Patent Application
20240299386
The present invention relates to a novel imidazole derivative having inhibitory activity against the protein kinase, c-Jun N-terminal kinase 3 (JNK3), and uses thereof.
The present invention was made under Project no. NRF-2019M3A9A8066500 under the support of the Ministry of Science and ICT (MSIT) of the Republic of Korea (2017Y); the research management agency for this project is “National Research Foundation of Korea,” the research project name is “Original Technology Development Project/Bio-medical Technology Development Project/New Drug Target Discovery and Verification Project,” the research project title is “Validation of INK inhibitor with effects of inhibiting nerve cell death and improving cognitive function as a therapeutic agent for Alzheimer's dementia”, and the research period is “2020.03.01.-2020.12.31.”
In addition, the present invention was made under Project no. NRF-2020R1A6A1A03042854 under the support by the Ministry of Education (2020Y); the research management agency for this project is “National Research Foundation of Korea,” the research project name is “Basic Research Project in the Field of Science and Engineering/Basic Research Infrastructure Construction Project/University Key Research Institute Support Project—Key Research Institute Support (Science and Engineering Fields),” the research project title is “Pharmaceutical technology research institute,” and the research period is “2021.03.01.-2022.02.28.”
This application claims priority to and the benefit of Korean Patent Application No. 10-2021-0108424, filed on Aug. 18, 2021, the disclosure of which is incorporated herein by reference in its entirety.
In the case of Alzheimer's dementia, which is manifested by fatal dysfunction of the nervous system and motor impairment, due to a lack of understanding of the pathological phenomenon, existing therapeutic agents are mainly acetylcholinesterase inhibitors that alleviate the worsening of symptoms. According to continuous research on pathological phenomena, recently, with the development of therapeutic agents for degenerative brain diseases, there has been a so-called paradigm shift that moves from symptom alleviation to the development of a novel target protein inhibitor, and at the center of this, there are protein kinases, which are intracellular signaling proteins that control the apoptosis and necrosis of nerve cells.
In a representative signaling system, the MAPK pathway, which controls cell apoptosis, unlike approximately 10 similar isozymes, it is known that the c-Jun N-terminal kinase 3 (JNK3) isoform is concentrated in brain tissue and thus is deeply related to the apoptosis of nerve cells, and is distributed in pyramidal neurons (yellow) located in the CA1 and CA2 parts of the cerebral cortex and hippocampus, causing apoptosis due to overexpression of c-Jun, which is one of the intraneuronal substances, the degree of phosphorylation by JNK3, and the hyperactivation of JNK3.
First, JNK3 phosphorylates and activates amyloid precursor protein (APP), which is a precursor of Aβ as the main cause of Alzheimer's dementia, to be located in the cell membrane and to promote conversion into Aβ42. Particularly, the endocytosis of phosphorylated APP in this process is known to be a critical step in Aβ42 production. Second, it is noteworthy that Tau, which is the main ingredient of NFT found in brain cells of Alzheimer patients, is also phosphorylated by JNK. In addition, it was reported that neurodegeneration induced by APP/Aβ is attenuated (neuroprotection) in JNK3 gene-deleted animal models (Tg2576/PSM146L transgenic mice), showing that JNK3 also mediates the production and toxicity of Aβ, and a JNK3 inhibitor can be effectively used as an AD therapeutic agent.
The present invention is provided to solve the above problems, in an effort to find novel materials having the potential to be developed as therapeutic agents for brain diseases such as Alzheimer's dementia, Parkinson's disease, and Huntington's disease, the present inventors tried to develop low-molecular inhibitors that target c-Jun N-terminal kinase (JNK3) to adjust its function, and thus identified a novel imidazole derivative exhibiting JNK3 inhibitory activity. Based on this, the present invention was completed.
The present invention is directed to providing a novel imidazole derivative exhibiting JNK3 inhibitory activity or a pharmaceutically acceptable salt thereof.
The present invention is also directed to providing a method of preparing a novel imidazole derivative exhibiting JNK3 inhibitory activity.
The present invention is also directed to providing a pharmaceutical composition for use in preventing or treating a degenerative brain disease, which includes the imidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.
The present invention is also directed to providing a method of treating a degenerative brain disease, which includes administering the imidazole derivative or a pharmaceutically acceptable salt thereof to an individual or subject in need thereof.
The present invention is also directed to providing an imidazole derivative or a pharmaceutically acceptable salt thereof, which is used to treat a degenerative brain disease.
The present invention is also directed to providing uses of the imidazole derivative or a pharmaceutically acceptable salt thereof to be used in the preparation of a drug for treating a degenerative brain disease.
However, technical problems to be solved in the present invention are not limited to the above-described problems, and other problems which are not described herein will be fully understood by those of ordinary skill in the art from the following descriptions.
To achieve the purpose of the present invention, a compound of Formula 1 below or a pharmaceutically acceptable salt thereof is provided.
In Formula 1,
In addition, the present invention provides a pharmaceutical composition for use in preventing or treating a degenerative brain disease, which includes the compound of Formula 1 above or a pharmaceutically acceptable salt thereof as an active ingredient.
In one embodiment of the present invention, the composition may inhibit the activity of JNK3.
A novel imidazole derivative or a pharmaceutically acceptable salt thereof according to the present invention exhibits excellent targeting and inhibitory activity against c-Jun N-terminal kinase 3 (JNK 3), and a pharmaceutical composition including the derivative can be effectively used for use in preventing and treating a degenerative brain disease, such as Alzheimer's dementia.
FIG. 1 is a graph illustrating the inhibitory activity (%) when each of 50 types of protein kinases is treated with 22b (1 μM).
Hereinafter, the present invention will be described in detail.
The present invention provides a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
In Formula 1,
Here, “chiral” is also called a chirality, a hand symmetry, an enantiomer or an optical isomer. Enantiomers are mirror images of each other.
Here, “alkyl” generally refers to a linear or branched saturated hydrocarbon group having a specified number of carbon atoms (e.g., 1 to 12 carbon atoms). Examples of alkyl groups include, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, and n-heptyl. “Alkyl” may be attached to a parent group or a substrate at any ring atom provided that the attachment does not violate valence requirements. Likewise, an alkyl group or alkenyl group may include one or more non-hydrogen substituents provided that the attachment does not violate valence requirements.
Here, “alkoxy” refers to alkyl-O—, wherein the alkyl has been defined above. Examples of alkoxy groups include methoxy and ethoxy without limitation. “Alkoxy” may be attached to a parent group or a substrate at any ring atom provided that the attachment does not violate valence requirements. Likewise, the alkoxy group may include one or more non-hydrogen substituents provided that the attachment does not violate valence requirements.
Here, “halogen” encompasses elements belonging to Group 17 of the periodic table, including fluorine, chlorine, bromine, and iodine.
In one embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 2 below.
In Formula 2,
In another embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 3 below.
In Formula 3,
According to still another embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 4 below.
In Formula 4,
According to yet another embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 5 below.
In Formula 5,
According to yet another embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 6 below.
In Formula 6,
According to yet another embodiment of the present invention, the compound of Formula 1 may be a compound with the structure of Formula 7 below.
In Formula 7,
According to yet another embodiment of the present invention, the imidazole compound of Formula 1 may be
The present invention provides a pharmaceutical composition for use in preventing or treating a degenerative brain disease, which includes the derivative of Formula 1 or a pharmaceutically acceptable salt thereof.
According to yet another embodiment of the present invention, the degenerative brain disease may be Alzheimer's dementia, or Parkinson's disease.
According to yet another embodiment of the present invention, the composition may inhibit JNK3 activity.
Meanwhile, the compound of the present invention may be used in the form of a pharmaceutically acceptable salt, and as a salt, an acid addition salt formed by an acceptable free acid is useful.
The term “salt” used herein is an acid addition salt formed by a pharmaceutically acceptable free acid. The acid addition salt is obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, aliphatic mono- and di-carboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkane dioates, and non-toxic organic acids such as aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically non-toxic salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphate chlorides, bromides, iodides, fluorides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caprates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexane-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxylbenzoates, methoxybenzoates, phthalates, terephthalates, benzenesulfonates, toluenesulfonates, chlorobenzenesulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, β-hydroxylbutyrates, glycolates, malates, tartrates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.
The acid addition salt according to the present invention may be prepared by a conventional method, for example, dissolving a compound in an excess amount of acid aqueous solution, and precipitating the salt using a water-miscible organic solvent, such as methanol, ethanol, acetone, or acetonitrile. In addition, the acid addition salt according to the present invention may be prepared by evaporating a solvent or excess acid from the above mixture and then drying it, or by suction-filtering the precipitated salt.
In addition, a pharmaceutically acceptable metal salt may be prepared using a base. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering an insoluble compound salt, and evaporating and drying the filtrate. Here, it is pharmaceutically suitable to prepare a sodium, potassium, or calcium salt as a metal salt. The corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).
In addition, the compound of the present invention includes not only a pharmaceutically acceptable salt but any salt, isomer, hydrate, and solvate that can be prepared by a conventional method.
As can be confirmed in the following examples, the compound of Formula 1 may be used as a JNK3 inhibitor, and as described in the background of the invention, it is widely known to those of ordinary skill in the art that the JNK3 inhibitor can be used to treat a degenerative brain disease.
The present invention provides a pharmaceutical composition for use in preventing or treating a degenerative brain disease, including the imidazole derivative of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and more particularly, a pharmaceutical composition for use in preventing or treating a degenerative brain disease, uses of the imidazole derivative of Formula 1 or a pharmaceutically acceptable salt thereof for treating the disease, and a method of treating the disease, which includes administering a therapeutically effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt thereof to a subject.
The term “prevention” used herein refers to all actions of inhibiting a degenerative brain disease or delaying the onset thereof by the administration of the pharmaceutical composition according to the present invention.
The term “treatment” used herein refers to all actions involved in alleviating or beneficially changing symptoms of a degenerative brain disease by the administration of the pharmaceutical composition according to the present invention.
The pharmaceutical composition according to the present invention may include a pharmaceutically acceptable carrier as well as an active ingredient. Here, the pharmaceutically acceptable carrier is conventionally used in drug formulation, and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. As well as the above components, the pharmaceutical composition according to the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifier, a suspending agent, or a preservative.
The pharmaceutical composition of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally or topically), and preferably, intracerebrally, according to a desired method, and a dose of the pharmaceutical composition of the present invention may be suitably selected according to a patient's condition and body weight, severity of a disease, a dosage form, an administration route and duration by those of ordinary skill in the art.
The composition according to the present invention is administered at a pharmaceutically effective amount. The “pharmaceutically effective amount” used herein refers to an amount sufficient for treating a disease at a reasonable benefit/risk ratio applicable for medical treatment, and an effective dosage may be determined by parameters including the type and severity of a patient's disease, drug activity, sensitivity to a drug, administration time, an administration route and an excretion rate, the duration of treatment and drugs simultaneously used, and other parameters well known in the medical field. The composition according to the present invention may be administered separately or in combination with other therapeutic agents, and may be sequentially or simultaneously administered with a conventional therapeutic agent, or administered in a single or multiple dose(s). In consideration of all of the above-mentioned parameters, it is important to achieve the maximum effect with the minimum dose without side effects, and such a dose may be easily determined by one of ordinary skill in the art.
Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on a patient's age, sex, condition and body weight, an absorption rate of the active ingredient in the body, an inactivation rate, an excretion rate, a type of disease, or a drug used in combination, and may be generally administered at 0.0001 to 1000 mg/kg, and preferably, 0.001 to 500 mg/kg of body weight daily or every other day, or divided into one or three daily administrations. However, the effective amount may vary depending on an administration route, the severity of obesity, sex, body weight or age, and therefore, the scope of the present invention is not limited by the dose in any way.
The term “individual” used herein refers to a subject in need of treatment, and more specifically, a mammal such as a human or a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow.
Hereinafter, preferred preparation examples and examples are presented to better understand the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following preparation examples.
Compound 3 (2.5 g, 17.85 mmol) was dissolved in a solvent (28.5 mL) in which H2O, THF, and t-BuOH were mixed in a volume ratio of 10:3:5, N-methylmorpholine N-oxide (NMO; 4.8 g, 35.7 mmol) was added and then OsO4 (2 mL, 0.035 mmol) was added, and the resulting mixture was stirred at room temperature for 2 hours. After confirming the termination of the reaction, the resulting mixture was extracted with ethyl acetate (EA), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, Compound 4 (2.9 g, 93%) was obtained.
4-trans (major 2.18 g)1H NMR (400 MHz, DMSO) δ 4.46 (d, J=4.4 Hz, 2H), 4.03 (q, J=7.1 Hz, 3H), 3.90-3.84 (m, 2H), 2.94 (tt, J=9.6, 6.7 Hz, 1H), 1.86-1.74 (m, 4H), 1.16 (t, J=7.1, 0.6 Hz, 3H).
4-cis (minor 0.73 g)1H NMR (400 MHz, DMSO) δ 4.37 (d, J=4.5 Hz, 2H), 4.03 (q, J=7.1 Hz, 2H), 3.78-3.72 (m, 2H), 2.72-2.60 (tt, 1H), 1.98-1.90 (m, 4H), 1.24 (t, J=8.4, 5.8 Hz, 3H). HRMS (ESI+) calculated for C8H15O4 [M+H]+: 175.0965, found 175.2584.
Compound 4 (2.9 g, 16.9 mmol), which had been obtained in the previous experiment, was dissolved in MC (85 mL), TEA (5.1 mL, 37.1 mmol) was added at 0° C., and then the resulting mixture was stirred for 10 minutes. Afterward, methanesulfonic acid (MsCl; 2.85 mL, 37.1 mmol) was added and then stirring was performed at room temperature for 2 hours. After confirming the termination of the reaction, the resulting product was extracted with ethyl acetate (EA), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, Compound 4′(5.58 g) was obtained.
4′-trans (major 4.19 g)1H NMR (400 MHz, CDCl3) δ 5.10 (p, J=3.7 Hz, 2H), 4.21-4.11 (q, 2H), 3.29-3.14 (m, 1H), 3.10 (s, 6H), 2.41-2.28 (m, 4H), 1.29-1.23 (t, 3H).
4′-cis (minor 1.39 g)1H NMR (400 MHz, CDCl3) δ 5.03-4.97 (m, 2H), 4.11-3.99 (q, 2H), 3.67 (s, 6H), 2.93-2.86 (m, 1H), 2.51-2.42 (m, 4H), 1.33 (t, J=7.1 Hz, 3H).
Compound 4 (2.9 g, 16.9 mmol), which had been obtained in the previous experiment, was dissolved in DMF (85 mL), sodium azide (NaN3; 5.5 g, 85 mmol) was added, and then the resulting mixture was stirred at 80° C. for 24 hours. After confirming the termination of the reaction, the resulting product was extracted with ethyl acetate (EA), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using HEX and EA (HEX:EA=10:1) as mobile phase, thereby obtaining Compound 5.
5-trans (minor 0.62 g)1H NMR (400 MHz, CDCl3) δ 4.14 (q, J=7.1 Hz, 2H), 4.00 (p, J=4.2 Hz, 2H), 3.10 (tt, J=9.8, 6.2 Hz, 1H), 2.22-2.07 (m, 8H), 1.26 (t, J=7.1 Hz, 3H).
5-cis (major 1.85 g) 1H NMR (400 MHz, CDCl3) δ 4.18 (q, J=7.1 Hz, 2H), 3.89-3.83 (m, 2H), 2.87 (p, J=8.4 Hz, 1H), 2.27-2.20 (m, 4H), 1.27 (t, J=7.1 Hz, 3H).
Compound 5 (710 mg, 3.17 mmol), which had been obtained in the previous experiment, was dissolved in MeOH (12 mL), and then 20% Pd(OH)2 on carbon (220 mg, 0.317 mmol) was added. The reaction vessel was substituted with hydrogen gas and stirred at room temperature for 1 hour. After confirming the termination of the reaction, Pd(OH)2 was removed by filtration using Celite with ethyl acetate as a solvent, and the solvent was concentrated, thereby obtaining Compound 6 (533 mg, 97%).
6-trans (minor 0.14 g)1H NMR (400 MHz, CDCl3) δ 4.15-4.09 (q, 2H), 3.47-3.41 (m, 2H), 3.12 (td, J=9.6, 4.5 Hz, 1H), 2.20-2.11 (m, 2H), 1.96-1.87 (m, 2H), 1.27-1.22 (t, 3H).
6-cis (major 0.40 g)1H NMR (400 MHz, CDCl3) δ 4.14 (q, J=11.0, 4.0 Hz, 2H), 3.28 (dd, J=7.7, 3.2 Hz, 2H), 2.89-2.72 (m, 1H), 2.20 (dd, J=9.1, 6.2 Hz, 2H), 1.89-1.77 (m, 2H), 1.28-1.23 (t, 3H).
Compound 6 (1.119 g, 6.50 mmol), which had been obtained in the previous experiment, was dissolved in EtOH, and then ethyl 2-naphthimidate was added. The resulting mixture was stirred at 80° C. for 12 hours. After confirming the termination of the reaction, the resulting product was extracted with ethyl acetate (EA), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using MC, MeOH, and NH4OH (MC:MeOH:NH4OH=100:10:1) as a mobile phase, thereby obtaining Compound 8a (1.91 g, 85%).
8a (1.9062 g, 85%). 1H NMR (400 MHz, DMSO-d6) δ 8.19-8.12 (m, 2H), 7.63-7.56 (m, 1H), 4.50 (d, J=1.7 Hz, 2H), 4.04 (td, J=7.1, 1.7 Hz, 2H), 3.41-3.23 (m, 1H), 2.68-2.57 (m, 1H), 1.99-1.84 (m, 4H), 1.19-1.13 (m, 3H). HRMS (ESI+) calculated for C16H17F4N2O2 [M+H]+: 345.1221, found 345.2313.
8b (1005 mg, 53%)1H NMR (400 MHz, DMSO) δ 8.00 (d, J=1.9 Hz, 1H), 7.77 (dd, J=8.4, 1.9 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 4.49 (s, 2H), 4.04 (q, J=7.1 Hz, 2H), 3.59 (s, 1H), 2.67-2.56 (m, 1H), 1.99-1.84 (m, 4H), 1.16 (t, J=7.1 Hz, 3H); MS(ESI+) calculated for C15H17C12N2O2+ [M+H]+: 327.0662, found 327.0825.
8c (2100 mg, 69%). 1H NMR (400 MHz, DMSO-D6) δ 8.32 (s, 1H), 7.98-7.91 (m, 4H), 7.61-7.54 (m, 2H), 7.40 (d, J=47.8 Hz, 1H), 4.53 (s, 2H), 4.05 (q, J=7.1 Hz, 2H), 2.64 (dt, J=17.9, 5.9 Hz, 1H), 2.04-1.96 (m, 2H), 1.91 (td, J=12.5, 6.5 Hz, 2H), 1.19-1.13 (t, 3H). 1H NMR (400 MHz, DMSO) δ 8.32 (s, 1H), 7.97-7.92 (m, 4H), 7.58-7.55 (m, 2H), 4.35 (s, 2H), 3.90 (q, J=7.1 Hz, 2H), 2.73 (t, J=7.4 Hz, 1H), 2.27 (dt, J=12.9, 7.3 Hz, 2H), 1.95-1.87 (m, 2H), 1.05 (t, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C19H20N2O2 [M+H]+: 308.3810, found 309.2216.
8d (601 mg, 47%). 1H NMR (400 MHz, Acetone) δ 7.45-7.40 (m, 2H), 6.85 (dd, J=8.5, 3.6 Hz, 1H), 6.04 (d, J=1.7 Hz, 2H), 4.56 (dd, J=4.4, 2.2 Hz, 1H), 4.38 (dd, J=6.1, 2.9 Hz, 1H), 4.07 (q, J=8.5, 5.7 Hz, 2H), 2.77-2.71 (m, 1H), 2.33-2.23 (m, 2H), 1.99-1.91 (m, 2H), 1.19 (t, J=7.6, 6.7 Hz, 3H); 1H NMR (400 MHz, Acetone) δ 7.45-7.40 (m, 2H), 6.85 (dd, J=8.5, 3.6 Hz, 1H), 6.04 (d, J=1.7 Hz, 2H), 4.56 (dd, J=4.4, 2.2 Hz, 1H), 4.38 (dd, J=6.1, 2.9 Hz, 1H), 4.00-3.94 (q, 2H), 2.77-2.71 (m, 1H), 2.33-2.23 (m, 2H), 1.99-1.91 (m, 2H), 1.13 (t, J=7.1 Hz, 3H); HRMS (ESI+) calculated for C16H18N2O4 [M+H]+: 302.3300, found 303.2079.
8e (590 mg, 46%)1H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 7.94 (s, 1H), 7.86 (dd, J=8.5, 2.0 Hz, 1H), 6.99 (d, J=8.5 Hz, 1H), 4.68 (dd, J=10.5, 7.1 Hz, 4H), 3.25 (t, J=8.7 Hz, 2H), 3.07 (m, J=4.8 Hz, 1H), 2.31 (dd, J=14.0, 3.0 Hz, 2H), 2.22-2.14 (m, 2H), 1.20 (t, 3H); 1H NMR (400 MHz, DMSO-d6) δ 10.80 (s, 1H), 7.94 (s, 1H), 7.86 (dd, J=8.5, 2.0 Hz, 1H), 6.99 (d, J=8.5 Hz, 1H), 4.68 (dd, J=10.5, 7.1 Hz, 4H), 4.08 (q, J=7.1 Hz, 1H), 3.25 (t, J=8.7 Hz, 2H), 3.07 (d, J=4.8 Hz, 1H), 2.31 (dd, J=14.0, 3.0 Hz, 2H), 2.22-2.14 (m, 2H), 1.03 (t, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C17H21N2O3 [M+H]+: 301.1547, found 301.2634.
Oxalyl chloride (519 μL, 6.05 mmol) was dissolved in MC (15 mL), and stirred at −78° C. for approximately 10 minutes. Dimethyl sulfoxide (DMSO; 780 μL, 11 mmol) was dissolved in MC (15 mL), and then added to the oxalyl chloride being stirred, followed by stirring for 15 minutes. Compound 8a (1.89 g, 5.5 mmol), which had been obtained in the previous experiment, was dissolved in MC (25 mL) and added dropwise to a reaction vessel while stirring, and then further stirred for 30 minutes. Afterward, triethyl amine (TEA; 3.2 mL, 27.5 mmol) was added dropwise. After stirring for 10 minutes, the resulting solution was brought to room temperature to proceed with the reaction. After confirming the completion of the reaction, the resulting product was extracted with ethyl acetate (EA), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using HEX and EA (HEX:EA=1:2) as mobile phase, thereby obtaining Compound 9.
9a (1.243 g, 66%). 1H NMR (400 MHz, DMSO-d6) δ 12.57 (s, 1H), 8.21 (dd, J=6.9, 2.0 Hz, 1H), 8.19-8.14 (m, 1H), 7.56 (dd, J=10.3, 9.1 Hz, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.79 (tt, J=9.1, 6.1 Hz, 1H), 3.09-2.85 (m, 3H), 2.78 (dd, J=14.5, 5.6 Hz, 1H), 1.22 (t, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C16H18F4N2O2 [M+H]+: 343.2148, found 343.2148.
9b (209 mg, 22%)1H NMR (400 MHz, DMSO) δ 12.53 (s, 1H), 8.06 (d, J=2.0 Hz, 1H), 7.82 (dd, J=8.5, 2.1 Hz, 1H), 7.67 (d, J=8.5 Hz, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.79 (ddd, J=9.2, 7.7, 4.6 Hz, 1H), 3.08-2.85 (m, 4H), 1.22 (t, J=7.1 Hz, 3H). MS(ESI+) calculated for C15H15C12N2O2+ [M+H]+: 325.0505, found 325.4621.
9c (1030 mg, 41%). 1H NMR (400 MHz, DMSO) δ 12.50 (s, 1H), 8.35 (s, 1H), 8.04 (dd, J=8.6, 1.7 Hz, 1H), 7.94 (s, 1H), 7.89 (d, J=8.8 Hz, 2H), 7.55-7.47 (m, 2H), 4.14 (q, J=6.9 Hz, 2H), 3.81 (ddd, J=9.1, 7.6, 4.6 Hz, 1H), 3.00 (dd, J=14.4, 9.2 Hz, 2H), 2.90 (dd, J=13.9, 6.1 Hz, 2H), 1.23 (t, J=7.1 Hz, 3H); 13C NMR (101 MHz, DMSO-D6) δ 175.03 (s), 174.58 (s), 149.25 (s), 133.06 (s), 132.36 (s), 128.99 (s), 128.20 (s), 127.89 (s), 127.66 (s), 126.59 (s), 125.94 (s), 122.97 (s), 122.36 (s), 60.31 (s), 51.80 (s), 45.89 (d, J=11.3 Hz), 27.40 (s), 14.12 (s); HRMS (ESI+) calculated for C19H18N2O2 [M+H]+: 306.3650, found 307.2050.
9d (717 mg, 58%). 1H NMR (400 MHz, Acetone) δ 7.42 (dd, J=7.4, 1.6 Hz, 2H), 6.89-6.85 (m, 1H), 6.02 (s, 2H), 4.16 (q, J=7.1 Hz, 2H), 3.74 (dd, J=15.2, 8.0 Hz, 1H), 3.01-2.83 (m, 4H), 1.25 (t, J=7.1 Hz, 3H). 13C NMR (101 MHz, DMSO-D6) δ 174.60 (s), 149.13 (s), 147.59 (s), 146.82 (s), 142.62 (s), 137.14 (s), 125.94 (s), 118.06 (s), 108.50 (s), 104.79 (s), 101.11 (s), 60.28 (s), 45.90 (s), 27.60 (s), 27.14 (s), 14.12 (s); HRMS (ESI+) calculated for C16H16N2O4 [M+H]+: 300.3140, found 301.1913.
9e (991 mg, 83%)1H NMR (400 MHz, Acetone) 67.78 (d, J=1.3 Hz, 1H), 7.68-7.64 (m, 1H), 6.73 (d, J=8.4 Hz, 1H), 4.57 (t, J=11.5, 5.9 Hz, 2H), 4.16 (q, J=7.1 Hz, 2H), 3.75 (dt, J=15.3, 7.6 Hz, 1H), 3.22 (t, J=8.7 Hz, 2H), 2.90 (d, J=44.8 Hz, 4H), 1.25 (t, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C17H19N2O3 [M+H]+: 299.1390, found 299.2108.
Compound 9a (268 mg, 0.78 mmol), which had been obtained in the previous experiment, was dissolved in anhydrous DMF (3.9 mL, 0.2M), and CsCO3 (263 mg, 0.81 mmol) was added. 4-Chloro-2-(methylthio)pyrimidine (91 μL, 0.78 mmol) was added, and then stirred in a reactor microwave (Biotage) for 1 hour at 80° C. After completing the reaction, an organic layer was extracted using ethyl acetate (EtOAc), and washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and evaporating the solvent, separation and purification were performed through column chromatography (MC:MeOH=40:1), thereby obtaining Compound 10a (179 mg, 49%).
1H NMR (400 MHz, DMSO) δ 8.65 (d, J=5.4 Hz, 1H), 7.81 (dd, J=6.8, 1.7 Hz, 1H), 7.72 (dd, J=5.5, 3.1 Hz, 1H), 7.56-7.48 (m, 1H), 7.07 (d, J=5.4 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 3.86 (tt, J=9.2, 6.1 Hz, 1H), 3.24 (ddd, J=21.7, 15.5, 7.4 Hz, 2H), 3.00 (dd, J=14.8, 9.5 Hz, 1H), 2.89 (dd, J=15.0, 5.9 Hz, 1H), 2.16 (s, 3H), 1.22 (t, J=7.1 Hz, 3H); HIRMS (ESI+) calculated for C21H19F4N4O2S [M+H]+: 467.1159, found 467.3361.
10b (135 mg, 49%)1H NMR (400 MHz, DMSO) δ 8.66 (d, J=5.4 Hz, 1H), 7.71 (d, J=2.0 Hz, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.33 (dd, J=8.4, 2.1 Hz, 1H), 7.04 (d, J=5.4 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 3.86 (tt, J=9.2, 6.0 Hz, 1H), 3.25 (d, J=9.9 Hz, 1H), 3.18 (dd, J=15.8, 5.6 Hz, 1H), 3.00 (dd, J=15.5, 8.9 Hz, 1H), 2.89 (dd, J=15.0, 5.9 Hz, 1H), 2.22 (s, 3H), 1.22 (s, 3H); MS(ESI+) calculated for C20H19Cl2N4O2S+ [M+H]+: 449.0600, found 449.7283.
10c (717 mg, 59%). 1H NMR (400 MHz, Acetone) δ 8.48 (d, J=5.4 Hz, 1H), 8.06 (d, J=0.9 Hz, 1H), 7.95-7.89 (m, 3H), 7.58-7.53 (m, 3H), 6.81 (d, J=5.4 Hz, 1H), 4.19 (q, J=7.1, 4.8 Hz, 2H), 3.87 (tt, J=8.9, 6.4 Hz, 1H), 3.40-3.32 (m, 2H), 3.07-2.99 (m, 2H), 2.28 (s, 3H), 1.30-1.26 (t, 3H). 13C NMR (101 MHz, DMSO-D6) δ 174.21 (s), 172.18 (s), 159.60 (s), 156.21 (s), 149.85 (s), 144.82 (s), 133.63 (s), 132.61 (d, J=4.0 Hz), 128.74 (s), 128.20 (s), 127.77 (s), 127.55 (s), 126.83 (s), 126.57 (s), 126.03 (s), 109.21 (s), 59.72 (s), 45.16 (s), 28.53 (s), 28.13 (s), 14.05 (d, J=4.2 Hz). IRMS (ESI+) calculated for C24H22N4O2S [M+H]+: 430.5260, found 431.7963.
10d (713 mg, 72%). 1H NMR (400 MHz, MeOD) δ 8.46 (d, J=5.5 Hz, 1H), 6.92-6.87 (m, 3H), 6.73 (d, J=5.5 Hz, 1H), 6.03 (s, 2H), 4.22 (q, J=7.1 Hz, 2H), 3.86 (ddd, J=12.8, 7.5, 4.3 Hz, 1H), 3.32-3.29 (m, 2H), 3.08-2.99 (m, 2H), 1.33-1.29 (t, 3H). 13C NMR (101 MHz, DMSO-D6) δ 174.27 (s), 172.15 (s), 159.51 (s), 156.24 (s), 149.59 (s), 147.84 (s), 147.29 (s), 144.21 (s), 133.07 (s), 125.11 (s), 122.96 (s), 109.31 (s), 108.86 (s), 108.33 (s), 101.43 (s), 60.43 (s), 45.07 (s), 28.57 (s), 28.13 (s), 14.09 (s), 13.26 (s). HIRMS (ESI+) calculated for C21H20N4O4S [M+H]+: 424.4750, found 425.7469.
10e (12 mg, 17%)1H NMR (400 MHz, DMSO-d6) δ 8.54 (d, J=5.5 Hz, 1H), 7.29 (d, J=1.0 Hz, 1H), 7.07 (dd, J=8.3, 1.8 Hz, 1H), 6.74 (dd, J=6.8, 5.4 Hz, 2H), 4.56 (t, J=8.7 Hz, 2H), 4.14 (q, J=7.1 Hz, 2H), 3.81 (tt, J=9.2, 6.0 Hz, 1H), 3.29-3.21 (m, 1H), 3.16 (dd, J=11.7, 6.0 Hz, 3H), 2.96 (dd, J=14.7, 9.4 Hz, 1H), 2.86 (dd, J=14.9, 5.8 Hz, 1H), 2.34 (s, 3H), 1.22 (t, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C22H23N4O3S [M+H]+: 423.1485, found 423.2263.
Compound 10a (364 mg, 0.78 mmol), which had been obtained in the previous experiment, and potassium peroxomonosulfate were mixed in a solvent in which methanol and water (1:1) were mixed (15.6 mL), and the resulting mixture was stirred at room temperature for 1 hour. After the starting material completely disappeared in TLC, the methanol was concentrated in a vacuum. The concentrated mixture was diluted by adding water, stirred until a solid product was separated, and then filtrated, thereby obtaining the solid product (Compound 11a; 136.8 mg, 0.27 mmol).
11a (136.8 mg, 35%). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J=14.8, 5.9 Hz, 1H), 7.79-7.62 (m, 2H), 7.58-7.47 (m, 1H), 6.37 (dd, J=42.0, 36.8 Hz, 1H), 4.19-4.06 (m, 2H), 3.88-3.79 (m, 1H), 3.32 (s, 3H), 3.23-3.16 (m, 1H), 3.14-3.03 (m, 1H), 2.98 (dd, J=14.8, 8.9 Hz, 1H), 2.87 (dd, J=16.1, 6.2 Hz, 1H), 1.21 (dd, J=5.3, 3.0 Hz, 3H). HRMS (ESI+) calculated for C21H19F4N4O4S [M+H]+: 499.1058, found 499.3828.
11b (35 mg, 38%)1H NMR (400 MHz, DMSO) δ 8.66 (d, J=5.4 Hz, 1H), 7.71 (d, J=2.0 Hz, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.33 (dd, J=8.4, 2.1 Hz, 1H), 7.04 (d, J=5.4 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 3.86 (tt, J=9.2, 6.0 Hz, 1H), 3.22 (s, 3H), 3.25 (d, J=9.9 Hz, 1H), 3.18 (dd, J=15.8, 5.6 Hz, 1H), 3.00 (dd, J=15.5, 8.9 Hz, 1H), 2.89 (dd, J=15.0, 5.9 Hz, 1H), 2.22 (s, 3H); MS(ESI+) calculated for C20H19Cl2N4O4SV[M+H]+: 481.0499, found 481.8831.
11c (43 mg, 56%). 1H NMR (400 MHz, DMSO-D6) δ 8.23 (d, J=5.3 Hz, 1H), 8.00 (s, 1H), 7.90 (d, J=8.6 Hz, 3H), 7.60-7.50 (m, 3H), 7.47-7.41 (m, 1H), 6.27 (s, 1H), 4.15 (q, J=7.0 Hz, 2H), 3.85 (dt, J=9.1, 7.7 Hz, 2H), 3.76 (s, 1H), 3.21 (dt, J=15.6, 8.8 Hz, 2H), 2.96 (ddd, J=20.1, 14.7, 7.4 Hz, 4H), 2.74-2.55 (m, 2H), 1.52 (d, J=49.5 Hz, 2H), 1.23 (t, J=7.1 Hz, 3H).
HRMS (ESI+) calculated for C24H22N4O4S [M+H]+: 462.5240, found 463.8073.
11d (250 mg, 82%). 1H NMR (400 MHz, DMSO-d6) δ 8.98 (d, J=5.6 Hz, 1H), 7.31 (d, J=5.6 Hz, 1H), 7.05 (dd, J=1.5, 0.5 Hz, 1H), 6.97-6.90 (m, 2H), 6.08 (s, 2H), 4.14 (q, J=7.1 Hz, 2H), 3.88-3.80 (m, 1H), 3.30 (dd, J=12.0, 2.5 Hz, 1H), 3.28 (s, 3H), 3.26-3.17 (m, 1H), 3.03-2.81 (m, 2H), 1.22 (t, J=7.1 Hz, 3H); HRMS (ESI+) calculated for C21H20N4O6S [M+H]+: 456.4730, found 457.2179.
11e (240 mg, 74%). 1H NMR (400 MHz, DMSO-d6) δ 8.97 (d, J=5.6 Hz, 1H), 7.34 (s, 1H), 7.30 (d, J=5.6 Hz, 1H), 7.17 (dd, J=8.3, 2.0 Hz, 1H), 6.79 (d, J=8.3 Hz, 1H), 4.58 (t, J=8.8 Hz, 2H), 4.14 (q, J=7.1 Hz, 2H), 3.86-3.82 (m, 1H), 3.24 (s, 3H), 3.17 (s, 2H), 3.01 (d, J=9.1 Hz, 1H), 2.97 (d, J=10.3 Hz, 1H), 2.91-2.83 (m, 2H), 1.23 (d, J=7.1 Hz, 3H). HRMS (ESI+) calculated for C22H23N4O5S [M+H]+: 455.1384, found 455.2014.
Compound 11 that had been obtained in the previous reaction was dissolved in THF (1.5 mL), tert-butyl 3-aminopyrrolidine-1-carboxylate (*=S, n=108.5 μL, 0.60 mmol) was added, and then the resulting mixture was stirred at 60° C. for 22 hours. After confirming the termination of the reaction, the resulting product was extracted with ethyl acetate (EtOAc), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using MC and MeOH (MC:MeOH=40:1) as mobile phase, thereby obtaining Compound 12a (25 mg, 40%).
12a (81 mg, 89%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J=5.0 Hz, 1H), 7.76 (t, J=23.6 Hz, 3H), 7.50 (t, J=9.2 Hz, 1H), 6.48 (s, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.88-3.79 (m, 1H), 3.29-3.08 (m, 5H), 2.98 (dd, J=14.3, 9.6 Hz, 2H), 2.88 (dd, J=14.8, 5.5 Hz, 1H), 1.79 (d, J=7.7 Hz, 1H), 1.61 (dd, J=21.3, 13.5 Hz, 1H), 1.38 (s, 9H), 1.22 (t, J=7.0 Hz, 4H). HRMS (ESI+) calculated for C30H35F4N6O4 [M+H]+: 619.2650, found 619.3580.
12b (21 mg, 85%)1H NMR (400 MHz, DMSO) δ 8.35 (d, J=5.1 Hz, 1H), 7.80 (s, 1H), 7.69-7.57 (m, 2H), 7.31 (d, J=8.2 Hz, 1H), 6.45 (s, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.87-3.76 (m, 1H), 3.34 (s, 1H), 3.25-2.81 (m, 8H), 1.85-1.75 (m, 1H), 1.57-1.49 (m, 1H), 1.39 (d, J=4.3 Hz, 9H), 1.23 (d, J=7.0 Hz, 3H). MS(ESI+) calculated for C28H33C12N6O4+ [M+H]+: 587.1935, found 587.8881.
12c (22 mg, 52%). 1H NMR (400 MHz, DMSO-D6) δ 8.24 (d, J=5.3 Hz, 1H), 8.00 (s, 1H), 7.93-7.87 (m, 3H), 7.71 (s, 1H), 7.56-7.49 (m, 2H), 7.44 (d, J=8.4 Hz, 1H), 6.39-6.11 (m, 1H), 4.15 (q, J=7.1 Hz, 2H), 3.91-3.80 (m, 1H), 3.76-3.51 (m, 1H), 3.30-3.09 (m, 4H), 3.09-2.72 (m, 4H), 1.90-1.48 (m, 2H), 1.38 (d, J=6.2 Hz, 9H), 1.23 (t, J=7.1 Hz, 3H). 13C NMR (101 MHz, DMSO-D6) δ 174.33 (s), 162.07 161.52 (m), 156.67 (s), 155.05 (s), 153.40 (s), 144.31 (s), 132.55 (d, J=4.9 Hz), 129.63 (s), 128.22 (s), 127.84 (d, J=59.2 Hz), 126.64 (d, J=17.5 Hz), 126.17 (s), 100.94 (s), 78.17 (s), 60.42 (s), 50.73 (s), 50.33 (s), 45.18 (s), 43.63 (s), 30.61 (s), 29.79 (s), 28.45 (s), 28.15 (s), 14.10 (s). HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 568.6780, found 569.9207.
12d (61 mg, 49%). 1H NMR (400 MHz, DMSO-D6) δ 8.26 (d, J=5.2 Hz, 1H), 7.77 (s, 1H), 6.96-6.87 (m, 2H), 6.83 (dd, J=8.1, 1.3 Hz, 1H), 6.19 (d, J=32.4 Hz, 1H), 6.05 (s, 2H), 4.13 (q, J=7.1 Hz, 2H), 3.98 (t, J=29.9 Hz, 1H), 3.86-3.72 (m, 1H), 3.53-3.37 (m, 2H), 3.26-3.08 (m, 4H), 3.00-2.78 (m, 2H), 2.09-1.72 (m, 2H), 1.39 (d, J=4.4 Hz, 9H), 1.21 (t, J=7.1 Hz, 3H). 3C NMR (101 MHz, DMSO-D6) δ 174.37 (s), 162.29 (s), 161.98 (s), 155.74 (s), 153.58 (s), 147.65 (s), 147.19 (s), 143.72 (s), 133.15 (s), 131.68 (s), 122.72 (s), 108.72 (s), 108.18 (s), 102.48 (s), 101.40 (s), 78.25 (s), 60.40 (s), 54.89 (s), 51.00 (s), 45.12 (s), 43.87 (s), 30.71 (s), 29.88 (s), 28.52 (s), 28.06 (d, J=19.3 Hz), 14.09 (s). HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 562.6270, found 563.9796.
12e (43 mg, 63%). 1H NMR (400 MHz, DMSO) δ 8.27 (dd, J=11.8, 5.3 Hz, 1H), 7.79 (d, J=22.4 Hz, 1H), 7.29 (s, 1H), 7.05 (d, J=8.2 Hz, 1H), 6.77-6.72 (m, 1H), 6.25 (d, J=34.8 Hz, 1H), 4.54 (dd, J=8.5, 6.1 Hz, 2H), 4.15-4.10 (m, 2H), 3.80 (s, 2H), 3.46 (d, J=5.4 Hz, 1H), 3.28-3.08 (m, 6H), 2.99-2.90 (m, 1H), 2.84 (dd, J=14.5, 5.5 Hz, 1H), 1.99-1.90 (m, 1H), 1.75 (dt, J=12.4, 6.2 Hz, 1H), 1.48 (ddd, J=12.6, 5.9, 3.8 Hz, 1H), 1.21 (dd, J=7.0, 1.3 Hz, 3H), 0.81 (t, J=2.7 Hz, 1H), 0.77 (dd, J=4.9, 2.1 Hz, 2H), 0.71 (d, J=2.0 Hz, 2H); HRMS (ESI+) calculated for C30H37N6O5 [M+H]+: 561.2820, found 561.3150.
13a (79 mg, 87%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J=5.0 Hz, 1H), 7.76 (t, J=23.7 Hz, 3H), 7.50 (t, J=9.2 Hz, 1H), 6.48 (s, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.83 (dd, J=13.4, 7.2 Hz, 1H), 3.28-3.06 (m, 5H), 2.99 (dd, J=14.4, 9.6 Hz, 2H), 2.88 (dd, J=14.9, 5.4 Hz, 1H), 1.79 (dd, J=6.6, 5.1 Hz, 1H), 1.62 (dd, J=17.4, 9.7 Hz, 1H), 1.38 (s, 9H), 1.21 (dd, J=13.7, 6.6 Hz, 4H). HRMS (ESI+) calculated for C30H35F4N6O4 [M+H]+: 619.2650, found 619.4301.
13b (41 mg, 84%)1H NMR (400 MHz, DMSO) δ 8.35 (d, J=5.2 Hz, 1H), 7.80 (s, 1H), 7.69-7.59 (m, 2H), 7.31 (d, J=8.3 Hz, 1H), 6.45 (s, 1H), 4.13 (d, J=7.1 Hz, 2H), 3.88-3.78 (m, 1H), 3.36 (s, 1H), 3.29-2.79 (m, 8H), 1.85-1.75 (m, 1H), 1.62-1.46 (m, 1H), 1.39 (d, J=4.7 Hz, 9H), 1.21 (d, J=7.0 Hz, 3H). MS(ESI+) calculated for C28H33Cl2N6O4+ [M+H]+: 587.1935, found 587.9601.
13c (24 mg, 71%). 1H NMR (400 MHz, DMSO-D6) δ 8.24 (d, J=5.3 Hz, 1H), 8.00 (s, 1H), 7.93-7.91 (m, 1H), 7.89 (d, J=8.4 Hz, 2H), 7.70 (s, 1H), 7.56-7.50 (m, 2H), 7.45 (d, J=8.4 Hz, 1H), 6.38-6.13 (m, 1H), 4.21-4.05 (m, 4H), 3.90-3.80 (m, 1H), 3.74-3.52 (m, 1H), 3.28-3.18 (m, 4H), 3.08-2.96 (m, 2H), 2.90 (dt, J=33.7, 12.3 Hz, 2H), 1.72-1.47 (m, 2H), 1.38 (d, J=6.1 Hz, 9H), 1.23 (t, J=7.1 Hz, 3H); 13C NMR (101 MHz, DMSO-D6) δ 174.33 (s), 162.10-161.70 (m), 153.47 (d, J=11.0 Hz), 144.31 (s), 132.69-132.24 (m), 128.13 (s), 127.54 (s), 126.64 (d, J=18.2 Hz), 126.12 (s), 125.85 (s), 124.56 (s), 102.10 (s), 78.17 (s), 60.42 (s), 50.74 (s), 50.39 (s), 48.59 (s), 45.19 (s), 30.61 (s), 29.78 (s), 28.45 (s), 28.15 (s), 14.11 (s). HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 568.6780, found 569.5607.
13d (109 mg, 89%). 1H NMR (400 MHz, DMSO-D6) δ 8.26 (d, J=5.2 Hz, 1H), 7.77 (s, 1H), 6.96-6.79 (m, 3H), 6.19 (d, J=32.4 Hz, 1H), 6.05 (s, 2H), 4.13 (q, J=7.1 Hz, 2H), 4.09-3.88 (m, 1H), 3.85-3.74 (m, 1H), 3.44 (ddd, J=23.8, 13.6, 6.2 Hz, 2H), 3.30-3.06 (m, 4H), 3.02-2.79 (m, 2H), 2.08-1.72 (m, 2H), 1.39 (d, J=4.4 Hz, 9H), 1.21 (t, J=7.1 Hz, 3H). 13C NMR (101 MHz, DMSO-D6) δ 174.34 (s), 162.12-161.87 (m), 153.53 (d, J=6.2 Hz), 147.65 (s), 147.17 (d, J=0.9 Hz), 143.72 (s), 122.70 (s), 108.71 (s), 108.16 (s), 101.40 (d, J=1.7 Hz), 78.22 (s), 60.39 (s), 54.87 (s), 50.99 (s), 45.12 (s), 44.11 (s), 29.90 (s), 28.53 (s), 28.11 (d, J=6.4 Hz), 14.07 (s). HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 562.6270, found 563.9435.
13e (45 mg, 66%). 1H NMR (400 MHz, DMSO) δ 8.27 (dd, J=12.0, 5.2 Hz, 1H), 7.87-7.71 (m, 1H), 7.29 (s, 1H), 7.05 (d, J=8.2 Hz, 1H), 6.74 (dd, J=8.3, 1.8 Hz, 1H), 6.35-6.06 (m, 1H), 4.59-4.50 (m, 2H), 4.13 (dd, J=14.0, 7.0 Hz, 2H), 3.76 (d, J=26.0 Hz, 2H), 3.46 (d, J=5.4 Hz, 1H), 3.30-3.09 (m, 6H), 3.00-2.90 (m, 1H), 2.84 (dd, J=14.5, 5.1 Hz, 1H), 1.93 (dd, J=18.0, 8.6 Hz, 1H), 1.75 (dt, J=12.8, 6.3 Hz, 1H), 1.49 (tt, J=7.7, 4.8 Hz, 1H), 1.21 (t, J=7.1 Hz, 3H), 0.81 (t, J=2.6 Hz, 1H), 0.79-0.76 (m, 2H), 0.73-0.71 (m, 2H). HRMS (ESI+) calculated for C30H37N6O5 [M+H]+: 561.2820, found 561.2791.
14a (77.5 mg, 83%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J=4.8 Hz, 1H), 7.70 (dd, J=28.3, 6.9 Hz, 2H), 7.56-7.48 (m, 1H), 6.48 (s, 1H), 4.17-4.10 (m, 2H), 3.83 (ddd, J=15.4, 9.2, 6.4 Hz, 1H), 3.61 (d, J=10.3 Hz, 2H), 3.13 (dd, J=32.8, 23.1 Hz, 2H), 3.04-2.94 (m, 2H), 2.93-2.82 (m, 2H), 1.61 (s, 2H), 1.43-1.26 (m, 9H), 1.22 (td, J=7.1, 5.0 Hz, 5H), 1.19-1.13 (m, 1H), 0.88-0.78 (m, 1H). HRMS (ESI+) calculated for C29H33F4N6O4 [M+H]+: 605.2494, found 605.2434.
14b (43 mg, 85%)1H NMR (400 MHz, DMSO-d6) 8.34 (d, J=5.1 Hz, 1H), 7.60 (d, J=8.5 Hz, 2H), 7.53 (s, 1H), 7.30-7.25 (m, 1H), 6.44 (s, 1H), 4.13 (dd, J=9.5, 4.0 Hz, 2H), 3.81 (td, J=9.1, 4.8 Hz, 1H), 3.74-3.43 (m, 2H), 3.23-2.73 (m, 7H), 1.83-1.49 (m, 4H), 1.36 (dd, J=10.5, 6.2 Hz, 9H), 1.24-1.21 (m, 3H); MS(ESI+) calculated for C29H35Cl2N6O4+ [M+H]+: 601.2091, found 601.9667.
14c (49 mg, 90%). 1H NMR (400 MHz, MeOD) δ 8.20 (d, J=5.2 Hz, 1H), 8.00 (s, 1H), 7.90 (dd, J=12.6, 5.6 Hz, 3H), 7.59-7.52 (m, 2H), 7.44 (d, J=8.3 Hz, 1H), 6.35 (s, 1H), 4.28-4.20 (m, 2H), 3.93-3.87 (m, 1H), 3.54 (dd, J=80.1, 58.5 Hz, 3H), 3.28 (s, 1H), 3.07 (ddd, J=31.5, 20.3, 11.9 Hz, 4H), 1.52-1.35 (m, 9H), 1.34-1.30 (m, 6H), 0.91 (d, J=7.0 Hz, 2H); 13C NMR (101 MHz, DMSO-D6) δ 174.29 (s), 161.77 (s), 160.40 (d, J=5.3 Hz), 153.82 (d, J=15.0 Hz), 144.22 (s), 132.69-132.41 (m), 128.18 (d, J=4.2 Hz), 127.55 (s), 126.72 (s), 126.31 (d, J=39.9 Hz), 126.01-125.94 (m), 101.23 (s), 78.44 (s), 60.40 (s), 54.88 (s), 47.49 (s), 46.76 (s), 45.17 (s), 43.71 (s), 28.61-28.55 (m), 28.45 (s), 28.33-27.48 (m), 22.97 (s), 14.10 (s); HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 582.7050, found 583.9634.
14d (102.1 mg, 95%). 1H NMR (400 MHz, DMSO-D6) δ 8.25 (d, J=5.2 Hz, 1H), 7.43 (d, J=52.5 Hz, 1H), 6.84 (dt, J=8.1, 4.8 Hz, 3H), 6.14 (dd, J=25.2, 19.6 Hz, 1H), 6.05 (s, 2H), 4.12 (qd, J=7.0, 3.5 Hz, 2H), 3.84-3.69 (m, 2H), 3.66-3.32 (m, 2H), 3.23-2.57 (m, 6H), 1.96-1.41 (m, 4H), 1.41-1.26 (m, 9H), 1.22 (dd, J=7.1, 4.0 Hz, 3H); 13C NMR (101 MHz, DMSO-D6) δ 174.32 (s), 161.80 (s), 160.32 (d, J=2.6 Hz), 153.82 (d, J=3.4 Hz), 147.61 (s), 147.16 (d, J=3.5 Hz), 143.63 (s), 133.00 (s), 122.72 (s), 108.72 (s), 108.19 (s), 101.41 (d, J=6.3 Hz), 78.47 (s), 60.38 (s), 47.49 (s), 47.00 (s), 45.10 (d, J=4.8 Hz), 28.50 (s), 28.49-27.14 (m), 22.57 (s), 14.09 (s). HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 576.6540, found 577.9603.
14e (30 mg, 43%). 1H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J=5.1 Hz, 1H), 7.26 (s, 1H), 7.04 (d, J=8.0 Hz, 1H), 6.74 (d, J=8.2 Hz, 1H), 6.10 (d, J=25.8 Hz, 1H), 4.55 (t, J=8.7 Hz, 2H), 4.17-4.09 (m, 2H), 3.78 (d, J=5.6 Hz, 3H), 3.20 (s, 1H), 3.16 (t, J=8.6 Hz, 2H), 3.07 (d, J=5.4 Hz, 1H), 3.02-2.76 (m, 4H), 1.73 (d, J=26.4 Hz, 2H), 1.37 (dd, J=13.5, 7.6 Hz, 9H), 1.24-1.20 (m, 5H), 1.13 (dd, J=14.9, 7.7 Hz, 1H), 0.87-0.78 (m, 1H). HRMS (ESI+) calculated for C31H39N6O5 [M+H]+: 575.2976, found 575.2137.
15a (64 mg, 69%). 1H NMR (400 MHz, DMSO-d6) δ 8.35 (d, J=4.9 Hz, 1H), 7.78-7.62 (m, 2H), 7.58-7.43 (m, 2H), 6.41 (d, J=48.8 Hz, 1H), 4.17-4.09 (m, 2H), 3.85-3.79 (m, 1H), 3.72-3.53 (m, 2H), 3.12 (dd, J=32.3, 25.7 Hz, 2H), 3.05-2.93 (m, 2H), 2.93-2.81 (m, 2H), 1.69-1.56 (m, 2H), 1.40 (dt, J=25.1, 11.5 Hz, 11H), 1.24-1.20 (m, 4H). HRMS (ESI+) calculated for C29H33F4N6O4 [M+H]+: 605.2494, found 605.2074.
15b (98 mg, 98%)1H NMR (400 MHz, DMSO) δ 8.35 (d, J=5.2 Hz, 1H), 7.61 (d, J=8.4 Hz, 2H), 7.54 (s, 1H), 7.33-7.27 (m, 1H), 6.45 (s, 1H), 4.18-4.11 (m, 2H), 3.88-3.80 (m, 1H), 3.76-3.54 (m, 2H), 3.28-2.81 (m, 7H), 1.88-1.49 (m, 4H), 1.39 (d, J=5.7 Hz, 9H), 1.24 (dd, J=5.5, 1.4 Hz, 3H). MS (ESI+) calculated for C29H35Cl2N6O4+ [M+H]+: 601.2091, Found 601.0306.
15c (29 mg, 61%). 1H NMR (400 MHz, DMSO-D6) δ 8.22 (d, J=5.2 Hz, 1H), 8.00 (s, 1H), 7.93-7.86 (m, 3H), 7.53 (p, J=6.8 Hz, 2H), 7.44 (s, 2H), 6.25 (s, 1H), 4.14 (dt, J=7.1, 4.6 Hz, 2H), 3.84 (dd, J=8.8, 5.9 Hz, 1H), 3.67 (dd, J=29.5, 25.0 Hz, 1H), 3.56-3.35 (m, 2H), 3.21 (d, J=27.7 Hz, 4H), 3.01 (dt, J=12.7, 7.7 Hz, 1H), 2.94-2.86 (m, 1H), 2.84-2.53 (m, 1H), 1.95-1.47 (m, 2H), 1.46-1.28 (m, 9H), 1.23 (t, J=3.5 Hz, 3H), 1.03-0.74 (m, 1H). 13C NMR (101 MHz, DMSO-D6) δ 174.31 (s), 161.78 (s), 160.46 (d, J=4.6 Hz), 153.75 (d, J=5.8 Hz), 144.23 (s), 132.59 (d, J=2.4 Hz), 128.18 (s), 127.57 (s), 126.74 (s), 126.63-126.57 (m), 126.33 (d, J=40.9 Hz), 125.39 (s), 123.95 (s), 102.31 (s), 78.45 (s), 60.41 (s), 48.60 (s), 47.51 (s), 46.81 (s), 45.18 (s), 29.08-28.72 (m), 28.72-28.59 (m), 28.72-27.65 (m), 14.11 (s); HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 582.7050, found 583.9995.
15d (99 mg, 92%). 1H NMR (400 MHz, DMSO-D6) δ 8.25 (d, J=5.2 Hz, 1H), 7.55-7.34 (m, 1H), 6.91 (d, J=8.0 Hz, 2H), 6.81 (dd, J=8.1, 1.6 Hz, 1H), 6.23-6.11 (m, 1H), 6.05 (s, 2H), 4.13 (dt, J=7.0, 5.2 Hz, 2H), 3.86-3.74 (m, 2H), 3.66 (d, J=2.9 Hz, 1H), 3.62-3.34 (m, 2H), 3.28-3.01 (m, 3H), 2.89 (ddd, J=20.3, 14.9, 7.5 Hz, 3H), 2.62 (dd, J=27.7, 19.4 Hz, 1H), 1.75 (d, J=13.1 Hz, 2H), 1.37 (dt, J=14.5, 10.6 Hz, 9H), 1.21 (dd, J=7.1, 3.1 Hz, 3H); 13C NMR (101 MHz, DMSO-D6) δ 174.32 (s), 161.81 (s), 160.31 (d, J=8.1 Hz), 153.86 (s), 153.62 (s), 147.61 (s), 147.18 (d, J=3.3 Hz), 143.62 (s), 137.61 (s), 122.72 (s), 108.73 (s), 108.20 (s), 101.36 (d, J=5.1 Hz), 78.50 (s), 60.39 (s), 47.51 (s), 46.96 (s), 45.11 (d, J=5.3 Hz), 28.33 (d, J=33.9 Hz), 27.88 (d, J=13.9 Hz), 23.55 (s), 14.10 (s); HRMS (ESI+) calculated for C32H36N6O4 [M+H]+: 576.6540, found 577.9503.
15e (28 mg, 40%). 1H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J=5.1 Hz, 1H), 7.26 (s, 1H), 7.04 (d, J=8.0 Hz, 1H), 6.74 (d, J=8.2 Hz, 1H), 6.13 (s, 1H), 4.56 (t, J=8.7 Hz, 2H), 4.18-4.08 (m, 2H), 3.78 (d, J=5.5 Hz, 3H), 3.14 (dd, J=24.8, 16.1 Hz, 4H), 2.88 (ddd, J=20.2, 14.7, 7.2 Hz, 4H), 1.74 (d, J=25.4 Hz, 2H), 1.38 (d, J=3.5 Hz, 9H), 1.25-1.19 (m, 5H), 1.18-1.06 (m, 1H), 0.88-0.82 (m, 1H); HRMS (ESI+) calculated for C31H39N6O5 [M+H]+: 575.2976, found 575.5378.
Compound 12a (25 mg, 0.043 mmol) was dissolved in MC (1.072 mL), 20% TFA (v/v) (268 μL) was added, and then the mixture was stirred for 30 minutes. After confirming the termination of the reaction, the resulting product was concentrated with MC three times. The resulting concentrate was dissolved in THF (1.15 mL, 0.1M), TEA (40 μL, 0.230 mmol) was added, and cyclopropanecarbonyl chloride (10.43 μL, 0.1.15 mmol) was slowly added and stirred. After confirming the termination of the reaction, the resulting product was extracted with ethyl acetate (EtOAc) and washed with water and brine. After drying water with anhydrous magnesium sulfate (MgSO4), filtration and concentration were performed. Purification and separation were performed through column chromatography, thereby obtaining Compound 16a (44 mg, 66.8%).
16a (44 mg, 66.8%). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (s, 1H), 7.80 (t, J=37.9 Hz, 3H), 7.52 (s, 1H), 6.51 (d, J=17.9 Hz, 1H), 4.13 (d, J=6.3 Hz, 2H), 3.84 (s, 1H), 3.73 (s, 1H), 3.50 (d, J=52.3 Hz, 2H), 3.19 (d, J=24.7 Hz, 3H), 3.05-2.83 (m, 2H), 1.75 (d, J=29.3 Hz, 1H), 1.53 (s, 1H), 1.22 (s, 4H), 0.83 (d, J=9.4 Hz, 1H), 0.71 (s, 4H); HRMS (ESI+) calculated for C28H29F4N6O3 [M+H]+: 573.2232, found 573.2693.
16b (22 mg, 58%)1H NMR (400 MHz, DMSO) δ 8.37 (dd, J=13.9, 5.2 Hz, 1H), 7.92-7.77 (m, 1H), 7.66 (s, 1H), 7.63 (dd, J=8.4, 2.3 Hz, 1H), 7.32 (t, J=6.1 Hz, 1H), 6.50 (d, J=33.9 Hz, 1H), 4.13 (dt, J=10.4, 4.3 Hz, 2H), 3.83 (d, J=8.6 Hz, 1H), 3.75 (d, J=7.1 Hz, 1H), 3.58 (s, 1H), 3.46 (s, 1H), 3.23 (d, J=9.3 Hz, 2H), 3.15 (dd, J=16.2, 9.5 Hz, 2H), 2.98 (dd, J=15.5, 8.5 Hz, 1H), 2.87 (dd, J=14.8, 5.3 Hz, 1H), 1.87 (d, J=51.0 Hz, 2H), 1.76-1.67 (m, 1H), 1.24-1.19 (m, 3H), 0.71 (dd, J=8.2, 4.2 Hz, 4H); MS(ESI+) calculated for C27H29C12N6O3+ [M+H]+: 555.1673, found 555.9500.
16c (13.9 mg, 55%). 1H NMR (400 MHz, DMSO-D6) δ 8.36-8.23 (m, 1H), 8.00 (d, J=5.8 Hz, 1H), 7.90 (dd, J=10.3, 7.1 Hz, 3H), 7.74 (dd, J=26.5, 6.3 Hz, 1H), 7.58-7.41 (m, 3H), 6.31 (dd, J=72.1, 29.8 Hz, 1H), 4.20-4.11 (m, 2H), 3.86 (t, J=11.6 Hz, 1H), 3.64 (d, J=70.7 Hz, 1H), 3.53-3.33 (m, 2H), 3.30-3.04 (m, 4H), 2.95 (ddd, J=19.5, 14.1, 5.1 Hz, 2H), 1.97-1.34 (m, 3H), 1.23 (td, J=7.1, 1.9 Hz, 3H), 0.74-0.57 (m, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.33 (s), 170.32 (s), 162.17-161.71 (m), 157.04 (s), 144.31 (s), 132.53 (t, J=6.5 Hz), 131.26 (s), 128.13 (s), 127.47 (d, J=12.8 Hz), 126.64 (d, J=17.3 Hz), 126.08 (s), 122.93 (s), 102.19 (s), 60.44 (s), 51.95 (s), 51.12 (s), 49.06 (s), 45.20 (s), 30.80 (s), 28.16 (s), 20.75 (s), 14.11 (s), 11.57 (s), 6.91 (d, J=5.3 Hz). HRMS (ESI+) calculated for C31H32N6O3 [M+H]+: 536.6360, found 537.9465.
16d (42.4 mg, 26%). 1H NMR (400 MHz, DMSO-D6) δ 8.29 (dd, J=9.9, 5.3 Hz, 1H), 7.81 (d, J=24.7 Hz, 1H), 6.98-6.87 (m, 2H), 6.83 (dd, J=8.1, 1.6 Hz, 1H), 6.31 (d, J=23.4 Hz, 1H), 6.09-6.00 (m, 2H), 4.13 (q, J=7.1 Hz, 2H), 4.06-3.75 (m, 2H), 3.74-3.55 (m, 1H), 3.54-3.34 (m, 2H), 3.30-3.03 (m, 3H), 2.90 (ddd, J=20.4, 14.8, 7.5 Hz, 2H), 2.14-1.57 (m, 3H), 1.25-1.17 (m, 3H), 0.71 (dd, J=9.3, 5.7 Hz, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.35 (s), 170.92 (s), 162.46 (s), 162.03 (d, J=13.6 Hz), 147.65 (s), 147.14 (d, J=5.3 Hz), 143.70 (s), 132.98 (s), 127.09 (s), 122.68 (s), 108.69 (s), 108.18 (s), 101.44 (d, J=8.5 Hz), 60.40 (s), 54.90 (s), 50.74 (s), 45.11 (s), 44.42 (s), 30.95 (s), 28.50 (s), 28.16 (s), 14.09 (s), 11.96 (s), 6.92 (d, J=4.3 Hz); HIRMS (ESI+) calculated for C28H30N6O5 [M+H]+: 530.5850, found 531.8253.
16e (9.1 mg, 33%). 1H NMR (400 MHz, DMSO-d6) δ 8.27 (dd, J=11.8, 5.3 Hz, 1H), 7.79 (d, J=22.4 Hz, 1H), 7.29 (s, 1H), 7.05 (d, J=8.2 Hz, 1H), 6.77-6.72 (m, 1H), 6.25 (d, J=34.8 Hz, 1H), 4.54 (dd, J=8.5, 6.1 Hz, 2H), 4.15-4.10 (m, 2H), 3.80 (s, 2H), 3.46 (d, J=5.4 Hz, 1H), 3.28-3.08 (m, 6H), 2.99-2.90 (m, 1H), 2.84 (dd, J=14.5, 5.5 Hz, 1H), 1.99-1.90 (m, 1H), 1.75 (dt, J=12.4, 6.2 Hz, 1H), 1.48 (ddd, J=12.6, 5.9, 3.8 Hz, 1H), 1.21 (dd, J=7.0, 1.3 Hz, 3H), 0.81 (t, J=2.7 Hz, 1H), 0.77 (dd, J=4.9, 2.1 Hz, 2H), 0.71 (d, J=2.0 Hz, 2H); HIRMS (ESI+) calculated for C29H33N6O4 [M+H]+: 529.2558, found 529.3768.
17a (47.3 mg, 70%). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J=10.2, 5.2 Hz, 1H), 7.94-7.68 (m, 3H), 7.57-7.48 (m, 1H), 6.51 (d, J=19.8 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 3.85 (s, 1H), 3.73 (s, 1H), 3.56 (s, 1H), 3.43 (d, J=7.2 Hz, 1H), 3.27-3.10 (m, 3H), 3.04-2.94 (m, 1H), 2.88 (dd, J=15.5, 4.8 Hz, 1H), 1.72 (s, 1H), 1.64-1.44 (m, 1H), 1.30-1.16 (m, 4H), 0.70 (dd, J=9.4, 5.4 Hz, 5H); IRMS (ESI+) calculated for C28H29F4N6O4 [M+H]+: 573.2232, found 573.2333.
17b (27 mg, 78%)1H NMR (400 MHz, DMSO) δ 8.37 (dd, J=14.1, 5.2 Hz, 1H), 7.93-7.79 (m, 1H), 7.66 (s, 1H), 7.63 (dd, J=8.4, 2.3 Hz, 1H), 7.35-7.29 (m, 1H), 6.50 (d, J=34.8 Hz, 1H), 4.13 (q, J=7.1 Hz, 2H), 3.84 (s, 1H), 3.74 (s, 1H), 3.66-3.40 (m, 2H), 3.29-2.83 (m, 6H), 2.01-1.91 (m, 1H), 1.75-1.70 (m, 1H), 1.55 (s, 1H), 1.22 (dd, J=7.1, 5.0 Hz, 3H), 0.71 (dd, J=8.2, 4.2 Hz, 4H); MS(ESI+) calculated for C27H29Cl2N6O3 [M+H]+: 555.1673, Found 555.9140.
17c (13.8 mg, 76%). 1H NMR (400 MHz, DMSO-D6) δ 8.34-8.23 (m, 1H), 8.00 (d, J=6.1 Hz, 1H), 7.90 (m, 3H), 7.75 (dd, J=26.5, 6.3 Hz, 1H), 7.57-7.49 (m, 2H), 7.45 (t, J=8.1 Hz, 1H), 6.40 (d, J=51.6 Hz, 1H), 4.19-4.11 (m, 2H), 3.87 (m, 1H), 3.57 (t, J=64.8 Hz, 2H), 3.28 (dd, J=18.4, 11.6 Hz, 4H), 3.22-3.14 (m, 1H), 3.13-2.96 (m, 2H), 2.91 (dd, J=14.6, 5.4 Hz, 1H), 1.60 (s, 2H), 1.23 (tt, J=7.0, 1.5 Hz, 3H), 0.70 (dd, J=20.0, 15.6 Hz, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.33 (s), 170.32 (s), 162.07-161.71 (m), 156.86 (s), 144.32 (s), 132.73-132.24 (m), 128.84 (s), 128.17 (d, J=8.4 Hz), 127.95-126.09 (m), 126.64 (d, J=16.4 Hz), 126.45 (t, J=32.4 Hz), 101.83 (s), 59.74 (s), 51.07 (s), 50.59 (s), 45.20 (s), 43.59 (s), 30.80 (s), 29.02 (s), 28.15 (s), 20.74 (s), 14.07 (s), 7.63 (s), 6.91 (s); HRMS (ESI+) calculated for C31H32N6O3 [M+H]+: 536.6360, found 537.6224.
17d (61.8 mg, 26%). 1H NMR (400 MHz, DMSO-D6) δ 8.29 (dd, J=9.9, 5.3 Hz, 1H), 7.82 (d, J=25.5 Hz, 1H), 6.96-6.88 (m, 2H), 6.84 (dd, J=8.1, 1.6 Hz, 1H), 6.30 (d, J=22.7 Hz, 1H), 6.09-5.99 (m, 2H), 4.13 (q, J=7.1 Hz, 2H), 4.08-3.73 (m, 2H), 3.73-3.54 (m, 1H), 3.53-3.35 (m, 2H), 3.32-3.02 (m, 3H), 2.90 (ddd, J=20.5, 14.8, 7.5 Hz, 2H), 1.82 (dddd, J=53.0, 47.8, 42.4, 24.2 Hz, 3H), 1.25-1.17 (m, 3H), 0.75-0.66 (m, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.35 (s), 170.93 (s), 162.64 (s), 162.02 (d, J=10.4 Hz), 156.81 (s), 147.65 (s), 147.17 (s), 143.73 (s), 132.96 (s), 125.86 (s), 122.69 (s), 108.70 (s), 108.18 (s), 101.44 (d, J=8.5 Hz), 60.40 (s), 54.90 (s), 51.30 (s), 45.12 (s), 44.42 (s), 30.96 (s), 28.89 (d, J=77.5 Hz), 28.17 (s), 14.08 (s), 11.97 (s), 6.92 (d, J=4.5 Hz); HRMS (ESI+) calculated for C28H30N6O5 [M+H]+: 530.5850, found 531.1412.
17e (15.6 mg, 41%). 1H NMR (400 MHz, DMSO-d6) δ 8.27 (dd, J=12.0, 5.2 Hz, 1H), 7.87-7.71 (m, 1H), 7.29 (s, 1H), 7.05 (d, J=8.2 Hz, 1H), 6.74 (dd, J=8.3, 1.8 Hz, 1H), 6.35-6.06 (m, 1H), 4.59-4.50 (m, 2H), 4.13 (dd, J=14.0, 7.0 Hz, 2H), 3.76 (d, J=26.0 Hz, 2H), 3.46 (d, J=5.4 Hz, 1H), 3.30-3.09 (m, 6H), 3.00-2.90 (m, 1H), 2.84 (dd, J=14.5, 5.1 Hz, 1H), 1.93 (dd, J=18.0, 8.6 Hz, 1H), 1.75 (dt, J=12.8, 6.3 Hz, 1H), 1.49 (tt, J=7.7, 4.8 Hz, 1H), 1.21 (t, J=7.1 Hz, 3H), 0.81 (t, J=2.6 Hz, 1H), 0.79-0.76 (m, 2H), 0.73-0.71 (m, 2H); HRMS (ESI+) calculated for C29H33N6O4 [M+H]+: 529.2558, found 529.2688.
18a (54.6 mg, 75%)1H NMR (400 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.79-7.57 (m, 3H), 7.49 (s, 1H), 6.43 (d, J=39.3 Hz, 1H), 4.16-4.10 (m, 2H), 4.00 (d, J=35.6 Hz, 2H), 3.83 (s, 1H), 3.16 (d, J=35.7 Hz, 2H), 3.07-2.82 (m, 4H), 1.93 (s, 1H), 1.66 (s, 2H), 1.49 (ddd, J=12.4, 7.7, 4.9 Hz, 2H), 1.24-1.20 (m, 3H), 0.59 (d, J=81.9 Hz, 5H); HRMS (ESI+) calculated for C29H31F4N6O3 [M+H]+: 587.2388, found 587.2401.
18b (29 mg, 77%)1H NMR (400 MHz, DMSO) δ 8.35 (s, 1H), 7.60 (d, J=8.0 Hz, 3H), 7.28 (s, 1H), 6.47 (s, 1H), 4.18-4.08 (m, 2H), 3.94 (s, 1H), 3.83 (d, J=5.4 Hz, 2H), 3.28-2.55 (m, 7H), 1.78 (dd, J=11.9, 6.1 Hz, 1H), 1.72-1.59 (m, 2H), 1.41 (ddd, J=20.2, 10.4, 3.9 Hz, 2H), 1.22 (s, 3H), 0.73-0.44 (m, 4H); MS(ESI+) calculated for C28H31Cl2N6O3+ [M+H]+: 569.1829 Found 569.8487.
18c (25 mg, 40%). 1H NMR (400 MHz, MeOD) δ 8.20 (d, J=5.2 Hz, 1H), 8.00 (s, 1H), 7.90 (dd, J=12.6, 5.6 Hz, 3H), 7.59-7.52 (m, 2H), 7.44 (d, J=8.3 Hz, 1H), 6.35 (s, 1H), 4.28-4.20 (m, 2H), 3.93-3.87 (m, 1H), 3.54 (dd, J=80.1, 58.5 Hz, 3H), 3.28 (s, 1H), 3.07 (ddd, J=31.5, 20.3, 11.9 Hz, 4H), 1.52-1.35 (m, 9H), 1.34-1.30 (m, 6H), 0.91 (d, J=7.0 Hz, 2H); 13C NMR (101 MHz, DMSO-D6) δ 174.34 (s), 170.84 (s), 161.73 (s), 160.59 (d, J=4.9 Hz), 149.70 (s), 144.25 (s), 132.56 (t, J=4.8 Hz), 129.60 (s), 128.67 (s), 128.21 (d, J=6.4 Hz), 127.55 (s), 126.62 (d, J=19.6 Hz), 125.99 (s), 102.11 (s), 60.42 (s), 45.18 (s), 41.78 (s), 29.77 (s), 28.27 (d, J=19.3 Hz), 22.65 (s), 14.12 (s), 10.44 (s), 6.72 (d, J=30.3 Hz), 6.55-6.46 (m); HRMS (ESI+) calculated for C32H34N6O3 [M+H]+: 550.6630, found 551.7372.
18d (8.1 mg, 26%). 1H NMR (400 MHz, MeOD) δ 8.24 (d, J=24.3 Hz, 1H), 6.92-6.84 (m, 3H), 6.26 (s, 1H), 6.01 (s, 2H), 4.26-4.18 (m, 2H), 4.04 (d, J=58.6 Hz, 2H), 3.85 (s, 1H), 3.21 (d, J=44.6 Hz, 2H), 3.08-2.95 (m, 2H), 2.10-1.94 (m, 2H), 1.80 (s, 2H), 1.54 (d, J=57.0 Hz, 3H), 1.34-1.29 (m, 6H), 0.92 (t, J=6.8 Hz, 2H), 0.87-0.74 (m, 2H). 13C NMR (101 MHz, DMSO-D6) δ 174.36 (s), 170.95 (s), 162.73 (s), 161.79 (s), 160.46 (s), 156.90 (s), 147.57 (s), 147.13 (s), 143.64 (s), 133.02 (s), 122.65 (s), 108.98-108.73 (m), 108.40 (d, J=45.5 Hz), 101.39 (d, J=2.5 Hz), 60.39 (s), 46.23 (s), 45.09 (s), 30.68 (s), 28.43 (s), 28.16 (s), 22.93 (s), 14.11 (s), 10.42 (s), 6.83 (d, J=3.6 Hz). HRMS (ESI+) calculated for C29H32N6O5 [M+H]+: 544.6120, found 545.9041.
18e (10.7 mg, 38%)1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.50 (d, J=28.0 Hz, 1H), 7.26 (s, 1H), 7.03 (s, 1H), 6.73 (d, J=8.1 Hz, 1H), 6.15 (d, J=48.8 Hz, 1H), 4.55 (t, J=6.9 Hz, 2H), 4.13 (dd, J=7.1, 3.2 Hz, 2H), 3.79 (s, 1H), 3.17 (dd, J=16.0, 8.3 Hz, 6H), 2.94 (dd, J=17.9, 11.8 Hz, 2H), 2.83 (dd, J=16.8, 6.0 Hz, 2H), 1.96 (d, J=12.3 Hz, 1H), 1.78 (s, 3H), 1.24-1.19 (m, 4H), 0.71 (s, 3H), 0.60 (s, 1H), 0.49 (s, 1H); HIRMS (ESI+) calculated for C30H35N6O4 [M+H]+: 543.2714, found 543.3116.
19a (40 mg, 66%) 1H NMR (400 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.78-7.57 (m, 3H), 7.50 (s, 1H), 6.44 (d, J=39.0 Hz, 1H), 4.16-4.10 (m, 2H), 4.01 (d, J=34.9 Hz, 2H), 3.83 (s, 1H), 3.15 (d, J=38.2 Hz, 2H), 3.05-2.82 (m, 4H), 1.91 (s, 1H), 1.77 (dd, J=13.0, 7.2 Hz, 1H), 1.65 (s, 3H), 1.22 (d, J=4.8 Hz, 3H), 0.59 (d, J=81.6 Hz, 5H); HIRMS (ESI+) calculated for C29H31F4N6O3 [M+H]+: 587.2388, found 587.2401.
19b (50 mg, 74%)1H NMR (400 MHz, DMSO) δ 8.37 (s, 1H), 7.72-7.50 (m, 3H), 7.30 (s, 1H), 6.43 (d, J=47.6 Hz, 1H), 4.20-4.10 (m, 2H), 3.97 (d, J=6.3 Hz, 1H), 3.90-3.47 (m, 2H), 3.32-2.59 (m, 7H), 1.95 (s, 1H), 1.64 (dd, J=15.2, 11.2 Hz, 2H), 1.51-1.34 (m, 2H), 1.24 (d, J=5.6 Hz, 3H), 0.79-0.44 (m, 4H); MS(ESI+) calculated for C28H31Cl2N6O3+ [M+H]+: 569.1829, found 569.9567.
19c (12.1 mg, 57%). 1H NMR (400 MHz, DMSO-D6) δ 8.24 (s, 1H), 7.91 (dd, J=26.0, 19.5 Hz, 4H), 7.58-7.32 (m, 4H), 6.23 (d, J=66.0 Hz, 1H), 4.18-4.11 (m, 2H), 3.84 (s, 2H), 3.54-3.34 (m, 2H), 3.30-2.80 (m, 6H), 2.03-1.84 (m, 1H), 1.82-1.35 (m, 3H), 1.23 (td, J=7.1, 4.0 Hz, 3H), 0.93-0.77 (m, 1H), 0.63 (d, J=57.5 Hz, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.33 (s), 161.94-161.66 (m), 156.84 (s), 144.24 (s), 132.96-132.34 (m), 129.28 (s), 128.91 (s), 128.20 (d, J=4.4 Hz), 127.94-126.48 (m), 126.48-126.12 (m), 121.04 (s), 101.23 (s), 60.43 (s), 54.90 (s), 47.72 (s), 45.18 (s), 44.46 (s), 28.27 (d, J=19.1 Hz), 22.62 (s), 14.12 (s), 10.43 (s), 6.83 (d, J=7.9 Hz); IRMS (ESI+) calculated for C32H34N6O3 [M+H]+: 550.6630, found 551.8812.
19d (7.6 mg, 29%). 1H NMR (400 MHz, DMSO-D6) δ 8.32-8.18 (m, 1H), 7.53 (d, J=39.0 Hz, 1H), 6.86 (dd, J=34.3, 7.9 Hz, 3H), 6.16 (t, J=25.4 Hz, 1H), 6.05 (s, 2H), 4.32-3.68 (m, 5H), 3.33 (s, 2H), 3.18-2.77 (m, 5H), 1.92 (d, J=26.5 Hz, 1H), 1.76 (d, J=23.6 Hz, 2H), 1.34 (dd, J=60.3, 29.5 Hz, 2H), 1.22 (t, J=5.8 Hz, 3H), 0.82-0.37 (m, 4H); 13C NMR (101 MHz, DMSO-D6) δ 174.36 (s), 170.94 (s), 161.79 (s), 160.43 (s), 149.35 (s), 147.57 (s), 147.13 (s), 143.63 (s), 132.99 (s), 129.34 (s), 122.65 (s), 108.65 (s), 108.18 (s), 101.34 (d, J=5.9 Hz), 60.39 (s), 49.10 (s), 46.79 (s), 45.08 (s), 41.91 (s), 28.43 (s), 28.16 (s), 26.76 (s), 23.00 (s), 14.11 (s), 10.42 (s), 6.72 (d, J=25.3 Hz); HRMS (ESI+) calculated for C29H32N6O5 [M+H]+: 544.6120, found 545.9761.
19e (11 mg, 41.7%)1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.51 (d, J=34.7 Hz, 1H), 7.26 (s, 1H), 7.03 (s, 1H), 6.74 (t, J=9.9 Hz, 1H), 6.15 (d, J=50.0 Hz, 1H), 4.60-4.49 (m, 2H), 4.15-4.09 (m, 2H), 3.79 (s, 1H), 3.28-2.99 (m, 7H), 3.00-2.77 (m, 3H), 1.94 (dd, J=13.1, 6.1 Hz, 1H), 1.75 (d, J=25.4 Hz, 3H), 1.27-1.18 (m, 4H), 0.71 (s, 3H), 0.60 (s, 1H), 0.49 (s, 1H); HRMS (ESI+) calculated for C30H35N6O4 [M+H]+: 543.2714, found 543.2036.
Compound 16a (23.3 mg, 0.040 mmol), which had been obtained in the previous experiment, was diluted in ammonia in 7N-methanol (0.91 mL) and then stirred at room temperature for 3 days. It was able to be confirmed that the initial ethyl ester develops into methyl ester, and an amide is formed from the methyl ester. After confirming the completion of the reactor, the resulting product was extracted with ethyl acetate (EtOAc), and the organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using MC and MeOH (MC:MeOH=20:1) as mobile phase, thereby obtaining Compound 20a (13.8 mg, 81.9%).
20a (13.8 mg, 81.9%). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J=9.4, 5.2 Hz, 1H), 7.92-7.66 (m, 3H), 7.59-7.40 (m, 2H), 6.94 (s, 1H), 6.51 (d, J=25.2 Hz, 1H), 3.72 (s, 1H), 3.69-3.50 (m, 2H), 3.40 (dd, J=17.8, 7.3 Hz, 1H), 3.29-3.21 (m, 1H), 3.08 (d, J=7.1 Hz, 2H), 2.95-2.84 (m, 1H), 2.77 (dd, J=14.6, 5.6 Hz, 1H), 1.83-1.65 (m, 2H), 1.61-1.43 (m, 1H), 0.85 (ddd, J=8.6, 5.2, 2.1 Hz, 1H), 0.76-0.64 (m, 4H); HRMS (ESI+) calculated for C26H26F4N7O2 [M+H]+: 544.2079, found 544.2117.
20b (4 mg, 49%) 1H NMR (400 MHz, DMSO) δ 8.38 (dd, J=13.5, 5.2 Hz, 1H), 7.83 (d, J=24.1 Hz, 1H), 7.66 (s, 1H), 7.63 (dd, J=8.4, 2.1 Hz, 1H), 7.45 (s, 1H), 7.31 (t, J=6.8 Hz, 1H), 6.94 (s, 1H), 6.50 (d, J=38.8 Hz, 1H), 3.77 (d, J=29.0 Hz, 1H), 3.66-3.59 (m, 1H), 3.55-3.44 (m, 2H), 3.40 (ddd, J=10.0, 6.9, 4.3 Hz, 2H), 3.07 (s, 2H), 2.93-2.84 (m, 1H), 2.76 (dd, J=14.1, 4.9 Hz, 1H), 1.83-1.67 (m, 2H), 1.50-1.40 (m, 1H), 0.76-0.65 (m, 4H); MS(ESI+) calculated for C25H26Cl2N7O2+ [M+H]+: 526.1520, found 526.7123.
20c (8.4 mg, 68%). 1H NMR (400 MHz, DMSO-D6) δ 8.34-8.24 (m, 1H), 7.99 (d, J=6.1 Hz, 1H), 7.90 (t, J=8.7 Hz, 3H), 7.72 (dd, J=25.4, 6.1 Hz, 1H), 7.59-7.35 (m, 4H), 6.94 (s, 1H), 6.40 (d, J=58.9 Hz, 1H), 3.65 (s, 2H), 3.58-3.35 (m, 2H), 3.16 (dd, J=28.0, 14.7 Hz, 4H), 2.97-2.75 (m, 2H), 1.65 (d, J=54.6 Hz, 2H), 1.23 (s, 1H), 0.68 (d, J=22.3 Hz, 4H); HRMS (ESI+) calculated for C29H29N7O2 [M+H]+: 507.5980, found 508.5647.
20d (26.7 mg, 92%). 1H NMR (400 MHz, DMSO-D6) δ 8.34-8.25 (m, 1H), 7.92 (d, J=4.6 Hz, 1H), 7.79 (d, J=24.3 Hz, 1H), 6.98-6.87 (m, 2H), 6.83 (dd, J=8.1, 1.4 Hz, 1H), 6.40-6.15 (m, 1H), 6.09-5.98 (m, 2H), 3.82-3.66 (m, 1H), 3.64-3.54 (m, 1H), 3.39 (d, J=40.4 Hz, 3H), 3.32-2.94 (m, 3H), 2.92-2.66 (m, 2H), 2.65-2.58 (m, 3H), 1.98 (d, J=3.5 Hz, 2H), 1.74-1.39 (m, 1H), 0.70 (dd, J=10.7, 7.4 Hz, 4H). HRMS (ESI+) calculated for C26H27N7O4 [M+H]+: 501.5470, found 502.5154.
20e (4.1 mg, 54%). 1H NMR (400 MHz, DMSO-d6) δ 8.30-8.23 (m, 1H), 7.75 (d, J=24.3 Hz, 1H), 7.44 (s, 1H), 7.28 (s, 1H), 7.04 (d, J=8.3 Hz, 1H), 6.90 (s, 1H), 6.73 (dd, J=8.3, 2.4 Hz, 1H), 6.25 (d, J=41.7 Hz, 1H), 4.54 (dd, J=15.4, 8.2 Hz, 2H), 3.99 (s, 1H), 3.77 (d, J=8.5 Hz, 1H), 3.59 (s, 2H), 3.49 (t, J=4.2 Hz, 1H), 3.15 (t, J=7.3 Hz, 2H), 3.10-2.98 (m, 2H), 2.91-2.81 (m, 1H), 2.76-2.66 (m, 1H), 1.98-1.87 (m, 1H), 1.76 (dd, J=12.2, 5.9 Hz, 1H), 1.57 (d, J=24.5 Hz, 1H), 0.69 (t, J=19.8 Hz, 5H); HRMS (ESI+) calculated for C27H30N7O3 [M+H]+: 500.2405, found 500.2469.
21a (15 mg, 78.8%). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J=9.4, 5.2 Hz, 1H), 7.93-7.67 (m, 3H), 7.51 (dt, J=14.0, 5.5 Hz, 1H), 7.45 (s, 1H), 6.94 (s, 1H), 6.52 (d, J=25.5 Hz, 1H), 3.72 (s, 1H), 3.68-3.52 (m, 2H), 3.40 (dd, J=18.8, 9.9 Hz, 1H), 3.28 (s, 1H), 3.08 (d, J=7.1 Hz, 2H), 2.95-2.85 (m, 1H), 2.77 (dd, J=14.5, 5.5 Hz, 1H), 1.82-1.63 (m, 2H), 1.60-1.44 (m, 1H), 0.88-0.83 (m, 1H), 0.72-0.66 (m, 4H); HRMS (ESI+) calculated for C26H26F4N7O2 [M+H]+: 544.2079, found 544.2117.
21b (17 mg, 80%)1H NMR (400 MHz, DMSO) δ 8.37 (dd, J=13.4, 5.2 Hz, 1H), 7.83 (ddd, J=15.2, 8.2, 3.6 Hz, 1H), 7.68-7.60 (m, 2H), 7.45 (s, 1H), 7.31 (t, J=7.0 Hz, 1H), 6.94 (s, 1H), 6.50 (d, J=38.1 Hz, 1H), 3.74 (s, 1H), 3.62 (dd, J=14.7, 7.0 Hz, 2H), 3.51-3.41 (m, 1H), 3.27-2.70 (m, 6H), 1.74 (dd, J=17.6, 11.6 Hz, 2H), 1.55 (s, 1H), 0.76-0.63 (m, 4H); MS(ESI+) calculated for C25H26Cl2N7O2 [M+H]+: 526.1520, found 526.8563.
21c (8 mg, 66%). 1H NMR (400 MHz, DMSO-D6) δ 8.35-8.23 (m, 1H), 7.99 (d, J=6.2 Hz, 1H), 7.90 (t, J=8.8 Hz, 3H), 7.72 (dd, J=25.3, 6.3 Hz, 1H), 7.58-7.40 (m, 4H), 6.94 (s, 1H), 6.41 (d, J=54.2 Hz, 1H), 3.84-3.59 (m, 2H), 3.48 (d, J=45.9 Hz, 1H), 3.03 (dd, J=72.8, 9.5 Hz, 4H), 2.93-2.75 (m, 2H), 1.99 (s, 1H), 1.58 (s, 2H), 1.39-1.19 (m, 1H), 0.76-0.55 (m, 4H); 13C NMR (101 MHz, DMSO-D6) δ 175.87 (s), 170.33 (s), 162.17-161.47 (m), 156.91 (s), 144.80 (s), 132.84-132.03 (m), 129.74 (s), 128.66-128.41 (m), 127.57 (dd, J=47.3, 36.9 Hz), 126.60 (d, J=13.0 Hz), 102.13 (s), 59.74 (s), 51.13 (s), 46.12 (s), 43.59 (s), 30.83 (s), 28.91 (s), 28.68 (s), 14.08 (s), 6.93 (d, J=2.8 Hz); HRMS (ESI+) calculated for C29H29N7O2 [M+H]+: 507.5980, found 508.5647.
21d (18.3 mg, 65%). 1H NMR (400 MHz, DMSO-D6) δ 8.36-8.25 (m, 1H), 7.79 (d, J=24.6 Hz, 1H), 7.44 (s, 1H), 6.98-6.87 (m, 3H), 6.82 (dd, J=8.1, 1.3 Hz, 1H), 6.31 (d, J=28.0 Hz, 1H), 6.09-5.98 (m, 2H), 3.75 (dd, J=20.4, 12.9 Hz, 1H), 3.60 (s, 1H), 3.52-3.37 (m, 2H), 3.17 (dt, J=112.0, 29.0 Hz, 4H), 2.80 (ddd, J=20.1, 14.4, 7.5 Hz, 2H), 2.10-1.57 (m, 3H), 0.75-0.63 (m, 4H); 13C NMR (101 MHz, DMSO-D6) δ 176.44 (s), 171.45 (s), 162.58-162.33 (m), 156.51 (s), 148.02 (s), 147.63 (s), 144.61 (s), 134.87 (s), 128.43 (s), 123.09 (s), 109.13 (s), 108.67 (s), 101.87 (d, J=4.5 Hz), 51.81 (s), 51.23 (s), 46.50 (s), 44.91 (s), 29.70-28.64 (m), 12.13 (s), 7.43 (d, J=5.9 Hz).
HRMS (ESI+) calculated for C26H27N7O4 [M+H]+: 501.5470, found 502.5154.
21e (7.1 mg, 53.6%). 1H NMR (400 MHz, DMSO-d6) δ 8.30-8.23 (m, 1H), 7.75 (d, J=22.6 Hz, 1H), 7.42 (d, J=18.5 Hz, 1H), 7.28 (s, 1H), 7.04 (d, J=8.3 Hz, 1H), 6.91 (s, 1H), 6.73 (dd, J=8.3, 2.3 Hz, 1H), 6.25 (d, J=41.6 Hz, 1H), 4.54 (dd, J=16.0, 7.7 Hz, 2H), 4.02 (dd, J=14.2, 7.1 Hz, 1H), 3.74 (dd, J=22.2, 14.9 Hz, 1H), 3.56 (d, J=30.0 Hz, 2H), 3.50-3.46 (m, 1H), 3.16 (t, J=8.4 Hz, 2H), 3.04 (dd, J=16.9, 11.2 Hz, 2H), 2.85 (dd, J=14.2, 9.4 Hz, 1H), 2.73 (dd, J=14.5, 6.0 Hz, 1H), 1.95-1.84 (m, 1H), 1.75 (dt, J=12.4, 6.2 Hz, 1H), 1.60 (s, 1H), 0.73 (dd, J=12.5, 5.9 Hz, 5H); HRMS (ESI+) calculated for C27H30N7O3 [M+H]+: 500.2405, found 500.2830.
22a (10.5 mg, 47%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.77-7.39 (m, 5H), 6.94 (s, 1H), 6.45 (d, J=46.5 Hz, 1H), 4.02 (d, J=67.0 Hz, 2H), 3.68-3.58 (m, 1H), 3.02 (s, 2H), 2.90 (dd, J=24.6, 13.9 Hz, 2H), 2.74 (t, J=25.7 Hz, 2H), 1.89 (d, J=36.8 Hz, 1H), 1.62 (s, 3H), 1.46-1.33 (m, 1H), 0.77-0.41 (m, 5H).
HRMS (ESI+) calculated for C27H28F4N7O2 [M+H]+: 558.2235, found 558.2905.
22b (20 mg, 55%)1H NMR (400 MHz, MeOD) δ 8.38-8.27 (m, 1H), 7.62 (dd, J=3.3, 2.0 Hz, 1H), 7.56-7.53 (m, 1H), 7.29 (t, J=9.1 Hz, 1H), 6.59-6.40 (m, 1H), 4.12-3.99 (m, 1H), 3.83-3.75 (m, 1H), 3.30-3.13 (m, 3H), 3.10-2.88 (m, 3H), 1.84 (s, 2H), 1.57-1.40 (m, 2H), 1.31 (d, J=7.9 Hz, 2H), 0.92 (t, J=6.9 Hz, 2H), 0.85 (s, 1H), 0.79-0.56 (m, 2H). HRMS (ESI+) calculated for C26H27Cl2N7O2 [M+H]+: 540.4490, found 540.4203.
22c (9 mg, 78%)1H NMR (400 MHz, MeOD) δ 8.25 (d, J=35.1 Hz, 1H), 7.99 (d, J=6.0 Hz, 1H), 7.93-7.82 (m, 3H), 7.59-7.51 (m, 2H), 7.42 (d, J=8.2 Hz, 1H), 6.49 (d, J=55.7 Hz, 1H), 3.79 (ddd, J=18.4, 12.7, 6.3 Hz, 2H), 3.30-3.14 (m, 2H), 3.11-2.93 (m, 3H), 2.88 (s, 1H), 1.44 (d, J=29.3 Hz, 2H), 1.33 (d, J=14.7 Hz, 4H), 0.92 (t, J=6.9 Hz, 2H), 0.83 (s, 1H), 0.66 (d, J=47.0 Hz, 2H). HRMS (ESI+) calculated for C30H31N7O2 [M+H]+: 521.6250, found 522.5953.
22d (5.6 mg, 67%). 1H NMR (400 MHz, MeOD) δ 8.24 (d, J=21.4 Hz, 1H), 6.95-6.81 (m, 3H), 6.25 (s, 1H), 6.01 (s, 2H), 3.95 (s, 1H), 3.81-3.72 (m, 1H), 3.29-2.82 (m, 6H), 1.80 (s, 2H), 1.62 (s, 2H), 1.31 (s, 2H), 0.92 (t, J=6.8 Hz, 2H), 0.87-0.81 (m, 1H), 0.67 (d, J=57.8 Hz, 2H). HRMS (ESI+) calculated for C27H29N7O4 [M+H]+: 515.5740, found 516.4862.
22e (7.3 mg, 55%)1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.43 (s, 2H), 7.25 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.73 (t, J=8.7 Hz, 1H), 6.18 (dd, J=38.9, 33.6 Hz, 1H), 4.55 (dd, J=17.2, 8.2 Hz, 2H), 4.00 (dt, J=45.6, 22.8 Hz, 2H), 3.63-3.53 (m, 1H), 3.13 (dd, J=26.9, 19.0 Hz, 5H), 2.92-2.66 (m, 3H), 1.77 (s, 3H), 1.48 (s, 2H), 0.75-0.44 (m, 5H); HRMS (ESI+) calculated for C28H32N7O3 [M+H]+: 514.2561, found 514.2889.
23a (13.8 mg, 81.9%). 1H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 7.78-7.40 (m, 5H), 6.94 (s, 1H), 6.47 (d, J=47.9 Hz, 1H), 4.16-3.89 (m, 2H), 3.61 (dd, J=15.7, 7.9 Hz, 1H), 3.12-2.66 (m, 6H), 1.99-1.88 (m, 1H), 1.62 (s, 2H), 1.47-1.33 (m, 1H), 0.79-0.33 (m, 5H); HRMS (ESI+) calculated for C27H28F4N7O2 [M+H]+: 558.2235, found 558.2545.
23b (20 mg, 74%)1H NMR (400 MHz, DMSO) δ 8.36 (s, 1H), 7.48 (dd, J=93.0, 39.3 Hz, 5H), 6.94 (s, 1H), 6.58-6.28 (m, 1H), 4.22-3.85 (m, 2H), 3.69-3.56 (m, 1H), 3.23-2.64 (m, 7H), 1.92 (d, J=12.5 Hz, 1H), 1.63 (s, 2H), 1.35 (dd, J=5.3, 3.8 Hz, 2H), 0.78-0.45 (m, 4H); MS(ESI+) calculated for C26H28Cl2N7O2 [M+H]+: 540.1676, found 540.4310.
23c (27 mg, 80%)1H NMR (400 MHz, MeOD) δ 8.25 (d, J=35.1 Hz, 1H), 7.99 (d, J=6.0 Hz, 1H), 7.93-7.82 (m, 3H), 7.59-7.51 (m, 2H), 7.42 (d, J=8.2 Hz, 1H), 6.49 (d, J=55.7 Hz, 1H), 3.79 (ddd, J=18.4, 12.7, 6.3 Hz, 2H), 3.30-3.14 (m, 2H), 3.11-2.93 (m, 3H), 2.88 (s, 1H), 1.44 (d, J=29.3 Hz, 2H), 1.33 (d, J=14.7 Hz, 4H), 0.92 (t, J=6.9 Hz, 2H), 0.83 (s, 1H), 0.66 (d, J=47.0 Hz, 2H). HRMS (ESI+) calculated for C30H31N7O2 [M+H]+: 521.6250, found 522.5953.
23d (6 mg, 74%). 1H NMR (400 MHz, DMSO-D6) δ 8.26 (s, 1H), 7.48 (d, J=39.9 Hz, 2H), 6.89 (d, J=8.2 Hz, 3H), 6.82-6.78 (m, 1H), 6.34-6.10 (m, 1H), 6.04 (s, 2H), 4.45-3.68 (m, 2H), 3.63-3.54 (m, 1H), 3.02 (s, 4H), 2.78 (d, J=45.4 Hz, 3H), 1.97 (s, 1H), 1.76 (s, 2H), 1.40 (d, J=54.2 Hz, 2H), 0.74-0.43 (m, 4H); HRMS (ESI+) calculated for C27H29N7O4 [M+H]+: 515.5740, found 516.4862.
23e (6 mg, 35.5%). 1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.43 (s, 2H), 7.26 (s, 1H), 7.02 (s, 1H), 6.90 (s, 1H), 6.73 (t, J=8.9 Hz, 1H), 6.18 (dd, J=39.9, 34.5 Hz, 1H), 4.56 (dd, J=16.9, 8.3 Hz, 2H), 4.15-3.83 (m, 2H), 3.63-3.53 (m, 1H), 3.13 (dd, J=26.9, 19.1 Hz, 5H), 2.92-2.63 (m, 3H), 1.76 (s, 3H), 1.47 (s, 2H), 0.78-0.39 (m, 5H). HRMS (ESI+) calculated for C28H32N7O3 [M+H]+: 514.2561, found 514.2899.
Synthesis was carried out using methylamine in 33% ethanol (1.75 mL) instead of an ammonia solution in the same synthesis method as above, and an organic layer was washed with water and brine. After removing water with anhydrous magnesium sulfate (MgSO4) and concentrating the solvent, the resulting solution was purified by column chromatography using MC and MeOH (MC:MeOH=20:1) as mobile phase, thereby obtaining Compound 24.
24a (14.5 mg, 83.9%). 1H NMR (400 MHz, DMSO-d6) δ 8.40-8.34 (m, 1H), 7.92 (dd, J=18.6, 6.7 Hz, 1H), 7.89-7.65 (m, 3H), 7.58-7.48 (m, 1H), 6.52 (d, J=28.1 Hz, 1H), 3.73 (d, J=6.3 Hz, 1H), 3.61 (dd, J=15.1, 7.2 Hz, 2H), 3.44-3.38 (m, 1H), 3.28-3.17 (m, 1H), 3.06 (d, J=8.1 Hz, 2H), 2.87 (dd, J=20.0, 11.0 Hz, 1H), 2.79-2.72 (m, 1H), 2.62 (d, J=4.4 Hz, 3H), 1.99-1.84 (m, 1H), 1.72 (s, 1H), 1.47 (dd, J=26.8, 19.8 Hz, 1H), 0.76-0.63 (m, 5H); HRMS (ESI+) calculated for C27H28F4N7O2 [M+H]+: 558.2235, found 558.2545.
24b (17 mg, 91%)1H NMR (400 MHz, DMSO) δ 8.37 (dd, J=13.7, 5.2 Hz, 1H), 7.94 (d, J=4.5 Hz, 1H), 7.87-7.74 (m, 1H), 7.68-7.60 (m, 2H), 7.31 (t, J=7.0 Hz, 1H), 6.60-6.36 (m, 1H), 3.74 (s, 1H), 3.67-3.40 (m, 3H), 3.27-2.69 (m, 6H), 2.62 (d, J=4.5 Hz, 3H), 2.02-1.87 (m, 1H), 1.82-1.65 (m, 2H), 0.73-0.67 (m, 4H); MS(ESI+) calculated for C26H28Cl2N7O2+[M+H]+: 540.1676, found 540.5031.
24d (16 mg, 84%). 1H NMR (400 MHz, DMSO-D6) δ 8.33-8.26 (m, 4H), 7.91 (t, J=7.6 Hz, 4H), 7.79 (d, J=22.8 Hz, 4H), 6.96-6.87 (m, 8H), 6.82 (dd, J=8.1, 1.4 Hz, 4H), 6.31 (d, J=34.3 Hz, 3H), 6.07-5.99 (m, 8H), 3.76 (dt, J=26.3, 13.2 Hz, 4H), 3.65-3.54 (m, 6H), 3.33 (s, 12H), 3.30-2.90 (m, 14H), 2.90-2.65 (m, 9H), 2.65-2.58 (m, 12H), 2.07-1.52 (m, 13H), 0.75-0.64 (m, 17H). HRMS (ESI+) calculated for C27H29N7O4 [M+H]+: 515.5740, found 516.6663.
25a (19.4 mg, 99.4%)1H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J=9.8, 5.2 Hz, 1H), 7.95 (d, J=4.5 Hz, 1H), 7.83-7.69 (m, 3H), 7.58-7.47 (m, 1H), 6.52 (d, J=28.5 Hz, 1H), 3.74 (s, 1H), 3.67-3.54 (m, 2H), 3.45-3.38 (m, 1H), 3.28-3.18 (m, 1H), 3.06 (d, J=7.7 Hz, 2H), 2.87 (dd, J=20.1, 10.9 Hz, 1H), 2.79-2.73 (m, 1H), 2.62 (d, J=4.5 Hz, 3H), 1.90 (dd, J=38.1, 15.8 Hz, 1H), 1.72 (s, 1H), 1.61-1.43 (m, 1H), 0.78-0.60 (m, 5H); HRMS (ESI+) calculated for C27H28F4N7O2 [M+H]+: 558.2235, found 558.2545.
25b (5 mg, 67%)1H NMR (400 MHz, DMSO) δ 8.39 (dd, J=13.8, 5.2 Hz, 1H), 7.97 (d, J=4.5 Hz, 1H), 7.69 (ddd, J=27.6, 18.8, 10.6 Hz, 3H), 7.33 (t, J=7.0 Hz, 1H), 6.52 (d, J=40.2 Hz, 1H), 3.77 (d, J=5.9 Hz, 1H), 3.55 (ddd, J=32.7, 17.2, 7.4 Hz, 3H), 3.28-2.72 (m, 6H), 2.64 (d, J=4.5 Hz, 3H), 1.97 (dd, J=21.1, 7.5 Hz, 1H), 1.82-1.67 (m, 2H), 0.72 (dd, J=9.8, 8.2 Hz, 4H); MS(ESI+) calculated for C26H28Cl2N7O2+ [M+H]+: 540.1676, found 540.5031.
25d (27 mg, 92%). 1H NMR (400 MHz, DMSO-D6) δ 8.33-8.26 (m, 2H), 7.78 (d, J=24.4 Hz, 2H), 7.44 (s, 2H), 6.96-6.87 (m, 6H), 6.82 (dd, J=8.1, 1.4 Hz, 2H), 6.31 (d, J=29.3 Hz, 2H), 6.08-6.00 (m, 4H), 3.74 (dd, J=20.1, 12.7 Hz, 2H), 3.60 (s, 3H), 3.45 (dd, J=32.0, 12.1 Hz, 6H), 3.28-2.99 (m, 6H), 2.79 (ddd, J=20.1, 14.3, 7.4 Hz, 4H), 2.07-1.58 (m, 6H), 0.75-0.65 (m, 8H). 13C NMR (101 MHz, DMSO-D6) δ 176.46 (s), 171.50 (s), 162.43 (d, J=2.3 Hz), 144.79-144.50 (m), 126.51 (s), 123.10 (s), 109.12 (s), 108.68 (s), 101.87 (d, J=4.7 Hz), 51.77 (s), 51.23 (s), 46.49 (s), 44.92 (s), 29.77-29.10 (m), 12.48 (s), 7.44 (d, J=5.8 Hz); HRMS (ESI+) calculated for C27H29N7O4 [M+H]+: 515.5740, found 516.5222.
26a (17.6 mg, 77%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.93 (t, J=13.5 Hz, 1H), 7.79-7.55 (m, 3H), 7.50 (s, 1H), 6.45 (d, J=48.9 Hz, 1H), 4.02 (d, J=61.1 Hz, 2H), 3.68-3.58 (m, 1H), 3.11-2.75 (m, 6H), 2.62 (t, J=4.3 Hz, 3H), 1.95 (d, J=16.3 Hz, 1H), 1.62 (d, J=5.8 Hz, 2H), 1.50-1.27 (m, 2H), 0.75-0.43 (m, 5H); HRMS (ESI+) calculated for C28H30F4N7O2 [M+H]+: 572.2392, found 572.2972.
26b (16 mg, 57%)1H NMR (400 MHz, DMSO) δ 8.35 (s, 1H), 7.94 (d, J=4.5 Hz, 1H), 7.55 (d, J=44.7 Hz, 3H), 7.27 (s, 1H), 6.41 (d, J=53.6 Hz, 1H), 3.99 (d, J=38.5 Hz, 2H), 3.66-3.55 (m, 1H), 3.25-2.66 (m, 7H), 2.62 (t, J=4.6 Hz, 3H), 1.95 (d, J=12.2 Hz, 1H), 1.63 (s, 2H), 1.37 (dd, J=23.3, 15.5 Hz, 2H), 0.76-0.47 (m, 4H); MS(ESI+) calculated for C27H30Cl2N7O2+[M+H]+: 554.1833, found 554.6538.
26c (9 mg, 87%). 1H NMR (400 MHz, DMSO-D6) δ 8.23 (s, 1H), 8.02-7.82 (m, 5H), 7.57-7.35 (m, 4H), 6.44-6.11 (m, 1H), 4.45-4.06 (m, 1H), 3.83 (s, 2H), 3.61 (dd, J=15.7, 7.8 Hz, 1H), 3.04 (d, J=45.2 Hz, 3H), 2.95-2.68 (m, 3H), 2.63 (t, J=4.0 Hz, 3H), 1.78 (dd, J=71.8, 52.5 Hz, 2H), 1.52-1.30 (m, 2H), 1.23 (s, 1H), 0.74-0.38 (m, 4H); HRMS (ESI+) calculated for C31H33N7O2 [M+H]+: 535.6520, found 536.5063.
27a (11.6 mg, 78%). 1H NMR (400 MHz, DMSO-d6) δ 8.36 (s, 1H), 7.94 (d, J=4.4 Hz, 1H), 7.65 (d, J=63.4 Hz, 3H), 7.49 (s, 1H), 6.44 (d, J=49.8 Hz, 1H), 4.02 (d, J=62.4 Hz, 2H), 3.60 (dd, J=15.4, 7.8 Hz, 1H), 2.85 (ddd, J=33.8, 32.4, 25.2 Hz, 6H), 2.62 (t, J=4.3 Hz, 3H), 1.99-1.89 (m, 1H), 1.61 (s, 2H), 1.43 (d, J=7.5 Hz, 2H), 0.77-0.41 (m, 5H); HRMS (ESI+) calculated for C28H30F4N7O2 [M+H]+: 572.2392, found 572.2972.
27b (19 mg, 86%)1H NMR (400 MHz, DMSO) δ 8.35 (s, 1H), 7.94 (d, J=4.7 Hz, 1H), 7.56 (t, J=23.0 Hz, 3H), 7.27 (s, 1H), 6.41 (d, J=54.7 Hz, 1H), 3.99 (d, J=40.1 Hz, 2H), 3.66-3.55 (m, 1H), 3.23-2.66 (m, 7H), 2.62 (t, J=4.6 Hz, 3H), 1.95 (d, J=12.6 Hz, 1H), 1.63 (s, 2H), 1.51-1.29 (m, 2H), 0.63 (dd, J=74.5, 27.0 Hz, 4H); MS(ESI+) calculated for C27H30Cl2N7O2+[M+H]+: 554.1833, found 554.4377.
27c (5.3 mg, 49%). 1H NMR (400 MHz, DMSO-D6) δ 8.23 (s, 1H), 7.95 (d, J=4.7 Hz, 1H), 7.88 (d, J=14.0 Hz, 2H), 7.73 (s, 3H), 7.57-7.50 (m, 1H), 7.40 (s, 1H), 6.26 (d, J=95.0 Hz, 1H), 3.67-3.57 (m, 1H), 3.50-3.40 (m, 3H), 2.95 (dd, J=72.1, 23.6 Hz, 3H), 2.66 (ddd, J=16.2, 12.2, 6.4 Hz, 5H), 2.00-1.84 (m, 2H), 1.31 (d, J=62.9 Hz, 3H), 0.78 (dt, J=115.4, 35.5 Hz, 4H). HRMS (ESI+) calculated for C31H33N7O2 [M+H]+: 535.6520, found 536.7582.
The synthesis methods according to Preparation Examples and Examples can be shown in Scheme 1 below.
In Scheme 1, steps are as follows: (i) OsO4, NMO, t-BuOH, THF, H2O, room temperature, 2 hrs; (ii) MsCl, TEA, MC, room temperature, 2 hrs; (iii) NaN3, DMF, 80° C., 12 hrs; (iv) Pd(OH)2, H2(g), MeOH, room temperature, 1 hr; (v) 7, aryl imidate, EtOH, 80° C., 5 hrs, (vi) oxalyl chloride, DMSO, TEA, MC, −78° C.->room temperature, 2 hrs; (vii) 10, Cs2CO3, DMF, 80° C., MW 1 hr; (viii) oxone, MeOH:H2O=1:1, room temperature, 1 hr; (ix) 12, THF, 60° C., 5 hrs; (x) 4M-HCl in dioxane, room temperature, 1 hr; (xi) cyclopropanecarbonyl chloride, THF, 60° C., 5 hrs, xii) NH3 in 7N MeOH.
Changes in JNK3 enzyme activity due to treatment with the imidazole derivatives of Tables 1 to 5 below according to the present invention were confirmed through IC50.
The IC50 values of JNK3 of the synthesized compounds were measured and summarized in Tables 1 to 5.
It should be understood by those of ordinary skill in the art that the above description of the present invention is exemplary, and the exemplary embodiments disclosed herein can be easily modified into other specific forms without departing from the technical spirit or essential features of the present invention. Therefore, the exemplary embodiments described above should be interpreted as being illustrative and not restrictive in any aspect.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2021-0108424 | Aug 2021 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2022/012173 | 8/16/2022 | WO |
