Patent Application
20240327408
The present invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to compounds containing a tetrahydronaphthyridinone or tetrahydropyridopyrimidinone skeleton, a preparation method therefor and pharmaceutical use thereof.
Caseinolytic protease P (ClpP) is an oligomeric serine protease widely present in eukaryotic and prokaryotic cells. ClpP is a major protease in bacteria, which together with another protease LON degrade about 80% of proteins in bacteria, and its function has a key impact on bacterial infectivity. Therefore, early research on ClpP mainly focused on antibacterial drugs. ClpP is an important target for the development of antibacterial drugs against drug-resistant bacterial infections. In human cells, ClpP mainly exists in the mitochondrial matrix, can participate in the degradation of damaged or misfolded proteins in the mitochondrial matrix, and plays a key role in maintaining mitochondrial protein homeostasis. Research in recent years has found that ClpP is a unique target for anti-tumor drugs. Activation of ClpP can promote the selective degradation of ClpP substrates including a variety of respiratory chain proteins, thereby affecting the intracellular oxidative phosphorylation process and leading to the death of malignant tumor cells. ClpP has been reported to be overexpressed in a variety of cancers, including acute myeloid leukemia, breast cancer, lung cancer, liver cancer, ovarian cancer, bladder cancer, prostate cancer, uterine cancer, gastric cancer, testicular cancer, thyroid cancer, etc. (Nat. Rev. Mol. Cell Biol. 2018, 19, 109-120; Biochem. Biophys. Res. Commun. 2017, 491, 85-90; PeerJ 2020, 8, e8754), and ClpP agonists have been reported to have an inhibitory effect on the growth of various tumor cells and induce tumor cell apoptosis. For example, ADEP41 can effectively induce apoptosis in a variety of cancer cell lines, such as HeLa cervical cancer cells, U2OS osteosarcoma cells, and SH-SY5Y undifferentiated neuroblastoma cells (Cell Chem. Biol. 2018, 25, 1017-1030). In addition, the ClpP agonist ONC201 can cause tumor regression in multiple xenograft solid tumor models such as colon cancer, breast cancer, and brain tumors (Oncotarget 2016, 7, 74380-74392).
Objective: The present invention aims to provide new ClpP agonists, i.e., compounds containing a tetrahydronaphthyridinone or tetrahydropyridopyrimidinone skeleton, which have significant agonistic effect on caseinolytic protease P (ClpP).
Technical scheme: provided are compounds of formula I or pharmaceutically acceptable salts thereof according to the present invention, wherein
Preferably, R2 is selected from cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, benzyl, 2-fluorobenzyl, 2-chlorobenzyl, 2-bromobenzyl, 2-methylbenzyl, 2-ethylbenzyl, 2-methoxybenzyl, 3-fluorobenzyl, 3-chlorobenzyl, 3-bromobenzyl, 3-methylbenzyl, 3-ethylbenzyl, 3-methoxybenzyl, 4-fluorobenzyl, 4-chlorobenzyl, 4-bromobenzyl, 4-methylbenzyl, 4-ethylbenzyl, 4-methoxybenzyl, 4-nitrobenzyl, 4-methylaminobenzyl, 4-dimethylaminobenzyl, 4-trifluoromethylbenzyl, 2,4-difluorobenzyl, 2-fluoro-4-chlorobenzyl, 2-fluoro-4-methylbenzyl 2-fluoro-4-methoxybenzyl, 2-fluoro-4-trifluoromethylbenzyl, 2-fluoro-4-bromobenzyl, (1-methyl-1H-pyrazol-3-yl)methyl, (1-methyl-1H-pyrazol-5-yl)methyl, (1,3-dimethyl-1H-pyrazol-5-yl)methyl, pyridin-2-ylmethyl, pyridin-3-ylmethyl, pyridin-4-ylmethyl, (4-chloropyridin-2-yl)methyl, (1-methyl-1H-imidazol-2-yl)methyl, (1-methyl-1H-imidazol-4-yl)methyl, (1-methyl-1H-imidazol-5-yl)methyl, (3,5-dimethylisoxazol-4-yl)methyl, (2-methylthiazol-5-yl)methyl, benzo[1,3]dioxol-4-ylmethyl, benzo[1,3]dioxol-5-ylmethyl, benzo[1,4]dioxan-5-ylmethyl, benzo[1,4]dioxan-6-ylmethyl, and the like.
As a preferred technical scheme, the compounds of the present invention are shown in the following formula:
Further, the preferred compounds of formula I of the present invention are as follows:
Provided is a preparation method for the compounds represented by formula I described above, wherein when ring A is of a structure represented by general formula II, a synthetic route comprises
When ring A in general formula I is an imidazole ring in general formula III, a synthetic route comprises
When ring A in the general formula I is an imidazole ring, a triazole ring, a pyrazole ring or a pyrrole ring in general formula IV, and X, Y and Z are each a nitrogen atom or a methine group, a synthetic route comprises
When ring A in general formula I is an imidazole ring, a triazole ring or a pyrazole ring in general formula IV, and X, Y and Z are each a nitrogen atom or a methine group, a synthetic route may also comprise
When R2 in general formula IV does not contain a group that can be reduced by hydrogenation, a synthetic route may also comprise
Provided are pharmaceutical compositions according to the present invention, which comprise the compounds or the pharmaceutically acceptable salts thereof described above.
Use of the compounds of formula I or the pharmaceutically acceptable salts thereof described above and the pharmaceutical compositions described above in the preparation of ClpP agonists are also within the protection scope of the present invention.
Use of the compounds of formula I or the pharmaceutically acceptable salts thereof described above and the pharmaceutical compositions described above in the preparation of medicaments for treating cancer are also within the protection scope of the present invention. The drugs achieve the purpose of treating cancer by activating caseinolytic protease P (ClpP).
Beneficial effects: Through experimental verification, the compounds containing a tetrahydronaphthyridinone or tetrahydropyridopyrimidinone skeleton of the present invention have significant agonistic effects on caseinolytic protease P (ClpP) and can be applied to the treatment of various cancers.
The content of the present invention will be specifically described below through examples.
Ethyl 1-benzyl-4-piperidone-3-carboxylate hydrochloride (1) (10 g, 33.6 mmol) was dissolved in 200 mL of ethanol at room temperature, and palladium on carbon (1.0 g, 10% w/w) was added. The reaction system was purged with nitrogen, followed by hydrogen, and then subjected to a hydrogenation reaction at 1 atm for 12 h. After the reaction was completed as monitored by TLC, di-tert-butyl dicarbonate (8.1 g, 37.0 mmol) and triethylamine (10.3 mL, 73.9 mmol) were added, and the mixture was stirred for reaction at room temperature for 8 h. After the reaction was completed as monitored by TLC, the palladium on carbon was filtered out through celite, and the filtrate was concentrated by rotary evaporation under reduced pressure to remove the solvent. 200 mL of water was added, and the resulting mixture was extracted with dichloromethane (150 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography to give a colorless oily liquid (8.6 g, yield: 94%). 1H NMR (300 MHz, Chloroform-d) δ 4.23 (q, J=6.8 Hz, 2H), 4.08 (s, 2H), 3.28 (t, J=5.8 Hz, 2H), 2.37 (t, J=5.8 Hz, 2H), 1.58 (s, 9H), 1.32 (t, J=6.8 Hz, 3H).
Ethyl 1-N-tert-butoxycarbonyl-4-piperidone-3-carboxylate 2 (7.5 g, 27.7 mmol) was dissolved in 150 mL of dichloromethane at room temperature, and N,N-diisopropylethylamine (11.7 mL, 82.8 mmol) was added under argon atmosphere. The reaction system was cooled to −78° C., and trifluoromethanesulfonic anhydride (6.0 mL, 35.9 mmol) was slowly added dropwise. After stirring for 1 h, the reaction system was warmed to 0° C., and stirred at this temperature for 23 h. After the reaction was completed as monitored by TLC, the reaction was quenched by adding 100 mL of saturated sodium bicarbonate solution. The aqueous phase was extracted with dichloromethane (50 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography to give a colorless oily liquid (6.5 g, yield: 58%). 1H NMR (300 MHz, Chloroform-d) δ 4.38-4.24 (m, 4H), 3.63 (t, J=5.7 Hz, 2H), 2.57-2.46 (m, 2H), 1.49 (s, 9H), 1.34 (t, J=7.1 Hz, 3H).
1-(Tert-butyl)-3-ethyl-4-(((trifluoromethyl)sulfonyl)oxy)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate 3 (6.5 g, 16.1 mmol) was dissolved in 130 mL of 1,4-dioxane at room temperature, and bis(pinacolato)diboron (4.9 g, 19.4 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.18 g, 1.61 mmol), 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl (661 mg, 1.61 mmol), and potassium acetate (3.16 g, 32.2 mmol) were added under argon atmosphere. The mixture was stirred and reacted at reflux overnight. After the reaction was completed as monitored by TLC, the reaction mixture was cooled to room temperature, filtered to remove the solid, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography to give a colorless oily liquid (3.6 g, yield: 54%). 1H NMR (300 MHz, Chloroform-d) δ 4.20-3.98 (m, 4H), 3.36 (t, J=5.4 Hz, 2H), 2.28-2.20 (m, 2H), 1.38 (s, 9H), 1.26 (s, 12H), 1.24 (t, J=6.5 Hz, 3H).
1-(Tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) (3 g, 7.87 mmol) was dissolved in a mixed solvent of 48 mL of 1,4-dioxane and 8 mL of water at room temperature, and 2-amino-3-iodopyridine (2.60 g, 11.8 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (578 mg, 0.79 mmol), 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl (324 mg, 0.79 mmol), and cesium carbonate (3.84 g, 11.8 mmol) were added. The mixture was stirred and reacted at reflux overnight under argon atmosphere. After the reaction was completed as monitored by TLC, the reaction mixture was cooled to room temperature, filtered to remove the solid, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography to give a white solid (1.9 g, yield: 80%). 1H NMR (300 MHz, Chloroform-d) δ 13.16 (s, 1H), 8.75 (d, J=3.6 Hz, 1H), 8.00 (d, J=7.7 Hz, 1H), 7.26 (dd, J=7.9, 4.8 Hz, 1H), 4.51 (s, 2H), 3.78 (t, J=5.6 Hz, 2H), 2.93 (t, J=5.7 Hz, 2H), 1.52 (s, 9H).
Compound 5-B was synthesized by referring to the preparation method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylic acid (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) and 2-iodoaniline as starting materials. White solid, 1H NMR (300 MHz, Chloroform-d) δ 12.54 (s, 1H), 8.42 (d, J=3.6 Hz, 1H), 7.62 (d, J=7.7 Hz, 1H), 7.13 (dd, J=7.9, 4.8 Hz, 1H), 4.46 (s, 2H), 3.82 (t, J=5.5 Hz, 2H), 2.91 (t, J=5.6 Hz, 2H), 1.52 (s, 9H).
Compound 5-C was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridinedicarboxylate (4) and 4-amino-5-iodopyrimidine as starting materials. White solid, 1H NMR (300 MHz, Chloroform-d) δ 11.27 (s, 1H), 9.13 (s, 1H), 9.00 (s, 1H), 4.52 (s, 2H), 3.79 (t, J=5.7 Hz, 2H), 3.00 (t, J=5.5 Hz, 2H), 1.51 (s, 9H).
Compound 5-D was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) and 3-amino-4-iodofuran as starting materials. White solid, MS (ESI) m/z: 291.2 [M+H]+.
Compound 5-E was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) and 3-amino-4-iodothiophene as starting materials. White solid, MS (ESI) m/z: 307.2 [M+H]+.
Compound 5-F was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) and 3-amino-2-iodothiophene as starting materials. White solid, MS (ESI) m/z: 307.2 [M+H]+.
Compound 5-G was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridine dicarboxylate (4) and 2-amino-3-iodothiophene as starting materials. White solid, MS (ESI) m/z: 307.2 [M+H]+.
Compound 5-H was synthesized by referring to the synthetic method for the compound tert-butyl 5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3-(2H)-carboxylate (5-A) in Example 4, with 1-(tert-butyl)-3-ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-5,6-dihydro-1,3(2H)-pyridinedicarboxylate (4) and 4-amino-5-iodothiazole as starting materials. White solid, MS (ESI) m/z: 308.2 [M+H]+.
Compound 5-A-1 (600 mg, 1.99 mmol) was dissolved in 10 mL of N,N-dimethylformamide at room temperature, and potassium carbonate (275 mg, 3.98 mmol) was added, followed by dropwise addition of benzyl bromide (681 mg, 3.98 mmol). The mixture was stirred overnight. After the reaction was completed as monitored by TLC, 50 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography to give a white solid (545 mg, yield: 70%). 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.7 Hz, 1H), 7.91 (dd, J=7.9, 1.7 Hz, 1H), 7.48-7.43 (m, 2H), 7.27 (d, J=4.3 Hz, 1H), 7.24-7.13 (m, 3H), 5.77 (s, 2H), 3.58 (s, 2H), 2.93 (t, J=5.6 Hz, 2H), 2.78 (t, J=5.7 Hz, 2H), 1.43 (s, 9H); MS (ESI) m/z: 392.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and (bromomethyl)cyclopropane as starting materials. MS (ESI) m/z: 356.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and (bromomethyl)cyclobutane as starting materials. MS (ESI) m/z: 370.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and (bromomethyl)cyclopentane as starting materials. MS (ESI) m/z: 384.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and (bromomethyl)cyclohexane as starting materials. MS (ESI) m/z: 398.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-fluorobenzyl bromide as starting materials. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chlorobenzyl bromide as starting materials. MS (ESI) m/z: 426.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-bromobenzyl bromide as starting materials. MS (ESI) m/z: 471.1 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-methylbenzyl bromide as starting materials. MS (ESI) m/z: 406.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 420.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 422.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-fluorobenzyl bromide as starting materials. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chlorobenzyl bromide as starting materials. MS (ESI) m/z: 426.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-bromobenzyl bromide as starting materials. MS (ESI) m/z: 471.1 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-methylbenzyl bromide as starting materials. MS (ESI) m/z: 406.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 420.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 422.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-fluorobenzyl bromide as starting materials. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chlorobenzyl bromide as starting materials. MS (ESI) m/z: 426.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-bromobenzyl bromide as starting materials. MS (ESI) m/z: 471.1 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-methylbenzyl bromide as starting materials. MS (ESI) m/z: 406.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 420.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-ethylbenzyl bromide as starting materials. MS (ESI) m/z: 422.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 4-trifluoromethylbenzyl bromide as starting materials. MS (ESI) m/z: 460.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 3-bromomethyl-1-methyl-1H-pyrazole as starting materials. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 5-bromomethyl-1-methyl-1H-pyrazole as starting materials. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 5-bromomethyl-1,3-dimethyl-1H-pyrazole as starting materials. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chloromethylpyridine as starting materials. MS (ESI) m/z: 393.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 3-chloromethylpyridine as starting materials. MS (ESI) m/z: 393.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 4-chloromethylpyridine as starting materials. MS (ESI) m/z: 393.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chloromethyl-4-chloropyridine as starting materials. MS (ESI) m/z: 427.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 2-chloromethyl-1-methyl-1H-imidazole as starting materials. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 4-chloromethyl-1-methyl-1H-imidazole as starting materials. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 5-chloromethyl-1-methyl-1H-imidazole as starting materials. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 4-chloromethyl-3,5-dimethylisoxazole as starting materials. MS (ESI) m/z: 411.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 5-chloromethyl-2-methylthiazole as starting materials. MS (ESI) m/z: 413.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 4-nitrobenzyl bromide as starting materials. MS (ESI) m/z: 437.4 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 6-bromomethyl-2,3-dihydro[1,4]benzodioxine as starting materials. MS (ESI) m/z: 450.4 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-A and 5-bromomethyl-benzo[1,3]dioxolane as starting materials. MS (ESI) m/z: 436.3 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-B and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 459.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-C and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 461.2 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-D and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 449.3 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-E and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 465.3 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-F and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 465.3 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-G and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 465.3 [M+H]+.
The compound was prepared by referring to the preparation method for compound 6-A-1 in Example 12, with compound 5-H and 4-(trifluoromethyl)benzyl bromide as starting materials. MS (ESI) m/z: 466.3 [M+H]+.
Tert-butyl 6-benzyl-5-oxo-1,4,5,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-3(2H)-carboxylate (6-A-1) (500 mg, 1.28 mmol) was dissolved in 3 mL of dichloromethane at room temperature, and 6 mL of trifluoroacetic acid was added. The mixture was stirred at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove the solvent, neutralized with a sodium bicarbonate solution to pH 8, extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated to give a yellow viscous liquid (335 mg, yield: 90%); MS (ESI) m/z: 292.2 [M+H]+.
100 mg (0.26 mmol) of 6-benzyl-2,3,4,6-tetrahydropyrido[3,4-c][1,8]naphthyridine-5-(1H)-one (7-1) was dissolved in 1 mL of N,N-dimethylformamide, and 50 μL (0.51 mmol) of (bromomethyl)cyclopropane and 140 μL (0.77 mmol) of N,N-diisopropylethylamine were added. The reaction was stirred at room temperature overnight. After the reaction was completed as monitored by TLC, 5 mL of water was added, and the mixture was extracted with ethyl acetate (5 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by rotary evaporation under reduced pressure to remove the solvent, and purified by silica gel column chromatography. The resulting crude product was purified by semi-preparative HPLC, and freeze-dried to give a white solid (68.8 mg, yield: 78%). 1H NMR (300 MHz, Chloroform-d) δ 8.49 (dd, J=4.6, 1.5 Hz, 1H), 7.81 (dd, J=7.9, 1.5 Hz, 1H), 7.49-7.43 (m, 2H), 7.24-7.05 (m, 4H), 5.75 (s, 2H), 3.65 (s, 2H), 2.87 (t, J=4.6 Hz, 2H), 2.81 (t, J=4.7 Hz, 2H), 2.47 (d, J=6.6 Hz, 2H), 1.03-0.91 (m, 1H), 0.61-0.53 (m, 2H), 0.23-0.16 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 161.41, 148.72, 148.07, 139.34, 137.88, 131.66, 128.48, 128.19, 127.70, 127.02, 118.03, 115.79, 63.36, 51.40, 49.26, 44.01, 25.55, 8.66, 4.10. MS(ESI) m/z: 346.2 [M+H]+, 368.1 [M+Na]+.
The compound was prepared from compound 7-1 and (bromomethyl)cyclobutane by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.7 Hz, 1H), 7.90 (dd, J=7.9, 1.7 Hz, 1H), 7.47-7.40 (m, 2H), 7.26-7.13 (m, 4H), 5.77 (s, 2H), 3.54 (s, 2H), 2.92 (t, J=5.5 Hz, 2H), 2.76 (t, J=5.7 Hz, 2H), 2.71-2.60 (m, 3H), 2.18-2.06 (m, 2H), 1.96-1.71 (m, 4H). 13C NMR (75 MHz, CDCl3) δ 161.50, 148.78, 148.19, 139.26, 137.87, 131.66, 128.47, 128.23, 127.81, 127.03, 118.03, 115.92, 64.84, 51.53, 49.49, 44.05, 33.77, 27.74, 25.52, 18.86. MS (ESI) m/z: 360.2 [M+H]+, 382.3 [M+Na]+.
The compound was prepared from compound 7-1 and (bromomethyl)cyclopentane by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.7 Hz, 1H), 7.91 (dd, J=7.9, 1.7 Hz, 1H), 7.48-7.44 (m, 2H), 7.28-7.25 (m, 1H), 7.23-7.14 (m, 3H), 5.77 (s, 2H), 3.58 (s, 2H), 2.93 (t, J=5.6 Hz, 2H), 2.78 (t, J=5.7 Hz, 2H), 2.50 (d, J=7.4 Hz, 2H), 2.29-2.17 (m, 1H), 1.85-1.75 (m, 2H), 1.64-1.50 (m, 4H), 1.28-1.19 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 161.57, 148.73, 148.20, 139.38, 137.91, 131.67, 128.57, 128.22, 127.96, 127.05, 118.02, 115.97, 64.32, 51.79, 49.65, 44.05, 37.31, 31.38, 25.61, 25.22. MS (ESI) m/z: 374.2 [M+H]+, 396.1 [M+Na]+.
The compound was prepared from compound 7-1 and (bromomethyl)cyclohexane by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.7 Hz, 1H), 7.92 (dd, J=7.9, 1.7 Hz, 1H), 7.49-7.41 (m, 2H), 7.28-7.15 (m, 4H), 5.77 (s, 2H), 3.53 (s, 2H), 2.93 (t, J=5.6 Hz, 2H), 2.74 (t, J=5.7 Hz, 2H), 2.37 (d, J=7.1 Hz, 2H), 1.84-1.59 (m, 7H), 1.29-1.23 (m, 2H), 1.00-0.83 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 161.58, 148.73, 148.21, 139.41, 137.90, 131.67, 128.60, 128.22, 128.01, 127.06, 118.02, 115.99, 65.58, 58.44, 51.99, 49.82, 44.06, 35.16, 31.90, 26.80, 26.13, 25.65, 18.47. MS (ESI) m/z: 388.2 [M+H]+.
The compound was prepared from compound 7-1 and benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.7 Hz, 1H), 7.87 (dd, J=7.9, 1.7 Hz, 1H), 7.48-7.27 (m, 7H), 7.25-7.12 (m, 4H), 5.75 (s, 2H), 3.75 (s, 2H), 3.62 (s, 2H), 2.89 (t, J=5.4 Hz, 2H), 2.77 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.45, 148.82, 148.21, 139.42, 137.89, 137.78, 131.73, 129.23, 128.61, 128.44, 128.24, 127.84, 127.34, 127.09, 118.06, 115.92, 62.65, 51.74, 48.65, 44.08, 25.70. MS (ESI) m/z: 382.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.57 (dd, J=4.6, 1.5 Hz, 1H), 7.89 (dd, J=7.9, 1.5 Hz, 1H), 7.57-7.47 (m, 3H), 7.39 (dd, J=7.5, 1.6 Hz, 1H), 7.31-7.14 (m, 6H), 5.79 (s, 2H), 3.89 (s, 2H), 3.72 (s, 2H), 2.96-2.84 (m, 4H). 13C NMR (75 MHz, CDCl3) δ 161.41, 148.83, 148.20, 139.38, 137.91, 135.62, 134.45, 131.72, 130.85, 129.60, 128.65, 128.45, 128.25, 127.76, 127.11, 126.81, 118.07, 115.88, 59.04, 51.64, 48.96, 44.09, 25.68; MS (ESI) m/z: 416.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The compound was prepared from compound 7-1 and 2-methylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-ethylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-methoxybenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 412.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.58 (dd, J=4.7, 1.7 Hz, 1H), 7.90 (dd, J=7.9, 1.7 Hz, 1H), 7.53-7.46 (m, 2H), 7.42 (s, 1H), 7.30-7.15 (m, 7H), 5.79 (s, 2H), 3.74 (s, 2H), 3.63 (s, 2H), 2.93 (t, J=5.5 Hz, 2H), 2.79 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.39, 148.89, 148.18, 140.12, 139.39, 137.86, 134.33, 131.75, 129.70, 129.03, 128.64, 128.25, 127.58, 127.52, 127.22, 127.12, 118.11, 115.84, 62.00, 51.64, 48.77, 44.09, 25.65; MS (ESI) m/z: 416.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The compound was prepared from compound 7-1 and 3-methylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-ethylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-methoxybenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.7 Hz, 1H), 7.90 (dd, J=7.9, 1.7 Hz, 1H), 7.47-7.43 (m, 2H), 7.25-7.14 (m, 5H), 6.98-6.94 (m, 2H), 6.83-6.79 (m, 1H), 5.76 (s, 2H), 3.80 (s, 3H), 3.73 (s, 2H), 3.63 (s, 2H), 2.91 (t, J=5.5 Hz, 2H), 2.77 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.46, 159.74, 148.82, 148.22, 139.52, 139.41, 137.88, 131.72, 129.38, 128.60, 128.24, 127.87, 127.08, 121.50, 118.05, 115.93, 114.50, 112.84, 62.54, 55.25, 51.80, 48.58, 44.07, 25.70; MS (ESI) m/z: 412.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.50 (dd, J=4.6, 1.7 Hz, 1H), 7.78 (dd, J=7.9, 1.7 Hz, 1H), 7.49-7.41 (m, 2H), 7.29-7.05 (m, 8H), 5.72 (s, 2H), 3.65 (s, 2H), 3.55 (s, 2H), 2.80 (t, J=5.1 Hz, 2H), 2.70 (t, J=5.2 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.36, 148.85, 148.13, 139.40, 137.91, 136.51, 132.96, 131.75, 130.45, 128.72, 128.56, 128.25, 127.57, 127.14, 118.11, 115.80, 61.84, 51.61, 48.76, 44.08, 25.64; MS (ESI) m/z: 416.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The compound was prepared from compound 7-1 and 4-methylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-ethylbenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-methoxybenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 412.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-chloromethyl-1-methyl-1H-pyrazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 385.2 [M+H]+.
The compound was prepared from compound 7-1 and 5-chloromethyl-1-methyl-1H-pyrazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The compound was prepared from compound 7-1 and 5-chloromethyl-1,3-methyl-1H-pyrazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-chloromethylpyridine by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.75-8.72 (m, 1H), 8.70 (dd, J=4.7, 1.7 Hz, 1H), 8.28 (dd, J=8.0, 1.7 Hz, 1H), 7.63-7.58 (m, 1H), 7.55-7.49 (m, 1H), 7.42 (dd, J=8.0, 4.7 Hz, 1H), 7.38-7.32 (m, 2H), 7.26-7.16 (m, 3H), 5.78 (s, 2H), 4.75 (s, 2H), 4.42 (s, 2H), 3.77 (t, J=6.2 Hz, 2H), 3.43 (t, J=6.1 Hz, 2H). MS (ESI) m/z: 383.2 [M+H]+.
The compound was prepared from compound 7-1 and 3-chloromethylpyridine by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.94-8.90 (m, 1H), 8.83-8.78 (m, 1H), 8.68 (dd, J=4.7, 1.7 Hz, 1H), 8.38 (dt, J=8.0, 1.8 Hz, 1H), 8.25 (dd, J=8.0, 1.7 Hz, 1H), 7.82 (dd, J=7.9, 5.3 Hz, 1H), 7.40 (dd, J=8.0, 4.7 Hz, 1H), 7.35-7.27 (m, 2H), 7.25-7.14 (m, 3H), 5.76 (s, 2H), 4.67 (s, 2H), 4.24 (s, 2H), 3.70 (t, J=6.1 Hz, 2H), 3.38 (t, J=6.1 Hz, 2H). MS (ESI) m/z: 383.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-chloromethylpyridine by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.93-8.87 (m, 2H), 8.66 (dd, J=4.7, 1.6 Hz, 1H), 8.24 (dd, J=8.0, 1.7 Hz, 1H), 8.17-8.08 (m, 2H), 7.39 (dd, J=8.0, 4.7 Hz, 1H), 7.33-7.28 (m, 2H), 7.24-7.15 (m, 3H), 5.76 (s, 2H), 4.67 (s, 2H), 4.17 (s, 2H), 3.59 (t, J=6.0 Hz, 2H), 3.38-3.34 (m, 2H). MS (ESI) m/z: 383.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-chloromethyl-4-chloropyridine by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.73 (dd, J=4.7, 1.7 Hz, 1H), 8.67 (d, J=5.4 Hz, 1H), 8.30 (dd, J=8.0, 1.7 Hz, 1H), 7.69 (d, J=1.6 Hz, 1H), 7.60 (dd, J=5.4, 2.0 Hz, 1H), 7.44 (dd, J=8.0, 4.7 Hz, 1H), 7.40-7.35 (m, 2H), 7.29-7.18 (m, 3H), 5.80 (s, 2H), 4.77 (s, 2H), 4.44 (s, 2H), 3.79 (t, J=6.1 Hz, 2H), 3.44 (t, J=6.2 Hz, 2H) MS (ESI) m/z: 417.2 [M+H]+.
The compound was prepared from compound 7-1 and 2-chloromethyl-1-methyl-1H-imidazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-chloromethyl-1-methyl-1H-imidazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The compound was prepared from compound 7-1 and 5-chloromethyl-1-methyl-1H-imidazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The compound was prepared from compound 7-1 and 4-chloromethyl-3,5-dimethylisoxazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 401.2 [M+H]+.
The compound was prepared from compound 7-1 and 5-chloromethyl-2-methylthiazole by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 403.2 [M+H]+.
The Boc protecting group in compound 6-A-2 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.7 Hz, 1H), 7.91 (dd, J=7.9, 1.7 Hz, 1H), 7.41-7.28 (m, 5H), 7.16 (dd, J=7.9, 4.7 Hz, 1H), 4.43 (d, J=7.1 Hz, 2H), 3.77 (s, 2H), 3.63 (s, 2H), 2.92 (t, J=5.5 Hz, 2H), 2.79 (t, J=5.7 Hz, 2H), 1.44-1.35 (m, 1H), 0.55-0.48 (m, 2H), 0.46-0.38 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 161.61, 148.65, 148.39, 139.07, 137.81, 131.61, 129.21, 128.41, 127.78, 127.30, 117.75, 115.85, 62.61, 51.81, 48.61, 45.20, 25.64, 10.35, 3.96, 3.86; MS (ESI) m/z: 346.2 [M+H]+.
The Boc protecting group in compound 6-A-3 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.7 Hz, 1H), 7.90 (dd, J=7.9, 1.7 Hz, 1H), 7.41-7.27 (m, 5H), 7.16 (dd, J=7.9, 4.7 Hz, 1H), 4.61 (d, J=7.3 Hz, 2H), 3.77 (s, 2H), 3.63 (s, 2H), 2.95-2.84 (m, 3H), 2.79 (t, J=5.7 Hz, 2H), 1.96-1.80 (m, 6H). 13C NMR (75 MHz, CDCl3) δ 161.70, 148.57, 148.49, 138.92, 137.81, 131.54, 129.24, 128.41, 127.75, 127.30, 117.70, 115.77, 62.64, 51.85, 48.60, 45.57, 34.80, 26.34, 25.65, 18.42; MS (ESI) m/z: 360.2 [M+H]+.
The Boc protecting group in compound 6-A-4 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.7 Hz, 1H), 7.89 (dd, J=7.9, 1.8 Hz, 1H), 7.41-7.23 (m, 5H), 7.15 (dd, J=7.9, 4.6 Hz, 1H), 4.52 (d, J=7.5 Hz, 2H), 3.77 (s, 2H), 3.63 (s, 2H), 2.92 (t, J=5.5 Hz, 2H), 2.79 (t, J=5.7 Hz, 2H), 2.53 (p, J=7.4 Hz, 1H), 1.72-1.56 (m, 4H), 1.53-1.34 (m, 4H). 13C NMR (75 MHz, CDCl3) δ 161.75, 148.55, 138.93, 137.75, 131.53, 129.25, 128.41, 127.69, 127.31, 117.68, 115.77, 62.64, 51.81, 48.62, 45.21, 39.04, 30.31, 25.62, 24.93; MS (ESI) m/z: 374.2 [M+H]+.
The Boc protecting group in compound 6-A-5 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.53 (dd, J=4.6, 1.7 Hz, 1H), 7.88 (dd, J=7.9, 1.7 Hz, 1H), 7.41-7.23 (m, 5H), 7.14 (dd, J=7.9, 4.6 Hz, 1H), 4.41 (d, J=7.3 Hz, 2H), 3.76 (s, 2H), 3.62 (s, 2H), 2.90 (t, J=5.3 Hz, 2H), 2.78 (t, J=5.6 Hz, 2H), 2.06-1.85 (m, 1H), 1.69-1.54 (m, 4H), 1.28-0.99 (m, 6H). 13C NMR (75 MHz, CDCl3) δ 161.75, 148.63, 148.56, 138.97, 137.78, 131.52, 129.24, 128.40, 127.63, 127.29, 117.67, 115.69, 62.68, 51.79, 48.65, 46.75, 36.71, 30.87, 26.47, 25.98, 25.64; MS (ESI) m/z: 388.2 [M+H]+.
The Boc protecting group in compound 6-A-6 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The Boc protecting group in compound 6-A-7 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 416.2 [M+H]+.
The Boc protecting group in compound 6-A-8 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The Boc protecting group in compound 6-A-9 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.48 (dd, J=4.6, 1.7 Hz, 1H), 7.94 (dd, J=7.9, 1.7 Hz, 1H), 7.41-7.25 (m, 5H), 7.20-7.13 (m, 2H), 7.07 (t, J=6.9 Hz, 1H), 6.95 (t, J=7.5 Hz, 1H), 6.64 (d, J=7.6 Hz, 1H), 5.72 (s, 2H), 3.77 (s, 2H), 3.65 (s, 2H), 2.97 (t, J=5.6 Hz, 2H), 2.82 (t, J=5.7 Hz, 2H), 2.50 (s, 3H). 13C NMR (75 MHz, CDCl3) δ 161.48, 149.07, 148.34, 139.56, 137.68, 135.57, 135.47, 131.75, 130.10, 129.28, 128.44, 127.80, 127.36, 126.57, 125.87, 125.12, 118.10, 115.85, 62.67, 51.75, 48.63, 41.99, 25.75, 19.45; MS (ESI) m/z: 396.2 [M+H]+.
The Boc protecting group in compound 6-A-10 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The Boc protecting group in compound 6-A-11 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 412.2 [M+H]+.
The Boc protecting group in compound 6-A-12 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The Boc protecting group in compound 6-A-13 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 416.2 [M+H]+.
The Boc protecting group in compound 6-A-14 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The Boc protecting group in compound 6-A-15 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 396.2 [M+H]+.
The Boc protecting group in compound 6-A-16 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The Boc protecting group in compound 6-A-17 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 412.2 [M+H]+.
The Boc protecting group in compound 6-A-18 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.2 [M+H]+.
The Boc protecting group in compound 6-A-19 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 416.2 [M+H]+.
The Boc protecting group in compound 6-A-20 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 460.1 [M+H]+.
The Boc protecting group in compound 6-A-21 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 396.2 [M+H]+.
The Boc protecting group in compound 6-A-22 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 410.2 [M+H]+.
The Boc protecting group in compound 6-A-23 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 412.2 [M+H]+.
The Boc protecting group in compound 6-A-25 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The Boc protecting group in compound 6-A-26 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The Boc protecting group in compound 6-A-27 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 400.1 [M+H]+.
The Boc protecting group in compound 6-A-28 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.71-8.66 (m, 1H), 8.60 (dd, J=4.7, 1.6 Hz, 1H), 8.37-8.26 (m, 2H), 7.85-7.74 (m, 2H), 7.65-7.58 (m, 2H), 7.56-7.48 (m, 3H), 7.42 (dd, J=8.0, 4.7 Hz, 1H), 6.03 (s, 2H), 4.61 (s, 2H), 4.27 (s, 2H), 3.75 (s, 2H), 3.41 (t, J=5.7 Hz, 2H); MS (ESI) m/z: 383.2 [M+H]+.
The Boc protecting group in compound 6-A-29 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 9.01 (s, 1H), 8.75 (d, J=5.6 Hz, 1H), 8.72-8.64 (m, 2H), 8.27 (dd, J 10=8.0, 1.5 Hz, 1H), 8.00 (dd, J=8.0, 5.8 Hz, 1H), 7.66-7.57 (m, 2H), 7.53-7.47 (m, 3H), 7.42 (dd, J=8.0, 4.7 Hz, 1H), 5.92 (s, 2H), 4.62 (s, 2H), 4.29 (s, 2H), 3.75 (s, 2H), 3.41 (t, J=5.2 Hz, 2H); MS (ESI) m/z: 383.2 [M+H]+.
The Boc protecting group in compound 6-A-30 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.74 (d, J=6.7 Hz, 2H), 8.58 (dd, J=4.7, 1.5 Hz, 1H), 8.30 (dd, J=8.0, 1.6 Hz, 1H), 7.96 (d, J=6.7 Hz, 2H), 7.66-7.58 (m, 2H), 7.54-7.48 (m, 3H), 7.42 (dd, J=8.0, 4.7 Hz, 1H), 6.00 (s, 2H), 4.62 (s, 2H), 4.30 (s, 2H), 3.77 (s, 2H), 3.44 (t, J=5.6 Hz, 2H); MS (ESI) m/z: 383.2 [M+H]+.
The Boc protecting group in compound 6-A-31 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Methanol-d4) δ 8.56 (dd, J=4.7, 1.6 Hz, 1H), 8.43-8.32 (m, 1H), 8.26 (dd, J=8.0, 1.6 Hz, 1H), 7.66-7.58 (m, 2H), 7.54-7.47 (m, 3H), 7.45-7.41 (m, 2H), 7.38 (dd, J=8.0, 4.7 Hz, 1H), 5.88 (s, 2H), 4.62 (s, 2H), 4.31 (s, 2H), 3.77 (s, 2H), 3.43 (t, J=5.3 Hz, 2H); MS (ESI) m/z: 417.1 [M+H]+.
The Boc protecting group in compound 6-A-32 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The Boc protecting group in compound 6-A-33 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The Boc protecting group in compound 6-A-34 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 386.2 [M+H]+.
The Boc protecting group in compound 6-A-35 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 401.2 [M+H]+.
The Boc protecting group in compound 6-A-36 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 403.2 [M+H]+.
The Boc protecting group in compound 6-A-24 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.57 (dd, J=4.7, 1.7 Hz, 1H), 7.96 (dd, J=7.9, 1.7 Hz, 1H), 7.61-7.49 (m, 4H), 7.44-7.27 (m, 5H), 7.23 (dd, J=7.9, 4.7 Hz, 1H), 5.82 (s, 2H), 3.79 (s, 2H), 3.66 (s, 2H), 2.97 (t, J=5.4 Hz, 2H), 2.83 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.40, 148.88, 148.02, 141.88, 139.78, 137.70, 131.91, 129.44, 129.22, 129.01, 128.76, 128.45, 127.84, 127.37, 125.23, 118.33, 115.99, 62.64, 51.65, 48.62, 43.75, 25.75; MS (ESI) m/z: 450.2 [M+H]+.
The Boc protecting group in compound 6-A-18 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.56 (dd, J=4.6, 1.6 Hz, 1H), 7.90 (dd, J=7.9, 1.6 Hz, 1H), 7.52-7.45 (m, 2H), 7.31-7.24 (m, 1H), 7.20-7.09 (m, 3H), 6.97-6.87 (m, 3H), 5.70 (s, 2H), 3.73 (s, 2H), 3.60 (s, 2H), 2.90 (t, J=5.3 Hz, 2H), 2.78 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 164.64, 163.58, 161.35, 148.84, 148.06, 140.71, 139.48, 133.70, 131.82, 130.74, 129.91, 127.64, 124.59, 118.18, 115.90, 115.58, 115.12, 114.84, 114.35, 114.07, 62.01, 51.59, 48.75, 43.33, 25.66; MS (ESI) m/z: 418.2 [M+H]+.
The Boc protecting group in compound 6-A-18 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.57 (dd, J=4.6, 1.6 Hz, 1H), 7.91 (dd, J=7.9, 1.7 Hz, 1H), 7.52-7.46 (m, 2H), 7.39 (s, 1H), 7.26-7.22 (m, 3H), 7.19 (dd, J=7.9, 4.7 Hz, 1H), 6.94-6.88 (m, 2H), 5.70 (s, 2H), 3.72 (s, 2H), 3.60 (s, 2H), 2.92 (t, J=5.4 Hz, 2H), 2.78 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, Chloroform-d) δ 163.59, 161.34, 160.34, 148.85, 148.08, 140.08, 139.44, 134.34, 133.67, 131.81, 130.72, 129.68, 129.01, 127.63, 127.52, 127.18, 118.17, 115.90, 115.13, 114.84, 61.98, 51.60, 48.74, 43.34, 25.66; MS (ESI) m/z: 434.1 [M+H]+.
The Boc protecting group in compound 6-A-18 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.56 (dd, J=4.6, 1.6 Hz, 1H), 7.90 (dd, J=7.9, 1.7 Hz, 1H), 7.55-7.46 (m, 3H), 7.40-7.36 (m, 1H), 7.31-7.27 (m, 1H), 7.21-7.15 (m, 2H), 6.94-6.88 (m, 2H), 5.70 (s, 2H), 3.70 (s, 2H), 3.59 (s, 2H), 2.91 (t, J=5.4 Hz, 2H), 2.77 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, Chloroform-d) δ 163.58, 161.33, 160.33, 148.85, 148.07, 140.41, 139.45, 133.64, 131.90, 131.82, 130.73, 130.63, 130.44, 129.99, 127.65, 122.63, 118.17, 115.88, 115.12, 114.84, 61.93, 51.59, 48.74, 43.34, 25.65; MS (ESI) m/z: 478.1 [M+H]+.
The Boc protecting group in compound 6-A-19 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.6 Hz, 1H), 7.92 (dd, J=7.9, 1.6 Hz, 1H), 7.44-7.39 (m, 2H), 7.29-7.24 (m, 1H), 7.21-7.09 (m, 5H), 6.99-6.92 (m, 1H), 5.70 (s, 2H), 3.74 (s, 2H), 3.60 (s, 2H), 2.92 (t, J=5.3 Hz, 2H), 2.78 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, Chloroform-d) δ 164.65, 161.34, 148.86, 148.05, 140.71, 139.53, 136.39, 132.88, 131.83, 130.46, 130.25, 129.90, 129.79, 128.34, 127.68, 124.58, 118.21, 115.92, 114.36, 114.08, 62.02, 51.60, 48.74, 43.45, 25.70; MS (ESI) m/z: 434.1 [M+H]+.
The Boc protecting group in compound 6-A-19 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.56 (dd, J=4.6, 1.6 Hz, 1H), 7.93 (dd, J=7.9, 1.7 Hz, 1H), 7.44-7.38 (m, 3H), 7.26-7.17 (m, 6H), 5.70 (s, 2H), 3.72 (s, 2H), 3.60 (s, 2H), 2.94 (t, J=5.5 Hz, 2H), 2.79 (t, J=5.7 Hz, 2H). 13C NMR (75 MHz, CDCl3) δ 161.34, 148.87, 148.06, 140.09, 139.51, 136.37, 134.35, 132.89, 131.83, 130.23, 129.68, 129.01, 128.35, 127.66, 127.52, 127.17, 118.21, 115.91, 61.98, 51.60, 48.73, 43.46, 25.69; MS (ESI) m/z: 450.1 [M+H]+.
The Boc protecting group in compound 6-A-19 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.5 Hz, 1H), 7.89 (dd, J=7.9, 1.6 Hz, 1H), 7.54 (s, 1H), 7.43-7.36 (m, 3H), 7.31-7.26 (m, 1H), 7.21-7.14 (m, 4H), 5.69 (s, 2H), 3.70 (s, 2H), 3.58 (s, 2H), 2.89 (t, J=5.2 Hz, 2H), 2.76 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, Chloroform-d) δ 161.31, 148.87, 148.02, 140.43, 139.54, 136.39, 132.87, 131.90, 131.84, 130.44, 130.26, 130.00, 128.34, 127.65, 127.60, 122.64, 118.23, 115.88, 61.93, 51.60, 48.74, 43.45, 25.68; MS (ESI) m/z: 494.1 [M+H]+.
The Boc protecting group in compound 6-A-20 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.6 Hz, 1H), 7.92 (dd, J=7.9, 1.6 Hz, 1H), 7.35 (s, 4H), 7.27-7.09 (m, 4H), 6.99-6.92 (m, 1H), 5.69 (s, 2H), 3.74 (s, 2H), 3.60 (s, 2H), 2.92 (t, J=5.3 Hz, 2H), 2.79 (t, J=5.6 Hz, 2H); 13C NMR (75 MHz, Chloroform-d) δ 164.65, 161.34, 148.86, 148.05, 140.69, 139.53, 136.89, 131.83, 131.30, 130.59, 129.90, 127.69, 124.57, 124.54, 121.06, 118.22, 115.92, 115.58, 114.37, 114.09, 62.00, 51.60, 48.73, 43.51, 25.70; MS (ESI) m/z: 478.1 [M+H]+.
The Boc protecting group in compound 6-A-20 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.55 (dd, J=4.6, 1.6 Hz, 1H), 7.92 (dd, J=7.9, 1.6 Hz, 1H), 7.40-7.37 (m, 1H), 7.35 (s, 4H), 7.26-7.23 (m, 3H), 7.19 (dd, J=7.9, 4.7 Hz, 1H), 5.68 (s, 2H), 3.72 (s, 2H), 3.59 (s, 2H), 2.92 (t, J=5.3 Hz, 2H), 2.78 (t, J=5.6 Hz, 2H). 13C NMR (75 MHz, Chloroform-d) δ 161.33, 148.87, 148.03, 140.09, 139.54, 136.89, 134.35, 131.84, 131.30, 130.59, 129.68, 129.01, 127.63, 127.52, 127.18, 121.06, 118.23, 115.90, 61.98, 51.59, 48.73, 43.51, 25.69; MS (ESI) m/z: 494.1 [M+H]+.
The Boc protecting group in compound 6-A-20 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-bromobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. 1H NMR (300 MHz, Chloroform-d) δ 8.54 (dd, J=4.6, 1.6 Hz, 1H), 7.89 (dd, J=7.9, 1.6 Hz, 1H), 7.54 (s, 1H), 7.41-7.28 (m, 6H), 7.20-7.14 (m, 2H), 5.67 (s, 2H), 3.70 (s, 2H), 3.58 (s, 2H), 2.90 (t, J=5.3 Hz, 2H), 2.76 (t, J=5.5 Hz, 2H); 13C NMR (75 MHz, Chloroform-d) δ 161.30, 148.89, 148.00, 140.39, 139.57, 136.90, 131.90, 131.86, 131.30, 130.60, 130.45, 130.01, 127.67, 127.56, 122.64, 121.06, 118.26, 115.87, 61.92, 51.58, 48.74, 43.52, 25.67; MS (ESI) m/z: 538.0 [M+H]+.
The Boc protecting group in compound 6-B-1 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 449.2 [M+H]+.
The Boc protecting group in compound 6-C-1 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 451.2 [M+H]+.
The compound was prepared by reacting compound 7-1 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 407.3 [M+H]+.
The Boc protecting group in compound 6-A-24 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 475.3 [M+H]+.
The Boc protecting group in compound 6-A-18 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 425.3 [M+H]+.
The Boc protecting group in compound 6-A-19 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 441.3 [M+H]+.
The Boc protecting group in compound 6-A-20 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 485.2 [M+H]+.
The Boc protecting group in compound 6-A-23 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 437.3 [M+H]+.
The Boc protecting group in compound 6-A-37 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 452.3 [M+H]+.
100 mg of I-88 was dissolved in methanol, and 0.1 mL of aqueous formaldehyde solution was added, followed by the addition of 20 mg of Pd—C (10%) under nitrogen atmosphere. The mixture was stirred under hydrogen atmosphere (1 atm) for 3 h. The reaction mixture was filtered through celite to remove the catalyst. The mixture containing I-89 and I-90 which remained after concentration of the filtrate was separated by semi-preparative HPLC to give pure compounds I-89 and 1-90. I-89: MS (ESI) m/z: 436.3 [M+H]+; I-90: MS (ESI) m/z: 450.3 [M+H]+.
The Boc protecting group in compound 6-A-21 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 421.3 [M+H]+.
The Boc protecting group in compound 6-A-21 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 430.3 [M+H]+.
The Boc protecting group in compound 6-A-21 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 414.3 [M+H]+.
The Boc protecting group in compound 6-A-22 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 435.3 [M+H]+.
The Boc protecting group in compound 6-A-22 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 428.3 [M+H]+.
The Boc protecting group in compound 6-A-22 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 444.3 [M+H]+.
The Boc protecting group in compound 6-A-38 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 474.3 [M+H]+.
The Boc protecting group in compound 6-A-39 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 451.3 [M+H]+.
The Boc protecting group in compound 6-A-23 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 430.3 [M+H]+.
The Boc protecting group in compound 6-A-23 was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 446.3 [M+H]+.
The Boc protecting group in compound 6-D was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 439.3 [M+H]+.
The Boc protecting group in compound 6-E was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 455.3 [M+H]+.
The Boc protecting group in compound 6-F was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 455.3 [M+H]+.
The Boc protecting group in compound 6-G was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 455.3 [M+H]+.
The Boc protecting group in compound 6-H was first removed by referring to the method in Example 58, and then the compound was prepared by reacting the resulting amine with benzyl bromide by referring to the synthetic method for compound I-1 in Example 59. MS (ESI) m/z: 456.3 [M+H]+.
0.22 g (2 mmol) of 2-methylbenzylamine was dissolved in 10 mL of 1,4-dioxane, and 0.49 g of 2-methylthio-imidazoline hydroiodide (compound 8, 2 mmol) was added to the above solution. The mixture was heated to 70° C. and reacted for 2 h under argon atmosphere. The reaction mixture was concentrated by rotary evaporation to dryness to remove the solvent to give compound 9-1, which was directly used in the next step without purification.
The compound 9-1 obtained in the above step was dissolved in 10 mL of methanol, and 0.54 g of ethyl 1-N-tert-butoxycarbonyl-4-piperidone-3-carboxylate (compound 2, 2 mmol) and 0.16 g of sodium methoxide (3 mmol) were added to the above solution. The reaction solution was heated at reflux for 2 h. After the reaction was completed as detected by TLC, the reaction mixture was concentrated by rotary evaporation to remove the solvent, and 10 mL of water was added to the residue. The mixture was adjusted to pH 7 with 1 N hydrochloric acid and extracted with ethyl acetate (15 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated, and the residue was purified by column chromatography to give compound 10-1 (0.44 g, yield over two steps: 56%), MS (ESI) m/z: 397.2 [M+H]+.
0.4 g (1 mmol) of compound 10-1 was dissolved in 3 mL of DMSO, and 0.84 g of 2-iodoxybenzoic acid (IBX, 3 mmol) was added to the above solution. The mixture was reacted at room temperature for 6 h. After the reaction was completed as detected by TLC, 20 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated, and the residue was purified by column chromatography to give compound 11-1 (0.23 g, yield: 58%), MS (ESI) m/z: 395.2 [M+H]+, HRMS (ESI) m/z: [M+H]+ 395.2004, calcd. 395.2005.
0.2 g of compound 11-1 was dissolved in 3 mL of methanol, and 1 mL of a saturated solution of HCl in methanol was added. The mixture was stirred at room temperature for 12 h. The reaction mixture was concentrated by rotary evaporation to dryness to remove the solvent. 10 mL of water was added to the residue, and the mixture was neutralized with a saturated NaHCO3 solution to pH 8 and extracted with dichloromethane (15 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give compound 12-1 (0.15 g, yield: 91%), MS (ESI) m/z: 295.2 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with benzylamine as a starting material, MS (ESI) m/z: 281.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-fluorobenzylamine as a starting material, MS (ESI) m/z: 299.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 2,4-difluorobenzylamine as a starting material, MS (ESI) m/z: 317.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-chlorobenzylamine as a starting material, MS (ESI) m/z: 315.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-methylbenzylamine as a starting material, MS (ESI) m/z: 295.3 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-ethylbenzylamine as a starting material, MS (ESI) m/z: 309.3 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-trifluoromethylbenzylamine as a starting material, MS (ESI) m/z: 349.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 12-1 in Example 163 with 4-methoxybenzylamine as a starting material, MS (ESI) m/z: 311.3 [M+H]+.
100 mg (0.34 mmol) of compound 12-1 was dissolved in 3 mL of DMF, and 60 mg of benzyl bromide (0.35 mmol) and 70 mg of potassium carbonate (0.5 mmol) were separately added to the above solution. The mixture was reacted at room temperature for 6 h. After the reaction was completed as detected by TLC, 15 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was first separated by column chromatography and then further purified by HPLC (eluent: acetonitrile containing 0.1% TFA and water) to give a trifluoroacetate salt of the target compound I-82 (88 mg, yield: 52%). MS (ESI) m/z: 385.2 [M+H]+.
The compound was prepared by reacting compound 12-2 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 371.2 [M+H]+.
The compound was prepared by reacting compound 12-2 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 389.2 [M+H]+.
The compound was prepared by reacting compound 12-2 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 405.1 [M+H]+.
The compound was prepared by reacting compound 12-2 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 385.2 [M+H]+.
The compound was prepared by reacting compound 12-2 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared by reacting compound 12-3 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 389.2 [M+H]+.
The compound was prepared by reacting compound 12-3 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 407.2 [M+H]+.
The compound was prepared by reacting compound 12-3 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 423.1 [M+H]+.
The compound was prepared by reacting compound 12-3 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 403.2 [M+H]+.
The compound was prepared by reacting compound 12-3 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 414.2 [M+H]+.
The compound was prepared by reacting compound 12-5 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 405.2 [M+H]+.
The compound was prepared by reacting compound 12-5 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 423.2 [M+H]+.
The compound was prepared by reacting compound 12-5 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 439.1 [M+H]+.
The compound was prepared by reacting compound 12-5 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 419.2 [M+H]+.
The compound was prepared by reacting compound 12-5 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 430.2 [M+H]+.
The compound was prepared by reacting compound 12-4 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 407.2 [M+H]+.
The compound was prepared by reacting compound 12-4 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 425.2 [M+H]+.
The compound was prepared by reacting compound 12-4 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 441.1 [M+H]+.
The compound was prepared by reacting compound 12-4 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 421.2 [M+H]+.
Compound I-126 was prepared by reacting compound 12-4 with 3-cyanobenzyl bromide by the same method as that for compound I-106 in Example 172, MS (ESI) m/z: 432.2 [M+H]+.
The compound was prepared by reacting compound 12-8 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 439.2 [M+H]+.
The compound was prepared by reacting compound 12-8 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 457.2 [M+H]+.
The compound was prepared by reacting compound 12-8 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 473.2 [M+H]+.
The compound was prepared by reacting compound 12-8 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 453.2 [M+H]+.
The compound was prepared by reacting compound 12-8 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 464.2 [M+H]+.
The compound was prepared by reacting compound 12-6 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 385.2 [M+H]+.
The compound was prepared by reacting compound 12-6 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 403.2 [M+H]+.
The compound was prepared by reacting compound 12-6 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 419.2 [M+H]+.
The compound was prepared by reacting compound 12-6 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 399.2 [M+H]+.
The compound was prepared by reacting compound 12-6 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 410.2 [M+H]+.
The compound was prepared by reacting compound 12-7 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 399.2 [M+H]+.
The compound was prepared by reacting compound 12-7 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 417.2 [M+H]+.
The compound was prepared by reacting compound 12-7 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 433.2 [M+H]+.
The compound was prepared by reacting compound 12-7 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 413.2 [M+H]+.
The compound was prepared by reacting compound 12-7 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 424.2 [M+H]+.
The compound was prepared by reacting compound 12-9 with benzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 401.2 [M+H]+.
The compound was prepared by reacting compound 12-9 with 3-fluorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 419.2 [M+H]+.
The compound was prepared by reacting compound 12-9 with 3-chlorobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 435.2 [M+H]+.
The compound was prepared by reacting compound 12-9 with 3-methylbenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 415.2 [M+H]+.
The compound was prepared by reacting compound 12-9 with 3-cyanobenzyl bromide by referring to the synthetic method for compound I-106 in Example 172, MS (ESI) m/z: 426.2 [M+H]+.
0.63 g of compound 13 (4 mmol) was dissolved in 10 mL of dichloromethane, and 2 mL of oxalyl chloride and two drops of DMF were added to the solution at 0° C. under stirring. The reaction solution was warmed to room temperature, stirred for half an hour, and concentrated by rotary evaporation under reduced pressure to remove the solvent and excessive oxalyl chloride. The resulting acyl chloride was re-dissolved in 5 mL of dichloromethane for later use. 0.73 g of compound 14-1 was dissolved in 10 mL of dichloromethane, and 1.5 mL of Et3N was added. The solution was cooled to 0° C., and an acyl chloride solution prepared from compound 13 was slowly added to the above solution under stirring. The mixture was stirred at room temperature for 3 h, and concentrated by rotary evaporation to remove the solvent, and the residue was separated by column chromatography to give compound 15-1, MS (ESI) m/z: 323.1 [M+H]+.
Compound 15-1 obtained in the above step was dissolved in 5 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added at 0° C. under stirring. The reaction solution was warmed to room temperature, stirred for 1 h, and concentrated by rotary evaporation to remove the solvent. The residue was dissolved in 15 mL of 1,4-dioxane, and 0.4 g of NaH (purity: 60%) was added to the above solution in portions, heated to 80° C., and stirred overnight under nitrogen atmosphere. 10 mL of methanol was added to the above reaction solution to destroy excess sodium hydride, and then the mixture was concentrated by rotary evaporation to remove the solvent. The residue was purified by column chromatography to give compound 16-1, MS (ESI) m/z: 187.1 [M+H]+.
Compound 16-1 obtained in the above reaction was dissolved in 15 mL of methanol, and 0.5 mL of concentrated hydrochloric acid was added, followed by the addition of 50 mg of 10% Pd—C under nitrogen atmosphere. The mixture was stirred overnight under hydrogen atmosphere and filtered through celite to remove the solid. The filtrate was concentrated to give a hydrochloride salt of compound 17-1, MS (ESI) m/z: 191.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 17-1 in Example 212 with compound 13 and compound 14-2 as starting materials, MS (ESI) m/z: 192.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 17-1 in Example 212 with compound 13 and compound 14-3 as starting materials, MS (ESI) m/z: 191.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 17-1 in Example 212 with compound 13 and compound 14-4 as starting materials, MS (ESI) m/z: 191.1 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 17-1 in Example 212 with compound 13 and compound 14-5 as starting materials, MS (ESI) m/z: 190.1 [M+H]+.
0.63 g of compound 13 (4 mmol) was dissolved in 10 mL of dichloromethane, and 2 mL of oxalyl chloride and two drops of DMF were added to the above solution under stirring at 0° C. The mixture was warmed to room temperature, stirred for half an hour, and concentrated by rotary evaporation under reduced pressure to remove the solvent and excess oxalyl chloride. The resulting acyl chloride was re-dissolved in 5 mL of dichloromethane, and the above acyl chloride solution was slowly added to 10 mL of dichloromethane solution containing 0.67 g of 2,4-dimethoxybenzylamine and 1.5 mL of triethylamine which cooled in an ice bath. The reaction solution was warmed to room temperature, stirred for 3 h, and concentrated by rotary evaporation to remove the solvent. The residue was separated by column chromatography to give compound 20, MS (ESI) m/z: 307.1 [M+H]+.
Compound 20 obtained in the above step was dissolved in 15 mL of DMF, and 0.5 g of imidazole and 0.4 g of NaH (purity: 60%) were separately added to the solution. The mixture was heated to 100° C., and stirred overnight under nitrogen atmosphere. After cooling to room temperature, 30 mL of water was added to the above reaction solution, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined, washed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to remove the solvent. The residue was purified by column chromatography to give compound 22-1, MS (ESI) m/z: 339.2 [M+H]+.
Compound 22-1 obtained in the above step was dissolved in 15 mL of DMF, and 0.1 g of CuI, 0.5 g of potassium tert-butoxide, and 0.2 g of 1,10-phenanthroline were separately added to the solution. The resulting mixture was reacted at 120° C. overnight under oxygen atmosphere. After the reaction was completed, the reaction mixture was cooled to room temperature. 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined, washed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to remove the solvent. The residue was purified by column chromatography to give compound 23-1, MS (ESI) m/z: 337.2 [M+H]+.
Compound 23-1 obtained in the above reaction was dissolved in 15 mL of methanol, and 0.5 mL of concentrated hydrochloric acid was added, followed by the addition of 50 mg of 10% Pd—C under nitrogen atmosphere. The mixture was stirred overnight under hydrogen atmosphere and filtered through celite to remove the solid. The filtrate was concentrated to give a hydrochloride salt of compound 24-1, MS (ESI) m/z: 341.2 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 24-1 in Example 217 with compound 20 and 1,2,4-triazole as starting materials, MS (ESI) m/z: 342.2 [M+H]+.
The compound was prepared by referring to the synthetic method for compound 24-1 in Example 217 with compound 20 and pyrazole as starting materials, MS (ESI) m/z: 341.2 [M+H]+.
190 mg of compound 17-1 was dissolved in 10 mL of methanol, and 140 mg of m-chlorobenzaldehyde and 120 mg of NaBH3CN were separately added to the solution, followed by the addition of a drop of concentrated sulfuric acid. The mixture was reacted at room temperature for 3 h. 1 mL of water was added, and the mixture was concentrated by rotary evaporation to remove the solvent. 15 mL of water was added to the residue, and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation to remove the solvent. The residue was dissolved in 5 mL of DMF, 0.2 g of 5-bromomethylbenzo[1,3]dioxolane and 0.2 g of potassium carbonate were separately added, and the mixture was stirred at room temperature overnight. 10 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to remove the solvent. The residue was first purified by silica gel column chromatography and then further purified by semi-preparative HPLC to give compound I-147. MS (ESI) m/z: 449.2 [M+H]+.
170 mg of compound 24-1 was dissolved in 5 mL of DMF, and 120 mg of 3-chlorobenzyl bromide and 0.2 mL of triethylamine were separately added to the solution. The mixture was stirred at room temperature overnight. 5 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography, and the resulting product was dissolved in 2 mL of dichloromethane. 10 mL of trifluoroacetic acid was added to the solution, and the mixture was stirred at room temperature for 2 h and concentrated by rotary evaporation to remove the solvent and excess trifluoroacetic acid. The residue was dissolved in 5 mL of DMF, and 120 mg of 6-bromomethyl[2,3]dihydrobenzo[1,4]dioxin and 0.2 g of potassium carbonate were separately added to the solution. The mixture was stirred at room temperature overnight, 10 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to remove the solvent. The residue was first purified by silica gel column chromatography and then further purified by semi-preparative HPLC to give compound I-148. MS (ESI) m/z: 463.2 [M+H]+.
The compound was prepared from compound 17-2 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 440.2 [M+H]+.
The compound was prepared from compound 17-2 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 474.2 [M+H]+.
The compound was prepared from compound 17-2 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 465.2 [M+H]+.
The compound was prepared from compound 24-2 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 406.2 [M+H]+.
The compound was prepared from compound 24-2 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 424.2 [M+H]+.
The compound was prepared from compound 24-2 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 440.1 [M+H]+.
The compound was prepared from compound 24-2 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 431.2 [M+H]+.
The compound was prepared from compound 17-3 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 405.2 [M+H]+.
The compound was prepared from compound 17-3 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 396.2 [M+H]+.
The compound was prepared from compound 17-3 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 385.2 [M+H]+.
The compound was prepared from compound 24-3 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 439.2 [M+H]+.
The compound was prepared from compound 24-3 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 464.2 [M+H]+.
The compound was prepared from compound 24-3 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 405.2 [M+H]+.
The compound was prepared from compound 24-3 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 439.1 [M+H]+.
The compound was prepared from compound 24-3 by referring to the synthetic method for compound I-148 in Example 221, MS (ESI) m/z: 430.2 [M+H]+.
The compound was prepared from compound 17-4 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 439.2 [M+H]+.
The compound was prepared from compound 17-4 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 464.2 [M+H]+.
The compound was prepared from compound 17-4 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 439.1 [M+H]+.
The compound was prepared from compound 17-4 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 430.2 [M+H]+.
The compound was prepared from compound 17-5 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 438.1 [M+H]+.
The compound was prepared from compound 17-5 by referring to the synthetic method for compound I-147 in Example 220, MS (ESI) m/z: 438.2 [M+H]+.
The EC50 values for activating ClpP hydrolysis of FITC-casein and the activity for inhibiting the growth of MV4; 11 cells of the compounds of formula I as ClpP agonists were determined by the methods as described in the literature (Cancer Cell 2019, 35, 721-737). The specific procedures were as follows:
Test compounds were each prepared into a 100 mM stock solution in DMSO and stored. and 1 μL of stock solution of each of the compounds was taken and diluted to 10 mM by adding 9 μL of DMSO for later use.
The compounds of formula I of the present invention were each diluted at concentrations of 200 μM, 50 μM, 10 μM, 2 μM, 500 nM, 100 nM, 20 nM, and 2 nM in an assay buffer. 50 μL of each of the compounds at different concentrations was added to each well, with 2 replicate wells. The final concentrations were 100 μM, 25 μM, 5 μM, 1 μM, 250 nM, 50 nM, 10 nM, and 1 nM.
The protein hClpP purified by referring to the method in the literature (Cancer Cell 2019, 35, 721-737.) was at a concentration of 14 mg/mL, which was converted to a concentration of 1000 μM. The protein was diluted to 4 μM in an assay buffer. 25 μL of protein was added to each well, and incubated at 37° C. for 30 min. The final concentration was 1 μM.
A FITC-Casein stock solution labeled by referring to the method in the literature (Cancer Cell 2019, 35, 721-737) was at 200 mM, which was diluted to 16 μM in an assay buffer. 25 μL of FITC-Casein was added to each well. The final concentration was 4 μM. The mixture was immediately loaded on the machine for detection.
FITC-Casein and FITC-Casein+ClpP were used as negative controls. The positive drugs ONC201 and ONC212 were used as positive control groups, and the positive drugs ONC201 and ONC212 were synthesized by the method in the literature (Angewandte Chemie International Edition 2014, 53, 6628-6631).
The detection temperature was 37° C. The plate was shaken for 3 s. The excitation light was at the wavelength of 485±20 nm, and the emission light was at the wavelength of 525±20 nm. Kinetic parameters were set, and scanning was performed once every 1 min for 15 min.
The results for the EC50 values for activating ClpP hydrolysis of FITC-casein and the activity for inhibiting the growth of MV4; 11 cells of the compounds of formula I as ClpP agonists were shown in the table below.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202110805091.6 | Jul 2021 | CN | national |
| 202210814872.6 | Jul 2022 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/105579 | 7/14/2022 | WO |
