Alkoxy thiophene acyl arylamine compounds and use thereof

Abstract

The present invention provides alkoxy thiophene acyl arylamine compounds and a use thereof, comprising alkoxy thiophene acyl arylamine compounds having a structure as shown in formula (1), or pharmaceutically acceptable salts thereof. The alkoxy thiophene acyl arylamine compounds provided by the present invention are a class of novel sphingomyelin synthase inhibitors, which have significant inhibitory activity against SMS2, ideal physical and chemical properties such as stability and water solubility, and low potential for toxic side effects, and can be further made into a drug for preventing or treating diseases related to abnormal sphingomyelin levels.

Description

Alkoxythiophene carboxylic acid aromatic amide compounds and their applications Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and more particularly relates to sphingomyelin synthase inhibitor alkoxythiophene carboxylic acid aromatic amide compounds and applications thereof. Background Art

[0002] Atherosclerosis (AS) is one of the main pathological bases of many cardiovascular and cerebrovascular diseases. Therefore, the research on anti-atherosclerotic drugs has become a hot topic in the current field of drug research and development.

[0003] Studies have proposed a variety of potential anti-atherosclerotic drug targets, including sphingomyelin synthase inhibitors, peroxisome proliferator-activated receptor (PPAR) agonists, apolipoprotein infusions, liver X receptor activators, and phospholipid transfer protein (PLTP) inhibitors. In particular, sphingomyelin (SM) and its metabolic enzymes mediate a range of cellular processes while inducing lipoprotein changes, suggesting a crucial role in the development and progression of atherosclerosis. It is believed that reducing plasma sphingomyelin levels or inhibiting SM synthesis could slow or prevent the development and progression of atherosclerosis.

[0004] Furthermore, researchers have discovered that sphingomyelin synthase (SMS) regulates the synthesis of ceramide (SM) from phosphatidylcholine (PC) and is a key enzyme in the final step of the de novo sphingomyelin synthesis pathway. There are three main isoforms of sphingomyelin synthase: sphingomyelin synthase 1 (SMS1), sphingomyelin synthase 2 (SMS2), and sphingomyelin synthase-related protein (SMSr). Studies have shown that inhibiting SMS2 to reduce sphingomyelin levels could be a new therapeutic approach for atherosclerosis. SMS2 has potential as a novel anti-atherosclerosis target, and SMS2-selective inhibitors have the potential to become novel anti-atherosclerotic drugs. Furthermore, studies have found that SMS2 deficiency protects against high-fat diet-induced obesity and insulin resistance. Furthermore, large, mature fatty plaques are rarely observed in the livers of SMS2-knockout mice, suggesting that SMS2 is involved in the formation of hepatic fatty plaques and can induce obesity and type 2 diabetes (Mitsutake et al., 2011). Small molecule inhibitors of sphingomyelin synthase have the potential to prevent and treat metabolic syndromes such as type 2 diabetes, obesity, and fatty liver disease. Literature reports that SMS2 gene knockout significantly improves inflammation, insulin resistance, and other metabolic syndromes in mice fed a high-fat diet (Mitsutake et al., 2011). Small molecule inhibitors of SMS2 have the potential to prevent and treat inflammatory diseases such as colitis and its associated diseases, colon cancer, pulmonary edema, and lung injury. Studies have shown that SMS2 promotes the epithelial-mesenchymal transition (EMT) in breast cancer cells (Zheng et al., 2019), drug resistance in colon cancer cells (Jin et al., 2016), growth and metastasis of ovarian cancer (Jing et al., 2021), M2 polarization of macrophages in the breast cancer tumor microenvironment (Deng et al., 2020), infiltration of malignant lymphomas (Taniguchi et al., 2020), and growth of leukemia cells (JP2021-165292A; WO2005032492). Small molecule inhibitors of sphingomyelin synthase have the potential to prevent and treat malignancies such as triple-negative breast cancer, colon cancer, leukemia, and ovarian cancer. SMS2 deficiency can inhibit the content and aggregation of sphingomyelin in the brain, reducing Aβ levels (Kidani et al., 2012; Yuyama et al., 2014; Hsiao et al., 2013). Small molecule inhibitors of SMS2 have the potential to prevent and treat central nervous system diseases, such as Alzheimer's disease and other neurodegenerative diseases.Literature has revealed that there are some lipid abnormalities in the tears of dry eye, including sphingomyelin (Ham et al., 2006; Paranjpe et al., 2019), suggesting that SMS2 inhibitors can be used to prevent and alleviate dry eye and other diseases related to abnormal sphingomyelin metabolism.

[0005] As research into the biological functions of SMS has deepened, SMS2 has been implicated in the development and progression of various diseases. In recent years, numerous SMS inhibitors have been discovered, some of which have been applied to the evaluation of therapeutic models for certain diseases. Aimin Meng et al. treated U937 human monocytic leukemia cells with D609, a selective tumor cytotoxic agent, and measured intracellular ceramide and DAG levels to assess the degree of SMS inhibition. They found that D609-induced cell death was time- and concentration-dependent, accompanied by high intracellular ceramide levels and low intracellular SM and DAG levels. Furthermore, treatment of cells with PKC inhibitors and agonists combined with D609 revealed that PKC inhibitors increased apoptosis, while agonists had the opposite effect, demonstrating that SMS is a potential target of D609 (Meng et al., 2004). Salma et al. found that Jaspine B, isolated from a sponge, significantly inhibited SMS activity in melanoma cells and induced apoptosis (Salma et al., 2009). Mahadeva MMSwamy et al. designed and synthesized a series of ceramide analogs, such as compound 17j, which exhibited certain SMS inhibitory effects (Swamy et al., 2018). Muhamad Aqmal Othman et al. discovered that Malabaricone C, a compound isolated from the fruit of Myristica cinnamomea King, inhibited SMS2 with low selectivity for SMS2 and SMS1, significantly reducing weight gain, glucose intolerance, and hepatic steatosis in HFD-induced mice (Swamy et al., 2018). Ryutaro Adachi et al. established a radioactive hydrogen substrate-labeled SMS activity assay system and, through high-throughput screening and structural optimization, identified a class of quinolone compounds (e.g., compound 3) with excellent SMS2 inhibitory activity and selectivity (Adachi et al., 2017). Similarly, Takafumi Yukawa et al. modified these quinolone compounds to generate a class of 1,8-naphthyridin-2-one compounds, such as compound 37 (Yukawa et al., 2020). Xiaodong Deng et al. screened and designed a class of novel 2-benzyloxyphenylacetonitrile derivatives, such as D2, through virtual screening and enzyme activity assays, which exhibited preliminary SMS2 inhibitory activity (Deng et al., 2014). Ya-li Li et al. identified a class of SAPA compounds, such as SAPA1a, through virtual screening, that exhibited good inhibitory activity against SMS1 (Li et al., 2015).Xiang-Yu Qi et al. synthesized a class of oxazolopyridine derivatives, such as QY16, with excellent SMS2 inhibitory activity and selectivity through scaffold hopping combined with molecular docking (Qi et al., 2017). Mingguang Mo et al. synthesized a class of benzisoxazole derivatives through scaffold hopping and ring closure and found that 15w, one of these compounds, exhibited excellent SMS2 inhibitory activity and selectivity, along with favorable pharmacokinetic properties. At an oral dose of 50 mg / kg, it significantly suppressed inflammatory cytokine levels in db / db mice, lowered LDL-C, and increased HDL-C levels (Mo et al., 2018). Yali Li et al. designed and obtained a class of salicylamide derivatives using electron isosteric methods and found that Ly93 in this class of compounds has high SMS2 inhibitory activity and selectivity, inhibiting liver SMS activity in a dose-dependent manner, reducing plasma SM levels, and ultimately inhibiting the development of atherosclerosis. In vitro results showed that Ly93 reduced apoB secretion in Huh7 cells, reduced SMS activity in macrophages, and increased cholesterol efflux in a dose-dependent manner. At the same time, Ly93 inhibited LPS-mediated secretion of proinflammatory cytokines and chemokines in macrophages (Li et al., 2019). Hadya Virupaksha Deepak et al. found that Daurichromenic acid (DCA), a compound isolated from the plant Rhododendron dauricum, inhibited SMS activity and showed inhibition of Aβ aggregation (EC). 50 =57 μM) (Deepak et al., 2020). Overall, although many SMS inhibitors have been discovered, the structure and drugability of SMS2 inhibitors still need to be further optimized.

[0006] Summary of the Invention

[0007] The first object of the present invention is to provide an alkoxythiophene carboxylic acid aromatic amide compound (including carboxyl / phosphate compounds derived therefrom as prodrugs) that has significant inhibitory activity against SMS2 and ideal physicochemical properties such as stability and water solubility.

[0008] The second object of the present invention is to provide a pharmaceutical composition containing an alkoxythiophene carboxylic acid aromatic amide compound (including a carboxylic acid / phosphate ester compound derived therefrom as a prodrug), or a pharmaceutically acceptable salt thereof.

[0009] The third object of the present invention is to provide pharmaceutical uses of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxyl / phosphate compounds derived therefrom as prodrugs) or pharmaceutically acceptable salts thereof.

[0010] In order to achieve the above-mentioned object, the present invention provides an alkoxythiophene carboxylic acid aromatic amine compound having a structure represented by formula (1), or a pharmaceutically acceptable salt thereof:

[0011] wherein X, Y, and Z are each independently selected from a carbon atom or a sulfur atom;

[0012] R 1 substituted or unsubstituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, thiazolyl, wherein the substituents of the substituted groups are 1 to 5 substituents selected from halogen, methyl, hydroxy, methoxy, trifluoromethyl, trifluoromethoxy, cyano, and nitro;

[0013] R 2 Selected from hydrogen, alkyl with less than 3 carbon atoms, and cycloalkyl;

[0014] R 3 is selected from substituted or unsubstituted hydrocarbon groups;

[0015] R 4 One or two selected from hydrogen, alkyl with less than 3 carbon atoms, halogen, and nitro.

[0016] As a preferred embodiment, X and Y are each independently selected from a sulfur atom or a carbon atom, and Z is a carbon atom.

[0017] As a preferred solution, the R 3 Select one of the following structures:

[0018] Wherein n is an integer from 1 to 10;

[0019] W is selected from a carbon atom or a nitrogen atom;

[0020] L and M are each independently selected from -C=C-, a sulfur atom, and an oxygen atom;

[0021] R 5 1 to 5 selected from hydrogen, substituted or unsubstituted alkyl groups with less than 4 carbon atoms, substituted or unsubstituted alkoxy groups with less than 20 carbon atoms, halogen, acyl, nitro, cyano, hydroxy, carboxylic (phosphonic) ester groups, and aromatic heterocycles.

[0022] As a preferred embodiment, the alkoxy group has less than 15 carbon atoms.

[0023] As a preferred embodiment, the compound is:

[0024] As a preferred embodiment, the pharmaceutically acceptable salt is hydrochloride, hydrobromide, tartrate, methanesulfonate, sodium salt, potassium salt, calcium salt, ammonium salt or amino acid salt.

[0025] The compounds of the present invention contain basic groups that can form salts with acids. Salts of derivatives can be formed using conventional means, including organic acid salts such as acetates, citrates, fumarates, maleates, oxalates, malates, citrates, succinates, tartrates, lactates, camphorsulfonates, benzenesulfonates, p-toluenesulfonates, methanesulfonates, trifluoroacetates, trifluoromethanesulfonates, etc.; inorganic acid salts such as hydrohalic acid salts (hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid), sulfates, phosphates, nitrates, etc.; or glutamate or aspartate salts can be formed with amino acids such as glutamic acid or aspartic acid, with preferred salts being hydrochloride or hydrobromide. Similarly, some compounds of the present invention contain acidic groups that can form salts with bases. Salts of derivatives can be formed using conventional means, including sodium salts, potassium salts, calcium salts, ammonium salts, or salts with amino acids such as lysine, arginine, histidine, etc., with preferred salts being sodium salts and potassium salts.

[0026] In order to achieve the second object of the present invention, the present invention provides a pharmaceutical composition comprising at least one therapeutically effective amount of an alkoxythiophene carboxylic acid aromatic amide compound represented by formula (1), or a pharmaceutically acceptable salt thereof.

[0027] The pharmaceutical composition of the present invention may also contain one or more pharmaceutically acceptable carriers, including conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field. If necessary, flavoring agents, sweeteners, etc. may also be added.

[0028] The pharmaceutical composition of the present invention can be prepared in any form, such as granules, powders, tablets, coated tablets, capsules, pills, syrups, drops, solutions, suspensions and emulsions, or sustained-release preparations of the active ingredient, wherein capsules include hard or soft gelatin capsules, and granules and powders can be non-effervescent or effervescent forms.

[0029] The pharmaceutical composition of the present invention can be administered by various routes according to conventional methods, including oral, intravenous, intraarterial, intraperitoneal, intrathoracic, transdermal, nasal, inhalation, rectal, ocular and subcutaneous administration.

[0030] In order to achieve the third object of the present invention, the present invention provides the use of the alkoxythiophene carboxylic acid aromatic amide compound or a pharmaceutically acceptable salt thereof in the preparation of a sphingomyelin synthase small molecule inhibitor.

[0031] The present invention uses a high-performance liquid chromatography (HPLC) fluorescence quantitative detection method reported in the literature to determine the inhibitory activity of the alkoxythiophene carboxylic acid aromatic amide compounds represented by formula (1) against sphingomyelin synthase (Deng et al., 2012). The change in the content of NBD-ceramide and NBD-sphingomyelin therein is used to calculate the change in the inhibitor's activity on sphingomyelin synthase-catalyzed conversion of ceramide to sphingomyelin. Activity testing experiments based on high-performance liquid chromatography (HPLC) fluorescence quantitative method show that the alkoxythiophene carboxylic acid aromatic amide compounds represented by formula (1) (including carboxyl / phosphate compounds derived therefrom as prodrugs) have micro- / submicromolar SMS2 inhibitory activity. The inhibitory activity of the compounds against SMS2 is detected by high-performance liquid chromatography (HPLC) fluorescence quantitative method.

[0032] The present invention also provides the use of the alkoxythiophene carboxylic acid aromatic amide compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating diseases caused by abnormally increased sphingomyelin levels.

[0033] As a preferred embodiment, the diseases caused by abnormally increased sphingomyelin levels include atherosclerosis, obesity, insulin resistance, fatty liver, type II diabetes, enteritis, pulmonary edema, lung damage, malignant tumors, Alzheimer's disease, and dry eye. The present invention also provides evaluation data on the therapeutic effect of pharmaceutical compositions containing the alkoxythiophene carboxylic acid aromatic amide compounds or pharmaceutically acceptable salts thereof on dry eye in mice.

[0034] The present invention has confirmed through experiments that the disclosed compounds have significant inhibitory activity against SMS2; ideal physical and chemical properties such as stability and water solubility; and they do not contain potentially toxic groups, with small potential toxic side effects. They can be used as drugs for preventing or treating diseases related to abnormal sphingomyelin levels, including metabolic syndrome related to metabolic disorders, such as atherosclerosis, obesity, insulin resistance, fatty liver, and type 2 diabetes; diseases related to inflammation, such as enteritis, pulmonary edema, and lung damage; malignant tumors, such as triple-negative breast cancer, colon cancer, leukemia, ovarian cancer, etc.; and central nervous system diseases, such as neurodegenerative diseases such as Alzheimer's disease, dry eye syndrome, etc.

[0035] The term "halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine and chlorine being preferred.

[0036] The term "alkyl" includes saturated aliphatic groups, including straight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.), branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl groups (cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl), alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In certain embodiments, the straight-chain or branched alkyl group has 4 or fewer carbon atoms in its backbone.

[0037] The term "alkoxy" includes substituted and unsubstituted alkyl groups covalently linked to an oxygen atom. In certain embodiments, a linear or branched alkoxy group has 20 or fewer carbon atoms in its backbone, more preferably 15 or fewer carbon atoms.

[0038] The beneficial effects of the present invention are that the alkoxythiophene carboxylic acid aromatic amide compounds provided by the present invention (including carboxyl / phosphate compounds derived therefrom as prodrugs) are a class of novel sphingomyelin synthase inhibitors with significant inhibitory activity against SMS2, ideal physical and chemical properties such as stability and water solubility, and do not contain potentially toxic groups, with low potential toxic side effects. They can be further prepared into drugs for preventing or treating diseases related to abnormal sphingomyelin levels.

[0039] Figures in the specification

[0040] Figure 1. Sodium fluorescein staining scores in mouse eyes. Pathological groups (including vehicle, cyclosporine, 1-2, and 1-31 groups) showed significantly increased staining scores compared to normal mice (physiological control group). In the former, both SMS2 inhibitors and cyclosporine reduced dry eye pathological scores. *p<0.05, **p<0.01, ****p<0.0001.

[0041] Figure 2. Sodium fluorescein staining of mouse eyes. Compared with normal mice (physiological control group), the pathological groups (including vehicle control group, cyclosporine, 1-2 and 1-31 groups) showed an increase in green area after staining, indicating that the degree of corneal damage also increased. Compared with the vehicle group, the SMS2 inhibitor cyclosporine and sporine both reduced the area of ​​corneal damage, and the effect of the former was more significant.

[0042] Figure 3 shows the tear secretion of mice. The cotton thread soaking length of the pathological groups (including vehicle, cyclosporine, 1-2, and 1-31 groups) was reduced compared with the normal mice (physiological control group), indicating a decrease in tear secretion. Compared with the vehicle group, the SMS2 inhibitors cyclosporine and cyclosporine both promoted tear secretion in mice, and the former was more significant (*p<0.05, ****p<0.0001). DETAILED DESCRIPTION

[0043] The technology of the present invention is described in detail below in conjunction with specific embodiments. It should be understood that the following specific embodiments are only intended to help those skilled in the art understand the present invention, and are not intended to limit the present invention.

[0044] Example 1: Preparation of 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-1)

[0045] 1) Synthesis of methyl 3-benzyloxythiophene-2-carboxylate

[0046] 5 g (0.03 mol, 1.0 eq) of compound 2, 13 g (0.09 mol, 3.0 eq) of potassium carbonate, and 500 mg of potassium iodide were weighed into a 250 mL round-bottom flask and dissolved in 100 mL of acetonitrile. 8.1 g (0.045 mol, 1.5 eq) of compound 3 was added with stirring and stirred at room temperature overnight. The reaction was complete after TLC detection. The mixture was filtered and evaporated to dryness to obtain 7.4 g of a yellow oil with a yield of 94%. MS (ESI) (m / z): 249 [M+H] + It can be used in the next reaction without further purification.

[0047] 2) Synthesis of 3-benzyloxythiophene-2-carboxylic acid

[0048] 7 g (0.03 mol, 1.0 eq) of compound 4 was weighed into a 250 mL round-bottom flask. 100 mL of a hydroxide solution (2 mol / L, H2O / dioxane = 1 / 1) was added at room temperature, and the mixture was refluxed in an oil bath for 8 h. The reaction was complete by TLC. The mixture was allowed to cool, and the pH was adjusted to neutral with dilute hydrochloric acid. The mixture was extracted with EA three times. The EA phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and recrystallized with PE / EA = 10 / 1 to obtain 6 g of yellow crystals with a yield of 85%. MS (ESI) (m / z): 233 [MH] - .

[0049] 3) Synthesis of 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Compound 1-1) (General Synthesis Method 1)

[0050] 3 g (0.013 mol, 1.0 eq) of compound 5 and 7.3 g (0.019 mol, 1.5 eq) of HBTU were weighed into a 100 mL round-bottom flask and dissolved in 50 mL of DMF. 4.95 g (0.038 mol, 3.0 eq) of DIPEA was added with stirring, and the mixture was stirred at room temperature for 10 min. 3.6 g (0.038 mol, 3.0 eq) of compound 6 was added and stirred at room temperature overnight. The reaction was complete by TLC. The mixture was dissolved in EA, washed three times with saturated brine, dried over anhydrous sodium sulfate, and flash purified to obtain 3.6 g of a yellow solid, 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-1), in an 89% yield. MS (ESI) (m / z): 311 [M+H]. + . 1 H NMR (400MHz, DMSO-d6) δ9.47 (s, 1H), 8.58 (s, 1H), 8.28 (m, 1H), 8.04 (d, J = 8.2Hz, 1H), 7.8 6(d,J=5.5Hz,1H),7.59(d,J=7.1Hz,2H),7.41(m,4H),7.28(d,J=5.4Hz,1H),5.44(s,2H).

[0051] Example 2: Preparation of Compounds 1-4, 1-5, 1-6, 1-7, 1-9, 1-14, 1-15, 1-20, 1-40, 1-61, 1-64, 1-65, 1-68, 1-69, and 1-83

[0052] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-aminopyridazine was reacted with Compound 5 to produce 3-benzyloxy-N-(pyridazin-3-yl)thiophene-2-carboxamide (Formula 1-4) in 50% yield. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.29(d,1H),8.61(d,1H),7.76(d,1H),7.60(t,1H),7.47–7.35(m,4H),7.27(s,1H),6.59(m,1H),5.20(s,2H).

[0053] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-4-trifluoromethylpyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(4-trifluoromethylpyridin-2-yl)thiophene-2-carboxamide (Formula 1-5) in a 16% yield. MS (ESI) (m / z): 379 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.97 (s, 1H), 8.64 (s, 1H), 8.43 (s, 1H), 7.53–7.33 (m, 5H), 7.27 (s, 1H), 7.23 (s, 1H), 6.91 (d, J = 4.5Hz, 1H), 5.40 (s, 2H).

[0054] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-4-chloropyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(4-chloropyridin-2-yl)thiophene-2-carboxamide (Formula 1-6) in a 46% yield. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.84(s,1H),8.43(s,1H),8.18(s,1H),7.56–7.33(m,5H),7.27(s,1H),7.03(s,1H),6.89(s,1H),5.39(s,2H).

[0055] Referring to the conditions and steps of Synthesis Method 1 in Example 1, aniline was reacted with Compound 5 to produce 3-benzyloxy-N-phenylthiophene-2-carboxamide (Formula 1-7) in a 68% yield. MS (ESI) (m / z): 310 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20(s,1H),7.49(m,5H),7.43(m,2H),7.28(s,2H),7.27–7.25(m,1H),7.07(s,1H),6.99(s,1H),5.29(s,2H).

[0056] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-6-hydroxypyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(6-hydroxypyridin-2-yl)thiophene-2-carboxamide (Formula 1-9) in a 48% yield. MS (ESI) (m / z): 327 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.53 (s, 1H), 7.76 (d, J = 7.5Hz, 1H), 7.58 (t, J = 7.5Hz, 1H), 7.43 (dd, J = 7.5, 1.5Hz, 2H) ,7.37(m,2H),7.27(m,1H),7.03(dd,J=7.5,1.4Hz,1H),6.68–6.51(m,2H),6.51–6.45(m,1H),5.20(s,2H).

[0057] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxy-5-nitrothiophene-2-carboxylic acid was reacted with Compound 6 to produce 3-benzyloxy-5-nitro-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-14) in a 56% yield. MS (ESI) (m / z): 356 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.16(s,1H),8.34(s,1H),8.20(m,2H),7.78(s,1H),7.50(m,2H),7.25(m,4H),5.32(s,2H).

[0058] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxy-4-bromothiophene-2-carboxylic acid was reacted with Compound 6 to produce 3-benzyloxy-4-bromo-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-15) in a 32% yield. MS (ESI) (m / z): 388, 390 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.03(s,1H),8.29(s,1H),7.99(s,2H),7.55–7.39(m,5H),7.24(s,1H),7.19(s,1H),5.35(s,2H).

[0059] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-4,6-dichloropyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(4,6-dichloropyridin-2-yl)thiophene-2-carboxamide (Formula 1-20) in a 7% yield. MS (ESI) (m / z): 379,381 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.90 (s, 1H), 8.34 (s, 1H), 7.50 (d, J = 15.8Hz, 5H), 7.41 (s, 1H), 7.28 (s, 1H), 7.06 (s, 1H), 6.94 (s, 1H), 5.38 (s, 2H).

[0060] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxy-5-methylthiophene-2-carboxylic acid was reacted with Compound 6 to produce 3-benzyloxy-5-methyl-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-40) in a 55% yield. MS (ESI) (m / z): 325 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.14 (s, 1H), 8.22 (d, J = 29.9Hz, 3H), 7.48 (m, 5H), 7.20 (m, 1H), 6.71 (s, 1H), 5.23 (s, 2H), 2.51 (s, 3H).

[0061] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-aminopyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(pyridin-2-yl)thiophene-2-carboxamide (Formula 1-61) in a 68% yield. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.78 (s, 1H), 8.31 (d, J = 7.8Hz, 2H), 7.70 (s, 1H), 7.43 (m, 5H), 7.28 (s, 1H), 7.02 (s, 1H), 6.88 (s, 1H), 5.40 (s, 2H).

[0062] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 5-aminopyrimidine was reacted with Compound 5 to prepare 3-benzyloxy-N-(pyrimidin-5-yl)thiophene-2-carboxamide (Formula 1-64) in a 43% yield. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.58(s,1H),8.98(m,2H),8.91(s,1H),7.89(s,1H),7.59(m,2H),7.42(m,3H),7.26(s,1H),5.45(s,2H).

[0063] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 5-aminothiazole was reacted with Compound 5 to produce 3-benzyloxy-N-(thiazol-5-yl)thiophene-2-carboxamide (Formula 1-65) in a 48% yield. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ 10.42 (s, 1H), 8.62 (s, 1H), 7.86 (s, 1H), 7.71 (s, 1H), 7.55 (m, 2H), 7.40 (d, J = 24.8Hz, 3H), 7.24 (s, 1H), 5.45 (s, 2H).

[0064] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-methyl-3-aminopyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(2-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-68) in a 53% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.37(s,1H),8.15(s,1H),7.89(s,1H),7 .59(m,2H),7.43(m,3H),7.34(s,1H),7.21(s,1H),5.42(s,2H),1.96(s,3H).

[0065] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-amino-4-methylpyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(4-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-69) in a 53% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.38 (s, 1H), 8.84 (s, 1H), 8.21 (s, 1H), 7.55–7.42 (m, 6H), 7.00 (d, J = 11.6Hz, 2H), 5.26 (s, 2H), 1.73 (s, 3H).

[0066] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-amino-5-methylpyridine was reacted with Compound 5 to produce 3-benzyloxy-N-(5-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-83) in a 72% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.41(s,1H),8.42(s,1H),8.13(s,1H),7.86(s,1H),7 .78(s,1H),7.60(m,2H),7.44(s,3H),7.28(s,1H),5.43(s,2H),2.28(s,3H).

[0067] Example 3: Preparation of 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2)

[0068] 1) Synthesis of 3-hydroxy-N-(pyridin-3-yl)thiophene-2-carboxamide

[0069] 2.3 g of compound 1-1 was weighed into a 100 mL round-bottom flask and dissolved in 30 mL of anhydrous dichloromethane. 1 mL of boron tribromide was added with stirring and stirred at room temperature overnight to precipitate a large amount of yellow solid. TLC showed that the reaction was complete. An appropriate amount of methanol was added to quench the dissolution, evaporated to dryness, and an appropriate amount of water was added. The pH was adjusted to alkaline with saturated sodium carbonate solution, acidified with dilute hydrochloric acid, extracted with EA, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 1 g of a yellow solid with a yield of 65%. MS (ESI) (m / z): 221 [M+H] + This compound was used in the next reaction without further purification.

[0070] 2) Synthesis of 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2) (General Synthesis Method 2)

[0071] 70 mg (0.32 mmol, 1.0 eq) of compound 7 and 88 mg (0.64 mmol, 2.0 eq) of potassium carbonate were weighed into a round-bottom flask and dissolved in 3 mL of NMP. 92 mg (0.413 mmol, 1.3 eq) of compound 8 was added with stirring and stirred at room temperature overnight. The reaction was complete as determined by TLC. EA was added for dissolution, the mixture was washed three times with saturated brine, and dried over anhydrous sodium sulfate. Flash purification was performed to obtain 75 mg of a white solid, 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2), in a 65% yield. MS (ESI) (m / z): 363, 365 [M+H]. + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.62(s,1H),8.29(m,1H),8.05(s,1H),7.89(s,1H),7.65(m,2H),7.34(m,2H),5.48(s,2H).

[0072] Example 4: Compounds 1-3, 1-8, 1-10, 1-11, 1-12, 1-13, 1-16, 1-18, 1-19, 1-21, 1-22, 1-23, 1-25, 1-27, 1-28, 1-29, 1-30, 1-31, 1-32, 1-33, 1-34, 1-35, 1-36, 1-37, 1-38, 1-39, 1- Preparation of 41, 1-42, 1-43, 1-44, 1-45, 1-46, 1-47, 1-48, 1-49, 1-50, 1-51, 1-52, 1-53, 1-54, 1-55, 1-56, 1-57, 1-58, 1-59, 1-60, 1-63, 1-66, 1-67, 1-70, 1-71, 1-72, 1-73, 1-74, 1-75, 1-76, 1-77, 1-78, 1-79, 1-80, 1-81, 1-82, 1-84, 1-85, 1-86, 1-87, 1-88, 1-89, 1-93, 1-94, 1-97, 1-98, 1-99

[0073] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,6-dichlorobenzyl bromide was reacted with Compound 7 to produce 3-(2,6-dichlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-3) in a 75% yield. MS (ESI) (m / z): 379, 381 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.22(s,1H),8.36(d,J=2.3Hz,1H),8.27(d,J=4.6Hz,1H),7.95(d,J=5.6Hz,2H) ,7.64(s,1H),7.62(s,1H),7.53(m,1H),7.45(d,J=5.5Hz,1H),7.36(dd,J=8.2,4.7Hz,1H),5.60(s,2H).

[0074] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 5-fluoro-2-(2-hydroxy-2-methylpropoxy)benzyl bromide was reacted with Compound 7 to produce 3-(5-fluoro-2-(2-hydroxy-2-methylpropoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-8) in a 44% yield. MS (ESI) (m / z): 416 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.98(s,1H),8.51(d,J=1.3Hz,1H),8.24(dd,J=7.5,1.3Hz,1H),8.06(d,J=7.5Hz,1H),7.77(d,J=7.3Hz,1H ),7.30–7.22(m,2H),7.10–7.01(m,2H),6.97(m,1H),6.59(d,J=7.5Hz,1H),5.21(s,2H),4.25(s,2H),2.19(s,1H),1.35(s,6H).

[0075] Referring to the conditions and steps of Synthesis Method 2 in Example 3, naphthalen-1-ylmethyl bromide was reacted with Compound 7 to produce 3-(naphthalen-1-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-10) in a 68% yield. MS (ESI) (m / z): 361 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.28(s,1H),8.35(d,J=7.5Hz,1H),8.21(s,1H),8.15(s,1H),8.01(s,2H ),7.91(s,1H),7.80(d,J=6.1Hz,1H),7.73(s,1H),7.61–7.44(m,4H),7.27(s,1H),5.93(s,2H).

[0076] Referring to the conditions and steps of Synthesis Method 2 in Example 3, naphthalene-2-ylmethyl bromide was reacted with Compound 7 to produce 3-(naphthalene-2-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-11) in a 69% yield. MS (ESI) (m / z): 361 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.53(s,1H),8.61(s,1H),8.25(s,1H),8.14–7.79(m,6H),7.71(s,1H),7.52(s,2H),7.30(d,J=11.6Hz,2H),5.59(s,2H).

[0077] Referring to the conditions and steps of Synthesis Method 2 in Example 3, quinolin-8-ylmethyl bromide was reacted with Compound 7 to produce N-(pyridin-3-yl)-3-(quinolin-8-ylmethoxy)thiophene-2-carboxamide (Formula 1-12) in a 78% yield. MS (ESI) (m / z): 362 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.57(s,1H),8.88(s,1H),8.46(d,J=8.1Hz,1H),8.27(d,J=14.2Hz,2H),8.05(d,J=7.6Hz ,2H),7.86(s,2H),7.68(d,J=7.1Hz,1H),7.57(d,J=3.7Hz,1H),7.43(s,1H),7.29(d,J=24.7Hz,1H),6.03(s,2H).

[0078] Referring to the conditions and steps of Synthesis Method 2 in Example 3, anthracene-9-ylmethyl bromide was reacted with Compound 7 to produce 3-(anthracen-9-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-13) in a 55% yield. MS (ESI) (m / z): 411 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.03(s,1H),8.82(s,1H),8.59(d,J=8.0Hz,2H),8.21(d,J=8.0Hz,2H),8. 10(s,1H),8.00(s,1H),7.74(s,1H),7.59(s,5H),7.36(d,J=7.2Hz,1H),7.13(s,1H),6.50(s,2H).

[0079] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-methoxynaphthalen-1-ylmethyl bromide was reacted with Compound 7 to produce 3-((2-methoxynaphthalen-1-yl)methoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-16) in an 88% yield. MS (ESI) (m / z): 391 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.23(s,1H),8.27(d,J=8.2Hz,1H),8.19(d,J=3.9Hz,1H),8.09(d,J=9.0Hz,1H),8.02(s,1H),7.93(dd,J=16.0,6.8Hz ,2H),7.66(d,J=7.1Hz,1H),7.56(d,J=9.0Hz,1H),7.54–7.46(m,2H),7.39(d,J=7.4Hz,1H),7.26(d,J=4.0Hz,1H),5.91(s,2H),3.97(s,3H).

[0080] Referring to the conditions and steps of Synthesis Method 2 in Example 3, ethyl 6-(2-(bromomethyl)phenoxy)hexanoate was reacted with Compound 7 to produce ethyl 6-(2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)phenoxy)hexanoate (Formula 1-18) in a 32% yield. MS (ESI) (m / z): 469 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.43(d,J=14.0Hz,1H),8.34(d,J=8.2Hz,1H),8.25(d,J= 11.2Hz,1H),7.95(d,J=7.5Hz,1H),7.88(d,J=10.8Hz,1H),7.55(s,1H),7.46–7.3 0(m,3H),7.11(d,J=7.5Hz,1H),7.01(s,1H),5.37(s,2H),3.95(dd,J=32.5,10.5H z, 4H), 2.14 (s, 2H), 1.57 (s, 2H), 1.40 (s, 2H), 1.25 (s, 2H), 1.14 (t, J = 2.7Hz, 3H).

[0081] Referring to the conditions and steps of Synthesis Method 2 in Example 3, (6,7,8,9-tetrafluorodibenzo[b,e][1,4]dioxol-1-yl)methyl bromide was reacted with Compound 7 to prepare N-(pyridin-3-yl)-3-((6,7,8,9-tetrafluorodibenzo[b,e][1,4]dioxol-1-yl)methoxy)thiophene-2-carboxamide (Formula 1-19) in a 45% yield. MS (ESI) (m / z): 489 [M+H] + . 1 HNMR(400MHz,DMSO-d6)δ9.41(s,1H),8.52(s,1H),8.21(s,1H),7.99(s,1 H),7.90(s,1H),7.39–7.26(m,3H),7.22(s,1H),7.17(s,1H),5.43(s,2H).

[0082] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-nitrobenzyl bromide was reacted with Compound 7 to produce 3-(2-nitrobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-21) in a 16% yield. MS (ESI) (m / z): 356 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.43(s,1H),8.69(s,1H),8.44–8.02(m,3H),7.84(m,3H),7.66(s,1H),7.38(s,1H),7.24(s,1H),5.80(s,2H).

[0083] Referring to the conditions and steps of Synthesis Method 2 in Example 3, methyl 2-(bromomethyl)benzoate was reacted with Compound 7 to produce methyl 2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)benzoate (Formula 1-22) in a 45% yield. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.61(s,1H),8.27(s,1H),8.05(d,J=7.7Hz,1H),7.97(d,J=7.5Hz,1H),7.85(d,J=5.0Hz,1H) ,7.75(d,J=7.4Hz,1H),7.67(t,J=7.3Hz,1H),7.54(t,J=7.2Hz,1H),7.37(s,1H),7.25(d,J=5.3Hz,1H),5.72(s,2H),3.74(s,3H).

[0084] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-chloro-5-(trimethylacetoxy)benzyl bromide was reacted with Compound 7 to prepare trimethylacetic acid (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol ester (Formula 1-23) in a 36% yield. MS (ESI) (m / z): 445, 447 [M+H]. + . 1 H NMR(400MHz, DMSO-d6)δ9.43(s,1H),8.61(d,J=3.1Hz,1H),8.26(s,1H),8.00(s,1H),7.88(t,J=5.6Hz,1H ),7.58(dd,J=12.6,3.9Hz,2H),7.33(d,J=5.5Hz,2H),7.24–7.15(m,1H),5.45(s,2H),1.26–1.20(s,9H).

[0085] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-chloro-5-(diethoxyphosphoryloxy)benzyl bromide was reacted with Compound 7 to produce diethyl 4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy)phenyl phosphate (Formula 1-25) in 85% yield. MS (ESI) (m / z): 497,499 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45(s,1H),8.64(s,1H),8.29(s,1H),8.04(s,1H),7.89(s,1H),7.59(d,J=10.6Hz,2 H),7.37(s,1H),7.30(t,J=7.2Hz,2H),5.50(d,J=8.7Hz,2H),4.08(dd,J=15.2,7.5Hz,4H),1.22–1.14(m,6H).

[0086] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(dodecyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(dodecyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-27) in a 68% yield. MS (ESI) (m / z): 495 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.39 (s, 1H), 8.26 (m, 2H), 8.15 (s, 1H), 7.46 (d, J = 34.6Hz, 3H), 7.24–7 .18(m,1H),7.03(s,3H),5.31(s,2H),3.95(m,2H),1.65(m,2H),1.23(m,18H),0.88(t,3H).

[0087] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(Undecyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(Undecyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-28) in a 52% yield. MS (ESI) (m / z): 481 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.39 (s, 1H), 8.28 (m, 2H), 8.15 (s, 1H), 7.47 (d, J = 33. 6Hz,3H),7.26–7.17(m,1H),7.04(m,3H),5.42(s,2H),3.96(m,2H),1.66(m, 2H),1.22(m,16H),0.88(t,3H).

[0088] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(decyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(decyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-29) in a 48% yield. MS (ESI) (m / z): 467 [M+H] + . 1 H NMR(400MHz, CDCl3) δ9.43(d,J=18.0Hz,1H),8.31(d,J=18.7Hz,2H),8.19(d,J=18.3Hz,1H),7.60–7.40(m,3H), 7.28(d,J=5.6Hz,2H),7.08(d,J=18.7Hz,3H),5.32(s,2H),3.98(m,2H),1.68(m,2H),1.24(m,14H),0.92(t,3H).

[0089] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(nonyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(nonyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-30) in a 58% yield. MS (ESI) (m / z): 453 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(m,2H),8.15(s,1H),7.50(s,1H),7.42(m,2H),7.25(s,1H),7.2 3(s,1H),7.08–6.96(m,3H),5.31(s,2H),3.95(d,J=4.9Hz,2H),1.65(m,2H),1.22(m,12H),0.86(t,3H).

[0090] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(octyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(octyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-31) in a 59% yield. MS (ESI) (m / z): 439 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(s,2H),8.15(s,1H),7.50(s,1H),7.42(s,2H),7.25–7.16(m,1H),7. 08–6.95(m,3H),5.31(s,2H),3.96(s,2H),1.65(d,J=5.9Hz,2H),1.35–1.13(m,10H),0.86(t,J=6.7Hz,3H).

[0091] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(heptyloxy)benzyl bromide was reacted with Compound 7 to prepare 3-((2-(heptyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-32) in a 52% yield. MS (ESI) (m / z): 425 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.38(s,1H),8.26(d,J=8.2Hz,2H),8.15(s,1H),7.51(d,J=5.0Hz,1H),7.42(m,2H),7.23(s,1H) ,7.02(dd,J=22.1,7.0Hz,3H),5.29(s,2H),3.96(t,J=6.1Hz,2H),1.70–1.61(m,2H),1.38–1.11(m,8H),0.83(t,3H).

[0092] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(hexyloxy)benzyl bromide was reacted with Compound 7 to produce 3-((2-hexyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-33) in a 56% yield. MS (ESI) (m / z): 411 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.39(s,1H),8.27(m,2H),8.15(s,1H),7.51(s,1H),7.42(m,2H),7. 24(m,2H),7.05(m,3H),5.32(s,2H),3.96(m,2H),1.65(m,2H),1.25(m,6H),0.81(t,3H).

[0093] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(pentyloxy)benzyl bromide was reacted with Compound 7 to produce 3-((2-pentyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-34) in a 21% yield. MS (ESI) (m / z): 397 [M+H] + .1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(m,2H),8.15(s,1H),7.51(s,1H),7.42(m,2H),7.24–7.19 (m,1H),7.02(d,J=18.8Hz,3H),5.32(s,2H),3.96(m,2H),1.65(m,2H),1.26(m,4H),0.81(t,3H).

[0094] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(butoxy)benzyl bromide was reacted with Compound 7 to produce 3-((2-butoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-35) in a 30% yield. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(s,2H),8.16(s,1H),7.52(s,1H),7.43(2,2H),7.25–7.18 (m,1H),7.02(d,J=20.3Hz,3H),5.37(s,2H),3.97(m,2H),1.65(m,2H),1.35(m,2H),0.84(t,3H).

[0095] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(propyloxy)benzyl bromide was reacted with Compound 7 to produce 3-((2-propyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-36) in a 91% yield. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.39(s,1H),8.27(s,2H), 8.17(s,1H),7.52(s,1H),7.42(m,2H),7.25–7.16(m,1H),7.03(d,J=22.0Hz,3H),5.33(s,2H),3.93(m,2H),1.69(m,2H),0.91(t,3H).

[0096] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(ethoxy)benzyl bromide was reacted with Compound 7 to produce 3-((2-ethoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-37) in a 57% yield. MS (ESI) (m / z): 355 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.40 (s, 1H), 8.27 (s, 2H), 8.19 (s, 1H), 7.52 (s, 1H), 7.42 (d, J = 6. 7Hz,2H),7.25–7.18(m,1H),7.11–6.90(m,3H),5.38(s,2H),4.04(m,2H),1.31(t,3H).

[0097] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-n-pentyloxy-6-chlorobenzyl bromide was reacted with Compound 7 to produce 3-(2-n-pentyloxy-6-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-38) in a 57% yield. MS (ESI) (m / z): 430, 432 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.32(s,1H),8.27(d,J=2.8Hz,2H),8.17(d,J=1.8Hz,1H),7.51(d,J=5.4Hz,1H),7.34(d,J=8.2Hz,1H),7.24(d ,J=7.8Hz,1H),7.13(dd,J=16.8,6.7Hz,2H),6.90(d,J=8.1Hz,1H),5.50(s,2H),3.97(m,2H),1.71(m,2H),1.30(m,4H),0.84(t,3H).

[0098] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-chloro-6-n-propoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-chloro-6-n-propoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-39) in a 52% yield. MS (ESI) (m / z): 403, 405 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.32(s,1H),8.28(m,2H),8.18(s,1H),7.51(s,1H),7.34(s,1H),7.25–7.20 (m,1H),7.14(d,J=15.8Hz,2H),6.90(s,1H),5.51(s,2H),3.94(m,2H),1.75(m,2H),0.94(t,3H).

[0099] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-methylbenzyl bromide was reacted with Compound 7 to produce 3-(2-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-41) in a 91% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.48(s,1H),8.28(s,1H),8.00(d,J=7.0Hz,1H),7. 89(s,1H),7.54(d,J=6.3Hz,1H),7.30(dd,J=21.5,7.0Hz,5H),5.46(s,2H),2.40(s,3H).

[0100] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-methylbenzyl bromide was reacted with Compound 7 to produce 3-(4-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-42) in a 60% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.46(s,1H),8.58(s,1H),8.29(s,1H),8.04(d,J=7.1Hz,1H),7.86(d,J=5.3Hz,1 H), 7.48 (d, J = 7.4Hz, 2H), 7.38 (d, J = 5.3Hz, 1H), 7.26 (dd, J = 15.0, 6.2Hz, 3H), 5.39 (s, 2H), 2.31 (s, 3H).

[0101] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-methylbenzyl bromide was reacted with Compound 7 to produce 3-(3-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-43) in 84% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.49 (s, 1H), 8.58 (s, 1H), 8.28 (d, J = 4.7Hz, 1H), 8.05 (d, J = 8.3Hz, 1H), 7.87 (d, J = 5. 5Hz, 1H), 7.43 (s, 1H), 7.33 (ddd, J = 27.2, 12.9, 6.4Hz, 4H), 7.20 (d, J = 7.0Hz, 1H), 5.39 (s, 2H), 2.31 (s, 3H).

[0102] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-fluorobenzyl bromide was reacted with Compound 7 to produce 3-(2-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-44) in a 67% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.42(s,1H),8.57(s,1H),8.29(d,J=4.4Hz,1H),8.03(d,J=8.0Hz,1H), 7.89(d,J=5.5Hz,1H),7.70(t,J=7.4Hz,1H),7.58–7.43(m,1H),7.44–7.23(m,4H),5.52(s,2H).

[0103] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-fluorobenzyl bromide was reacted with Compound 7 to produce 3-(3-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-45) in a 54% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.34(s,1H),8.50(s,1H),8.15(s, 1H), 7.94 (s, 1H), 7.72 (s, 1H), 7.31 (d, J = 22.9Hz, 4H), 7.10 (m, 2H), 5.31 (d, J = 7.5Hz, 2H).

[0104] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-fluorobenzyl bromide was reacted with Compound 7 to produce 3-(4-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-46) in a 41% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.60 (s, 1H), 8.29 (s, 1H), 8.04 (d, J = 7.4Hz, 1H), 7.8 6(d,J=5.2Hz,1H),7.67(m,2H),7.38(d,J=4.7Hz,1H),7.27(t,J=7.5Hz,3H),5.42(s,2H).

[0105] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-chlorobenzyl bromide was reacted with Compound 7 to produce 3-(2-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-47) in a 48% yield. MS (ESI) (m / z): 345, 347 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ9.42(s,1H),8.55(s,1H),8.29(d,J=4.1Hz,1H),8.03(d,J=8.1Hz,1H),7. 90(d,J=5.5Hz,1H),7.74(d,J=6.8Hz,1H),7.58(d,J=7.3Hz,1H),7.51–7.30(m,4H),5.52(s,2H).

[0106] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-chlorobenzyl bromide was reacted with Compound 7 to produce 3-(3-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-48) in a 56% yield. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.49(s,1H),8.67(s,1H),8.29(s,1H),8.07(d,J=8.2Hz,1H),7.87(d,J=5.5Hz, 1H),7.73(s,1H),7.54(s,1H),7.51–7.42(m,2H),7.41–7.32(m,1H),7.25(d,J=5.3Hz,1H),5.44(s,2H).

[0107] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-chlorobenzyl bromide was reacted with Compound 7 to produce 3-(4-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-49) in a 63% yield. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.46 (s, 1H), 8.63 (s, 1H), 8.29 (d, J = 4.2Hz, 1H), 8.06 (d, J = 8.3Hz, 1H), 7.86 (d, J = 5. 5Hz,1H),7.62(d,J=8.1Hz,2H),7.50(d,J=8.1Hz,2H),7.44–7.35(m,1H),7.25(d,J=5.5Hz,1H),5.43(s,2H).

[0108] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-cyanobenzyl bromide was reacted with Compound 7 to produce 3-(2-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-50) in a 51% yield. MS (ESI) (m / z): 336 [M+H] +. 1 H NMR (400MHz, CDCl3) δ9.01 (s, 1H), 8.32 (d, J = 13.9Hz, 3H), 7.65 (m, 5H), 7.25 (m, 1H), 7.00 (s, 1H), 5.50 (s, 2H).

[0109] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-cyanobenzyl bromide was reacted with Compound 7 to produce 3-(3-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-51) in a 54% yield. MS (ESI) (m / z): 336 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.01(s,1H),8.31(m,2H),8.23(s,1H),7.77(m,3H),7.63(s,1H),7.53(s,1H),6.93(s,1H),5.34(s,2H).

[0110] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-cyanobenzyl bromide was reacted with Compound 7 to produce 3-(4-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-52) in a 46% yield. MS (ESI) (m / z): 336 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.02(s,1H),8.34(d,J=14.2Hz,2H),8.23(s,1H),7.78(d,J=8.0Hz,2H), 7.61 (d, J=7.3Hz, 2H), 7.52 (d, J=5.4Hz, 1H), 7.27 (s, 1H), 6.90 (d, J=5.5Hz, 1H), 5.38 (s, 2H).

[0111] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-trifluoromethoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-53) in a 51% yield. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.09 (s, 1H), 8.27 (m, 1H), 7.60 (d, J = 7.3Hz, 3H), 7.52 (s, 1H), 7.41 (m, 2H), 7.29 (m, 2H), 7.00 (s, 1H), 5.39 (s, 2H).

[0112] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-trifluoromethoxybenzyl bromide was reacted with Compound 7 to produce 3-(4-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-54) in a 70% yield. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.12(s,1H),8.41(s,1H),8.32(s,1H),8.13(s,1H),7.53(m,3H),7.26(m,3H),6.96(s,1H),5.30(s,2H).

[0113] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-trifluoromethoxybenzyl bromide was reacted with Compound 7 to produce 3-(3-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-55) in a 55% yield. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.10 (s, 1H), 8.32 (m, 2H), 8.18 (s, 1H), 7.49 (d, J = 29.7Hz, 3H), 7.34 (m, 2H), 7.23 (s, 1H), 6.96 (s, 1H), 5.31 (s, 2H).

[0114] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-methoxybenzyl bromide was reacted with Compound 7 to produce 3-(4-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-56) in a 23% yield. MS (ESI) (m / z): 341 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.24 (s, 1H), 8.26 (d, J = 17.2Hz, 3H), 7.51 (s, 1H), 7.41 (s, 2H), 7.20 (m, 1H), 6.99 (s, 3H), 5.21 (s, 2H), 3.85 (s, 3H).

[0115] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-methoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-57) in a 78% yield. MS (ESI) (m / z): 341 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.39(s,1H),8.28(s,1H),8.22(m,2H),7.51(s,1H),7.4 2(s,1H),7.23(m,2H),7.02(dd,J=20.4,6.7Hz,3H),5.32(s,2H),3.82(s,3H).

[0116] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-methoxybenzyl bromide was reacted with Compound 7 to produce 3-(3-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-58) in a 46% yield. MS (ESI) (m / z): 341 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.23(s,1H),8.29(m,3H),7.45(m,2H),7.29(s,1H),7.00(m,4H),5.26(s,2H),3.83(s,3H).

[0117] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-trifluoromethylbenzyl bromide was reacted with Compound 7 to produce 3-(2-trifluoromethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-59) in a 48% yield. MS (ESI) (m / z): 379 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.09 (s, 1H), 8.26 (m, 3H), 7.66 (dd, J = 81.3, 36.6Hz, 5H), 7.18 (m, 1H), 6.97 (s, 1H), 5.51 (s, 2H).

[0118] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,6-dimethylbenzyl bromide was reacted with Compound 7 to produce 3-(2,6-dimethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-60) in a 55% yield. MS (ESI) (m / z): 339 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.19(s,1H),8.24(s,1H),8.11(s,1H),7.91(s,1H),7. 56(s,1H),7.28(s,1H),7.18(m,3H),7.10(s,1H),5.34(s,2H),2.46(s,6H).

[0119] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-chloro-6-methoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-chloro-6-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-63) in a 45% yield. MS (ESI) (m / z): 375, 377 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.37(s,1H),8.32(m,2H),8.26(s,1H),7.55(s,1H),7.41(s,1H) ,7.27(m,1H),7.18(d,J=8.0Hz,2H),6.96(d,J=7.9Hz,1H),5.55(s,2H),3.90(s,3H).

[0120] Referring to the conditions and steps of Synthesis Method 2 in Example 3, benzo[b]thiophen-3-ylmethyl bromide was reacted with Compound 7 to produce 3-(Benzo[b]thiophen-3-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-66) in a 34% yield. MS (ESI) (m / z): 367 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.99(s,1H),8.20(s,1H),7.98(s,1H),7.88(d,J=8.8Hz,2H ),7.76(s,1H),7.63(s,1H),7.56(s,1H),7.40(m,2H),7.09(m,2H),5.53(s,2H).

[0121] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,3,5,6-tetrafluorobenzyl bromide was reacted with Compound 7 to produce 3-(2,3,5,6-tetrafluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-67) in a 65% yield. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.98 (s, 1H), 8.49 (s, 1H), 8.35 (d, J = 3.6Hz, 1H), 8.26 (d, J = 8 .2Hz,1H),7.54(d,J=5.5Hz,1H),7.29(d,J=8.0Hz,1H),7.23–7.14(m,1H),7.07(d, J=5.5Hz,1H),5.49(s,2H).

[0122] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(phenethoxy)benzyl bromide was reacted with Compound 7 to produce 3-(2-(phenethoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-70) in 80% yield. MS (ESI) (m / z): 431 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.30(s,1H),8.27(m,2H),8.11(s,1H),7.52(s,1H),7.40(m,2H),7 .25–7.19(m,1H),7.05(dd,J=45.4,17.6Hz,8H),5.25(s,2H),4.20(t,2H),2.96(t,2H).

[0123] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-chloro-2-hexyloxybenzyl bromide was reacted with Compound 7 to produce 3-(4-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-71) in a 60% yield. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.27(s,1H),8.28(d,J=12.0Hz,3H),7.51(s,1H),7.45–7.32(m,2H),7.26–7.17(m, 1H), 7.00 (s, 1H), 6.92 (d, J = 8.6Hz, 1H), 5.26 (s, 2H), 3.94 (m, 2H), 1.66 (m, 2H), 1.25 (m, 6H), 0.82 (t, 3H).

[0124] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 4-chloro-2-heptyloxybenzyl bromide was reacted with Compound 7 to produce 3-(4-chloro-2-heptyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-72) in a 70% yield. MS (ESI) (m / z): 459, 461 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.25(s,1H),8.26(d,J=7.6Hz,3H),7.51(s,1H),7.38(d,J=15.2Hz,2H),7.25(s,2 H), 7.00 (s, 1H), 6.90 (d, J = 7.7Hz, 1H), 5.26 (s, 2H), 3.93 (m, 2H), 1.65 (m, 2H), 1.23 (m, 8H), 0.83 (t, 3H).

[0125] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 5-chloro-2-hexyloxybenzyl bromide was reacted with Compound 7 to produce 3-(5-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-73) in a 15% yield. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.27 (s, 1H), 8.28 (d, J = 10.5Hz 3H),7.52(s,1H),7.40(d,J=14.8Hz,3H),7.01(s,1H),6.93(s,1H),5.26(s,2H),3.95(m,2H),1.66(m,2H),1.26(m,6H),0.82(t,3H).

[0126] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 5-chloro-2-heptyloxybenzyl bromide was reacted with Compound 7 to produce 3-(5-chloro-2-heptyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-74) in a 56% yield. MS (ESI) (m / z): 459, 461 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.07(s,1H),8.07(m,3H),7.32(s,1H),7.22(m,2H),7.08(s,1H),6. 81(s,1H),6.73(s,1H),5.07(s,2H),3.74(m,2H),1.47(m,2H),1.05(m,8H),0.65(t,3H).

[0127] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(4-chlorobutoxy)benzyl bromide was reacted with Compound 7 to produce 3-(2-(4-chlorobutoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-75) in a 48% yield. MS (ESI) (m / z): 417, 419 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.43(s,1H),8.34(s,1H),8.19(s,1H),7.60(s,1H),7.50–7.41(m,1H),7 .38–7.19(m,2H),7.09(d,J=30.2Hz,2H),5.49(s,2H),4.08(m,2H),3.58(m,2H),1.90(m,4H).

[0128] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(6-chlorohexyloxy)benzyl bromide was reacted with Compound 7 to produce 3-(2-(6-chlorohexyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-76) in a 47% yield. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.40 (d, J = 47.7Hz, 1H), 8.27 (d, J = 57.3Hz, 3H), 7.60 (s, 1H), 7.32 (d, J = 16.6 Hz,2H),7.09(d,J=23.1Hz,3H),5.46(s,2H),4.04(m,2H),3.46(m,2H),1.73(m,4H),1.42(m,4H).

[0129] Referring to the conditions and steps of Synthesis Method 2 in Example 3, thiophen-2-ylmethyl bromide was reacted with Compound 7 to produce 3-(thiophen-2-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-77) in a 9% yield. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.18 (s, 1H), 8.32 (d, J = 19.4Hz, 3H), 7.51 (d, J = 27.6Hz, 2H), 7.26–7.16 (m, 2H), 7.06 (d, J = 26.8Hz, 2H), 5.45 (s, 2H).

[0130] Referring to the conditions and steps of Synthesis Method 2 in Example 3, thiophen-3-ylmethyl bromide was reacted with Compound 7 to produce 3-(thiophen-3-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-78) in 8% yield. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.21(s,1H),8.30(m,3H),7.60–7.42(m,3H),7.30(m,2H),6.99(s,1H),5.30(s,2H).

[0131] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-hexyloxybenzyl bromide was reacted with Compound 7 to produce 3-(3-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-79) in a 12% yield. MS (ESI) (m / z): 411 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.24(s,1H),8.31(m,2H),8.20(s,1H),7.51(s,1H),7.38(s,1H),7.27(s,1H),7.23(s,1H), 7.06(s,1H),6.98(d,J=8.1Hz,3H),5.24(s,2H),3.95(m,2H),1.76(m,2H),1.43(m,2H),1.32(m,4H),0.89(t,3H).

[0132] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-chloro-2-hexyloxybenzyl bromide was reacted with Compound 7 to produce 3-(3-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-80) in a 54% yield. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20(s,1H),8.41(s,1H),8.27(d,J=28.6Hz,2H),7.50(d,J=14.1Hz,2H),7.37(s,1H ),7.16(m,2H),7.02(s,1H),5.31(s,2H),4.04(m,2H),1.75(m,2H),1.38(m,2H),1.22(m,4H),0.82(t,3H).

[0133] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-ethylbenzyl bromide was reacted with Compound 7 to produce 3-(2-ethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-81) in 82% yield. MS (ESI) (m / z): 339 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20 (s, 1H), 8.26 (d, J = 4.3Hz, 1H), 8.17 (d, J = 7.8Hz, 1H), 8.07 (s, 1H), 7.54 (d, J = 4.5Hz, 1H), 7.44 (t, J = 7.8Hz, 2H),7.34(dd,J=16.6,7.1Hz,2H),7.24–7.18(m,1H),7.04(d,J=4.4Hz,1H),5.31(s,2H),2.79(q,J=7.2Hz,2H),1.26(t,J=7.2Hz,3H).

[0134] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-isopropoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-isopropoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-82) in a 43% yield. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.37(s,1H),8.37–8.17(m,3H),7.52(d,J=5.5Hz,1H),7.43(dd,J=15.9,6.9Hz,2H),7.2 3(dd,J=8.2,4.9Hz,1H),7.07–6.97(m,2H),5.32(s,2H),4.61(dt,J=12.0,5.9Hz,1H),1.27(d,J=6.0Hz,6H).

[0135] Referring to the conditions and steps of Synthesis Method 2 in Example 3, ethyl 2-bromomethylbenzoate was reacted with Compound 7 to produce ethyl 2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)benzoate (Formula 1-84) in a 53% yield. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.94 (s, 1H), 8.24 (dd, J = 7.5, 1.3Hz, 1H), 8.08 (dt, J = 7.5 ,1.4Hz,1H),8.02(d,J=1.3Hz,1H),7.89(d,J=7.5Hz,1H),7.77(dd,J=7.4,1.4H z,1H),7.64(td,J=7.5,1.4Hz,1H),7.56–7.46(m,2H),7.26(t,J=7.5Hz,1H),6 .55(d,J=7.5Hz,1H),5.20(s,2H),4.31(q,J=6.0Hz,2H),1.38(t,J=6.0Hz,3H).

[0136] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-difluoromethoxybenzyl bromide was reacted with Compound 7 to produce 3-(2-difluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-85) in a 47% yield. MS (ESI) (m / z): 377 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.16 (s, 1H), 8.27 (m, 3H), 7.53 (d, J = 15.4Hz, 3H), 7.34 (s,1H),7.25–7.16(m,2H),7.01(s,1H),6.60(t,J=73.2Hz,1H),5.37(s,2H).

[0137] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-bromobenzyl bromide was reacted with Compound 7 to produce 3-(2-bromobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-86) in a 36% yield. MS (ESI) (m / z): 389, 391 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.17 (s, 1H), 8.30 (m, 3H), 7.71 (d, J = 7.8Hz, 1H), 7.52 (m, 2H), 7.42 (s, 1H), 7.33 (s, 2H), 7.01 (s, 1H), 5.38 (s, 2H).

[0138] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-iodobenzyl bromide was reacted with Compound 7 to produce 3-(oxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-87) in a 39% yield. MS (ESI) (m / z): 436, 438 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.14(s,1H),8.28(m,3H),7.98(s,1H),7.57–7.41(m,3H),7.25–7.21(m,1H),7.17(s,1H),7.00(s,1H),5.32(s,2H).

[0139] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 5-fluoro-2-methylbenzyl bromide was reacted with Compound 7 to produce 3-(5-fluoro-2-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-88) in a 38% yield. MS (ESI) (m / z): 343 [M+H] + . 1 HNMR(400MHz, CDCl3)δ9.11(s,1H),8.30(s,1H),8.21(m,2H),7.54(s,1H),7.28(s,1H),7. 26–7.21(m,1H),7.16(d,J=8.4Hz,1H),7.07(s,1H),6.99(s,1H),5.26(s,2H),2.40(s,3H).

[0140] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,5-dichlorobenzyl bromide was reacted with Compound 7 to produce 3-(2,5-dichlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-89) in a 48% yield. MS (ESI) (m / z): 379, 381 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.07 (s, 1H), 8.37 (d, J = 33.1Hz, 3H), 7.54 (m, 2H), 7.42 (d, J = 27.6Hz, 2H), 6.98 (s, 1H), 5.37 (s, 2H).

[0141] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,5-dipropoxybenzyl bromide was reacted with Compound 7 to produce 3-(2,5-dipropoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-93) in a 90% yield. MS (ESI) (m / z): 427 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.36 (s, 1H), 8.25 (d, J = 14.6Hz, 3H), 7.53 (d, J = 29.3Hz, 1H), 7.24 (s, 1H), 7 .11–6.86(m,4H),5.29(s,2H),3.87(m,4H),1.77(m,2H),1.68(m,2H),1.01(t,3H),0.90(t,3H).

[0142] Referring to the conditions and steps of Synthesis Method 2 in Example 3, (1,1'-biphenyl-2-yl)methyl bromide was reacted with Compound 7 to produce 3-((1,1'-biphenyl-2-yl)methoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-94) in a 41% yield. MS (ESI) (m / z): 387 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.45(s,1H),8.28(s,1H),7.96(s,1H),7.79(m,2H),7.49(m,2H),7.39(m,7H),6.97(s,1H),5.36(s,2H).

[0143] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 3-trifluoromethylbenzyl bromide was reacted with Compound 7 to produce 3-(3-trifluoromethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-97) in a 43% yield. MS (ESI) (m / z): 379 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ9.50 (s, 1H), 8.65 (s, 1H), 8.28 (s, 1H), 8.03 (m, 2H), 7.88 (d, J = 4.2Hz, 2H), 7.75 (d, J=7.1Hz, 1H), 7.69 (d, J=7.3Hz, 1H), 7.37 (dd, J=7.9, 4.4Hz, 1H), 7.29 (d, J=4.9Hz, 1H), 5.52 (s, 2H).

[0144] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2,5-dimethoxybenzyl bromide was reacted with Compound 7 to produce 3-(2,5-dimethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-98) in a 52% yield. MS (ESI) (m / z): 371 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.51(s,1H),8.54(s,1H),8.29(s,1H),8.01(d,J=8.5Hz,1H),7.87(d,J=5.5Hz,1H ),7.46–7.32(m,2H),7.17(s,1H),7.03(d,J=9.1Hz,1H),6.96(d,J=8.9Hz,1H),5.38(s,2H),3.71(s,6H).

[0145] Referring to the conditions and steps of Synthesis Method 2 in Example 3, adamantan-1-ylmethyl bromide was reacted with Compound 7 to produce 3-(adamantan-1-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-99) in a 98% yield. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.29(s,1H),8.72(s,1H),8.31(s,1H),8.15(s,1H),7.86(d,J=5.3 Hz,1H),7.52–7.38(m,1H),7.21(d,J=5.4Hz,1H),3.91(s,2H),2.00(m,3H),1.67(m,12H).

[0146] Example 5: Preparation of 3-benzyloxy-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90)

[0147] 1) Synthesis of 3-benzyloxythiophene-2-carbonyl chloride

[0148] 1 g (4.27 mmol, 1.0 eq) of compound 5 was weighed into a round-bottom flask and dissolved in 50 mL of DCM. 5.4 g (42.7 mmol, 10.0 eq) of oxalyl chloride was added with stirring in an ice bath, followed by 500 μL of DMF as catalyst. The mixture was stirred at room temperature for 7 h. TLC confirmed the reaction was complete, and the mixture was evaporated to dryness. EA was added, the mixture was washed once with saturated brine, and dried over anhydrous sodium sulfate. Evaporation gave 996 mg of a pale yellow solid, a yield of 92%. This product was used in the next reaction without further purification.

[0149] 2) Synthesis of 3-benzyloxy-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90) (General Synthesis Method 3)

[0150] 71 mg (0.64 mmol, 2.0 eq) of compound 10 was weighed into a round-bottom flask and dissolved in 4 mL of DCM. 82 mg (0.64 mmol, 2.0 eq) of DIPEA was added, and 80 mg (0.32 mmol, 1.0 eq) of compound 9 was added with stirring. The mixture was stirred at room temperature overnight. The reaction was complete as determined by TLC, and the reaction was stopped. Flash purification was performed to obtain 100 mg of a white solid, 3-(benzyloxy)-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90), in a 96% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.45(s,1H),8.89(s,1H),7.82(s,1H),7.53–7.39(m,5H),7.26–7.23(m,1H),7.16(s,1H),6.98(s,1H),5.31(s,2H).

[0151] Example 6: Preparation of Compounds 1-62, 1-91, and 1-92

[0152] Referring to the conditions and steps of Synthesis Method 3 in Example 5, 3-methylaminopyridine was reacted with Compound 9 to produce 3-benzyloxy-N-methyl-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-62) in a 27% yield. MS (ESI) (m / z): 325 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.35(d,J=7.5Hz,2H),7.34(t,J=11.0Hz,4H),7.23(d,J=5.4Hz,1 H), 7.19 (d, J = 6.6Hz, 2H), 7.07 (s, 1H), 6.50 (d, J = 4.2Hz, 1H), 4.73 (s, 2H), 3.44 (s, 3H).

[0153] Referring to the conditions and steps of Synthesis Method 3 in Example 5, 3-amino-4-chloropyridine was reacted with Compound 9 to prepare 3-benzyloxy-N-(4-chloropyridin-3-yl)thiophene-2-carboxamide (Formula 1-91) in a 26% yield. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.79(s,1H),9.51(s,1H),8.20(s,1H),7.54–7.40(m,5H),7.26(s,1H),7.20(s,1H),6.99(s,1H),5.28(s,2H).

[0154] Referring to the conditions and steps of Synthesis Method 3 in Example 5, 3-amino-5-fluoropyridine was reacted with Compound 9 to prepare 3-benzyloxy-N-(5-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-92) in a 98% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.36 (s, 1H), 8.22 (d, J = 9.7Hz, 1H), 8.16 (s, 1H), 7.89 (s ,1H),7.56(d,J=5.2Hz,1H),7.51(m,5H),7.02(d,J=4.8Hz,1H),5.30(s,2H).

[0155] Example 7: Preparation of Compounds 1-95 and 1-96

[0156] 50 mg (0.23 mmol, 1.0 eq) of compound 7 and 63 mg (0.45 mmol, 2.0 eq) of potassium carbonate were weighed into a round-bottom flask and dissolved in 3 mL of NMP. 50 mg of compound 11 was added with stirring, and the mixture was transferred to a 70°C oil bath and stirred overnight. TLC confirmed the reaction was complete, and the mixture was dissolved in EA. The mixture was washed three times with saturated brine, dried over anhydrous sodium sulfate, and flash purified to yield 30 mg of a yellow solid in a 48% yield. MS (ESI) (m / z): 277 [M+H]. + . 1H NMR(400MHz,DMSO-d6)δ9.40(s,1H),8.75(s,1H),8.31(s,1H),8.12(s,1H),7.87(s ,1H),7.42(s,1H),7.22(s,1H),4.33(m,2H),1.85(m,2H),1.49(m,2H),0.97(t,3H).

[0157] Referring to the preparation conditions and steps of compound 1-95, 8-iodooctane was reacted with compound 7 to prepare 3-octyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-96) in a 54% yield. MS (ESI) (m / z): 333 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.39(s,1H),8.74(s,1H),8.30(s,1H),8.10(s,1H),7.86(s,1H ),7.39(s,1H),7.21(s,1H),4.30(m,2H),1.84(m,2H),1.49–1.18(m,10H),0.80(t,3H).

[0158] Example 8: Preparation of Compounds 1-17 and 1-24

[0159] 100 mg of compound 1-18 was weighed into a 100 mL round-bottom flask. 20 mL of a 2 mol / L hydroxide solution (H₂O / dioxane = 1 / 1) was added at room temperature. The mixture was refluxed in an oil bath for 8 h. The reaction was complete by TLC. The mixture was cooled and the pH was adjusted to neutral with dilute hydrochloric acid. The mixture was extracted three times with EA. The EA phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, and flash purified to yield 80 mg of a white solid, 6-(2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)phenoxy)hexanoic acid (Formula 1-17), in 85% yield. MS (ESI) (m / z): 441 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.36 (s, 1H), 8.26 (d, J = 4.4Hz, 1H), 7.95 (d, J = 7 .5Hz,1H),7.87(d,J=5.4Hz,1H),7.53(d,J=7.3Hz,1H),7.43–7.28(m,3H),7.08(d,J =8.2Hz,1H),7.00(t,J=7.3Hz,1H),5.37(s,2H),3.92(t,J=6.0Hz,2H),2.07(t,J=7. 3Hz, 2H), 1.55 (d, J = 6.4Hz, 2H), 1.37 (dd, J = 14.4, 7.2Hz, 2H), 1.25 (d, J = 6.1Hz, 2H).

[0160] Referring to the preparation conditions and steps of compound 1-17, 3-(2-chloro-5-hydroxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-24) was prepared using compound 1-23 as a reactant in a 69% yield. MS (ESI) (m / z): 361, 363 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.88(s,1H),9.43(s,1H),8.59(s,1H),8.29(d,J=4.4Hz,1H),8.05(d,J=8.2Hz,1H),7.89(d,J=5.4Hz, 1H), 7.38 (dd, J=8.0, 4.8Hz, 1H), 7.33 (d, J=8.6Hz, 1H), 7.28 (d, J=5.4Hz, 1H), 7.08 (s, 1H), 6.81 (d, J=8.6Hz, 1H), 5.43 (s, 2H).

[0161] Example 9: (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol phosphate (Formula 1-26)

[0162] 373 mg (0.75 mmol, 1.0 eq) of compound 1-25 was weighed into a round-bottom flask and dissolved in 12 mL of acetonitrile. 1.15 g (7.5 mmol, 10.0 eq) of TMS-Br was added with stirring. The mixture was stirred at room temperature for 48 h, resulting in the precipitation of a large amount of white solid. Acetonitrile was added and ultrasonic filtration was performed to obtain 246 mg of a white solid, (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol phosphate (Formula 1-26), in a 74% yield. MS (ESI) (m / z): 441, 443 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.87(s,1H),9.14(s,1H),8.57(s,1H),8.40(d,J=8.2Hz,1H),7.95(d,J=5.3Hz,1H),7.8 7(d,J=4.6Hz,1H),7.53(d,J=8.6Hz,1H),7.49(s,1H),7.32(d,J=5.2Hz,1H),7.24(d,J=8.5Hz,1H),5.50(s,2H).

[0163] Example 10: Preparation of the hydrochloride salt of compound formula 1-2

[0164] 0.60 g (1.66 mmol, 1.0 eq) of compound 1-2 was dissolved in 10 mL of dry ethyl acetate. 1.44 mL (1.8 mmol, 1.1 eq) of HCl (g) in ethyl acetate (c = 1.25 mol / L) was added dropwise to the above solution under ice-water bath conditions. After reacting for 10 min, the solution was filtered and dried to obtain 0.53 g of a white powdery solid with a yield of 81%.

[0165] Example 11: Preparation of Compound Formula 1-100

[0166] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-(benzyloxy)thiophene-3-carboxylic acid and 3-aminopyridine were used as starting materials to obtain 4-(benzyloxy)-N-(pyridin-3-yl)thiophene-3-carboxamide (Formula 1-100) via amide condensation in an 80% yield. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR(400MHz,dmso)δ9.89(s,1H),8.63(s,1H),8.27(s,1H),8.17(s,1H),8.07(d, J=8.0Hz,1H),7.57(d,J=7.2Hz,2H),7.46–7.33(m,4H),6.94(s,1H),5.21(s,2H)

[0167] Comparative Example: Preparation of Compounds 12-1 to 12-20

[0168] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-(2-ethylbenzyloxy)thiophene-2-carboxylic acid was reacted with Compound 6 to produce 2-(2-ethylbenzyloxy)-N-(pyridin-3-yl)benzamide (Formula 12-1) in a 72% yield. MS (ESI) (m / z): 333 [M+H] + . 1HNMR(400MHz,DMSO-d6)δ11.06(s,1H),9.18(d,J=1.6Hz,1H),8.60(d,J=5.3Hz,1H) ,8.44(d,J=8.5Hz,1H),7.94(dd,J=8.5,5.4Hz,1H),7.64(dd,J=7.6,1.6Hz,1H),7.6 1–7.55(m,1H),7.49(d,J=7.4Hz,1H),7.40(d,J=8.3Hz,1H),7.26(dt,J=13.9,6.5H z, 2H), 7.19–7.10 (m, 2H), 5.27 (s, 2H), 2.68 (q, J = 7.5Hz, 2H), 1.12 (t, J = 7.5Hz, 3H).

[0169] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-aminopyridazine was reacted with Compound 5 to prepare 3-benzyloxy-N-(pyridazin-4-yl)thiophene-2-carboxamide (Formula 12-2) in a 96% yield. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.74(s,1H),9.19(d,J=1.8Hz,1H),9.05(d,J=5.9Hz,1H),7.97(dd,J=5.9,2.7Hz,1H),7 .94(d,J=5.5Hz,1H),7.60(s,1H),7.59(s,1H),7.41(dt,J=22.3,7.2Hz,3H),7.29(d,J=5.5Hz,1H),5.46(s,2H).

[0170] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-1,3,4-thiadiazole was reacted with Compound 5 to prepare 3-benzyloxy-N-(1,3,4-thiadiazol-2-yl)thiophene-2-carboxamide (Formula 12-3) in a 33% yield. MS (ESI) (m / z): 318 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.65(s,1H),8.84(s,1H),7.57(s,1H),7.45(m,4H),7.27(s,1H),6.92(s,1H),5.41(s,2H).

[0171] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-4-fluoropyridine was reacted with Compound 5 to prepare 3-benzyloxy-N-(4-fluoropyridin-2-yl)thiophene-2-carboxamide (Formula 12-4) in a 38% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.89 (s, 1H), 8.23 ​​(s, 1H), 8.13 (d, J = 9.1Hz, 1H), 7.45 (d, J = 21.3Hz, 3H), 7.27 (s, 1H), 6.89 (s, 1H), 6.77 (s, 1H), 5.39 (s, 2H).

[0172] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(1,3-dioxoisoindol-2-yl)benzyl bromide was reacted with Compound 7 to prepare 3-(2-(1,3-dioxoisoindol-2-yl)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 12-5) in a 50% yield. MS (ESI) (m / z): 456 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.97(s,1H),8.44(m,1H),8.34(m,1H),8.13(s,1H),7.87(m,1H),7.79–7.72 (m,4H),7.61(s,1H),7.49–7.37(m,2H),7.32(t,1H),7.24–7.19(m,1H),6.63(s,1H),5.24(s,2H).

[0173] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-aminodibenzo[b,d]furan was reacted with Compound 5 to prepare 3-(benzyloxy)-N-(dibenzo[b,d]furan-3-yl)thiophene-2-carboxamide (Formula 12-6) in 75% yield. MS (ESI) (m / z): 400 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.65 (s, 1H), 8.12 (s, 1H), 8.06 (t, J = 6.9Hz, 2H), 7.88 ( d,J=5.4Hz,1H),7.66(dd,J=17.7,7.5Hz,3H),7.51–7.35(m,5H),7.31(d,J=5.2 Hz, 1H), 7.24 (d, J = 8.2Hz, 1H), 5.46 (s, 2H).

[0174] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-9H-fluorene was reacted with Compound 5 to prepare 3-(benzyloxy)-N-(9H-fluoren-2-yl)thiophene-2-carboxamide (Formula 12-7) in a 38% yield. MS (ESI) (m / z): 398 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ9.48(s,1H),7.83(dd,J=11.6,6.7Hz,3H),7.77(s,1H),7.62(d,J=7.0 Hz, 2H), 7.56 (d, J = 7.0 Hz, 1H), 7.52–7.39 (m, 4H), 7.39–7.23 (m, 3H), 5.45 (s, 2H), 3.90 (s, 2H).

[0175] Referring to the conditions and steps of Synthesis Method 2 in Example 3, 2-(2-((5-methylpyridin-2-yl)amino)-2-oxoethyl)benzyl bromide was reacted with Compound 7 to prepare 3-(2-(2-((5-methylpyridin-2-yl)amino)-2-oxoethyl)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 12-8) in a 63% yield. MS (ESI) (m / z): 459 [M+H]. + . 1 H NMR (400MHz, DMSO-d6) δ10.66(s,1H),9.26(s,1H),8.42(s,1H),8.24(s,1H),8.05(s,1H),7.93(d,J=8.3Hz,1H),7.84(s,1H) ,7.74(d,J=8.7Hz,1H),7.58(s,1H),7.47–7.38(m,3H),7.31(dd,J=21.7,5.8Hz,3H),5.48(s,2H),3.95(s,2H),2.19(s,3H).

[0176] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-(benzyloxy)-2-methyl-N-(pyridin-3-yl)thiazole-5-carboxamide (Formula 12-9) was prepared by reacting 4-(benzyloxy)-2-methyl-N-(pyridin-3-yl)thiazole-5-carboxamide with Compound 6 in a 77% yield. MS (ESI) (m / z): 326 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ9.19(s,1H),8.46(s,1H),8.12(s,1H),7.85(d,J=8.0Hz,1 H), 7.41 (d, J = 6.9 Hz, 2H), 7.23 (dd, J = 14.6, 6.8 Hz, 4H), 5.41 (s, 2H), 2.50 (s, 3H).

[0177] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3,4,5-trifluoroaniline was reacted with Compound 5 to prepare 3-(benzyloxy)-N-(3,4,5-trifluorophenyl)thiophene-2-carboxamide (Formula 12-10) in 50% yield. MS (ESI) (m / z): 364 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.24 (s, 1H), 7.55 (m, 5H), 7.32 (s, 1H), 7.06 (d, J = 21.9Hz, 3H), 5.44 (s, 2H).

[0178] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-aminopyridine was reacted with Compound 5 to produce 3-(benzyloxy)-N-(pyridin-4-yl)thiophene-2-carboxamide (Formula 12-11) in a 97% yield. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.30(s,1H),8.41(m,2H),7.50(m,5H),7.26(m,3H),6.99(s,1H),5.28(s,2H).

[0179] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-amino-6-methylpyridine was reacted with Compound 5 to produce 3-(benzyloxy)-N-(6-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 12-12) in a 94% yield. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.38(s,1H),8.43(s,1H),7.91(d,J=8.1Hz,1H),7.85(s,1H),7 .58(m,2H),7.44(s,3H),7.28(s,1H),7.22(d,J=8.7Hz,1H),5.42(s,2H),2.41(s,3H).

[0180] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-amino-6-fluoropyridine was reacted with Compound 5 to produce 3-(benzyloxy)-N-(6-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 12-13) in a 20% yield. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.23 (s, 1H), 8.26 (s, 1H), 7.85 (s, 1H), 7.51 (d, J = 19.4Hz, 5H), 7.27 (s, 1H), 7.00 (s, 1H), 6.88 (s, 1H), 5.28 (s, 2H).

[0181] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-amino-6-trifluoromethylpyridine was reacted with Compound 5 to produce 3-(benzyloxy)-N-(6-trifluoromethylpyridin-3-yl)thiophene-2-carboxamide (Formula 12-14) in a 48% yield. MS (ESI) (m / z): 379 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.76 (s, 1H), 8.73 (s, 1H), 8.34 (d, J = 7.4Hz, 1H), 7.90 (m, 2H),7.59(d,J=6.4Hz,2H),7.47–7.37(m,3H),7.28(d,J=5.2Hz,1H),5.45(s,2H).

[0182] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxypyridine-2-carboxylic acid was reacted with Compound 6 to prepare 3-benzyloxy-N-(pyridin-3-yl)pyridine-2-carboxamide (Formula 12-15) in a 43% yield. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.67(s,1H),8.87(d,J=2.5Hz,1H),8.28(dd,J=4.7,1.5Hz,1H),8.26–8.21(m,1H),8.18(d,J=8.1Hz,1 H),7.75–7.67(m,1H),7.54(dd,J=8.5,4.6Hz,1H),7.46(d,J=7.2Hz,2H),7.41–7.31(m,3H),7.29(d,J=7.2Hz,1H),5.25(s,2H).

[0183] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-benzyloxypyridine-3-carboxylic acid was reacted with Compound 6 to prepare 4-benzyloxy-N-(pyridin-3-yl)pyridine-3-carboxamide (Formula 12-16) in a 30% yield. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.61(d,J=2.4Hz,1H),8.53(s,1H),8.45(d,J=5.8Hz,1H),8.16(d,J=4.8Hz,1H), 7.99(d,J=8.4Hz,1H),7.39(d,J=7.3Hz,1H),7.31(t,J=6.0Hz,1H),7.26–7.18(m,4H),7.07–7.01(m,1H),5.19(s,2H).

[0184] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxypyridine-4-carboxylic acid was reacted with Compound 6 to prepare 3-benzyloxy-N-(pyridin-3-yl)pyridine-4-carboxamide (Formula 12-17) in a 40% yield. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.69(d,J=34.6Hz,2H),8.32(dd,J=22.3,4.6Hz,2H),8 .09(d,J=8.2Hz,1H),7.52(dd,J=28.8,5.9Hz,3H),7.35(dd,J=16.5,9.5Hz,4H),5.34(s,2H).

[0185] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 2-benzyloxypyridine-3-carboxylic acid was reacted with Compound 6 to produce 2-benzyloxy-N-(pyridin-3-yl)pyridine-3-carboxamide (Formula 12-18) in a 36% yield. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ10.46(s,1H),8.74(d,J=2.6Hz,1H),8.42–8.34(m,1H),8.31(d,J=4.7Hz,1H),8 .12(d,J=7.3Hz,2H),7.54(d,J=7.2Hz,2H),7.43–7.28(m,4H),7.21(dd,J=7.3,4.9Hz,1H),5.49(s,2H).

[0186] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 4-benzyloxypyrimidine-5-carboxylic acid was reacted with Compound 6 to produce 4-benzyloxy-N-(pyridin-3-yl)pyrimidine-5-carboxamide (Formula 12-19) in a 60% yield. MS (ESI) (m / z): 307 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.56(s,1H),8.95(s,1H),8.85(s,1H),8.75(d,J=2.5Hz,1H),8.30( dd,J=4.7,1.5Hz,1H),8.13–8.06(m,1H),7.55–7.45(m,2H),7.41–7.30(m,4H),5.53(s,2H).

[0187] Referring to the conditions and steps of Synthesis Method 1 in Example 1, 3-benzyloxypiperazine-2-carboxylic acid was reacted with Compound 6 to prepare 3-benzyloxy-N-(pyridin-3-yl)piperazine-2-carboxamide (Formula 12-20) in a 60% yield. MS (ESI) (m / z): 307 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.73(s,1H),8.73(d,J=2.5Hz,1H),8.31(d,J=2.6Hz,1H),8.22(d,J=2.6Hz,1H),8.18(dd,J=4.7,1.4H z,1H),8.04(dd,J=8.5,1.9Hz,1H),7.34(d,J=7.2Hz,2H),7.30–7.25(m,1H),7.25–7.20(m,2H),7.19–7.14(m,1H),5.35(s,2H).

[0188] The structure of the comparative compound is as follows:

[0189] Example 12: Determination of the inhibitory effect of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxylic acid / phosphate compounds derived therefrom as prodrugs) on SMS2 in vitro

[0190] Experimental instruments and materials

[0191] 1. Electric constant temperature water bath (Shanghai Yiheng Technology Co., Ltd.).

[0192] 2. Vortex mixer (Shanghai Jingke Industrial Co., Ltd. Model XW-80A).

[0193] 3. High-speed centrifuge (model Eppendorf 5804R).

[0194] 4. High performance liquid chromatography Agilent 1100 (Agilent Technologies, Palo Alto, CA, USA) equipped with quaternary pump, vacuum degassing, and FLD fluorescence detector.

[0195] 5. HPLC column COSMOSIL 5C18-MS-II (4.6 mm ID x 250 mm).

[0196] 6. DMPC was purchased from Santa Cruz (USA) and dissolved in ethanol at a concentration of 40 mmol / L. C6-NBD-Ceramide was purchased from Santa Cruz (USA) and dissolved in DMSO at a concentration of 1.16 mmol / L.

[0197] 7. All organic solvents used were purchased from Shanghai Sinopharm Reagent Company. Methanol was chromatographic grade. Water was ultrapure water filtered by a Milli-Q pump, deionized, and ultrafiltered through a 0.22 μm membrane. Other biological consumables were purchased from domestic companies.

[0198] 8. Prepare the test compound solution: Accurately weigh 1-2 mg of each test compound and add an appropriate amount of DMSO to prepare a 10 mmol / L stock solution. Take a certain volume of the DMSO stock solution of the test compound and then add an appropriate amount of DMSO to dilute the test compound to the desired concentration.

[0199] 9.SMS2 pure enzyme, DDM solution and buffer were provided by Cao Yu's group at the National Center for Protein Science (Shanghai).

[0200] Detection of the inhibitory activity of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxyl / phosphate compounds derived therefrom as prodrugs) against SMS2

[0201] Take 6 μL of SMS2 protein (0.144 mg / mL) and dilute it in 3 mL of DDM buffer solution, then vortex. Take 79 μL of the above enzyme dilution solution and add it to a 1.5 mL EP tube. Add 1 μL of the inhibitor solution in DMSO and vortex for 15 seconds. Let it stand at room temperature for 5 minutes. Add 20 μL of a mixed solution of DMPC and C6-NBD-ceramide and vortex for 15 seconds. Incubate at 37°C for 30 minutes. Add 200 μL of anhydrous ethanol to quench the reaction and vortex for 15 seconds. Take out 200 μL and store it in the dark at 4°C for HPLC detection.

[0202] The fluorescence quantitative analysis of the samples prepared above was performed using the same high-performance liquid chromatography method as in the reference (Xiaodong Deng, et al. Analytical Letters, 2012, 45(12):1581-1589). The peak area A(sm) and A(ceramide) values ​​of C6-NBD-SM and C6-NBD-Ceramide in the DMSO and inhibitor groups were analyzed and recorded on the corresponding HPLC spectra. The inhibition rate of the test compound was calculated according to the following formula.

[0203] The inhibition rates of the target compounds (including comparative example compounds) on SMS2 at a concentration of 1 μM or 10 μM were measured as follows:

[0204] (Each set of data was measured twice and the average value was taken; “-” means not measured).

[0205] Example 13: Evaluation of the therapeutic effect of SMS2 inhibitors on dry eye in mice

[0206] (1) Experimental materials

[0207] Experimental animals: 6-8 week old male C57BL / 6 experimental mice.

[0208] Experimental instruments: Alisn constant temperature blast drying oven, desktop microscope, cobalt blue lamp.

[0209] Experimental reagents and consumables: phenol red cotton thread, 1% sodium fluorescein solution, Xingqi eye drops 0.05% cyclosporine eye drops (Ⅱ) 0.04 mg / g preparation of Example Compound 1-2, 0.04 mg / g preparation of Example Compound 1-31.

[0210] (2) Experimental methods

[0211] Twenty-eight male C57BL / 6 mice, aged 6-8 weeks, were housed in a constant-temperature forced-air drying oven at 19.5% humidity and 25°C for two weeks to establish a dry eye model. At the end of the two weeks, the dry eye mice were randomly divided into four groups of seven mice each. Subsequently, the humidity in the constant-temperature forced-air drying oven was adjusted to 50%, and the mice were housed for another week. Each group of mice received the corresponding drug treatment: the vehicle group received empty vehicle without the SMS2 inhibitor; the cyclosporine group received commercial cyclosporine eye drops; the Example Compound 1-2 group received 0.04% Compound 1-2; and the Example Compound 1-31 group received 0.04% Compound 1-31. Each mouse received 5 μl of pharmaceutical ocular topical solution twice daily. Seven other male C57BL / 6 mice, aged 6-8 weeks, were housed in a constant-temperature forced-air drying oven at 50% humidity and 25°C throughout the entire experiment. During the third week, they received empty vehicle as a physiological control.

[0212] At the end of the third week, ocular surface damage recovery was assessed using sodium fluorescein staining in all mice. Tear production was also measured using a phenol red thread. Under anesthesia, the phenol red thread was placed in the conjunctival sac of the mouse for 30 seconds, after which the length of the thread discoloration was measured. Subsequently, 2 μl of 1% sodium fluorescein solution was applied to the ocular surface, and images were taken under cobalt blue light for scoring.

[0213] Experimental results

[0214] The experimental results show that SMS2 inhibitors have significant therapeutic effects on the treatment of corneal epithelial damage caused by dry eye, and can also promote tear secretion in dry eye mice. Two SMS2 inhibitors, Example Compound 1-2 and Example Compound 1-31, both showed efficacy in the treatment of dry eye (Figures 1 and 2), as shown by a reduction in corneal sodium fluorescein staining scores, and showed statistically significant advantages compared with the commonly used commercial dry eye preparation cyclosporine eye drops in clinical practice. In terms of promoting tear secretion (Figure 3), cyclosporine and the two SMS2 inhibitors showed an upward trend compared with the vehicle group. These results indicate that SMS2 inhibitors have the potential to treat dry eye.

[0215] The above is only a preferred embodiment of the present invention. It should be pointed out that ordinary technicians in this technical field can make several improvements and modifications without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An alkoxythiophenecarboxylic acid aromatic amide compound of the structure represented by formula (1), or a pharmaceutically acceptable salt thereof: in, X, Y, and Z are each independently selected from a carbon atom or a sulfur atom; R 1 is selected from substituted or unsubstituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, thiazolyl, wherein the substituents of the substituted groups are selected from 1 to 5 of halogen, methyl, hydroxy, methoxy, trifluoromethyl, trifluoromethoxy, cyano, and nitro; R 2 Selected from hydrogen, alkyl with less than 3 carbon atoms, and cycloalkyl; R 3 is selected from substituted or unsubstituted hydrocarbon groups; R 4 One or two selected from hydrogen, alkyl with less than 3 carbon atoms, halogen, and nitro.

2. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The X and Y are each independently selected from a sulfur atom or a carbon atom, and the Z is a carbon atom.

3. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The R 3 Select one of the following structures: Wherein n is an integer from 1 to 10; W is selected from a carbon atom or a nitrogen atom; L and M are each independently selected from -C=C-, a sulfur atom, and an oxygen atom; R 5 1 to 5 selected from hydrogen, substituted or unsubstituted alkyl with less than 4 carbon atoms, substituted or unsubstituted alkoxy with less than 20 carbon atoms, halogen, acyl, nitro, cyano, hydroxyl, carboxylic (phosphonic) ester, and aromatic heterocycle.

4. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 3, or a pharmaceutically acceptable salt thereof, characterized in that: The alkoxy group is an alkoxy group with no more than 15 carbon atoms.

5. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound is:

6. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salt is hydrochloride, hydrobromide, tartrate, methanesulfonate, sodium salt, potassium salt, calcium salt, ammonium salt or amino acid salt.

7. A pharmaceutical composition, characterized in that Contains at least one therapeutically effective amount of an alkoxythiophenecarboxylic acid aromatic amide compound represented by formula (1) as described in any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof.

8. Use of the alkoxythiophene carboxylic acid aromatic amide compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, in the preparation of a small molecule inhibitor of sphingomyelin synthase.

9. Use of the alkoxythiophene carboxylic acid aromatic amide compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for preventing and treating a disease caused by an abnormal increase in sphingomyelin levels.

10. The use of claim 9 in the preparation of a drug for preventing and treating a disease caused by an abnormal increase in sphingomyelin levels, characterized in that: The diseases caused by abnormal increase in sphingomyelin levels include atherosclerosis, obesity, insulin resistance, fatty liver, type II diabetes, enteritis, pulmonary edema, lung damage, malignant tumors, Alzheimer's disease and dry eye.

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