Patent Application
20250042920
The invention belongs to the field of medicinal chemistry, in particular to Imidazothiazole derivatives and a preparation method and application thereof.
Metabolic syndrome is a group of clinical syndromes determined by genetic and environmental factors and characterized by the co-occurrence of multiple metabolic diseases such as obesity and type 2 diabetes mellitus (T2D), which can further lead to dyslipidemia, hypertension, non-alcoholic fatty liver disease, and gout. With the development of social economy and the change of people's lifestyle (increase in energy intake and decrease in exercise, etc.), the incidence of metabolic syndrome has shown an increasing trend globally year by year, and has become a global public health challenge.
Nonalcoholic fatty liver disease (NAFLD) is a genetic, environmental, metabolic and stress related liver disease characterized by hepatic parenchymal cell steatosis and fat storage. Clinically, there are three types: simple fatty liver, steatohepatitis (NASH) and fatty cirrhosis.
MNK (Human mitogenic protein kinase interacting enzyme), including two subtypes MNK1 and MNK2. MNK has been shown to phosphorylate elF4E in vivo, thereby regulating protein synthesis in organisms. Previous studies have found that regulating MNK activity can regulate body weight, glucose tolerance, enhance insulin sensitivity, energy consumption capacity, liver fat accumulation and inflammation in animal fat, suggesting that MNK can be used as a potential drug development target for metabolic diseases such as diabetes, obesity and NAFLD.
There is an urgent need for more types of small molecule compounds that can better inhibit MNK activity in this field.
In order to solve the above technical problems, we have invented a series of imidazothiazole or imidazothiadiazole derivative compounds, which can regulate blood sugar in the body, reduce weight gain, reduce fat accumulation and other changes by regulating the activity of MNK protease, thereby improving the condition of patients with diabetes and other metabolic diseases.
The present invention provides an imidazothiazole derivative, a stereoisomer, a tautomer, a geometric isomer, or a pharmaceutically acceptable salt thereof, wherein the imidazothiazole derivative has a structure of formula (I):
As described in the present invention, the C1-C6 alkyl refers to a straight or branched chain saturated monovalent hydrocarbon group containing 1, 2, 3, 4, 5 or 6 carbon atoms, and representative examples include but are not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, etc. C1-C6 alkyl can further preferably be a C1-C3 alkyl.
As described in the present invention, the C1-C6 haloalkyl refers to one or more hydrogen atoms in the “C1-C6 alkyl” as defined above is substituted with the same or different halogen atoms. C1-C6 haloalkyl can further preferably be a C1-C3 haloalkyl. Representative examples include, but are not limited to, trifluoromethyl, difluoromethyl, monofluoromethyl, trichloromethyl, 1,1,1-trifluoroethyl, pentafluoroethyl, etc.
As described in the present invention, the C1-C6 alkylamino refers to one or two hydrogen atoms in amino (—NH2) are substituted with the same or different above-defined “C1-C6 alkyl”; i.e., it can be referred as —NR1R2, wherein R1 and R2 are each independently selected from H and C1-C6 alkyl, and R1 and R2 cannot be H at the same time.
As described in the present invention, the C1-C6 alkoxy refers to —OR3, wherein R3 is selected from C1-C6 alkyl; C1-C6 alkoxy can further preferably be C1-C3 alkoxy, and more further preferably be methoxy and ethoxy.
As described in the present invention, the C1-C6 alkoxycarbonyl refers to —C(O)R4, wherein R4 is selected from C1-C6 alkoxy.
As described in the present invention, the 5 to 6-membered heterocyclic ring refers to a ring system containing ring carbon atoms and 1 to 4 ring heteroatoms (preferably 1, 2 or 3 ring heteroatoms), wherein each ring heteroatom is independently selected from nitrogen, oxygen and sulfur; in heterocyclic groups containing one or more nitrogen atoms, the attachment site can be either carbon or nitrogen atoms, as long as the valence allows. Further preferred is piperazine ring, morpholine ring, piperidine ring, hexahydropyran ring, tetrahydrofuran ring, tetrahydrothiophene ring, pyrrole ring, tetrahydropyrrole ring and so on.
As described in the present invention, the aryl is preferably a monocyclic or polycyclic aryl containing 6-12 carbon atoms, preferably is phenyl, naphthalenyl, etc.; heteraryl is preferably 5 to 6-membered heteraryl, 5 to 6-membered heteraryl refers to a heteraryl system containg ring carbon atoms and 1 to 4 ring heteroatoms (preferably 1, 2, or 3 ring heteroatoms), wherein each ring heteroatom is independently selected from nitrogen, oxygen, sulfur. Further preferred is furanyl, thiophenyl, pyridinyl, thiazolyl, imidazolyl and so on.
As described in the present invention, the halogen atom or halogen is preferably fluorine, chlorine, bromine or iodine.
The above-mentioned imidazothiazole derivative having a structure of formula (I) is preferably compounds 1 to 55 or a stereoisomer, tautomer, geometric isomer or a pharmaceutically acceptable salt thereof,
Another embodiment of the present invention provides a method for preparing the above imidazothiazole derivative having the structure of formula (I), wherein comprising the following steps:
under alkaline conditions, thereby obtaining the compound of general formula (2), wherein R3 and R4 are defined as above.
Another embodiment of the present invention provides a method for preparing the above imidazothiazole derivative having the structure of formula (I), wherein comprising the following steps:
wherein R1 is defined as above, X is halogen, preferably chlorine, bromine, or iodine;
wherein R3 is defined as above.
Another embodiment of the present invention provides a method for preparing the above imidazothiazole derivative having the structure of formula (I), wherein the imidazothiazole derivative having the structure of formula (I) is selected from general formula (1) wherein R2 is 4-cyanophenyl, and is represented by compound N, comprising the following steps:
making compound M undergo a condensation reaction with R1H, thereby obtaining compound N, and the definition of R1 is the same as above.
Another embodiment of the present invention provides an intermediate for the preparation of the imidazothiazole derivative having the structure of formula (I), wherein the intermediate has a structure of formula (II)
wherein the definition of R1 is the same as above, and X is halogen, preferably is chlorine, bromine or iodine.
Another embodiment of the present invention provides an intermediate for the preparation of the imidazothiazole derivative having the structure of formula (I), wherein the intermediate has a structure of formula (III)
wherein the definition of R3 is the same as above.
Another embodiment of the present invention provides an intermediate for the preparation of formula (II) wherein the intermediate has a structure of formula (IV):
wherein X is a halogen, preferably is chlorine, bromine or iodine.
Another embodiment of the invention provides an intermediate for the preparation of formula (III) wherein the intermediate has a structure of formula (V):
Another embodiment of the present invention provides an intermediate for the preparation of the imidazothiazole derivative having the structure of formula (I), wherein the intermediate has the following structure:
Another embodiment of the present invention provides a use of compounds of formula (II), formula (III), formula (IV), and formula (V) in preparing the imidazothiazole derivative having the structure of formula (I).
Another embodiment of the present invention provides a use of the imidazothiazole derivative having the structure of formula (I), the stereoisomer, tautomer, or geometric isomer thereof, or the pharmaceutically acceptable salt thereof in inhibiting kinase activity of MNK1 or MNK2 or variants thereof; or in the preparation of a drug for preventing and/or treating cancers caused by abnormal levels of MNK1 and/or MNK2.
Another embodiment of the present invention provides a use of the imidazothiazole derivative having the structure of formula (I), the stereoisomer, tautomer, or geometric isomer thereof, or the pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and/or treating metabolic diseases associated with MNK activity. The metabolic diseases associated with MNK activity are selected from type 1 diabetes mellitus, type 2 diabetes mellitus, hyperlipidemia, obesity, fatty liver disease, and complications thereof and related disorders thereof.
Another embodiment of the present invention provides a use of the imidazothiazole derivative having the structure of formula (I), the stereoisomer, tautomer, or geometric isomer thereof, or the pharmaceutically acceptable salt thereof in the preparation of MNK1 and/or MNK2 inhibitors.
Another embodiment of the present invention provides a pharmaceutical composition wherein the pharmaceutical composition use the imidazothiazole derivative having the structure of formula (I), the stereoisomer, the tautomer, or the geometric isomer thereof, or the pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition may also comprise pharmaceutically acceptable excipients. The pharmaceutical composition may also include other MNK1 and/or MNK2 inhibitors (marketed therapeutics). The dosage form thereof can be solid preparation, liquid preparation or semi-solid preparation, preferably tablet, capsule, injection and so on.
Methods and techniques of the present invention are generally implemented according to traditional methods known in the art, unless otherwise indicated. The terms described in the invention are named according to chemistry, biology and pharmacology, and the experimental methods and techniques are known and commonly used in the field. Standard techniques are used in chemosynthesis, chemical analysis, preparations and formulations for medicine, and the treatment for patients. Unless otherwise stated, the scientific and technical terms used in the present invention shall have those meanings commonly understood by those skilled in the art. While the following terms have the following definitions:
Unless otherwise specified, all compounds present in the present invention are intended to include all possible optical isomers, such as single chiral compounds or mixtures of various chiral compounds (i.e., racemes). Among all compounds of the present invention, each chiral carbon atom may optionally be in R configuration or S configuration, or a mixture of the two configurations.
Diabetes mellitus and its complications are specifically described as impaired glucose tolerance, diabetic gangrene, diabetic arthropathy, diabetic osteopenia, diabetic glomerulosclerosis, diabetic nephropathy, diabetic dermatopathy, diabetic neuropathy, diabetic cataract, diabetic retinopathy, diabetic macular degeneration, diabetic foot syndrome, diabetic coma, diabetic hyperosmolar coma, hypoglycemic coma, hyperglycemic coma, diabetic acidosis, diabetic ketoacidosis, intracapillary glomerular nephropathy, diabetic muscular atrophy, diabetic autonomic neuropathy, diabetic mononeuropathy, diabetic polyneuropathy, diabetic vascular disease, diabetic peripheral vascular disease, diabetic ulcer, diabetic arthropathy, diabetic obesity.
Hyperlipidemia and its complications are: hypercholesterolemia, familial hypercholesterolemia, Verde's hyperlipoproteinemia, hyperβ-lipoproteinemia, hyperlipidemia, low density lipoproteinemia, pure hypertriglyceridemia, endogenous hypertriglyceridemia, simple hypercholesterolemia, simple hypertriglyceridemia, cardiovascular disease. Wherein cardiovascular diseases include: hypertension, ischemia, varicose veins, retinal vein occlusion, atherosclerosis, angina pectoris, myocardial infarction, stenocardia, pulmonary hypertension, congestive heart failure, glomerular disease, tubular interstitial disorders of the kidney, renal failure, vascular stenosis or cerebrovascular disease (stroke).
Fatty liver disease includes, but is not limited to, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and the resulting chronic inflammation leading to progressive fibrosis, cirrhosis, etc.
Using commercially available 5-bromo-2-amino-1, 3, 4-thiadiazole as raw material, chloracetaldehyde or 2-bromo-1, 1-diethoxyethane was reflow in alcohol solution (such as ethanol, n-butanol, etc.) for 1-2 days to obtain cycloidal product A, and then underwent Suzuki condensation reaction with 4-methyl phenylboronic acid to obtain compound B. C is obtained by nucleophilic substitution reaction with NBS (or NIS) in dichloromethane solution, and is hydrolyzed under alkaline conditions (e.g., in lithium hydroxide, sodium hydroxide, or potassium hydroxide aqueous solution, which may be a mixture of water, tetrahydrofuran, and alcohols), and then adjusted to acid (acid can be hydrochloric acid, sulfuric acid, acetic acid, hydrochloric acid). The carboxylic acid structure of compound D is obtained, and then amidation with amino compound R1H under the condition of amide condensation reagent (such as EDCI and NHS, EDCI and Hobt or EDCI and Hoat combination) is performed to obtain compound E. Finally, the Suzuki condensation reaction with the boronic acid compound B(OH)2R2 results in the final compound F (general formula 1), where R1 and R2 are defined as before.
The intermediate C obtained from the above reaction undergoes Suzuki condensation reaction with 4-cyanophenylboronic acid to obtain compound L, which is hydrolyzed under alkaline conditions (for example, in an aqueous solution of lithium hydroxide, sodium hydroxide, or potassium hydroxide, the solution can be a mixture of water, tetrahydrofuran, and alcohol), and then adjusted to acid (acid can be hydrochloric acid, sulfuric acid, acetic acid, hydrochloric acid). Aqueous solution such as formic acid) to obtain carboxylic acid structure compound M, and then under the condition of condensation reagent (such as EDCI and NHS, EDCI and Hobt or EDCI and Hoat combination) and R1H (amine compound, alcohol compound, sulfhydryl compound) condensation reaction to obtain the final compound N (R2 is the general formula 1 of 4-cyanophenyl). R1 is defined as above.
The following are detailed descriptions of the invention through specific preparation examples and embodiments, but the use and purpose of these exemplary embodiments are only used to illustrate the invention, and do not constitute any limitation of the actual scope of protection of the invention in any form, let alone limit the scope of protection of the invention to this.
compound A was obtained by reflux reaction of 5-bromo-2-amino-1,3, 4-thiadiazole with chloracetaldehyde in ethanol with a yield of 17.6%, 1H NMR (500 MHz, CDCl3) δ 7.76 (s, 1H), 7.36 (s, 1H).
2-bromoimidazo[2,1-b][1,3,4]thiadiazole (compound A) 300 mg (1.47 mmol) and methyl 4-formate phenylboronic acid 317 mg (1.76 mmol) under the action of catalyst, Silica gel column chromatography (PE:EA=10:1) was used to obtain a white solid of 93.2 mg (compound B) with a yield of 24.3% after Suzuki condensation reaction. 1H NMR (500 MHz, CDCl3) δ 8.19-8.15 (m, 2H), 7.97-7.93 (m, 2H), 7.81 (d, J=1.4 Hz, 1H), 7.37 (d, J=1.4 Hz, 1H), 3.97 (s, 3H).
After the reaction of 4-(imidazole [2,1-b][1,3,4]thiadiazole) methyl benzoate 350 mg (1.35 mmol) with 286 mg (1.62 mmol) n-bromosuccinimide, 340 mg (compound C) of light green solid was obtained by silica gel column chromatography, with a yield of 74.7%. 1H NMR (500 MHz, CDCl3)(8.18 (d, J=8.4 Hz, 2H), 7.99 (d, J=8.4 Hz, 2H), 7.29 (s, 1H), 3.98 (s, 3H). 13C NMR (125 MHz, CDCl3) δ 165.9, 161.5, 144.2, 133.8, 133.7, 133.1, 130.5, 126.9, 96.3, 52.5.
Take 4-(5-bromoimidazo[2,1-b][1,3,4]thiadiazole-2-methyl benzoate 340 mg (1 mmol), 424 mg (10 mmol) lithium hydroxide monohydrate mixed in 30 mL of THF/H2O (V:V=1:1) solution, At the end of the reaction, 320 mg of white solid was purified, that is, compound D, and the yield was 98%.
Methyl 4-acetylbenzoate (5 g, 28 mmol) was dissolved in acetonitrile (100 mL), then added to NBS (6.0 g, 33.6 mmol) and TsOH (482 mg, 2.8 mmol), and purified by column chromatography (PE:EA=10:1). The white solid was 4.6 g (i.e., compound G), and the yield was 63.9%. 1H NMR (400 MHz, CDCl3)(8.13 (d, J=8.1 Hz, 2H), 8.02 (d, J=8.1 Hz, 2H), 4.46 (s, 2H), 3.94 (s, 3H); 13C NMR (100 MHz, CDCl3) δ190.84, 165.95, 137.16, 134.62, 130.01, 128.87, 52.58, 30.71.
4-(2-bromoacetyl) methyl benzoate (4 g, 15.6 mmol), 2-amino-1,3, 4-thiadiazole (1.6 g, 16 mmol) and ethanol (100 mL) were added into 250 mL bottle of eggplant, and the reaction was carried out at 80° C. for 6 h under magnetic stirring. TLC detection of raw material reaction is complete. The reaction bottle was cooled to room temperature, filtered, and washed with ethanol for 3 times to obtain 3.2 g of white solid (i.e., compound H). The yield was 74%. 1H NMR (400 MHz, CDCl3) δ 8.15 (d, J=8.1 Hz, 2H), 8.05 (d, J=8.1 Hz, 2H), 7.67 (s, 1H), 5.41 (s, 2H), 3.96 (s, 3H); 13C NMR (100 MHz, CDCl3) δ192.06, 166.05, 161.55, 137.93, 134.56, 133.53, 130.03, 128.07, 53.71, 52.55.
4-(2-(2-amino-1,3, 4-thiadiazole) acetyl) methyl benzoate (3 g, 10.8 mmol) and 2N hydrochloric acid (60 mL) were added into 100 mL round-bottom flask for reflux reaction for 6 h at 100′C. TLC detection of raw material reaction is complete. The reaction bottle was cooled to room temperature, 1N sodium hydroxide was added under the ice bath to adjust the pH to 9-10, ethyl acetate was added, transferred to the separating funnel, Shake well and let it settle, the water layer was separated, the pH of the water layer was adjusted to 2-3 with 1N hydrochloric acid, and the solid (that is, compound I, 2.2 g) was extracted and filtered, the yield was 83%. 1H NMR (400 MHz, DMSO-d6) δ9.24 (s, 1H), 8.87 (s, 1H), 7.98-7.92 (m, 4H); 13C NMR (100 MHz, DMSO-d6) δ167.62, 151.69, 145.50, 145.26, 138.49, 130.38, 129.83, 125.12, 112.39.
compound C (50 mg, 0.12 mmol), 4-cyanophenylboronic acid (35.2 mg, 0.24 mmol), Pd(PPh3)4 (28 mg, 0.025 mmol), CsF (51 mg, 0.24 mmol) were added into 100 mL round-bottom flask. Dioxane (40 mmol) and water (10 mL), N2 protection, 100° C. reaction for 6 h. TLC test material reaction is complete, cooled to room temperature after filtration yellow solid 16 mg (i.e., compound L), yield 30%. 1H NMR (400 MHz, DMSO-d6) δ 8.14 (m, 9H), 3.91 (s, 3H).
The compound D (2 g, 6.2 mmol), EDCI (2.4 g, 12.4 mmol), NHS (1.4 g, 12.4 mmol), morpholine (2.7 g, 31 mmol) were dissolved in DMF (50 mL). The reaction was carried out at room temperature until the reaction of the raw material was completely detected by TLC. After the DMF was removed by spin evaporation, the white solid was purified by column chromatography (CH2Cl2:CH3OH=50:1), and 1.8 g (i.e., intermediate 1) was obtained. The yield was 75%. 1H NMR (400 MHz, CDCl3) δ 7.97 (d, J=8.3 Hz, 2H), 7.58 (d, J=8.3 Hz, 2H), 7.39 (s, 1H), 3.61 (s, 4H), 3.51 (s, 2H), 3.30 (s, 2H). 13C NMR (101 MHz, CDCl3) 5168.95, 161.56, 144.10, 138.68, 133.74, 131.25, 128.14, 127.24, 96.26, 66.82, 48.17, 42.65.
Compound D (2 g, 6.2 mmol), EDCI (2.4 g, 12.4 mmol), NHS (1.4 g, 12.4 mmol), 4-dimethylaminopiperidine (1.6 g, 12.4 mmol) were dissolved in DMF (50 mL), The reaction was carried out at room temperature until the reaction of the raw material detected by TLC was complete. After removal of DMF by rotary evaporation, it was purified by column chromatography (CH2C12:CH3OH=50:1), and 1.6 g of white solid (intermediate 2) was obtained with a yield of 59.3%. 1H NMR (400 MHz, DMSO-d6) δ 8.01 (d, J=8.4 Hz, 2H), 7.60 (d, J=8.0 Hz, 2H), 7.46 (s, 1H), 4.45 (d, J=13.1 Hz, 1H), 3.56 (d, J=13.6 Hz, 1H), 3.06 (t, J=12.9 Hz, 1H), 2.83 (t, J=12.6 Hz, 1H), 2.47-2.37 (m, 1H), 2.21 (s, 6H), 1.86 (d, J=12.9 Hz, 1H), 1.70 (d, J=12.6 Hz, 1H), 1.39 (q, J=13.5 Hz, 2H); 13C NMR (101 MHz, DMSO-d6) δ168.13, 162.63, 144.34, 140.26, 133.98, 130.48, 128.44, 127.58, 96.51, 61.73, 46.71, 41.78, 28.74, 28.10.
Compound D 8 mg (0.18 mmol), (1-ethyl-3 (3-dimethylpropylamine) carbodiimide 69 mg (0.36 mmol), N-hydroxysuccinimide 42 mg (0.36 mmol) were mixed in 10 mL DMF. After the reaction was stirred for 10 h at room temperature, 99 μL N-methylpiperazine was added, and after stirring for 1 h, the mixture was washed with water, extracted with chloroform, and column chromatography was performed to obtain 50 mg of yellow solid (intermediate 3), with a yield of 68%. 1H NMR (400 MHz, CDCl3) δ7.97-7.93 (m, 1H), 7.57-7.52 (m, 2H), 7.25 (s, 1H), 3.84-3.77 (m, 2H), 3.52-3.34 (m, 2H) 2H), 2.57-2.45 (m, 2H), 2.35 (s, 2H), 2.31 (s, 3H). 13C NMR (100 MHz, CDCl3) δ168.8, 161.7, 144.1, 139.2, 133.7, 131.0, 128.1, 127.2, 96.2, 55.2, 54.6, 47.6, 46.0, 42.1.
Compound I (2 g, 8.2 mmol), EDCI (3.2 g, 16.4 mmol), NHS (1.9 g, 16.4 mmol), morpholine (3.6 g, 41 mmol) were dissolved in DMF (50 mL), The reaction was carried out at room temperature until the reaction of the raw material detected by TLC was complete. The DMF was removed by rotary evaporation and purified by column chromatography (CH2Cl2:CH3OH=50:1) to obtain 1.8 g of white solid (intermediate 4) with a yield of 69.8%. 1H NMR (400 MHz, CDCl3)(8.57 (s, 1H), 8.14 (s, 1H), 7.88 (d, J=8.1 Hz, 2H), 7.47 (d, J=8.0 Hz, 2H), 3.72 (s, 6H), 3.56 (s, 2H); 13C NMR (100 MHz, CDCl3)(170.22, 147.03, 146.49, 144.67, 135.43, 134.24, 127.80, 125.18, 110.13, 66.89, 48.27, 42.68.
Compound I (2 g, 8.2 mmol) and N-methylpiperazine (1.6 g, 16.4 mmol) were used as raw materials to obtain a white solid (1.6 g) according to the synthesis method of intermediate 4, and the yield was 59.6%. 1H NMR (400 MHz, CDCl3)(8.56 (s, 1H), 8.14 (s, 1H), 7.87 (d, J=8.3 Hz, 2H), 7.47 (d, J=8.3 Hz, 2H), 3.82 (s, 2H), 3.51 (s, 2H), 2.50 (s, 2H), 2.39 (s, 2H), 2.34 (s, 3H); 13C NMR (100 MHz, CDCl3)(170.13, 146.80, 146.70, 144.61, 135.22, 134.82, 127.76, 125.15, 110.08, 55.18, 47.73, 46.04, 42.15.
Compound I (2 g, 8.2 mmol) and 4-dimethylaminopiperidine (2.1 g, 16.4 mmol) were used as raw materials to obtain a white solid (1.5 g) according to the synthesis method of intermediate 4 in a yield of 51.5%. 1H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 8.17 (s, 1H), 7.87 (d, J=8.2 Hz, 2H), 7.46 (d, J=8.2 Hz, 2H), 4.76 (s, 1H), 3.92 (s, 1H), 3.06 (s, 1H), 2.82 (s, 1H), 2.53 (ddt, J=11.4, 7.5, 3.6 Hz, 1H), 2.35 (s, 6H), 1.87 (s, 2H), 1.52 (s, 2H); 13C NMR (100 MHz, CDCl3)(170.52, 147.55, 146.49, 144.95, 135.26, 134.73, 127.62, 125.29, 110.34, 62.19, 47.17, 41.66, 41.27, 28.78, 27.63.
Compound I (2 g, 8.2 mmol) and 4-morpholine piperidine hydrochloride (3.4 g, 16.4 mmol) were used as raw materials to obtain a white solid (900 mg) according to the synthesis method of intermediate 4 in a yield of 27.6%. 1H NMR (400 MHz, CDCl3-CD3OD) δ8.59 (s, 1H), 8.10 (s, 1H), 7.80 (d, J=8.2 Hz, 2H), 7.40 (d, J=8.2 Hz, 2H), 4.67 (s, 1H), 3.85 (s, 1H), 3.71 (t, J=4.7 Hz, 4H), 3.01 (s, 1H), 2.75 (s, 1H), 2.59 (t, J=4.8 Hz, 4H), 2.52 (s, 1H), 1.97 (s, 1H), 1.83 (s, 1H), 1.46 (s, 2H); 13C NMR (100 MHz, CDCl3-CD3OD) δ172.74, 170.12, 147.22, 146.08, 144.60, 134.86, 134.34, 127.27, 124.95, 109.98, 66.36, 61.72, 49.24, 46.70, 41.26, 25.06.
Compound L (340 mg, 1.01 mmol) and LiOH·H2O (420 mg, 10 mmol) were mixed in 30 mL THF/H2O (V:V=1:1) and stirred for 12 h at room temperature. After adjusting the pH to 2-3 with HCl, the white solid compound M, intermediate 8, was filtered and used in the next step without further purification.
(4-(5-bromoimidazo[2,1-b][1,3,4]thiadiazol-2-yl)phenyl)(morpholino)methanone (intermediate 1) (200 mg, 0.5 mmol), 4-cyanophenylboronic acid (150 mg, 1 mmol), Reactions were performed with Pd(PPh3)4 (120 mg, 0.1 mmol), CsF (380 mg, 2.5 mmol), dioxane (40 mmol) and water (10 mL) under N2 protection at 100° C. for 6 h. Dichloromethane and water were added and transferred to a separating funnel. The organic layer was separated. The aqueous layer was extracted twice with dichloromethane, the organic layer was combined, the organic layer was washed twice with saturated NaCl solution, anhydrous Mg(SO4)2 was dried, filtered, and the filtrate was dried. After purification by column chromatography (CH2Cl2:CH3OH=50:1), 40 mg of white solid was obtained in 19.2% yield. 1H NMR (500 MHz, CDCl3)(8.14 (d, J=6.9 Hz, 2H), 8.02 (d, J=7.2 Hz, 2H), 7.77 (d, J=7.8 Hz, 3H), 7.62 (d, J=7.3 Hz, 2H), 3.82 (s, 4H), 3.68 (s, 2H), 3.48 (s, 2H); 13C NMR (125 MHz, CDCl3)(168.85, 162.00, 146.97, 138.84, 132.76, 132.52, 132.06, 131.13, 128.58, 128.25, 127.21, 124.83, 118.78, 110.66, 66.81, 48.18, 42.62; HRMS calcd for (C22H18O2N5S+H)+ 416.1176, found 416.1168.
(4-(5-bromoimidazo[2,1-b][1,3,4]thiadiazol-2-yl)phenyl)(morpholino)methanone (intermediate 1) (200 mg, 0.5 mmol), 4-fluorobenzoboronic acid (140 mg, 1 mmol) were added to a 100-ml solanaceous flask, Reactions with Pd(PPh3)4 (120 mg, 0.1 mmol), CsF (380 mmol, 2.5 mmol), dioxane (40 mmol) and water (10 mL) were performed under N2 protection at 100° C. for 6 h. Dichloromethane and water were added and transferred to a separating funnel. The organic layer was separated. The aqueous layer was extracted twice with dichloromethane, the organic layer was merged, the organic layer was washed twice with saturated NaCl solution, anhydrous Mg(SO4)2 was dried, filtered, and the filtrate was spun dry. After purification by column chromatography (CH2Cl2:CH3OH=50:1), 56 mg of white solid was obtained in a yield of 27.5%. 1H NMR (500 MHz, CDCl3) δ8.00 (d, J=6.4 Hz, 2H), 7.98-7.94 (m, 2H), 7.59 (d, J=6.4 Hz, 3H), 7.20 (t, J=7.8 Hz, 2H), 3.82 (s, 4H), 3.68 (s, 2H), 3.48 (s, 2H); 13C NMR (125 MHz, CDCl3) δ168.92, 163.29, 161.50, 144.92, 138.62, 135.70, 131.33, 129.69, 128.17, 127.16, 126.99, 124.36, 116.08, 114.45, 66.82, 48.12, 42.64; HRMS calcd for (C21H18O2N4FS+H)+409.1129, found 409.1119.
(4-(5-(4-hydroxyphenyl)imidazo[2,1-b][1,3,4]thiadiazol-2-yl)phenyl)(morpholino)methanone (intermediate 1) (200 mg, 0.5 mmol), 4-hydroxyphenylboronic acid (140 mg, 1 mmol), Reactions with Pd(PPh3)4 (120 mg, 0.1 mmol), CsF (380 mmol, 2.5 mmol), dioxane (40 mmol) and water (10 mL) were performed under N2 protection at 100° C. for 6 h. Dichloromethane and water were added and transferred to a separating funnel. The organic layer was separated. The aqueous layer was extracted twice with dichloromethane, the organic layer was merged, the organic layer was washed twice with saturated NaCl solution, anhydrous Mg(SO4)2 was dried, filtered, and the filtrate was spun dry. After purification by column chromatography (CH2Cl2:CH3OH=50:1), 60 mg of white solid was obtained in 29.5% yield. 1H NMR (500 MHz, DMSO-d6) δ 9.69 (s, 1H), 8.08 (d, J=7.9 Hz, 2H), 7.87 (d, J=8.2 Hz, 2H), 7.64 (d, J=8.2 Hz, 3H), 6.91 (d, J=8.8 Hz, 2H), 3.65 (s, 4H), 3.57 (s, 2H), 3.36 (s, 2H); 13C NMR (125 MHz, DMSO-d6) δ168.41, 161.20, 157.55, 139.21, 131.02, 129.93, 128.65, 128.05, 127.47, 126.71, 119.54, 116.22, 66.48, 48.08, 42.48; HRMS calcd for (C21H19O3N4S+H)+407.1172, found 407.1172.
Using intermediate 1 (200 mg, 0.5 mmol) and 4-methoxycarbonyl phenylboronic acid (180 mg, 1 mmol) as raw materials, 78 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 34.8%. 1H NMR (400 MHz, CDCl3)(8.15 (d, J=8.5 Hz, 3H), 8.08 (d, J=8.6 Hz, 3H), 8.02 (d, J=8.4 Hz, 2H), 7.77 (s, 1H), 7.60 (d, J=8.4 Hz, 2H), 3.95 (s, 3H), 3.82 (s, 4H), 3.66 (s, 2H), 3.48 (s, 2H); 13C NMR (100 MHz, CDCl3)(169.02, 166.77, 161.97, 146.13, 138.83, 133.72, 132.34, 131.25, 130.40, 129.11, 128.79, 128.35, 127.36, 124.63, 66.93, 52.37, 48.28, 42.75; HRMS calcd for (C23H21O4N4S+H)+ 499.1278, found 499.1269.
Using intermediate 1 (200 mg, 0.5 mmol) and 4-trifluoromethylphenylboronic acid (190 mg, 1 mmol) as raw materials, 80 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 34.9%. 1H NMR (400 MHz, CDCl3) δ 8.12 (d, J=8.1 Hz, 2H), 8.01 (d, J=8.1 Hz, 2H), 7.73 (d, J=8.1 Hz, 3H), 7.59 (d, J=8.2 Hz, 2H), 3.82 (s, 4H), 3.66 (s, 2H), 3.48 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 168.98, 161.50, 146.46, 138.67, 132.29, 131.80, 131.41, 129.44, 128.24, 127.20, 125.92, 124.85, 122.76, 66.85, 48.22, 42.77; HRMS calcd for (C22H18O2N4F3S+H)+459.1097, found 459.1086.
Using intermediate 1 (200 mg, 0.5 mmol) and 4-chlorobenylboronic acid (160 mg, 1 mmol) as raw materials, 76 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 35.8%. 1H NMR (400 MHz, CDCl3)(8.00 (d, J=8.6 Hz, 2H), 7.93 (d, J=8.7 Hz, 2H), 7.61 (s, 1H), 7.58 (d, J=8.6 Hz, 2H), 7.45 (d, J=8.7 Hz, 2H), 3.80 (s, 4H), 3.67 (s, 2H), 3.51 (s, 2H), 3.47 (s, 2H); 13C NMR (100 MHz, CDCl3)(169.02, 161.15, 145.70, 138.53, 133.34, 131.54, 131.23, 129.14, 128.20, 127.17, 126.94, 126.18, 66.85, 48.23, 42.58; HRMS calcd for (C21H18O2N4ClS+H)+425.0834, found 425.0823.
With intermediate 1 (200 mg, 0.5 mmol) and 4-methylphenylboronic acid (140 mg, 1 mmol) as raw materials, 64 mg of white solid was obtained according to the synthesis method of compound 1, and the yield was 31.7%. 1H NMR (400 MHz, CDCl3) δ 8.00 (d, J=8.6 Hz, 2H), 7.87 (d, J=8.2 Hz, 2H), 7.58 (m, 3H), 7.30 (d, J=7.9 Hz, 3H), 3.80 (s, 4H), 3.67 (s, 2H), 3.50 (s, 2H), 2.42 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 169.11, 160.63, 145.06, 138.33, 137.64, 131.79, 130.55, 129.59, 128.33, 128.13, 127.14, 125.63, 125.09, 66.86, 48.14, 42.67, 21.36; HRMS calcd for (C22H21O2N4S+H)+405.1380, found 405.1374.
54 mg of white solid was obtained from intermediate 1 (200 mg, 0.5 mmol) and 4-hydroxymethylphenylboronic acid (152 mg, 1 mmol) according to the synthesis method of compound 1, and the yield was 25.7%. 1H NMR (400 MHz, CDCl3) δ 7.95 (dd, J=8.4, 5.1 Hz, 4H), 7.58 (s, 1H), 7.54 (d, J=8.4 Hz, 2H), 7.46 (d, J=8.2 Hz, 2H), 4.74 (s, 2H), 3.81 (s, 4H), 3.65 (s, 2H), 3.46 (s, 2H), 2.40 (s, 1H); 13C NMR (100 MHz, CDCl3) δ 169.19, 161.00, 145.43, 140.62, 138.36, 131.65, 130.86, 128.21, 128.01, 127.58, 127.19, 125.19, 66.92, 65.02, 48.31, 42.72; HRMS calcd for (C22H21O3N4S+H)+421.1329, found 421.1322.
Using intermediate 1 (200 mg, 0.5 mmol) and 3-fluoro-4-trifluoromethylphenylboronic acid (208 mg, 1 mmol) as raw materials, 78 mg of white solid was obtained according to the synthesis method of compound 1, and the yield was 32.8%. 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J=8.2 Hz, 2H), 7.92 (dd, J=11.8, 1.6 Hz, 1H), 7.83 (d, J=8.3 Hz, 1H), 7.74 (s, 1H), 7.69 (t, J=7.8 Hz, 1H), 7.60 (d, J=8.2 Hz, 2H), 3.81 (s, 4H), 3.67 (s, 2H), 3.47 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 168.91, 161.95, 147.00, 138.85, 134.21, 133.03, 131.18, 128.29, 127.80, 127.24, 125.74, 119.84, 112.71, 66.84, 48.24, 42.67; HRMS calcd for (C22H17O2N4F4S+H)+ 477.1003, found 477.0989.
Using intermediate 1 (200 mg, 0.5 mmol) and 3-fluoro-4-hydroxyphenylboronic acid (156 mg, 1 mmol) as raw materials, 98 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 46.2%. 1H NMR (400 MHz, CDCl3-CD3OD) δ 7.94 (d, J=8.3 Hz, 2H), 7.66 (dd, J=12.2, 2.1 Hz, 1H), 7.51 (dd, J=8.6, 2.5 Hz, 3H), 7.41 (s, 1H), 6.98 (t, J=8.7 Hz, 1H), 3.75 (s, 4H), 3.60 (s, 2H), 3.42 (s, 2H); 13C NMR (100 MHz, CDCl3-CD3OD) δ 169.60, 161.23, 152.93, 150.54, 144.95, 138.31, 131.76, 129.74, 128.24, 127.41, 121.70, 120.44, 118.34, 113.07, 66.91, 42.84; HRMS calcd for (C21H18O3N4FS+H)+425.1078, found 425.1066.
Using intermediate 1 (200 mg, 0.5 mmol) and 3, 4-difluorobenboronic acid (160 mg, 1 mmol) as raw materials, 68 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 31.9%. 1H NMR (400 MHz, CDCl3) δ 8.00 (d, J=8.3 Hz, 2H), 7.87 (ddd, J=11.6, 7.5, 2.2 Hz, 1H), 7.68 (dddd, J=8.7, 3.9, 2.2, 1.4 Hz, 1H), 7.59 (m, 3H), 7.26 (dt, J=10.1, 8.5 Hz, 1H), 3.80 (s, 4H), 3.67 (s, 2H), 3.48 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 169.00, 161.50, 151.90, 150.99, 149.43, 148.50, 145.76, 138.63, 131.38, 128.23, 127.20, 126.29, 125.49, 121.11, 118.03, 114.14, 66.84, 48.21, 42.69; HRMS calcd for (C21H17O2N4F2S+H)+427.1035, found 427.1024.
From intermediate 2 (300 mg, 0.7 mmol) and 4-cyanophenylboronic acid (200 mg, 1.4 mmol), 50 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 15.6%. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J=8.1 Hz, 2H), 8.01 (d, J=7.9 Hz, 2H), 7.80-7.73 (m, 3H), 7.60 (d, J=7.9 Hz, 2H), 4.82 (s, 1H), 3.86 (s, 1H), 3.12 (s, 1H), 2.80 (m, 1H), 2.71 (s, 1H), 2.50 (s, 6H), 2.06 (s, 2H) 1.62 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 168.82, 161.88, 147.05, 139.18, 133.01, 132.78, 132.64, 131.13, 130.45, 128.06, 127.20, 126.58, 126.37, 124.79, 118.86, 110.53, 62.29, 46.59, 41.28, 40.89, 28.62, 26.98; HRMS calcd for (C25H25ON6S+H)+457.1805, found 457.1792.
From intermediate 2 (300 mg, 0.7 mmol) and 4-fluorobenboronic acid (200 mg, 1.4 mmol), 40 mg of white solid was obtained according to the synthesis method of compound 1, and the yield was 12.7%. 1H NMR (400 MHz, DMSO-d6) δ 8.09 (m, 4H), 7.82 (s, 1H), 7.61 (d, J=8.0 Hz, 2H), 7.36 (d, J=7.9 Hz, 2H), 4.46 (s, 1H), 3.56 (s, 1H), 3.06 (s, 1H), 2.83 (s, 1H), 2.26 (s, 6H), 1.89 (s, 1H), 1.74 (s, 1H), 1.41 (s, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.29, 161.80, 160.64, 145.44, 139.58, 131.69, 130.92, 128.45, 127.57, 127.18, 126.78, 125.23, 116.59, 116.37, 62.21, 55.06, 29.51, 26.30; HRMS calcd for (C24H25ON5FS+H)+450.1758, found 450.1750.
From intermediate 2 (300 mg, 0.7 mmol) and 4-hydroxyphenylboronic acid (190 mg, 1.4 mmol), 50 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 15.9%. 1H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1H), 8.04 (d, J=8.2 Hz, 2H), 7.84 (d, J=8.7 Hz, 2H), 7.63 (s, 1H), 7.59 (d, J=8.2 Hz, 2H), 6.88 (d, J=8.7 Hz, 2H), 4.46 (s, 1H), 3.53 (s, 1H), 3.03 (s, 1H), 2.82 (s, 1H), 2.51 (m, 1H), 2.24 (s, 6H), 1.84 (s, 1H), 1.69 (s, 1H), 1.38 (s, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.17, 161.26, 157.60, 144.25, 139.90, 130.85, 129.92, 128.34, 128.06, 127.45, 126.71, 119.54, 116.25, 61.73, 46.60, 41.54, 41.05, 28.54, 27.89; HRMS calcd for (C24H26O2N5S+H)+448.1802, found 448.1798.
From intermediate 2 (300 mg, 0.7 mmol) and 4-methoxyphenylboronic acid (210 mg, 1.4 mmol), 40 mg light yellow solid was obtained according to the synthesis method of compound 1, and the yield was 12.4%. 1H NMR (400 MHz, CDCl3) δ 7.98 (d, J=8.4 Hz, 2H), 7.90 (d, J=8.9 Hz, 2H), 7.55 (d, J=8.5 Hz, 2H), 7.51 (s, 1H), 7.02 (d, J=8.9 Hz, 2H), 4.78 (s, 1H), 3.87 (s, 4H), 2.85 (s, 1H), 2.70 (m, 1H), 2.44 (s, 6H), 2.00 (s, 2H), 1.54 (s, 2H); 13C NMR (101 MHz, CDCl3) δ 169.05, 160.73, 159.24, 144.76, 138.81, 132.58, 131.70, 129.93, 127.98, 127.14, 126.65, 121.23, 114.40, 62.37, 55.47, 41.11, 29.80; HRMS calcd for (C25H28O2N5S+H)+462.1958, found 462.1947.
From intermediate 2 (300 mg, 0.7 mmol) and 4-trifluoromethylphenylboronic acid (260 mg, 1.4 mmol), 32 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 9.2%. 1H NMR (400 MHz, CDCl3) δ 8.11 (d, J=8.1 Hz, 2H), 7.99 (d, J=8.0 Hz, 2H), 7.72 (d, J=9.7 Hz, 3H), 7.57 (d, J=8.6 Hz, 2H), 4.76 (s, 1H), 3.80 (s, 1H), 3.07 (s, 2H), 2.58 (m, 1H), 2.39 (s, 6H), 2.00 (s, 2H), 1.56 (m, 2H); 13C NMR (100 MHz, CDCl3) δ 168.87, 161.55, 146.44, 139.10, 132.24, 131.79, 131.28, 128.01, 127.15, 126.83, 125.94, 124.84, 62.31, 46.67, 41.04, 31.93, 29.71, 29.37; HRMS calcd for (C25H25ON5F3S+H)+500.1726, found 500.1718.
45 mg of white solid was obtained from intermediate 3 (200 mg, 0.5 mmol) and 4-cyanophenylboronic acid (150 mg, 1 mmol) according to the synthesis method of compound 1, and the yield was 21%. 1H NMR (400 MHz, CDCl3) δ 8.13 (d, J=8.3 Hz, 2H), 7.99 (d, J=7.9 Hz, 2H), 7.75 (d, J=8.3 Hz, 3H), 7.59 (d, J=8.0 Hz, 2H), 3.83 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.36 (s, 2H), 2.33 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.78, 161.94, 147.06, 139.34, 133.01, 132.77, 132.65, 131.01, 128.20, 127.15, 126.37, 124.79, 118.85, 110.55, 55.23, 54.64, 47.65, 46.02, 42.18; HRMS calcd for (C23H21ON6S+H)+429.1492, found 429.1485.
With intermediate 3 (200 mg, 0.5 mmol) and 4-fluorobenboronic acid (140 mg, 1 mmol) as raw materials, 60 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 28.6%. 1H NMR (400 MHz, CDCl3) δ 7.97 (m, 4H), 7.56 (m, 3H), 7.17 (m, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.52 (s, 2H), 2.37 (s, 2H), 2.37 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.99, 163.51, 161.27, 161.05, 145.36, 139.06, 131.43, 130.65, 128.20, 127.42, 127.17, 126.94, 124.77, 116.17, 115.95, 55.32, 54.71, 47.70, 46.12, 42.18; HRMS calcd for (C22H21ON5FS+H)+422.1445, found 422.1437.
46 mg of white solid was obtained from intermediate 3 (200 mg, 0.5 mmol) and 4-methoxycarbonyl phenylboronic acid (180 mg, 1 mmol) according to the synthesis method of compound 1 in a yield of 20%. 1H NMR (400 MHz, CDCl3) δ 8.14 (d, J=8.6 Hz, 2H), 8.08 (d, J=8.7 Hz, 2H), 8.00 (d, J=8.3 Hz, 2H), 7.73 (s, 1H), 7.58 (d, J=8.3 Hz, 2H), 3.94 (s, 3H), 3.83 (s, 2H), 3.46 (s, 2H), 2.52 (s, 2H), 2.38 (s, 2H), 2.34 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.95, 166.83, 161.54, 146.61, 139.20, 132.73, 132.56, 131.30, 130.37, 128.81, 128.24, 127.27, 127.22, 124.42, 55.33, 52.33, 47.73, 46.15, 42.25, 29.80; HRMS calcd for (C24H24O3N5S+H)+462.1594, found 462.1582.
From intermediate 3 (200 mg, 0.5 mmol) and pyridine-4-boronic acid (120 mg, 1 mmol), 50 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 24.7%. 1H NMR (400 MHz, CDCl3) δ 8.67 (d, J=6.3 Hz, 2H), 7.99 (d, J=8.4 Hz, 2H), 7.89 (d, J=6.3 Hz, 2H), 7.81 (s, 1H), 7.57 (d, J=8.5 Hz, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.36 (s, 2H), 2.32 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.86, 162.03, 150.51, 147.41, 139.33, 135.55, 133.49, 131.07, 128.27, 127.23, 125.53, 118.44, 55.30, 54.68, 47.69, 46.11, 42.20; HRMS calcd for (C21H20ON6S+H)+405.1458, found 405.1465.
54 mg of white solid was obtained from intermediate 3 (200 mg, 0.5 mmol) and 4-trifluoromethylphenylboronic acid (190 mg, 1 mmol) according to the synthesis method of compound 1, and the yield was 23%. 1H NMR (400 MHz, CDCl3) δ 8.10 (d, J=8.1 Hz, 2H), 7.98 (d, J=8.4 Hz, 2H), 7.71 (d, J=8.7 Hz, 3H), 7.57 (d, J=8.4 Hz, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.36 (s, 2H), 2.33 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.93, 161.71, 146.51, 139.25, 132.25, 131.83, 131.23, 128.25, 127.21, 126.88, 125.99, 124.91, 55.34, 54.72, 47.74, 46.15, 42.24; HRMS calcd for (C23H21ON5F3S+H)+472.1413, found 472.1407.
Using intermediate 3 (200 mg, 0.5 mmol) and 4-chlorobenylboronic acid (160 mg, 1 mmol) as raw materials, 56 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 25.6%. 1H NMR (400 MHz, CDCl3) δ 7.98 (d, J=8.3 Hz, 2H), 7.93 (d, J=8.6 Hz, 2H), 7.61 (s, 1H), 7.57 (d, J=8.3 Hz, 2H), 7.45 (d, J=8.6 Hz, 2H), 3.83 (s, 2H), 3.46 (s, 2H), 2.52 (s, 2H), 2.38 (s, 2H), 2.34 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.99, 161.37, 145.83, 139.18, 133.42, 131.43, 131.30, 129.24, 128.23, 127.26, 127.20, 127.07, 126.28, 55.38, 54.78, 47.78, 46.17, 42.30; HRMS calcd for (C22H21ON5ClS+H)+438.1150, found 438.1145.
From intermediate 3 (200 mg, 0.5 mmol) and 3-fluoro-4-trifluoromethylphenylboronic acid (210 mg, 1 mmol), 78 mg of white solid was obtained according to the synthesis method of compound 1, and the yield was 31.8%. 1H NMR (600 MHz, CDCl3) δ 8.00-7.89 (m, 3H), 7.83 (d, J=8.2 Hz, 1H), 7.73 (d, J=2.2 Hz, 1H), 7.68 (t, J=7.9 Hz, 1H), 7.57 (m, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.37 (m, 2H), 2.33 (m, 3H); 13C NMR (150 MHz, CDCl3) δ 168.86, 162.14, 147.09, 139.44, 134.28, 133.77, 133.07, 131.06, 128.29, 128.16, 127.25, 119.95, 112.59, 55.31, 54.72, 47.70, 46.09, 42.22; HRMS calcd for (C23H20ON5F4S+H)+ 490.1319, found 490.1307.
Using intermediate 3 (200 mg, 0.5 mmol) and 3-fluoro-4-cyanophenylboronic acid (160 mg, 1 mmol) as raw materials, 66 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 29.6%. 1H NMR (600 MHz, CDCl3) δ 7.99 (d, J=8.3 Hz, 2H), 7.96 (dd, J=10.4, 1.6 Hz, 1H), 7.85 (dd, J=8.1, 1.6 Hz, 1H), 7.78 (s, 1H), 7.69 (dd, J=8.2, 6.7 Hz, 1H), 7.59 (d, J=8.3 Hz, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.37 (s, 2H), 2.33 (s, 3H); 13C NMR (150 MHz, CDCl3) δ 168.79, 162.50, 147.71, 139.60, 135.22, 134.00, 130.87, 128.34, 127.28, 120.44, 114.11, 111.73, 55.33, 54.71, 47.70, 46.10, 42.22; HRMS calcd for (C23H20ON6FS+H)+ 477.1398, found 477.1400.
From intermediate 3 (200 mg, 0.5 mmol) and 4-nitrophenylboronic acid (170 mg, 1 mmol), 88 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 39.3%. 1H NMR (400 MHz, CDCl3) δ 8.35 (d, J=8.9 Hz, 2H), 8.20 (d, J=8.6 Hz, 2H), 8.02 (d, J=8.0 Hz, 2H), 7.83 (s, 1H), 7.61 (d, J=8.0 Hz, 2H), 3.84 (s, 2H), 3.48 (s, 2H), 2.53 (s, 2H), 2.39 (s, 2H), 2.35 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 167.43, 160.80, 146.13, 145.11, 138.12, 133.23, 132.28, 129.64, 126.91, 125.86, 124.84, 123.44, 123.17, 53.92, 53.37, 46.36, 44.72, 40.83; HRMS calcd for (C22H21O3N6S+H)+ 449.1390, found 449.1386.
Using intermediate 3 (200 mg, 0.5 mmol) and 3, 4-difluoro-phenylboronic acid (160 mg, 1 mmol) as raw materials, 78 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 35.6%. 1H NMR (400 MHz, CDCl3) δ 8.39-7.90 (m, 3H), 7.75-7.50 (m, 3H), 7.25 (s, 1H) 7.09-6.92 (m, 1H), 3.81 (s, 2H), 3.44 (s, 2H), 2.50 (s, 2H), 2.36 (s, 3H), 2.32 (d, J=1.0 Hz, 5H); 13C NMR (100 MHz, CDCl3) δ 168.86, 161.68, 161.11, 144.13, 139.23, 139.09, 134.16, 133.71, 131.31, 131.05, 128.09, 127.17, 111.78, 104.67, 55.23, 54.67, 47.64, 46.03, 42.18; HRMS calcd for (C22H20ON5F2S+H)+440.1351, found 440.1348.
A white solid 76 mg was obtained from intermediate 3 (200 mg, 0.5 mmol) and 2-cyano-5-pyridine boronic acid (150 mg, 1 mmol) according to the synthesis method of compound 1 in a yield of 35%. 1H NMR (400 MHz, CDCl3) δ 9.40 (d, J=2.3 Hz, 1H), 8.46 (ddd, J=8.2, 2.3, 0.8 Hz, 1H), 7.99 (d, J=8.5 Hz, 2H), 7.82-7.76 (m, 2H), 7.59 (d, J=8.6 Hz, 2H), 3.83 (s, 2H), 3.46 (s, 2H), 2.52 (s, 2H), 2.38 (s, 2H), 2.34 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.70, 162.68, 147.88, 146.78, 139.59, 133.72, 131.37, 130.72, 128.62, 128.27, 127.80, 127.20, 123.53, 117.34, 55.20, 54.60, 47.63, 46.01, 42.15; HRMS calcd for (C22H20ON7S+H)+430.1445, found 430.1435.
From intermediate 3 (200 mg, 0.5 mmol) and 2-fluoro-5-pyridine boronic acid (140 mg, 1 mmol), 65 mg white solid was obtained according to the synthesis method of compound 1, and the yield was 30.8%. 1H NMR (400 MHz, CDCl3) δ 8.88 (d, J=2.5 Hz, 1H), 8.36 (ddd, J=8.6, 7.5, 2.5 Hz, 1H), 7.98 (d, J=8.5 Hz, 2H), 7.65 (s, 1H), 7.58 (d, J=8.5 Hz, 2H), 7.07 (ddd, J=8.6, 3.1, 0.7 Hz, 1H), 3.83 (s, 2H), 3.46 (s, 2H), 2.50 (d, J=5.9 Hz, 2H), 2.38 (s, 2H), 2.34 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.83, 163.91, 161.87, 161.52, 146.18, 144.13, 139.25, 137.46, 131.39, 131.09, 128.18, 127.12, 124.19, 122.98, 110.08, 109.71, 55.21, 54.65, 47.60, 46.00, 42.13; HRMS calcd for (C21H20ON6FS+H)+ 423.1398, found 423.1393.
75 mg white solid was obtained from intermediate 3 (200 mg, 0.5 mmol) and 2-(trifluoromethyl) pyridyl-5-boronic acid (190 mg, 1 mmol) according to the synthesis method of compound 1, and the yield was 31.8%. 1H NMR (400 MHz, CDCl3) δ 9.35 (dd, J=2.3, 0.9 Hz, 1H), 8.45 (dd, J=8.2, 2.3 Hz, 1H), 7.98 (d, J=8.3 Hz, 2H), 7.83 (s, 1H), 7.78 (dd, J=8.2, 0.8 Hz, 1H), 7.58 (d, J=8.4 Hz, 2H), 3.82 (s, 2H), 3.45 (s, 2H), 2.51 (s, 2H), 2.38 (s, 2H), 2.33 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.75, 162.34, 147.31, 145.88, 139.48, 132.99, 132.37, 130.88, 128.22, 127.46, 127.14, 123.86, 120.66, 55.26, 54.61, 47.64, 46.03, 42.16; HRMS calcd for (C22H20ON6F3S+H)+473.1366, found 473.1355.
A mixture of intermediate 8 (65 mg, 0.18 mmol), EDCI (69 mg, 0.36 mmol), and NHS (42 mg, 0.36 mmol) with 10 mL DMF was stirred for 10 h at room temperature. Then pyrrolidine (65 μL, 0.9 mmol) was added and the reaction was continued for another 1 h. The reaction of the raw material was completely detected by TLC, quenched with water, extracted three times with DCM, the organic layer was washed three times with saturated NaCl solution, dried with anhydrous Mg(SO4)2, filtered, and the filtrate spun dry. Column chromatography purification (DCM:MeOH=15:1) was performed to give a white solid (50 mg) in 68% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.24 (d, J=8.3 Hz, 2H), 8.08-8.02 (m, 3H), 7.91 (d, J=8.3 Hz, 2H), 7.71 (d, J=8.1 Hz, 2H), 3.46 (t, J=6.6 Hz, 2H), 3.37 (t, J=6.2 Hz, 2H), 1.89-1.76 (m, 4H). 13C NMR (100 MHz, DMSO-d6) δ 167.5, 162.5, 147.3, 141.0, 134.3, 133.5, 132.8, 130.8, 128.7, 127.4, 126.1, 124.9, 119.4, 109.7, 49.3, 46.6, 26.5, 24.5. HRMS calculated for (M+H)+ 400.1227, found 400.1217.
From 1-(2-hydroxyethyl) pyrrolidine (104 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a white solid was obtained according to the synthesis method of compound 30 in a yield of 60%. 1H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J=8.2 Hz, 2H), 8.16-8.09 (m, 3H), 8.05 (d, J=8.2 Hz, 2H), 7.96 (d, J=8.2 Hz, 2H), 4.42 (dd, J=12.5, 6.3 Hz, 2H), 2.80 (t, J=6.8 Hz, 2H), 2.51-2.47 (m, 4H), 1.64-1.74 (m, 4H). HRMS calculated for (M+H)+ 443.1449, found 443.1452
From n-pentanthiol (91.8 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained according to the synthesis method of compound 30 in a yield of 53%. 1H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J=8.4 Hz, 2H), 8.04 (d, J=9.7 Hz, 3H), 8.01 (d, J=8.4 Hz, 2H), 7.96 (d, J=8.4 Hz, 2H), 3.53 (t, J=5.1 Hz, 1H), 1.97 (dd, J=11.5, 6.1 Hz, 2H), 1.82-1.77 (m, 4H), 1.32 (p, J=6.5 Hz, 2H), 0.8 (p, J=6.5 Hz, 2H). HRMS calculated for (M+H)+432.1176, found 432.1163.
A yellow solid was obtained from N-(3-hydroxypropyl) morpholine (131 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol) according to the synthesis method of compound 30 in a yield of 45%. 1H NMR (400 MHz, DMSO-d6) δ 8.17 (d, J=8.4 Hz, 2H), 8.03-8.09 (m, 3H), 7.82 (d, J=8.4 Hz, 2H), 7.71 (d, J=8.4 Hz, 2H), 3.50 (t, J=4.3 Hz, 4H), 3.35-3.30 (m, 2H), 2.41-2.31 (m, 2H), 1.72 (p, J=7.0 Hz, 2H).HRMS calculated for (M+H)+473.1528, found 473.1542.
From 1, 5-pentanediol (94 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained as described for compound 30 in a yield of 45%. 1H NMR (400 MHz, DMSO-d6) δ 8.19 (d, J=8.5 Hz, 2H), 8.10 (d, J=9.5 Hz, 3H), 7.96 (d, J=8.4 Hz, 2H), 7.87 (d, J=8.5 Hz, 2H), 3.21 (t, J=5.1 Hz, 2H), 3.41 (dd, J=11.6, 6.3 Hz, 2H), 1.69 (dd, J=12.9, 6.7 Hz, 2H), 1.31-1.22 (m, 2H), 1.06 (m, 2H).HRMS calculated for (M+H)+432.1347, found 432.1356.
From 2-mercaptoethanol (70.2 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained according to the synthesis method of compound 30 in a yield of 35%. 1H NMR (400 MHz, DMSO-d6) δ8.18 (d, J=8.4 Hz, 2H), 8.10-8.01 (m, 3H), 7.90 (d, J=8.5 Hz, 2H), 7.81 (d, J=8.4 Hz, 2H), 4.80 (t, J=5.6 Hz, 1H), 3.66 (q, J=6.0 Hz, 2H), 3.28 (dd, J=11.7, 5.8 Hz, 2H). HRMS calculated for (M+H)+ 406.0622, found 406.0635.
From N, N-dimethyl-1, 2-ethylenediamine (80 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained according to the synthesis method of compound 30 in a yield of 35%. 1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.26 (d, J=7.8 Hz, 2H), 8.10 (d, J=10.0 Hz, 3H), 8.03 (d, J=7.5 Hz, 2H), 7.94 (d, J=7.8 Hz, 2H), 3.37 (t, J=4.0 Hz, 2H), 2.43 (t, J=8.0 Hz, 2H), 2.20 (s, 6H). 13C NMR (100 MHz, DMSO-d6) δ 162.5, 147.4, 138.1, 134.4, 133.5, 132.9, 132.1, 128.9, 127.5, 126.1, 125.1, 119.5, 109.8, 100.00, 58.6, 45.8, 38.1. HRMS calculated for (M+H)+417.192, found, 417.1488.
A yellow solid was obtained from N-(3-aminopropyl) morpholine (130 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol) according to the synthesis method of compound 30 in 47% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.72 (t, J=5.4 Hz, 1H), 8.27 (d, J=8.4 Hz, 2H), 8.08-8.14 (m, 3H), 8.04 (d, J=8.4 Hz, 2H), 7.95 (d, J=8.4 Hz, 2H), 3.58 (t, J=4.3 Hz, 4H), 3.35-3.30 (m, 2H), 2.41-2.31 (m, 6H), 1.72 (p, J=7.0 Hz, 2H). 13C NMR (100 MHz, DMSO-d6) δ 165.5, 162.4, 147.3, 138.1, 134.3, 133.4, 132.8, 131.9, 128.7, 127.4, 126.0, 124.9, 119.4, 109.7, 66.7, 56.5, 53.8, 38.4, 26.4. HRMS calculated for (M+H)+473.1730, found 473.1742.
From 1-(2-aminoethyl) pyrrolidine (102 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a white solid was obtained as described for compound 30 in a yield of 62%. 1H NMR (400 MHz, DMSO-d6) δ 8.69 (t, J=5.2 Hz, 1H), 8.29 (d, J=8.2 Hz, 2H), 8.16-8.09 (m, 3H), 8.05 (d, J=8.2 Hz, 2H), 7.96 (d, J=8.2 Hz, 2H), 3.42 (dd, J=12.5, 6.3 Hz, 2H), 2.60 (t, J=6.8 Hz, 2H), 2.51-2.47 (m, 4H), 1.64-1.74 (m, 4H). 13C NMR (100 MHz, DMSO-d6) δ 165.5, 162.4, 147.2, 138.0, 134.4, 133.5, 132.9, 132.0, 128.8, 127.4, 126.1, 125.0, 119.4, 109.7, 55.3, 54.2, 23.6. HRMS calculated for (M+H)+443.1649, found 473.1642.
From 3-aminopropanol (68 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained as described for compound 30 in a yield of: 28%. 1H NMR (400 MHz, DMSO-d6) δ 8.70 (t, J=5.4 Hz, 1H), 8.28 (d, J=8.4 Hz, 2H), 8.12 (d, J=9.7 Hz, 3H), 8.05 (d, J=8.4 Hz, 2H), 7.96 (d, J=8.4 Hz, 2H), 4.53 (t, J=5.1 Hz, 1H), 3.50 (dd, J=11.5, 6.1 Hz, 2H), 3.40-3.35 (m, 2H), 1.72 (p, J=6.5 Hz, 2H). 13C NMR (100 MHz, DMSO-d6) δ 165.5, 162.3, 147.2, 138.1, 134.3, 133.4, 132.8, 131.9, 128.7, 127.3, 126.0, 124.9, 119.4, 109.7, 59.1, 37.3, 32.8. HRMS calculated for (M+H)+404.1176, found 404.1173.
From ethanolamine (55 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained as described for compound 30 in a yield of 36%. 1H NMR (400 MHz, DMSO-d6) δ 8.71 (t, J=5.5 Hz, 1H), 8.28 (d, J=8.4 Hz, 2H), 8.15-8.10 (m, 3H), 8.07 (d, J=8.5 Hz, 2H), 7.96 (d, J=8.4 Hz, 2H), 4.80 (t, J=5.6 Hz, 1H), 3.56 (q, J=6.0 Hz, 2H), 3.38 (dd, J=11.7, 5.8 Hz, 2H). 13C NMR (100 MHz, DMSO-d6) δ 165.7, 162.4, 147.3, 138.0, 134.3, 133.5, 132.9, 131.9, 128.8, 127.3, 126.0, 124.9, 119.4, 109.7, 60.1, 42.8. HRMS calculated for (M+H)+390.1019, found 390.1018.
From 2-methylpiperidine (89 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained as described for compound 30 in a yield of: 46%. 1H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J=8.5 Hz, 2H), 8.09 (d, J=8.6 Hz, 3H), 7.96 (d, J=8.5 Hz, 2H), 7.60 (d, J=8.1 Hz, 2H), 5.03-3.59 (m, 2H), 2.99 (d, J=37.9 Hz, 1H), 1.75-1.33 (m, 6H), 1.22 (d, J=6.8 Hz, 3H). 13C NMR (100 MHz, DMSO-d6) δ 168.4, 162.5, 147.2, 140.9, 134.3, 133.4, 132.9, 130.3, 127.8, 127.7, 126.0, 124.9, 119.4, 109.7, 30.2, 25.9, 18.9. HRMS calculated for (M+H)+428.1540, found 428.1533.
From 5-amino-1-amyl alcohol (93 mg, 0.9 mmol) and intermediate 8 (65 mg, 0.18 mmol), a yellow solid was obtained according to the synthesis method of compound 30 in yield: 48%. 1H NMR (400 MHz, DMSO-d6) δ 8.70 (t, J=5.4 Hz, 1H), 8.29 (d, J=8.5 Hz, 2H), 8.13 (d, J=9.5 Hz, 3H), 8.06 (d, J=8.4 Hz, 2H), 7.97 (d, J=8.5 Hz, 2H), 4.39 (t, J=5.1 Hz, 1H), 3.41 (dd, J=11.6, 6.3 Hz, 2H), 3.29 (dd, J=12.9, 6.7 Hz, 2H), 1.61-1.52 (m, 2H), 1.46 (dd, J=13.9, 6.8 Hz, 2H), 1.40-1.31 (m, 2H). 13C NMR (100 MHz, DMSO-d6) δ 165.4, 162.4, 147.3, 138.2, 134.3, 133.5, 132.8, 131.9, 128.8, 127.4, 126.0, 125.0, 119.4, 109.7, 61.1, 32.7, 29.5, 23.6. HRMS calculated for (M+H)+ 432.1489, found 432.1479.
Intermediate 4 (500 mg, 1.6 mmol), 4-cyanophenylacetylene (240 mg, 1.9 mmol), potassium tert-butyl alcohol (540 mg, 4.8 mmol), and DMF (40 mL) were added to a 50 mL tomato bottle. The reaction was carried out for 6 h at room temperature under magnetic stirring. The reaction of the raw material was complete as determined by TLC. Dichloromethane and water were added to the reaction flask, transferred to a separating funnel, shaken to rest, and the organic layer was separated. The aqueous layer was extracted twice with dichloromethane, the organic layer was merged, the organic layer was washed twice with saturated NaCl solution, anhydrous Mg(SO4)2 was dried, filtered, the filtrate was dried, and purified by column chromatography (CH2Cl2:CH3OH=50:1). White solid (200 mg) was obtained in 30.2% yield. 1H NMR (400 MHz, CDCl3) δ 7.94 (s, 1H), 7.89 (m, 1H), 7.87 (m, 3H), 7.81 (m, 2H), 7.46 (d, J=8.3 Hz, 2H), 6.98 (s, 1H), 3.76 (s, 6H), 3.52 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.29, 150.45, 147.16, 135.21, 134.26, 133.86, 133.28, 130.78, 127.84, 127.29, 125.39, 117.98, 113.43, 111.64, 107.75, 66.91, 48.08, 42.57; HRMS calcd for (C23H19O2N6S+H)+415.1223, found 415.1226.
A white solid (170 mg) was obtained from intermediate 4 (500 mg, 1.6 mmol) and 4-fluorophenylacetylene (230 mg, 1.9 mmol) according to the synthesis method of compound 30 in 26% yield. 1H NMR (400 MHz, CDCl3) δ 7.88 (m, 3H), 7.65 (dd, J=8.8, 5.1 Hz, 2H), 7.45 (d, J=8.3 Hz, 2H), 7.24 (m, 2H), 6.77 (s, 1H), 3.75 (s, 6H), 3.51 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.36, 164.64, 162.14, 150.41, 146.96, 135.73, 133.99, 131.57, 129.04, 128.96, 127.78, 126.03, 126.00, 125.25, 116.76, 116.54, 108.72, 107.71, 66.93, 48.10, 42.46; HRMS calcd for (C22H19O2N3FS+H)+ 408.1177, found 408.1178.
A white solid (280 mg) was obtained from intermediate 4 (500 mg, 1.6 mmol) and 4-methoxyphenylacetylene (250 mg, 1.9 mmol) according to the synthesis method of compound 30, and the yield was 41.7%. 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.00 (d, J=8.2 Hz, 2H), 7.77 (d, J=8.4 Hz, 2H), 7.44 (d, J=8.2 Hz, 2H), 7.35 (d, J=1.2 Hz, 1H), 7.13 (d, J=8.5 Hz, 2H), 3.85 (s, 3H), 3.73-3.47 (m, 8H); 13C NMR (100 MHz, DMSO-d6) δ 169.53, 162.85, 161.52, 160.58, 149.93, 146.21, 135.86, 134.35, 132.17, 128.84, 128.07, 125.20, 122.03, 115.17, 110.04, 108.62, 67.19, 66.21, 55.87, 45.58; HRMS calcd for (C23H22O3N3S+H)+ 420.1376, found 420.1370.
A white solid (220 mg) was obtained from intermediate 4 (500 mg, 1.6 mmol) and 4-trifluoromethylphenylacetylene (320 mg, 1.9 mmol) according to the synthesis method of compound 30, and the yield was 30.1%. 1H NMR (400 MHz, CDCl3) δ 7.93 (s, 1H), 7.89 (d, J=8.3 Hz, 2H), 7.82 (m, 4H), 7.46 (d, J=8.3 Hz, 2H), 6.93 (s, 1H), 3.76 (s, 6H), 3.52 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.31, 150.45, 147.19, 135.53, 134.15, 133.22, 131.19, 127.81, 127.22, 126.53, 125.30, 110.52, 107.73, 66.93, 48.32, 42.60; HRMS calcd for (C23H19O2N3S+H)+458.1145, found 458.1144.
A white solid (265 mg) was obtained from intermediate 4 (500 mg, 1.6 mmol) and 4-chlorophenylacetylene (260 mg, 1.9 mmol) according to the synthesis method of compound 30 in 39.1% yield. 1H NMR (400 MHz, CDCl3) δ 7.88 (d, J=4.4 Hz, 2H), 7.85 (s, 1H), 7.59 (d, J=8.6 Hz, 2H), 7.51 (d, J=8.6 Hz, 2H), 7.44 (d, J=8.4 Hz, 2H), 6.80 (s, 1H), 3.73 (s, 6H), 3.51 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.32, 150.40, 146.95, 135.77, 135.66, 134.01, 131.44, 129.71, 128.17, 127.78, 125.24, 109.22, 107.80, 66.92, 48.30, 42.71; HRMS calcd for (C22H19O2N3ClS+H)+ 424.0881, found 424.0870.
A white solid (280 mg) was obtained from intermediate 5 (500 mg, 1.5 mmol) and 4-cyanophenyl acetylene (230 mg, 1.8 mmol) according to the synthesis method of compound 30, and the yield was 43.7%. 1H NMR (400 MHz, CDCl3) δ 7.92 (s, 1H), 7.88-7.84 (m, 4H), 7.80 (d, J=8.7 Hz, 2H), 7.45 (d, J=8.6 Hz, 2H), 6.97 (s, 1H), 3.80 (s, 2H), 3.49 (s, 2H), 2.48 (s, 2H), 2.36 (s, 2H), 2.32 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 170.15, 150.41, 147.48, 135.24, 134.71, 133.99, 133.26, 130.77, 127.77, 127.26, 125.26, 118.00, 113.37, 111.39, 107.63, 55.05, 47.56, 45.96, 41.99; HRMS calcd for (C24H22ON5S+H)+428.1540, found 428.1535.
The white solid (220 mg) was obtained from intermediate 5 (500 mg, 1.5 mmol) and 4-fluorophenylacetylene (220 mg, 1.8 mmol) according to the synthesis method of compound 30, and the yield was 34.9%. 1H NMR (400 MHz, CDCl3) δ 7.85 (d, J=8.0 Hz, 3H), 7.64 (dd, J=8.8, 5.1 Hz, 2H), 7.43 (d, J=8.5 Hz, 2H), 7.22 (d, J=8.8 Hz, 2H), 6.75 (s, 1H), 3.79 (s, 2H), 3.49 (s, 2H), 2.48 (s, 2H), 2.35 (s, 2H), 2.31 (s, 3H); 13CNMR (100 MHz, CDCl3) δ 170.24, 164.61, 162.11, 150.35, 147.03, 135.51, 134.48, 131.55, 129.02, 127.71, 126.04, 125.17, 116.51, 108.65, 107.65, 55.21, 54.82, 47.70, 46.02, 42.15; HRMS calcd for (C23H22ON4FS+H)+ 421.1493, found 421.1490.
A white solid (300 mg) was obtained from intermediate 5 (500 mg, 1.5 mmol) and 4-trifluoromethylphenylacetylene (310 mg, 1.8 mmol) according to the synthesis method of compound 30 in 42.5% yield. 1H NMR (400 MHz, CDCl3) δ 7.92 (s, 1H), 7.86 (d, J=8.6 Hz, 2H), 7.80 (m, 4H), 7.44 (d, J=8.6 Hz, 2H), 6.91 (s, 1H), 3.79 (s, 2H), 3.49 (s, 2H), 2.47 (s, 2H), 2.36 (s, 2H), 2.32 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 168.66, 148.87, 145.72, 133.82, 133.04, 131.68, 129.63, 126.19, 125.66, 124.97, 123.68, 108.92, 106.15, 53.55, 46.07, 44.43, 40.52; HRMS calcd for (C24H22ON4F3S+H)+ 471.1461, found 471.1452.
The white solid (264 mg) was obtained from intermediate 5 (500 mg, 1.5 mmol) and 4-chlorophenylacetylene (245 mg, 1.8 mmol) according to the synthesis method of compound 30, and the yield was 40.3%. 1H NMR (400 MHz, CDCl3) δ 7.87 (s, 1H), 7.84 (d, J=8.5 Hz, 2H), 7.58 (d, J=8.7 Hz, 2H), 7.50 (d, J=8.7 Hz, 2H), 7.42 (d, J=8.5 Hz, 2H), 6.78 (s, 1H), 3.78 (s, 2H), 3.48 (s, 2H), 2.46 (s, 2H), 2.35 (s, 2H), 2.30 (s, 3H); 13C NMR (101 MHz, CDCl3) δ 170.22, 150.37, 147.04, 135.77, 135.45, 134.47, 131.44, 129.70, 128.23, 128.17, 127.70, 125.17, 109.15, 107.72, 55.18, 54.86, 47.68, 46.00, 42.09; HRMS calcd for (C23H22ON4ClS+H)+ 437.1197, found 437.1197.
The white solid (280 mg) was obtained from intermediate 6 (500 mg, 1.3 mmol) and 4-fluorophenylacetylene (190 mg, 1.6 mmol) according to the synthesis method of compound 30, and the yield was 43.9%. 1H NMR (400 MHz, CDCl3) δ 7.85 (m, 3H), 7.64 (dd, J=8.8, 5.1 Hz, 2H), 7.42 (d, J=8.6 Hz, 2H), 7.23 (d, J=8.7 Hz, 2H), 6.75 (s, 1H), 4.71 (s, 1H), 3.88 (s, 1H), 3.71 (t, J=4.6 Hz, 4H), 3.02 (s, 1H), 2.81 (s, 1H), 2.55 (t, J=4.6 Hz, 4H), 2.43 (m, 1H), 1.96 (s, 1H), 1.82 (s, 1H), 1.58 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.19, 164.62, 162.13, 150.36, 147.07, 135.44, 134.75, 131.56, 129.03, 128.95, 127.53, 126.02, 125.18, 116.75, 116.52, 108.64, 107.62, 67.14, 61.98, 49.81, 46.93, 41.55, 28.96, 28.15; HRMS calcd for (C27H2802N4FS+H)+ 491.1912, found 491.1908.
A white solid (215 mg) was obtained from intermediate 7 (500 mg, 1.4 mmol) and 4-fluorophenylacetylene (200 mg, 1.7 mmol) according to the synthesis method of compound 30, and the yield was 34.2%. 1H NMR (400 MHz, CDCl3) δ 7.85 (d, J=9.1 Hz, 3H), 7.64 (dd, J=8.8, 5.1 Hz, 2H), 7.42 (d, J=8.4 Hz, 2H), 7.22 (d, J=8.6 Hz, 2H), 6.75 (s, 1H), 4.73 (s, 1H), 3.87 (s, 1H), 3.01 (s, 1H), 2.80 (s, 1H), 2.48 (m, 1H), 2.34 (s, 6H), 2.03-1.78 (m, 2H), 1.48 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.22, 164.61, 162.12, 150.35, 147.05, 135.47, 134.69, 131.56, 129.02, 128.94, 127.54, 126.04, 126.01, 125.18, 116.73, 116.52, 108.64, 107.65, 62.26, 46.93, 41.38, 28.88, 27.86; HRMS calcd for (C25H26ON4FS+H)+ 499.1806, found 499.1795.
The white solid (284 mg) was obtained from intermediate 7 (500 mg, 1.4 mmol) and 4-trifluoromethylphenylacetylene (290 mg, 1.7 mmol) according to the synthesis method of compound 30, and the yield was 40.7%. 1H NMR (400 MHz, CDCl3) δ 7.91 (s, 1H), 7.85 (d, J=8.3 Hz, 2H), 7.79 (m, 4H), 7.42 (d, J=8.3 Hz, 2H), 6.91 (s, 1H), 4.73 (s, 1H), 3.87 (s, 1H), 3.01 (s, 1H), 2.80 (s, 1H), 2.49 (m, 1H), 2.34 (s, 6H), 1.97 (s, 2H), 1.49 (m, 2H); 13C NMR (100 MHz, CDCl3) δ 170.17, 150.40, 147.25, 135.31, 134.81, 133.22, 131.17, 127.56, 127.20, 126.50, 125.22, 122.33, 110.49, 107.72, 62.26, 46.96, 41.36, 28.88, 27.77; HRMS calcd for (C26H26ON4F3S+H)+ 499.1774, found 499.1783.
A white solid (254 mg) was obtained from intermediate 7 (500 mg, 1.4 mmol) and 4-chlorophenylacetylene (230 mg, 1.7 mmol) according to the synthesis method of compound 30 in a yield of 39%. 1H NMR (400 MHz, CDCl3) δ 7.87 (s, 1H), 7.84 (d, J=8.3 Hz, 2H), 7.59 (d, J=8.6 Hz, 2H), 7.50 (d, J=8.6 Hz, 2H), 7.41 (d, J=8.3 Hz, 2H), 6.79 (s, 1H), 4.73 (s, 1H), 3.88 (s, 1H), 3.01 (s, 1H), 2.79 (s, 1H), 2.52 (m, 1H), 2.35 (s, 6H), 1.88 (s, 2H), 1.49 (s, 2H); 13C NMR (100 MHz, CDCl3) δ 170.21, 150.38, 147.08, 135.79, 135.44, 134.66, 131.46, 129.72, 128.25, 128.18, 127.55, 125.19, 109.13, 107.71, 62.29, 46.89, 41.27, 28.70, 27.68; HRMS calcd for (C25H26ON4ClS+H)+ 465.1510, found 465.1501.
Thiocarbamide was mixed with methyl p-formyl benzoate and stirred in methanol for 2 hours at room temperature. Then FeCl3.6H2O dissolved in methanol was added, heated and stirred at 75° C. for 30 min, cooled to room temperature, and the solid was filtered and rinsed with methanol until the filtrate was colorless to obtain compound 0. Then the mixture with 2-bromo-1-(4-fluorophenyl) one, 2-bromo-1-(4-fluorophenyl) one or 2-bromo-1-(4-cyanophenyl) one was refluxed and stirred in acetonitrile for 12 h. The yellow intermediate was obtained by column chromatography, and the intermediate was refluxed and stirred in acetic acid for 5 h. The mixture was then poured into ice water and filtered to obtain the corresponding compounds P1 or P2 or P3. The mixtures of P1 or P2 or P3, respectively, and LiOH H2O were stirred in THF/H2O(1:1) at room temperature for 12 hours. After adjusting the pH to 2-3 with HCl, the yellow solid Q1 or Q2 or Q3 was filtered and used in the next step without further purification. compounds 51-56 were obtained by adding Q1, Q2, or Q3 with EDCI, NHS, and the corresponding amines to DMF solvent, mixing and stirring, and stirring for 12 h at room temperature.
Q2 (0.5 mmol), EDCI (1 mmol), NHS (1 mmol), and N,N,2, 2-tetramethyl-1, 3-propylene diamine (2.5 mmol) were added to a round-bottom flask, followed by solvent DMF, and the mixture was stirred for 12 h at room temperature. After the completion of the reaction as judged by TLC, the reaction mixture was diluted with water and extracted three times with ethyl acetate (20 mL). The bound organic layer was washed with water and brine, dried over anhydrous MgSO4, filtered, and concentrated in vacuo. The crude product was purified by chromatography (DCM:MeOH=10:1) to give a white solid compound 51 in 45% yield. 1H NMR (500 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.08 (s, 4H), 7.90 (d, J=7.2 Hz, 2H), 7.43 (t, J=7.7 Hz, 2H), 7.30 (t, J=7.4 Hz, 1H), 3.30 (s, 2H), 3.01 (s, 2H), 2.88 (s, 6H), 1.07 (s, 6H). 13C NMR (125 MHz, DMSO-d6) δ 166.9, 161.1, 159.1, 158.8, 158.5, 158.1, 145.9, 145.0, 137.3, 133.7, 132.4, 129.1 (d, J=3.4 Hz), 128.0, 127.0, 125.2, 118.8, 116.5, 114.2, 111.9, 111.2, 65.6, 47.2, 46.7, 36.3, 24.2. HRMS calculated for (M+H)+434.2009, found 434.2010.
From Q2 and 1-(3-aminopropyl) tetrahydropyrrole, a white solid compound 52 was obtained in 49% yield according to the synthesis method of compound 51. 1H NMR (500 MHz, DMSO-d6) δ 8.82-8.77 (m, 2H), 8.05 (q, J=8.4 Hz, 4H), 7.91 (d, J=7.6 Hz, 2H), 7.43 (t, J=7.7 Hz, 2H), 7.30 (t, J=7.3 Hz, 1H), 3.37-3.34 (m, 2H), 2.81 (d, J=86.5 Hz, 6H), 1.83 (s, 6H). 13C NMR (125 MHz, DMSO-d6) δ 165.8, 160.9, 146.3, 145.02, 137.5, 134.1, 132.3, 129.2, 128.8, 127.9, 127.1, 125.2, 111.1, 53.9, 52.9, 37.4, 26.9, 23.2. HRMS calculated for (M+H)+432.1853, found 432.1855.
From Q1 and 4-(2-aminoethyl) pyridine, a white solid compound 53 was obtained in 31% yield according to the synthesis method of compound 51. 1H NMR (500 MHz, DMSO-d6) δ 8.79 (t, J=5.6 Hz, 1H), 8.77 (s, 1H), 8.47 (d, J=5.9 Hz, 2H), 8.05 (d, J=8.4 Hz, 2H), 7.99 (d, J=8.4 Hz, 2H), 7.96-7.91 (m, 2H), 7.27 (dd, J=15.5, 6.7 Hz, 4H), 3.57 (dd, J=12.9, 6.9 Hz, 2H), 2.90 (dd, J=11.9, 4.7 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) δ 165.6, 162.1 (d, J=244.2 Hz), 160.9, 149.8, 148.9, 145.2 (d, J=29.8 Hz), 137.6, 132.2, 130.7 (d, J=2.8 Hz), 128.7, 127.2 (d, J=6.9 Hz), 124.7, 116.1 (d, J=21.6 Hz), 111.0, 109.9, 40.56, 34.57. HRMS calculated for (M+H)+444.1289, found 444.1291.
From Q2 and ethanolamine, a white solid compound 54 was obtained in a yield of 51% according to the synthesis method of compound 51. 1H NMR (500 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.66 (t, J=5.5 Hz, 1H), 8.05 (s, 4H), 7.91 (d, J=7.3 Hz, 2H), 7.43 (t, J=7.7 Hz, 2H), 7.30 (t, J=7.3 Hz, 1H), 4.76 (s, 1H), 3.53 (t, J=6.2 Hz, 2H), 3.36 (d, J=5.0 Hz, 2H). 13C NMR (125 MHz, DMSO-d6) δ 165.7, 160.9, 146.2, 145.0, 137.7, 134.1, 132.2, 129.2, 128.8, 127.9, 127.0, 125.2, 111.2, 60.1, 42.8. HRMS calculated for (M+H)+365.1067, found 365.1065.
The white solid compound 55 was obtained from Q3 and tetrahydropyrrole according to the synthesis method of compound 51, and the yield was 51%. 1H NMR (400 MHz, TFA-d1) δ 8.47 (s, 1H), 8.19 (d, J=7.9 Hz, 2H), 7.92 (d, J=8.8 Hz, 4H), 7.86 (d, J=7.9 Hz, 2H), 3.93 (s, 2H), 3.70 (s, 2H), 2.18 (d, J=6.3 Hz, 2H), 2.10 (d, J=5.9 Hz, 2H). 13C NMR (400 MHz, TFA-d1) 166.4, 145.3, 137.6, 133.5, 131.1, 130.2, 128.6, 128.1, 126.6, 51.8, 49.3, 24.8, 23.5.
The green solid compound 56 was obtained from Q3 and N-methylpiperazine in 63% yield according to the synthesis method of compound 51. 1H NMR (400 MHz, TFA-d1) δ 8.47 (s, 1H), 8.15 (d, J=8.1 Hz, 2H), 7.96-7.89 (m, 4H), 7.73 (d, J=8.1 Hz, 2H), 5.04 (d, J=14.5 Hz, 1H), 4.08 (d, J=14.0 Hz, 1H), 3.85 (d, J=6.8 Hz, 2H), 3.73 (d, J=12.0 Hz, 1H), 3.57 (t, J=13.0 Hz, 1H), 3.32 (dt, J=37.8, 11.3 Hz, 2H), 3.11 (s, 3H). 13C NMR (100 MHz, DMSO-d6) δ 168.2, 161.9, 145.8, 144.3, 139.7, 138.7, 133.3, 130.7, 128.6, 127.5, 125.7, 119.5, 113.4, 109.9, 55.0, 46.0.
MNK1 and MNK2 kinase inhibition assays were performed using LANCE Ultra kinase activity assay developed by PerkinElmer. Staurosporine (STSP) was used as a positive control in this assay. The test steps are as follows: At 25° C., 1.00 ng MNK1 or 0.05 ng MNK2 was prepared with different concentrations (10000, 1000, 100, 10, 1 nM) of the tested compound to form a total volume of 10 μL reaction mixture (MNK1: 12.5 nM CREB, 450 μM ATP, 2 mM DTT, 1× buffer; MNK2: 12.5 nM CREB, 100 μM ATP, 2 mM DTT, 1×buffer), and incubation for 60 min. The reaction was terminated by adding 5 μL EDTA/Detection buffer and 5 μL Eu-CREB/Detection buffer. The ratio of HTRF signals at 615 nm and 665 nm was detected after 60 min of incubation. The inhibition rate was calculated by measuring the compounds at 1000 nM. When the inhibition rate > At 75%, 50%, and 25%, they were recorded as +++, ++, and +, respectively, as shown in Table 1.
Then five different concentrations (10000, 1000, 100, 10, 1 nM) of the tested compounds were measured, and the IC50 value was calculated according to the inhibition rate fitting curve. In vitro test of compound of IC50 MNK1/2 results are shown in table 2, including IC50<0.05 uM (****), 0.05 to 0.2 uM (***), 0.2 to 0.8 uM (**), and 0.8 1.5 uM (*).
We tested the effects of the above compounds on different mouse disease models, illustrated below by compound 12 as a specific Example.
C57BL/6J male mice were fed with high-fat diet (60 kcal %) and low-fat diet (10 kcal %), respectively. After 16 weeks, the obesity model was successfully established (the body weight of high-fat diet was higher than 20% of the body weight of low-fat diet). They were divided into blank group, 12 high dose group (100 mg/kg), 12 low dose group (50 mg/kg) and orlistat group (50 mg/kg). The mice in the control group were given normal saline as the control, and the daily afternoon gavage of the drug for 4 weeks. During the period of regular measurement of food intake and body weight changes, the results showed that after 1 month of treatment, the body weight of the treated group decreased significantly, but the food intake did not decrease.
A total of 50 male Kunming mice were fed in a constant temperature of 25° C. with free access to food and water. After 5 days of adaptive feeding, the mice were divided into two groups for the first time. The first group (n=10) was the blank group (C) fed with normal diet, and the rest were the second group (n=40) fed with 60% high-fat diet to induce high-fat model and free access to water for 4 weeks. At the fifth week, the mice in the second group were injected with streptozotocin (STZ) to induce type 2 diabetes model. The total injection dose was 150 mg/kg, and the feeding conditions were unchanged. Type 2 diabetic mice with blood glucose levels higher than 11 mmol/L were divided into model group (M, n=10) and control group (M, n=10). Distilled water), metformin group (Meft, 225 mg/kg/d), compound 12 low-dose group (2A-L, 50 mg/kg/d), compound 12 high-dose group (2A-H, 100 mg/kg/d). The rats in group M were given distilled water by gavage every morning, and the dosage was Meft: 225 mg/kg. 2A-L: 50 mg/kg; 2A-H: 100 mg/kg. The mice were administrated by gavage at a dose of 0.2 mL/g for 8 weeks, and the body weight and fasting blood glucose were measured regularly during the period.
The results showed (
compound 12 improved glucose tolerance (
compound 12 significantly reduced serum AST levels (
Six-week-old male db/db mice were randomly divided into 6 groups of 12 mice in each group: Model group (M, distilled water), metformin group (Meft, 200 mg/kg/d), lovastatin group (6 mg/kg), compound 12 low dose group (20 mg/kg/d), compound 12 middle dose group (40 mg/kg/d), compound 12 high dose group (80 mg/kg/d). They were raised in a constant temperature of 25° C. and fed and drinking freely. The mice were administrated by gavage at a dose of 0.2 mL/g for 6 weeks, and the body weight and fasting blood glucose were measured regularly during the period.
Insulin tolerance test (ITT) was performed with normal saline (1.0 U/ml, 1.0 U/kg) and fasting for 4 hours in the morning. In the afternoon experiment, weight was weighed, blood glucose was measured before insulin injection, insulin injection volume was calculated according to body weight, and blood glucose was measured at 15 min, 30 min, 45 min, and 60 min. At the end of the experiment, each cage was supplemented with feed. As observed by insulin tolerance test (
In the serum biochemical indicators of liver function (
In the serum active protein index (
The above test results indicate that the compounds of the invention have good MNK1/2 protein kinase inhibitory effect and exhibit good safety in cells. Taking example compound 12 as a representative, this class of compounds can significantly control blood glucose, reduce body weight, improve various blood biochemical indicators, and protect important organs such as liver in diabetic mice and obese mice models, which has a good prospect for drug development and application.
The present invention is described in detail above, and specific examples are used in this article to describe the principles and embodiments of the present invention. The above examples are only used to help understand the methods of the present invention and its core ideas, including the best way, and also to enable any skilled person in the art to practice the present invention, including the manufacture and use of any device or system. And the implementation of any combined approach. It should be pointed out that for ordinary technicians in the art, the invention can also be improved and modified on the premise of not separated from the principle of the invention, and these improvements and modifications also fall within the scope of protection of the claims of the invention. The scope of protection of the invention patent is defined by the claims and may include other embodiments capable of being thought of by those skilled in the art. If these other embodiments have structural elements that are not different from the verbal representation of the claims, or if they include equivalent structural elements that are not materially different from the verbal representation of the claims, then these other embodiments shall also be included within the scope of the claims.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202111435801.7 | Nov 2021 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2022/133420 | 11/22/2022 | WO |
