (0.09g, 1.61mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 486(0.09g, 55%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.84-7.82(m，2H)，7.55-7.53(m，2H)，7.43-7.30(m，9H)，4.42(s，2H)，3.39-3.29(m，4H)，2.10-2.05(m，2H)，1.84-1.81(m，2H)。MS(ESI)m/z496(M++H)。
Example 111: synthesis of Compound 487
(formula 6-2: 4- ((3-bromophenylamino) methyl) -3-fluorobenzonitrile)
The compound of formula 6-1 (4- (bromomethyl) -3-fluorobenzonitrile; 1.93g, 9.02mmol) was dissolved in acetonitrile (20mL), and then 3-bromoaniline (1.18mL, 10.8mmol) and N, N-diisopropylethylamine (2.40mL, 13.5mmol) were added at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 6-2 (2.57g, 93%) as a pale yellow liquid.
(formula 6-3: 4- ((3-bromophenylamino) methyl) -3-fluorobenzoic acid)
The compound of formula 6-2 (4- ((3-bromophenylamino) methyl) -3-fluorobenzonitrile; 2.57g, 8.42mmol) and lithium hydroxide monohydrate (3.53g, 84.2mmol) were dissolved in methanol (20 mL)/water (10mL) and stirred at reflux for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. A 1N aqueous hydrochloric acid solution was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain the desired compound of formula 6-3 (2.57g, 94%) as a pale yellow solid.
(formula 6-4; 4- ((3-bromophenylamino) methyl) -3-fluorobenzoic acid methyl ester)
The compound of formula 6-3 (4- ((3-bromophenylamino) methyl) -3-fluorobenzoic acid; 2.57g, 7.93mmol), methanol (6.43mL, 159mmol) and N, N-diisopropylethylamine (4.21mL, 23.8mmol) were dissolved in THF (50mL), then 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC, 3.04g, 15.9mmol) and 1-hydroxybenzotriazole hydrate (HOBt, 2.14g, 15.9mmol) were added at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 6-4 (2.48g, 93%) as a yellow liquid.
(formula 6-5: 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-4 (methyl 4- ((3-bromophenylamino) methyl) -3-fluorobenzoate; 2.48g, 7.33mmol), 4-nitrophenyl chloroformate (1.63g, 8.07mmol) and potassium carbonate (1.52g, 11.0mmol) were dissolved in acetonitrile (30mL) at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain the desired compound of formula 6-5 (3.40g, 92%) as a yellow liquid.
(formula 6-6: 4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-5 (methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) -3-fluorobenzoate; 1.80g, 3.58mmol), morpholine (0.944mL, 10.7mmol) and potassium carbonate (2.47g, 17.9mmol) were dissolved in dimethylformamide (15mL) at 50 ℃ and stirred at the same temperature for 16 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 60%) and concentrated to give the desired compound of formula 6-6 (1.58g, 98%) as a yellow liquid.
(Compound 487: N- (3-bromophenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
The compound of formula 6-6 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester; 0.200g, 0.443mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.90mL, 31.0mmol) was added at room temperature, followed by potassium hydroxide (0.249g, 4.43mmol) and stirred at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (2mL) and diethyl ether (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 487(0.162g, 81%) as a pale brown solid.
1H NMR(400MHz，DMSO-d6)δ11.23(brs，1H)，9.14(brs，1H)，7.54-7.45(m，3H)，7.40(s，1H)，7.31-7.27(m，2H)，7.15(m，1H)，4.88(s，2H)，3.41-3.38(m，4H)，3.14-3.12(m，4H)。MS(ESI)m/z 452，454(M++H)。
Example 112: synthesis of Compound 488
(formula 6-6: 4- ((N- (3-bromophenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-5 (methyl 4- (((3-bromophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) -3-fluorobenzoate; 1.60g, 3.18mmol), 4-hydroxypiperidine (0.965g, 9.54mmol) and potassium carbonate (2.20g, 15.9mmol) were dissolved in dimethylformamide (15mL) at 50 ℃ and stirred at the same temperature for 16 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 80%) and concentrated to give the desired compound of formula 6-6 (1.16g, 78%) as a yellow liquid.
(Compound 488: N- (3-bromophenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -4-hydroxypiperidine-1-carboxamide)
Methyl 4- ((N- (3-bromophenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoate of the formula 6-6 (0.200g, 0.430mmol) was dissolved in methanol (10mL), and hydroxylamine (50.0 wt% aqueous solution; 1.84mL, 30.1mmol) was added at room temperature, followed by potassium hydroxide (0.241g, 4.30mmol) and stirring at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (2mL) and diethyl ether (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 488(0.101g, 50%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.23(brs，1H)，9.14(brs，1H)，7.63-7.46(m，3H)，7.32(s，1H)，7.27-7.24(m，2H)，6.90(m，1H)，4.85(s，2H)，4.68(s，1H)，3.55-3.45(m，3H)，2.87-2.82(m，2H)，1.56-1.53(m，2H)，1.15-1.12(m，2H)。MS(ESI)m/z 466，468(M++H)。
Example 113: synthesis of Compound 489
(Compound 489: 4- (3- (N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamido) phenyl) piperidine-1-carboxylic acid tert-butyl ester)
The compound of formula 4-4 (tert-butyl 4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) piperidine-1-carboxylate; 0.100g, 0.186mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.14mL, 18.6mmol) was added at room temperature, followed by potassium hydroxide (0.104g, 1.86mmol) and stirring at the same temperature for 30 minutes. Then, the reaction mixture was concentrated under reduced pressure until its volume became about 1mL, and a saturated aqueous sodium bicarbonate solution (1mL) was added and stirred. The precipitated solid was then filtered and dried to give the desired compound 489(0.086g, 86%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.4Hz)，7.34(d，2H，J＝8.3Hz)，7.22(t，1H，J＝7.6Hz)，6.99-6.93(m，3H)，4.84(s，2H)，4.06-4.02(m，2H)，3.40-3.37(m，4H)，3.12-3.10(m，4H)，2.66-2.63(m，2H)，1.71-1.68(m，2H)，1.44(m，3H)，1.40(s，9H)。MS(ESI)m/z539(M++H)。
Example 114: synthesis of Compound 490
(formula 4-4: 4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) piperidine-1-carboxylic acid tert-butyl ester)
The compound of formula 4-1 (4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester; 3.28g, 6.12mmol) was dissolved in THF (30mL) at room temperature, Pd/C (130mg) was slowly added, a hydrogen balloon was attached, and the reaction mixture was stirred at the same temperature for 16 hours. The reaction mixture was filtered through a celite pad, and the filtrate was concentrated under reduced pressure to obtain the desired compound of formula 4-4 (3.28g, 99%) as a white solid.
(formula 4-5: 4- ((N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride)
The compound of formula 4-4 (4- (3- (N- (4- (methoxycarbonyl) benzyl) morpholine-4-carboxamido) phenyl) piperidine-1-carboxylic acid tert-butyl ester; 3.10g, 5.77mmol) was dissolved in 1, 4-bisAlkane (10mL) was then added hydrogen chloride (4.0M 1, 4-bis) at room temperatureAn alkane solution; 14.4mL, 57.7mmol) and stirred at the same temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to give the desired compound of formula 4-5 (2.65g, 99%) as a white solid.
(formula 4-6: 4- ((N- (3- (1-acetylpiperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride of the compound of the formula 4-5 (0.150g, 0.316mmol) and N, N-diisopropylethylamine (0.168mL, 0.949mmol) were dissolved in dichloromethane (3mL), followed by addition of acetic anhydride (0.065g, 0.633mmol) at room temperature and stirring at the same temperature for 1 hour. Saturated aqueous sodium bicarbonate was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 80 to 100%) and concentrated to give the desired compound of formulae 4 to 6 (0.148g, 98%) as a colorless liquid.
(Compound 490: N- (3- (1-acetylpiperidin-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1-acetylpiperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-6 (0.148g, 0.309mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.32mL, 21.6mmol) was added at room temperature, followed by potassium hydroxide (0.173g, 3.09mmol) and stirring for 30 minutes at the same time. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (5mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 490(0.036g, 24%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，9.03(brs，1H)，7.63(d，2H，J＝8.4Hz)，7.33(d，2H，J＝8.4Hz)，7.22(t，1H，J＝7.9Hz)，6.99-6.93(m，3H)，4.84(s，2H)，4.50(m，1H)，3.89(m，1H)，3.39-3.36(m，4H)，3.12-3.10(m，4H)，2.68(m，1H)，2.57-2.53(m，2H)，2.01(s，3H)，1.74-1.70(m，2H)，1.54(m，1H)，1.38(m，1H)。MS(ESI)m/z 481(M++H)。
Example 115: synthesis of Compound 491
(formula 4-6: 4- ((N- (3- (1- (methylsulfonyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride of formula 4-5 (0.150g, 0.316mmol) and N, N-diisopropylethylamine (0.168mL, 0.949mmol) were dissolved in dichloromethane (3mL), followed by addition of methanesulfonyl chloride (0.049mL, 0.633mmol) at room temperature and stirring at the same temperature for 1 hour. Saturated aqueous sodium bicarbonate was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 100%) and concentrated to give the desired compound of formulae 4 to 6 (0.119g, 73%) as a colorless liquid.
(Compound 491: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1- (methylsulfonyl) piperidin-4-yl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (1- (methylsulfonyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-6 (0.119g, 0.231mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 0.988mL, 16.2mmol) was added at room temperature, followed by potassium hydroxide (0.129g, 2.31mmol) and stirring for 30 minutes at the same time. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (5mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 491(0.043g, 36%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.64(d，2H，J＝8.2Hz)，7.34(d，2H，J＝8.3Hz)，7.23(t，1H，J＝7.8Hz)，7.02(s，1H)，6.97(d，2H，J＝7.9Hz)，4.85(s，2H)，3.66-3.63(m，2H)，3.38-3.36(m，4H)，3.28-3.26(m，2H)，3.12-3.10(m，4H)，2.79-2.76(m，2H)，2.66-2.61(m，2H)，1.82-1.79(m，2H)，1.64-1.60(m，2H)。MS(ESI)m/z 517(M++H)。
Example 116: synthesis of Compound 492
(formula 4-6: 4- ((N- (3- (1- (isopropylcarbamoyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride of formula 4-5 (0.150g, 0.316mmol) and N, N-diisopropylethylamine (0.168mL, 0.949mmol) were dissolved in dichloromethane (3mL), followed by addition of isopropyl isocyanate (0.054mL, 0.633mmol) at room temperature and stirring at the same temperature for 1 hour. Saturated aqueous sodium bicarbonate was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (, silica; ethyl acetate/hexane ═ 50 to 100%) and concentrated to give the desired compound of formulae 4 to 6 (0.101g, 61%) as a white solid.
(Compound 492: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (1- (isopropylcarbamoyl) piperidin-4-yl) phenyl) morpholine-4-carboxamide)
A compound of formula 4-6 (methyl 4- ((N- (3- (1- (isopropylcarbamoyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.101g, 0.193mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 0.827mL, 13.5mmol) was added at room temperature, followed by potassium hydroxide (0.108g, 1.93mmol) and stirring at the same time for 30 minutes. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (5mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 492(0.047g, 46%) as a light brown solid.
1H NMR(400MHz，DMSO-d6)δ7.63(d，2H，J＝8.2Hz)，7.34(d，2H，J＝8.2Hz)，7.22(t，1H，J＝8.2Hz)，6.97-6.92(m，3H)，6.13(d，1H，J＝7.5Hz)，4.84(s，2H)，4.08-4.05(m，2H)，3.76(m，1H)，3.34(m，4H)，3.11(m，4H)，2.69-2.63(m，3H)，1.67-1.64(m，2H)，1.47-1.42(m，2H)，1.04(d，6H，J＝6.6Hz).MS(ESI)m/z 524(M++H)。
Example 117: synthesis of Compound 493
(Compound 493: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamide hydrochloride)
Compound 489 (tert-butyl 4- (3- (N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamido) phenyl) piperidine-1-carboxylate; 0.050g, 0.093mmol) was dissolved in 1, 4-bisAlkane (3mL) was then added hydrogen chloride (4.0M 1, 4-bis) at room temperatureAn alkane solution; 0.696mL, 2.79mmol) and stirred at the same temperature, and the reaction mixture was concentrated under reduced pressure. Diethyl ether (10mL) was added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 493(0.027g, 61%) as a brown solid.
1H NMR(400MHz，DMSO-d6)δ11.16(brs，1H)，9.04(brs，1H)，8.88(brs，1H)，7.64(d，2H，J＝7.8Hz)，7.37-7.24(m，3H)，7.12-6.98(m，2H)，4.87(s，2H)，3.26(m，3H)，3.11(m，4H)，2.94(m，2H)，2.82(m，2H)，1.83-1.78(m，3H)，1.39(m，4H)。MS(ESI)m/z 439(M++H)。
Example 118: synthesis of Compound 494
(formula 1-4: 4- ((4-acetyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
The compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.21g, 0.45mmol) was dissolved in dimethylformamide (3mL), followed by the addition of potassium carbonate (0.19g, 1.34mmol) and 1- (piperazin-1-yl) ethanone (0.06g, 0.45 mmol). The mixture was reacted at 60 ℃ for 1 day and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 60%) to give the desired compounds of formulae 1-4 (0.16g, 75%).
(Compound 494: 4-acetyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4-acetyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.16g, 0.33mmol, of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.21mL) and potassium hydroxide (0.09g, 1.67mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and water, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 494(0.02g, 10%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.16(brs，1H)，9.17(brs，1H)，7.65(d，2H，J＝8.2Hz)，7.53-7.36(m，6H)，4.95(s，2H)，3.29(m，4H)，3.15(m，4H)，1.94(s，3H)。MS(ESI)m/z 465(M++H)。
Example 119: synthesis of Compound 495
(formula 1-4 (R) -4- ((3-fluoro-N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate, 0.23g, 0.49mmol) of the compound of formula 1-3 was dissolved in dimethylformamide (5mL), followed by addition of potassium carbonate (0.20g, 1.48mmol) and (R) -3-fluoropyrrolidine hydrochloride (0.12g, 0.99 mmol). The mixture was reacted at 60 ℃ for 1 day and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 40%) to give the desired compounds of formulae 1-4 (0.18g, 87%).
(Compound 495 (R) -3-fluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((R) -4- ((3-fluoro-N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.18g, 0.43mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.26mL) and potassium hydroxide (0.12g, 2.16mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and saturated aqueous potassium carbonate solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 495(0.10g, 54%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(brs，1H)，9.00(brs，1H)，7.64(d，2H，J＝7.7Hz)，7.52(t，1H，J＝7.9Hz)，7.45-7.35(m，5H)，5.19(d，1H，J＝53.1Hz)，4.99-4.89(m，2H)，3.32-3.18(m，3H)，3.09-3.02(m，1H)，1.99-1.83(m，2H)。MS(ESI)m/z 426(M++H)。
Example 120: synthesis of Compound 496
(formula 1-4 (S) -4- ((3-fluoro-N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate, 0.29g, 0.62mmol) of the compound of formula 1-3 was dissolved in dimethylformamide (5mL), followed by addition of potassium carbonate (0.26g, 1.86mmol) and (S) -3-fluoropyrrolidine hydrochloride (0.16g, 1.24 mmol). The mixture was reacted at 60 ℃ for 1 day and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 40%) to give the desired compounds of formulae 1-4 (0.20g, 75%).
(Compound 496 (S) -3-fluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((S) -4- ((3-fluoro-N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.20g, 0.47mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.29mL) and potassium hydroxide (0.13g, 2.33mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and saturated aqueous potassium carbonate solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 496(0.14g, 72%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.15(brs，1H)，10.18(brs，1H)，9.00(brs，1H)，7.65(d，2H，J＝8.2Hz)，7.53-7.49(m，1H)，7.45-7.33(m，5H)，5.19(d，1H，J＝53.2Hz)，4.99-4.89(m，2H)，3.36-3.19(m，3H)，3.09-3.02(m，1H)，1.99-1.83(m，2H)。MS(ESI)m/z 426(M++H)。
Example 121: synthesis of Compound 497
(formula 1-4 (R) -4- ((2- (hydroxymethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.23g, 0.48mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (5mL), followed by addition of potassium carbonate (0.20g, 1.44mmol) and (R) -pyrrolidin-2-ylmethanol (0.10g, 0.96 mmol). The mixture was reacted at 60 ℃ for 1 day and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 40%) to give the desired compounds of formulae 1-4 (0.15g, 73%).
(Compound 497 (R) -N- (4- (hydroxycarbamoyl) benzyl) -2- (hydroxymethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((R) -4- ((2- (hydroxymethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.15g, 0.39mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.21mL) and potassium hydroxide (0.10g, 1.74mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and saturated aqueous potassium carbonate solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 497(0.04g, 28%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.84-7.82(m，3H)，7.65-7.63(m，1H)，7.41-7.17(m，4H)，4.42(s，2H)，3.88-3.63(m，2H)，3.43-3.30(m，3H)，1.67-1.42(m，4H)。MS(ESI)m/z 438(M++H)。
Example 122: synthesis of Compound 498
(formula 1-4 (S) -4- ((2- (trifluoromethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.22g, 0.47mmol) of the compound of the formula 1-3 was dissolved in dimethylformamide (5mL), followed by addition of potassium carbonate (0.20g, 1.42mmol) and (S) -2- (trifluoromethyl) pyrrolidine (0.13g, 0.95 mmol). The mixture was reacted at 60 ℃ for 1 day and then diluted with a saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) to give the desired compounds of formulae 1-4 (0.17g, 74%).
(Compound 498 (S) -N- (4- (hydroxycarbamoyl) benzyl) -2- (trifluoromethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((S) -4- ((2- (trifluoromethyl) -N- (3- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.17g, 0.35mmol) of the compound of formula 1-4 was dissolved in methanol (5mL), and hydroxylamine (50 wt% aqueous solution; 0.21mL) and potassium hydroxide (0.10g, 1.74mmol) were added and stirred overnight. After completion of the reaction, methanol was removed by distillation under reduced pressure, and the organic layer was extracted with ethyl acetate and saturated aqueous potassium carbonate solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) to give the desired compound 498(0.07g, 44%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.13(brs，1H)，8.98(brs，1H)，7.84-7.82(m，3H)，7.65-7.63(m，1H)，7.41-7.17(m，4H)，4.42(s，2H)，4.17-4.15(m，1H)，3.40-3.30(m，2H)，1.67-1.42(m，4H)。MS(ESI)m/z438(M++H)。
Example 123: synthesis of Compound 499
(formula 10-2: 4- ((3- (hydroxymethyl) phenylamino) methyl) benzoic acid methyl ester)
The compound of formula 10-1 (3-hydroxymethylaniline; 5.00g, 40.6mmol), methyl 4-formylbenzoate (8.00g, 48.7mmol) and acetic acid (4.64mL, 81.2mmol) were dissolved in methanol (100mL) and stirred at room temperature for 30 minutes, followed by addition of sodium cyanoborohydride (3.83g, 60.9mmol) and stirring at the same temperature for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 50%) and concentrated to give the desired compound of formula 10-2 (10.9g, 99%) as a pale yellow solid.
(formula 10-3: 4- (((3- (hydroxymethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 10-2 (methyl 4- ((3- (hydroxymethyl) phenylamino) methyl) benzoate; 1.60g, 5.90mmol) and potassium carbonate (1.63g, 11.8mmol) were dissolved in acetonitrile (100mL), followed by addition of 4-nitrophenyl chloroformate (1.31g, 6.49mmol) at room temperature and stirring at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain the desired compound of formula 10-3 (2.43g, 94%) as a yellow liquid.
(formula 10-4: 4- ((N- (3- (hydroxymethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 10-3 (methyl 4- (((3- (hydroxymethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 2.43g, 5.57mmol), morpholine (2.45mL, 27.8mmol) and potassium carbonate (3.85g, 27.8mmol) were dissolved in dimethylformamide (20mL) at room temperature and stirred at the same temperature for 60 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 100%) and concentrated to give the desired compound of formula 10-4 (1.96g, 92%) as a yellow liquid.
(formula 10-5: 4- ((N- (3- (hydroxymethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid)
A compound of formula 10-4 (4- ((N- (3- (hydroxymethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester; 1.96g, 5.10mmol) and lithium hydroxide monohydrate (2.14g, 51.0mmol) were dissolved in methanol (10 mL)/water (5mL) and stirred at 50 ℃ for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. 0.5N aqueous hydrochloric acid solution was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain the desired compound of formula 10-5 (1.82g, 96%) as a pale yellow solid.
(formula 10-6: N- (3- (hydroxymethyl) phenyl) -N- (4- (tetrahydro-2H-pyran-2-yloxycarbamoyl) benzyl) morpholine-4-carboxamide)
A compound of formula 10-5 (4- ((N- (3- (hydroxymethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid; 1.82g, 4.91mmol) and N, N-diisopropylethylamine (2.61mL, 14.7mmol) were dissolved in dichloromethane (30mL) and O- (tetrahydropyran-2-yl) hydroxylamine (0.748g, 6.39mmol) was added at room temperature. Next, 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC, 1.88g, 9.83mmol) and 1-hydroxybenzotriazole hydrate (HOBt, 1.34g, 9.83mmol) were added and then stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 70-100%) and concentrated to give the desired compound of formula 10-6 (1.35g, 58%) as a white solid.
(Compound 499: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (hydroxymethyl) phenyl) morpholine-4-carboxamide)
The compound of formula 10-6 (N- (3- (hydroxymethyl) phenyl) -N- (4- (tetrahydro-2H-pyran-2-yloxycarbamoyl) benzyl) morpholine-4-carboxamide; 0.050g, 0.106mmol) was dissolved in methanol (3mL) and then hydrogen chloride (4.0M 1, 4-bis-formamide) was added at room temperatureAn alkane solution; 0.799mL, 3.20mmol) and stirred at the same temperature for 1 hour. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain the desired compound 499(0.016g, 39%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，8.97(brs，1H)，7.64(d，2H，J＝8.2Hz)，7.35(d，2H，J＝8.2Hz)，7.27(m，1H)，7.10(s，1H)，6.99-6.97(m，2H)，4.88(s，2H)，4.43(s，2H)，3.38(m，4H)，3.16(m，4H)。MS(ESI)m/z 386(M++H)。
Example 124: synthesis of Compound 500
(formula 10-7: 4- ((N- (3- (fluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 10-5 (4- ((N- (3- (hydroxymethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid; 1.25g, 3.25mmol) was dissolved in dichloromethane (20mL) and diethylaminosulfur trifluoride (DAST, 0.424mL, 3.58mmol) was added at 0 ℃ and stirred at the same temperature for 1 hour. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 50%) and concentrated to give the desired compound of formula 10-7 (0.617g, 49%) as a colorless liquid.
(Compound 500: N- (3- (fluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
A compound of formula 10-7 (methyl 4- ((N- (3- (fluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.100g, 0.259mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.11mL, 18.1mmol) was added at room temperature, followed by potassium hydroxide (0.145g, 2.59mmol) and stirred at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (5mL) and hexane (30mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 500(0.089g, 89%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，8.98(brs，1H)，7.64(d，2H，J＝8.3Hz)，7.36-7.32(m，3H)，7.20(s，1H)，7.15(d，1H，J＝7.5Hz)，7.09(d，1H，J＝7.4Hz)，5.36(d，2H，J＝47.5Hz)，4.87(s，2H)，3.39(t，4H，J＝4.6Hz)，3.13(t，4H，J＝4.6Hz).MS(ESI)m/z 388(M++H)。
Example 125: synthesis of Compound 511
(formula 6-7: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-6 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester; 0.200g, 0.443mmol), benzo [ d ] b][1，3]Dioxol-5-ylboronic acid (0.096g, 0.576mmol), Pd (dppf) Cl2(0.018g, 0.022mmol) and sodium carbonate (2.0M aqueous solution; 0.665mL, 1.33mmol) were dissolved in 1, 4-bisAlkane (2mL) and stirred at 110 ℃ for 16 h. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30-60%) and concentrated to give the desired compound of formulae 6-7 (0.077g, 35%) as a white solid.
(Compound 511: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoate of the compound of formula 6-7 (0.077g, 0.156mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt.% in water; 0.956mL, 15.6mmol) was added at room temperature, followed by potassium hydroxide (0.088g, 1.56mmol) and stirring at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate solution was poured into the obtained concentrate and stirred. Then, the precipitated solid was filtered and dried to obtain the desired compound 511(0.056g, 73%) as a white solid.
MS(ESI)m/z 494(M++H)。
Example 126: synthesis of Compound 512
(formula 6-7: 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-6 (4- ((N- (3-bromophenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoic acid methyl ester; 0.300g, 0.665mmol), 3, 6-dihydro-2H-pyran-4-boronic acid pinacol ester (0.182g, 0.864mmol), Pd (dppf) Cl2(0.027g, 0.033mmol) and sodium carbonate (2.0M aqueous solution; 0.997mL, 1.99mmol) were dissolved in 1, 4-bisAlkane (3mL) and stirred at 110 ℃ for 16 h. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20-50%) and concentrated to give the desired compound of formulae 6-7 (0.068g, 23%) as a pale brown solid.
(Compound 512: N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) morpholine-4-carboxamido) methyl) -3-fluorobenzoate of formula 6-7 (0.068g, 0.150mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 0.915mL, 15.0mmol) was added at room temperature, followed by potassium hydroxide (0.084g, 1.50mmol) and stirring at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate solution was poured into the obtained concentrate and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (3mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 512(0.028g, 41%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.25(brs，1H)，9.11(brs，1H)，7.53-7.43(m，3H)，7.29(m，1H)，7.21-7.17(m，2H)，7.04(m，1H)，6.25(s，1H)，4.88(s，2H)，4.20(m，2H)，3.79(m，2H)，3.36(m，4H)，3.12(m，4H)，2.38(m，2H)。MS(ESI)m/z 456(M++H)。
Example 127: synthesis of Compound 513
(formula 6-7: 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-6 (methyl 4- ((N- (3-bromophenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoate; 0.200g, 0.430mmol), benzo [ d][1，3]Dioxol-5-ylboronic acid (0.093g, 0.559mmol), Pd (dppf) Cl2(0.018g, 0.021mmol) and sodium carbonate (2.0M aqueous solution; 0.645mL, 1.29mmol) were dissolved in 1, 4-bisAlkane (2mL) and stirred at 110 ℃ for 16 h. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 40 to 70%) and concentrated to give the desired compound of formulae 6 to 7 (0.037g, 17%) as a pale yellow solid.
(Compound 513: N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -4-hydroxypiperidine-1-carboxamide)
A compound of formula 6-7 (methyl 4- ((N- (3- (benzo [ d ] [1, 3] dioxol-5-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoate; 0.037g, 0.073mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 0.447mL, 7.31mmol) was added at room temperature, followed by potassium hydroxide (0.041g, 0.730mmol) and stirring at the same temperature for 30 minutes. Saturated aqueous sodium bicarbonate solution was poured into the obtained concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 513(0.032g, 86%) as a beige solid.
1H NMR(400MHz，DMSO-d6)δ7.63-7.59(m，2H)，7.54(m，1H)，7.36-7.32(m，3H)，7.21(m，1H)，7.09(m，1H)，7.02(m，1H)，6.96(m，1H)，6.65(s，2H)，4.89(s，2H)，4.63(d，1H，J＝4.0Hz)，3.52-3.49(m，3H)，2.85-2.82(m，2H)，1.53-1.50(m，2H)，1.13-1.09(m，2H)。MS(ESI)m/z 508(M++H)。
Example 128: synthesis of Compound 514
(formula 6-7: 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoic acid methyl ester)
A compound of formula 6-6 (methyl 4- ((N- (3-bromophenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoate, 0.300g, 0.645mmol), 3, 6-dihydro-2H-pyran-4-boronic acid pinacol ester (0.176g, 0.838mmol), Pd (dppf) Cl2(0.026g, 0.032mmol) and sodium carbonate (2.0M aqueous solution; 0.967mL, 1.93mmol) were dissolved in 1, 4-bisAlkane (3mL) and stirred at 110 ℃ for 16 h. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue is purified by column chromatography (silica; ethyl acetate/hexane)Alkane-50-80%) and concentrated to give the desired compound of formula 6-7 (0.078g, 26%) as a light yellow solid.
(Compound 514: N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) -N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -4-hydroxypiperidine-1-carboxamide)
Methyl 4- ((N- (3- (3, 6-dihydro-2H-pyran-4-yl) phenyl) -4-hydroxypiperidine-1-carboxamido) methyl) -3-fluorobenzoate of the formula 6-7 (0.078g, 0.166mmol) was dissolved in methanol (10mL), hydroxylamine (50.0% by weight aqueous solution; 1.02mL, 16.6mmol) was added at room temperature, followed by potassium hydroxide (0.093g, 1.67mmol) and stirring at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate solution was poured into the obtained concentrate and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (3mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 514(0.036g, 46%) as a pale green solid.
1H NMR(400MHz，DMSO-d6)δ11.25(brs，1H)，9.11(brs，1H)，7.54-7.44(m，3H)，7.28(t，1H，J＝7.8Hz)，7.18-7.15(m，2H)，7.00(d，1H，J＝8.6Hz)，6.23(s，1H)，4.84(s，2H)，4.63(d，1H，J＝4.0Hz)，4.19(m，2H)，3.79(t，2H，J＝5.4Hz)，3.52-3.46(m，3H)，2.83-2.76(m，2H)，2.37(m，2H)，1.52-1.49(m，2H)，1.16-1.13(m，2H)。MS(ESI)m/z 470(M++H)。
Example 129: synthesis of Compound 517
(formula 4-7: 4- ((N- (3- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid ethyl ester)
A compound of formula 4-5 (methyl 4- ((N- (3- (piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate hydrochloride, 0.540g, 1.139mmol), 2, 2-dimethyloxirane (0.411g, 5.696mmol) and potassium carbonate (0.472g, 3.42mmol) were combined with water (1 mL)/ethanol (3 mL). The mixture was then heated at 120 ℃ for 20 minutes under microwave irradiation, and then the temperature was lowered to room temperature. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Water was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 2 to 10%) and concentrated to give the desired compound of formulae 4 to 7 (0.325g, 55%) as a colorless liquid.
(Compound 517: N- (3- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Ethyl 4- ((N- (3- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-7 (0.050 g, 0.095mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 0.584mL, 9.55mmol) was added at room temperature, followed by potassium hydroxide (0.054g, 0.955mmol) and stirring at the same temperature for 30 minutes. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (2mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 517(0.032g, 66%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，8.98(brs，1H)，7.63(d，2H，J＝8.2Hz)，7.34(d，2H，J＝8.1Hz)，7.21(m，1H)，6.97-6.93(m，3H)，4.83(s，2H)，3.37-3.34(m，6H)，3.12-3.10(m，4H)，3.00(m，1H)，2.32(m，1H)，2.19-2.14(m，3H)，1.68-1.62(m，4H)，1.22(s，3H)，1.10(m，4H)。MS(ESI)m/z 511(M++H)。
Example 130: synthesis of Compound 518
(formula 4-8: 4- ((N- (3- (1- (2-fluoro-2-methylpropyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid ethyl ester)
Ethyl 4- ((N- (3- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-7 (0.275 g, 0.525mmol) was dissolved in dichloromethane (20mL), and diethylaminosulfur trifluoride (DAST, 0.068mL, 0.578mmol) was added at 0 ℃ and stirred at the same temperature for 1 hour. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; methanol/dichloromethane ═ 2 to 10%) and concentrated to give the desired compounds of formulae 4 to 8 (0.146g, 53%) as colorless liquids.
(Compound 518: N- (3- (1- (2-fluoro-2-methylpropyl) piperidin-4-yl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Ethyl 4- ((N- (3- (1- (2-fluoro-2-methylpropyl) piperidin-4-yl) phenyl) morpholine-4-carboxamido) methyl) benzoate, a compound of formula 4-8 (0.146g, 0.278mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt% aqueous solution; 1.19mL, 19.4mmol) was added at room temperature, followed by potassium hydroxide (0.156g, 2.78mmol) and stirring at the same temperature. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane (2mL) and hexane (20mL) were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 518(0.092g, 65%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.12(brs，1H)，8.99(brs，1H)，7.64(d，2H，J＝8.3Hz)，7.34(d，2H，J＝8.2Hz)，7.21(t，1H，J＝7.7Hz)，6.98-6.94(m，3H)，4.84(s，2H)，3.37-3.35(m，4H)，3.14-3.10(m，4H)，2.97-2.95(m，2H)，2.46-2.38(m，3H)，2.17-2.14(m，2H)，1.67-1.60(m，4H)，1.33(s，3H)，1.28(s，3H)。MS(ESI)m/z 513(M++H)。
Example 131: synthesis of Compound 520
(formula 1-4 (R) -4- ((3-fluoro-N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.239g, 0.504mmol), (R) -3-fluoropyrrolidine hydrochloride (0.127g, 1.008mmol) and potassium carbonate (0.209g, 1.513mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 24 hours, after which the reaction was completed while the temperature was lowered to room temperature. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 40%) and concentrated to give the desired compound of formulae 1-4 (0.081g, 37.8%) as a white solid.
(Compound 520, (R) -3-fluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((R) -4- ((3-fluoro-N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.081g, 0.191mmol) and hydroxylamine (50% by weight in water; 0.063g, 1.909mmol) of the compound of formula 1-4 were dissolved in methanol (5ml) and stirred at room temperature for 24 hours. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 520(0.067g, 83.0%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(s，1H)，8.99(s，1H)，7.66-7.63(m，4H)，7.37(d，2H，J＝8.1Hz)，7.30(d，2H，J＝8.4Hz)，5.27(s，0.5H)，5.14(s，0.5H)，4.98-4.88(m，2H)，3.34-3.30(m，2H)，3.18-3.11(m，2H)，2.02-1.92(m，2H)；MS(ESI)m/z 426.1(M++H)。
Example 132: synthesis of Compound 521
(formula 1-4 (S) -4- ((3-fluoro-N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.312g, 0.659mmol), (S) -3-fluoropyrrolidine hydrochloride (0.165g, 1.317mmol) and potassium carbonate (0.273g, 1.976mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 24 hours, after which the reaction was completed while the temperature was lowered to room temperature. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 40%) and concentrated to give the desired compound of formulae 1 to 4 (0.115g, 41.0%) as a white solid.
(Compound 521, (S) -3-fluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl ((S) -4- ((3-fluoro-N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.115g, 0.271mmol) and hydroxylamine (50% by weight in water; 0.090g, 2.710mmol) of the compound of formula 1-4 were dissolved in methanol (5ml) and stirred at room temperature for 24 hours. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 521(0.075g, 65.2%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(s，1H)，9.00(s，1H)，7.66-7.63(m，4H)，7.37(d，2H，J＝8.1Hz)，7.30(d，2H，J＝8.4Hz)，5.27(s，0.5H)，5.14(s，0.5H)，5.02-4.88(m，2H)，3.34-3.30(m，2H)，3.24-3.11(m，2H)，2.07-1.86(m，2H)；MS(ESI)m/z 426.1(M++H)。
Example 133: synthesis of Compound 522
(formula 1-4 (R) -4- ((2- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.312g, 0.659mmol), (R) -pyrrolidin-2-ylmethanol (0.133g, 1.317mmol) and potassium carbonate (0.273g, 1.976mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 24 hours, after which the reaction was completed while the temperature was lowered to room temperature. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 1 to 4 (0.116g, 40.4%) as a yellow liquid.
(Compound 522, (R) -N- (4- (hydroxycarbamoyl) benzyl) -2- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamide)
Methyl (((R) -4- ((2- (hydroxymethyl) -N- (4- (trifluoromethyl) phenyl) pyrrolidine-1-carboxamido) methyl) benzoate; 0.116g, 0.273mmol) and hydroxylamine (50% by weight in water; 0.088g, 2.658mmol) were dissolved in methanol (5ml) and stirred at room temperature for 24 hours the concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure, saturated aqueous sodium bicarbonate solution was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate, the organic layer was washed with saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure, the concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to obtain the desired compound 522(0.025g, 20.7%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(s，1H)，8.99(s，1H)，7.65(d，2H，J＝8.3Hz)，7.59(d，2H，J＝8.6Hz)，7.38(d，2H，J＝8.2Hz)，7.32(d，2H，J＝8.5Hz)，5.07(s，0.5H)，5.03(s，0.5H)，4.85-4.79(m，2H)，3.99-3.94(m，1H)，3.60-3.50(m，2H)，3.12-3.07(m，1H)，2.67-2.59(m，1H)，1.89-1.83(m，1H)，1.75-1.66(m，2H)，1.62-1.60(m，1H)；MS(ESI)m/z 438.2(M++H)。
Example 134: synthesis of Compound 529
(formula 1-2: 4- (((3-fluorophenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (3-fluoroaniline; 0.865mL, 8.999mmol) and methyl 4-formylbenzoate (1.477g, 8.999mmol) were dissolved in methanol (50mL) and stirred at room temperature for 3 hours. Then, acetic acid (1.029mL, 17.999mmol) and sodium cyanoborohydride (95.0%, 0.595g, 8.999mmol) were added and stirred at the same temperature for a further 24 hours. Saturated aqueous sodium bicarbonate was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 20%) and concentrated to give the desired compound of formula 1-2 (1.840g, 78.9%) as a colorless liquid.
(formula 1-3: 4- (((3-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((3-fluorophenyl) amino) methyl) benzoate; 2.700g, 10.413mmol), 4-nitrophenyl chloroformate (4.198g, 20.827mmol) and potassium carbonate (4.318g, 31.240mmol) were dissolved in acetonitrile (100mL) at room temperature and stirred at the same temperature for 24 hours. Saturated aqueous sodium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 0 to 20%) and concentrated to give the desired compound of formulae 1 to 3 (2.650g, 60.0%) as a colorless liquid.
(formula 1-4: 4- ((4-acetyl-N- (3-fluorophenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
A compound of formula 1-3 (methyl 4- (((3-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.247g, 0.582mmol), 1- (piperazin-1-yl) ethanone (0.145mL, 1.164mmol) and potassium carbonate (0.241g, 1.746mmol) were dissolved in N, N-dimethylformamide (5mL) at 60 ℃ and stirred at the same temperature for 16 hours. Saturated aqueous sodium bicarbonate was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 40 to 100%) and concentrated to give the desired compound of formulae 1 to 4 (0.112g, 46.7%) as a light yellow liquid.
(Compound 529: 4-acetyl-N- (3-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((4-acetyl-N- (3-fluorophenyl) piperazine-1-carboxamido) methyl) benzoate, 0.112g, 0.271mmol and hydroxylamine (50.0 wt.% in water, 0.166mL, 2.709mmol) were dissolved in methanol (10mL), followed by addition of potassium hydroxide (0.076g, 1.354mmol) at room temperature and stirring at the same temperature for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was dehydrated with anhydrous magnesium sulfate and then concentrated under reduced pressure. The precipitated solid was filtered and dried to give the desired compound 529(0.048g, 42.8%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.1(s，1H)，8.99(s，1H)，7.64(d，2H，J＝8.2Hz)，7.37(d，2H，J＝8.2Hz)，7.33-7.28(m，1H)，7.07-7.02(m，1H)，6.99-6.94(m，1H)，6.89-6.84(m，1H)，4.89(s，2H)，3.30-3.29(m，2H)，3.20-3.15(m，4H)，2.55-2.54(m，1H)，2.45-2.44(m，1H)，1.94(s，3H)；MS(ESI)m/z 415.1(M++H)。
Example 135: synthesis of Compound 530
(formula 1-2: 4- ((3-fluorophenylamino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (methyl 4-formylbenzoate; 1.47g, 8.99mmol) was dissolved in methanol (50mL), followed by addition of 3-fluoroaniline (1.0g, 8.99 mmol). The mixture was allowed to react at room temperature for 3 hours, then sodium cyanoborohydride (NaCNBH) was added3(ii) a 0.56g, 8.99mmol) and acetic acid (1.03mL, 17.99 mmol). After the mixture was reacted at room temperature for 1 day, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was dehydrated with anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formula 1-2 (1.84g, 79%).
(formula 1-3: 4- (((3-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-2 (methyl 4- ((3-fluorophenylamino) methyl) benzoate; 2.7g, 10.4mmol) and 4-nitrophenylchloroformate (4.20g, 20.8mmol) were dissolved in acetonitrile (100mL), followed by addition of potassium carbonate (4.32g, 31.2 mmol). The mixture was reacted at room temperature for 1 day, and then diluted with ethyl acetate. The reaction mixture was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) to give the desired compound of formulae 1-3 (2.65g, 60%) as a colorless oil.
(formula 1-4: 4- ((N- (3-fluorophenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3-fluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 1-3, 0.32g, 0.75mmol was dissolved in dimethylformamide (5mL), followed by the addition of potassium carbonate (0.31g, 2.24mmol) and morpholine (0.13mL, 1.49 mmol). The mixture was reacted at 60 ℃ for 1 day and diluted with saturated ammonium chloride solution. The organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) to give the desired compound of formulae 1-4 (0.13g, 45%).
(Compound 530: N- (3-fluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
A compound of formula 1-4 (methyl 4- ((N- (3-fluorophenyl) morpholine-4-carboxamido) methyl) benzoate; 0.108g, 0.290mmol) was dissolved in methanol (10mL), hydroxylamine (50.0 wt.% in water; 1.19mL, 19.4mmol) was added at room temperature, followed by potassium hydroxide (0.156g, 2.78mmol) and stirring at the same temperature for 16 hours. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The precipitated solid was filtered and dried to give the desired compound 530(0.062g, 57%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.14(brs，1H)，8.99(brs，1H)，7.65(d，2H，J＝7.0Hz)，7.38-7.30(m，3H)，7.05-6.85(m，3H)，4.89(s，1H)，3.44-3.42(m，4H)，3.18-3.15(m，4H)，2.08(s，3H)。MS(ESI)m/z 374(M++H)。
Example 136: synthesis of Compound 531
(formula 12-1: 4- ((4- (methoxycarbonyl) benzyl) (3- (trifluoromethyl) phenyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate, 1.040g, 2.192mmol, piperazine-1-carboxylic acid tert-butyl ester (0.817g, 4.385mmol) and potassium carbonate (0.909g, 6.577mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (10mL) at 60 ℃ and stirred at the same temperature for 1 day. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 60%) and concentrated to give the desired compound of formula 12-1 (0.679g, 59.4%) as a white solid.
(formula 12-2; 4- ((N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester hydrochloride)
A compound of formula 12-1 (4- ((4- (methoxycarbonyl) benzyl) (3- (trifluoromethyl) phenyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester; 0.360g, 0.690 mmo) at room temperaturel) dissolving in 1, 4-bisAlkane (5mL), then hydrochloric acid (4.0M 1, 4-bis)An alkane solution; 0.863mL, 3.451mmol) was added to the solution and stirred at the same temperature for 2 hours. The reaction mixture was then concentrated under reduced pressure. The obtained product was used without any purification process, and the compound of formula 12-2 (0.370g, 117.1%) was obtained as a pale yellow liquid.
(formula 12-3: 4- ((4- (2-hydroxy-2-methylpropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid ethyl ester)
A compound of formula 12-2 (methyl 4- ((N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate hydrochloride; 0.310g, 0.677mmol), 2, 2-dimethyloxirane (0.610mL, 6.770mmol) and potassium carbonate (0.936g, 6.770mmol) were mixed with ethanol (4 mL)/water (1mL) and heated under microwave irradiation at 110 ℃ for 1 hour, after which the temperature was lowered to room temperature. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 70%) and concentrated to give the desired compound of formula 12-3 (0.121g, 35.2%) as a colorless liquid.
(formula 12-4: 4- ((4- (2-fluoro-2-methylpropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid ethyl ester)
Ethyl 4- ((4- (2-hydroxy-2-methylpropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, a compound of formula 12-3 (0.121g, 0.238mmol) was dissolved in dichloromethane (10mL), followed by addition of DAST (0.034mL, 0.262mmol) at 0 ℃ and stirring at the same temperature for 2 hours. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 60%) and concentrated to give the desired compound of formula 12-4 (0.056g, 46.0%) as a colorless liquid.
(Compound 531: 4- (2-fluoro-2-methylpropyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
A compound of formula 12-4 (ethyl 4- ((4- (2-fluoro-2-methylpropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.056g, 0.110mmol), hydroxylamine (50.0 wt% aqueous solution; 0.067mL, 1.099mmol) and potassium hydroxide (0.031g, 0.550mmol) were dissolved in methanol (5mL) at room temperature and stirred at the same temperature for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was dehydrated with anhydrous magnesium sulfate and then concentrated under reduced pressure. The precipitated solid was filtered and dried to give the desired compound 531(0.045g, 82.5%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，9.20(brs，1H)，7.65(d，2H，J＝8.1Hz)，7.53-7.49(m，1H)，7.39-7.36(m，5H)，4.92(s，2H)，3.61-3.56(m，1H)，3.42-3.37(m，2H)，2.55-2.45(m，2H)，2.44-2.38(m，1H)，2.32-2.29(m，4H)，1.28(s，3H)，1.23(s，3H)；MS(ESI)m/z 497.2(M++H)。
Example 137: synthesis of Compound 532
(formula 6-2: 3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzonitrile)
The compound of formula 1-1 (3- (trifluoromethyl) aniline; 0.998mL, 8.068mmol) was dissolved in acetonitrile (60mL), and then 4- (bromomethyl) -3-fluorobenzonitrile (2.072g, 9.682mmol) and DIPEA (2.143mL, 12.102mmol) were added at room temperature and stirred at the same temperature for 1 day. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 5 to 20%) and concentrated to give the desired compound of formula 6-2 (2.380g, 64.4%) as a yellow liquid.
(formula 6-3: 3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid)
A compound of formula 6-2 (3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzonitrile; 2.310g, 7.850mmol) and lithium hydroxide (3.294g, 78.505mmol) were mixed with methanol (40mL)/H2O (20mL) was mixed and then stirred under reflux for 16 hours. Then, the temperature was lowered to room temperature, and the reaction mixture was concentrated under reduced pressure. 2M aqueous hydrochloric acid was added to adjust pH to 1, and then the precipitated solid was filtered and dried to obtain the desired compound of formula 6-3 (1.700g, 69.1%) as whiteIn the form of a colored solid.
(formula 6-4: 3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 6-3 (3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid; 1.700g, 5.427mmol), methanol (4.402mL, 108.540mmol), EDC (2.081g, 10.854mmol), HOBt (1.467g, 10.854mmol) and DIPEA (2.883mL, 16.281mmol) were dissolved in tetrahydrofuran (50mL) at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 10 to 40%) and concentrated to give the desired compound of formula 6-4 (1.500g, 84.5%) as a colorless liquid.
(formula 6-5: 3-fluoro-4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 6-4 (methyl 3-fluoro-4- (((3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 1.500g, 4.583mmol), 4-nitrophenyl chloroformate (1.848g, 9.167mmol) and potassium carbonate (1.900g, 13.750mmol) were dissolved in acetonitrile (80mL) at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 10 to 40%) and concentrated to give the desired compound of formula 6-5 (0.927g, 41.1%) as a colorless liquid.
(formula 6-6: 3-fluoro-4- ((N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl compound of formula 6-5 (3-fluoro-4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.129g, 0.262mmol), morpholine (0.046mL, 0.524mmol) and potassium carbonate (0.109g, 0.786mmol) were dissolved in N, N-dimethylformamide (5mL) at 60 ℃ and stirred at the same temperature for 2 days. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 60%) and concentrated to give the desired compound of formula 6-6 (0.094g, 81.5%) as a colorless liquid.
(Compound 532: N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
Methyl 3-fluoro-4- ((N- (3- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.094g, 0.213mmol) and hydroxylamine (50.0 wt.% in water; 0.071g, 2.134mmol) were dissolved in methanol (5mL), then potassium hydroxide (0.060g, 1.067mmol) was added at room temperature and stirred at the same temperature for 2 hours, and the reaction mixture was concentrated under reduced pressure. Diethyl ether (10mL) was added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 532(0.068g, 72.2%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ11.2(brs，1H)，9.13(brs，1H)，7.57-7.42(m，7H)，4.94(s，2H)，3.44-3.34(m，4H)，3.18-3.12(m，4H)；MS(ESI)m/z 442.1(M++H)。
Example 138: synthesis of Compound 533
(formula 7-4: 4- (((5-chloropyridin-2-yl) amino) methyl) benzoic acid methyl ester)
The compound of formula 7-3 (5-chloropyridin-2-amine, 2.000g, 15.557mmol) and methyl 4-formylbenzoate (2.554g, 15.557mmol) were dissolved in methanol (50mL), and then acetic acid (0.890mL, 15.557mmol) was added to the reaction solution and stirred at room temperature for 2 hours. Then, sodium cyanoborohydride (0.978g, 15.557mmol) was added and stirred for 1 day. Then, methanol was removed by air drying to precipitate a solid, and the resulting solid was filtered and dried to obtain the desired compound of formula 7-4 (2.700g, 62.7%) as a white solid.
(formula 7-5: 4- (((5-chloropyridin-2-yl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 7-4 (4- (((5-chloropyridin-2-yl) amino) methyl) benzoate; 1.000g, 3.614mmol) and 4-nitrophenyl chloroformate (0.801g, 3.975mmol) were dissolved in methylene chloride (30mL) at room temperature and stirred at the same temperature for 3 days, and the solid was filtered off. Then, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate: hexane ═ 20 to 30%) and concentrated to give the desired compound of formula 7-5 (0.160g, 10.0%) as a white solid.
(formula 7-6: 4- ((N- (5-chloropyridin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((5-chloropyridin-2-yl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, a compound of formula 7-5 (0.100g, 0.226mmol) and morpholine (0.024mL, 0.272mmol) were dissolved in dimethylformamide (10mL) at 60 ℃ and stirred at the same temperature for 12 hours. Then, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 30%) and concentrated to give the desired compound of formula 7-6 (0.032g, 36.3%) as a colorless oil.
(Compound 533: N- (5-Chloropyridin-2-yl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (5-chloropyridin-2-yl) morpholine-4-carboxamido) methyl) benzoate of the formula 7-6 (0.032g, 0.082mmol) was dissolved in methanol (10mL) at room temperature, and then hydroxylamine hydrochloride (0.028g, 0.410mmol), potassium hydroxide (0.046g, 0.821mmol) and hydroxylamine (50% by weight aqueous solution; 0.211mL, 1.642mmol) were added to the reaction solution and stirred at the same temperature for 6 hours. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. Dichloromethane and hexane were added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 533(0.011g, 34.3%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ8.31(d，1H，J＝1.5Hz)，7.79-7.76(m，1H)，7.64(d，2H，J＝7.3Hz)，7.29(d，2H，J＝7.8Hz)，7.08(d，1H，J＝8.9Hz)，4.95(s，2H)，3.46-3.45(m，4H)，3.24-3.23(m，4H)；MS(ESI)m/z391.1(M++H)。
Example 139: synthesis of Compound 543
(formula 1-4: 4- ((4- (2-morpholinoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
A compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.253g, 0.534mmol), 4- (2- (piperazin-1-yl) ethyl) morpholine (0.213g, 1.068mmol) and potassium carbonate (0.221g, 1.602mmol) were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 2 days. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formulae 1 to 4 (0.139g, 48.5%) as a colorless liquid.
(Compound 543, N- (4- (hydroxycarbamoyl) benzyl) -4- (2-morpholinoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4- (2-morpholinoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.139g, 0.260mmol and hydroxylamine (50% by weight in water; 0.159ml, 2.600mmol) of the compound of the formula 1-4 were dissolved in methanol (5ml) and potassium hydroxide (0.073g, 1.300mmol) was added. It was stirred at room temperature for 16 hours. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was dehydrated with anhydrous magnesium sulfate and then concentrated under reduced pressure. The precipitated solid was filtered and dried to give the desired compound 543(0.055g, 39.5%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.2(brs，1H)，9.10(brs，1H)，7.64-7.56(m，4H)，7.34(d，2H，J＝8.0Hz)，7.20(d，2H，J＝8.8Hz)，4.89(s，2H)，3.49-3.47(m，4H)，3.16-3.14(m，4H)，2.31-2.24(m，12H)；MS(ESI)m/z536.2(M++H)。
Example 140: synthesis of Compound 544
(formula 1-4: 4- ((4- (pyridin-4-yl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.230g, 0.484mmol), 1- (pyridin-4-yl) piperazine (0.159g, 0.968mmol) and potassium carbonate (0.201g, 1.451mmol) were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 2 days. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formulae 1 to 4 (0.104g, 43.0%) as a colorless liquid.
(Compound 544, N- (4- (hydroxycarbamoyl) benzyl) -4- (pyridin-4-yl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4- (pyridin-4-yl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.104g, 0.209mmol, and hydroxylamine (50 wt.% in water; 0.128ml, 2.086mmol) of the compound of formulae 1-4 were dissolved in methanol (5ml) and potassium hydroxide (0.059g, 1.043mmol) was added. It was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. Saturated aqueous sodium bicarbonate (30ml) was poured into the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 544(0.086g, 82.7%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，8.99(brs，1H)，8.10(d，2H，J＝6.4Hz)，7.63-7.58(m，4H)，7.35(d，2H，J＝8.4Hz)，7.29(d，2H，J＝8.0Hz)，6.72(d，2H，J＝6.4Hz)，4.93(s，2H)，3.28(s，4H)；MS(ESI)m/z500.2(M++H)。
Example 141: synthesis of Compound 545
(formula 1-4: 4- ((4- (2-morpholino-2-oxoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl ester of the compound of formula 1-3 (4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.213g, 0.450mmol), 1-morpholino-2- (piperazin-1-yl) ethanone (0.192g, 0.900mmol) and potassium carbonate (0.187g, 1.350mmol) were dissolved in N, N-dimethylformamide (5ml) and stirred at 60 ℃ for 2 days. Saturated aqueous sodium bicarbonate was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 1-4 (0.109g, 44.1%) as a colorless liquid.
(Compound 545, N- (4- (hydroxycarbamoyl) benzyl) -4- (2-morpholino-2-oxoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4- (2-morpholino-2-oxoethyl) -N- (4- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.109g, 0.199mmol and hydroxylamine (50% by weight in water; 0.122ml, 1.987mmol) of the compound of the formula 1-4 were dissolved in methanol (5ml) and potassium hydroxide (0.056g, 0.994mmol) was added. It was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. Saturated aqueous sodium bicarbonate (30ml) was poured into the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 545(0.057g, 52.0%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，8.99(brs，1H)，7.64-7.57(m，4H)，7.32-7.30(m，2H)，7.20(d，2H，J＝8.4Hz)，4.88(s，2H)，3.48-3.46(m，6H)，3.36-3.35(m，2H)，3.17(brs，4H)，3.08(s，2H)，2.26(brs，4H)；MS(ESI)m/z 550.2(M++H)。
Example 142: synthesis of Compound 577
(formula 6-2: 3-fluoro-4- ((4-trifluoromethyl) phenylamino) methyl) benzonitrile)
The compound of formula 6-1 (4- (bromomethyl) -3-fluorobenzonitrile; 1.10g, 5.06mmol) was dissolved in acetonitrile (60mL), followed by addition of 4- (trifluoromethyl) aniline (0.39mL, 4.30mmol) and N, N-diisopropylethylamine (1.14mL, 6.45mmol) at room temperature and stirring at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10-30%) and concentrated to give the desired compound of formula 6-2 (0.11g, 28%) as a colorless oil.
(formula 6-3: 3-fluoro-4- ((4- (trifluoromethyl) phenylamino) methyl) benzoic acid)
Reacting a compound of formula 6-2 (3-fluoro-4- ((4-trifluoromethyl) phenylamino) methyl) benzonitrile; 0.93g, 3.17mmol) and lithium hydroxide monohydrate (1.33g, 31.73mmol) were dissolved in methanol (30 mL)/water (15mL) and stirred under reflux for 16 h. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. A 2N aqueous hydrochloric acid solution was poured into the obtained concentrate, and the precipitated solid was filtered to obtain the desired compound of formula 6-3 (0.36g, 37%) as an off-white solid.
(formula 6-4: 3-fluoro-4- ((4- (trifluoromethyl) phenylamino) methyl) benzoic acid methyl ester)
A compound of formula 6-3 (3-fluoro-4- ((4- (trifluoromethyl) phenylamino) methyl) benzoic acid; 0.36g, 1.17mmol), methanol (0.95mL, 23.49mmol) and N, N-diisopropylethylamine (0.62mL, 3.52mmol) were dissolved in THF (30mL), then 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC, 0.45g, 2.35mmol) and 1-hydroxybenzotriazole hydrate (HOBt, 0.31g, 2.35mmol) were added at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10-40%) and concentrated to give the desired compound of formula 6-4 (0.11g, 28%) as a white solid.
(formula 6-5: 3-fluoro-4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 6-4 (methyl 3-fluoro-4- ((4- (trifluoromethyl) phenylamino) methyl) benzoate; 0.61g, 1.86mmol), 4-nitrophenyl chloroformate (0.75g, 3.72mmol) and potassium carbonate (0.77g, 5.59mmol) were dissolved in acetonitrile (50mL) at room temperature and stirred at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10-30%) and concentrated to give the desired compound of formula 6-5 (0.62g, 67%) as a colorless oil.
(formula 6-6: 3-fluoro-4- ((N- (4- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 6-5 (methyl 3-fluoro-4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.31g, 0.63mmol), morpholine (0.11mL, 1.25mmol) and potassium carbonate (0.26g, 1.88mmol) were dissolved in dimethylformamide (5mL) at 60 ℃ and stirred at the same temperature for 16 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 60%) and concentrated to give the desired compound of formula 6-6 (1.58g, 98%) as a colorless liquid.
(Compound 577: N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
Methyl 3-fluoro-4- ((N- (4- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.15g, 0.35mmol, of the compound of formula 6-6 was dissolved in MeOH (10mL), and hydroxylamine (50.0 wt.% in water; 0.21mL, 3.49mmol) and potassium hydroxide (0.078g, 1.40mmol) were added at room temperature and stirred at the same temperature overnight. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 577(0.084g, 54%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.2(brs，1H)，9.11(brs，1H)，7.63(d，2H，J＝8.8Hz)，7.52-7.43(m，3H)，7.28(d，2H，J＝8.4Hz)，4.92(s，2H)，3.41-3.39(m，4H)，3.16-3.15(m，4H)。
Example 143: synthesis of Compound 578
(formula 6-6: 4- ((3, 3-difluoro-N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamido) methyl) 3-fluorobenzoic acid methyl ester)
Methyl compound of formula 6-5 (3-fluoro-4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.31g, 0.63mmol), 3, 3-difluoroazetidine hydrochloride (0.16g, 1.25mmol) and potassium carbonate (0.26g, 1.88mmol) were dissolved in dimethylformamide (5mL) at 60 ℃ and stirred at the same temperature for 16 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10-40%) and concentrated to give the desired compound of formula 6-6 (0.19g, 68%) as a colorless liquid.
(Compound 578: 3, 3-difluoro-N- (2-fluoro-4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamide)
Methyl 4- ((3, 3-difluoro-N- (4- (trifluoromethyl) phenyl) azetidine-1-carboxamido) methyl) 3-fluorobenzoate of the formula 6-6 (0.19g, 0.42mmol) was dissolved in MeOH (10mL) and hydroxylamine (50.0 wt.% in water; 0.26mL, 4.27mmol) and potassium hydroxide (0.12g, 2.14mmol) were added at room temperature and stirred at the same temperature overnight. The reaction mixture was then concentrated under reduced pressure. Saturated aqueous sodium bicarbonate (50ml) was poured into the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 578(0.15g, 82%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.2(brs，1H)，9.10(brs，1H)，7.70(d，2H，J＝8.4Hz)，7.52-7.44(m，4H)，7.42-7.39(m，1H)，4.95(s，2H)，3.94-3.88(m，4H)。
Example 144: synthesis of Compound 580
(formula 1-4: 4- ((N- (4- (trifluoromethyl) phenyl) -1, 4-oxazepane-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl (4- (((4-nitrophenoxy) carbonyl) (4- (trifluoromethyl) phenyl) amino) methyl) benzoate, a compound of formula 1-3, 0.29g, 0.62mmol was dissolved in dimethylformamide, 1, 4-oxazepane (0.17g, 1.24mmol) and potassium carbonate (0.25g, 1.86mmol) were added, reacted at 60 ℃ for one day, then diluted with a saturated ammonium chloride solution, the organic layer was extracted with ethyl acetate, dehydrated with anhydrous sodium sulfate, filtered, then concentrated under reduced pressure, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 10-40%) and concentrated to give the desired compound of formula 1-4 (0.08g, 30%) as a colorless liquid.
(Compound 580, N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethyl) phenyl) -1, 4-oxazepane-4-carboxamide)
Methyl 4- ((N- (4- (trifluoromethyl) phenyl) -1, 4-oxaazepane-4-carboxamido) methyl) benzoate, 0.082g, 0.18mmol, of a compound of formulae 1-4 was dissolved in MeOH (10 ml). Hydroxylamine (50 wt% aqueous solution; 0.057ml, 0.939mmol) and potassium hydroxide (0.10g, 1.87mmol) were added and reacted at room temperature overnight. The concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 580(0.03g, 40%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.2(brs，1H)，9.19(brs，1H)，7.63-7.59(m，4H)，7.39(d，2H，J＝8.0Hz)，7.22(d，2H，J＝8.4Hz)，4.88(s，2H)，3.54-3.49(m，4H)，3.40-3.28(m，4H)，1.75-1.66(m，2H)。
Example 145: synthesis of Compound 651
(formula 12-5: 4- ((4- ((4-hydroxy-tetrahydro-2H-pyran-4-yl) methyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 12-2 (methyl 4- ((N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate hydrochloride; 0.15g, 0.35mmol), 1, 6-dioxaspiro [2, 5] octane (0.12mL, 1.06mmol) and N, N-diisopropylethylamine (0.30mL, 1.78mmol) were mixed with ethanol (10mL) and heated under microwave irradiation at 110 ℃ for 40 minutes, after which the temperature was lowered to room temperature. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 12-5 (0.11g, 58%) as a colorless liquid.
(Compound 651: 4- ((4-hydroxy-tetrahydro-2H-pyran-4-yl) methyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
A compound of formula 12-5 (methyl 4- ((4- ((4-hydroxy-tetrahydro-2H-pyran-4-yl) methyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.11g, 0.20mmol) and hydroxylamine (50.0 wt.% in water; 0.06mL, 1.04mmol) were dissolved in methanol (10mL), followed by addition of potassium hydroxide (0.11g, 2.09mmol) at room temperature and stirring at the same temperature for 16 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 651(0.07g, 70%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，9.02(brs，1H)，7.66(d，2H，J＝7.7Hz)，7.53-7.49(m，1H)，7.39-7.37(m，5H)，4.92(s，2H)，4.15(s，1H)，3.60-3.54(m，4H)，3.16(s，4H)，2.35(s，4H)，2.18(s，2H)，1.52-1.47(m，2H)，1.35-1.31(m，2H)。
Example 146: synthesis of Compound 683
(formula 1-4: 4- ((4-benzyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Reacting a compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenylamino) methyl) benzoate; 0.13g, 0.27mmol) was dissolved in dimethylformamide (2mL), then potassium carbonate (0.11g, 0.83mmol) and 1-benzylpiperazine (0.10mL, 0.55mmol) were added, the mixture was reacted at 40 ℃ for 16 hours, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 20-60%) and concentrated to give the desired compound of formulae 1-4 (0.07g, 53%) as a colorless oil.
(Compound 683: 4-benzyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4-benzyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.07g, 0.14mmol and hydroxylamine (50.0 wt.% aqueous solution; 0.045mL, 0.743mmol) of the compound of formula 1-4 were dissolved in methanol (10mL) at room temperature, followed by addition of potassium hydroxide (0.08g, 1.48mmol) and stirring at the same temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. Saturated sodium bicarbonate (50ml) was added to the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 683(0.06g, 89%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，9.03(brs，1H)，7.64(d，2H，J＝8.2Hz)，7.52-7.48(m，1H)，7.39-7.35(m，5H)，7.31-7.17(m，5H)，4.87(s，2H)，3.39(s，2H)，3.17(s，4H)，2.33(s，4H)。
Example 147: synthesis of Compound 684
(formula 1-4: 4- ((4- (3-methoxyphenyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoate)
The compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenylamino) methyl) benzoate; 0.13g, 0.28mmol) was dissolved in dimethylformamide (2mL), then potassium carbonate (0.11g, 0.85mmol) and 1- (3-methoxyphenyl) piperazine (0.10mL, 0.56mmol) were added, the mixture was reacted at 40 ℃ for 16 hours, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate, the organic layer was washed with saturated aqueous sodium chloride, dehydrated with anhydrous sodium sulfate, then concentrated under reduced pressure, the residue was purified by column chromatography (silica; ethyl acetate/hexane 10-40%) and concentrated to give the desired compound of formula 1-4 (0.07g, 47%), in the form of a colorless oil.
(Compound 684: N- (4- (hydroxycarbamoyl) benzyl) -4- (3-methoxyphenyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Methyl 4- ((4- (3-methoxyphenyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.07g, 0.13mmol) and hydroxylamine (50.0 wt% aqueous solution; 0.04mL, 0.67mmol) of the compound of formula 1-4 were dissolved in methanol (10mL) at room temperature, followed by addition of potassium hydroxide (0.07g, 1.34mmol) and stirring at the same temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. Saturated sodium bicarbonate (50ml) was added to the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 684(0.05g, 77%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝8.1Hz)，7.54-7.44(m，3H)，7.39-7.34(m，3H)，7.08(t，1H，J＝8.2Hz)，6.46-6.35(m，3H)，4.92(s，2H)，3.68(s，3H)，3.30(s，4H)，2.97(s，4H)。
Example 148: synthesis of Compound 716
(formula 12-6: 4- ((4-fluoro-tetrahydro-2H-pyran-4-yl) methyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((4- ((4-hydroxy-tetrahydro-2H-pyran-4-yl) methyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.28g, 0.52mmol) of the compound of formula 12-5 was dissolved in dichloromethane (15mL), followed by addition of DAST (0.08mL, 0.58mmol) at 0 ℃ and stirring at the same temperature for 3 hours. Then, a saturated aqueous sodium hydrogencarbonate solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 90%) and concentrated to give the desired compound of formula 12-6 (0.15g, 54%) as a colorless liquid.
(Compound 716: 4- ((4-Fluorotetrahydro-2H-pyran-4-yl) methyl) -N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
Reacting a compound of formula 12-6 (methyl 4- ((4-fluoro-tetrahydro-2H-pyran-4-yl) methyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.15g, 0.28mmol) and hydroxylamine (50.0 wt% aqueous solution; 0.08mL, 1.44mmol) was dissolved in MeOH (10mL), and potassium hydroxide (0.16g, 2.88mmol) was added to the mixture and stirred at room temperature and at the same temperature for 16 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound 716(0.06g, 41%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.1(brs，1H)，9.00(brs，1H)，7.65(d，2H，J＝7.7Hz)，7.53-7.49(m，1H)，7.39-7.35(m，5H)，4.92(s，2H)，3.64-3.61(m，2H)，3.52-3.49(m，2H)，3.22-3.16(m，4H)，2.46(s，2H)，2.33-2.29(m，4H)，1.69-1.67(m，4H)。
Example 149: synthesis of Compound 717
(formula 1-4: 4- ((4-phenyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Reacting a compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenylamino) methyl) benzoate; 0.12g, 0.26mmol) was dissolved in dimethylformamide (5mL), then potassium carbonate (0.10g, 0.79mmol) and 1-phenylpiperazine (0.08mL, 0.56mmol) were added, the mixture was reacted at 40 ℃ for 16 hours, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 5-70%) and concentrated to give the desired compound of formulae 1-4 (0.07g, 57%) as a colorless oil.
(Compound 717: N- (4- (hydroxycarbamoyl) benzyl) -4-phenyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
At room temperature, methyl 4- ((4-phenyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.07g, 0.15mmol) of the compound of formula 1-4 was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.04mL, 0.76mmol) and potassium hydroxide (0.08g, 1.52mmol) were added and stirred at the same temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. Saturated sodium bicarbonate (30ml) was added to the concentrate and stirred. The precipitated solid was filtered and dried to give the desired compound 717(0.05g, 70%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ7.65(d，2H，J＝8.2Hz)，7.58-7.46(m，3H)，7.39-7.31(m，3H)，7.20-7.16(m，2H)，6.87(d，2H，J＝8.0Hz)，6.80-6.76(m，1H)，4.95(s，2H)，3.32-3.30(m，4H)，2.98-2.95(m，4H)。
Example 150: synthesis of Compound 718
(formula 1-4: 4- ((4-benzhydryl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Reacting a compound of formula 1-3 (methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenylamino) methyl) benzoate; 0.14g, 0.29mmol) was dissolved in dimethylformamide (5mL), then potassium carbonate (0.12g, 0.88mmol) and 1-benzhydrylpiperazine (0.15g, 0.59mmol) were added, the mixture was reacted at 40 ℃ for 16 hours, dimethylformamide was removed under reduced pressure, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous sodium sulfate, the residue was purified by column chromatography (silica; ethyl acetate/hexane ═ 5-70%) and concentrated to give the desired compound of formulae 1-4 (0.13g, 77%) as a colorless oil.
(Compound 718: 4-Diphenylmethyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
At room temperature, methyl compound of formulae 1-4 (4- ((4-benzhydryl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.13g, 0.22mmol) was dissolved in methanol (10mL), and then hydroxylamine (50.0 wt% aqueous solution; 0.07mL, 1.14mmol) and potassium hydroxide (0.12g, 2.28mmol) were added and stirred at the same temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. The precipitated solid was filtered and dried to give the desired compound 718(0.06g, 46%) as a pale yellow solid.
1H NMR(400MHz，DMSO-d6)δ7.62-7.59(m，2H)，7.49-7.43(m，1H)，7.38-7.32(m，7H)，7.27-7.22(m，6H)，7.17-7.13(m，2H)，4.85(s，2H)，4.20(s，1H)，3.19(brs，4H)，2.13(brs，4H)。
Example 151: synthesis of Compound 765
(formula 10-8: 4- ((4-ethyl-N- (3- (hydroxymethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
A compound of formula 10-3 (methyl 4- (((3- (hydroxymethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.500g, 1.15mmol), 1-ethyl-piperazine (0.392g, 3.44mmol) and potassium carbonate (0.792g, 5.73mmol) were mixed with N, N-dimethylformaldehyde (5mL) at room temperature and stirred at the same temperature for 60 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium bicarbonate solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 10-8 (0.252g, 54%) as a pale yellow liquid.
(formula 10-9: 4- ((4-Ethyl-N- (3- (fluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 10-8 (methyl 4- ((4-ethyl-N- (3- (hydroxymethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.252g, 0.61mmol) was dissolved in dichloromethane (10mL) at room temperature, and diethylaminosulfur trifluoride (0.089mL, 0.67mmol) was added to the solution and stirred at the same temperature for 2 hours. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; methanol/dichloromethane ═ 10%) and concentrated to give the desired compound of formula 10-9 (0.210g, 83%) as a yellow liquid.
(Compound 765: 4-Ethyl-N- (3- (fluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
A compound of formula 10-9 (methyl 4- ((4-ethyl-N- (3- (fluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.210g, 0.508mmol) was dissolved in methanol (10mL) at room temperature, and then hydroxylamine (50 wt% aqueous solution; 3.11mL, 50.79mmol) and potassium hydroxide (0.285g, 5.08mmol) were added to the solution and stirred at the same temperature for 3 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained product was used without any purification process to obtain the desired compound 765(0.137g, 65.1%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ11.16(brs，1H)，9.10(brs，1H)，7.66(d，2H，J＝8.1Hz)，7.38-7.29(m，3H)，7.17(s，1H)，7.13-7.08(m，2H)，5.37(d，2H，J＝47.5Hz)，4.88(s，2H)，3.16(m，4H)，2.23(q，2H，J＝7.1Hz)，2.17(m，4H)，0.93(t，3H，J＝7.1Hz).MS(ESI)m/z 415(M++H)。
Example 152: synthesis of Compound 766
(formula 10-10: 4- ((3- (hydroxymethyl) phenyl) (4- (methoxycarbonyl) benzyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester)
A compound of formula 10-3 (methyl 4- (((3- (hydroxymethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 1.000g, 2.29mmol), 1-Boc-piperazine (1.280g, 6.87mmol) and potassium carbonate (1.583g, 11.46mmol) were mixed with N, N-dimethylformamide (7mL) at room temperature and stirred at the same temperature for 60 hours. Then, a saturated aqueous ammonium chloride solution was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium bicarbonate solution, dehydrated with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 60%) and concentrated to give the desired compound of formula 10-10 (0.263g, 24%) as a white solid.
(formula 10-11: 4- ((3- (fluoromethyl) phenyl) (4- (methoxycarbonyl) benzyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester)
At room temperature, the compound of formula 10-10 (4- ((3- (hydroxymethyl) phenyl) (4- (methoxycarbonyl) benzyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester; 0.263g, 0.54mmol) was dissolved in dichloromethane (10mL), and diethylaminosulfur trifluoride (0.079mL, 0.60mmol) was added to the solution and stirred at the same temperature for 2 hours. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 50%) and concentrated to give the desired compound of formula 10-11 (0.221g, 84%) as a colorless liquid.
(formula 10-12: 4- ((N- (3- (fluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester hydrochloride)
A compound of formula 10-11 (4- ((3- (fluoromethyl) phenyl) (4- (methoxycarbonyl) benzyl) carbamoyl) piperazine-1-carboxylic acid tert-butyl ester; 0.221g, 0.46mmol) was dissolved in 1, 4-bis (tert-butyl) acetate at room temperatureAlkane (5mL), then hydrochloric acid (4M1, 4-bis)An alkane solution; 5.689mL, 22.76mmol) was added to the solution and stirred at the same temperature for 1 hour. Then, the solvent was removed from the reaction mixture under reduced pressure, and ethyl acetate (10mL) and hexane (30mL) were added to the concentrate and stirred. Then, the precipitated solid was filtered, washed with hexane, and dried to obtain the desired compound of formula 10-12 (0.160g, 83%) as a white solid.
(formula 10-13: 4- ((N- (3- (fluoromethyl) phenyl) -4- (2-hydroxy-2-methylpropyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 10-12 (methyl 4- ((N- (3- (fluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate hydrochloride; 0.137g, 0.33mmol), 2, 2-dimethyloxirane (0.289mL, 3.25mmol) and potassium carbonate (0.224g, 1.62mmol) were mixed with ethanol (10mL) and heated under microwave irradiation at 110 ℃ for 20 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Water was poured into the obtained concentrate, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained product was used without any purification process to obtain the desired compound of formula 10-13 (0.129g, 87%) as a pale yellow liquid.
(formula 10-14: 4- ((4- (2-fluoro-2-methylpropyl) -N- (3- (fluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- ((N- (3- (fluoromethyl) phenyl) -4- (2-hydroxy-2-methylpropyl) piperazine-1-carboxamido) methyl) benzoate, a compound of formula 10-13 (0.129g, 0.28mmol) was dissolved in dichloromethane (10mL) at room temperature, and then diethylaminosulfur trifluoride (0.041mL, 0.31mmol) was added to the solution and stirred at the same temperature for 2 hours. Then, a saturated aqueous sodium bicarbonate solution was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 10 to 40%) and concentrated to give the desired compound of formula 10-14 (0.111g, 86%) as a colorless liquid.
(Compound 766: 4- (2-fluoro-2-methylpropyl) -N- (3- (fluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) piperazine-1-carboxamide)
Methyl 4- ((4- (2-fluoro-2-methylpropyl) -N- (3- (fluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, a compound of formula 10-14 (0.111g, 0.24mmol) was dissolved in methanol (10mL) at room temperature, and then hydroxylamine (50 wt% aqueous solution; 1.477mL, 24.16mmol) and potassium hydroxide (0.136g, 2.42mmol) were added to the solution and stirred at the same temperature for 3 hours. Then, a concentrate was obtained by removing the solvent from the reaction mixture under reduced pressure. Saturated aqueous sodium bicarbonate was poured into the obtained concentrate, and then the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The obtained product was used without any purification process to obtain the desired compound 766(0.087g, 78%) as a white solid.
1HNMR(400MHz，DMSO-d6)δ11.15(brs，1H)，9.04(brs，1H)，7.65(d，2H，J＝8.2Hz)，7.37-7.32(m，3H)，7.16(s，1H)，7.12-7.08(m，2H)，5.37(d，2H，J＝47.5Hz)，4.87(s，2H)，3.16(m，4H)，2.36(s，2H，J＝23.0Hz)，2.33(m，4H)，1.29(s，3H)，1.23(s，3H)。MS(ESI)m/z 461(M++H)。
Example 153: synthetic Compound 771
(formula 1-2: 4- (((2- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-1 (2- (trifluoromethoxy) aniline; 0.385mL, 4.625mmol), methyl 4- (bromomethyl) benzoate (1.271g, 5.550mmol) and DIPEA (1.228mL, 6.937mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (0.806g, 53.6%) as a colorless liquid.
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (2- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((2- (trifluoromethoxy) phenyl) amino) methyl) benzoate; 0.806g, 2.477mmol), 4-nitrophenyl chloroformate (0.549g, 2.725mmol) and potassium carbonate (0.685g, 4.955mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-3 (1.172g, 96.5%) as a yellow liquid.
(formula 1-4: 4- ((N- (2- (trifluoromethoxy) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- (((((4-nitrophenoxy) carbonyl) (2- (trifluoromethoxy) phenyl) amino) methyl) benzoate, 1.172g, 2.390mmol), morpholine (2.082g, 23.899mmol) and potassium carbonate (1.652g, 11.950mmol) were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 h. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.948g, 90.4%) as a colorless liquid.
(Compound 771: N- (4- (hydroxycarbamoyl) benzyl) -N- (2- (trifluoromethoxy) phenyl) morpholine-4-carboxamide)
A compound of formula 1-4 (methyl 4- ((N- (2- (trifluoromethoxy) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.948g, 2.162mmol), hydroxylamine (50.0 wt.% in water; 1.323mL, 21.624mmol) and potassium hydroxide (1.213g, 21.624mmol) were dissolved in methanol (3mL) at room temperature and stirred at the same temperature for 3 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 80%) and concentrated to give the desired compound 771(0.656g, 69.0%) as a white solid.
1H NMR(400MHz，DMSO-d6)d 7.68(d，2H，J＝8.0Hz)，7.39～7.32(m，6H)，4.76(s，2H)，3.36～3.35(m，4H)，3.10(m，4H)；MS(ESI)m/z440.1(M++H)。
Example 154: synthesis of Compound 772
(formula 1-2: 4- (((4- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (4- (trifluoromethoxy) aniline; 0.385mL, 4.625mmol), methyl 4- (bromomethyl) benzoate (1.271g, 5.550mmol) and DIPEA (1.228mL, 6.937mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (0.645g, 42.8%) as a colorless liquid.
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((4- (trifluoromethoxy) phenyl) amino) methyl) benzoate; 0.645g, 1.981mmol), 4-nitrophenyl chloroformate (0.439g, 2.180mmol) and potassium carbonate (0.548g, 3.963mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-3 (0.633g, 65.1%) as a white solid.
(formula 1-4: 4- ((N- (4- (trifluoromethoxy) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (4- (trifluoromethoxy) phenyl) amino) methyl) benzoate, 0.633g, 1.291mmol), morpholine (1.125g, 12.909mmol) and potassium carbonate (0.892g, 6.454mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 h. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.555g, 98.1%) as a colorless liquid.
(Compound 772: N- (4- (hydroxycarbamoyl) benzyl) -N- (4- (trifluoromethoxy) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (4- (trifluoromethoxy) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.555g, 1.266mmol, hydroxylamine (50.0 wt.% in water; 0.774mL, 12.660mmol) and potassium hydroxide (0.710g, 12.660mmol) of the compounds of formulae 1-4 were dissolved in methanol (3mL) at room temperature and stirred at the same temperature for 3 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50-80%) and concentrated to give the desired compound 772(0.544g, 97.8%) as a white solid.
1H NMR(400MHz，DMSO-d6)d 10.33(brs，1H)，9.78(brs，1H)，7.65(d，2H，J＝7.8Hz)，7.42(t，1H，J＝8.2Hz)，7.37(d，2H，J＝7.8Hz)，7.18(d，1H，J＝7.6Hz)，7.12(s，1H)，7.03(d，1H，J＝8.3Hz)，4.91(s，2H)，3.43(m，4H)，3.17(m，4H)；MS(ESI)m/z 440.1(M++H)。
Example 155: synthesis of Compound 773
(formula 1-2: 4- (((3- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-1 (3- (trifluoromethoxy) aniline; 0.385mL, 4.625mmol), methyl 4- (bromomethyl) benzoate (1.271g, 5.550mmol) and DIPEA (1.228mL, 6.937mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (0.607g, 40.3%) as a white solid.
(formula 1-3: 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethoxy) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((3- (trifluoromethoxy) phenyl) amino) methyl) benzoate; 0.607g, 1.865mmol), 4-nitrophenylchloroformate (0.413g, 2.051mmol) and potassium carbonate (0.515g, 3.730mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-3 (0.649g, 71.0%) as a white solid.
(formula 1-4: 4- ((N- (3- (trifluoromethoxy) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethoxy) phenyl) amino) methyl) benzoate, a compound of formula 1-3 (0.649g, 1.324mmol), morpholine (1.154g, 13.243mmol) and potassium carbonate (0.915g, 6.621mmol) were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 h. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.512g, 88.2%) as a colorless liquid.
(Compound 773: N- (4- (hydroxycarbamoyl) benzyl) -N- (3- (trifluoromethoxy) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (3- (trifluoromethoxy) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.512g, 1.168mmol, hydroxylamine (50.0 wt.% in water; 0.714mL, 11.679mmol) and potassium hydroxide (0.655g, 11.679mmol) of the compounds of formulae 1-4 were dissolved in methanol (3mL) at room temperature and stirred at the same temperature for 3 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 80%) and concentrated to give the desired compound 773(0.417g, 81.3%) as a white solid.
1H NMR(400MHz，DMSO-d6)d 10.35(brs，1H)，8.68(brs，1H)，7.67(d，2H，J＝8.0Hz)，7.37(d，2H，J＝8.0Hz)，7.31～7.24(m，4H)，4.88(s，2H)，3.42(m，4H)，3.15(m，4H)；MS(ESI)m/z 440.1(M++H)。
Example 156: synthesis of Compound 774
(formula 1-2: 4- (((2-methoxy-5- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-1 (2-methoxy-5- (trifluoromethyl) aniline; 0.385mL, 4.625mmol), methyl 4- (bromomethyl) benzoate (1.271g, 5.550mmol) and DIPEA (1.228mL, 6.937mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (0.607g, 40.3%) as a white solid.
(formula 1-3: 4- (((2-methoxy-5- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((2-methoxy-5- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.607g, 1.865mmol), 4-nitrophenylchloroformate (0.413g, 2.051mmol) and potassium carbonate (0.515g, 3.730mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-3 (0.649g, 71.0%) as a white solid.
(formula 1-4: 4- ((N- (2-methoxy-5- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 1-3 (methyl 4- (((2-methoxy-5- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.649g, 1.324mmol), morpholine (1.154g, 13.243mmol) and potassium carbonate (0.915g, 6.621mmol) were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 hours. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.512g, 88.2%) as a colorless liquid.
(Compound 774: N- (4- (hydroxycarbamoyl) benzyl) -N- (2-methoxy-5- (trifluoromethyl) phenyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2-methoxy-5- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.512g, 1.168mmol, hydroxylamine (50.0 wt.% in water; 0.714mL, 11.679mmol) and potassium hydroxide (0.655g, 11.679mmol) were dissolved in methanol (3mL) at room temperature and stirred at the same temperature for 3 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50-80%) and concentrated to give the desired compound 774(0.417g, 81.3%) as a white solid.
1H NMR(400MHz，DMSO-d6)d 10.35(brs，1H)，8.68(brs，1H)，7.67(d，2H，J＝8.0Hz)，7.37(d，2H，J＝8.0Hz)，7.31～7.24(m，4H)，4.88(s，2H)，3.42(m，4H)，3.15(m，4H)；MS(ESI)m/z 454.1(M++H)。
Example 157: synthesis of Compound 776
(formula 1-2: 4- (((2-chloro-5- (trifluoromethyl) phenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (2-chloro-5- (trifluoromethyl) aniline; 0.352mL, 2.557mmol), methyl 4- (bromomethyl) benzoate (0.703g, 3.068mmol) and DIPEA (0.679mL, 3.835mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 1-2 (0.752g, 85.6%) as a white solid.
(formula 1-3: 4- (((2-chloro-5- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((2-chloro-5- (trifluoromethyl) phenyl) amino) methyl) benzoate; 0.752g, 2.188mmol), 4-nitrophenyl chloroformate (0.485g, 2.407mmol) and potassium carbonate (0.605g, 4.376mmol) were dissolved in acetonitrile (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-3 (0.903g, 81.1%) as a white solid.
(formula 1-4: 4- ((N- (2-chloro-5- (trifluoromethyl) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
A compound of formula 1-3 (methyl 4- (((2-chloro-5- (trifluoromethyl) phenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate; 0.903g, 1.775mmol), morpholine (1.546g, 17.747mmol) and potassium carbonate (1.226g, 8.873mmol) were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 hours. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30%) and concentrated to give the desired compound of formulae 1 to 4 (0.582g, 71.8%) as a colorless liquid.
(Compound 776: N- (2-chloro-5- (trifluoromethyl) phenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2-chloro-5- (trifluoromethyl) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.582g, 1.274mmol, hydroxylamine (50.0 wt.% in water; 0.779mL, 12.740mmol) and potassium hydroxide (0.715g, 12.740mmol) of the compounds of formulae 1-4 were dissolved in methanol (3mL) at room temperature and stirred at the same temperature for 3 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 80%) and concentrated to give the desired compound 776(0.521g, 89.3%) as a milky white solid.
1H NMR(400MHz，DMSO-d6)d 10.33(brs，1H)，8.67(brs，1H)，7.76(d，1H，J＝8.1Hz)，7.65(s，1H)，7.64(s，2H)，7.60(d，1H，J＝8.3Hz)，7.36(d，2H，J＝8.1Hz)，4.82(s，2H)，3.33(m，4H)，3.13～3.12(m，4H)；MS(ESI)m/z 458.0(M++H)。
Example 158: synthesis of Compound 778
(formula 7-4: 4- (((6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoic acid methyl ester)
At room temperature, the compound of formula 7-3 (6- (trifluoromethyl) pyridin-2-amine (0.500g, 3.084mmol), methyl 4- (bromomethyl) benzoate (1.413g, 6.169mmol) and DIPEA (1.092mL, 6.169mmol) were dissolved in acetonitrile (10mL) and stirred at the same temperature for 16 hours, then, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate.
(formula 7-5: 4- ((((4-Nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoic acid methyl ester)
A compound of formula 7-4 (methyl 4- (((6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.588g, 1.895mmol), 4-nitrophenyl chloroformate (0.420g, 2.085mmol) and potassium carbonate (0.524g, 3.790mmol) were dissolved in acetonitrile (10mL) at room temperature and stirred at the same temperature for 16 hours. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formula 7-5 (0.295g, 32.7%) as a yellow liquid.
(formula 7-6: 4- ((N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.295g, 0.621mmol), morpholine (0.541mL, 6.206mmol) and potassium carbonate (0.429g, 3.103mmol) were dissolved in N, N-dimethylformamide (5mL) at room temperature and stirred at the same temperature for 1 hour. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 7 to 6 (0.112g, 42.6%) as a colorless liquid.
(Compound 778: N- (4- (hydroxycarbamoyl) benzyl) -N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamide)
Methyl 4- ((N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoate, 0.112g, 0.265mmol, hydroxylamine (50.0 wt.% in water; 0.162mL, 2.645mmol) and potassium hydroxide (0.148g, 2.645mmol) of the compound of formula 7-6 were dissolved in methanol (5mL) at room temperature and stirred at the same temperature for 1 hour. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50 to 80%) and concentrated to give the desired compound 778(0.072g, 64.1%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.88(t，1H，J＝7.9Hz)，7.70(d，2H，J＝8.1Hz)，7.53(d，2H，J＝8.1Hz)，7.35(d，1H，J＝7.4Hz)，7.24(d，1H，J＝8.4Hz)，5.11(s，2H)，3.55(t，4H，J＝4.7Hz)，3.36(t，4H，J＝4.6Hz)；MS(ESI)m/z 425.1(M++H)。
Example 159: synthesis of Compound 791
(formula 1-2: 4- (((2, 3-difluorophenyl) amino) methyl) benzoic acid methyl ester)
The compound of formula 1-1 (2, 3-difluoroaniline (0.300g, 2.324mmol), methyl 4- (bromomethyl) benzoate (0.639g, 2.788mmol) and DIPEA (0.809mL, 4.647mmol) were dissolved in acetonitrile (3mL) at room temperature and stirred at the same temperature for 17 hours, water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure, the concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 10%) and concentrated to give the desired compound of formula 1-2 (0.284g, 44.1%) as a white solid.
(formula 1-3: 4- (((2, 3-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoic acid methyl ester)
A compound of formula 1-2 (methyl 4- (((2, 3-difluorophenyl) amino) methyl) benzoate; 0.275g, 0.992mmol), 4-nitrophenyl chloroformate (0.240g, 1.190mmol) and potassium carbonate (0.274g, 1.948mmol) were dissolved in dichloromethane (3mL) at room temperature and stirred at the same temperature for 20 hours. Water was poured into the reaction mixture, and the organic layer was extracted with dichloromethane. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1-3 (0.177g, 40.3%) as a colorless oil.
(formula 1-4: 4- ((N- (2, 3-difluorophenyl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((2, 3-difluorophenyl) ((4-nitrophenoxy) carbonyl) amino) methyl) benzoate, morpholine (0.338mL, 3.843mmol) and potassium carbonate (0.265g, 1.921mmol) were dissolved in N, N-dimethylformamide (2mL) at 50 ℃ and stirred at the same temperature for 18 h. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 1 to 4 (0.113g, 75.3%) as a colorless liquid.
(Compound 791: N- (2, 3-difluorophenyl) -N- (4- (hydroxycarbamoyl) benzyl) morpholine-4-carboxamide)
Methyl 4- ((N- (2, 3-difluorophenyl) morpholine-4-carboxamido) methyl) benzoate, 0.105g, 0.269mmol, hydroxylamine (50.0 wt.% in water; 0.329mL, 5.379mmol) and potassium hydroxide (0.151g, 2.690mmol) of the compounds of formulae 1-4 were mixed and stirred at room temperature for 1 hour. A saturated aqueous sodium bicarbonate solution was poured into the reaction solution, and the organic layer was extracted with dichloromethane. The organic layer was dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure to give the desired compound 791(0.074g, 70.3%) as a pale brown solid.
MS(ESI)m/z 392.13(M++H)。
Example 160: synthesis of Compound 797
(formula 12-7: 4- (((3S, 5R) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethyl) phenyl) amino) methyl) benzoate, 1.263g, 2.662mmol), (2S, 6R) -2, 6-dimethylpiperazine (1.520g, 13.309mmol) and potassium carbonate (0.736g, 5.323mmol) of the compound of formula 1-3 were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 hours. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20 to 50%) and concentrated to give the desired compound of formulae 12 to 7 (0.726g, 60.7%) as a yellow liquid.
(formula 12-8: 4- (((3S, 5R) -4-ethyl-3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 12-7 (methyl 4- (((3S, 5R) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.200g, 0.445mmol), iodoethane (0.107mL, 1.335mmol) and potassium carbonate (0.184g, 1.335mmol) were mixed with acetonitrile (10mL) and heated under microwave irradiation at 120 ℃ for 20 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 30 to 80%) and concentrated to give the desired compound of formulae 12 to 8 (0.109g, 51.3%) as a colorless liquid.
(Compound 797 (3S, 5R) -4-Ethyl-N- (4- (hydroxycarbamoyl) benzyl) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
A compound of formula 12-8 (methyl 4- (((3S, 5R) -4-ethyl-3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.109g, 0.228mmol), hydroxylamine (50.0 wt.% in water; 0.140mL, 2.283mmol) and potassium hydroxide (0.128g, 2.283mmol) were dissolved in methanol (2mL) at room temperature and stirred at the same temperature for 1 hour. Then, the solvent was removed from the reaction mixture under reduced pressure, and a saturated aqueous sodium bicarbonate solution (20mL) was added to the concentrate and stirred. The precipitated solid was then filtered, washed with diethyl ether, and dried to give the desired compound 797(0.071g, 65.0%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.68(d，2H，J＝8.3Hz)，7.55(t，1H，J＝8.2Hz)，7.45～7.42(m，5H)，4.98(s，2H)，3.65(d，2H，J＝11.6Hz)，2.89(q，2H，J＝7.1Hz)，2.52～2.42(m，4H)，0.96(d，6H，J＝5.8Hz)，0.97(s，3H)，0.95(s，3H)，0.86(t，3H，J＝7.2Hz)；MS(ESI)m/z479.1(M++H)。
Example 161: synthesis of Compound 800
(formula 1-4: 4- ((2, 6-dimethyl-N- (3- (trifluoromethoxy) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl (4- ((((4-nitrophenoxy) carbonyl) (3- (trifluoromethoxy) phenyl) amino) methyl) benzoate, 0.200g, 0.408mmol), 2, 6-dimethylmorpholine (0.235g, 2.039mmol) and potassium carbonate (0.169g, 1.224mmol) were dissolved in N, N-dimethylformamide (5mL) at 40 ℃ and stirred at the same temperature for 16 hours. Then, the temperature was lowered to room temperature, and the reaction was completed. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 1 to 4 (0.166g, 87.3%) as a yellow liquid.
(Compound 800: N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (3- (trifluoromethoxy) phenyl) morpholine-4-carboxamide)
Methyl 4- ((2, 6-dimethyl-N- (3- (trifluoromethoxy) phenyl) morpholine-4-carboxamido) methyl) benzoate, 0.166g, 0.356mmol, hydroxylamine (50.0 wt.% in water; 0.218mL, 3.559mmol) and potassium hydroxide (0.200g, 3.559mmol) were dissolved in methanol (2mL) at room temperature and stirred at the same temperature for 3 hours. The precipitated solid was then filtered, washed with hexane, and dried to give the desired compound 800(0.098g, 59.0%) as a pale cream solid.
1H NMR(400MHz，CD3OD)d 7.68(d，2H，J＝8.4Hz)，7.43(d，2H，J＝8.4Hz)，7.43(t，1H，J＝8.1Hz)，7.16(ddd，1H，J＝8.2，2.0，0.8Hz)，7.06(d，1H，J＝0.9Hz)，7.04(d，1H，J＝1.0Hz)，4.96(s，2H)，3.66(d，2H，J＝12.4Hz)，3.44～3.40(m，2H)，2.38(dd，2H，J＝13.2，10.6Hz)，1.04(s，3H)，1.02(s，3H)；MS(ESI)m/z 468.1(M++H)。
Example 162: synthesis of Compound 801
(formula 1-4: 4- ((2, 6-dimethyl-N- (2- (trifluoromethoxy) phenyl) morpholine-4-formamido) methyl) benzoic acid methyl ester)
Methyl 4- ((((4-nitrophenoxy) carbonyl) (2- (trifluoromethoxy) phenyl) amino) methyl) benzoate, 2.300 g, 0.612mmol, 2, 6-dimethylmorpholine (0.352g, 3.059mmol) and potassium carbonate (0.254g, 1.835mmol) were dissolved in N, N-dimethylformamide (5mL) at room temperature and stirred at the same temperature for 16 hours. Water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 1 to 4 (0.116g, 40.7%) as a yellow liquid.
(Compound 801: N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (2- (trifluoromethoxy) phenyl) morpholine-4-carboxamide)
A compound of formulae 1-4 (methyl 4- ((2, 6-dimethyl-N- (2- (trifluoromethoxy) phenyl) morpholine-4-carboxamido) methyl) benzoate; 0.116g, 0.249mmol), hydroxylamine (50.0 wt.% in water; 0.152mL, 2.487mmol) and potassium hydroxide (0.140g, 2.487mmol) were dissolved in methanol (2mL) at room temperature and stirred at the same temperature for 3 hours. Then, water was poured into the reaction mixture and the organic layer was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (Waters, C18; acetonitrile/0.1% aqueous trifluoroacetic acid 5-70%) and concentrated on a SPE cartridge to give the desired compound 801 (0.020g, 17.5%) as a white solid.
1H NMR(400MHz，DMSO-d6)d 7.66(d，2H，J＝8.4Hz)，7.40～7.32(m，6H)，4.73(s，2H)，3.52(s，2H)，3.25～3.18(m，2H)，2.25(dd，2H，J＝13.0，10.7Hz)，0.92(s，3H)，0.91(s，3H)；MS(ESI)m/z 468.1(M++H)。
Example 163: synthesis of Compound 802
(formula 12-8: 4- (((3S, 5R) -4-benzyl-3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
Methyl 4- (((3S, 5R) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate, 0.200g, 0.445mmol, (bromomethyl) benzene (2.324g, 1.335mmol) and potassium carbonate (0.184g, 1.335mmol) of the compound of formula 12-7 were mixed with acetonitrile (3mL) and heated under microwave irradiation at 120 ℃ for 40 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 12-8 (0.113g, 47.1%) as a colorless liquid.
(Compound 802 (3S, 5R) -4-benzyl-N- (4- (hydroxycarbamoyl) benzyl) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
A compound of formula 12-8 (methyl 4- (((3S, 5R) -4-benzyl-3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.113g, 0.209mmol), hydroxylamine (50.0 wt% in water; 0.128mL, 2.094mmol) and potassium hydroxide (0.118g, 2.094mmol) were dissolved in methanol (2mL) at room temperature and stirred at the same temperature for 3 hours. Then, the solvent was removed from the reaction mixture under reduced pressure, and a saturated aqueous sodium bicarbonate solution (10mL) was added to the concentrate and stirred. The precipitated solid was then filtered, washed with hexane, and dried to give the desired compound 802(0.010g, 8.8%) as a cream solid.
1H NMR(400MHz，DMSO-d6)d 7.63(d，2H，J＝8.3Hz)，7.50(t，1H，J＝8.0Hz)，7.39～7.38(m，3H)，7.31～7.25(m，6H)，7.18～7.15(m，1H)，4.88(s，2H)，3.65(s，2H)，3.57(s，2H)，3.52(s，2H)，2.38～2.29(m，2H)，0.81(s，3H)，0.79(s，3H)；MS(ESI)m/z 541.1(M++H)。
Example 164: synthesis of Compound 803
(formula 12-8: 4- (((3S, 5R) -3, 5-dimethyl-4- (2, 2, 3, 3, 3-pentafluoropropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 12-7 (methyl 4- (((3S, 5R) -3, 5-dimethyl-N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.200g, 0.445mmol), 2, 2, 3, 3, 3-pentafluoropropyltrifluoromethanesulfonate (0.221mL, 1.335mmol) and potassium carbonate (0.184g, 1.335mmol) were mixed with acetonitrile (3mL) and heated under microwave irradiation at 120 ℃ for 30 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 20%) and concentrated to give the desired compound of formulae 12-8 (0.042g, 16.2%) as a colorless liquid.
(Compound 803 (3S, 5R) -N- (4- (hydroxycarbamoyl) benzyl) -3, 5-dimethyl-4- (2, 2, 3, 3, 3-pentafluoropropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamide)
A compound of formula 12-8 (methyl 4- (((3S, 5R) -3, 5-dimethyl-4- (2, 2, 3, 3, 3-pentafluoropropyl) -N- (3- (trifluoromethyl) phenyl) piperazine-1-carboxamido) methyl) benzoate; 0.042g, 0.072mmol), hydroxylamine (50.0 wt% in water; 0.044mL, 0.722mmol) and potassium hydroxide (0.041g, 0.722mmol) were dissolved in methanol (2mL) at room temperature and stirred at the same temperature for 1 hour. Then, the solvent was removed from the reaction mixture under reduced pressure, and a saturated aqueous sodium bicarbonate solution (10mL) was added to the concentrate and stirred. Then, the precipitated solid was filtered, washed with hexane, and dried to obtain the desired compound 803(0.016g, 38.0%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.69(d，2H，J＝8.3Hz)，7.53(t，1H，J＝7.8Hz)，7.43～7.38(m，5H)，4.96(s，2H)，3.59(d，2H，J＝12.5Hz)，2.63～2.53(m，4H)，1.36～1.31(m，2H)，0.97(s，3H)，0.96(s，3H)；MS(ESI)m/z583.2(M++H)。
Example 165: synthesis of Compound 826
(formula 7-6: 4- ((4-Ethyl-N- (6- (trifluoromethyl) pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.500g, 1.052mmol), 1-ethylpiperazine (0.240g, 2.104mmol) and potassium carbonate (0.291g, 2.104mmol) were mixed with N, N-dimethylformamide (3mL) and heated under microwave irradiation at 150 ℃ for 30 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 7 to 6 (0.276g, 58.2%) as a yellow liquid.
(Compound 826: 4-Ethyl-N- (4- (hydroxycarbamoyl) benzyl) -N- (6- (trifluoromethyl) pyridin-2-yl) piperazine-1-carboxamide)
A compound of formula 7-6 (methyl 4- ((4-ethyl-N- (6- (trifluoromethyl) pyridin-2-yl) piperazine-1-carboxamido) methyl) benzoate; 0.276g, 0.613mmol), hydroxylamine (50.0 wt% aqueous solution; 0.375mL, 6.127mmol) and potassium hydroxide (0.344g, 6.127mmol) were dissolved in methanol (1mL) at room temperature and stirred at the same temperature for 1 hour. The solvent was removed from the reaction mixture, and aqueous sodium bicarbonate solution (20mL) was added to the concentrate and stirred. The precipitated solid was then filtered and dried to give the desired compound 826(0.089g, 32.2%) as a yellow solid.
1H NMR(400MHz，CD3OD)d 7.88(t，1H，J＝7.9Hz)，7.71(d，2H，J＝8.3Hz)，7.49(d，2H，J＝8.3Hz)，7.35(d，1H，J＝7.5Hz)，7.20(d，1H，J＝8.5Hz)，5.10(s，2H)，3.40(t，4H，J＝4.9Hz)，2.40(q，2H，J＝7.3Hz)，2.35(t，4H，J＝5.0Hz)，1.07(t，3H，J＝7.2Hz)；MS(ESI)m/z452.1(M++H)。
Example 166: synthesis of Compound 827
(formula 7-6: 4- ((2, 6-dimethyl-N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.100g, 0.210mmol), 2, 6-dimethylmorpholine (0.048g, 0.421mmol) and potassium carbonate (0.058g, 0.421mmol) were mixed with N, N-dimethylformamide (3mL) and heated under microwave irradiation at 150 ℃ for 30 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 7-6 (0.076g, 79.7%) as a yellow liquid.
(Compound 827: N- (4- (hydroxycarbamoyl) benzyl) -2, 6-dimethyl-N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamide)
A compound of formula 7-6 (methyl 4- ((2, 6-dimethyl-N- (6- (trifluoromethyl) pyridin-2-yl) morpholine-4-carboxamido) methyl) benzoate; 0.076g, 0.168mmol), hydroxylamine (50.0 wt.% in water; 0.103mL, 1.684mmol) and potassium hydroxide (0.094g, 1.684mmol) were dissolved in methanol (1mL) at room temperature and stirred at the same temperature for 1 hour. The solvent was removed from the reaction mixture, and aqueous sodium bicarbonate solution (20mL) was added to the concentrate and stirred. Then, the precipitated solid was filtered and dried to obtain the desired compound 827(0.042g, 55.5%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.88(t，1H，J＝7.7Hz)，7.70(d，2H，J＝8.4Hz)，7.53(d，2H，J＝8.4Hz)，7.36(d，1H，J＝7.4Hz)，7.21(d，1H，J＝8.5Hz)，5.12(s，2H)，3.71(d，2H，J＝12.5Hz)，3.45～3.38(m，2H)，2.53(dd，2H，J＝10.7，13.1Hz)，1.07(s，3H)，1.05(s，3H)；MS(ESI)m/z 453.1(M++H)
Example 167: synthesis of Compound 828
(formula 7-6: 4- ((N- (6- (trifluoromethyl) pyridin-2-yl) piperidine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.100g, 0.210mmol), piperidine (0.036g, 0.421mmol) and potassium carbonate (0.058g, 0.421mmol) were mixed with N, N-dimethylformamide (3mL) and heated under microwave irradiation at 150 ℃ for 30 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formulae 7 to 6 (0.063g, 71.1%) as a yellow liquid.
(Compound 828: N- (4- (hydroxycarbamoyl) benzyl) -N- (6- (trifluoromethyl) pyridin-2-yl) piperidine-1-carboxamide)
Methyl 4- ((N- (6- (trifluoromethyl) pyridin-2-yl) piperidine-1-carboxamido) methyl) benzoate, 0.063g, 0.149mmol, hydroxylamine (50.0 wt.% in water; 0.091mL, 1.495mmol) and potassium hydroxide (0.084g, 1.495mmol) of the compound of formula 7-6 were dissolved in methanol (1mL) at room temperature and stirred at the same temperature for 1 hour. The solvent was removed from the reaction mixture, and aqueous sodium bicarbonate solution (20mL) was added to the concentrate and stirred. The precipitated solid was then filtered, washed with hexane and dried to give the desired compound 828(0.017g, 26.6%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.86(t，1H，J＝8.0Hz)，7.70(d，2H，J＝8.3Hz)，7.54(d，2H，J＝8.4Hz)，7.33(d，1H，J＝7.4Hz)，7.16(d，1H，J＝8.6Hz)，5.11(s，2H)，3.50～3.39(m，4H)，1.65～1.53(m，6H)；MS(ESI)m/z 423.1(M++H)。
Example 168: synthesis of Compound 829
(formula 7-6 (S) -4- ((3-fluoro-N- (6- (trifluoromethyl) pyridin-2-yl) pyrrolidine-1-carboxamido) methyl) benzoic acid methyl ester)
A compound of formula 7-5 (methyl 4- ((((4-nitrophenoxy) carbonyl) (6- (trifluoromethyl) pyridin-2-yl) amino) methyl) benzoate; 0.100g, 0.210mmol), (S) -3-fluoropyrrolidine (0.037g, 0.421mmol) and potassium carbonate (0.058g, 0.421mmol) were mixed with N, N-dimethylformamide (3mL) and heated under microwave irradiation at 150 ℃ for 30 minutes. Then, the temperature was lowered to room temperature, and the reaction was completed. Then, water was poured into the reaction mixture, and the organic layer was extracted with ethyl acetate. Then, the organic layer was washed with a saturated aqueous sodium chloride solution, dehydrated with anhydrous magnesium sulfate, filtered, and then concentrated under reduced pressure. The concentrate was purified by column chromatography (silica; ethyl acetate/hexane ═ 50%) and concentrated to give the desired compound of formula 7-6 (0.077g, 85.4%) as a yellow liquid.
(Compound 829 (S) -3-fluoro-N- (4- (hydroxycarbamoyl) benzyl) -N- (6- (trifluoromethyl) pyridin-2-yl) pyrrolidine-1-carboxamide)
A compound of formula 7-6, methyl ((S) -4- ((3-fluoro-N- (6- (trifluoromethyl) pyridin-2-yl) pyrrolidine-1-carboxamido) methyl) benzoate, 0.100g, 0.235mmol), hydroxylamine (50.0 wt% aqueous solution; 0.144mL, 2.351mmol) and potassium hydroxide (0.132g, 2.351mmol) were dissolved in methanol (1mL) at room temperature and stirred at the same temperature for 1 hour. The solvent was removed from the reaction mixture, and saturated aqueous sodium bicarbonate solution (20mL) was added to the concentrate and stirred. Then, the precipitated solid was filtered, washed with hexane and dried to obtain the desired compound 829(0.051g, 51.1%) as a white solid.
1H NMR(400MHz，CD3OD)d 7.94(d，1H，J＝8.4Hz)，7.89(t，1H，J＝7.7Hz)，7.68(d，1H，J＝8.3Hz)，7.53(d，2H，J＝8.4Hz)，7.37(d，1H，J＝7.4Hz)，7.24(d，1H，J＝8.4Hz)，5.14(s，2H)，3.56～3.38(m，5H)，2.21～1.92(m，2H)；MS(ESI)m/z 427.1(M++H)。
The structural formulae of the compounds are shown in tables 1 to 13.
[ Table 1]
[ Table 2]
[ Table 3]
[ Table 4]
[ Table 5]
[ Table 6]
[ Table 7]
[ Table 8]
[ Table 9]
[ Table 10]
[ Table 11]
[ Table 12]
[ Table 13]
Protocols for measuring and analyzing the Activity of Compounds of the invention
Example 170: determination of inhibition of HDAC Activity (in vitro)
Selective HDAC6 inhibitors are critical for the selectivity of HDAC1 inhibition, which causes side effects and thus determines the selectivity and cellular selectivity of HDAC1/6 enzyme (HDAC 1: histone acetylation/HDAC 6: tubulin acetylation).
1. Testing HDAC Activity (HDAC1 and HDAC6)
The HDAC inhibitory activity of the test material was measured using HDAC1 fluorescent drug detection kit (Enzolife sciences: BML-AK511) and HDAC6 human recombinant (Calbiochem: 382180). In the HDAC1 assay, samples were treated with concentrations of 100, 1000, and 10000nM, and in the HDAC6 assay, samples were treated with concentrations of 0.1, 1, 10, 100, and 1000 nM. After the sample treatment, the sample was reacted at 37 ℃ for 60 minutes, treated with a developer, and then reacted at 37 ℃ for 30 minutes. Then, the fluorescence intensity (Ex 390, Em 460) was measured using flexstation 3(Molecular device).
The results are shown in Table 14.
As shown in table 14, the novel urea derivatives of the present invention exhibited excellent HDAC1/6 enzyme selectivity.
[ TABLE 14] HDAC inhibitory Activity (HDAC1, 6)
Compound (I) HDAC6(uM) HDAC1(uM) Compound (I) HDAC6(uM) HDAC1(uM)
252 0.006 0.151 376 0.004 ND
254 0.002 0.173 377 0.005 ND
256 0.001 0.148 379 0.004 ND
260 0.007 0.551 385 0.005 0.976
261 0.002 0.072 386 0.006 4.04
262 0.001 0.164 389 0.037 7.83
263 0.002 0.075 390 0.020 6.22
279 0.001 0.292 391 0.003 2.12
280 0.005 1.26 392 0.005 2.87
281 0.027 3.91 393 0.005 1.11
309 0.003 ND 394 0.011 2.59
311 0.013 22.9 395 0.005 0.489
312 0.007 22.7 396 0.005 0.257
313 0.007 35.9 397 0.014 1.0
352 0.007 4.62 398 0.032 4.45
353 0.010 4.53 399 0.007 0.302
354 0.009 3.91 400 0.008 0.770
355 0.005 2.34 401 0.008 1.79
356 0.004 3.82 402 0.003 0.170
357 0.003 3.28 403 0.001 0.453
358 0.009 4.17 404 0.016 3.50
370 0.001 0.710 405 0.003 1.45
371 0.020 1.33 413 0.0004 0.860
372 0.001 0.212 414 0.001 1.17
Compound (I) HDAC6(uM) HDAC1(uM) Compound (I) HDAC6(uM) HDAC1(uM)
415 0.005 1.22 465 0.009 2.28
416 0.002 0.930 466 0.003 0.408
418 0.002 0.190 467 0.001 0.240
419 0.005 0.460 468 0.010 12.3
420 0.004 0.310 469 0.012 5.34
438 0.004 1.74 470 0.001 0.635
439 0.002 0.632 471 0.014 1.15
440 0.002 0.988 478 0.009 0.190
441 0.002 0.447 479 0.007 0.280
450 0.005 6.40 480 0.018 1.07
451 0.005 4.81 481 0.015 2.08
453 0.003 0.604 482 0.001 0.680
454 0.005 1.32 483 0.007 0.840
455 0.006 1.98 484 0.002 0.680
456 0.003 1.27 485 0.008 0.860
457 0.005 3.36 486 0.006 1.45
458 0.001 0.474 487 0.007 0.178
459 0.014 7.35 488 0.0005 1.17
460 0.003 2.37 489 0.0005 1.22
461 0.001 2.72 490 0.019 0.703
462 0.014 1.93 491 0.009 3.22
463 0.003 0.284 492 0.002 1.26
464 0.010 1.17 493 0.011 0.883
Compound (I) HDAC6(uM) HDAC1(uM) Compound (I) HDAC6(uM) HDAC1(uM)
494 0.001 0.692 580 0.021 0.667
495 0.011 3.33 651 0.0046 0.765
496 0.001 1.12 683 0.0014 0.545
497 0.002 0.936 684 0.019 4.981
498 0.027 4.71 716 0.0064 0.727
499 0.005 3.63 717 0.024 2.372
500 0.003 1.00 718 0.405 8.541
511 0.012 0.636 765 0.0018 0.433
512 0.014 1.45 766 0.003 0.838
513 0.015 0.654 771 0.0645 ＞10
514 0.015 1.24 772 0.0025 0.349
517 0.028 0.427 773 0.0028 0.312
518 0.014 0.495 774 0.119 12.294
520 0.036 1.172 776 0.0315 1.983
521 0.010 0.261 778 0.011 6.435
522 0.011 0.177 791 0.0011 0.585
529 0.002 0.574 797 0.002 1.601
530 0.0014 0.161 800 0.078 0.633
531 0.0051 1.820 801 0.0597 14.925
532 0.0019 0.577 802 0.0094 1.596
533 0.0045 1.888 803 0.213 5.323
543 0.009 0.447 826 0.0118 1.853
544 0.021 0.503 827 0.0247 1.743
545 0.017 0.545 828 0.0367 4.142
577 0.0012 1.004 829 0.0082 1.603
578 0.0093 1.254
Inhibitory activity of compounds 255 and 374 on all HDACs was confirmed (by Reaction BiologyCorp.). The results are shown in Table 15:
[ Table 15] HDAC inhibitory Activity of Compounds 255 and 374
As shown in table 15, compound 255 was found to be effective against HDAC 6: 0.001uM and HDAC 1: 0.16uM has 160-fold selectivity, and compound 374 is 160-fold selective for HDAC 6: 0.005uM and HDAC 1: 2.12uM had 424 times selectivity.
2. Determination of acetylation of tubulin, Histone H3 and Histone H4 in cells
To determine selectivity for HDAC6 in cells, the acetylation levels of tubulin, histone H3 and histone H4, dependent on compound concentration, were measured by western blot.
Mixing RPMI8226(1.0x 10)6individual cells/well) cells were seeded in a six-well plate and treated with drugs at each concentration (compounds 255 and 374) after 24 hours, proteins were extracted with RIPA buffer and quantified by the Bradford method 25 μ g of protein was dissolved in sample buffer, electrophoresed on a 4-12% gradient gel, then transferred to nitrocellulose membrane for 50 minutes, then the transferred protein was blocked with 5% skim milk for 1 hour, anti-acetyl H3 (1: 2,000), anti-acetyl H4 (1: 5,000), anti-acetyl tubulin (1: 5,000) and anti- β -actin antibody (1: 10,000) were added to 5% skim milk, and the membrane was immersed in skim milk, reacted at 4 ℃ for 16 hours, then washed three times with 1X TBS-T, 10 minutes each time, then IgG-milk HRP antibody (1: 5,000) was added to 5% skim, then the membrane was acetylated at room temperature for 40 minutes, then washed three times with 1X TBS-T, washed for 10 minutes each time, and the level of LAS was determined using 3000 l.
The results are shown in FIGS. 1 and 2.
As shown in fig. 1 and 2, both compound 255 (fig. 1) and compound 374 (fig. 2) were found to have excellent activity at low concentrations, since tubulin acetylation (HDAC6) is expressed at low concentrations of about 10 nM. In contrast, compound 255 and compound 374 were found to have little or no activity at low concentrations, as histone acetylation (HDAC1) was expressed at concentrations of 1uM of compound 255 and 10uM of compound 374. Therefore, it was found from the difference in the expression concentrations of tubulin and histone that the novel urea compound of the present invention has excellent cell selectivity.
Example 170: efficacy of Compounds 254, 255 and 374 in collagen-induced arthritis model
The efficacy of the novel urea derivatives of the invention for the treatment of arthritis in a collagen-induced arthritis model was determined.
The emulsion was prepared by mixing bovine collagen type II (Chondrex) and complete Freund's adjuvant (Chondrex) in a ratio of 1: 1 until the viscosity was measured using a 1ml pipette. Each 100ul of the emulsion was injected intradermally into the tail of DBA/1J mice for the first immunization. After 21 days, an emulsion was prepared by mixing bovine collagen type II and incomplete Freund's adjuvant (Chondrex) in a ratio of 1: 1 in the same manner as described above. Each 100ul of the emulsion was injected intradermally into the upper part of the tail of a DBA/1J mouse for a second booster immunization. After the second booster immunization, mice were grouped according to body weight, test drugs were administered to each group, and the effect of the test drugs was measured by measuring clinical scores and body weights. The clinical score was assigned from 0 to 4, and the total clinical score was determined by evaluating the feet of each mouse and summing the scores (normal: 0, most severe edema: 16). That is, efficacy in the arthritis model was evaluated using the severity of joint edema, and a higher score was assigned to more severe edema.
The results of the above experiments are shown in fig. 3 and 4.
As shown in fig. 3, the clinical score of the vehicle group was 11.6, while the clinical score of the group treated with compound 254 in an amount of 30mg/kg twice a day was 6.4, from which it can be seen that the arthritis symptoms were improved by more than 40%. Furthermore, the clinical scores of the groups treated with compound 255 in an amount of 10 and 30mg/kg twice daily were 5.2 and 6.5, from which it can be seen that the arthritic symptoms were improved by more than 50%. Furthermore, as shown in fig. 4, the clinical score of the vehicle (v) group was 6.7, while the clinical scores of the groups treated with the different concentrations of compound 374 were 2.8 and 2.5, from which it can be seen that the arthritis symptoms were improved by more than 60%. That is, severe edema was observed in the vehicle group which was not treated with the drug, whereas edema was significantly reduced in the group treated with the novel urea compound of the present invention, and thus it can be seen that arthritis symptoms were significantly reduced.
Example 171: efficacy of compounds 255, 374 and 461 in an adjuvant-induced arthritis model
The efficacy of the novel urea derivatives of the invention for the treatment of arthritis in an adjuvant-induced arthritis model was determined.
Complete Freund's adjuvant (Chondrex) containing 10mg/mL of Heat killed tubercular Mycobacteria toxin (Heat killed mycobacterium toxin) was mixed well, and 100uL was then taken and injected intradermally into the upper part of the tail of Lewis rat to induce arthritis. After 10 days, the length of the ankle circumference of the rats was measured, and then the rats were grouped and used for the experiment. Each compound was injected intraperitoneally twice daily. Clinical scores, length and weight of ankle circumference were measured twice weekly after compound administration. Clinical scores were assigned as 0 to 4 based on the arguments of Woods et al (j. immunology, 1214-1222, 2001), and the total clinical score (normal: 0, most severe edema: 16) was determined by evaluating the feet of each rat and summing the scores. After measuring the upper and lower faces (a) of the foot and the lateral face (b) of the foot, the length of the ankle girth was calculated based on the formula of 2X3.14(√ a2+ b 2/2). The final value is determined by averaging the calculated values for each foot.
The results are shown in FIG. 5. In fig. 5, (a) shows the experimental result of compound 255, (B) shows the experimental result of compound 374, and (C) shows the experimental result of compound 461. In fig. 5 (a), the clinical scores of the vehicle group were 4.9, and the clinical scores of the experimental groups treated with compound 255 in amounts of 3, 10 and 30mg/kg twice a day were 3.0, 2.6 and 0.7, from which it can be seen that arthritis symptoms were improved by 40 to 90%. In fig. 5 (B), the clinical scores of the vehicle group were 8.1, and the clinical scores of the experimental groups treated with compound 374 at amounts of 3, 10 and 30mg/kg twice daily were 2.5, 0.8 and 1.3, from which it can be seen that the arthritis symptoms were improved by 70 to 90%. In each experiment, comparison of compounds 255 and 374 with the control compound Tofacitinib (Tofacitinib) showed that they were equally potent. In fig. 5 (C), the clinical score of the vehicle group was 4.2, while the clinical scores of the experimental groups treated with compound 461 in an amount of 25 and 50mg/kg twice a day were 1.9 and 1.3, from which it can be seen that the joint inflammation pattern was improved by more than 50 to 70%, which was higher than MTX used as the control group.
Example 172: efficacy of compounds 254 and 255 in colitis models
The efficacy of the novel urea derivatives of the invention in the treatment of colitis in a colitis model was determined.
Sodium dextran sulfate (DSS: MP biomedicals) 3.0% was dissolved in deionized water and supplied to 9-week-old C57BL/6J mice for 5 to 7 days, and then the mice were normally supplied with water to induce colitis in animal models. The experimental animals were classified into a solvent control group and each group treated with compound 254 and compound 255 as test materials based on the body weight of the experimental animals. The experimental material was injected intraperitoneally twice daily 8 days after DSS administration. Evaluation of efficacy in the colitis model was performed by determining the extent to which the weight lost due to disease was restored to normal levels, and the results are shown in fig. 6.
As shown in fig. 6, the weight loss of the disease-induced model was 17.5 to 18g, then returned to 18.5 to 19.0g by administration of compound 254 and 19 to 19.5g by administration of compound 255, indicating excellent efficacy. As a result, the efficacy of the novel urea derivative for the treatment of colitis was confirmed.
Example 173: anti-cancer activity in multiple myeloma
The efficacy of the novel urea derivatives of the invention for the treatment of multiple myeloma is determined by western blot, MTT assay and combinatorial index.
1. Determination of acetylation of tubulin, Histone H3 and Histone H4
In myeloma-derived RPMI8226 cells (1.0X 10)6Individual cells/well) acetylation of tubulin, histone H3 and histone H4 was determined in the same manner as the cell determination in example 1.
The results are shown in FIGS. 1 and 2.
As shown in fig. 1 and 2, compounds 255 and 374 were found to be potent in the treatment of multiple myeloma with excellent cell selectivity.
2. Determination of cell viability
RPMI8226 cells at 180. mu.l/well at 3X105One cell/ml was seeded in a 96-well U-bottom plate and 20. mu.l of test material was added to the plate. Cells were humidified at 37 ℃ with 5% CO2Incubate for 48 hours and add 20. mu.l each of 5mg/ml 2- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazol-e dissolved in PBSBromide (MTT) solution and incubated for another 4 hours. In the formation of a nail(formazan), the medium was completely removed, 200. mu.l of DMSO was added, and then the absorbance was measured at 570nm using a microplate reader.
3. Determination of Combination Index (CI) for combination therapy
For quantitative analysis of drug interactions between the novel urea compounds of the invention and bortezomib (Velcade) (BOZ), a well-known therapeutic agent for multiple myeloma, the Combination Index (CI) of the cellular fraction for drug-induced cytotoxicity was calculated based on dose-response curves for both single and combination therapies. For the analysis of the interaction between the two drugs, measurements were performed according to the method of Chou and Talalay (CompuSyn by Ting-Chao Chou and Nick Martin, 2005) using Compuyn software (Combosyn Inc.). CI is the dose ratio of the two anti-cancer agents treated with the two anti-cancer doses at the points representing equal toxic effects. CI < 1, CI ═ 1, and CI > 1 are interpreted as synergistic, additive, and antagonistic, respectively.
The results are shown in Table 16 and FIG. 7.
[ Table 16] analysis of combination index for combination therapy
As shown in table 16 and fig. 7, the efficacy of the combination of the two drugs can be determined by the Combination Index (CI), and when this index is not less than 1, the combination effect is excellent. In the case of compound 255, CI was not less than 1 in most doses, and thus it can be seen that the combination effect was excellent.

Claims (8)

1. A compound represented by the following formula I:
[ formula I ]
Wherein
A is
Xa and Xb are each independently of the other C,
L1and L2Each independently is hydrogen, -F, -Cl, -Br or-I,
q is C (═ O),
y is selected from the group consisting of:
m is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4A linear or branched alkyl group, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; a saturated or unsaturated 5-to 7-membered heterocyclic compound comprising from 1 to 3 heteroatoms selected from N, O and S as ring members, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group;wherein R iscAnd RdEach independently is hydrogen, C1-3A linear or branched alkyl group;
n is an integer of 0, 1 or 2,
Rbis hydrogen; a hydroxyl group; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CF3(ii) a -F; -Cl; -Br; -I; orWherein R iseAnd RfEach independently is hydrogen or-C1-3A linear or branched alkyl group,
and is
Z is selected from the group consisting of:
wherein P isaAnd PbEach independently isHydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F; or-C1-3The alcohol is added into the mixture of the alcohol,
whereinSelected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan,Tetrahydropyridine, piperidine or the following groups:
x, y and z are each independently an integer of 0 or 1, and
Rg1、Rg2and Rg3Each independently selected from hydrogen; a hydroxyl group; -C1-3An alkyl group; -CF3；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -N (CH)3)2(ii) a -F; -Cl; -Br; -I; a phenyl group; -S ((═ O)2)CH3(ii) a Or the following groups:
2. the compound of formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound represented by the above formula I is a compound represented by the following formula II:
[ formula II ]
Wherein
a is an integer of 0, 1 or 2,
k is independently hydrogen, -F, -Cl, -Br or-I,
y is selected from the group consisting of:
m is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4Straight or branched chainAn alkanyl group which is unsubstituted or substituted by one or more F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4A linear or branched alkyl group, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; a saturated or unsaturated 5-to 7-membered heterocyclic compound comprising from 1 to 3 heteroatoms selected from N, O and S as ring members, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group;wherein R iscAnd RdEach independently is hydrogen, C1-3A linear or branched alkyl group;
n is an integer of 0, 1 or 2,
Rbis hydrogen; a hydroxyl group; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CF3；-F；-Cl；-Br; -I; orWherein R iseAnd RfEach independently is hydrogen or-C1-3A linear or branched alkyl group, and
z is selected from the group consisting of:
wherein P isaAnd PbEach independently isHydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F or-C1-3The alcohol is added into the mixture of the alcohol,
whereinSelected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan, tetrahydropyridine, piperidine or the following groups:
x, y and z are each independently an integer of 0 or 1, and
Rg1、Rg2and Rg3Each independently selected from hydrogen; a hydroxyl group; -C1-3An alkyl group; -CF3；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -C (═ O) CH3；-C1-4A straight or branched chain alcohol; -N (CH)3)2(ii) a -F; -Cl; -Br; -I; a phenyl group; -S ((═ O)2)CH3(ii) a Or byThe following groups:
3. the compound of formula II according to claim 2, an optical isomer thereof or a pharmaceutically acceptable salt thereof,
wherein
a is an integer of 0, 1 or 2,
k is independently hydrogen, -F, -Cl, -Br or-I,
y is
M is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -C1-4A straight or branched chain alcohol; a benzhydryl group; -C substituted with a saturated or unsaturated 5-to 7-membered heterocyclic compound containing from 1 to 3 heteroatoms selected from N, O and S as ring members1-4A linear or branched alkyl group, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; a saturated or unsaturated 5-to 7-membered heterocyclic compound comprising from 1 to 3 heteroatoms selected from N, O and S as ring members, wherein the heterocyclic compound may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I; phenyl, unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; benzyl which is unsubstituted or substituted by one or more of F, Cl, Br, I, C1-4Alkoxy radical, C1-2Alkyl or hydroxy substitution; -S ((═ O)2)CH3；-F；-Cl；-Br；-I；-C1-6A linear or branched alkoxy group; -C2-6An alkyl alkoxy group; -C (═ O) RxWherein R isxIs straight or branched C1-3Alkyl or C3-10A cycloalkyl group; or
n is an integer of 0, 1 or 2,
Rbis hydrogen; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; and is
Z is selected from the group consisting of:
wherein P isaAnd PbEach independently is hydrogen; a hydroxyl group; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; -F; -Cl; -Br; -I; -CF3；-OCF3；-CN；-C1-6A linear or branched alkoxy group; -C2-6A linear or branched alkylalkoxy group; -CH2F；-C1-3An alcohol;
4. the compound of formula II, an optical isomer thereof, or a pharmaceutically acceptable salt thereof according to claim 2, wherein the compound represented by the above formula II is a compound represented by the following formula III:
[ formula III ]
Wherein
Y is
M is C, O or N, and M is,
l and m are each independently an integer of 0 or 1,
Ra1and Ra2Each independently is hydrogen; -C1-4A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; by phenyl, pyridine or pyrimidine substituted-C1-4A linear or branched alkyl group, wherein the phenyl, pyridine or pyrimidine may be unsubstituted or at least one hydrogen may optionally be replaced by OH, OCH3、CH3、CH2CH3F, Cl, Br or I;
n is an integer of 0, 1 or 2,
Rbis hydrogen; -C1-6A straight or branched chain alkyl group which is unsubstituted or substituted by one or more of F, Cl, Br or I; and is
Z is selected from the group consisting of:
wherein P isaAnd PbEach independently is hydrogen; -F; -Cl; -Br; -CF3；-OCF3；-CH2F；
5. A compound of formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula I is selected from the group consisting of:
6. the compound of formula I, an optical isomer thereof, or a pharmaceutically acceptable salt thereof according to claim 5, wherein the compound of formula I is selected from the group consisting of:
7. a pharmaceutical composition for preventing or treating histone deacetylase-mediated diseases, comprising the compound according to any one of claims 1 to 6, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
8. Use of a compound according to any one of claims 1 to 6, an optical isomer thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of histone deacetylase mediated diseases.
HK16104539.2A 2013-04-29 2014-04-29 Novel compounds for selective histone deacetylase inhibitors, and pharmaceutical composition comprising the same HK1216532B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
KR10-2013-0047212 2013-04-29
KR20130047212 2013-04-29
PCT/KR2014/003776 WO2014178606A1 (en) 2013-04-29 2014-04-29 Novel compounds for selective histone deacetylase inhibitors, and pharmaceutical composition comprising the same

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HK1216532A1 HK1216532A1 (en) 2016-11-18
HK1216532B true HK1216532B (en) 2018-09-28

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