A compound, its preparation method and use in the preparation of sEH inhibitors and PPARs agonists

Abstract

The application belongs to the technical field of medicine, and particularly relates to a compound, a preparation method thereof and application of the compound in preparation of sEH inhibitors and PPARs agonists. The application provides a compound with a structure shown in formula V or VI. The compound provided by the application has a typical urea structure as a primary pharmacophore of soluble epoxide hydrolase (sEH) and a thiazolidinedione part as a primary pharmacophore of peroxisome proliferator-activated receptors (PPARs). The sEH inhibitor and PPARs agonist compound provided by the application has high inhibitory activity on human HsEH and high agonistic activity on PPARs, and can be used as an sEH inhibitor and PPARs agonist compound for preparation of a medicine for treating diseases mediated by soluble epoxide hydrolase and peroxisome proliferator-activated receptors.

Description

[0001] This application is a divisional application of Chinese patent application filed on March 22, 2023, with application number 202310282598.7, entitled "A dual-target compound and its preparation method and its application in the preparation of sEH inhibitors and PPARs agonists". Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, specifically relating to a compound, its preparation method, and its application in the preparation of sEH inhibitors and PPARs agonists. Background Technology

[0003] Pain sensation is mediated by a specialized subset of sensory afferent neurons (nociceptors), which are activated in response to thermal, mechanical, and chemical stimuli through various mechanisms. Studies have shown that ion channel regulation, including transient receptor potential (TRP) channels, G protein-coupled receptor (GPCR) activation, and cell membrane alterations, demonstrates the role of lipid mediators in signal transduction within nociceptors (Nature, 2001, 413(6852):203-210).

[0004] Studies have shown that cyclooxygenase and lipoxygenase metabolites, prostaglandins and leukotrienes, can lead to pain and inflammation, demonstrating the role of lipid mediators in pain signal transduction. Specific long-chain polyunsaturated fatty acids (PUFAs) are metabolized by cytochrome P450 enzymes (CYP450) to form epoxide metabolites, namely epoxidized fatty acids (EpFAs). Researchers have found that these metabolites mediate analgesic effects in several types of pain pathology, such as acute pain, chronic pain, cancer pain, or intractable pain.

[0005] Arachidonic acid (ARA) is a 20-carbon PUFA containing four unsaturated double bonds. It can be metabolized by CYP450 enzymes into epoxide metabolites (EETs) of any one or more of these four double bonds, including 5,6-EET, 8,9-EET, 11,12-EET, and 14,15-EET. EpFAs, including EETs, limit pain and inflammation through a variety of direct and indirect mechanisms, including nuclear receptor activation, limiting endoplasmic reticulum stress, and blocking mitochondrial dysfunction. In animal models of inflammatory pain and diabetic neuropathic pain, small molecule inhibitors of soluble epoxide hydrolases have shown strong analgesic effects (Neurotherapeutics, 2020, 17, 900-916). EETs are readily metabolized and inactivated by soluble epoxide hydrolase (sEH) in vivo, and the EET metabolite dihydroxy DHETs has inflammatory effects. Small molecule inhibitors of soluble epoxide hydrolase can stabilize EpFA in vivo. Therefore, increasing the amount of EETs in the body by inhibiting sEH activity has become a new approach to treating EETs-related diseases.

[0006] EpFA exerts its analgesic effects through multiple mechanisms, such as reducing endoplasmic reticulum (ER) stress, preventing or reversing endothelial cell dysfunction (ECD), and stabilizing mitochondrial function (Cell Physiol Biochem, 2015, 36, 474-486). EpFA can modulate cellular stress induced by reactive oxygen species (ROS) and shift the ER stress response to homeostasis rather than activating inflammatory pathways that lead to cellular senescence and death. EpFA can reduce the ER stress response and limit ROS, indirectly maintaining mitochondrial functional stability. EpFA can also directly block the effects of mitochondrial dysfunction. Inhibition of sEH activity can stabilize EpFA and limit the production of some pro-inflammatory diol metabolites. Therefore, EpFA mediates beneficial effects in all these processes, shifting the ER stress response to homeostasis and alleviating pain.

[0007] There is substantial evidence that EpFA plays a role in nociception by blocking inflammatory and neuropathic pain, thus sEH inhibitors and EpFA analogues have great potential in alleviating human pain.

[0008] Epidemiological studies have confirmed the association of inflammatory biomarkers with the development of type 2 diabetes mellitus (T2DM) and its complications. The triggering mechanisms of inflammation in T2DM remain unclear. Inflammatory responses may contribute to T2DM by inducing insulin resistance, with an obese environment leading to adipose tissue dysfunction, macrophage infiltration, and increased release of cytokines such as IL-6 and TNF-α. Long-term elevated levels of these molecules promote insulin resistance in skeletal muscle and endothelial dysfunction in the vascular system, as well as the release of acute-phase proteins from the liver. Chronicly elevated levels of specific inflammatory markers, such as IL-6 and TNF-α, appear to be associated with metabolic disorders and can alter insulin sensitivity by triggering different key steps in the insulin signaling pathway. Hyperglycemia can also induce IL-6 production in endothelial cells and macrophages. Furthermore, hyperglycemia enhances the effects of cytokine signaling inhibitors (SOCSs), thereby impairing insulin release and signal transduction, promoting long-term complications of diabetes. Targeting inflammatory pathways may be an integral part of strategies for the prevention and control of diabetes and its related complications.

[0009] PPAR agonists have been shown to inhibit the expression of cytokines—such as resistin, tumor necrosis factor-α (TNFα), and interleukin-6—that promote insulin resistance. PPAR agonists induce an increase in plasma adiponectin concentrations, a hormone secreted from adipose tissue and present in low levels in the plasma of patients with type 2 diabetes mellitus (T2DM). Adiponectin increases fatty acid oxidation in the liver and skeletal muscle. Overall, adiponectin enhances insulin sensitivity in skeletal muscle and liver and reduces glucose production in the liver, thereby decreasing circulating FFA and TG levels, as well as glucose levels. Macrophage infiltration in obese adipose tissue is involved in local inflammation that enhances insulin resistance. Recent studies have shown that the PPAR component in macrophages mediates the antidiabetic effects of TZD. Inactivation of PPARs in macrophages leads to impaired activation of alternative macrophages, glucose intolerance, and insulin resistance in skeletal muscle and liver.

[0010] Currently, the main analgesics used in clinical practice include opioid analgesics and nonsteroidal anti-inflammatory drugs (NSAIDs). Both have certain side effects. For example, traditional opioid analgesics have a strong effect, but they also have strong addictive properties and side effects such as respiratory depression, low blood pressure, nausea, vomiting, constipation, and difficulty urinating. NSAIDs are divided into non-selective NSAIDs and selective cyclooxygenase-2 (COX-2) inhibitors. Although they also have good analgesic effects, non-selective NSAIDs have severe gastrointestinal irritation and can easily lead to gastric ulcers. They also often have adverse reactions on the coagulation and hematopoietic systems. Although selective COX-2 inhibitors do not have gastrointestinal irritation, they can easily cause an imbalance of prostacyclin and thromboxane, leading to cardiovascular diseases, and generally have little effect on neuropathic pain.

[0011] Currently, the main hypoglycemic drugs used in clinical practice include insulin, insulin secretagogues, insulin sensitizers, alpha-glucosidase inhibitors, GLP-1 agonists, and dipeptidyl peptidase-4 inhibitors, all of which have certain drawbacks. For example, insulin alpha-glucosidase inhibitors cannot be used alone to treat diabetes and usually need to be combined with other drugs for blood sugar control; insulin secretagogues can cause gastrointestinal symptoms (such as nausea and upper abdominal distension) and headaches; insulin must be injected for treatment, resulting in poor patient compliance; and other drugs are difficult to combat diabetic inflammatory complications and neuropathic pain.

[0012] Currently reported dual-target compounds for sEH inhibitors and PPARs agonists include RB394 and sEH IC. 50 =0.3μM, PPARγEC 50 =0.3μM, it has shown some efficacy in the treatment of diabetic nephropathy and steatohepatitis, but further development and exploration are still needed. Given that dual-target compounds of sEH inhibitors and PPARs agonists can not only lower blood glucose but also effectively treat diabetic inflammatory complications, non-alcoholic fatty liver disease, and neuropathic pain, reducing drug interactions that previously required multiple medications, the development of novel and more effective dual-target compounds of sEH inhibitors and PPARs agonists for the treatment of diabetes, non-alcoholic fatty liver disease, pain, and depression is urgent and necessary. Summary of the Invention

[0013] The purpose of this invention is to provide a compound, its preparation method, and its application in the preparation of sEH inhibitors and PPARs agonists. The compound provided by this invention has high activity against human sEH (HsEH) and PPARs, with few side effects, and can be used as a dual-target compound for the treatment of diabetes and its inflammatory complications, neuropathic pain, and depression.

[0014] This invention provides a compound having the structure shown in formula V or VI:

[0015]

[0016] Wherein, R1 is arginyl, aryl, alkyl-substituted aryl, haloaryl, haloalkyl-substituted aryl, haloalkoxy-substituted aryl, haloaryloxy-substituted aryl; R4 is hydrogen or alkyl; R6 is hydrogen, hydroxyl, alkyl, ester or amino; A is cycloalkyl, heterocyclic or aryl; B is single bond, cycloalkyl, heterocyclic or aryl;

[0017] W is a single bond, -CH2-, -O-, -S-, -NH-, or

[0018] Y is a single bond, -CH2-, -O-, -S-, or -NH-; Z is =CH2, =O, =S, or =NH; n is an integer from 0 to 12.

[0019] This invention provides a compound having the structure shown in Formula V or VI. The compound provided by this invention has a typical urea structure as the primary pharmacophore of soluble epoxide hydrolase (sEH), and a thiazolidinedione moiety as the primary pharmacophore of peroxisome proliferator-activated receptors (PPARs). The sEH inhibitor and PPARs agonist compound provided by this invention exhibits high inhibitory activity against human HsEH and high agonistic activity against PPARs, and can be used as an sEH inhibitor and PPARs agonist compound in the preparation of drugs for treating diseases mediated by soluble epoxide hydrolase and peroxisome proliferator-activated receptors. Attached Figure Description

[0020] Figure 1 This is a reaction route diagram of the sEH inhibitor having the structure shown in Formula V and the PPARs agonist compound in this invention.

[0021] Figure 2 The effect of compound SP-CO1 on the body weight of KM mice was prepared as an example.

[0022] Figure 3 The effect of compound SP-CO1 on the visceral index of KM mice was prepared as an example.

[0023] Figure 4 Representative H&E stained sections of compound SP-C01 on the effects of KM mouse organs were prepared for this example.

[0024] Figure 5 To evaluate the efficacy of compound SP-C01 in treating CFA-induced arthritis in a mouse model (AIA);

[0025] Figure 6 To evaluate the efficacy of compound SP-C01 in treating CFA-induced arthritis in a mouse model (AIA);

[0026] Figure 7 In a chronic contractile injury model of neuropathy in male SD rats, SP-C01 blocked pain as measured by the von Frey test (mechanical withdrawal threshold);

[0027] Figure 8 Histological analysis of pancreatic tissue in mice treated with control, Mod, celecoxib, ulinastatin, and compound SP-C01. Representative H&E stained sections of pancreas from the in vivo efficacy studies;

[0028] Figure 9The results of histological analysis of pancreatic tissue in mice treated with control, Mod, celecoxib, ulinastatin and compound SP-C01;

[0029] Figure 10 The results show the blood glucose analysis of mice treated with control, Mod, pioglitazone, compound SP-B07, and compound SP-C01.

[0030] Figure 11 The results of analysis on diabetic neuropathic pain in mice treated with control, Mod, pioglitazone, compound SP-B07 and compound SP-C01;

[0031] Figure 12 This is a reaction route diagram of the sEH inhibitor having the structure shown in Formula VI and the PPARs agonist compound in this invention.

[0032] Figure 13 This is a reaction route diagram of the sEH inhibitor with the structure shown in Formula V when Y is -NH- in this invention, and the PPARs agonist compound. Detailed Implementation

[0033] This invention provides a compound having the structure shown in formula V or VI:

[0034]

[0035] Wherein, R1 is adamantyl, aryl, alkyl-substituted aryl, haloaryl, haloalkyl-substituted aryl, or haloalkoxy-substituted aryl; R4 is hydrogen or alkyl; R6 is hydrogen, hydroxyl, alkyl, ester or amino; A is cycloalkyl, heterocyclic or aryl; B is a single bond, cycloalkyl, heterocyclic or aryl.

[0036] W is a single bond, -CH2-, -O-, -S-, -NH-, or

[0037] Y is a single bond, -CH2-, -O-, -S-, or -NH-; Z is independently =CH2, =O, =S, or =NH; n is an integer from 0 to 12.

[0038] In this invention, the alkyl group in the alkyl-substituted aryl group and the alkyl-substituted aryl group are preferably methyl, ethyl, propyl, butyl, pentyl, isobutyl, isopropyl, isopentyl, or tert-butyl; the alkoxy group in the alkoxy-substituted aryl group is preferably methoxy, ethoxy, propoxy, isopropoxy, butoxy, cyclopentoxy, cyclohexyloxy, phenoxy, or benzyloxy; and the halogen in the aryl group, the alkyl-substituted aryl group, and the alkoxy-substituted aryl group is preferably -F, -Cl, or -Br.

[0039] In this invention, the cycloalkyl group is an unsubstituted or substituted C3-C8 cycloalkyl group; the substituent of the substituted C3-C8 cycloalkyl group is preferably -F, -Cl, -Br, -OH, -NH2, -NHCH3, -N(CH3)2 or C1-C6 alkyl; the heterocyclic group is preferably an unsubstituted or substituted 3- to 10-membered heterocyclic group; the substituent of the substituted 3- to 10-membered heterocyclic group is preferably -F, -Cl, -Br, -OH, -NH2, -NHCH3, -N(CH3)2 or C1-C6 alkyl; the aryl group is preferably a substituted or unsubstituted phenyl, pyridyl or naphthyl group; the substituent of the substituted phenyl, pyridyl or naphthyl group is preferably -F, -Cl, -Br, -OH, -NH2, -NHCH3, -N(CH3)2 or C1-C6 alkyl.

[0040] In this invention, R1 is preferably ammonium alkyl, haloaryl, haloalkyl-substituted aryl, or haloalkoxy-substituted aryl; R4 is preferably hydrogen, methyl, or ethyl; R6 is preferably hydroxyl, amino, alkyl, or ester; A is preferably cyclohexyl or phenyl; B is preferably a single bond or phenyl; W is preferably a single bond or -O-; Y is preferably a single bond, -O-, or -NH-; Z is =O; n is preferably an integer from 0 to 2.

[0041] In this invention, R1 is preferably adamantyl alkyl group. R4 is preferably hydrogen; R6 is preferably hydroxyl, -NH2, methyl, -C(O)-O-CH2-CH3 or sec-butyl.

[0042] In this invention, the compound preferably has any one of the following structures:

[0043]

[0044] This invention provides a method for preparing the compound described in the above technical solution.

[0045] (5) Prepare a compound having the structure shown in Formula V. The preparation method includes the following steps:

[0046] Compound a and compound h were subjected to a first substitution reaction to obtain compound i;

[0047] The compound i was subjected to a first hydrolysis reaction to obtain compound v;

[0048] Compounds v and w were subjected to a first nucleophilic substitution reaction with compound af to obtain a compound having the structure shown in formula V;

[0049] The structural formulas of compounds a, h, i, and v are as follows:

[0050]

[0051] In this case, Q in chemical a is H, hydroxyl, amino, thiol, carboxyl, or acyl chloride;

[0052] The structural formulas of compounds w and af are as follows:

[0053]

[0054] (6) Prepare a compound having the structure shown in Formula VI, the preparation method comprising the following steps:

[0055] Compound a and compound x undergo a first substitution reaction to obtain compound y;

[0056] The compound y was subjected to a first hydrolysis reaction to obtain compound z;

[0057] Compound z, compound w and compound af were subjected to a first nucleophilic substitution reaction to obtain a compound having the structure shown in formula V;

[0058] The structural formulas of compounds a, x, y, and z are as follows:

[0059]

[0060] In this compound, Q in chemical a is H, hydroxyl, amino, mercapto, carboxyl, or acyl chloride; X in compound x is H, hydroxyl, halogen, or haloalkyl.

[0061] The structural formulas of compounds w and af are as follows:

[0062]

[0063] (7) When Y in the compound with the structure shown in formula V or VI is -NH-, the method for preparing the compound having the structure shown in formula V or VI may include the following steps:

[0064] Compound a undergoes a first substitution reaction with compound ab to obtain compound ac;

[0065] The compound ac was subjected to a first hydrolysis reaction to obtain compound ad;

[0066] Compounds ad and w were subjected to a nucleophilic substitution reaction with compound af to obtain compound ae;

[0067] The compound ae was subjected to a first reduction reaction to obtain a compound having the structure shown in Formula VI;

[0068] A compound having the structure shown in Formula VI is subjected to a condensation reaction with compound ag to obtain a compound having the structure shown in Formula V;

[0069]

[0070] In this compound, Q in compound a is H, hydroxyl, amino, mercapto, carboxyl, or acyl chloride; X in compound ab is H, hydroxyl, halogen, or haloalkyl.

[0071] In this invention, unless otherwise specified, all raw materials / components used in the preparation are commercially available products well known to those skilled in the art.

[0072] like Figure 1 The reaction route shown is as follows: In this invention, compound a undergoes a nucleophilic substitution reaction with compound h to obtain compound i. In this invention, the nucleophilic substitution reaction is preferably carried out in the presence of triphenylphosphine and DIAD. In this invention, the molar ratio of compound a, compound h, triphenylphosphine, and DIAD is preferably 1:1:1.5:1.5. In this invention, the solvent for the nucleophilic substitution reaction is preferably tetrahydrofuran, the nucleophilic substitution reaction is preferably carried out under ice-salt bath conditions, and the reaction time is preferably 8-12 hours. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0073] After obtaining compound i, compound i is subjected to a deprotection reaction under acidic conditions to obtain v. In this invention, the reagent providing the acidic conditions is preferably trifluoroacetic acid, and the deprotection reaction is preferably carried out under dichloromethane conditions. In this invention, the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 1.5 to 2.5 h. After the deprotection reaction, the resulting reaction solution is preferably subjected to vacuum distillation, water and dichloromethane are added to the residue, the pH of the system is adjusted to 14 with sodium hydroxide solid under ice-water bath conditions, the organic layer is separated and removed, the aqueous layer is extracted with dichloromethane (100 mL × 2), and then washed successively with water, saturated brine, and dried with anhydrous sodium sulfate, filtered, and the resulting filtrate is concentrated under vacuum to obtain compound v.

[0074] After obtaining compound v, the present invention performs a first nucleophilic substitution reaction on compound v, compound w, and compound af to obtain an sEH inhibitor and PPARs agonist compound having the structure shown in formula V. In the present invention, the molar ratio of compound v to compound af is preferably 1:(0.33–0.6), more preferably 1:(0.4–0.5). In the present invention, the nucleophilic substitution reaction is preferably carried out in the presence of triethylamine. In the present invention, the nucleophilic substitution reaction is preferably carried out under ice bath conditions, and the reaction time is preferably 10–50 min, more preferably 30 min. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0075] like Figure 12 The reaction route shown is as follows: In this invention, compound a undergoes a nucleophilic substitution reaction with compound x to obtain compound y. In this invention, the nucleophilic substitution reaction is preferably carried out in the presence of triphenylphosphine and DIAD. In this invention, the molar ratio of compound a, compound x, triphenylphosphine, and DIAD is preferably 1:1:1.5:1.5. In this invention, the solvent for the nucleophilic substitution reaction is preferably tetrahydrofuran, the nucleophilic substitution reaction is preferably carried out under ice-salt bath conditions, and the reaction time is preferably 8-12 hours. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0076] After obtaining compound y, compound y is subjected to a deprotection reaction under acidic conditions to obtain z. In this invention, the reagent providing the acidic conditions is preferably trifluoroacetic acid, and the deprotection reaction is preferably carried out under dichloromethane conditions. In this invention, the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 1.5 to 2.5 h. After the deprotection reaction, the resulting reaction solution is preferably subjected to vacuum distillation, water and dichloromethane are added to the residue, the pH of the system is adjusted to 14 with sodium hydroxide solid under ice-water bath conditions, the organic layer is separated and removed, the aqueous layer is extracted with dichloromethane (100 mL × 2), and then washed successively with water, saturated brine, and dried with anhydrous sodium sulfate, filtered, and the resulting filtrate is concentrated under vacuum to obtain compound z.

[0077] After obtaining compound z, the present invention performs a first nucleophilic substitution reaction on compound z, compound w, and compound af to obtain an sEH inhibitor and PPARs agonist compound having the structure shown in Formula VI. In the present invention, the molar ratio of compound z to compound af is preferably 1:(0.33–0.6), more preferably 1:(0.4–0.5). In the present invention, the nucleophilic substitution reaction is preferably carried out in the presence of triethylamine. In the present invention, the nucleophilic substitution reaction is preferably carried out under ice bath conditions, and the reaction time is preferably 10–50 min, more preferably 30 min. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0078] like Figure 13The reaction route shown is as follows: In this invention, compound a undergoes a first nucleophilic substitution reaction with compound ab to obtain compound ac. In this invention, the nucleophilic substitution reaction is preferably carried out in the presence of triphenylphosphine and DIAD. In this invention, the molar ratio of compound a, compound ab, triphenylphosphine, and DIAD is preferably 1:1:1.5:1.5. In this invention, the solvent for the nucleophilic substitution reaction is preferably tetrahydrofuran, the nucleophilic substitution reaction is preferably carried out under ice-salt bath conditions, and the reaction time is preferably 8-12 hours. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0079] After obtaining compound ac, compound ac undergoes a first hydrolysis reaction to obtain compound ad. The first hydrolysis reaction is a deprotection reaction under acidic conditions to obtain ad. In this invention, the reagent providing the acidic conditions is preferably trifluoroacetic acid, and the deprotection reaction is preferably carried out under dichloromethane conditions. In this invention, the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 1.5 to 2.5 hours. After the deprotection reaction, the resulting reaction solution is preferably subjected to vacuum distillation. Water and dichloromethane are added to the residue, and the pH of the system is adjusted to 14 with solid sodium hydroxide under ice-water bath conditions. The organic layer is separated and removed, and the aqueous layer is extracted with dichloromethane (100 mL × 2). Then, it is washed successively with water, saturated brine, and dried with anhydrous sodium sulfate. After filtration, the filtrate is concentrated under vacuum to obtain compound ad.

[0080] After obtaining compound ad, the present invention involves reacting compound ad, compound w, and compound af with a nucleophilic substitution reaction to obtain compound ae. In the present invention, the molar ratio of compound ad to compound af is preferably 1:(0.33–0.6), more preferably 1:(0.4–0.5). In the present invention, the nucleophilic substitution reaction is preferably carried out in the presence of triethylamine. In the present invention, the nucleophilic substitution reaction is preferably carried out under ice bath conditions, and the reaction time is preferably 10–50 min, more preferably 30 min. After the nucleophilic substitution reaction, the mixture is purified by column chromatography.

[0081] After obtaining compound ae, the present invention performs a first reduction reaction on compound ae to obtain a compound having the structure shown in formula VI. In the present invention, the reducing agent for the first reduction reaction is preferably hydrogen, the catalyst is Pb-C, and the solvent is methanol. In the present invention, the mass ratio of compound ae to Pd-C is 1:0.1. The reduction reaction is preferably carried out under reflux conditions, and the reduction reaction time is preferably 4 to 8 hours. After the reduction reaction, the reaction solution is filtered and evaporated to dryness.

[0082] After obtaining the compound having the structure shown in Formula VI, the present invention performs a condensation reaction between the compound having the structure shown in Formula VI and compound ag to obtain the compound having the structure shown in Formula V. In the present invention, the condensing agent is preferably HATU, the base is preferably DIEA, and the solvent is preferably THF. In the present invention, the condensation reaction temperature is preferably room temperature, and the time is preferably 4–8 h. The molar ratio of the compound having Formula VI to HATU and DIEA is preferably 1:1.5:3. After the condensation reaction, the present invention preferably performs vacuum distillation on the obtained reaction solution, adds water and dichloromethane to the obtained residue for extraction, washes with 1N HCl, and dries the organic layer with anhydrous sodium sulfate. The residue is filtered, concentrated to dryness under reduced pressure, and purified by column chromatography to obtain the compound having the structure shown in Formula V.

[0083] This invention provides the application of the aforementioned compounds and their pharmaceutically acceptable deuterated derivatives, salts or hydrates, or compounds prepared by the preparation methods described above, in the preparation of peroxisome proliferator-activated receptor agonists and soluble epoxide hydrolase inhibitors.

[0084] In this invention, the peroxisome proliferator-activated receptor agonist and the soluble epoxide hydrolase inhibitor are used to treat soluble epoxides and diseases mediated by peroxisome proliferator-activated receptors; the soluble epoxides and diseases mediated by peroxisome proliferator-activated receptors preferably include inflammatory diseases, pain, sepsis, cardiovascular diseases, neurodegenerative diseases, diabetes, diabetic complications, depression, liver fibrosis, renal failure, chronic obstructive pulmonary disease, or pulmonary hypertension.

[0085] In this invention, the inflammatory disease preferably includes non-alcoholic steatohepatitis or chronic nephritis; the pain includes neuropathic pain.

[0086] This invention provides a sEH inhibitor and PPARs agonist compound. The sEH inhibitor and PPARs agonist compound provided by this invention has a typical urea structure as the primary pharmacophore of sEH and a thiazolidinedione moiety as the primary pharmacophore of PPARs. The sEH inhibitor and PPARs agonist compound provided by this invention exhibits high inhibitory activity against human HsEH and high agonistic activity against PPARs, and can be used as sEH inhibitors and PPARs agonists in the preparation of drugs for treating diseases mediated by soluble cyclooxygenase and peroxisome proliferator-activated receptors.

[0087] This invention provides an sEH inhibitor and a PPARs agonist compound, or a pharmaceutically acceptable combination thereof, as well as their preparation method and application, belonging to the field of pharmaceutical technology. The sEH inhibitor and PPARs agonist compound provided by this invention has the structure shown in Formula I, II, III, IV, V, or VI. The sEH inhibitor portion provided by this invention can stabilize endogenous epoxidized fatty acids with broad physiological activity, exhibiting a strong inhibitory effect on recombinant human sEH. It can act through multiple mechanisms, including regulating the production of various pro-inflammatory cytokines, reducing endoplasmic reticulum stress, preventing or reversing endothelial dysfunction, and stabilizing mitochondrial function. The PPAR agonist can synergistically inhibit the expression of cytokines—such as resistin, tumor necrosis factor α (TNFα), and interleukin-6—which promote insulin resistance. The PPAR agonist induces an increase in plasma adiponectin concentration. Adiponectin is a hormone secreted from adipose tissue and is present in low levels in the plasma of patients with type 2 diabetes. Adiponectin increases fatty acid oxidation in the liver and skeletal muscle. Therefore, the compounds involved in this invention can be used for soluble cyclooxide enzymes and peroxisome proliferator-activated receptor-mediated diseases including inflammatory diseases, pain, sepsis, cardiovascular diseases, neurodegenerative diseases, diabetes, diabetic complications, depression, non-alcoholic steatohepatitis, liver fibrosis, chronic nephritis, renal failure, chronic obstructive pulmonary disease, or pulmonary hypertension.

[0088] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0089] Example 1: Synthesis of (1r,4r)-4-hydroxycyclohexyl)tert-butyl carbamate

[0090] Add trans-4-aminocyclohexanol hydrochloride (5.00 g, 32.98 mmol), sodium carbonate (10.49 g, 98.98 mmol), and water (25 mL) sequentially to a 100 mL three-necked flask. Cool to 0 °C in an ice bath, then add 5 mL of DCM solution containing Boc₂O (7.92 g, 36.28 mmol) dropwise over 3 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. After 4 hours, monitor with TLC (EA:PE = 1:3, phosphomolybdic acid colorimetric indicator) to confirm the reaction has occurred, and stop the reaction. Pour the reaction solution into 50 mL of water, extract with DCM (30 mL × 3), extract with EA (30 mL × 3), combine the organic layers, wash with water (30 mL), wash with saturated NaCl (30 mL), and dry to anhydrous magnesium sulfate. Filter under vacuum, wash with EA, and concentrate the filtrate to dryness under reduced pressure to obtain 8.0 g of crude white powder. The crude product was directly added to the next step without purification.

[0091] Example 2 Synthesis of 4-aminophenyl-4-nitrobenzene ester

[0092] To a 100 mL three-necked flask, add cis-4-Boc-aminocyclohexanol (7.10 g, 32.98 mmol), p-nitrobenzoic acid (5.51 g, 98.98 mmol), triphenylphosphine (12.98 g, 49.47 mol), and THF (20 mL). Cool to below -10 °C in an ice-salt bath, and then add DIAD (10 g, 49.47 mol) in THF solution (20 mL). After 2 hours, monitor the reaction by TLC. Once the reaction is complete, remove the solvent under reduced pressure to obtain 35.6 g of crude product. Purify the crude product by silica gel column chromatography. Mix 1.5 times the sample volume, pack 5 times the silica gel volume into a column, and use EA:PE = 1:10 as the eluent to obtain 5.53 g of white solid.

[0093] Example 3: tert-butyl (4-hydroxyphenyl) carbamate

[0094] 4-Aminophenyl-4-nitrobenzene ester (5.53 g, 14.88 mmol), NaOH (1.79 g, 44.65 mmol), THF (20 mL), and H2O (20 mL) were added sequentially to a 100 mL three-necked flask. The reaction was carried out at room temperature. After 4 h, the reaction was monitored by TLC. After the reaction was completed, the mixture was concentrated to dryness under reduced pressure. EA (30 mL) and saturated sodium carbonate (20 mL × 2) were added for extraction. The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to give 3.01 g of white solid, with a yield of 94.08%.

[0095] Example 4: tert-butyl ((1r,4r)-4-(4-formylphenoxy)cyclohexyl)carbamate

[0096] Add cis-N-BOC-4-aminocyclohexanol (2.50 g, 11.61 mmol, 1 eq), p-hydroxybenzaldehyde (1.42 g, 11.61 mmol, 1 eq), triphenylphosphine (4.57 g, 17.42 mmol, 1.5 eq), and THF (10 mL) sequentially to a 500 mL three-necked flask. Cool the flask to below -10 °C in an ice-salt bath. Add 5 mL of a THF solution containing DIAD (3.52 g, 17.42 mmol, 1.5 eq) dropwise at a rate of one drop every two seconds. After 72 hours, TLC monitoring showed that EA:PE = 1:1, indicating that the reaction was largely complete; therefore, the reaction was stopped. THF was removed by vacuum concentration, yielding 12.21 g of brownish-yellow oily substance. The crude product was purified by silica gel column chromatography with 1.5 times the sample volume mixed and packed into a 10-fold silica gel column. The eluent was EA:PE = 1:30, yielding 2.85 g of white solid, with a yield of 76.82%.

[0097] Example 5: Synthesis of tert-butyl ((1r,4r)-4-{4-[(Z)-(2,4-dioxothiazolidin-5-ylidene)methyl]phenoxy}cyclohexyl)carbamate

[0098] To a 100 mL round-bottom flask, tert-butyl[(1r,4r)-4-(4-formylphenoxy)cyclohexyl]carbamate (2.85 g, 8.93 mmol, 1 eq), 2,4-thiazolidinedione (1.05 g, 8.93 mmol, 1 eq), pyridine (0.35 g, 4.46 mmol, 0.5 eq), glacial acetic acid (0.27 g, 4.46 mmol, 0.5 eq), and toluene (10 mL) were added sequentially, and the mixture was heated to reflux. A solid precipitated as the reaction proceeded. After 8 hours, TLC monitoring indicated the reaction was complete, and the reaction was stopped. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with a small amount of toluene to give 2.03 g of a white solid. ESI-MS (m / z): 424.1 [M+H] +

[0099] Example 6 Synthesis of 5-{(Z)-4-[(1r,4r)-4-aminocyclohexyl]oxybenzyl}thiazolidin-2,4-dione

[0100] To a 500 mL three-necked flask, tert-butyl((1r,4r)-4-{4-[(Z)-(2,4-dioxothiazolidin-5-ylidene)methyl]phenoxy}cyclohexyl)carbamate (2.03 g, 4.85 mmol), DCM (15 mL), and the mixture was cooled to 0 °C in an ice bath before adding TFA (4 mL) dropwise. After 0.5 hours, TLC monitoring showed that the reaction was complete, and the reaction was stopped. TFA was removed from the reaction mixture by vacuum distillation. Water (30 mL) was added, and the mixture was extracted with DCM (30 mL × 3). The organic layers were combined and concentrated to dryness under reduced pressure. 1.17 g of crude brown solid was obtained. The crude product was used directly in the next step without purification.

[0101] Example 7 Synthesis of 4-(4-formylphenoxy)piperidine-1-carboxylic acid tert-butyl ester

[0102] N-Boc-4-hydroxypiperidine (5.00 g, 0.25 mol), p-hydroxybenzaldehyde (15.18 g, 0.25 mol), triphenylphosphine (48.92 g, 0.37 mol), and THF (75 mL) were added sequentially to a 500 mL three-necked flask. The flask was cooled to below -10 °C in an ice-salt bath, and a THF solution (80 mL) of DIAD (37.74 mg, 0.37 mol) was added dropwise at a rate of one drop every two seconds. After 5 hours, the reaction was monitored by TLC and found to be complete. The reaction was then stopped. The THF was removed by concentration under reduced pressure, yielding 127.01 g of a brownish-yellow oily substance. The crude product was used directly for the next step without purification. 1H NMR (400MHz, CDCl3): δ (ppm) 9.88 (s, 1H), 7.82 (d, J = 2.0Hz, 2H), 7.01 (d, J = 2.0Hz, 2H), 4.64-4.59 (m,1H),3.73-3.67(m,2H),3.42-3.35(m,2H),1.99-1.92(m,2H),1.82-1.74(m,2H),1.77(s,9H).

[0103] Example 8 Synthesis of 4-(piperidin-4-oxy)benzaldehyde

[0104] 127.01 g of 4-(4-formylphenoxy)piperidine-1-carboxylic acid tert-butyl ester and 180 mL of DCM were added sequentially to a 500 mL three-necked flask. The mixture was cooled to 0 °C in an ice bath, and TFA (119.04 g, 1.04 mol) was added dropwise. After 37 hours, the reaction was monitored by TLC and found to be complete, at which point the reaction was stopped. The TFA in the reaction solution was removed by vacuum distillation. 50 mL of DCM was added, and the mixture was extracted with 1 N HCl (200 mL × 6). The aqueous layers were combined, and the pH was adjusted to 10 with NaOH solid. The mixture was then extracted with n-butanol (200 mL × 12). The organic layers were combined, concentrated to dryness under reduced pressure, and slurried with 12 mL of acetone. The mixture was filtered, and the filter cake was washed with 2 mL of acetone and dried to give 17.45 g of crude product, a brownish-red solid. The combined yield of the two steps was 34%. The crude product was used directly in the next step without purification.

[0105] Example 9 Synthesis of (Z)-5-[4-(piperidin-4-oxy)benzylene]thiazolidin-2,4-dione

[0106] Add 4-(piperidin-4-oxy)benzaldehyde (4.00 g, 19.50 mmol), 2,4-thiazolidinedione (2.28 g, 19.50 mmol), pyridine (0.77 g, 9.75 mmol), glacial acetic acid (0.59 g, 9.75 mmol), and toluene (15 mL) sequentially to a 250 mL round-bottom flask, and heat to reflux. A solid precipitated as the reaction proceeded. After 8 hours, TLC monitoring showed the reaction was complete, and the reaction was stopped. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with a small amount of toluene to obtain 3 g of a pale yellow solid. ESIMS: m / z 304.10 [M+H] + .

[0107] Example 10 Synthesis of (Z)-5-[4-(piperidin-4-oxy)benzylidene]imidazoline-2,4-dione

[0108] Add 1.95 g (19.50 mmol) of 2,4-imidazolidinedione and 5 mL of water to a 100 mL round-bottom flask. Heat to 70 °C, adjust the pH to 7 with saturated sodium bicarbonate solution, add ethanolamine (2.38 g, 19.50 mmol), heat to 100 °C, and add dropwise 15 mL of EtOH solution containing 4-(piperidin-4-oxy)benzaldehyde (4 g, 19.50 mmol). A solid precipitates as the reaction proceeds. After 16 hours, TLC monitoring indicates the reaction is complete, and the reaction is stopped. Cool the reaction solution to room temperature, filter, and wash the filter cake with a small amount of water to obtain 1.8 g of white solid. ESIMS: m / z 287.13 [M+H] + .

[0109] Example 11 Synthesis of 4-{(1r,4r)-4-[(tert-butoxycarbonyl)amino]cyclohexyl}oxy)phenylacetic acid ester

[0110] To a 500 mL three-necked flask, trans-4-Boc-aminocyclohexanol (9.27 g, 43.08 mmol, 1 eq), 4-hydroxyphenylacetic acid ester (6.55 g, 43.08 mmol, 1 eq), triphenylphosphine (16.94 g, 64.62 mmol, 1.5 eq), and THF (25 mL) were added sequentially. The flask was cooled to below -10 °C in an ice-salt bath. A THF solution of DIAD (13.07 g, 64.62 mmol, 1.5 eq) (12.5 mL) was then added dropwise at a rate of one drop every two seconds. After 72 hours, TLC monitoring showed the reaction was complete, and the reaction was stopped. The THF was removed by concentration under reduced pressure, yielding 35.57 g of a brownish-yellow oily substance. The crude product was purified by silica gel column chromatography. The column was packed with 10x silica gel using 1.5x stirred sample and EA:PE = 1:30 as eluent, yielding 2.33 g of a white solid.

[0111] Example 12 Synthesis of tert-butyl[(1r,4r)-4-(4-hydroxyphenoxy)cyclohexyl]carbamate

[0112] Add 4-{(1r,4r)-4-[(tert-butyloxycarbonyl)amino]cyclohexyl}oxy)phenylacetate (0.16 g, 0.46 mmol, 1 eq), THF (5 mL), H2O (0.5 mL), and LiOH (0.03 g, 1.37 mmol, 3 eq) sequentially to a 25 mL single-necked flask and stir at room temperature. After 50 min, monitor with TLC, stop the reaction, concentrate under reduced pressure to remove THF, extract with EA (30 mL), adjust pH to 4 with 1 N HCl, dry the organic layer with anhydrous magnesium sulfate (30 mL × 3), filter, concentrate under reduced pressure to dryness, and give 0.10 g of crude pale yellow solid. ESIMS: m / z 330.2 [M+H] + .

[0113] Example 13 Synthesis of methyl 2-(4-{(1r,4r)-4-[(tert-butoxycarbonyl)amino]cyclohexyl)oxy}phenoxy)acetate

[0114] To a 25 mL single-necked flask, tert-butyl[(1r,4r)-4-(4-hydroxyphenoxy)cyclohexyl]carbamate (0.43 g, 1.40 mmol, 1 eq), MeCN (15 mL), K₂CO₃ (0.58 g, 4.20 mmol, 3 eq), and KI (0.023 g, 0.14 mmol, 0.1 eq) were added dropwise. Methyl bromoacetate (0.32 g, 2.10 mmol, 1.5 eq) was added dropwise, followed by Ar displacement three times. The mixture was heated to reflux. After 16 hours, the reaction was stopped by TLC monitoring. The mixture was concentrated to dryness under reduced pressure, and extracted with water (30 mL), followed by DCM extraction (30 mL × 3). The organic layers were combined, washed with water (30 mL), washed with saturated brine (30 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain 0.78 g of a brownish-black oily liquid. The crude product was purified by silica gel column chromatography. The column was packed with 1.5 times the volume of sample and 3 times the volume of silica gel. The eluent was EA:PE = 1:5, yielding 0.40 g of a white solid, with a yield of 75.35%. ESIMS: m / z 402.1 [M+H]+.

[0115] Example 14 Synthesis of methyl 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)acetate

[0116] 2-(4-{(1r,4r)-4-[(tert-butoxycarbonyl)amino]cyclohexyl)oxy}phenoxy)methyl acetate (0.30 g, 0.79 mmol), and DCM (10 mL) were added sequentially to a 25 mL single-necked flask. The mixture was cooled to 0 °C in an ice bath, and TFA (3 mL) was added dropwise. After 2 hours, the reaction was monitored by TLC and found to be complete. The reaction was then stopped. The TFA in the reaction solution was removed by vacuum distillation, yielding 0.21 g of crude brown oil.

[0117] Example 15 Synthesis of 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)methyl acetate (SP-CO3)

[0118] Add BTC (0.08 g, 0.27 mmol, 0.34 eq), Et3N (0.16 g, 1.58 mmol, 3 eq), and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C in a cold trap. Add a 5 mL solution of 3-fluoro-4-trifluoromethoxyaniline (0.21 g, 0.79 mmol, 1 eq) in DCM dropwise over 3 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0119] A 10 mL solution of 3-fluoro-4-trifluoromethoxyaniline isocyanate in DCM and Et3N (0.16 g, 1.58 mmol, 3 eq) was added dropwise to a 25 mL three-necked flask. The mixture was kept on ice at 0 °C, and a 10 mL solution of 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)methyl acetate (0.22 g, 0.79 mmol, 1 eq) in DCM was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed the reaction was complete after 30 min, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 0.44 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 10x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:5, yielding 0.10 g of white solid (yield 25.6%). ESIMS: m / z 523.0 [M+Na] + .

[0120] Example 16 Synthesis of 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)acetic acid (SP-CO2)

[0121] To a 25 mL single-necked flask, methyl 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)acetate (70 mg, 0.14 mmol, 1 eq), THF (2 mL), H₂O (0.5 mL), and LiOH (10 mg, 0.42 mmol, 3 eq) were added sequentially, and the mixture was stirred at room temperature. After 50 min, the reaction was stopped by TLC monitoring. The mixture was concentrated under reduced pressure to remove THF, and EA (30 mL) was added. The mixture was extracted with water (20 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to obtain 20.02 mg of a white solid. The yield was 29.06%. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 13.04 (s, 1H), 9.02 (s, 1H), 7.68 (dd, J = 13.4Hz, 2.2Hz, 1H), 7.38 (t, J = 8.8Hz, 1H), 7.11 (d, J = 8.8Hz, 1H), 6.86 (d, J = 8.0 Hz,2H),6.80(d,J=8.0Hz,2H),6.55(d,J=7.6Hz,1H),4.49(s,2H),3.51-3. 49(m,2H),1.99(d,J=9.6Hz,2H),1.89(d,J=9.6Hz,2H),1.46-1.23(m,4H). 13C NMR(101MHz,DMSO-d6)δ171.46,155.36,154.74,152.91,152.60,151.84,142.12,142.02,128.75,128.62,124.65,124 .52,121.97,119.42,117.47,115.78,113.93,106.06,105.83,75.32,66.10,47.72,30.34,30.26.ESIMS:m / z484.8[MH] - .

[0122] Example 17 Synthesis of ethyl acetate 2-(4-{(1r,4r)-4-[(tert-butoxycarbonyl)amino]cyclohexyl}oxy)phenoxy)ethyl acetate

[0123] To a 25 mL single-necked flask, methyl 2-(4-{(1r,4r)-4-[(tert-butyloxycarbonyl)amino]cyclohexyl)oxy}phenoxy)acetate (4.00 g, 13.02 mmol, 1 eq), MeCN (20 mL), K₂CO₃ (5.40 g, 39.06 mmol, 3 eq), and KI (0.22 g, 1.30 mmol, 0.1 eq) were added dropwise. Ethyl chloroacetate (2.39 g, 19.53 mmol, 1.5 eq) was added dropwise, followed by Ar displacement three times, and the mixture was heated to reflux. After 16 hours, the reaction was stopped by TLC monitoring, concentrated to dryness under reduced pressure, and extracted with water (30 mL), DCM (30 mL × 3). The organic layers were combined, washed with water (30 mL), washed with saturated brine (30 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain 0.78 g of a brownish-black oily liquid. The crude product was purified by silica gel column chromatography. The column was packed with 1.5 times the volume of sample and 3 times the volume of silica gel. The eluent was EA:PE = 1:5, yielding 4.06 g of a white solid.

[0124] Example 18 Synthesis of ethyl acetate 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)

[0125] To a 100 mL single-necked flask, ethyl acetate (4.06 g, 10.33 mmol) and DCM (20 mL) were added sequentially. The mixture was cooled to 0 °C in an ice bath, and TFA (6 mL) was added dropwise. After 6 hours, TLC monitoring showed that the reaction was complete, and the reaction was stopped. The DCM and TFA in the reaction solution were removed by vacuum distillation, yielding 4.40 g of crude brown oil. The crude product was used directly in the next step without purification.

[0126] Example 19 Synthesis of 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)ethyl acetate (SP-C04)

[0127] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a 15 mL solution of 3-fluoro-4-trifluoromethoxyaniline (0.21 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) in DCM dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0128] A 25 mL solution of DCM in 3-fluoro-4-trifluoromethoxyaniline isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) were added to a 100 mL three-necked flask. The mixture was kept on ice at 0 °C, and a 10 mL solution of DCM in ethyl acetate (0.23 g, 0.79 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed the reaction was complete after 30 min, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 0.62 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 12x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:10, yielding 0.27 g of white solid (69.23% yield). 1 H NMR(400MHz, DMSO-d6)δ8.73(s,1H),7.68(dd,J=13.4,2.5Hz,1H),7.44-7.34(m,1H),7.09(ddd,J=9.0,2.6,1.3Hz,1H),6.92-6.79(m,4H),6.31(d, J=7.6Hz,1H),4.69(s,2H),4.16(q,J=7.1Hz,3H),3.55-3.45(m,1H),2.05 -1.97(m,2H),1.95-1.87(m,2H),1.50-1.25(m,4H),1.21(t,J=7.1Hz,3H). 13C NMR(101MHz,DMSO-d6)δ169.44,154.63,152.93,152.18,141.95,141.84,128.85,124.73,121.97,1 17.48,115.98,114.01,106.15,105.92,75.29,65.72,61.00,47.78,30.38,30.27,14.52.ESIMS:m / z 537.1[M+Na] + .

[0129] Example 20 Synthesis of methyl 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)-2-methylpropionate (SP-C06)

[0130] A 10 mL solution of DCM in 3-fluoro-4-trifluoromethoxyaniline isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) were added to a 25 mL three-necked flask. The mixture was kept on ice at 0 °C, and a 10 mL solution of DCM in 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)-2-methylpropionate (0.24 g, 0.79 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed the reaction was complete after 30 min, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 0.47 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 10x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:5, yielding 0.32 g of white solid (76.19% yield). 1 HNMR(400MHz,DMSO-d6)δ8.73(s,1H),7.68(dd,J=13.5,2.5Hz,1H),7.39(t,J =8.9Hz,1H),7.09(dt,J=9.3,1.9Hz,1H),6.89-6.81(m,2H),6.79-6.71(m,2H ),6.30(d,J=7.6Hz,1H),4.21(dt,J=9.9,5.6Hz,1H),3.70(s,3H),3.50(s,1H ),3.43-3.36(m,2H),2.01(d,J=15.5Hz,2H),1.95-1.87(m,2H),1.45(s,10H). 13C NMR(101MHz,DMSO-d6)δ174.26,154.63,153.28,148.83,141.94,141.84,128.85,124.72,121.97,121.76,1 19.42,116.85,114.01,106.16,105.92,79.60,75.05,52.69,47.79,30.42,30.31,25.37,15.62.ESIMS:m / z 551.2[M+Na] + .

[0131] Example 21 Synthesis of 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)-2-methylpropionic acid (SP-C05)

[0132] Methyl 2-[4-((1r,4r)-4-{3-[3-fluoro-4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)-2-methylpropionate (0.20 g, 0.38 mmol, 1 eq), THF (5 mL), H2O (0.5 mL), and LiOH (30 mg, 1.20 mmol, 3 eq) were added sequentially to a 25 mL single-necked flask and stirred at room temperature. After 3 h, the reaction was stopped by TLC monitoring. The THF was removed by concentration under reduced pressure, and DCM (30 mL) was added. The mixture was extracted with water (10 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to give 0.15 g of white solid. The yield was 76.92%. 1 H NMR (400MHz, DMSO-d6) δ12.93(s,1H),8.78(s,1H),7.68(dd,J=13.5,2.5Hz,1H ),7.44-7.35(m,1H),7.09(ddd,J=9.1,2.6,1.3Hz,1H),6.90-6.82(m,2H),6.82 -6.75(m,2H),6.34(d,J=7.6Hz,1H),4.21(dq,J=9.6,5.6,4.7Hz,1H),3.57-3. 46(m,1H),2.05-1.97(m,2H),1.91(dd,J=12.8,4.2Hz,2H),1.50-1.26(m,10H). 13C NMR(101MHz,DMSO-d6)δ175.60,155.37,154.64,152.93,149.26,141.96,141.86,124.71,121.9 7,121.32,116.88,114.00,106.14,105.90,79.27,75.08,47.77,30.40,30.29,25.46.ESIMS:m / z 512.8[MH] - .

[0133] Example 22 Synthesis of methyl 2-[4-((1r,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)acetate (SP-C09)

[0134] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a 20 mL solution of p-trifluoromethoxyaniline (0.14 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) in DCM dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0135] A 10 mL solution of DCM in p-trifluoromethoxyaniline isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) were added to a 25 mL three-necked flask. The mixture was kept on ice at 0 °C, and a 10 mL solution of DCM in 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)methyl acetate (0.22 g, 0.79 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring was performed after 30 min, and the reaction was stopped when complete. The DCM was removed by concentration under reduced pressure, yielding 0.50 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 10x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:10, yielding 0.31 g of white solid.

[0136] 1H NMR (400MHz, DMSO-d6) δ8.51(s,1H),7.51-7.43(m,2H),7.21(d,J=8.2Hz,2H),6.91-6.80(m,4H),6.18(d,J=7.6Hz,1H),4.71(s,2H ),4.19(dt,J=9.8,5.7Hz,1H),3.69(s,3H),3.57-3.46(m,1H),2.00(dd,J=11.5,4.1Hz,2H),1.96-1.87(m,2H),1.49-1.26(m,4H). 13 C NMR(101MHz,DMSO-d6)δ169.93,154.87,152.21,152.17,142.43,140.29,122.0 9,119.01,117.50,115.95,75.34,65.62,52.19,47.71,30.48,30.29.ESIMS:m / z 505.0[M+Na] + .

[0137] Example 23 Synthesis of 2-[4-((1r,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)acetic acid (SP-C08)

[0138] To a 25 mL single-necked flask, methyl 2-[4-((1r,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)acetate (0.20 g, 0.41 mmol, 1 eq), THF (5 mL), H₂O (0.5 mL), and LiOH (30 mg, 1.20 mmol, 3 eq) were added sequentially and stirred at room temperature. After 3 h, the reaction was stopped by TLC monitoring. The mixture was concentrated under reduced pressure to remove THF, and extracted with DCM (30 mL) and water (10 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to obtain 0.15 g of a white solid. The yield was 78.94%. 1 H NMR (400MHz, DMSO-d6) δ8.76(s,1H),7.51-7.44(m,2H),7.21(d,J=8.6Hz,2H),6.91-6.76(m,4H),6.40(d,J=7.6Hz,1H),4.52(s,2H) ,4.17(tt,J=9.7,4.0Hz,1H),3.55-3.45(m,1H),2.04-1.95(m,2H),1.94-1.86(m,2H),1.36(dddd,J=40.3,20.2,16.5,10.0Hz,5H). 13C NMR(101MHz,DMSO-d6)δ171.14,154.96,152.56,151.89,142.35,140.44,12 2.04,118.97,117.49,115.79,75.39,66.00,47.67,30.45,30.29.ESIMS:m / z 466.8[MH] - .

[0139] Example 24 Synthesis of 2-[4-((1r,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)ethyl acetate (SP-C10)

[0140] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a 15 mL solution of p-trifluoromethoxyaniline (0.14 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) in DCM dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. After 10 minutes, monitor the reaction by TLC (EA:PE = 1:3), indicating that the reaction is complete. Stop the reaction.

[0141] A 10 mL solution of DCM in p-trifluoromethoxyaniline isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) were added to a 25 mL three-necked flask. The flask was kept on ice at 0 °C, and a 10 mL solution of DCM in ethyl acetate 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)acetate (0.23 g, 0.79 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring was performed after 30 min, and the reaction was stopped when complete. The DCM was removed by concentration under reduced pressure, yielding 0.93 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 12x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:10, yielding 0.24 g of white solid.

[0142] 1 H NMR (400MHz, DMSO-d6) δ8.52(s,1H),7.50-7.43(m,2H),7.22(d,J=8.6Hz,2H),6.92-6.80(m,4H),6.18(d,J=7.6Hz,1H),4.69(s ,2H),4.19-4.09(m,2H),3.60-3.42(m,1H),2.04-1.96(m,2H),1.92(d,J=13.7Hz,2H),1.48-1.24(m,5H),1.21(t,J=7.1Hz,4H). 13C NMR(101MHz,DMSO-d6)δ169.43,154.87,152.20,142.43,140.28,122.08,119.0 2,117.49,115.98,75.34,65.74,61.00,47.70,30.48,30.29,14.51.ESIMS:m / z 519.1[M+Na] + .

[0143] Example 25 Synthesis of methyl 2-methyl-2-[4-((1,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)-2-methylpropionate (SP-C12)

[0144] A 10 mL solution of DCM in p-trifluoromethoxyaniline isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) were added to a 25 mL three-necked flask. The mixture was kept on ice at 0 °C, and a 10 mL solution of DCM in methyl 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)-2-methylpropionate (0.24 g, 0.79 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed the reaction was complete after 30 min, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 0.47 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 10x silica gel using 1.5x stirred sample and eluent of EA:PE = 1:5, yielding 0.33 g of white solid (76.19% yield). 1 H NMR (400MHz, DMSO-d6) δ8.51 (s, 1H), 7.50-7.42 (m, 2H), 7.21 (d, J = 8.5Hz, 2H),6.88-6.81(m,2H),6.79-6.72(m,2H),6.18(d,J=7.6Hz,1H),4.21(tt ,J=9.7,3.7Hz,1H),3.70(s,3H),3.51(dtd,J=10.9,7.3,4.2Hz,1H),2.01 (dd,J=12.6,4.6Hz,2H),1.92(dd,J=13.0,4.3Hz,2H),1.51-1.25(m,11H). 13 C NMR(101MHz,DMSO-d6)δ174.25,154.88,153.32,148.84,142.45,140.29,122.07,1 21.78,119.03,116.88,79.62,75.12,52.68,47.72,30.52,30.33,25.38.ESIMS:m / z 533.0[M+Na] + .

[0145] Example 26 Synthesis of 2-methyl-2-[4-((1,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureoyl}cyclohexyl)oxy]phenoxy)-2-methylpropionic acid (SP-C11)

[0146] To a 25 mL single-necked flask, 2-methyl-2-[4-((1,4r)-4-{3-[4-(trifluoromethoxy)phenyl]ureido}cyclohexyl)oxy]phenoxy)-2-methylpropionic acid (0.20 g, 0.39 mmol, 1 eq), THF (5 mL), H2O (0.5 mL), and LiOH (30 mg, 1.20 mmol, 3 eq) were added sequentially and stirred at room temperature. After 3 h, the reaction was stopped by TLC monitoring. The mixture was concentrated under reduced pressure to remove THF, and extracted with DCM (30 mL) and water (10 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to obtain 0.14 g of a white solid. The yield was 73.68%.

[0147] 1 H NMR (400MHz, DMSO-d6) δ8.54(s,1H),7.51-7.43(m,2H),7.22(d,J=8.6Hz,2H),6.90-6.72(m,4H),6.20(d,J=7.6Hz,1H ),4.21(td,J=9.6,4.8Hz,1H),3.57-3.45(m,1H),2.08-1.97(m,2H),1.91(dd,J=12.0,4.6Hz,2H),1.50-1.23(m,11H). 13 C NMR(101MHz,DMSO-d6)δ175.60,154.88,152.94,149.26,142.42,140.30,122.0 8,121.31,119.01,116.88,79.27,75.13,47.70,30.51,30.32,25.46.ESIMS:m / z 494.9[MH] - .

[0148] Example 27 Synthesis of methyl acetate 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}acetate (SP-C15)

[0149] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a 20 mL solution of a mixture of ammonium phosphate (0.14 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) in DCM dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0150] Add 10 mL of DCM solution of p-methammonium isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) to a 25 mL three-necked flask. Heat to 0 °C on ice and add 10 mL of DCM solution of methyl 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)acetate (0.22 g, 0.79 mmol, 1 eq). After addition, allow the reaction to proceed at room temperature. After 30 min, monitor the reaction by TLC. Once the reaction is complete, stop the reaction. Concentrate under reduced pressure to remove DCM, yielding 0.50 g of crude white solid. Purify the crude product by silica gel column chromatography. Mix 1.5 times the sample and pack a 10x silica gel column with EA:PE = 1:10 as eluent to obtain 0.33 g of white solid.

[0151] 1 H NMR (400MHz, DMSO-d6) δ6.89-6.79 (m, 4H), 5.58 (d, J = 7.6Hz, 1H), 5.41 (s, 1H), 4.70(s,2H),4.15(tt,J=10.0,4.0Hz,1H),3.69(s,3H),3.35(dt,J=7.3,3.6Hz ,1H),2.04(p,J=3.2Hz,1H),1.96(dt,J=13.4,3.9Hz,2H),1.86-1.77(m,2H),1 .67(d,J=3.1Hz,2H),1.49(s,4H),1.40-1.11(m,9H),1.07(s,2H),0.79(s,6H). 13 C NMR (101MHz, DMSO-d6) δ169.92,157.00,152.25,152.17,117.56,115.96,75.50,65.66,52. 18,51.49,50.87,48.61,47.37,42.91,41.00,32.36,30.83,30.61,30.40,30.10.ESIMS:m / z 507.1[M+Na] + .

[0152] Example 28 Synthesis of 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}acetic acid (SP-C14)

[0153] To a 25 mL single-necked flask, methyl 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}acetate (0.20 g, 0.41 mmol, 1 eq), THF (5 mL), H2O (0.5 mL), and LiOH (30 mg, 1.24 mmol, 3 eq) were added sequentially and stirred at room temperature. After 3 h, the reaction was stopped by TLC monitoring. The THF was removed by concentration under reduced pressure, and DCM (30 mL) was added. The mixture was extracted with water (10 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to obtain 0.15 g of white solid.

[0154] 1 H NMR (400MHz, DMSO-d6) δ6.85-6.78(m,2H),6.78-6.70(m,2H),5.77(d,J=7.6Hz,1H),5.5 9(s,1H),4.24(s,2H),4.11(tt,J=9.5,4.0Hz,1H),3.33(ddd,J=11.1,7.1,3.7Hz,1H),2 .04(p,J=3.2Hz,1H),2.00-1.90(m,2H),1.81(dq,J=11.8,3.8Hz,2H),1.67(d,J=3.1Hz, 2H),1.55-1.43(m,4H),1.42-1.25(m,4H),1.25-1.09(m,5H),1.07(s,2H),0.79(s,6H). 13 C NMR(101MHz,DMSO-d6)δ171.28,157.12,153.18,151.45,117.53,115.68,75.61,67 .74,51.46,50.89,48.61,47.33,42.93,40.99,32.36,30.79,30.62,30.40,30.10.

[0155] Example 29 Synthesis of 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}ethyl acetate (SP-C16)

[0156] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a DCM solution of ammonium phosphate (0.14 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0157] Add 10 mL of DCM solution of memantine isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) to a 25 mL three-necked flask. Heat to 0 °C on ice and add 10 mL of DCM solution of ethyl acetate 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)acetate (0.23 g, 0.79 mmol, 1 eq). After addition, react at room temperature. After 30 min, monitor the reaction by TLC. Once the reaction is complete, stop the reaction. Concentrate under reduced pressure to remove DCM, yielding 0.93 g of crude white solid. Purify the crude product by silica gel column chromatography. Mix 1.5 times the sample and pack a 15 times silica gel column (to remove memantine that does not fluoresce and is converted to urea by itself). The eluent is EA:PE = 1:10, yielding 0.20 g of white solid.

[0158] 1 H NMR(400MHz, DMSO-d6)δ6.89-6.79(m,4H),5.58(d,J=7.6Hz,1H),5.41(s,1H),4.68(s,2H),4.16(q,J=7.1Hz,3H),3.35(dd,J=7.1,3.6Hz,1H), 2.04(p,J=3.2Hz,1H),2.01-1.90(m,2H),1.86-1.76(m,2H),1.67(d,J= 3.2Hz,2H),1.49(s,4H),1.40-1.11(m,11H),1.07(s,2H),0.79(s,6H). 13 C NMR(101MHz,DMSO-d6)δ169.43,156.99,152.21,152.17,117.51,115.97,75.46,65.73,61 .00,51.47,50.85,48.60,47.36,42.90,40.99,32.36,30.84,30.61,30.40,30.09,14.52.

[0159] Example 30 Synthesis of methyl 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}-2-methylpropionate (SP-C18)

[0160] Add BTC (0.08 g, 0.27 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Then, add a DCM solution of ammonium phosphate (0.14 g, 0.79 mmol, 1 eq) and Et3N (0.24 g, 2.37 mmol, 3 eq) dropwise over 30 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. Monitor the reaction by TLC after 10 minutes. Once the reaction is complete, stop the reaction.

[0161] Add 10 mL of DCM solution of memantine isocyanate and Et3N (0.24 g, 2.37 mmol, 3 eq) to a 25 mL three-necked flask. Heat to 0 °C on ice and add 10 mL of DCM solution of methyl 2-{4-[(1r,4r)-4-aminocyclohexyl]oxy}phenoxy)-2-methylpropionate (0.24 g, 0.79 mmol, 1 eq). After addition, react at room temperature. After 30 min, monitor the reaction by TLC. Once the reaction is complete, stop the reaction. Concentrate under reduced pressure to remove DCM, yielding 0.93 g of crude white solid. Purify the crude product by silica gel column chromatography. Mix 1.5 times the sample and pack a 15 times silica gel column with EA:PE = 1:10 as eluent to obtain 0.32 g of white solid.

[0162] 1 H NMR (400MHz, DMSO-d6) δ6.87-6.78(m,2H),6.78-6.69(m,2H),5.58(d,J=7.6Hz,1H),5.41(s,1H),4.22-4.11(m,1H),3.69(s,3H), 2.04(p,J=3.1Hz,1H),2.01-1.92(m,2H),1.86-1.77(m,2H),1.67(d,J=3.2Hz,2H),1.50-1.13(m,18H),1.07(s,2H),0.79(s,6H). 13 C NMR(101MHz,DMSO-d6)δ174.25,156.99,153.32,148.80,121.77,116.88,79.61,75.21,52 .68,51.48,50.86,48.60,47.37,42.91,40.99,32.36,30.88,30.61,30.44,30.09,25.38.

[0163] Example 31 Synthesis of 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}-2-methylpropionic acid (SP-C17)

[0164] Methyl 2-[4-((1R,4r)-4-{3-[(1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl]ureido}cyclohexyl)oxy]phenoxy}-2-methylpropionate (0.20 g, 0.39 mmol, 1 eq), THF (5 mL), H2O (0.5 mL), and LiOH (30 mg, 1.24 mmol, 3 eq) were added sequentially to a 25 mL single-necked flask and stirred at room temperature. After 3 h, the reaction was stopped by TLC monitoring. The THF was removed by concentration under reduced pressure, and DCM (30 mL) was added. The mixture was extracted with water (10 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl and filtered to obtain 0.10 g of white solid.

[0165] 1 H NMR (400MHz, DMSO-d6) δ6.79(s,4H),5.73(d,J=7.7Hz,1H),5.54(s,1H),4.14(td,J=9.6,4.8Hz,1H),3.33(ddd,J=10.8,7.0,3.6Hz,1H),2.04( p,J=3.2Hz,1H),2.01-1.91(m,2H),1.85-1.76(m,2H),1.67(d,J=3.2Hz ,2H),1.55-1.43(m,4H),1.43-1.10(m,14H),1.07(s,2H),0.79(s,6H). 13 C NMR(101MHz,DMSO-d6)δ175.96,157.08,152.49,149.72,121.01,116.83,79.75,75.30 ,51.46,50.88,48.60,47.34,42.92,40.98,32.36,30.84,30.62,30.43,30.09,25.78.

[0166] Example 32 Synthesis of 4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl acetate

[0167] Add 1.69 g (4.84 mmol) of 4-(((1r, 4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)phenyl acetate and 12 mL of DCM to a 100 mL single-necked flask, and then add 6 mL of TFA dropwise at room temperature. After 3 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. Remove TFA from the reaction solution by vacuum distillation. 2.06 g of crude brown oil was obtained. The crude product was dried in a 60 °C oven for 12 h and used directly in the next step without purification.

[0168] Example 33 Synthesis of 4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenyl acetate (SP-C01)

[0169] Add BTC (0.51 g, 1.17 mmol, 0.34 eq) and dry DCM (10 mL) sequentially to a 100 mL single-necked flask. Cool the flask to -78 °C in a cold trap, then add 3-fluoro-4-trifluoromethoxyaniline (1 g, 5.13 mmol, 1 eq), Et3N (1.65 g, 15.39 mmol, 3 eq), and a DCM solution (10 mL) dropwise over 30 minutes. After the addition is complete, bring the flask to room temperature and continue stirring for 4 hours. After 4 hours, monitor the reaction by TLC. Take one drop of the reaction solution into an EP tube, add memantine (EA:PE = 1:3), fumigate with iodine, and allow the reaction to complete.

[0170] A mixed solution of 4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl acetate·TFA (1.86 g, 5.13 mmol, 1 eq), Et3N (1.65 g, 15.39 mmol, 3 eq), and DCM (5 mL) was added to a 100 mL single-necked flask. The above reaction solution was added dropwise, and the reaction was carried out at room temperature. After 2 hours, the reaction was monitored by TLC (EA:PE = 1:3, AcOH 2d), indicating that the reaction was complete. The reaction was stopped. The sample was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.23 g of crude yellow oil. The sample was packed into a 5x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:10) to give 0.42 g of white solid, with a yield of 17.3%. 1H NMR (400MHz, DMSO-d6): δ (ppm) 8.727.69 (d, J = 2.5Hz, 1H), 7.65-7.38 (m, 1H), 7.36-7.35 (m, 1H), 7.11-6.94 (m, 4H), 6.30 (d, J=8.0Hz,1H),4.29(d,J=4.0Hz,1H),3.51(s,1H),2.23(s,3H),2.05(d,J=3.0Hz,2H),2.02-1.91(m,2H),1.48-1.34(m,4H). 13 C NMR (100MHz, DMSO-d6): δ (ppm) 169.9, 155.3, 154.6, 152.9, 144.3, 141.9, 124.7, 1 23.1,116.8,114.0,114.0,106.2,105.9,75.0,47.8,30.4,30,2,21.1.ESIMS:m / z 471.1[M+H] + .

[0171] Example 34 Synthesis of 4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenol (SP-C01b)

[0172] To a 100 mL single-necked flask, 4-(((1r, 4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureido)cyclohexyl)oxy)phenyl acetate (0.24 g, 0.51 mmol, 1 eq), LiOH (2.1 g, 17.20 mmol, 34 eq), and H₂O (7 mL) were added sequentially, and the reaction was carried out at room temperature. After 1.5 hours, TLC monitoring showed that the reaction was complete, and the reaction was stopped. The mixture was extracted with DCM (30 mL × 3), and the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. 0.21 g of crude brown solid was given. Yield: 96.33%.

[0173] Example 35 Synthesis of methyl 4-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)benzoate

[0174] Add cis-4-BOC-aminocyclohexanol (1.00 g, 4.64 mmol), methylparaben (0.7 g, 4.64 mmol), triphenylphosphine (1.8 g, 6.96 mmol), A4 molecular sieve (dried in an oven at 120 °C for 4 h), and THF (10 mL) sequentially to a 100 mL single-necked flask. Purge the solution three times with argon gas, then cool to below -10 °C in an ice-salt bath. Add 5 mL of a THF solution containing DIAD (1.4 g, 6.96 mmol) dropwise at a rate of one drop every two seconds. After 12 hours, TLC monitoring showed an EA:PE ratio of 1:1. Move the reaction vessel to 30 °C and add more 4A molecular sieve. After 6 hours, TLC monitoring showed an EA:PE ratio of 1:1, indicating the reaction was largely complete. Stop the reaction. THF was removed by vacuum concentration, yielding 5.2 g of brownish-yellow oily substance. No solid precipitate was observed after pulping. The sample was packed into a column with 4x silica gel, mixed with 1.2x silica gel, and eluent (EA:PE = 1:10), yielding 0.73 g of white solid product, with a yield of 45.1%.

[0175] Example 36 Synthesis of methyl 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzoate

[0176] Methyl benzoate (0.60 g, 2.09 mmol) and DCM (5 mL) were added sequentially to a 100 mL single-necked flask, followed by dropwise addition of TFA (4 mL) at room temperature. After 3 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. TFA was removed from the reaction solution by vacuum distillation. 0.62 g of crude brown oil was obtained. The crude product was dried in a 60 °C oven for 12 h and used directly in the next step without purification.

[0177] Example 37 Synthesis of methyl 4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoate

[0178] Add BTC (0.23 g, 0.79 mmol) and dry DCM (15 mL) sequentially to a 100 mL single-necked flask. Cool the flask to -78 °C and add free 3-fluoro-4-trifluoromethoxyaniline (0.41 g, 2.32 mmol), Et3N (1.41 g, 13.92 mmol), and a DCM solution (10 mL) dropwise over 30 minutes. After the addition is complete, bring the flask to room temperature and continue stirring for 4 h. After 4 h, monitor the reaction by TLC. Take one drop of the reaction solution into an EP tube and add methyl 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzoate, EA:PE = 1:3.

[0179] A mixed solution of methyl 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzoate·TFA (0.58 g, 1.59 mmol), Et3N (1.41 g, 13.92 mmol), and DCM (10 mL) was added to a 100 mL single-necked flask. The reaction was carried out at room temperature, and after 2 hours, the reaction was stopped by TLC monitoring (EA:PE = 1:3). The mixture was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.93 g of crude yellow oil. The solution was packed into a 7x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:5) to give 0.40 g of the product as a white solid, yield 40.0%. ESI-MS: m / z 453.2 [M+H] +

[0180] Example 38 Synthesis of 4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoic acid (SP-C01c)

[0181] Methyl 4-(((1r, 4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoate (2.59 g, 5.50 mmol, 1 eq), THF (20 mL), H2O (2 mL), and LiOH (0.40 g, 16.50 mmol, 3 eq) were added sequentially to a 100 mL single-necked flask and stirred at room temperature. After 3 h, TLC monitoring showed that (EA:PE = 1:1), the reaction was stopped, and the mixture was concentrated under reduced pressure to remove THF. DCM (50 mL) was added, and the mixture was extracted with water (15 mL × 2). The aqueous layer was adjusted to pH 2 with 1 N HCl, concentrated under reduced pressure until a small amount of solvent remained, filtered, and washed with 5 mL of water to give 0.53 g of a pale yellow solid. The combined yield of the three steps was 21.13%.

[0182] 1 H NMR (600MHz, DMSO-d6) δ10.19(s,1H),8.88(s,1H),8.35(s,3H),7.86(d,J=8.6Hz,2H),7.78(d,J=8.5Hz,1H),7.04(d,J=8.6Hz,5H),6.85(d,J=8. 3Hz,4H),4.39(tt,J=10.0,4.3Hz,1H),3.04(dt,J=10.8,5.5Hz,1H),2.1 5-2.09(m,2H),2.06-2.01(m,2H),1.61-1.50(m,2H),1.50-1.38(m,2H).

[0183] Example 39 Synthesis of 4-(((1R,4r)-4-(3-(((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureo)cyclohexyl)oxy)phenyl acetate (SP-C01d)

[0184] Add BTC (0.02 g, 0.07 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 100 mL single-necked flask. Cool the flask to -78 °C in a cold trap, then add memantine (0.04 g, 0.22 mmol, 1 eq), Et3N (0.07 g, 0.66 mmol, 3 eq), and 10 mL of DCM solution dropwise over 30 minutes. After the addition is complete, bring the flask to room temperature and continue stirring for 4 hours. Monitor the reaction by TLC after 4 hours. (Take one drop of the reaction solution into an EP tube, add 4-trifluoromethoxyaniline, EA:PE = 1:3), fumigate with iodine, and the reaction is complete.

[0185] A mixture of 4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl acetate·TFA (0.08 g, 0.22 mmol, 1 eq), Et3N (0.07 g, 0.66 mmol, 3 eq), and DCM (5 mL) was added dropwise to a 25 mL single-necked flask. The reaction mixture was then added dropwise, and the mixture was reacted at room temperature. After 2 hours, the reaction was monitored by TLC. Once the reaction was complete, the reaction was stopped. The mixture was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.10 g of crude yellow oil. The solution was packed into a 5x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:10) to give 32 mg of white solid, with a yield of 32.0%. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 7.04-6.97 (d, 2H), 6.97-6.89 (d, 2H), 5.58 (d, J = 7.9Hz, 1H), 5.41 ( d,J=3.2Hz,1H),4.25(tt,J=9.9,3.9Hz,1H),3.35(dd,J=7.3,3.7Hz,1H),2.23(s,3H),2.02(ddt,J =24.4,8.4,3.5Hz,3H),1.87-1.80(m,2H),1.67(d,J=3.1Hz,2H),1.50(s,4H),1.37(ddd,J=15.2,9 .1,4.3Hz,2H),1.31-1.22(m,4H),1.22-1.16(m,2H),1.16-1.10(m,1H),1.07(s,2H),0.80(s,6H). 13C NMR (100MHz, DMSO-d6): δ (ppm) 169.94, 157.00, 155.37, 144.29, 123.06, 116.77, 75.17, 51.49, 50.87,48.61,47.33,42.91,41.00,32.36,30.89,30.82,30.61,30.31,30.10,21.26.ESIMS:m / z 455.2[M+H] + .

[0186] Example 40 Synthesis of tert-butyl ((1r,4r)-4-(4-acetylphenoxy)cyclohexyl)carbamate

[0187] Add cis-4-BOC-aminocyclohexanol (1 g, 4.64 mmol, 1 eq), acetaminophen (0.70 g, 4.64 mmol, 1 eq), PPh3 (1.83 g, 6.96 mmol, 1.5 eq), type 4A molecular sieve (dried in an oven at 120 °C for 4 h), and THF (20 mL) sequentially to a 250 mL single-necked flask. Purge the solution three times with argon gas, cool to -10 °C in an ice-salt bath, and add 10 mL of a THF solution containing DIAD (1.21 g, 6.96 mmol, 1.5 eq) dropwise at a rate of one drop every two seconds. After 12 hours, TLC monitoring showed that EA:PE = 1:2, indicating the reaction was largely complete, and the reaction was stopped. The solution was concentrated under reduced pressure to remove THF, yielding 5.62 g of a brownish-yellow oily substance. Add 10 mL of ethanol and 5 mL of petroleum ether, stir for 2 h, slurry, and filter to obtain 0.90 g of a pale yellow solid, with a yield of 55.9%.

[0188] Example 41 Synthesis of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)acetamide

[0189] To a 100 mL single-necked flask, tert-butyl ((1r,4r)-4-(4-acetylphenoxy)cyclohexyl)carbamate (0.90 g, 2.58 mmol) and DCM (6 mL) were added sequentially, followed by dropwise addition of TFA (4 mL) at room temperature. After 3 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. TFA was removed from the reaction solution by vacuum distillation. 1.06 g of crude brown oil was obtained. This was dried in a 60 °C oven for 12 h. The crude product was used directly in the next step without purification.

[0190] Example 42 Synthesis of N-(4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenyl)acetamide (SP-C01e)

[0191] Add BTC (0.08 g, 0.26 mmol, 0.34 eq) and dry DCM (10 mL) sequentially to a 50 mL single-necked flask. Cool the flask to -78 °C and add dropwise a DCM solution of 3-fluoro-4-trifluoromethoxyaniline (0.15 g, 0.77 mmol, 1 eq) and Et3N (0.47 g, 4.61 mmol, 3 eq) over 15 minutes. After the addition is complete, bring the flask to room temperature and continue stirring for 0.5 h. After 0.5 h, monitor the reaction by TLC. Take one drop of the reaction solution into an EP tube and add N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)acetamide·TFA, EA:PE = 1:1.

[0192] A mixed solution of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)acetamide·TFA (0.30 g, 0.77 mmol), Et3N (0.47 g, 4.61 mmol), and DCM (5 mL) was added to a 100 mL single-necked flask. The reaction was carried out at room temperature, and the reaction was stopped after 0.5 hours by TLC monitoring (EA:PE = 1:1). The sample was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.52 g of crude yellow oil. The sample was packed into a 4x silica gel column, mixed with 1.2x silica gel, and eluented with (EA:PE = 1:5, EA:PE = 1:3) to give 0.11 g of white solid, yield 33.4%. ESI-MS: m / z 470.2 [M+H] + 492.1 [M+Na] +

[0193] 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 9.75 (s, 1H), 8.74 (s, 1H), 7.67 (dd, J = 13.4, 2.4H z,1H),7.44(d,J=8.9Hz,2H),7.38(t,J=8.7Hz,1H),7.09(d,J=8.8Hz,1H),6.86(d ,J=8.9Hz,2H),6.29(d,J=7.6Hz,1H),4.26-4.21(m,1H),3.53-3.48(m,1H),2.03- 2.20(m,2H),2.20(s,3H),1.93-1.90(m,2H),1.46-1.38(m,2H),1.38-1.32(m,2H).

[0194] Example 43 Synthesis of N-(4-hydroxyphenyl)-2-methylbutyramide

[0195] Add 2-methylbutyric acid (3.0 g, 27.51 mmol, 1 eq) and dry tetrahydrofuran (15 mL) to a 100 mL flask. Heat to 0 °C on ice, then add SOCl2 (3.93 g, 33.01 mmol, 1.2 eq) dropwise and stir for 30 min. Transfer a portion of the solution to a 1.5 mL EP tube, add anhydrous methanol, monitor the reaction for completion by TLC, and concentrate to dryness under reduced pressure.

[0196] Take a 100 mL round-bottom flask, add p-aminophenol (3.0 g, 27.51 mmol, 1 eq), THF (5 mL), and add dropwise a THF solution of 2-methylbutyryl chloride (10 mL). After the addition is complete, monitor the reaction by TLC. If the reaction is complete and the origin is the oxidation point of the starting material, stop the reaction. Concentrate under reduced pressure to remove tetrahydrofuran, extract twice with dichloromethane (20 mL) and water (20 mL), wash once with saturated brine (25 mL), dry with anhydrous magnesium sulfate, filter, and concentrate the organic phase under reduced pressure to obtain 16.5 g of yellow oil. Pack a column with 4x silica gel, add 1.2x silica gel as sample, and elute with EA:PE = 1:5. Column chromatography yields 4.85 g of white solid. The yield is 91.3%. ESI-MS: m / z 194.1 [M+H] + 216.1 [M+Na] +

[0197] Example 44 Synthesis of tert-butyl ((1r,4r)-4-(4-(2-methylbutamido)phenoxy)cyclohexyl)carbamate

[0198] Add cis-4-BOC-aminocyclohexanol (2.00 g, 9.29 mmol, 1 eq), N-(4-hydroxyphenyl)-2-methylbutyramide (1.79 g, 9.29 mmol, 1 eq), triphenylphosphine (3.65 g, 13.9 mmol, 1.5 eq), A4 molecular sieve (dried in an oven at 120 °C for 4 h), and THF (15 mL) sequentially to a 100 mL single-necked flask. Purge the solution three times with argon gas, cool to below -10 °C in an ice-salt bath, and then add DIAD (2.43 g, 13.9 mmol, 1.5 eq) in THF solution (5 mL) dropwise at a rate of one drop every two seconds. After the addition is complete, transfer the flask to room temperature. After 12 hours, TLC monitoring showed that EA:PE = 1:1, indicating that the reaction was largely complete, and the reaction was stopped. To remove some THF, the solid was concentrated under reduced pressure. 15 mL of anhydrous ethanol and 15 mL of diethyl ether were added and the mixture was stirred. The solid was filtered to obtain 2.24 g of light pink solid. The solid was monitored by TLC. 20 mL of DCM was added to dissolve the solid. The solid was washed with 16 mL of saturated Na2CO3. The organic layer was dried over anhydrous magnesium sulfate. The solid was filtered and concentrated under reduced pressure to dryness to obtain 1.94 g of pure white solid, with a yield of 53.6%.

[0199] Example 45 Synthesis of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)-2-methylbutyramide

[0200] To a 100 mL single-necked flask, tert-butyl((1r,4r)-4-(4-(2-methylbutamido)phenoxy)cyclohexyl)carbamate (1.94 g, 4.97 mmol) and DCM (5 mL) were added sequentially, followed by dropwise addition of TFA (6 mL) at room temperature. After 3 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. The reaction solution was concentrated under reduced pressure to remove TFA. 1.92 g of crude brown oily N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)-2-methylbutamidoamide·TFA was obtained. The crude product was dried in an oven at 60 °C for 12 h and used directly in the next step without purification.

[0201] Example 46 Synthesis of N-(4-(((1R,4r)-4-(3-(((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureo)cyclohexyl)oxy)phenyl)acetamide (SP-C01f)

[0202] Add BTC (0.14 g, 0.47 mmol) and dry DCM (10 mL) sequentially to a 100 mL single-necked flask. Cool the flask to -78 °C and add free memantine (0.25 g, 1.41 mmol), Et3N (0.85 g, 8.46 mmol), and DCM solution (10 mL) dropwise over 30 minutes. After the addition is complete, move the flask to room temperature and continue stirring for 4 h.

[0203] A mixed solution of A4 (0.35 g, 1.41 mmol), Et3N (0.28 g, 2.82 mmol), and DCM (5 mL) was added to a 100 mL single-necked flask and reacted at room temperature. After 2 hours, the reaction was stopped by TLC monitoring. The mixture was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.52 g of crude yellow oily product. The product was packed into a 4x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:5) to give 0.21 g of product, with a yield of 36.7%.

[0204] 1H-NMR (400MHz, DMSO-d6): δ (ppm) 9.73 (s, 1H), 7.43 (d, J = 8.96Hz, 2H), 6.84 (d, J = 8.96Hz, 2H) ,5.54(d,J=7.60Hz,1H),5.41(s,1H),4.22-4.17(m,1H),3.37-3.34(m,1H),2.05-2.03(m,1H) ,1.99(s,3H),1.97-1.94(m,1H),1.84-1.81(m,2H),1.67-1.66(m,2H),1.52-1.46(m,4H),1. 40-1.34(m,2H),1.30(s,1H),1.27-1.24(m,4H),1.19-1.14(m,2H),1.07(s,2H),0.79(s,6H).

[0205] Example 47 Synthesis of N-(4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzene)-2-methylbutyramide (SP-C01g)

[0206] Add BTC (0.10 g, 0.35 mmol, 1 eq) and dry DCM (10 mL) sequentially to a 50 mL single-necked flask. Cool the flask to -78 °C and add dropwise a DCM solution of 3-fluoro-4-trifluoromethoxyaniline (0.2 g, 1.02 mmol, 1 eq) and Et3N (0.93 g, 9.22 mmol, 3 eq) over 30 minutes. After addition, bring the flask to room temperature and continue stirring for 0.5 h. Monitor the reaction by TLC. Take one drop of the reaction solution into an EP tube and add memantine.

[0207] A mixed solution of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)-2-methylbutyramide·TFA (0.41 g, 1.02 mmol, 1 eq), Et3N (0.93 g, 9.22 mmol, 3 eq), and DCM (10 mL) was added to a 100 mL single-necked flask. The reaction was carried out at room temperature, and the reaction was stopped after 0.5 hours by TLC monitoring (EA:PE = 1:2, AcOH 1 d). Extract twice with 15 mL of water, wash once with 15 mL of saturated saline, dry with anhydrous magnesium sulfate, filter, concentrate under reduced pressure to dryness, and obtain 0.74 g of crude yellow oily product. Pack a column with 7x silica gel, mix with 1.2x silica gel, and elute with EA:PE = 1:5, EA:PE = 1:2 to obtain 0.13 g of yellow solid. The purity was insufficient, so the product was placed in a 25 mL single-necked flask, dissolved with 0.5 mL of DCM, and then precipitated with 4 mL of petroleum ether. A white solid precipitated, filtered, and 62 mg of product was obtained. The filtrate was recovered, yielding 11.9%. ESI-MS: m / z 512.3 [M+H] +

[0208] 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 9.64 (s, 1H), 8.17 (s, 1H), 7.67 (dd, J = 13.4, 2.1Hz, 1H), 7.48 (d,J=8.8Hz,2H),7.38(t,J=8.9Hz,1H),7.10-7.08(m,1H),6.87(d,J=8.8Hz,2H),6.27(d,J=7 .4Hz,1H),4.26-4.22(m,1H),3.53-3.52(m,1H),2.36-2.31(m,1H),2.03-2.00(m,2H),1.93-1 .90(m,2H),1.63-1.56(m,1H),1.48-1.30(m,5H),1.06(d,J=6.7Hz,3H),0.85(t,J=7.4Hz,3H).

[0209] Example 48 Synthesis of tert-butyl ((1r,4r)-4-(4-nitrophenoxy)cyclohexyl)carbamate

[0210] Add cis-4-BOC-aminocyclohexanol (10.00 g, 0.046 mol, 1 eq), 4-nitrophenol (6.47 g, 0.046 mol, 1 eq), PPh3 (13.95 g, 0.069 mmol, 1.5 eq), and THF (10 mL) sequentially to a 100 mL three-necked flask. Cool to below -10 °C in an ice-salt bath, and add 20 mL of a THF solution containing DIAD (18.10 g, 0.069 mol, 1.5 eq) dropwise at a rate of one drop every two seconds. After 8 hours, monitor with TLC; EA:PE = 1:2, indicating the reaction is complete, and stop the reaction. Concentrate under reduced pressure to remove THF, yielding 12.21 g of a brownish-yellow oily substance. Add 50 mL of ethanol, stir for 2 hours, slurry, and filter to obtain 12.46 g of a yellow solid, yielding 80.58%.

[0211] Example 49 Synthesis of (1r,4r)-4-(4-nitrophenoxy)cyclohexane-1-amine

[0212] To a 100 mL single-necked flask, tert-butyl carbamate (0.60 g, 2.09 mmol) and DCM (5 mL) were added sequentially, followed by dropwise addition of TFA (4 mL) at room temperature. After 3 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. TFA was removed from the reaction solution by vacuum distillation. 0.62 g of crude brown oil was obtained. This crude product was dried in a 60 °C oven for 12 h and used directly in the next step without purification.

[0213] Example 50 Synthesis of 1-((1r,3R,5S,7R)-3,5-dimethyladamantane-1-yl)-3-((1r,4R)-4-(4-nitrophenoxy)cyclohexyl)urea

[0214] Add BTC (5.50 g, 18.53 mmol, 0.5 eq) and dry DCM (50 mL) sequentially to a 250 mL single-necked flask. Cool the flask to -80 °C in a cold trap. Add a 50 mL solution of DCM containing a mixture of memantine (6.56 g, 37.06 mmol, 1 eq) and Et3N (30.00 g, 296.48 mmol, 8 eq) dropwise over 4 hours. After the addition is complete, allow the flask to warm to room temperature naturally. After 30 minutes, monitor the reaction by TLC. If the reaction is complete, stop the reaction. Evaporate the reaction mixture to dryness. Add DCM (50 mL) and Et3N (30.00 g, 296.48 mmol, 8 eq). Cool the flask to 0 °C in an ice bath. Add a 50 mL solution of DCM containing I9 (8.89 g, 37.06 mmol, 1 eq) dropwise. After the addition is complete, allow the flask to warm to room temperature. After 30 minutes, monitor the reaction by TLC. If the reaction is complete, stop the reaction. DCM was removed by vacuum concentration, yielding 17.82 g of reddish-brown oily substance. The crude product was used directly in the next step without purification.

[0215] Example 51 Synthesis of 1-((1r,4R)-4-(4-aminophenoxy)cyclohexyl)-3-((1r,3R,5S,7R)-3,5-dimethyladamantane-1-yl)urea (SP-C01h)

[0216] Add 2-methylbutyric acid (3.0 g, 27.51 mmol, 1 eq), dry tetrahydrofuran (10 mL), HATU (0.034 g, 0.29 mmol, 1.2 eq), and DIEA (0.18 g, 1.74 mmol, 6 eq) to a 25 mL flask, and stir for 30 min. Add a THF solution of 1-((1r,4R)-4-(4-aminophenoxy)cyclohexyl)-3-((1r,3R,5S,7R)-3,5-dimethyladamantane-1-yl)urea (0.1 g, 0.24 mmol, 1 eq) in 5 mL. Monitor the reaction by TLC. Once the reaction is complete, stop the reaction and filter to give 72 mg of a white solid. The yield is 60.56%.

[0217] 1 H NMR(400MHz,Chloroform-d)δ7.37(d,J=8.8Hz,2H),7.21(s,1H),6.82(d,J=8.8Hz,2H),4.14-4.06(m,1H),3.56-3.51(m, 1H),2.38(q,J=7.6Hz,2H),2.17-1.99(m,6H),1.78(d,J=3.1Hz,2H),1.56-1.41(m,3H),1.39-1.08(m,12H),0.84(s,6H). 13 C NMR(101MHz,Chloroform-d)δ156.72,154.42,131.17,121.86,116.60,75.57 ,50.64,48.46,42.72,41.02,32.48,31.04,30.59,30.23,30.21,30.10,9.81.

[0218] Example 52 Synthesis of methyl 4-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)benzoate

[0219] Add cis-4-BOC-aminocyclohexanol (1.00 g, 4.64 mmol), methylparaben (0.7 g, 4.64 mmol), triphenylphosphine (1.8 g, 6.96 mmol), A4 molecular sieve (dried in an oven at 120 °C for 4 h), and THF (10 mL) sequentially to a 100 mL single-necked flask. Purge the solution three times with argon gas, then cool to below -10 °C in an ice-salt bath. Add 5 mL of a THF solution containing DIAD (1.4 g, 6.96 mmol) dropwise at a rate of one drop every two seconds. After 12 hours, monitor the reaction by TLC. Move the flask to 30 °C and add more A4 molecular sieve. After 6 hours, monitor the reaction by TLC again; the reaction is mostly complete, and the reaction is stopped. THF was removed by vacuum concentration, yielding 5.2 g of brownish-yellow oily substance. No solid precipitate was observed after pulping. The sample was packed into a column with 4x silica gel, mixed with 1.2x silica gel, and eluent (EA:PE = 1:10), yielding 0.73 g of white solid product, with a yield of 45.1%.

[0220] Example 53 Synthesis of methyl 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzoate

[0221] Methyl 4-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)benzoate (0.60 g, 2.09 mmol) and DCM (5 mL) were added sequentially to a 100 mL single-necked flask. TFA (4 mL) was then added dropwise at room temperature. After 3 hours, TLC monitoring showed the reaction was complete, and the reaction was stopped. TFA was removed from the reaction solution by vacuum distillation. 0.62 g of crude brown oil was obtained. The crude product was dried in a 60 °C oven for 12 h and used directly in the next step without purification.

[0222] Example 54 Synthesis of methyl 4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoate BTC (0.23 g, 0.79 mmol) and dry DCM (15 mL) were added sequentially to a 100 mL single-necked flask. The flask was cooled to -78 °C in a cold trap. A DCM solution (10 mL) of free p-trifluoromethoxyaniline (0.41 g, 2.32 mmol) and Et3N (1.41 g, 13.92 mmol) was added dropwise over 30 minutes. After the addition was complete, the flask was moved to room temperature and stirred for 4 h.

[0223] A mixed solution of I15·TFA (0.58 g, 1.59 mmol), Et3N (1.41 g, 13.92 mmol), and DCM (10 mL) was added to a 100 mL single-necked flask and reacted at room temperature. The reaction was stopped by TLC monitoring after 2 hours. The sample was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.93 g of crude yellow oil. The sample was packed into a 7x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:5) to give 0.40 g of the product as a white solid, yield 40.0%. ESI-MS: m / z 453.2 [M+H] +

[0224] Example 55 Synthesis of 4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoic acid

[0225] Methyl 4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoate (0.15 g, 0.33 mmol), THF (3 mL), H2O (2 mL), and LiOH (39.8 mg, 0.99 mmol) were added sequentially to a 25 mL single-necked flask. The mixture was stirred at room temperature. After 3 h, TLC monitoring showed that a small portion of the reaction had occurred. The temperature was raised to 30 °C, and 0.1 g of lithium hydroxide was added. After 2 h, TLC monitoring showed that a portion of the reaction had occurred. The temperature was raised to 50 °C, and 0.2 g of lithium hydroxide was added. After 2 h, TLC monitoring showed that the reaction had stopped. The pH was adjusted to 2 with 3N HCl, and the mixture was concentrated under reduced pressure until a small amount of solvent remained. The mixture was filtered, washed with 5 mL of water, and 70 mg of a pale yellow solid was obtained, with a yield of 48.6%.

[0226] Example 56 Synthesis of 4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzamide (SP-C01i)

[0227] At room temperature, add 0.17 g (0.39 mmol) of 4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzoic acid and 10 mL of dry tetrahydrofuran to a 25 mL flask. Then add HATU (0.18 g, 0.46 mmol) and stir for 15 min. DIEA (0.12 g, 0.97 mmol) was added dropwise, and the mixture was stirred for 0.5 h. NH3·H2O (0.3 mL) was added, and after 0.5 h, TLC was performed. The reaction was complete. The mixture was concentrated under reduced pressure to remove tetrahydrofuran. It was extracted twice with dichloromethane (12 mL) and water (10 mL), washed once with saturated brine (25 mL), dried over anhydrous magnesium sulfate, and filtered. The organic phase was concentrated under reduced pressure to obtain 0.20 g of a yellow oily substance. The mixture was packed into a 4x silica gel column, mixed with 1.2x silica gel, and eluted with EA:PE = 1:3. Column chromatography yielded 0.12 g of a pale yellow solid. The yield was 70.5%.

[0228] 1 H NMR(400MHz, DMSO-d6)δ8.81(s,1H),7.82(dd,J=9.4,2.7Hz,3H),7.54-7.45(m,2H),7.21(d,J=8.5Hz,2H),7.14(s,1H),7.02-6.94(m,2H),6.48(d, J=7.6Hz,1H),4.43(tt,J=9.8,4.0Hz,1H),3.54(ddt,J=14.2,10.6,5.3Hz ,1H),2.06(dd,J=12.6,4.3Hz,2H),2.01-1.89(m,2H),1.55-1.30(m,4H). 13 C NMR(101MHz,DMSO-d6)δ167.89,160.26,155.02,142.36,140.48,129.85,12 6.78,122.03,121.97,119.43,118.99,115.28,74.64,47.64,30.44,30.15.

[0229] Example 57 Synthesis of N-(4-(((1R,4r)-4-(3-(((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureoyl)cyclohexyl)oxy)phenyl)propionamide (SP-C01j)

[0230] Add propionic acid (0.02 g, 0.24 mmol, 1 eq), dry tetrahydrofuran (10 mL), HATU (0.034 g, 0.29 mmol, 1.2 eq), and DIEA (0.18 g, 1.74 mmol, 6 eq) to a 25 mL flask and stir for 30 min. Add a THF solution of 1-((1r,4R)-4-(4-aminophenoxy)cyclohexyl)-3-((1r,3R,5S,7R)-3,5-dimethyladamantane-1-yl)urea (0.1 g, 0.24 mmol, 1 eq) in 5 mL. Monitor the reaction by TLC; EA:PE = 1:2. The reaction was complete. Stop the reaction and filter to give 80 mg of white solid. The yield was 71.32%.

[0231] 1 H NMR(400MHz,DMSO-d6)δ9.66(s,1H),7.48(d,J=8.8Hz,2H),6.85(d,J=8.8H z,2H),5.58(d,J=7.5Hz,1H),5.42(s,1H),4.22-4.18(m,1H),2.38-2.31(m, 1H),2.05(s,1H),1.97(d,J=10.5Hz,2H),1.82(d,J=10.5Hz,2H),1.68(s,2 H),1.62-1.55(m,2H),1.07-1.05(m,2H),0.83(t,J=7.4Hz,3H)0.80(s,6H).

[0232] Example 58 Synthesis of N-(4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenyl)acetamide (SP-C01k)

[0233] Add BTC (0.08 g, 0.26 mmol) and dry DCM (10 mL) sequentially to a 50 mL single-necked flask. Cool the flask to -78 °C in a cold trap, and then add dropwise a DCM solution containing 3-fluoro-4-trifluoromethoxyaniline (0.15 g, 0.77 mmol) and Et3N (0.47 g, 4.61 mmol) over 15 minutes. After the addition is complete, bring the flask to room temperature and continue stirring for 0.5 h. After 0.5 h, monitor the solution by TLC; EA:PE = 1:1.

[0234] A mixed solution of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenyl)acetamide·TFA (0.30 g, 0.77 mmol), Et3N (0.47 g, 4.61 mmol), and DCM (5 mL) was added to a 100 mL single-necked flask. The reaction was carried out at room temperature, and the reaction was stopped after TLC monitoring for 0.5 hours. The sample was extracted twice with 15 mL of water, washed once with 15 mL of saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 0.52 g of crude yellow oil. The sample was packed into a 4x silica gel column, mixed with 1.2x silica gel, and eluented with EA:PE = 1:5, EA:PE = 1:3 to give 0.21 g of white solid, with a yield of 36.7%.

[0235] Example 59 Synthesis of ethyl 3-((4-((((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenyl)amino)-3-oxopropionic acid (SP-CO1s)

[0236] Add monoethyl malonate (0.16 g, 0.12 mmol, 1 eq), dry tetrahydrofuran (5 mL), HATU (0.017 g, 0.29 mmol, 1.2 eq), and DIEA (0.09 g, 0.87 mmol, 6 eq) to a 25 mL flask and stir for 30 min. Add a THF (5 mL) solution of 1-((1r, 4r)-4-(4-aminophenoxy)cyclohexyl)-3-(3-fluoro-4-(trifluoromethoxy)phenyl)urea (0.05 g, 0.12 mmol, 1 eq), monitor by TLC (EA:PE = 1:1, AcOH 1 d), the reaction is complete, stop the reaction, and filter to give 42 mg of white solid. The yield is 64.67%. 1 H NMR (400MHz, DMSO-d6) δ9.68 (s, 1H), 8.73 (s, 1H), 7.68 (dd, J = 13.4, 2.5Hz, 1H), 7.50-7.43 (m,2H),7.39(t,J=8.9Hz,1H),7.10(dt,J=9.1,1.8Hz,1H),6.92-6.83(m,2H),6.30(d,J=7 .6Hz,1H),4.24(tt,J=9.6,4.0Hz,1H),3.63-3.46(m,1H),2.27(q,J=7.5Hz,2H),2.02(dd, J=12.7,4.4Hz,2H),1.92(dd,J=12.9,4.0Hz,2H),1.51-1.27(m,4H),1.07(t,J=7.5Hz,3H).

[0237] Example 60 Synthesis of methyl butyrate 4-(4-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)phenoxy)butyrate

[0238] Add tert-butyl ((1r,4r)-4-(4-hydroxyphenoxy)cyclohexyl)carbamate (0.30 g, 0.98 mmol, 1 eq), K₂CO₃ (0.40 g, 2.92 mmol, 3 eq), KI (0.02 g, 0.10 mmol, 0.1 eq), TBAB (0.03 g, 0.10 mmol, 0.1 eq), and then add MeCN (15 mL) to a 25 mL single-necked flask. Add methyl 4-bromobutyrate (0.26 g, 1.46 mmol, 1.5 eq) dropwise, purging with Ar three times, and then heat to reflux. After 3 hours, TLC monitoring showed an EA:PE ratio of 1:3. The reaction was stopped, and the product was concentrated to dryness under reduced pressure. The solution was then extracted with water (20 mL) and DCM (20 mL x 2). The organic layers were combined, washed with water (10 mL), then with saturated brine (10 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain 0.46 g of a brownish-black oily liquid. The crude product was purified by silica gel column chromatography. The column was packed with 1.5 times the volume of sample and 7 times the volume of silica gel, with an EA:PE ratio of 1:15 as the eluent, yielding 0.12 g of a white solid, with a yield of 30.07%.

[0239] Example 61 Synthesis of methyl 4-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenoxy)butyrate

[0240] Methyl 4-(4-(((1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexyl)oxy)phenoxy)butyrate (0.12 g, 0.29 mmol, 1 eq) and DCM (8 mL) were added sequentially to a 100 mL single-necked flask. The mixture was cooled to 0 °C in an ice bath, and TFA (0.5 mL) was added dropwise. After 2 hours, TLC monitoring showed that EA:PE = 1:3, indicating that the reaction was complete, and the reaction was stopped. The DCM and TFA in the reaction solution were removed by vacuum distillation to obtain 0.11 g of crude pale yellow solid. The crude product was used directly in the next step without purification. ESIMS: m / z 308.1 [M+H] + .

[0241] Example 62 Synthesis of 4-(4-(((1R,4r)-4-(3-(((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureoyl)cyclohexyl)oxy)phenoxy)methyl butyrate (SP-C20)

[0242] Add BTC (0.97 g, 3.26 mmol, 0.5 eq) and dry DCM (10 mL) sequentially to a 250 mL single-necked flask. Cool the flask to -80 °C. Add a DCM solution of ammonium phosphate (1.15 g, 6.51 mmol, 1 eq) and Et3N (5.27 g, 52.08 mmol, 8 eq) dropwise over 30 min. After addition, allow the flask to warm to room temperature. After 30 min, monitor the reaction by TLC. If EA:PE = 1:3, the reaction is complete, and the reaction is stopped. The reaction mixture was evaporated to dryness, and DCM (15 mL) and Et3N (5.27 g, 52.08 mmol, 8 eq) were added. The mixture was kept in an ice bath at 0 °C, and a DCM solution (10 mL) of methyl 4-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenoxy)butyrate (2.00 g, 6.51 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed that EA:PE = 1:2, indicating complete reaction, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 4.05 g of a yellow oil. This oil was mixed with 1.5 times the volume of silica gel, packed into a 4 times silica gel column (EA:PE = 1:7), yielding 1.05 g of a white solid. The yield was 31.48%. 1 H NMR (400MHz, DMSO-d6) δ6.94-6.82 (m, 4H), 5.65 (d, J = 7.6Hz, 1H), 5.49 (s, 1H ),4.19(tt,J=10.0,3.9Hz,1H),3.96(t,J=6.3Hz,2H),3.66(s,3H),2.51(t,J =7.3Hz,3H),2.10(p,J=3.2Hz,1H),2.03-1.93(m,4H),1.92-1.83(m,2H),1.7 3(d,J=3.1Hz,2H),1.55(s,4H),1.49-1.15(m,9H),1.13(s,2H),0.85(s,6H). 13 CNMR(101MHz,DMSO-d6)δ173.54,157.00,153.02,151.71,117.66,115.77,75.52,67.31,5 1.79,51.47,50.86,48.60,47.37,42.90,40.99,32.36,30.85,30.61,30.43,30.09,24.77.

[0243] Example 63 Synthesis of 4-(4-(((1R,4r)-4-(3-(((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureoyl)cyclohexyl)oxy)phenoxy)butyric acid (SP-C21)

[0244] SP-C20 (0.50 g, 0.98 mmol, 1 eq), THF (15 mL), H2O (5 mL), and LiOH (0.10 g, 2.93 mmol, 3 eq) were added sequentially to a 25 mL single-necked flask and stirred at room temperature. After 1.5 h, TLC monitoring showed that EA:PE = 1:3, at which point the reaction was stopped. The mixture was concentrated under reduced pressure to remove THF, and extracted with EA (20 mL). The aqueous layer was adjusted to pH 2 with 1 N HCl, and the mixture was filtered under reduced pressure to obtain 0.42 g of a pale yellow solid. The yield was 86.01%. 1 H NMR(400MHz,DMSO-d6)δ6.94-6.74(m,4H),4.16-4.10(m,1H),3.90(t,J=6.4Hz,4H),3.90 (t,J=6.4Hz,2H),3.33(tt,J=10.4,3.8Hz,1H),2.37(t,J=7.3Hz,2H),2.04(dt,J=6.8,3.3 Hz,1H),1.97(d,J=4.1Hz,1H),1.92(dt,J=14.1,6.1Hz,3H),1.82(dd,J=13.5,3.6Hz,2H) ,1.68-1.67(m,2H),1.52-1.44(m,4H),1.41-1.11(m,9H),1.09-1.04(m,2H),0.80(s,6H). 13 C NMR(101MHz,DMSO-d6)δ174.60,157.05,153.08,151.68,117.66,115.79,75.50,67.42,51.47,51.39 ,50.86,48.59,48.55,47.35,42.94,42.90,40.98,32.46,32.36,30.82,30.62,30.39,30.09,24.80.

[0245] Example 64 Synthesis of methyl butyrate (SP-C23) of 4-(4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenoxy)butyrate

[0246] Add BTC (0.03 g, 0.10 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C in a cold trap, then add dropwise a DCM solution of 4-trifluoromethoxyaniline (0.05 g, 0.29 mmol, 1 eq) and Et3N (0.09 g, 0.87 mmol, 3 eq) over 5 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. After 10 minutes, monitor the reaction by TLC. If EA:PE = 1:3, the reaction is complete, and the reaction is stopped. The reaction solution was evaporated to dryness, and DCM (10 mL) and Et3N (0.09 g, 0.87 mmol, 3 eq) were added. The mixture was kept in an ice bath at 0°C, and a DCM solution (5 mL) of methyl 4-(4-(((1r,4r)-4-aminocyclohexyl)oxy)phenoxy)butyrate (0.09 g, 0.29 mmol, 1 eq) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring showed that EA:PE = 1:2, indicating complete reaction, and the reaction was stopped. The DCM was removed by concentration under reduced pressure, yielding 0.15 g of crude white solid. The sample was mixed with 1.5 times silica gel and packed into a 4 times silica gel column (EA:PE = 1:7), yielding 0.10 g of white solid. The yield was 65.28%. 1 H NMR (400MHz, DMSO-d6) δ9.26 (s, 1H), 7.69 (dd, J=13.5, 2.5Hz, 1H), 7.37 (td, J=9.0, 1.2Hz,1H),7.12(ddd,J=9.0,2.6,1.4Hz,1H),6.91-6.79(m,4H),6.77(d,J=7.6Hz, 1H),4.17(tt,J=9.3,3.8Hz,1H),3.91(t,J=6.3Hz,2H),3.61(s,4H),3.57-3.45(m, 1H), 2.46 (t, J = 7.3Hz, 2H), 2.01-1.99 (m, 2H), 1.97-1.89 (m, 4H), 1.44-1.31 (m, 4H). 13 C NMR(101MHz,DMSO-d6)δ173.53,154.85,153.01,152.89,151.67,142.26,142.15,124.63, 117.61,115.77,113.94,106.02,105.78,75.36,51.78,47.73,30.43,30.36,30.29,24.77.

[0247] Example 65 Synthesis of 4-(4-(((1r,4r)-4-(3-(3-fluoro-4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)phenoxy)butyric acid (SP-C24)

[0248] SP-C23 (0.50 g, 0.95 mmol, 1 eq), THF (15 mL), H2O (5 mL), and LiOH (0.10 g, 4.18 mmol, 4.4 eq) were added sequentially to a 25 mL single-necked flask and stirred at room temperature. After 1.5 h, TLC monitoring showed that EA:PE = 1:3, at which point the reaction was stopped. The mixture was concentrated under reduced pressure to remove THF, and then extracted with EA (20 mL). The aqueous layer was adjusted to pH 2 with 1 N HCl, and the mixture was filtered under reduced pressure to obtain 0.47 g of a pale yellow solid. The yield was 96.21%. 1 H NMR (400MHz, DMSO-d6) δ12.11(s,1H),9.19(s,1H),7.68(dd,J=13.5,2.6Hz,1H),7.38 (td,J=8.9,1.2Hz,1H),7.10(ddd,J=9.0,2.6,1.3Hz,1H),6.91-6.79(m,4H),6.57(s, 1H),4.19(tt,J=9.3,3.8Hz,1H),3.90(t,J=6.4Hz,2H),3.54(ddt,J=11.1,7.4,3.9Hz ,1H),2.37(t,J=7.3Hz,2H),2.05-1.97(m,2H),1.96-1.85(m,4H),1.50-1.21(m,5H). 13 C NMR (101MHz, DMSO-d6) δ174.62,155.37,154.80,153.10,152.92,151.65,142.11,142.01,124. 70,117.63,115.81,113.86,105.95,105.71,75.25,67.44,47.62,30.64,30.27,30.13,24.82.

[0249] Example 66 Synthesis of methyl 2-(5-((Z)-4-(((1r,4r)-4-(tert-butoxycarbonyl)amino)cyclohexyl)oxy)benzylidene)-2,4-dioxythiazolidin-3-yl)acetate

[0250] To a 100 mL round-bottom flask, tert-butyl ((1r,4r)-4-(4-formylphenoxy)cyclohexyl)carbamate (4.00 g, 12.53 mmol, 1 eq), methyl 2-(2,4-dioxythiazolin-3-yl)acetate (2.37 g, 12.53 mmol, 1 eq), piperidine (0.50 g, 6.27 mmol, 0.5 eq), acetic acid (0.38 g, 6.27 mmol, 0.5 eq), and toluene (20 mL) were added sequentially, and the mixture was heated to reflux. A solid precipitated as the reaction proceeded. After 8 hours, TLC monitoring showed that EA:PE = 1:1, indicating complete reaction, and the reaction was stopped. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with a small amount of n-hexane to give 3.89 g of a white solid. No product spots were observed in the filtrate on TLC, resulting in a yield of 63.36%.

[0251] Example 67 Synthesis of methyl 2-(5-((Z)-4-(((1r,4r)-4-aminocyclohexyl)oxy)benzyl)-2,4-dioxythiazolidin-3-yl)acetate

[0252] Add methyl 2-(5-((Z)-4-(((1r,4r)-4-(tert-butoxycarbonyl)amino)cyclohexyl)oxy)benzyl)-2,4-dioxythiazolidin-3-yl)acetate (3.89 g, 7.94 mmol, 1 eq) and DCM (8 mL) sequentially to a 100 mL single-necked flask. Cool to 0 °C in an ice bath, and add TFA (2 mL) dropwise. After 4 hours, TLC monitoring showed that EA:PE = 1:3, indicating complete reaction, and the reaction was stopped. Remove DCM and TFA from the reaction solution by vacuum distillation to obtain 3.87 g of crude pale yellow solid. The crude product was used directly in the next step without purification. ESIMS: m / z 390.1 [M+H] + .

[0253] Example 68 Synthesis of methyl acetate 2-(2,4-dioxo-5-((Z)-4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzyl)thiazolidin-3-yl)acetate (SP-D01)

[0254] Add BTC (0.03 g, 0.10 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C. Add 10 mL of a DCM solution containing 4-trifluoromethoxyaniline (0.05 g, 0.29 mmol, 1 eq) and Et3N (0.13 g, 1.30 mmol, 5 eq) dropwise over 5 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. After 10 minutes, monitor the reaction by TLC. If EA:PE = 1:3, the reaction is complete, and the reaction is stopped. The reaction solution was evaporated to dryness, and DCM (10 mL) and Et3N (0.13 g, 1.30 mmol, 5 eq) were added. The mixture was kept in an ice bath at 0 °C, and a solution of methyl 2-(5-((Z)-4-(((1r,4r)-4-aminocyclohexyl)oxy)benzylene)-2,4-dioxythiazolidin-3-yl)acetate (0.10 g, 0.26 mmol, 1 eq) in DCM (5 mL) was added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature. TLC monitoring was performed after 30 min. The reaction was complete after EA:PE = 1:1 and AcOH was added for 2 days. The reaction was then stopped. The DCM was removed by concentration under reduced pressure, yielding 0.23 g of crude white solid. The crude product was purified by silica gel column chromatography. The column was packed with 1.5 times the volume of the sample and 5 times the volume of silica gel. The eluent was EA:PE = 1:10, yielding 0.10 g of white solid, with a yield of 66.67%. ESIMS: m / z 616.0 [M+Na] + .1H NMR (400MHz, DMSO-d6) δ9.74(s,1H),7.96(s,1H),7.65-7.57(m,2H),7.54-7.45(m,2H),7.22(d,J=8.6Hz,2H),7.18-7.12(m,2H),6.44(d ,J=7.6Hz,1H),4.51(s,2H),3.72(s,3H),3.57-3.53(m,1H),3.28-3.25(m,1H),2.12-2.03(m,2H),1.98-1.90(m,2H),1.58-1.32(m,6H).

[0255] Example 69 Synthesis of 2-(2,4-dioxo-5-((Z)-4-(((1r,4r)-4-(3-(4-(trifluoromethoxy)phenyl)ureo)cyclohexyl)oxy)benzyl)thiazolidin-3-yl)acetic acid

[0256] SP-D01 (0.08 g, 0.13 mmol, 1 eq), THF (5 mL), H2O (1 mL), and LiOH (0.01 g, 0.40 mmol, 3 eq) were added sequentially to a 25 mL single-necked flask and stirred at room temperature. After 1.5 h, TLC monitoring showed that EA:PE = 1:3, at which point the reaction was stopped. The mixture was concentrated under reduced pressure to remove THF, and the pH was adjusted to 4 with 1N HCl. The mixture was then extracted with EA (10 mL) and H2O (10 mL). The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to give 70 mg of a white solid. Yield: 92.98%. ESIMS: m / z 578.3 [MH] - .

[0257] Example 70 Synthesis of methyl acetate 2-(5-((Z)-4-(((1R,4r)-4-(3-((1R,3R,5S,7R)-3,5-dimethyladamantane-1-yl)ureoyl)cyclohexyl)oxy)benzylidene)-2,4-dioxythiazoline-3-yl)acetate

[0258] Add BTC (0.03 g, 0.10 mmol, 0.34 eq) and dry DCM (5 mL) sequentially to a 25 mL single-necked flask. Cool the flask to -80 °C in a cold trap. Add a 10 mL solution of DCM containing memantine (0.05 g, 0.28 mmol, 1 eq) and Et3N (0.28 g, 2.80 mmol, 10 eq) dropwise over 5 minutes. After the addition is complete, allow the flask to warm to room temperature naturally. After 10 minutes, monitor the reaction by TLC. If the EA:PE ratio is 1:3, the reaction is complete, and the reaction is stopped. Evaporate the reaction solution to dryness. Add DCM (10 mL) and Et3N (0.28 g, 2.80 mmol, 10 eq). Cool the flask to 0 °C in an ice bath. Add a 5 mL solution of DCM containing N4 (0.10 g, 0.28 mmol, 1 eq) dropwise. After the addition is complete, allow the flask to react at room temperature. After 30 minutes, monitor the reaction by TLC. If the EA:PE ratio is 1:1 and the reaction proceeds with AcOH for 2 days, the reaction is complete, and the reaction is stopped. DCM was removed by vacuum concentration, yielding 0.31 g of crude white solid. The crude product was purified by silica gel column chromatography: 1.5 times the volume of sample was mixed, and the column was packed with 5 times the volume of silica gel. The eluent was EA:PE = 1:10, yielding 0.08 g of white solid, with a yield of 48.20%. ESIMS: m / z 594.66 [MH] - . 1H NMR(400MHz,DMSO-d6)δ7.96(s,1H),7.65-7.56(m,2H),7.18-7.09(m,2H), 5.62(d,J=7.6Hz,1H),5.43(s,1H),4.51(s,2H),4.45(dd,J=8.9,4.7Hz,1H) ,3.72(s,3H),2.09-1.99(m,3H),1.89-1.81(m,2H),1.68(d,J=3.1Hz,2H),1 .50(s,4H),1.48-1.37(m,2H),1.33-1.14(m,7H),1.08(s,2H),0.80(s,6H).

[0259] Test Example 1

[0260] 1. sEH inhibitory activity test

[0261] Detection Principle: The specific substrate (3-phenyl-oxy)-cyanoacetate-(6-methoxy-naphth-2-yl)methyl ester, i.e., PHOME, is itself non-fluorescent. However, under the action of sEH enzyme, it is hydrolyzed to produce the product 6-methoxy-2-naphthaldehyde. 6-methoxy-2-naphthaldehyde can emit fluorescence at a wavelength of 465 nm when excited by light at 330 nm. The intensity of the detected fluorescence signal is inversely proportional to the strength of the inhibitory effect on sEH enzyme. Based on the above principle, the inhibition rate of samples at different concentrations was calculated compared with the positive control group. The IC50 of the compound was calculated using SPSS 20 software based on the inhibition rate and concentration. 50 value.

[0262] 2. Preparation of reagents and drugs

[0263] 25mM Tris-HCl buffer (pH=7.4, containing 0.1mg / mL BSA): Take 12.5mL of 1M Tris-HCl buffer, add 5mg BSA, dilute with purified water and adjust the pH to 7.4 with hydrochloric acid, and bring the volume to 500mL.

[0264] PHOME solution: Dissolve 0.79 mg of PHOME in 106 μL of DMSO to obtain a 20 mM PHOME solution. Dilute to 1 / 3 mM with Tris-HCl buffer before use.

[0265] sEH solution: The sEH (5 mg / mL) stock solution was stored at -80°C and diluted to 4 μg / mL with 25 mM Tris-HCl buffer before use.

[0266] The sample powder to be tested is dissolved in DMSO to prepare a 20mM solution, stored at -20℃ for later use, and diluted with Tris-HCl buffer to the corresponding concentration before use.

[0267] 3. Experimental Grouping

[0268] Experimental design: solvent group, 100% activity group (A), inhibitor group (B), positive control group (C), as shown in Table 1.

[0269] Table 1 Experimental Grouping

[0270] hole buffer solution DMSO Inhibitors sEH Substrate Solvent group 168μL 2μL — — 30μL 100% Vitality Group (A) 148μL 2μL — 20μL 30μL Inhibitor group (B) 148μL — 2μL 20μL 30μL Positive control group (C) 148μL — 2μL 20μL 30μL

[0271] 4. Experimental Procedure

[0272] (a) Add 148 μL / well of Tris-HCl buffer to a 96-well black microplate;

[0273] (b) Add 2 μL of the test sample solution. Replace the solvent group and the 100% activity group with an equal volume of DMSO. Add the lead compound GL-B401 (structural formula: [insert structural formula here]) to the positive control group.

[0274] (c) The inhibitor group had a total of 5 concentrations, with final concentrations of 10 nM, 5 nM, 2.5 nM, 1.25 nM and 0.625 nM, respectively;

[0275] (d) Add 20 μL of s-EH solution (final concentration of 400 ng / mL), and replace the solvent group with an equal volume of Tris-HCl buffer;

[0276] (e) Add 30 μL of PHOME substrate to start the reaction (final concentration 50 μM) and incubate at 37 °C for 10 min;

[0277] (f) ELISA reader detects fluorescence signal data, with an excitation wavelength of 330 nm and an emission wavelength of 465 nm.

[0278] 5. Data Analysis

[0279] Each sample was prepared in triplicate, and the mean value of the three replicates was the fluorescence value (F) of the analyte. The inhibition rate % was calculated as [(AF - BF) / AF] × 100, where AF is the fluorescence value of the 100% activity group and BF is the fluorescence value of the inhibitor group. The IC50 of the compound was calculated using SPSS 20 software based on the inhibition rate and concentration. 50 value.

[0280] This invention employs a biochemical method, using cyano(6-methoxynaphthyl-2-yl)2-(3-phenylethyleneoxy-2-yl)acetic acid methyl ester (PHOME) as a substrate, and evaluates the activity of the compounds using recombinant human sEH (HsEH) and murine sEH (MsEH). The results are shown in Table 2.

[0281] Table 2. Human(HsEH) and Murine(MsEH)sEH IC of the compounds 50

[0282]

[0283]

[0284] Test Example 2

[0285] 1. PPARγ agonist activity assay

[0286] Detection principle: When pM-hPPAR binds to an appropriate ligand, it is activated and binds to the GAL4 DNA binding site in plasmid pB4-RES-tk-luc, thereby initiating the expression of the downstream luciferase reporter gene. The presence of luciferase is used to determine whether the test compound is an agonist of PPAR.

[0287] 2. Test Results: SP-C01 for PPARγEC 50 =4.2μM, SP-A01 for PPARγEC 50 =6.77μM, SP-A07 for PPARγEC 50 =7.75μM, SP-B07 for PPARγEC 50 =1.43μM.

[0288] Table 3. PPARγ of the compounds in the examples

[0289]

[0290]

[0291] Test Example 3: Acute Toxicity Test

[0292] 3.1 Laboratory Animals

[0293] Healthy KM mice, half male and half female, SPF grade, 18-22g, were housed in an environment with a temperature of 24±2℃, relative humidity of 60±10%, and alternating 12h light and 12h dark conditions, with free access to food and water.

[0294] 3.2 Experimental Reagents and Chemicals

[0295] Table 4 Laboratory Supplies

[0296] name Manufacturer batch number SP-B07 — / physiological saline Nanjing Senbeijia Biotechnology Co., Ltd. PM20080903 DMSO Sigma 20180923 Twain 80 Sinopharm Chemical Reagent Co., Ltd. 20121225

[0297] 3.3 Experimental Methods

[0298] (1) Drug preparation

[0299] Accurately weigh SP-B07 and SP-C01 and dissolve them in a mixed solvent of DMSO and Tween 80 (1:1). After they are completely dissolved, add 9 times the volume of physiological saline to the mixed solvent for dilution, and finally obtain a drug solution of 1 mg / mL as the dosage.

[0300] (2) Grouping, feeding and administration

[0301] After one week of acclimatization feeding, male and female mice were randomly assigned to 5 groups (n=8) according to body weight: normal control group (Vehicle), SP-B07 1g / kg group, and SP-C01 1g / kg group. Each group was administered the drug by gavage at a volume of 10mL / kg for 14 consecutive days. The other two groups of mice were given SP-C01 1g / kg and SP-C01 5g / kg respectively as a single dose and were observed for 14 consecutive days.

[0302] (3) Physiological condition and weight changes of mice

[0303] Immediately after administration, mice were observed for 30 minutes to monitor for adverse reactions or death. Simultaneously, observations were conducted at 1 hour, 2 hours, 4 hours, and 8 hours to monitor for adverse reactions or death. Subsequently, the mice's daily activity and mental state were monitored and recorded daily, and the weight of each group of mice was measured and recorded weekly.

[0304] (4) Detection of organ indices in mice

[0305] Two weeks after drug administration, mice in each group underwent gross dissection. The heart, liver, spleen, lungs, and kidneys of each mouse were removed, rinsed with physiological saline, and dried with medical gauze before being placed in EP tubes for later analysis. Organ indices were calculated using the following formula:

[0306] Organ Index (%) = Organ Weight / Body Weight × 10%

[0307] 3.4 Data Statistics

[0308] Experimental data are expressed as mean ± standard error (Mean ± SEM) and statistically analyzed using GraphPad Prism 8.4 software. One-way ANOVA was used for comparisons among multiple groups, and LSD method was used for homogeneity of variance. P < 0.05 was considered statistically significant.

[0309] 3.5 Experimental Results

[0310] Effects of compounds on organ indices in male and female mice, such as Figure 2 , Figure 3 As shown, there were no significant differences in visceral indices between the drug-treated group and the control group. Figure 2The effect of compound SP-C01 on body weight in KM mice was investigated. SP-C01, prepared in 0.5% CMC Na solution, was administered by gavage at doses of 1.0 g / kg and 5.0 g / kg, respectively. The control group was given 0.5% CMC Na. Figure 2 In the study, a single dose (op) of SP-C01 1.0 g / kg and 5.0 g / kg with 0.5% CMC Na was administered and body weight was monitored for 14 days. Figure 2 The patients were continuously administered SP-C01 1.0 g / kg and 0.5% CMC Na for 14 days, and their body weight was monitored. Figure 3 The effect of compound SP-C01 on the visceral index of KM mice was investigated. SP-C01, prepared in 0.5% CMC Na solution, was administered by gavage at doses of 1.0 g / kg and 5.0 g / kg, respectively. The control group was given 0.5% CMC Na. Figure 3 In this study, SP-C01 was administered as a single dose (op) at 1.0 g / kg, 5.0 g / kg, and 0.5% CMC Na, and visceral indices were calculated on day 14. Figure 3 In this study, patients were continuously administered SP-C01 1.0 g / kg and 0.5% CMC Na for 14 days, and visceral index was calculated on day 14.

[0311] Figure 4 These are representative H&E stained sections showing the effects of compound SP-C01 on organs in KM mice. SP-C01, prepared in 0.5% CMC Na solution, was orally administered at a dose of 1.0 g / kg for 14 consecutive days. The control group was given 0.5% CMC sodium. No significant abnormalities were observed. After 14 days, no significant differences were observed between the treated group and the control group. Further pathological examination of heart, liver, spleen, lung, kidney, and stomach tissues showed no significant differences compared to the control group.

[0312] Test Example 5: Liver Microsomal Stability

[0313] The microsomal stability of compounds SP-B07 and SP-C01 was evaluated in the liver microsomes of SD rats. The tested compounds showed satisfactory stability in vitro in microsomal stability. Compound SP-B07, in rat liver microsomes, had a half-life (t...) 1 / 2 The half-life (t) of compound SP-C01 in rat liver microsomes is 5.50 h. 1 / 2 The time to metabolism was 5.75 h. This preliminary result indicates that compounds SP-B07 and SP-C01 are metabolically stable and have potential for further development.

[0314] Table 5. Average concentration (ng / mL) of SP-B07 in microsomal buffer at different times.

[0315]

[0316] Table 6. Average concentration (ng / mL) of SP-CO1 in microsomal buffer at different times.

[0317]

[0318] Test Example 6: Plasma Protein Binding Rate (%PPB)

[0319] Plasma protein binding (%PPB) in SD rats was measured using a classic balanced dialysis apparatus. Compound SP-B07 showed a moderate %PPB (95.29%). Compound SP-C01 showed a moderate %PPB (83.9%). Since the therapeutic effect of a drug depends on the concentration of the free drug, a moderate %PPB concentration may be beneficial for in vivo efficacy, thereby producing analgesic and anti-inflammatory effects.

[0320] Test Example 7 Pharmacokinetics

[0321] To determine the metabolic stability of compounds SP-B07 and SP-C01 in vivo, pharmacokinetic characteristics were determined in 6-8 week old Sprague-Dawley (SD) rats (equal numbers of males and females) by oral (ig) single dose of 50 mg / kg and intravenous (iv) single dose of 10 mg / kg. Pharmacokinetic parameters are shown in Table 7. Blood levels were measured within 8 hours after oral administration. The maximum concentration reached after 0.5 h (1.14 μM); the area under the curve (AUC)... 0-8h The plasma t of SP-B07 was 13.80 (μM·h); 1 / 2 The time to peak concentration (3.29 μM) was 1.63 h. The maximum concentration was reached 2 min after intravenous injection of compound SP-B07; the area under the curve (AUC) was 1.63 h. 0-8h The plasma t of SP-B07 was 14.12 μM·h. 1 / 2 The time to bioavailability was 1.91 h. SP-B07 has a moderate bioavailability of 19.54%, laying a good foundation for further in vivo studies. The maximum concentration (20.71 μM) reached after 10.5 h was SP-C07; the area under the curve (AUC) was [missing value]. 0-8h The plasma t of SP-C01 was 38.88 (μM·h); 1 / 2 The time to peak concentration (5.71 μM) was 1.63 h. The maximum concentration was reached 2 min after intravenous injection of compound SP-C01; the area under the curve (AUC) was 1.63 h. 0-8h The plasma concentration of SP-C01 was 10.72 μM·h. 1 / 2 The duration of action was 1.91 hours. SP-C01 showed a high bioavailability of 72.54% (as shown in Table 8). This lays a good foundation for further in vivo studies.

[0322] Table 7. Pharmacokinetics of SP-B07 in rats after intravenous and oral administration (n=3).

[0323]

[0324] Table 8. Pharmacokinetics of SP-C01 in rats after intravenous and oral administration (n=3).

[0325]

[0326] Test Example 7: Freund's Complete Adjuvant (CFA) Induced Arthritis Mouse Model (AIA)

[0327] Freund's complete adjuvant (CFA) induced arthritis mouse model (AIA) has similar characteristics to human rheumatoid arthritis (RA) in terms of pathogenesis, joint pain, bone destruction, and synovial hyperplasia, and is now widely used in research on the pathogenesis and treatment mechanisms of human RA.

[0328] To determine the effectiveness of compound SP-C01 in treating AIA in female KM mice (20-22g), this invention designed five groups for comparative intraperitoneal injection: the model group (CFA), the SP-C01 group (SP-C01 - 5mg / kg), the SP-C01 group (SP-C01 - 10mg / kg), the SP-C01 group (SP-C01 - 20mg / kg), and the celecoxib group (celecoxib - 10mg / kg). First, this invention used the hot plate method to detect the pain threshold of AIA mice, measuring the percentage increase in pain threshold at 10, 45, 120, and 240 minutes after drug administration. Figure 5 (A), and plotted the curve of pain threshold increasing over time ( Figure 5 From B), the AUC value of the percentage increase in pain threshold was obtained. Figure 5 (C). Compound SP-C01 began to raise the pain threshold 10 minutes after administration and remained at a high level until 4 hours. Celecoxib raised the pain threshold within 2 hours, and its effect began to decrease after 4 hours. The curve showing the percentage increase in pain threshold indicates that the analgesic effect of compound SP-C01-5 mg / kg was superior to that of celecoxib-10 mg / kg after 60 minutes, and SP-C01-10 mg / kg was more effective than celecoxib-10 mg / kg, demonstrating a clear dose-response relationship. Finally, this invention evaluated the paw thickness of AIA mouse models at different time points after compound injection and compared it with the paw thickness before modeling, calculating the paw swelling reduction rate. (Figure...) Figure 5The DF analysis showed that compound SP-C01 significantly reduced swelling in mouse paws, and was superior to celecoxib. In conclusion, compound SP-C01 has significant analgesic and anti-inflammatory effects (reducing swelling in mouse paws) on (CFA)-induced AIA, and is significantly superior to celecoxib-10 mg / kg.

[0329] Figure 5 To evaluate the effect of compound SP-C01 on the treatment of CFA-induced arthritis in a mouse model (AIA); 24 hours after inducing CFA with AAI (25 μL / 25 g, ip), SP-C01 (5 mg / kg, ip, ip) was administered. Figure 5 In this context, A represents the percentage increase in pain threshold among the CFA group, CFA+SP-C01 group, and CFA+celecoxib group. Figure 5 B in the diagram represents the trend of the percentage of pain threshold over time in the CFA group, CFA+SP-C01 group, and CFA+celecoxib group within 240 minutes. Figure 5 In this context, C represents the area under the curve of the percentage increase in pain threshold in the CFA group, CFA+SP-C01 group, and CFA+celecoxib group. Figure 5 D in the figure represents the swelling reduction rate of the CFA group, the CFA+SP-C01 group, and the CFA+celecoxib group; Figure 5 E in the figure represents the trend of the percentage of swelling reduction within 12 hours in the CFA group, CFA+SP-C01 group and CFA+celecoxib group over time. Figure 5 F in the figure represents the area under the curve of the swelling reduction rate within 12 hours in the CFA group, CFA+SP-C01 group, and CFA+celecoxib group; significance: compared with the Mod group, ****P<0.0001, ***P<0.001, **P<0.01 and *P<0.05, and compared with the celecoxib group, ##P<0.01 and #P<0.05.

[0330] Six groups were then designed for comparison via gavage: the model group (CFA), the low-dose SP-C01 group (SP-C01-30mg / kg), the high-dose SP-C01 group (SP-C01-60mg / kg), the low-dose SP-C01h group (SP-C01h-30mg / kg), the high-dose SP-C01h group (SP-C01h-60mg / kg), and the celecoxib group (celecoxib-30mg / kg). First, this invention used complete Freund's adjuvant (CFA) (25μL / 25g) to inject the right posterior metatarsal region of female C57BL / 6 mice (20-22g). Twenty-four hours after modeling, the pain threshold of mice with AIA was measured using the hot plate method. The percentage increase in pain threshold was measured at 10, 30, 45, 60, 120, and 240 minutes after drug administration. Figure 6(A) and plotted the curve of pain threshold increasing over time ( Figure 6 In B), the AUC value of the percentage increase in pain threshold was obtained. Figure 6 (C) Compound SP-C01 began to raise the pain threshold 10 minutes after administration and remained at a high level until 4 hours. Celecoxib raised the pain threshold 30 minutes after administration, slightly later than SP-C01, and its effect began to decrease at 4 hours. The curve showing the percentage increase in pain threshold indicates that compound SP-C01 (30 mg / kg) has a better analgesic effect than celecoxib (30 mg / kg). Finally, this invention evaluated the paw thickness in AIA mouse models at different time points after compound injection and compared it with the paw thickness before modeling. The paw swelling reduction rate was calculated. (See figure...) Figure 6 The DF data shows that compound SP-C01 significantly reduced swelling in mouse paws, and was significantly better than celecoxib. In conclusion, compound SP-C01, whether administered intraperitoneally or by gavage, has significant effects on analgesia and anti-inflammation (reducing swelling in mouse paws) induced by cerecognition of fatty acids (CAA), and is significantly superior to celecoxib.

[0331] Figure 6 To evaluate the effect of compound SP-C01 on the treatment of CFA-induced arthritis in a mouse model (AIA); 24 hours after inducing CFA with AAI (25 μL / 25 g, ip), C57BL / 6 mice were administered SP-C01 (30 mg / kg, ig), SP-C01, SP-C01h (30 mg / kg, ig), and celecoxib (30 mg / kg); Figure 6 (A) represents the percentage increase in pain threshold in the CFA group, CFA+SP-C01 group, CFA+SP-C01h group, and CFA+celecoxib group; Figure 6 B in the diagram represents the trend of the percentage increase in pain threshold over time within 240 minutes for the CFA group, CFA+SP-C01 group, CFA+SP-C01h group, and CFA+celecoxib group. Figure 6 In this context, C represents the area under the curve for the percentage increase in pain threshold among the CFA group, CFA+SP-C01 group, CFA+SP-C01h group, and CFA+celecoxib group. Figure 6 D in the figure represents the swelling reduction rate of the CFA group, CFA+SP-C01 group, CFA+SP-C01h group, and CFA+celecoxib group; Figure 6 E in the figure represents the trend of the percentage of swelling reduction within 12 hours in the CFA group, CFA+SP-C01 group, CFA+SP-C01h group and CFA+celecoxib group over time. Figure 6F in the figure represents the area under the curve of the swelling reduction rate within 12 hours for the CFA group, CFA+SP-C01 group, CFA+SP-C01h group, and CFA+celecoxib group; Significance: Compared with the Mod group, ***P<0.0001, ***P<0.001, **P<0.01 and *P<0.05; compared with the celecoxib group, ##P<0.01 and #P<0.05; compared with the Mod group, $$P<0.001, $$P<0.01 and $P<0.05.

[0332] Test Case 9: Neuropathic pain induced by chronic contraction injury of the sciatic nerve (CCI)

[0333] The effects of SP-C01 were further investigated in a neuropathic pain model induced by chronic contraction injury (CCI) of the sciatic nerve in SD rats. Rats were randomly assigned to oral (±)-EC-5026 (30 mg / kg / day) and SP-C01 groups. The SP-C01 group was further divided into 30 mg / kg / day and 100 mg / kg / day groups, with an additional intraperitoneal injection group of SP-C01 30 mg / kg / day. After oral administration of (±)-EC-5026 (30 mg / kg / day) and SP-C01 (30 mg / kg / day) at 12-hour dosing intervals (BID), pain relief was assessed in rats on days 1, 3, and 7 throughout the study using the von Frey test. Pain relief throughout the study was assessed by an increase in the mechanical claw withdrawal threshold (PWT). The test showed that SP-C01 had no significant tolerance and had a certain analgesic effect (Figure 4A-B). Following oral administration of SP-C01 (30 mg / kg / day), SP-C01 (100 mg / kg / day), and intraperitoneal injection of SP-C01 (30 mg / kg / day) at 12-hour dosing intervals (BID), the mechanical claw withdrawal threshold (PWT) was assessed using a von Frey test during the first 7 hours of the study. The efficacy against neuropathic pain increased in a dose-dependent manner, with oral administration showing roughly the same efficacy as intraperitoneal injection. Figure 7 Experimental results showed that the overall analgesic effect of SP-C01 was cumulative over time, and the therapeutic effect was significantly better than that of the positive control drug EC5026 from the 7th day.

[0334] Figure 7 In a chronic contractile injury model of neuropathy in male SD rats, SP-C01 blocked pain as measured by the von Frey test (mechanical withdrawal threshold). Data are presented as mean ± standard error of mean. SP-C01, formulated in 10% DMSO and 90% corn oil, was administered orally at a specified dose at 12-hour dosing intervals (BID). (n = 8 per group); Figure 7In this context, A represents oral administration of (±)-EC-5026 (30 mg / kg / day) and SP-C01 (30 mg / kg / day) to assess pain relief over the first 7 hours. Figure 7 B in the text refers to the oral administration of (±)-EC-5026 (30 mg / kg / day) and SP-C01 (30 mg / kg / day) throughout the study to assess pain relief on days 1, 3, and 7. Figure 7 The C in the formula refers to oral SP-C01 (30 mg / kg / day) and SP-C01 (100 mg / kg / day) and SP-C01 intraperitoneal injection (30 mg / kg / day) used to evaluate the dose-dependent pain relief of SP-C01 and the effect of SP-C01 on different administration methods, characterized by an increase in the mechanical gripper contraction threshold (PWT). Figure 7 D in the table represents the sum of AUC values ​​for all experimental results. Significance: Compared with the Mod group, ***P<0.01, **P<0.01, and *P<0.05;

[0335] Test Example 10: L-arginine-induced acute pancreatitis model

[0336] Acute pancreatitis is a potentially life-threatening gastrointestinal disease with an increasing incidence over the past few decades. Currently, there are no effective treatments, making drug research for acute pancreatitis extremely urgent. Given the association between sEH and the pathogenesis of acute pancreatitis, this study evaluated the efficacy of the sEH inhibitor SP-C01 at doses of 5 mg / kg and 10 mg / kg in treating an L-arginine-induced acute pancreatitis model in male C57BL / 6 mice (20-22 g), comparing it with celecoxib (5 mg / kg) and ulinastatin (5 mg / kg). Histological analysis of the pancreas was performed to determine whether SP-C01 treatment reduced the severity of L-arginine-induced pancreatitis. Pathological changes were examined on H&E-stained pancreatic sections. Figure 8 ).

[0337] Figure 8 Histological analysis of the pancreas in mice treated with control, Mod, celecoxib, ulinastatin, and compound SP-C01. Representative H&E stained sections of the pancreas from the in vivo efficacy studies. Figure 9 The results of histological analysis of the pancreas of mice treated with control, Mod, celecoxib, ulinastatin, and compound SP-C01. (A) Edema (B) Inflammatory cells (monocytes and polymorphonuclear cells) (C) Parenchymal atrophy (D) Total score (edema, mononuclear, polymorphonuclear, and parenchymal atrophy) (n=5 per group) *p<0.05, **p<0.01 vs Mod.

[0338] As expected, the L-arginine model group (5.75 ± 1.64) showed pancreatic damage representative of AP, including edema ( Figure 9 A) Inflammatory cell infiltration ( Figure 9 B) and solid atrophy ( Figure 9 (C in the text). In comparison, both SP-C01 5 mg / kg (2.40±1.20) and SP-C01 10 mg / kg (1.60±0.49) improved L-arginine-induced pancreatic injury in acute pancreatitis (AP). The dose of compound SP-C01 10 mg / kg was superior to the dose of 5 mg / kg, and compound SP-C01 5 mg / kg (2.40±1.20) was more effective than celecoxib 5 mg / kg (3.50±1.80) in reversing pancreatic injury, edema, and neutrophil infiltration. Compound SP-C01 10 mg / kg (1.60±0.49) was more effective than ulinastatin 5 mg / kg (2.00±0.71) in reversing pancreatic injury and edema.

[0339] Test Example 11: A diabetic model induced by CD60 diet combined with streptozotocin

[0340] Diabetes mellitus is a chronic metabolic disease characterized primarily by elevated blood glucose levels, and its incidence has been increasing over the past few decades. Currently, there are no effective treatments, making drug research for treating diabetes and its related complications, such as diabetic pain, extremely urgent. Given the association between sEH and PPAR with the pathogenesis of diabetes and related complications, this study evaluated the effects of sEH inhibitors SP-C01 and SP-B07 at doses of 30 mg / kg and 90 mg / kg on CD60-positive diets combined with streptozotocin-induced diabetes and diabetic nephropathy, compared with pioglitazone (3 mg / kg), in male C57BL / 6 mice (20–22 g). Glucose analysis was performed to determine whether SP-C01 and SP-B07 treatment reduced the severity of diabetes and related complications. Figures 10-11 ).

[0341] Figure 10 The results show the blood glucose analysis of mice treated with control, Mod, pioglitazone, compound SP-C01, and compound SP-B07. Figure 11 To use the hot plate method respectively ( Figure 11 (A and B in the text) and Von-Frey mechanical stimulation ( Figure 11 The treatment outcome of diabetic neuropathic pain, measured in group C), was as follows. Compared with the control group, ****P<0.0001, ***P<0.001, **P<0.01 and *P<0.05.

[0342] As expected, the model group exhibited diabetic complications such as hair loss, increased periorbital discharge, and limited mobility, while the treatment group showed no significant behavioral abnormalities compared to healthy mice. Both SP-C01 and SP-B07 effectively lowered blood sugar and achieved the improvement of diabetic neuropathic pain not found in the PPARγ single-target agonist pioglitazone.

[0343] Test Example 12: Effects of oral SP-CO1 on a flurryl adjuvant-induced mouse model of rheumatoid arthritis and a rat model of chronic granulomatosis.

[0344] Seventy-two qualified female ICR mice, weighing 19–26 g, were randomly divided into four groups according to body weight: a normal control group, a model control group, a celecoxib group, a low-dose SP-CO1 group, a medium-dose SP-CO1 group, and a high-dose SP-CO1 group, with ten animals in each group. Except for the normal control group, each of the other groups received an injection of 25 μL of Fluoride complete adjuvant per animal into the right paw. Before administration, different drugs were prepared to the corresponding concentrations using pure water. Thirty minutes after modeling, each group was administered the different test drug orally at a dose of 20 mL / kg once a day. The normal control group and the model control group received the same volume of pure water. All groups were administered the drug before the first administration (before modeling). Pain threshold screening was performed, and the pain threshold of the foot was measured using a hot plate tester at 30 min, 60 min, 120 min, and 240 min after the first administration. The pain threshold improvement rate (%) was calculated as follows: pain threshold improvement rate (%) = (pain threshold after administration - pain threshold before administration) / pain threshold before administration × 100%. The thickness of the right foot was also measured at 1, 2, 4, 6, 8, and 12 h after administration. On the 20th day after modeling, all model animals were regrouped according to their joint index scores and administered the drug by gavage at a dose of 20 mL / kg once a day for 7 consecutive days. The joint index scores of each group were measured on the 3rd and 7th days after administration, and the pain threshold of the animals was measured using a hot plate tester after the last administration.

[0345] Table 9 Group and Dosage Design

[0346] Group Dosage (mg / kg) Volume (mL / kg) Concentration (mg / mL) Number of animals (individuals) normal control group － 20 － 10 Model control group － 20 － 10 Celecoxib group 30 20 1.5 10 SP-C01 low-dose group 15 20 0.75 10 SP-C01 medium dose group 30 20 1.5 10 SP-C01 high-dose group 60 20 3 10

[0347] Effects of SP-C01 on chronic granuloma in rats

[0348] Fifty qualified SD rats (half male and half female), weighing 210–260 g, were randomly divided into four groups (n=10 per group): a model control group, a celecoxib group, a low-dose SP-C01 group, a medium-dose SP-C01 group, and a high-dose SP-C01 group. After isoflurane inhalation anesthesia, approximately 50 mg cotton balls were implanted in both groins of the rats. Strict aseptic technique was maintained during the procedure. Postoperatively, penicillin was injected intraperitoneally for anti-inflammatory purposes. Before administration, different drugs were prepared to a concentration of 0.5 mg / mL using pure water and administered orally at a dose of 10 mL / kg to each group for 7 consecutive days. The model control group received an equal volume of pure water. After the last administration, the animals were anesthetized with isoflurane inhalation, bled, and the cotton balls containing granulomas were removed and weighed. The degree of swelling was calculated as (wet weight of the cotton ball granuloma - dry weight of the cotton ball).

[0349] Table 10 Group and Dosage Design

[0350] Group Dosage (mg / kg) Volume (mL / kg) Concentration (mg / mL) Number of animals (individuals) Model control group － 10 － 10 Celecoxib group 20 10 2 10 SP-C01 low-dose group 10 10 1 10 SP-C01 medium-dose group 20 10 2 10 SP-C01 high-dose group 40 10 4 10

[0351] Data processing and statistical analysis

[0352] The data in this experiment were rounded to the nearest whole number and statistical analysis was performed according to the standard operating procedure (SOP). SPSS 22.0 was used for statistical analysis. Quantitative data are expressed as mean ± standard deviation. The results indicate that Leven's test was used to test for normality and homogeneity of variance. If there was no statistical significance (P>0.05), one-way ANOVA was used for statistical analysis. If the ANOVA was statistically significant (P≤0.05), the LSD test (parametric method) was used for comparative analysis. If the variances were unequal (P≤0.05), the Kruskal-Wallis test was used. If the Kruskal-Wallis test was statistically significant (P≤0.05), Dunnett's test (nonparametric method) was used for comparative analysis. The statistical results were presented with α = 0.05 as the test limit, where P≤0.05 indicated statistical significance, and P≤0.01 indicated a highly significant difference.

[0353] Experimental results

[0354] Effects of SP-C01 on a Fluoride complete adjuvant-induced mouse model of rheumatoid arthritis

[0355] Effect of SP-C01 on acute paw edema in mice

[0356] As shown in Table 11, acute paw edema in mice was significantly increased in the model control group at 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 7d after administration (P≤0.01) compared with the normal group. In the SP-C01 low-dose group, paw edema was significantly reduced at 4h and 6h after administration (P≤0.01). In the SP-C01 medium-dose group, paw edema was significantly reduced at 2h, 4h, and 7d after administration (P≤0.01), and significantly reduced at 6h and 24h after administration (P≤0.05). In the SP-C01 high-dose group, paw edema was significantly reduced at 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 7d after administration (P≤0.01).

[0357] Effect of SP-C01 on the rate of increase in acute pain threshold in mice

[0358] As shown in Table 12, the acute pain threshold elevation rate in mice was significantly lower in the model control group than in the normal group at 0.5h, 1h, 2h, and 4h after administration (P≤0.01). Compared with the model control group, the pain threshold elevation rate in the low-dose SP-C01 group was significantly higher at 2h and 4h after administration (P≤0.01), and significantly higher at 1h after administration (P≤0.05). The pain threshold elevation rate in the medium-dose SP-C01 group was significantly higher at 1h, 2h, and 4h after administration (P≤0.01), and significantly higher at 0.5h after administration (P≤0.05). The pain threshold elevation rate in the high-dose SP-C01 group was significantly higher at 0.5h, 1h, 2h, and 4h after administration (P≤0.01).

[0359] Effect of SP-C01 on Pain Threshold in Mice

[0360] As shown in Table 13, the pain thresholds in mice with acute inflammation showed that, compared with the normal group, the pain thresholds in the model control group were significantly decreased at 0.5h, 1h, 2h, and 4h after administration (P≤0.01). Compared with the model control group, the pain thresholds in the low-dose SP-C01 group were significantly increased at 0.5h, 1h, 2h, and 4h after administration (P≤0.01); the pain thresholds in the medium-dose SP-C01 group were significantly increased at 0.5h, 1h, 2h, and 4h after administration (P≤0.01); and the pain thresholds in the high-dose SP-C01 group were significantly increased at 0.5h, 1h, 2h, and 4h after administration (P≤0.01). For mice with chronic inflammation, the pain thresholds in the model control group were significantly decreased compared with the normal group (P≤0.01), while the pain thresholds in the high-dose SP-C01 group were significantly increased compared with the model control group (P≤0.05).

[0361] Table 11 Effect of SP-C01 on acute paw edema in mice (n=10)

[0362]

[0363]

[0364] Note: Compared with the normal control group, ++P≤0.01; compared with the model control group, **P≤0.01, *P≤0.05.

[0365] Table 12 Effect of SP-C01 on the rate of increase in acute pain threshold in mice (n=10)

[0366]

[0367] Note: Compared with the normal control group, ++P≤0.01; compared with the model control group, **P≤0.01, *P≤0.05.

[0368] Table 13 Effect of SP-C01 on pain threshold in mice (n=10)

[0369]

[0370] Note: Compared with the normal control group, ++P≤0.01; compared with the model control group, **P≤0.01, *P≤0.05.

[0371] Effects of SP-C01 on Joint Index Scores in Mice with Chronic Inflammation

[0372] As shown in Table 14, the joint index scores of mice with chronic inflammation were significantly lower in the high-dose SP-C01 group on day 3 compared with the model control group (P≤0.05). On day 7, the joint index scores of mice with chronic inflammation were significantly lower in the low-dose SP-C01 group, medium-dose SP-C01 group, and high-dose SP-C01 group (P≤0.01).

[0373] Table 14 Effects of SP-C01 mice with chronic inflammation on joint index scores (n=10)

[0374]

[0375] Note: Compared with the model control group, **P≤0.01, *P≤0.05

[0376] Effects of SP-C01 on a rat model of chronic granulomatosis

[0377] Effects of SP-C01 on chronic granuloma in rats

[0378] As shown in Table 15, compared with the model control group, the chronic granulation swelling of rats in the low-dose SP-C01 group, medium-dose SP-C01 group, and high-dose SP-C01 group was significantly reduced (P≤0.01).

[0379] Table 15 Effects of SP-C01 on chronic granulomatosis in rats (n=10)

[0380]

[0381] Note: Compared with the normal control group, ++P≤0.01.

[0382] Experiment Summary

[0383] Effects of SP-C01 on a Fluorotrachet-induced rheumatoid arthritis model in mice: Under acute inflammatory conditions: In the low-dose SP-C01 group, paw swelling was significantly reduced at 4 and 6 hours after administration (P≤0.01), indicating that the low-dose SP-C01 had an antagonistic effect on inflammation in the Fluorotrachet-induced rheumatoid arthritis model mice at 4 and 6 hours after administration; The pain threshold of the low-dose SP-C01 was significantly increased at 0.5, 1, 2, and 4 hours after administration (P≤0.01), indicating that the low-dose SP-C01 had a significant analgesic effect on the Fluorotrachet-induced rheumatoid arthritis model mice at 0.5, 1, 2, and 4 hours after administration. At 2h, 4h, and 7d after administration of medium-dose SP-C01, mouse paw swelling was significantly reduced (P≤0.01), and at 6h and 24h after administration (P≤0.05), indicating that medium-dose SP-C01 had an antagonistic effect on inflammation in a mouse model of rheumatoid arthritis induced by Fluoride's complete adjuvant at 2h, 4h, 6h, 24h, and 7d after administration. At 0.5h, 1h, 2h, and 4h after administration of medium-dose SP-C01, the pain threshold in mice was significantly increased (P≤0.01), indicating that medium-dose SP-C01 had a significant analgesic effect in a mouse model of rheumatoid arthritis induced by Fluoride's complete adjuvant at 0.5h, 1h, 2h, and 4h after administration. The high-dose SP-C01 group showed a significant reduction in paw swelling in mice at 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 7d after administration (P≤0.01), indicating that the high-dose SP-C01 had an antagonistic effect on inflammation in a mouse model of rheumatoid arthritis induced by Fluoride's complete adjuvant at 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 7d after administration. The high-dose SP-C01 group also showed a significant increase in the rate of pain threshold elevation in mice at 0.5h, 1h, 2h, and 4h after administration (P≤0.01), indicating that the high-dose SP-C01 had a significant analgesic effect on a mouse model of rheumatoid arthritis induced by Fluoride's complete adjuvant at 0.5h, 1h, 2h, and 4h after administration.

[0384] In chronic inflammatory states: High-dose SP-C01 administration to mice for 7 consecutive days significantly increased the pain threshold (P≤0.05), indicating that high-dose SP-C01 administration for 7 consecutive days had a significant analgesic effect on fluoroadjuvant-induced rheumatoid arthritis model mice. On day 3 of administration, the joint index score of the high-dose SP-C01 group was significantly reduced (P≤0.05). After 7 days of administration, the joint index scores of the low-dose, medium-dose, and high-dose SP-C01 groups were all significantly reduced (P≤0.01), indicating that low-dose SP-C01 administration for 7 days, medium-dose SP-C01 administration for 7 days, and high-dose SP-C01 administration for 3 days and 7 days of administration had a regulatory effect on the paw joints of fluoroadjuvant-induced rheumatoid arthritis model mice.

[0385] Effects of SP-C01 on chronic granuloma in rats: Low-dose, medium-dose, and high-dose SP-C01 significantly reduced the degree of chronic granuloma swelling in rats (P≤0.01), indicating that low-dose, medium-dose, and high-dose SP-C01 have an effect on granuloma swelling in rats with chronic granuloma model.

[0386] The above results indicate that:

[0387] In acute inflammatory states, low-dose SP-C01 at 4h and 6h, medium-dose SP-C01 at 2h, 4h, 6h, 24h, and 7d, and high-dose SP-C01 at 1h, 2h, 4h, 6h, 8h, 12h, 24h, and 7d showed antagonistic effects on inflammation in a flurryl adjuvant-induced rheumatoid arthritis model mouse. In acute inflammatory states, low-dose SP-C01 at 0.5h, 1h, 2h, and 4h, and medium-dose SP-C01 at 0.5h, 1h, 2h, and 4h, and high-dose SP-C01 at 0.5h, 1h, 2h, and 7d also showed antagonistic effects on inflammation. SP-C01 high-dose administration (0.5h, 1h, 2h, 4h) and continuous administration for 7 days in mice under chronic inflammatory conditions significantly analgesic effects in a flurryl adjuvant-induced rheumatoid arthritis model. In contrast, the positive control drug celecoxib only showed analgesic effects 2 hours after administration in an acute inflammatory state. SP-C01's anti-inflammatory and analgesic effects in flurryl adjuvant-induced rheumatoid arthritis model mice had a significantly faster onset time and longer duration of action than celecoxib.

[0388] In a chronic inflammatory state, SP-C01 low-dose administration for 7 days, SP-C01 medium-dose administration for 7 days, SP-C01 high-dose administration for 3 days, and SP-C01 administration for 7 days all had a regulatory effect on the paw joints of a flurryl adjuvant-induced rheumatoid arthritis model mouse. Compared with the positive control drug celecoxib, the effects of all SP-C01 dose groups were stronger than those of celecoxib.

[0389] Low-dose, medium-dose, and high-dose SP-C01 inhibited granuloma swelling in a rat model of chronic granulomatosis. Compared with the positive control drug celecoxib, the low-dose and medium-dose SP-C01 groups showed comparable efficacy, while the high-dose SP-C01 group was more effective than celecoxib.

[0390] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. Other embodiments can be obtained based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. The use of a compound and its pharmaceutically acceptable salt in the preparation of a peroxisome proliferator-activated receptor agonist and a soluble epoxide hydrolase inhibitor, characterized in that, The compound has any one of the following structures: 、 、 、 、 。 2. Use according to claim 1, characterized in that, The peroxisome proliferator-activated receptor agonist and soluble epoxide hydrolase inhibitor are used for treating a soluble epoxide hydrolase and peroxisome proliferator-activated receptor mediated disease; The soluble epoxide hydrolase and peroxisome proliferator-activated receptor mediated disease is selected from an inflammatory disease, pain, sepsis, a cardiovascular disease, a neurodegenerative disease, diabetes, a diabetic complication, depression, liver fibrosis, renal failure, chronic obstructive pulmonary disease, or a pulmonary hypertension disease.

3. Use according to claim 2, characterized in that, The inflammatory disease includes non-alcoholic steatohepatitis or chronic nephritis; the pain includes neuropathic pain.

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