Aromatic compounds, their preparation methods and applications

Abstract

The present invention discloses an aromatic compound, its preparation method and its use.The present invention provides a compound represented by formula I or a pharma- ceutically acceptable salt thereof.The compound of the present invention shows good safety, tolerability and reduction of liver fat in mice, and has shown potential efficacy in treating NASH in animal experiments of NASH disease model. [Formula 1] JPEG2024510431000231.jpg6266

Description

[Technical Field]

[0001] This application claims priority to Chinese Patent Application No. 2021102648447, filed on 4 March 2021. This application incorporates the full text of said Chinese Patent Application. This invention belongs to the field of chemical pharmaceuticals, and more specifically relates to aromatic compounds, methods for preparing them, and their applications. [Background technology]

[0002] The thyroid gland is a butterfly-shaped organ located at the base of the neck. It secretes hormones known as thyroid hormones, which control the body's basal metabolism and how our bodies use energy. Thyroid hormones regulate important bodily functions such as respiration, heart rate, the central and peripheral nervous systems, body weight, muscle strength, menstrual cycle, body temperature, and cholesterol levels.

[0003] Thyroid hormone receptors (THR) belong to a superfamily of nuclear receptors whose expression is induced by the thyroid hormone T3. The major subtypes of thyroid hormone receptors, THRα-1, THRβ-1, and THRβ-2, primarily mediate the action of thyroid hormones essential for human growth and metabolism. THRβ-1 is widely expressed in all tissues, but more prominently in the brain, thyroid gland, liver, and kidneys, while THRβ-2 is tissue-specific, primarily expressed in the anterior pituitary gland, pituitary gland, retina, developing brain, and inner ear.

[0004] The physiological effects of thyroid hormones affect almost all organ systems. Clinically, these effects manifest as changes in lipid metabolism and impacts on cardiovascular development. Thyroid hormones have beneficial effects such as lowering cholesterol, improving lipid profiles, and treating obesity. Thyroid hormone analogs can improve lipid profiles by lowering low-density lipoprotein (LDL) cholesterol, increasing the reuptake of high-density lipoprotein (HDL) cholesterol, stimulating reverse cholesterol transport, and lowering plasma triglycerides.

[0005] Since THRβ-1 is the major thyroid hormone receptor isoform in the liver, it is hypothesized that inhibition of normal THRβ-1 activity by its mutants leads to metabolic abnormalities. Furthermore, thyroid hormones also regulate apolipoprotein B, the major protein component of very low-density lipoprotein (VLDL). Several findings suggest that thyroid hormones not only stimulate the lipid oxidation pathway but also inhibit the pathway by which lipid droplets act as lipid storage organelles, thereby promoting the secretion of lipid droplets as part of VLDL liposomes.

[0006] This study showed that patients with low thyroid function had a significantly higher rate of non-alcoholic steatohepatitis (NASH) and advanced fibrosis compared to patients with completely normal thyroid function (NASH: 52.4% vs. 37.2%, advanced fibrosis: 21.0% vs. 10.6%, P<0.01). Furthermore, patients with subclinical hypothyroidism had a significantly higher incidence of NASH and advanced fibrosis compared to patients with low thyroid function (NASH: 57.6% vs. 48.8%; advanced fibrosis: 25.4% vs. 17.9%; P < 0.01). Serum thyroid-stimulating hormone levels were significantly higher in NASH patients compared to the normal group. In addition, thyroid function tests confirmed the presence of several thyroid abnormalities in patients with chronic liver disease. Furthermore, hypothyroidism was not associated with NASH but was associated with other known metabolic risk factors; i.e., hypothyroidism is an independent risk factor for NASH.

[0007] If the harmful effects of thyroid hormone excess can be separated from its potential beneficial effects on cholesterol and lipid reduction, there is hope for the development of new drugs with potent effects. Considering the roughly three stages of progression in non-alcoholic fatty liver disease (NAFLD) and NASH: namely, fat deposition, hepatitis-hepatocyte death / apoptosis, and fibrosis / cirrhosis, treatment strategies should include at least three aspects: namely, reduction or removal of hepatic fat deposition, control and suppression of persistent hepatitis in the hepatic region / reduction of hepatocyte death, and degradation of fibrous / extracellular matrix formed to halt the progression of fibrosis or reverse the fibrotic process. Degradation of the extracellular matrix to reverse the fibrotic process. Of these three aspects, the reduction or removal of fat deposition and control and suppression of ongoing inflammation / reduction of hepatocyte death are the most important. NAFLD / NASH is essentially metabolic syndrome and is closely related to lipid metabolism disorders, insulin resistance / type 2 diabetes, etc., and at the same time, these metabolic disorders are also closely related to fat deposition and type 2 diabetes. Simultaneously, these metabolic disorders and fat deposition trigger inflammation, which in turn causes hepatocyte death / apoptosis, and hepatocyte death / apoptosis naturally progresses to fibrosis / cirrhosis. Most drugs currently in Phase III clinical trials have a pharmacological mechanism of action that reduces or eliminates fat deposition / degeneration, controls and suppresses persistent inflammation, and reduces hepatocyte death.

[0008] In summary, the development of selective oral small molecule agonists targeting THR-β is a very promising strategy, and there are already two international pioneers in this field. One of them, MGL-3196, a drug candidate from Madrigal Pharmaceuticals (USA), showed promising results in a Phase II clinical trial in patients with biopsy-verified non-alcoholic steatohepatitis (NASH). MGL-3196 is a once-daily oral selective agonist targeting the liver-specific thyroid hormone receptor subtype (THR-β). In the clinical trial, the primary clinical endpoint was a statistically significant reduction of ≥30% in liver lipids, measured using MRI-PDFF (non-invasive imaging), which correlated highly with improvement in NASH on liver biopsy. In the MGL-3196 treatment group, statistically significant decreases in ALT and AST were observed, and several other secondary endpoints, including LDL-C, triglycerides, apolipoprotein B, and lipoprotein a, also showed statistically significant improvements. All of these clinical indicators correlated with the clinical status of NASH patients.

[0009] Furthermore, Viking Therapeutics has developed VK2809 (or MB07811), an oral small molecule agonist with selectivity for the thyroid hormone receptor β subtype (THR-β). This drug is currently in Phase II clinical trials in patients with primary hypercholesterolemia and non-alcoholic fatty liver disease. As a result, patients treated with VK2809 showed a significant reduction of 20% or more in LDL-C levels. In NAFLD patients, LDL-C levels were significantly reduced and liver fat volume improved after 12 weeks of treatment.

[0010] Due to the remarkable inhibitory effects of MGL-3196 and VK2809 on fat deposition / shedding, the industry predicts that thyroid hormone receptor β subtype (THR-β) agonists will be very promising in the treatment of NAFLD / NASH and metabolic syndrome.

[0011] [ka] However, currently reported THR-β agonists still have limitations, and it is crucial to improve the agonist activity and subtype selectivity of compounds, especially against the THR-α subtype. Compounds currently under investigation still have deficiencies in agonist activity and selectivity. Only when these bottlenecks are overcome can these compounds be expected to become groundbreaking new therapeutic agents for NASH and related liver diseases. [Overview of the project]

[0012] The technical problem that this invention aims to solve is the shortcomings of existing single-structure thyroid hormone receptor THR-β selective agonists. Therefore, this invention provides a type of aromatic compound, a method for preparing the same, and its applications. These agonists exhibit significantly stronger agonist activity against THR-β than the currently studied drug MGL-3196, and are also significantly more selective against the THR-α subtype than MGL-3196. In animal studies using a NASH disease model, certain compounds showed good safety, tolerability, hepatic steatosis-reducing effects, and hepatoprotective effects in mice.

[0013] The present invention provides a compound represented by formula I having the structure shown below, or a pharmaceutically acceptable salt thereof:

[0014] [ka]

[0015] Here, A is either O or CH2; M is [ka] or [ka] That is the case.

[0016] X and Y are, independently, chlorine, bromine, and iodine, "I-1 isotope". 124I or 131 It is either "I" or a C1-C6 alkyl group; R1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, "one or more fluorine-substituted C1-C6 alkyl", "one or more fluorine-substituted C2-C6 alkenyl", "one or more deuterium-substituted C1-C6 alkyl", or "one or more deuterium-substituted C2-C6 alkenyl"; R 2 This refers to C2-C6 alkenyls, "one or more fluorosubstituted C1-C6 alkyls", "one or more fluorosubstituted C2-C6 alkenyls", "one or more deuterium substituted C1-C6 alkyls", or "one or more deuterium substituted C2-C6 alkenyls".

[0017] In some embodiments, certain substituents in the compound of formula I or its pharmaceutically acceptable salt may have further definitions, and substituents not covered below are defined as described in any embodiment of the present invention (hereinafter referred to as "in some embodiments").

[0018] Here, A is either O or CH2; X and Y are independently chlorine, bromine, iodine, or C1-C6 alkyl; M is [ka] That is the case. R1 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, one or more fluorine-substituted C1-C6 alkyl, one or more fluorine-substituted C2-C6 alkenyl, one or more deuterium-substituted C1-C6 alkyl, or one or more deuterium-substituted C2-C6 alkenyl; Alternatively, M is [ka] And; R 2 This is "one or more fluorosubstituted C1-C6 alkyl groups". In one embodiment, A is O; X and Y are independently chlorine, bromine, or iodine; and M is [Chemical formula] and R 1 is C2 - C6 alkenyl or "C1 - C6 alkyl substituted with one or more fluorines".

[0019] In one embodiment, A is O, X and Y are independently chlorine or bromine; M is [Chemical formula] and R 1 is C2 - C6 alkenyl or "fluoro - substituted C1 - C6 alkyl".

[0020] In one embodiment, X and Y are independently chlorine, bromine, iodine or CH3.

[0021] In one embodiment, R 1 is C2 - C6 alkenyl, or "C1 - C6 alkyl substituted with one or more fluorines".

[0022] In one embodiment, R 2 is "C1 - C6 alkyl substituted with one or more fluorines".

[0023] In one embodiment, when R 1 is a C2 - C6 alkenyl group, the C2 - C6 alkenyl group is a C2 - C4 alkenyl group; For example, [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [ka] [ka] or [ka] And; also, for example, [ka] That is the case.

[0024] In one embodiment, R 1 If the C1-C6 alkyl group is C1-C6 alkyl; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; preferably isopropyl.

[0025] In one embodiment, R 1 If is "one or more fluorine-substituted C1-C6 alkyls", then the C1-C6 alkyls are C1-C4 alkyls; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and "one or more" is one or three. [ka] or [ka] That is the case.

[0026] In one embodiment, R 1If is "one or more deuterium-substituted C1-C6 alkyls", then the C1-C6 alkyls are C1-C4 alkyls; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and "one or more" is one or three. [ka] That is the case.

[0027] In one embodiment, R 1 If is "one or more fluorosubstituted C2-C6 alkenyls", then the C2-C6 alkenyls are C2-C4 alkenyls; for example, [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] And; the aforementioned “one or more” is one or three.

[0028] In one embodiment, R 1 If is "one or more deuterium-substituted C2-C6 alkenyls", then the C2-C6 alkenyls are C2-C4 alkenyls; for example, [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] And; the aforementioned “one or more” is one or three.

[0029] In one embodiment, R 2 If is a C2-C6 alkenyl group, then the C2-C6 alkenyl group is a C2-C4 alkenyl group; for example, [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] And; also, for example, [ka] That is the case.

[0030] In one embodiment, R 2 If is "one or more fluorine-substituted C1-C6 alkyls", then the C1-C6 alkyls are C1-C4 alkyls; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and "one or more" is one or three. [ka] or [ka] That is the case.

[0031] In one embodiment, R 2 If is "one or more deuterium-substituted C1-C6 alkyls", then the C1-C6 alkyls are C1-C4 alkyls (for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl); and "one or more" is one or three. [ka] That is the case.

[0032] In one embodiment, R 2 If is "one or more fluorosubstituted C2-C6 alkenyls", then the C2-C6 alkenyls are C2-C4 alkenyls; for example, [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] And; the aforementioned “one or more” is one or three.

[0033] In one embodiment, R 2 If is "one or more deuterium-substituted C2-C6 alkenyls", then the C2-C6 alkenyls are C2-C4 alkenyls; for example, [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] And; the aforementioned “one or more” is one or three.

[0034] In one embodiment, when X is a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C4 alkyl group; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl; preferably methyl.

[0035] In one embodiment, when Y is a C1-C6 alkyl group, the C1-C6 alkyl group is a C1-C4 alkyl group; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl; preferably methyl.

[0036] In a certain program, R 1 teeth [ka] [ka] [ka] [ka] or [ka] That is the case.

[0037] In one program, the following basis [ka] teeth, [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] That is the case.

[0038] In one embodiment, the compound represented by formula I is one of the following: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] .

[0039] In the present invention, the compounds represented by formula I, or pharmaceutically acceptable salts thereof, can be synthesized by methods similar to those known in the chemical field, the steps and conditions of which can refer to those of similar reactions in the art, and in particular, as described herein. Starting materials can typically be derived from commercially available sources or readily prepared using methods well known to those skilled in the art (available via the online databases SciFinder and Reaxys).

[0040] The present invention also provides a method for preparing a compound represented by formula I, the method comprising the following steps: in a solvent, in the presence of a base, the compound represented by formula II-a is subjected to the following ring-closing reaction to obtain the compound represented by formula I, namely; wherein the formula, M, X, Y and A are defined as described above, and R 6 It is a C1-C6 alkyl group; [ka]

[0041] In one embodiment, R 6 If the C1-C6 alkyl group is C1-C4 alkyl; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, butyl, tert-butyl, isobutyl, sec-butyl, or tert-butyl; for example, ethyl.

[0042] The present invention also provides a compound represented by formula II-a. [ka]

[0043] Here, M, A, X, Y, and R 6 It is defined as described above.

[0044] In one embodiment, the compound represented by formula II-a is one of the following compounds: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or [ka] .

[0045] The present invention also provides a method for preparing the compound represented by formula II-a. This method comprises the steps of reacting the compound represented by formula II-b with sodium nitrite in a solvent in the presence of an acid, and then reacting it with cyanoacetylcarbamic acid in the reaction shown in the following formula to obtain the compound represented by formula II-a; where M, A, X, Y and R 6 It is defined as described above; [ka] .

[0046] The present invention provides a pharmaceutical composition comprising substance A and one or more pharmaceutically acceptable carriers; substance A is, as previously described, a compound represented by formula I or a pharmaceutically acceptable salt thereof. In the pharmaceutical composition, the amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof may be a therapeutically effective amount.

[0047] The pharmaceutically acceptable carriers (pharmaceutical excipients) described may be those widely used in the field of pharmaceutical manufacturing. Excipients are primarily used to provide safe, stable, and functional pharmaceutical compositions and may also provide means for dissolving the active ingredient at a desired rate after administration to a subject, or means for promoting the efficient absorption of the active ingredient after administration to a subject. The pharmaceutical excipient may be an inert filler, or it may provide some function such as stabilizing the pH of the entire composition or preventing the decomposition of the active ingredient of the composition. The pharmaceutical excipient may include one or more of the following: binders, suspension aids, emulsifiers, diluents, fillers, granulators, adhesives, disintegrants, lubricants, anti-tacks, flow aids, wetting agents, gelling agents, absorption retarders, dissolution inhibitors, reinforcing agents, adsorbents, buffers, chelating agents, preservatives, colorants, flavor modifiers, and sweeteners.

[0048] The pharmaceutical compositions of the present invention can be prepared in accordance with this disclosure using any method known to those skilled in the art. Examples include conventional mixing, dissolution, granulation, emulsification, grinding, encapsulation, embedding, or freeze-drying processes.

[0049] The present invention also provides the application of substance B in the preparation of THR-β agonists, wherein substance B is a compound as represented by formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above.

[0050] In the applications described, the THR-β agonist may be used in mammalian organisms. It may also be used in non-biological organisms, primarily for experimental purposes, for example, as a standard or control sample to provide comparison, or as a kit by conventional methods of the art to provide a rapid assay for the agonist activity of THR-β.

[0051] The present invention also provides the application of substance B in the preparation of a pharmacopoeia for the treatment and / or prevention of THR-β-related diseases, wherein substance B is a compound represented by formula I above or a pharmaceutically acceptable salt thereof, or the pharmacopoeia described above.

[0052] The listed conditions are one or more selected from non-alcoholic fatty liver disease, obesity, hepatic fibrosis, type 2 diabetes, and primary hypercholesterolemia.

[0053] Unless otherwise specified, the following terms appearing in the specification and claims of this invention have the meanings set forth below: The term "pharmaceutically acceptable" means that the salts, solvents, excipients, etc., are generally non-toxic, safe, and suitable for use in patients. The "patient" is preferably a mammal, and more preferably a human.

[0054] The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention prepared using a relatively non-toxic, pharmaceutically acceptable acid or base. If the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting a prototype of such a compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include, but are not limited to, lithium salts, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, zinc salts, bismuth salts, ammonium salts, and diethanolamine salts. If the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting a prototype of such a compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Examples of pharmaceutically acceptable acids include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, and sulfuric acid. Examples of pharmaceutically acceptable acids include organic acids, such as acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, octanedioic acid, transbutylenedioic acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, saccharic acid, formic acid, ethanesulfonic acid, bis-hydroxynaphthoic acid (i.e., 4,4'-methylene-bis(3-hydroxy-2-naphthalenecarboxylic acid)), and amino acids (e.g., glutamic acid, arginine). If the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts.See, in particular, Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science 66:1-19 (1977), or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0055] In the definition of a compound, any variable (e.g., R) 1 When a variable appears multiple times, the definition of each position in which that variable appears is independent of the definitions of the remaining positions in which the variable appears, and their meanings are independent of each other and do not influence each other. Therefore, if a given R has one, two, or three elements... 1 If substituted with a group, i.e., the group has up to 3 R 1 If substitution is acceptable, then R at that position 1 The definition is R at the remaining position 1 It is independent of the definition. Furthermore, combinations of substituents and / or variables are permitted only if the combination results in a stable compound.

[0056] In this specification, certain chemical groups are preceded by a simplified symbol indicating the total number of carbon atoms present in that group. For example, C1-C6 alkyl refers to the alkyl groups defined below, having a total of 1, 2, 3, 4, 5, or 6 carbon atoms. The total number of carbon atoms in the simplified notation does not include any carbon atoms that may be present in substituents of the group.

[0057] "Treatment" means a therapeutic procedure. In relation to a specific pathological condition, treatment means (1) alleviating one or more biological manifestations of a disease or pathological condition; (2) (a) inhibiting one or more points in a biological cascade that causes or contributes to a pathological condition, or (b) inhibiting one or more biological manifestations of a pathological condition; (3) improving one or more symptoms, effects, or side effects associated with a pathological condition, or one or more symptoms, effects, or side effects associated with the pathological condition or its treatment; or (4) slowing the progression of a pathological condition or one or more biological manifestations of a pathological condition. (4) Delaying the progression of a disease or the biological manifestation of one or more disease conditions.

[0058] "Therapeutic dose" means an amount of the compound sufficient to effectively treat the disease or condition described herein when administered to a patient. The therapeutic dose varies depending on the compound, the condition and its severity, and the age of the patient being treated, and may be adjusted as needed by those skilled in the art.

[0059] As will be understood by those skilled in the art, in accordance with the conventions used in the art, in the structural formulas describing parts in this application, [ka] Using this symbol means that the corresponding site is linked to other fragments or sites in the compound through this site.

[0060] Provided that it does not contradict common sense in the field, the above preferred conditions can be arbitrarily combined to obtain each preferred embodiment of the present invention.

[0061] The reagents and raw materials used in this invention are commercially available.

[0062] The positive progressive effects of this invention are as follows: This invention provides a thyroid hormone receptor agonist, a method for preparing the same, and its applications. This agonist has significantly stronger agonist activity against THR-β than the currently investigated drug MGL-3196, and exhibits significantly higher selectivity for the THR-α subtype than MGL-3196. In animal experiments using a NASH disease model mouse, this agonist showed good safety, tolerability, and hepatic steatosis-reducing effects, and is expected to have a therapeutic effect on NASH. [Brief explanation of the drawing]

[0063] [Figure 1] Figure 1 shows the body weight change curve of animals that were administered effective sample 3 for 42 days; [Figure 2] Figure 2 shows the total cholesterol (TCHO) value in the third efficacy embodiment; [Figure 3] Figure 3 shows the low-density lipoprotein (LDL) values ​​in Effect Embodiment 3; [Figure 4] Figure 4 shows the hepatocyte balloon-like change score for effector embodiment 3; [Figure 5] Figure 5 shows the liver inflammation score for case 3, which showed positive results; [Figure 6] Figure 6 shows example 3 of the effect of the liver fibrosis (Ishak) score. [Figure 7] Figure 7 shows the liver NAS score of the affected Example 3. [Figure 8] Figure 8 shows the body weight change curve of animals administered with Efficacy Embodiment 4 over a 3-day period. [Modes for carrying out the invention]

[0064] The present invention will be further described below with reference to embodiments, but this will not limit the present invention to the scope of the embodiments described. Experimental methods for which specific conditions are not shown in the following embodiments will be selected according to conventional methods and conditions or the descriptions of those skilled in the art. [Examples]

[0065] Example 1: 2-(3,5-dichloro-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 1) [ka]

[0066] Step 1: 2-Isopropenylbenzene-1,4-diol

[0067] 5.0 g, 26.5 mmol of 2-bromobenzene-1,4-diol, 11.0 g, 79.4 mol (20 mL) of aqueous potassium carbonate solution, 6.67 g, 39.7 mmol of isopropenylboronic acid pinacol ester, and 0.97 g, 1.32 mmol of Pd(dppf)Cl2 were added. The mixture was heated to 105°C and stirred at 105°C for 5 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove 1,4-dioxane. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% aqueous hydrochloric acid solution. The mixture was extracted with ethyl acetate (30 mL x 3) and the organic phases were combined. The organic phases were washed with saturated aqueous sodium chloride solution (10 mL). The organic phases were dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain the crude product of 2-isopropenylbenzene-1,4-diol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10:1 to 5:1) to obtain 2.50 g of the target product in 63% yield. 1 H NMR (400 MHz, CDCl3): δ 6.80 (d, J = 9.2 Hz, 1H), 6.66-6.64 (m, 2H), 5.39 (t, J = 1.6 Hz, 1H), 5.30 (s, 1H), 5.14 (s, 1H).1H), 5.14 (s, 1H), 4.50 (s, 1H), 2.09 (s, 3H).

[0068] Step 2: 4-(2,6-dichloro-4-nitrophenoxy)-2-(isopropenyl)phenol

[0069] 50 mL of acetonitrile was mixed with 2-isopropenylbenzene-1,4-diol (3.00 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-dichloro-2-fluoro-5-nitrobenzene (3.00 g, 14.3 mmol). The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid aqueous solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride aqueous solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to dryness to obtain the crude product of 4-(2,6-dichloro-4-nitrophenoxy)-2-(isopropenyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 to 15:1) to obtain 2.85 g of the product in a yield of 59%. 1 H NMR (400 MHz, CDCl3): δ 8.29 (s, 2H), 6.85 (d, J = 8.8 Hz, 1H), 6.67 (d, J = 3.2 Hz, 1H), 6.60 (dd, J = 8.8 , 3.2 Hz, 1H), 6.60 (dd, J = 8.8 , 3.2 Hz, 1H).3.2 Hz, 1H), 5.46 (s, 1H), 5.41 (t, J = 1.2 Hz, 1H), 5.17 (s, 1H), 2.08 (s, 3H).

[0070] Step 3: 4-(4-amino-2,6-dichlorophenoxy)-2-(isopropenyl)phenol

[0071] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dichloro-4-nitrophenoxy)-2-(isopropenyl)phenol (2.85 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.42 g of product in 93% yield. [M+H] + : 310.1 . 1 H NMR (400 MHz, CDCl3): δ6.81 (d, J = 8.8 Hz, 1H), 6.68-6.67 (m, 3H), 6.59 (dd, J = 8.8, 3.2 Hz, 1H), 5.38 (t, J = 1.2 Hz, 1H), 5.36 (s, 1H), 5.15 (s, 1H), 3.72 (br s, 2H)), 2.08 (s, 3H).

[0072] Step 4: Ethyl (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate

[0073] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dichlorophenoxy)-2-(isopropenyl)phenol (2.40 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.7 g of the product in 73% yield. [M+H] + : 477.2.

[0074] Step 5: 2-(3,5-dichloro-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0075] Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazone)acetyl)carbamate ethyl ester (2.70 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% aqueous ammonium bicarbonate = 20%~80%) to obtain 1.05 g of product in 43% yield. [MH] -: 428.8 .1H NMR (400 MHz, DMSO-d6): δ 13.29 (br s, 1H), 9.34 (s, 1H), 7.78 (s, 2H), 6.77 (d, J = 8.8 Hz, 1H), 6.66 (d, J = 3.2 Hz, 1H), 6.54 (dd, J = 8.8, 3.2 Hz, 1H), 5.10 (dd, J = 8.8, 3.2 Hz, 1H).1H), 5.10 (s, 2H), 2.05 (s, 3H). [Examples]

[0076] Example 2: 2-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxo-1,6-dihydropyridazine-3-yl)oxy)phenyl-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 2)

[0077] [ka]

[0078] Step 1: 3,6-Dichloro-4-(2-fluoropropan-2-yl)pyridazine

[0079] To a mixture of concentrated sulfuric acid (17.4 g, 177 mmol) and water (110 mL), 3,6-dichloropyridazine (17.6 g, 118 mmol) and 2-fluoro-2-methylpropanoic acid (25 g, 236 mmol) were added and heated to 40°C with stirring for 5 minutes. Silver nitrate (2 g, 11.8 mmol) was added and heated to 62°C, and a mixture of ammonium persulfate (45.8 g, 210 mmol) and water (220 mL) was added dropwise. A mixture of ammonium disulfate (45.8 g, 210 mmol) and water (220 mL) was added and heated to 80°C, where the mixture was allowed to react for 1 hour. After the reaction, the reaction mixture was cooled to 0-15°C, ammonia (approx. 80 mL) was added dropwise to adjust the pH of the system to 9, isopropyl ether (200 mL) extract was added, the extract was washed with sodium bisulfite (100 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The filtrate was concentrated to dryness under reduced pressure, and the concentrate was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 10:1) to obtain 3,6-dichloro-4-(2-fluoropropan-2-yl)pyridazine (15.9 g, yield 64.9%). MS (ESI) m / z: 209.0 [M + H] + .

[0080] Step 2: 3,5-Dichloro-4-((6-chloro-5-(2-fluoropropan-2-yl)pyridazine-3-yl)oxy)aniline

[0081] 3,6-Dichloro-4-(2-fluoropropan-2-yl)pyridazine (7.8 g, 37.5 mmol), 4-amino-2,6-dichlorophenol (6.67 g, 27.5 mmol), potassium carbonate (20.7 g, 150 mmol), and CuI (4.27 g, 22.5 mmol) were added to DMSO (78 mL), and the reaction was heated to 90°C; the reaction was stirred at 90°C for 17 hours. After the reaction was complete, it was cooled to room temperature. Water (500 mL) and ethyl acetate (500 mL) were added to the reaction system and separated; the organic phase was washed with saturated sodium chloride and dried over anhydrous magnesium sulfate; filtered, and the filtrate was concentrated under reduced pressure until dry; the concentrate was purified by column chromatography on silica gel to obtain crude 3,5-dichloro-4-((6-chloro-5-(2-fluoropropan-2-yl)pyridazin-3-yl)oxyaniline (12.5 g). MS (ESI) m / z: 350.0 [M + H] + .

[0082] Step 3: N-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxo-1,6-dihydropyridazine-3-yl)oxy)phenyl)acetamide

[0083] 3,5-Dichloro-4-((6-chloro-5-(2-fluoropropan-2-yl)pyridazin-3-yl)oxy)aniline (12.4 g, 35.8 mL) and sodium acetate (10.3 g, 125 mmol) were added to glacial acetic acid (170 mL), heated to 100°C, and stirred at 100°C for 16 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, and the pH of the system was adjusted to 8-9 with 1M sodium hydroxide aqueous solution; ethyl acetate (100 mL) extract was added, the organic phase was dried over anhydrous magnesium sulfate; filtered, and the filtrate was concentrated under reduced pressure until dry; the concentrate was subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 5:1-2:1) to obtain N-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxo-1,6-dihydropyridazine-3-yl)oxy)phenyl)acetamide (2.28 g, 16.3% yield in two steps). MS (ESI) m / z: 373.1 [M + H] + .

[0084] Step 4: 6-(4-amino-2,6-dichlorophenoxy)-4-(2-fluoropropan-2-yl)pyridazine-3(2H)-one

[0085] N-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxo-1,6-dihydropyridazin-3-yl)oxy)phenyl)acetamide (1.0 g, 2.7 mmol) and 6N hydrochloric acid (24 mL) were added to ethanol (20 mL) and heated to 70 °C. After the reaction was complete, the reaction mixture was cooled to room temperature. After filtration, the filtration cake was washed with water, and the solid was dried under reduced pressure to obtain 6-(4-amino-2,6-dichlorophenoxy)-4-(2-fluoropropan-2-yl)pyridazin-3(2H)-one (749 mg, yield: 84%). MS (ESI) m / z: 331.1 [M + H] + .

[0086] Step 5 Ethyl (2-cyano-2-(2-(3,5-dichloro-4-((5-(2-fluoropropyl-2-yl)-6-oxo-1,6-dihydropyridazine-3-yl)oxy)phenyl)hydrazino)acetyl)carbamate

[0087] 6-(4-amino-2,6-dichlorophenoxy)-4-(2-fluoropropan-2-yl)pyridazin-3(2H)-one (500 mg, 1.51 mmol) was added to 22.5 mL of 4N hydrochloric acid aqueous solution, cooled to 0°C, and 7.5 mL of 0.26N sodium nitrite aqueous solution was added dropwise. Stirring was continued for 2 hours after addition. 5-dichloro-4-((5-(2-fluoropropyl-2-yl)-6-oxo-1,6-dihydropyridazin-3-yl)oxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl (218 mg, yield 29%) was obtained. MS (ESI) m / z: 498.1 [M + H] + .

[0088] "Step 6" "2-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxa-1,6-dihydro-1,2,4-triazine-3-yl)oxy)phenyl-3,5-dioxa-2,3,4,5-tetrahydro-1,2,4-triazine-6-nitrile"

[0089] In a reaction flask, glacial acetic acid (4.8 mL), ethyl (2-cyano-2-(2-(3,5-dichloro-4-((5-(2-fluoropropan-2-yl)-6-oxo-1,6-dihydropyridazine-3-yl)oxy)phenyl)hydrazinyl)acetyl)carbamate (254 mg, 0.5 mmol), and sodium acetate (208 mg, 2.5 mmol) were added. The mixture was heated to 120°C and stirred at 120°C for 2 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: aqueous solution = 20%~95%) to obtain 55 mg of the product in yield 24.2%. MS (ESI) m / z: 452.0 [M + H] +. 1H NMR (400 MHz, DMSO-d6): δ 7.79 (s, 2H), 7.50 (s, 1H), 1.74 (s, 3H), 1.68 (s, 3H). 1.68 (s, 3H).

Example

[0090] Example 3: 2-(3,5-Dichloro-4-(3-(2-fluoropropan-2-yl)-4-hydroxyphenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 3)

Chem.

[0091] Step 1 4-(2,6-Dichloro-4-nitrophenoxy)-2-(2-fluoropropan-2-yl)phenol

[0092] To acetonitrile (50 mL), 2-(2-fluoropropan-2-yl)benzene-1,4-diol (3.00 g, 17.6 mmol), sodium carbonate (6.66 g, 63 mmol), and 1,3-dichloro-2-fluoro-5-nitrobenzene (2.65 g, 12.6 mmol) were added. The mixture was heated to 50 °C and stirred at 50 °C for 8 hours. After completion of the reaction, the reaction solution was cooled to room temperature and concentrated under reduced pressure to remove acetonitrile. Water (50 mL) and ethyl acetate (50 mL) were added to the concentrate, and the pH was adjusted to 2 - 3 with 10% aqueous hydrochloric acid. The mixture was extracted with ethyl acetate (30 mL×3), and the organic phases were combined. The organic phase was washed with saturated aqueous sodium chloride (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain a crude product of 4-(2,6-dichloro-4-nitrophenoxy)-2-(2-fluoropropan-2-yl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 - 15:1) to give 3.2 g, yield 70%. MS(ESI) m / z: 361.0 [M+H] + .

[0093] Step 2 4-(4-Amino-2,6-dichlorophenoxy)-2-(2-fluoropropan-2-yl)phenol

[0094] Add tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dichloro-4-nitrophenoxy)-2-(2-fluoropropan-2-yl)phenol (3.0 g, 8.38 mmol), an aqueous solution of ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) to a reaction flask. Heat the mixture to 70 °C and stir at 70 °C for 3 hours. After completion of the reaction, cool the reaction solution to room temperature. Add water (50 mL), a saturated aqueous sodium hydrogen carbonate solution (30 mL), and ethyl acetate (50 mL) to the reaction solution, stir for 10 minutes, and then filter. Extract the filtrate with ethyl acetate (20 mL × 3), and combine the organic phases. Wash the organic phase with saturated sodium chloride (50 mL), dry over anhydrous magnesium sulfate, and filter. Concentrate and dry the filtrate under reduced pressure to obtain 2.35 g of the product in a yield of 85%. MS (ESI) m / z: 331.1 [M + H] + .

[0095] Step 3 Ethyl (2-cyano-2-(2-(3,5-dichloro-4-(3-(2-fluoropropan-2-yl)-4-hydroxyphenoxy)phenyl)hydrazino)acetyl)carbamate Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dichlorophenoxy)-2-(2-fluoropropan-2-yl)phenol (2.30 g, 6.96 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.55 g of product in 72% yield. MS (ESI) m / z: 497.4 [M + H] + .

[0096] Step 4: 2-(3,5-dichloro-4-(3-(2-fluoropropan-2-yl)-4-hydroxyphenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0097] Glacial acetic acid (30 mL), ethyl (2-cyano-2-(2-(3,5-dichloro-4-(3-(2-fluoropropan-2-yl)-4-hydroxyphenoxy)phenyl)hydrazinyl)acetyl)carbamate (2.50 g, 5.03 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to a reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% aqueous ammonium bicarbonate = 20%~80%) to obtain 1.02 g of product in 45% yield. MS (ESI) m / z: 452.3 [M + H] + . 1HNMR (400 MHz, DMSO-d6): δ 13.24 (br s, 1H), 7.79 (s, 2H), 7.08 (d, 1H), 7.08 (d, 1H).7.08 (d, 1H), 7.05 (d, 1H), 6.83 (dd, 1H), 1.70 (d, 3H), 1.68 (d, 3H). [Examples]

[0098] Example 4: 2-(3,5-diiodo-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 4) [ka]

[0099] Step 1: 4-(2,6-diiodo-4-nitrophenoxy)-2-(isopropenyl)phenol

[0100] 50 mL of acetonitrile was mixed with 2-isopropenylbenzene-1,4-diol (3.00 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-diiodo-2-fluoro-5-nitrobenzene (5.62 g, 14.3 mmol). The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain the crude product of 4-(2,6-diiodo-4-nitrophenoxy)-2-(isopropenyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1~15:1) to obtain 4.86 g in yield of 65%. MS (ESI) m / z: 524.1 [M + H] + .

[0101] Step 2: 4-(4-amino-2,6-diiodophenoxy)-2-(isopropenyl)phenol

[0102] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-diiodo-4-nitrophenoxy)-2-(isopropenyl)phenol (4.38 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 3.72 g of product in 90% yield. MS (ESI) m / z: 494.1 [M + H] + .

[0103] Step 3 (2-Cyano-2-(2-(3,5-Diiodo-4-(4-Hydroxy-3-Isopropenylphenoxy)phenyl)Hydrazinylidene)Acetyl)Carbamate Ethyl

[0104] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-diiodophenoxy)-2-(isopropenyl)phenol (3.82 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 3.47 g of product in 68% yield. MS (ESI) m / z: 661.2 [M + H] + .

[0105] Step 4: 2-(3,5-diiodo-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0106] Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-diiodo-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl ester (3.74 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.91 g of product in 55% yield. MS (ESI) m / z: 615.1 [M + H] + . 1 H NMR (400 MHz, DMSO-d6): δ 13.10 (br s, 1H), 9.50 (s, 1H), 7.93 (s, 2H).7.93 (s, 2H), 7.15 (m, 2H), 6.90 (s, 1H), 5.10 (d, 2H), 2.43 (s, 1H). [Examples]

[0107] Example 5: 2-(3,5-dichloro-4-(4-hydroxy-3-(trifluoromethyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 5) [ka]

[0108] Step 1: 4-(2,6-dichloro-4-nitrophenoxy)-2-(trifluoromethyl)phenol

[0109] To acetonitrile (50 mL), 2-trifluoromethylbenzene-1,4-diol (3.56 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-dichloro-2-fluoro-5-nitrobenzene (3.00 g, 14.3 mmol) were added. The mixture was heated to 48 °C and stirred at 48 °C for 8 hours. After completion of the reaction, the reaction solution was cooled to room temperature and concentrated under reduced pressure to remove acetonitrile. Water (50 mL) and ethyl acetate (50 mL) were added to the concentrate, and the pH was adjusted to 2 - 3 with 10% aqueous hydrochloric acid. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined. The organic phase was washed with saturated aqueous sodium chloride (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain a crude product of 4-(2,6-dichloro-4-nitrophenoxy)-2-(trifluoromethyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 - 15:1) to obtain 3.32 g of the product in a yield of 62%. MS (ESI) m / z: 369.1 [M + H] + .

[0110] Step 2 4-(4-Amino-2,6-dichlorophenoxy)-2-(trifluoromethyl)phenol

[0111] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dichloro-4-nitrophenoxy)-2-(trifluoromethyl)phenol (3.08 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.55 g of product in 90% yield. MS (ESI) m / z: 339.1 [M + H] + .

[0112] Step 3 (2-Cyano-2-(2-(3,5-Dichloro-4-(4-Hydroxy-3-Trifluoromethylphenoxy)phenyl)Hydrazinylidene)Acetyl)Carbamate Ethyl Ester Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dichlorophenoxy)-2-(trifluoromethyl)phenol (2.62 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. After addition, stirring was continued at 0-5°C for 30 minutes. A solution of pyridine (30 mL), water (40 mL), and ethyl (2-cyanoacetyl) carbamate (1.12 g, 7.74 mmol) was added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.93 g of product in 75% yield. MS (ESI) m / z: 506.3 [M + H] + .

[0113] Step 4: 2-(3,5-dichloro-4-(4-hydroxy-3-(trifluoromethyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0114] Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-trifluoromethylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.70 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.3 g of product in 50% yield. MS (ESI) m / z: 460.1 [M + H] + . 1 HNMR (400 MHz, DMSO-d6): δ 13.0 (br s, 1H), 9.70 (s, 1H), 7.80(s, 2H), 7.38 (d, , 1H), 7.26 (d, 1H), 6.84 (d, 1H). [Examples]

[0115] Example 6: 2-(3,5-dichloro-4-(4-hydroxy-3-(deuteromethyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 6) [ka]

[0116] Step 1: 4-(2,6-dichloro-4-nitrophenoxy)-2-(deuterium-methyl)phenol

[0117] 50 mL of acetonitrile was mixed with 2-deuteromethylbenzene-1,4-diol (2.55 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-dichloro-2-fluoro-5-nitrobenzene (3.00 g, 14.3 mmol). The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid aqueous solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride aqueous solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to dryness to obtain the crude product of 4-(2,6-dichloro-4-nitrophenoxy)-2-(deuterium methyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 to 15:1) to obtain 2.95 g in yield of 65%. MS (ESI) m / z: 318.1 [M + H] + .

[0118] Step 2: 4-(4-amino-2,6-dichlorophenoxy)-2-(deuterium-methyl)phenol

[0119] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dichloro-4-nitrophenoxy)-2-(deuterium-methyl)phenol (2.66 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.24 g of product in 93% yield. MS (ESI) m / z: 288.2 [M + H] + .

[0120] Step 3 (2-Cyano-2-(2-(3,5-Dichloro-4-(4-Hydroxy-3-Deuteromethylphenoxy)phenyl)Hydrazinylidene)Acetyl)Carbamate Ethyl Ester

[0121] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dichlorophenoxy)-2-(deuterium-methyl)phenol (2.22 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. After addition, stirring was continued at 0-5°C for 30 minutes. A solution of pyridine (30 mL), water (40 mL), and ethyl (2-cyanoacetyl) carbamate (1.12 g, 7.74 mmol) was added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.6 g of product in 75% yield. MS (ESI) m / z: 455.3 [M + H] + . Step 4: 2-(3,5-dichloro-4-(4-hydroxy-3-(deuterium-methyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-douteromethylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.70 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.04 g of product in 45% yield. MS(ESI) m / z: 409.2 [M + H] + . 1 H NMR(400MHz, DMSO-d6):δ 13.50(br s, 1H), 9.41(s, 1H), 7.84(s, 2H), 7.84(s, 2H).7.84 (s, 2H), 7.07 (d, 1H), 6.99 (s, 1H), 6.79 (d, 1H). [Examples]

[0122] Example 7: 2-(3,5-dimethyl-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 7) [ka]

[0123] Step 1: 4-(2,6-dimethyl-4-nitrophenoxy)-2-(isopropenyl)phenol

[0124] To 50 mL of acetonitrile, 3.00 g, 20.0 mmol of 2-isopropenylbenzene-1,4-diol, 7.57 g, 71.4 mmol of sodium carbonate, and 2.42 g, 14.3 mmol of 1,3-dimethyl-2-fluoro-5-nitrobenzene were added. The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid aqueous solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride aqueous solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to dryness to obtain the crude product of 4-(2,6-dimethyl4-nitrophenoxy)-2-(isopropenyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 to 15:1) to obtain 2.99 g of product in 69.8% yield. MS (ESI) m / z: 300.3 [M + H] + .

[0125] Step 2: 4-(4-amino-2,6-dimethylphenoxy)-2-(isopropenyl)phenol

[0126] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dimethyl-4-nitrophenoxy)-2-(isopropenyl)phenol (2.85 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.14 g of product in 95% yield. MS (ESI) m / z: 270.3 [M + H] + .

[0127] Step 3 (2-Cyano-2-(2-(3,5-Dimethyl-4-(4-Hydroxy-3-Isopropenylphenoxy)phenyl)Hydrazinylidene)Acetyl)Carbamate Ethyl Ester

[0128] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dimethylphenoxy)-2-(isopropenyl)phenol (2.08 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.53 g of product in 75% yield. MS (ESI) m / z: 437.5 [M + H] + .

[0129] Step 4: 2-(3,5-dimethyl-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0130] Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dimethyl-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.47 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.05 g of product in yield of 22%. MS(ESI) m / z: 391.4 [M + H] + . 1 H NMR(400MHz, DMSO-d6):δ 13.40(br s, 1H), 9.38(s, 1H), 7.50 (s, 2H), 7.16 (m, 2H), 6.91 (s, 1H), 5.15 (s, 2H), 2.30 (s, 3H), 2.15 (s, 6H). [Examples]

[0131] Example 8: 2-(3,5-dibromo-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 8) [ka]

[0132] Step 1: 4-(2,6-dibromo-4-nitrophenoxy)-2-(isopropenyl)phenol

[0133] 50 mL of acetonitrile was mixed with 2-isopropenylbenzene-1,4-diol (3.00 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-dibromo-2-fluoro-5-nitrobenzene (4.28 g, 14.3 mmol). The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain the crude product of 4-(2,6-dibromo-4-nitrophenoxy)-2-(isopropenyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1~15:1) to obtain 3.68 g of the product in 60% yield. MS (ESI) m / z: 430.1 [M + H] + .

[0134] Step 2: 4-(4-amino-2,6-dibromophenoxy)-2-(isopropenyl)phenol

[0135] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dibromo-4-nitrophenoxy)-2-(isopropenyl)phenol (3.59 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 3.18 g of product in 95% yield. MS (ESI) m / z: 400.1 [M + H] + .

[0136] Step 3 Ethyl (2-cyano-2-(2-(3,5-dibromo-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazinyl)acetyl)carbamate

[0137] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dibromophenoxy)-2-(isopropenyl)phenol (3.09 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 3.24 g of product in 74% yield. MS (ESI) m / z: 567.2 [M + H] + .

[0138] Step 4: 2-(3,5-dibromo-4-(4-hydroxy-3-(isopropenyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0139] Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dibromo-4-(4-hydroxy-3-isopropenylphenoxy)phenyl)hydrazinylidene)acetyl)carbamate ethyl ester (3.20 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.32 g of product in 45% yield. MS(ESI) m / z: 521.1 [M + H] + .1H NMR (400MHz, DMSO-d6):δ 13.35(br s, 1H), 9.45(s, 1H), 7.77(s, 2H)7.77 (s, 2H), 7.14 (m, 2H), 6.91 (s, 1H), 5.10 (s, 2H), 2.35 (s, 3H). [Examples]

[0140] Example 9: 2-(4-(4-hydroxy-3-(trifluoromethyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 9) [ka]

[0141] Step 1 (2,6-dimethyl-4-nitrophenyl)(4-methoxy-3-(trifluoromethyl)phenyl) methanol

[0142] Add 2-bromo-1,3-dimethyl-5-nitrobenzene (2.83 g, 12.3 mmol) to tetrahydrofuran (80 mL), and -70 ° The mixture was cooled to °C. n-Butylithium (10 mL, 24.5 mmol) was added dropwise to the above mixture and stirred for 20 minutes. 4-Methoxy-3-(trifluoromethyl)benzaldehyde (3.0 g, 14.7 mmol) was added dropwise to the above reaction solution. 4-Methoxy-3-(trifluoromethyl)benzaldehyde (3.0 g, 14.7 mmol) and tetrahydrofuran (20 mL) were added dropwise to the above reaction solution, and the mixture was cooled to -70°C. ° The mixture was stirred at °C for 60 minutes. After the reaction was complete, the reaction solution was poured into a saturated ammonium chloride aqueous solution (300 mL) and extracted with ethyl acetate (150 mL x 2). The organic phases were combined and washed with a saturated sodium chloride aqueous solution (200 mL), and anhydrous magnesium sulfate was dried. The mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 10-95% (%B)) to obtain 2.84 g of product in yield of 65%. MS (ESI) m / z: 336.3 [M + H] + .

[0143] Step 2: 2-(4-methoxy-3-(trifluoromethyl)benzyl)-1,3-dimethyl-5-nitrobenzene

[0144] (2,6-dimethyl-4-nitrophenyl)(4-methoxy-3-(trifluoromethyl)phenyl)methanol (2.75 g, 7.74 mmol) and trifluoroacetic acid (10 mL) were added to dichloromethane (40 mL). Triethylsilane (20 mL) was added dropwise to the mixture at room temperature, and the mixture was allowed to react overnight at room temperature. After the reaction was complete, the reaction solution was poured into saturated sodium bicarbonate aqueous solution and extracted with dichloromethane (300 mL x 2). The organic phases were combined, the organic phase was washed with saturated sodium chloride aqueous solution (250 mL), and dried over anhydrous magnesium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure until dry. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5-85% (% B)) to obtain the following: MS (ESI) m / z: 340.3 [M + H] + .

[0145] Step 3: 4-(2,6-dimethyl-4-nitrobenzyl)-2-(trifluoromethyl)phenol

[0146] 2-(4-methoxy-3-(trifluoromethyl)benzyl)-1,3-dimethyl-5-nitrobenzene (1.41 g, 4.15 mmol) and pyridine hydrochloride (17.0 g) were stirred at 160°C for 16 hours. After the reaction was complete, the temperature was lowered to room temperature, ethyl acetate (200 mL) and water (200 mL) were added, and the mixture was stirred for 30 minutes. The liquids were then partitioned, and the aqueous phase was extracted with ethyl acetate (100 mL). The organic phase was combined with the aqueous phase, washed with saturated sodium chloride aqueous solution (200 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure until dry. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5-75% (% B)) to obtain 945 mg of product in 70% yield. MS(ESI) m / z: 326.3 [M + H] + .

[0147] Step 4: 4-(4-amino-2,6-dimethylbenzyl)-2-(trifluoromethyl)phenol

[0148] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dimethyl-4-nitrobenzyl)-2-(trifluoromethyl)phenol (2.73 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated sodium bicarbonate aqueous solution (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.30 g of product in 93% yield. MS (ESI) m / z: 296.3 [M + H] + .

[0149] Step 5 (Ethyl 2-cyano-2-(2-(4-(4-hydroxy-3-(trifluoromethyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate

[0150] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dimethylbenzyl)-2-(trifluoromethyl)phenol (2.29 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.68 g of product in 75% yield. MS (ESI) m / z: 463.4 [M + H] + .

[0151] Step 6: 2-(4-(4-hydroxy-3-(trifluoromethyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0152] Glacial acetic acid (30 mL), (2-cyano-2-(2-(4-hydroxy-3-(trifluoromethyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.62 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.18 g of product in 50% yield. MS (ESI) m / z: 417.4 [M + H] + . 1 H NMR (400 MHz, DMSO-d6): δ 12.95 (br s, 1H), 9.50 (s, 1H), 7.42 (s, 2H), 7.32 (d, 1H), 7.08 (d, 1H), 6.76 (d1H), 3.96 (s, 2H), 2.18 (s, 6H). [Examples]

[0153] Example 10: 2-(4-(4-hydroxy-3-(deuterium-methyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 10) [ka]

[0154] Step 1 (2,6-dimethyl-4-nitrophenyl)(4-methoxy-3-(deuterium-methyl)phenyl) methanol

[0155] Add 2-bromo-1,3-dimethyl-5-nitrobenzene (2.83 g, 12.3 mmol) to tetrahydrofuran (80 mL), and -70 ° The mixture was cooled to °C. n-Butylithium (10 mL, 24.5 mmol) was added dropwise to the above mixture, and the mixture was kept warm and stirred for 20 minutes. 4-Methoxy-3-(deuteromethyl)benzaldehyde (2.25 g, 14.7 mmol) was added dropwise to the above reaction solution. 4-Methoxy-3-(deuteromethyl)benzaldehyde (2.25 g, 14.7 mmol) and tetrahydrofuran (20 mL) were added dropwise to the above reaction solution, and the mixture was cooled to -70°C. ° The mixture was stirred at °C for 60 minutes. After the reaction was complete, the reaction solution was poured into a saturated ammonium chloride aqueous solution (300 mL) and extracted with ethyl acetate (150 mL x 2). The organic phases were combined, washed with a saturated sodium chloride aqueous solution (200 mL), and dried over anhydrous magnesium sulfate. The mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The concentrate was purified by reverse-phase preparative liquid-phase separation (mobile phase A: water, mobile phase B: acetonitrile, gradient: 10-95% (%B)) to obtain 2.55 g of product in yield 68%. MS(ESI) m / z: 305.4 [M + H] + .

[0156] Step 2: 2-(4-methoxy-3-(deuterium-methyl)benzyl)-1,3-dimethyl-5-nitrobenzene

[0157] (2,6-dimethyl-4-nitrophenyl)(4-methoxy-3-(deuterium-methyl)phenyl)methanol (2.36 g, 7.74 mmol) and trifluoroacetic acid (10 mL) were added to dichloromethane (40 mL). Triethylsilane (20 mL) was added dropwise to the mixture at room temperature, and the mixture was allowed to react overnight at room temperature. After the reaction was complete, the reaction solution was poured into saturated sodium bicarbonate aqueous solution and extracted with dichloromethane (300 mL x 2). The organic phases were combined, the organic phase was washed with saturated sodium chloride aqueous solution (250 mL), and dried over anhydrous magnesium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure until dry. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5-85% (% B)) to obtain the following: MS (ESI) m / z: 289.4 [M + H]+ .

[0158] Step 3: 4-(2,6-dimethyl-4-nitrobenzyl)-2-(deuterium-methyl)phenol

[0159] 2-(4-methoxy-3-(deuterium-methyl)benzyl)-1,3-dimethyl-5-nitrobenzene (1.20 g, 4.15 mmol) and pyridine hydrochloride (17.0 g) were stirred at 160 °C for 16 hours. After the reaction was complete, the temperature was lowered to room temperature, ethyl acetate (200 mL) and water (200 mL) were added, and the mixture was stirred for 30 minutes. The liquids were then partitioned, and the aqueous phase was extracted with ethyl acetate (100 mL). The organic phase was combined with the aqueous phase, washed with saturated sodium chloride aqueous solution (200 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure until dry. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5-75% (% B)) to obtain 910 mg of product in 80% yield. MS(ESI) m / z: 275.3 [M + H] + .

[0160] Step 4: 4-(4-amino-2,6-dimethylbenzyl)-2-(deuterium-methyl)phenol

[0161] In a reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dimethyl-4-nitrobenzyl)-2-(deuteromethyl)phenol (2.3 g, 8.38 mmol), aqueous solution of ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and powdered iron (2.82 g, 50.3 mmol) were added. The mixture was heated to 70°C and stirred at 70°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. Water (50 mL), saturated aqueous solution of sodium bicarbonate (30 mL), and ethyl acetate (50 mL) were added to the reaction mixture, and after stirring for 10 minutes, the mixture was filtered. The filtrate was extracted with ethyl acetate (20 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride (50 mL), dried on anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 1.95 g of product in 95% yield. MS (ESI) m / z: 245.3 [M + H] + .

[0162] Step 5 (Ethyl 2-cyano-2-(2-(4-(4-hydroxy-3-(deuterium-methyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate

[0163] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dimethylbenzyl)-2-(deuterium methyl)phenol (1.90 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.42 g of product in 76% yield. MS (ESI) m / z: 412.5 [M + H] + .

[0164] Step 6: 2-(4-(4-hydroxy-3-(deuterium-methyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0165] Glacial acetic acid (30 mL), (2-cyano-2-(2-(4-hydroxy-3-(deuterium-methyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.33 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.1 g of product in yield 53%. MS(ESI) m / z: 366.4 [M + H] + . 1 H NMR(400MHz, DMSO-d6):δ 12.87(br s, 1H), 9.26(s, 1H), 7.42 (s, 2H), 6.93 (d, 1H), 6.87 (d, 1H), 6.71 (d, 1H), 3.96 (s, 2H), 2.10 (s, 6H). [Examples]

[0166] Example 11: 2-(3,5-dichloro-4-(4-hydroxy-3-(allyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 11) [ka]

[0167] Step 1: 4-(2,6-dichloro-4-nitrophenoxy)-2-(allyl)phenol

[0168] 50 mL of acetonitrile was mixed with 2-allylbenzene-1,4-diol (3.00 g, 20.0 mmol), sodium carbonate (7.57 g, 71.4 mmol), and 1,3-dichloro-2-fluoro-5-nitrobenzene (3.00 g, 14.3 mmol). The mixture was heated to 48°C and stirred at 48°C for 8 hours. After the reaction was complete, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the acetonitrile. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, and the pH was adjusted to 2-3 with 10% hydrochloric acid solution. The mixture was extracted with ethyl acetate (30 mL x 3) to combine the organic phases. The organic phases were washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to dryness to obtain the crude product of 4-(2,6-dichloro-4-nitrophenoxy)-2-(allyl)phenol. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1~15:1) to obtain 2.85 g of product in 59% yield. MS(ESI)m / z: 340.0 [M+H] + .

[0169] Step 2: 4-(4-amino-2,6-dichlorophenoxy)-2-(allyl)phenol

[0170] To the reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dichloro-4-nitrophenoxy)-2-(allyl)phenol (2.85 g, 8.38 mmol), aqueous ammonium chloride (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70 °C and stirred at 70 °C for 3 hours. After completion of the reaction, the reaction solution was cooled to room temperature. Water (50 mL), saturated aqueous sodium hydrogen carbonate (30 mL) and ethyl acetate (50 mL) were added to the reaction solution, stirred for 10 minutes, and then filtered. The filtrate was extracted with ethyl acetate (20 mL × 3), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.42 g of the product in a yield of 93%. MS(ESI) m / z: 310.1 [M+H] + .

[0171] Step 3: Ethyl (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-allylphenoxy)phenyl)hydrazinyl)acetyl)carbamate

[0172] To the reaction flask, anhydrous ethanol (30 mL), 6M aqueous hydrochloric acid (40 mL), 4-(4-amino-2,6-dichlorophenoxy)-2-(allyl)phenol (2.40 g, 7.74 mmol) were added. The mixture was cooled to 5 °C, and an aqueous solution (3 mL) of sodium nitrite (534 mg, 7.74 mmol) was added dropwise to the mixture. A solution of pyridine (30 mL), ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) was added dropwise to the reaction solution, and stirred at 5 °C for 2 hours. Filtration was carried out, and the filter cake was washed with water (10 mL × 3). The filter cake was dried under reduced pressure to obtain 2.7 g of the product in a yield of 73%. MS(ESI) m / z: 477.1 [M+H] + .

[0173] Step 4: 2-(3,5-dichloro-4-(4-hydroxy-3-(allyl)phenoxy)phenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile Glacial acetic acid (30 mL), (2-cyano-2-(2-(3,5-dichloro-4-(4-hydroxy-3-allylphenoxy)phenyl)hydrazone)acetyl)carbamate ethyl ester (2.70 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% aqueous ammonium bicarbonate = 20%~80%) to obtain 1.05 g of product in 43% yield. MS(ESI) m / z: 431.0 [MH] - . [Examples]

[0174] Example 12: 2-(4-(4-hydroxy-3-(allyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile (Compound 12) [ka]

[0175] Step 1: (2,6-dimethyl-4-nitrophenyl)(4-allyloxyphenyl) methanol

[0176] Add 2-bromo-1,3-dimethyl-5-nitrobenzene (2.83 g, 12.3 mmol) to tetrahydrofuran (80 mL) and -70 ° The mixture was cooled to °C. n-Butylithium (10 mL, 24.5 mmol) was added dropwise to the mixture and stirred at a constant temperature for 20 minutes. 4-Allyloxybenzaldehyde (3.0 g, 14.7 mmol) and tetrahydrofuran (20 mL) were added dropwise to the reaction solution and stirred at -70°C. °The mixture was stirred at °C for 60 minutes. After the reaction was complete, the reaction solution was poured into a saturated ammonium chloride aqueous solution (300 mL) and extracted with ethyl acetate (150 mL x 2). The organic phases were combined, washed with a saturated sodium chloride aqueous solution (200 mL), and dried over anhydrous magnesium sulfate. The mixture was filtered, the filtrate was concentrated to dryness under reduced pressure, and the concentrate was purified by reverse-phase preparative liquid-phase extraction. After reverse-phase preparative liquid-phase extraction (mobile phase A: water, mobile phase B: acetonitrile, gradient: 10-95% (%B)), the product was 2.84 g, with a yield of 65%. MS (ESI) m / z: 314.1 [M + H] + .

[0177] Step 2: 2-(4-allyloxy)benzyl-1,3-dimethyl-5-nitrobenzene

[0178] Dichloromethane (40 mL) was mixed with (2,6-dimethyl-4-nitrophenyl)(4-allyloxyphenyl)methanol (2.75 g, 7.74 mmol) and trifluoroacetic acid (10 mL). Triethylsilane (20 mL) was added dropwise to the mixture at room temperature, and the mixture was allowed to react overnight at room temperature. After the reaction was complete, the reaction solution was poured into saturated sodium bicarbonate aqueous solution and extracted with dichloromethane (300 mL x 2). The organic phases were combined, the organic phase was washed with saturated sodium chloride aqueous solution (250 mL), and dried over anhydrous magnesium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure until dry. The concentrate was purified by reverse-phase preparative liquid-phase purification (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5-85% (%B)) to obtain the following: MS (ESI) m / z: 298.2 [M + H] + .

[0179] Step 3: 4-(2,6-dimethyl-4-nitrobenzyl)-2-(allyl)phenol

[0180] 2-(4-allyloxy)benzyl-1,3-dimethyl-5-nitrobenzene (1.41 g, 4.15 mmol) was dissolved in dichloromethane (20 mL), diethylaluminum chloride (1.2 eq) was added, and the mixture was stirred at room temperature for 3 hours. Then, water (5 mL) was added to quench the reaction, and the mixture was stirred for 30 minutes. The organic phase was washed with saturated aqueous sodium chloride (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The concentrate was purified by reverse-phase preparative liquid chromatography (mobile phase A: water, mobile phase B: acetonitrile, gradient: 5 - 75% (%B)) to obtain 945 mg of the product with a yield of 70%. MS(ESI) m / z: 298.2 [M + H] + .

[0181] Step 4 4-(4-amino-2,6-dimethylbenzyl)-2-(allyl)phenol

[0182] Into the reaction flask, tetrahydrofuran (20 mL), anhydrous methanol (20 mL), 4-(2,6-dimethyl-4-nitrobenzyl)-2-(trifluoromethyl)phenol (2.73 g, 8.38 mmol), aqueous ammonium chloride solution (4.48 g, 83.8 mmol) (20 mL), and iron powder (2.82 g, 50.3 mmol) were added. The mixture was heated to 70 °C and stirred at 70 °C for 3 hours. After the reaction was completed, the reaction solution was cooled to room temperature. Water (50 mL), saturated aqueous sodium hydrogen carbonate solution (30 mL) and ethyl acetate (50 mL) were added to the reaction solution, and the mixture was stirred for 1 minute and then filtered. The filtrate was extracted with ethyl acetate (20 mL × 3), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated to dryness under reduced pressure to obtain 2.30 g of the product with a yield of 93%. MS (ESI) m / z: 268.2 [M + H] + .

[0183] Step 5 (ethyl 2-cyano-2-(2-(4-(4-hydroxy-3-(allyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate

[0184] Anhydrous ethanol (30 mL), 6M hydrochloric acid aqueous solution (40 mL), and 4-(4-amino-2,6-dimethylbenzyl)-2-(allyl)phenol (2.29 g, 7.74 mmol) were added to a reaction flask. The mixture was cooled to 5°C, and an aqueous solution of sodium nitrite (534 mg, 7.74 mmol) (3 mL) was added dropwise to the mixture. Pyridine (30 mL) and a solution of ethyl cyanoacetylcarbamate (1.12 g, 7.74 mmol) in water (40 mL) were added dropwise to the reaction mixture, and the mixture was stirred at 5°C for 2 hours. The mixture was filtered, and the filter cake was washed with water (10 mL x 3). The filter cake was dried under reduced pressure to obtain 2.68 g of product in 75% yield. MS (ESI) m / z: 435.2 [M + H] + .

[0185] Step 6: 2-(4-(4-hydroxy-3-(allyl)benzyl)-3,5-dimethylphenyl)-3,5-dioxo-2,3,4,5-tetrahydro-1,2,4-triazine-6-carbonitrile

[0186] Glacial acetic acid (30 mL), (2-cyano-2-(2-(4-hydroxy-3-(allyl)benzyl)-3,5-dimethylphenyl)hydrazinylidene)acetyl)carbamate ethyl ester (2.62 g, 5.66 mmol), and sodium acetate (928 mg, 11.3 mmol) were added to the reaction flask. The mixture was heated to 118°C and stirred at 118°C for 3 hours. The reaction mixture was cooled to 60°C and concentrated to dryness under reduced pressure. The concentrate was purified by preparative liquid-phase purification using a C18 column (mobile phase: acetonitrile: 0.1% ammonium bicarbonate aqueous solution = 20%~80%) to obtain 1.18 g of product in 50% yield. MS(ESI) m / z: 389.2 [M + H] + .

[0187] Examples of effects

[0188] Example 1 of the effect: TR-FRET (time-resolved fluorescence resonance energy transfer) thyroid receptor co-activation assay

[0189] Experimental method: (1) 1 M DTT (disulfide) was added to TR-FRET Coregulator Buffer C (purchased from Thermofisher). A complete TR-FRET Coregulator Buffer C with a final concentration of 5 mM DTT was prepared using Threitol. (2) Add 100 nL of the compound to be tested per well at a concentration of 200 X. For the control, add 100 nL of DMSO. (3) Add 10 μL of TR-FRET Coregulator Buffer C to each well. (4) Prepare the 4X TR-LBD (thyroid hormone ligand binding domain) using the pre-cooled Complete TR-fret Coregulator Buffer C. (5) Add 5 μL of 4X TR-LBD to the experimental plate and prepare a 0.4 μM luciferase-SRC2-2 (purchased from Thermofisher) (4x) and an 8 nM Tb anti-GST (purchased from Thermofisher) (4x) solution. Add 5 μL of 4X peptide / 4X antibody solution (purchased from Thermofisher) to the experimental plate. (6) The 384-well plate was gently mixed in a plate shaker and incubated at room temperature for 2 hours, away from light. The wavelengths were set to 520 nm and 495 nm according to the instrument settings, and the specific parameters were set as follows: [Table 1]

[0190] Target compounds: Thyroxine T3 (positive control), MGL3196 (positive compound), and compounds of embodiments of the present invention. result [Table 2]

[0191] As a result, compounds 1 and 3 of the present invention exhibited agonist activity against both TRα and TRβ, and their activity was already close to that of the positive control T3. The agonist effect against TRβ reached more than 90% of the agonist effect of T3. Furthermore, both compounds showed significantly higher EC50 values ​​and agonist activity against TRα and TRβ than compound MGL3196.

[0192] Example 2 of the effect: Functional testing of luciferasein using HEK293 / TRβ-luc cells

[0193] (1) Add 125 nL of HEK293 / TRβ-luc cells to each well of the assay plate to prepare a composite plate. Measure the cell density by counting, dilute the cell suspension with complete medium, and if the cell density is 4 × 10⁶, 5 It was adjusted to be / mL. (2) Dispense 25 μL of cells per well into an assay plate containing the compound to be tested, and incubate at 37°C in 5% CO2 for 24 hours. (3) 25 uL of Steady-Glo (Promega) was added to each well. (4) Centrifuge at 2000 RPM for 2 minutes to remove air bubbles. (5) Incubate at room temperature for 10 minutes and read the plate with Envision (Envision model 2105, PerkinElmer).

[0194] Target compounds: Thyroxine T3 (positive control), MGL3196 (positive control), and the target compound.

[0195] result [Table 3]

[0196] As a result, compound 1 in the present invention exhibited agonist activity against both TRα and TRβ, had a significantly higher EC50 value than compound MGL3196, and its selectivity for TRβ cells over TRα cells was 22.2 times higher, which was also significantly higher than that of MGL3196.

[0197] Example 3 of the effect: Efficacy and safety testing in an animal model of NASH

[0198] When C57BL / 6J male mice were continuously fed a CDAA-HFD diet (choline-deficient levulinic acid amino acid high-fat diet) from 6 weeks of age, a model was successfully created after 6 weeks (42 days). In this model, hepatic triglyceride secretion via VLDL (very low-density lipoprotein) was impaired, and the mice developed elevated serum ALT and AST levels, steatosis and inflammation within 3 weeks, and hepatic fibrosis within 5-6 weeks. The CDAA-HFD diet induces hepatic fat deposition and hepatic fibrosis in a short period without the characteristics of metabolic syndrome such as obesity, hyperglycemia, and hypertriglyceridemia, making it extremely useful for NASH disease research projects. In this study, we used this CDAA-HFD model to simulate the pathological processes and physiological states of NASH and to verify the therapeutic effects of compounds on early to mid-stage NASH.

[0199] Specific implementation program: [Table 4] Note: PO is administered orally (enteral administration), qd is administered once daily.

[0200] The groups were divided starting from day 42, when the model was successfully established, and administered according to the 42-day treatment regimen.

[0201] Drug preparation: Each dose group weighed an appropriate amount of sample powder, placed it in a 5 mL centrifuge tube, added an appropriate amount of 0.5% MC, mixed it using vortex shock, and prepared a solution of the corresponding concentration for use.

[0202] The animals' condition was observed and recorded after each administration. In case of death, a gross autopsy was performed, and the internal organs were visually inspected and recorded for any abnormalities. The animals' body weight was measured twice a week during the experiment. Figure 1 shows the body weight curves of animals administered for 42 days.

[0203] 2. Pathological Score: After measuring the body weight of all mice at appropriate time points, they were euthanized with high concentrations of carbon dioxide. Blood was collected by cardiac puncture, and the plasma was centrifuged at 7000 rpm for 10 minutes. Immediately afterward, it was placed on dry ice and stored at -80°C. Subsequently, blood biochemical indicators, including TCHO (total cholesterol) and LDL (low-density lipoprotein), were examined.

[0204] Liver tissue was harvested, weighed, and a portion of the liver (the same area in each animal) was excised. This tissue was fixed with 4% paraformaldehyde and subjected to histopathological analysis (HE staining and Sirius Red staining): the degree of hepatic fat and inflammatory cell infiltration, fibrosis, and NAS score (for the NAS scoring system, refer to the Chinese Medical Association's "Guidelines for the Diagnosis and Treatment of Non-Alcoholic Fatty Liver Disease" (revised edition, 2010). (Ishak scoring criteria, Journal of Hepatology 47 (2007) 598-607, Grading and staging systems for inflammation and fibrosis in chronic liver diseases.)).

[0205] 3.Statistical analysis Data are represented as mean + SEM. Statistical analysis of differences between groups was performed using SPSS statistical software, specifically one-way analysis of variance (ANOVA) followed by Dunnett's test. A p-value of less than 0.05 indicates statistical significance between the data of the two groups. In the graphs of this example, * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001.

[0206] 4. Experimental Results

[0207] 4.1 Lipid-lowering effect of Compound 1 Figure 2 shows TCHO (total cholesterol), and Figure 3 shows LDL (low-density lipoprotein). Compound 1 showed a significant reduction compared to the normal diet group, the model group, and the fenofibrate group, confirming the liver fat-reducing effect of Compound 1 as a THR-β inhibitor.

[0208] 4.2 Hepatocyte balloon formation and inflammation score of Compound 1 Hepatocyte balloon formation can indicate the severity of hepatic fat accumulation. As shown in Figure 4, compound 1 significantly reduced the hepatocyte balloon score. Another key indicator of NASH is the degree of hepatitis cell infiltration. As shown in Figure 5, compound 1 showed good suppression and reversal of hepatitis, suggesting the potential of compound 1 to treat NASH.

[0209] 4.3 Fibrosis of Compound 1 and NAS score The liver fibrosis score is shown in Figure 6, and the NAS score is shown in Figure 7. Compound 1 significantly improved both the liver NAS score and fibrosis. When the lipid-lowering effect of compound 2 was tested using the same method as above, compound 2 showed a significant reduction compared to the normal diet group, the model group, and the fenofibrate group, confirming that compound 2 effectively reduces liver fat as a THR-β inhibitor. When the hepatocyte balloon-like changes and inflammation scores of compound 2 were tested using the same method as above, compound 2 also showed good hepatitis suppression and recovery effects.

[0210] Example of effect 4: Maximum load tolerance test

[0211] Twenty-four C57BL / 6J mice (male and female) were orally administered compound 1 as a single dose at doses of 30, 100, and 300 mg / kg in a 0.1% tween80 + 0.5% MC aqueous solution. Clinical symptoms and body weight were recorded once daily for three days after administration. On the fourth day, the mice were euthanized, and blood was collected for hematological and biochemical tests.

[0212] Mice tolerated compound 1 well, and no significant abnormalities in body weight or clinical signs were observed during the experiment. Hematological and blood biochemistry results were also normal. The maximum tolerated dose (MTD) was ≥300 mg / kg, suggesting a favorable safety profile for compound 1.

[0213] Example of effect 5: hERG test

[0214] The inhibitory effect of compound 1 on hERG potassium current in CHO cells was tested using fully automated membrane clamp QPatch technology.

[0215] Specific implementation program:

[0216] 1. Cell preparation CHO-hERG cells 175cm 2 The cells were cultured in a culture flask, and when the cell density reached 60-80%, the culture medium was removed and the cells were removed. After washing once with mL of PBS (phosphate-buffered saline), digestion was carried out with 3 mL of Detachin. Once digestion was complete, neutralization was performed by adding 7 mL of culture medium, the cells were centrifuged, the supernatant was aspirated, and the cells were resuspended by adding 5 mL of culture medium until the cell density was 2–5 × 10⁶ / mL.

[0217] 2. Electrophysiological Recording Process The single-cell high-impedance sealing and whole-cell patterning processes were all automated using a Qpatch device. After obtaining the whole-cell recording pattern, the cells were clamped at -80mV, a preconditioning voltage of -50mV was applied for 50ms, followed by a depolarizing stimulus of +40mV for 5 seconds, maintaining repolarization to -50mV for 5 seconds, and then returning to -80mV. This voltage stimulus was applied every 15 seconds, recorded for 2 minutes, then added to extracellular fluid and recorded for 5 minutes before initiating the administration process. The highest concentration of compound 1 tested was 40.00 μM, with six other concentrations being tested in the following order: 40.00, 13.33, 4.44, 1.48, 0.49, and 0.16 μM. The final test concentrations did not contain more than 0.2% DMSO, and this concentration of DMSO did not affect hERG potassium channels. The compound concentration was administered starting from the lowest test concentration, for 2.5 minutes at each concentration, and after administering all concentrations consecutively, the positive control compound cisapride was administered. A minimum of 3 cells (n≧3) were tested at each concentration.

[0218] 3. Data Analysis The experimental data was analyzed using GraphPad Prism 5.0 software.

[0219] 4. Experimental Results [Table 5] Note: Cells treated with compound 1 were unable to maintain normal sealing at a concentration of 40 μM. Data at this concentration are not included in the statistics.

[0220] The results showed that compound 1 had little effect on hERG potassium current in CHO cells, suggesting that there are few safety concerns for the heart.

Claims

1. The compound represented by formula I, or a pharmaceutically acceptable salt thereof, characterized by the following structure: 【Chemistry 1】 Here, A is O or CH 2 It is; M is 【Chemistry 2】 It is; X and Y are, independently, chlorine, bromine, iodine, and isotope I. 124 I or 131 I, or C 1 ~C 6 It is alkyl; R 1 is hydrogen, C 1 to C 6 alkyl, C 2 to C 6 alkenyl, one or more fluoro-substituted C 1 to C 6 alkyl, one or more fluoro-substituted C 2 to C 6 alkenyl, one or more deuterium-substituted C 1 to C 6 alkyl, or one or more deuterium-substituted C 2 to C 6 alkenyl; The compound represented by formula I is 【Transformation 3】 isn't it.

2. X and Y are independently chlorine, bromine, iodine, or CH 3 It is; and / or, R 1 However, C 2 ~C 6 Alkenyl or one or more fluorosubstituted C 1 ~C 6 A compound represented by formula I as described in claim 1, which is alkyl, or a pharmaceutically acceptable salt thereof.

3. R 1 C 2 ~C 6 When it is an alkenyl group, the C 2 ~C 6 Alkenyl group 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】 or 【Chemistry 11】 It is; and / or, R 1 C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl compounds include methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and / or, R 1 one or more fluorosubstituted C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl compounds include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and / or, R 1 C with one or more deuterium substitutions 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and “one or more” means one or three; and / or, R 1 one or more fluorosubstituted C 2 ~C 6 If it is an alkenyl, then C 2 ~C 6 Alkenil is 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 [Chemistry 18] or 【Chemistry 19】 It is; and / or, R 1 C with one or more deuterium substitutions 2 ~C 6 If it is an alkenyl, then C 2 ~C 6 Alkenil is 【Chemistry 20】 【Chemistry 21】 【Chemistry 22】 【Chemistry 23】 【Chemistry 24】 【Chemistry 25】 【Chemistry 26】 or 【Chemistry 27】 The aforementioned "one or more" means one or three; and / or X is C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl compounds include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl; and / or Y is C 1 ~C 6 If it is alkyl, then C 1 ~C 6 The compound represented by formula I according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the alkyl is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl.

4. R 1 C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl is isopropyl; and / or, R 1 one or more fluorosubstituted C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl is, 【Chemistry 28】 or 【Chemistry 29】 It is; and / or, R 1 C with one or more deuterium substitutions 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl is, 【Transformation 30】 It is; and / or X is C 1 ~C 6 If it is alkyl, then C 1 ~C 6 Alkyl is methyl; and / or Y is C 1 ~C 6 If it is alkyl, then C 1 ~C 6 The alkyl is methyl, wherein the compound represented by formula I according to claim 3 or a pharmaceutically acceptable salt thereof.

5. A is O or CH 2 It is; X and Y are, independently, chlorine, bromine, iodine, or C 1 ~C 6 It is alkyl; M is 【Chemistry 31】 and; R 1 is hydrogen, C 1 to C 6 alkyl, C 2 to C 6 alkenyl, one or more fluorine-substituted C 1 to C 6 alkyl, one or more fluorine-substituted C 2 to C 6 alkenyl, one or more deuterium-substituted C 1 to C 6 alkyl, or one or more deuterium-substituted C 2 to C 6 alkenyl, and is a compound represented by formula I according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

6. A is O; X and Y are independently chlorine, bromine, or iodine; M is 【Chemistry 32】 And; R 1 C 2 ~C 6 Alkenyl or one or more fluorosubstituted C 1 ~C 6 A compound represented by formula I according to any one of claims 1 to 3, which is alkyl, or a pharmaceutically acceptable salt thereof.

7. A is O; X and Y are independently chlorine or bromine; M is 【Transformation 33】 And; R 1 C 2 ~C 6 Alkenyl, or C substituted with one fluorine atom 1 ~C 6 A compound represented by formula I according to any one of claims 1 to 3, which is alkyl, or a pharmaceutically acceptable salt thereof.

8. In the formula, R 1 teeth 【Transformation 34】 【Chemistry 35】 【Transformation 36】 【Chemistry 37】 or 【Transformation 38】 It is; and / or 【Chemistry 39】 teeth, 【Chemistry 40】 【Chemistry 41】 【Chemistry 42】 【Chemistry 43】 【Chemistry 44】 【Chemistry 45】 【Chemistry 46】 【Chemistry 47】 or 【Chemistry 48】 The compound represented by formula I as described in claim 5, or a pharmaceutically acceptable salt thereof.

9. The compound represented by formula I is any of the following, the compound represented by formula I according to claim 5 or a pharmaceutically acceptable salt thereof: 【Chemistry 49】 [Transformation 50] 【Chemistry 51】 【Chemistry 52】 【Chemistry 53】 【Chemistry 54】 【Transformation 55】 【Transformation 56】 or 【Chemistry 57】

10. A method for preparing a compound represented by formula I, comprising the step of subjecting a compound represented by formula II-a to the following ring-closing reaction in a solvent in the presence of a base to obtain a compound represented by formula I, wherein M, X, Y, and A are defined as described in any of claims 1 to 5, and R 6 is C 1 ~C 6 Methods of being alkyl; 【Chemistry 58】 。

11. M, A, X, and Y are defined as described in any one of claims 1 to 5; and R 6 A compound represented by formula II-a, defined as described in claim 10. 【Chemistry 59】

12. The compound represented by formula II-a according to claim 11, wherein the compound represented by formula II-a is any of the following compounds. 【Transformation 60】 【Chemistry 61】 【Transformation 62】 【Transformation 63】 【Chemistry 64】 【Transformation 65】 【Chemical Formula 66】 【Transformation 67】 or 【Transformation 68】 。

13. A pharmaceutical composition characterized by containing substance A and one or more pharmaceutically acceptable carriers, A pharmaceutical composition wherein substance A is a compound represented by formula I as described in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof.

14. The use of substance B in the preparation of THR-β agonists, The use wherein substance B is a compound represented by formula I as described in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 13.

15. The use of substance B in the preparation of a medicine for the treatment and / or prevention of diseases related to THR-β, The use wherein substance B is a compound represented by formula I as described in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 13.

16. The use of substance B in the preparation of a medicine for the treatment and / or prevention of diseases related to THR-β, The substance B is a compound represented by formula I as described in any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 13. Use if the aforementioned disease is one or more of the following: non-alcoholic fatty liver disease, obesity, hepatic fibrosis, type 2 diabetes, and primary hypercholesterolemia.

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