A multi-TRP ion channel modulator, its preparation method and application

Abstract

This application relates to the pharmaceutical field, and specifically to a multi-TRP ion channel modulator, its preparation method, and its application. The multi-TRP ion channel modulator developed in this application can inhibit TRPV1, activate TRPM8, and activate TRPA1, and can be used to treat functional gastrointestinal disorders, intestinal mucosal damage, intestinal inflammation, immune dysregulation, gut microbiota imbalance, gastrointestinal tumors, pain, or peripheral neuritis.

Description

Technical Field

[0001] This application relates to the pharmaceutical field, and in particular to a multi-TRP ion channel modulator, its preparation method, and its application. Background Technology

[0002] Inflammatory bowel disease (IBD) is a relapsing, chronic inflammatory bowel disease of unknown etiology, including ulcerative colitis (UC) and Crohn's disease (CD). UC primarily affects the colon and rectum, with lesions limited to the mucosal and submucosal layers; CD has a skip-onset pattern throughout the digestive tract, with lesions potentially affecting all layers. The pathogenesis of IBD may be related to genetic factors, environmental factors, and mucosal immune dysregulation.

[0003] Transient receptor potential (TRP) channels are classified into 6 groups and 26 subtypes: TRPA1, TRPV (1-6), TRPM (1-8), TRPML (1-3), TRPC (1, 3-7), and TRPP (2, 3, 5). TRP channels are ligand-gated, calcium-permeable, non-selective cation channels and are important temperature receptors. TRPV1, also known as the capsaicin receptor and vanillic acid receptor, can be activated by various physical and chemical stimuli, such as heat (temperature >43°C) and protons (pH <5.9). TRPM8 is a cold receptor, activated by low temperatures (<28°C), pressure, and cold-sensing compounds. TRPA1, also known as ANKTM1, is also a cold receptor and can be activated by many aromatic substances. Currently, TRP channels are extensively studied in neurological diseases and are considered important drug targets for pain and peripheral neuritis.

[0004] Current treatments primarily focus on anti-inflammatory and symptomatic relief, with no effective cure for the disease. Therefore, there is an urgent need to conduct scientific research to uncover the underlying pathogenesis of IBD in order to identify new therapeutic targets. Summary of the Invention

[0005] To address the aforementioned technical problems, this application presents a multi-TRP ion channel modulator that can inhibit TRPV1, activate TRPM8, and activate TRPA1. Its molecular structure is as follows (molecular weight: 827.95):

[0006] .

[0007] The applicant's previous research found that STW5, a traditional European herbal preparation used to treat functional gastrointestinal disorders, can regulate the activation and desensitization of intestinal primary sensory neurons TRPA1 and TRPV1 (PMID: 32682914); and that Menthacarin, a traditional European herbal preparation used to treat functional gastrointestinal disorders, can induce calcium influx and desensitization in macrophages, intestinal primary sensory neurons, and colonic organoids (PMID: 33127519). Further research revealed that the cold-sensing receptor TRPM8 reduces the release of substance P from primary sensory neurons by regulating the binding of PKAca to GSK-3β, thereby inhibiting substance P-induced epithelial cell apoptosis, alleviating intestinal mucosal damage, and relieving intestinal inflammation. It was also clarified that TRPM8 agonists (menthol) and substance P receptor antagonists (aprepitant) are potential drug options for intestinal inflammation and mucosal damage (PMID: 38280896). Furthermore, it was found that alcohol exacerbates intestinal inflammation and induces macrophage M2b polarization through TRPV1-MAPK / NF-κB. The TRPV1 inhibitor capsaicin (CPZ) is a potential drug option for intestinal inflammation. CPZ also has a certain activating effect on another TRP channel, TRPA1, and has potential therapeutic effects on intestinal inflammation (PMID: 38810558). Another study found that the TRPV1 agonist alcohol can induce circulatory system microbial imbalance, leading to a significant increase in Gram-negative bacteria and pro-inflammatory bacterial endotoxins in the body, damaging the colonic epithelial barrier function and aggravating the occurrence and development of esophageal cancer (PMID: 40001134).

[0008] In summary, TRP ion channels, especially TRPV1, TRPM8, and TRPA1, have certain therapeutic effects on functional gastrointestinal disorders, intestinal mucosal damage, intestinal inflammation, immune dysregulation, gut microbiota imbalance, gastrointestinal tumors, pain, and peripheral neuritis.

[0009] This application provides a compound or a pharmaceutically acceptable salt thereof, the chemical structural formula of which is shown in Formula XII:

[0010] Formula XII.

[0011] This application also provides a pharmaceutical composition comprising the above-described compound or a pharmaceutically acceptable salt thereof.

[0012] This application also provides the use of the above-described compounds or pharmaceutically acceptable salts or pharmaceutical compositions thereof in the preparation of medicaments.

[0013] This application also provides a method for preparing the above-mentioned compound, comprising the following steps:

[0014] 1) The compound shown in Formula I is mixed with HBr and reacted to obtain the compound shown in Formula II;

[0015] ;

[0016] 2) Mix the compound shown in Formula III and the compound shown in Formula IV, and react them to obtain the compound shown in Formula V;

[0017] ;

[0018] 3) The compound shown in formula V was mixed with TsCl, reacted, and extracted to obtain the compound shown in formula VI;

[0019] ;

[0020] 4) Mix the compound shown in Formula VI and the compound shown in Formula VII, and react them to obtain the compound shown in Formula VIII;

[0021] ;

[0022] 5) Dissolve the compound represented by formula VIII in a solvent and react to obtain the compound represented by formula IX;

[0023] ;

[0024] 6) Mix the compound shown in Formula IX and the compound shown in Formula X, and react them to obtain the compound shown in Formula XI;

[0025] ;

[0026] 7) Mix the compound shown in Formula XI and the compound shown in Formula II, and react them to obtain the compound shown in Formula XII;

[0027] .

[0028] The beneficial effects of the multi-TRP ion channel modulators proposed in this specification include, but are not limited to: (1) superior binding activity to TRPV1 compared to the specific inhibitor CPZ, superior binding activity to TRPM8 compared to the agonist Icilin, and superior binding activity to TRPA1 compared to the agonist Allyl isothiocyanate; (2) a regulatory effect on calcium ion influx in cells, and an effect similar to that of a mixture of TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, and TRPA1 agonist AITC; (3) no significant effect on normal macrophages, but can inhibit macrophage activation under inflammatory conditions; compared with individual TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, TRPA1 agonist AITC and their combinations, the multi-TRP ion channel modulators have a smaller effect on normal cells, but their inhibitory effect on macrophage activation under inflammatory conditions is similar to the above three; (4) a regulatory effect on inflammatory conditions. It inhibits the release of major pro-inflammatory factors in lower macrophages and promotes the release of anti-inflammatory factors. Its effect is consistent with the individual effects of TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, and TRPA1 agonist AITC, but it is better than the combined use of the three; (5) Its therapeutic effect on intestinal inflammation and mucosal damage in mice is better than the combined use of TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, and TRPA1 agonist AITC, and the effect of enema of multi-TRP ion channel regulator is better than that of oral administration; (6) It has little effect on the biochemistry, liver function, kidney function, blood lipids, etc. of mice, and significantly reduces the level of lactate dehydrogenase. Attached Figure Description

[0029] This application will be further described by way of exemplary embodiments, which will be described in detail with reference to the accompanying drawings. These embodiments are not limiting, wherein:

[0030] Figure 1 The binding pattern of Human TRPV1 protein with Compound1 is shown below. A. Cartoon image of Human TRPV1 protein binding with Compound1; B. Surface image of Human TRPV1 protein binding with Compound1; C. 2D image of Human TRPV1 protein binding with Compound1; D. 3D image of Human TRPV1 protein binding with Compound1.

[0031] Figure 2The binding patterns of Human TRPV1 protein and CPZ are illustrated below. A. Cartoon image of Human TRPV1 protein binding with CPZ; B. Surface image of Human TRPV1 protein binding with CPZ; C. 2D image of Human TRPV1 protein binding with CPZ; D. 3D image of Human TRPV1 protein binding with CPZ.

[0032] Figure 3 The binding pattern of Human TRPM8 protein with Compound1 is shown below. A. Cartoon image of Human TRPM8 protein binding with Compound1; B. Surface image of Human TRPM8 protein binding with Compound1; C. 2D image of Human TRPM8 protein binding with Compound1; D. 3D image of Human TRPM8 protein binding with Compound1.

[0033] Figure 4 The binding patterns of Human TRPM8 protein and Icilin are shown in the following diagrams: A. Cartoon image of Human TRPM8 protein binding with Icilin; B. Surface image of Human TRPM8 protein binding with Icilin; C. 2D image of Human TRPM8 protein binding with Icilin; D. 3D image of Human TRPM8 protein binding with Icilin.

[0034] Figure 5 A. Images showing changes in calcium influx intensity in macrophages under different stimuli, with calcium influx intensity represented by fluorescence intensity; B. Bar charts showing calcium influx intensity in macrophages under different stimuli, n=5.

[0035] Figure 6 A. Phosphorylation and non-phosphorylation levels of NF-κB and P38 proteins in unactivated and activated (LPS 1 μg / mL) macrophages after 6 h of stimulation with different concentrations of multi-TRP ion channel modulators; B. Phosphorylation and non-phosphorylation levels of NF-κB, P38, and Erk1 / 2 proteins in unactivated and activated (LPS 1 μg / mL) macrophages after 6 h of stimulation with 1 μM multi-TRP ion channel modulator, CPZ, AITC, Icilin, and a mixture of the three.

[0036] Figure 7The release of IL-1β, TNF-α, IL-23, IFN-γ, IL-12, and IL-10 from macrophages activated with LPS 1 μg / mL for 6 h after stimulation with compound 1 μM of multi-TRP ion channel modulator (Compound1), CPZ 1 μM, Icilin 1 μM, AITC 1 μM, and a mixture of CPZ, Icilin, and AITC (1 μM each). n=5.

[0037] Figure 8 A. Percentage weight curve changes of mice in different groups, n=5; B. Difference in percentage weight of mice in different groups on day 8, n=5; C. Comparison of colon length of mice in different groups, n=5; D. Rectal H&E staining images of mice in different groups; E. Colon images of mice in different groups.

[0038] Figure 9 A. H&E staining images of the liver and kidneys of the control group and mice after oral administration of different concentrations of multi-TRP ion channel modulators for 8 days; B. Serum biochemistry, liver function, kidney function, blood lipids and other indicators of mice in different groups, n=5; C. Bar chart comparing the differences of the above indicators, n=5.

[0039] Figure 10 The binding patterns of Human TRPA1 protein and Compound1 are shown in the following diagrams: A. Cartoon diagram of Human TRPA1 protein binding with Compound1; B. Surface diagram of Human TRPA1 protein binding with Compound1; C. 2D diagram of Human TRPA1 protein binding with Compound1; D. 3D diagram of Human TRPA1 protein binding with Compound1.

[0040] Figure 11 The binding patterns of Human TRPA1 protein with Allyl isothiocyanate are illustrated in the following diagrams: A. Cartoon image of Human TRPA1 protein binding with Allyl isothiocyanate; B. Surface image of Human TRPA1 protein binding with Allyl isothiocyanate; C. 2D image of Human TRPA1 protein binding with Allyl isothiocyanate; D. 3D image of Human TRPA1 protein binding with Allyl isothiocyanate. Detailed Implementation

[0041] To more clearly illustrate the technical solutions of the embodiments in this specification, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are merely some examples or embodiments of this specification. For those skilled in the art, these drawings can be applied to other similar scenarios without creative effort. Unless obvious from the context or otherwise specified, the same reference numerals in the drawings represent the same structures or operations.

[0042] As indicated in this specification and claims, unless the context clearly indicates otherwise, the words "a," "an," "an," and / or "the" do not specifically refer to the singular and may also include the plural. Generally speaking, the terms "comprising" and "including" only indicate the inclusion of expressly identified steps and elements, which do not constitute an exclusive list, and the method or apparatus may also include other steps or elements.

[0043] Flowcharts are used in this specification to illustrate the operations performed by the system according to embodiments of this specification. It should be understood that the preceding or following operations are not necessarily performed in exact order. Instead, the steps can be processed in reverse order or simultaneously. Furthermore, other operations can be added to these processes, or one or more steps can be removed from them.

[0044] This application provides a compound or a pharmaceutically acceptable salt thereof, the chemical structural formula of which is shown in Formula XII:

[0045] Formula XII.

[0046] This application also provides a pharmaceutical composition comprising the above-described compound or a pharmaceutically acceptable salt thereof.

[0047] In some embodiments, the pharmaceutical composition may further include a pharmaceutically acceptable carrier or excipient.

[0048] The term “pharmaceutically acceptable” as used in this article generally means a compound, material, composition, and / or dosage form that is suitable, within reasonable medical judgment, for contact with human and animal tissues, organs, and / or body fluids without excessive toxicity, irritation, allergic response, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0049] The excipients include various excipients and diluents, which are not essential active ingredients and do not cause excessive toxicity after application. The excipients contain sterile water or physiological saline, stabilizers, excipients, antioxidants (ascorbic acid, etc.), buffers (phosphate, citric acid, other organic acids, etc.), preservatives, surfactants (PEG, Tween, etc.), chelating agents (EDTA, etc.), or binders. The excipients also contain other low molecular weight peptides, serum albumin, glycine, glutamine, asparagine, arginine, polysaccharides, monosaccharides, mannitol, or sorbitol. When used in an aqueous solution for injection, the excipients are selected from physiological saline, isotonic glucose solution, D-sorbitol isotonic solution, D-mannose isotonic solution, D-mannitol or sugar alcohol isotonic solution. The aqueous solution for injection contains a solubilizer. The solubilizer is selected from alcohols (ethanol), polyols (propylene glycol or PEG), and / or nonionic surfactants (Tween 80 or HCO-50).

[0050] In the pharmaceutical composition provided in this application, the aforementioned compound may be a single active ingredient or may be combined with one or more other active components useful for the treatment of diseases to form a combined formulation.

[0051] The content of the active ingredient in the pharmaceutical composition is a safe and effective amount, which should be adjustable by those skilled in the art. For example, the dosage of the active ingredient in the above-mentioned compound and pharmaceutical composition depends on the patient's weight, the type of application, the condition and severity of the disease.

[0052] This application also provides the use of the above-described compounds or pharmaceutically acceptable salts or pharmaceutical compositions thereof in the preparation of medicaments.

[0053] In some embodiments, the drug may be a multi-TRP ion channel modulator.

[0054] In some embodiments, the drug can inhibit TRPV1, activate TRPM8, and activate TRPA1.

[0055] In some embodiments, the drug may be used to treat at least one of the following conditions: functional gastrointestinal disorders, intestinal mucosal damage, intestinal inflammation, immune dysregulation, gut microbiota imbalance, gastrointestinal tumors, pain, or peripheral neuritis.

[0056] In some embodiments, the drug may have one or more of the following functions:

[0057] Combined with TRPV1;

[0058] Regulates the influx of calcium ions into cells;

[0059] Inhibit macrophage activation under inflammatory conditions;

[0060] Inhibit the release of pro-inflammatory factors from macrophages in an inflammatory state;

[0061] Promotes the release of anti-inflammatory factors from macrophages in an inflammatory state;

[0062] Lower lactate dehydrogenase levels.

[0063] This application also provides a method for preparing the above-mentioned compound, comprising the following steps:

[0064] 1) The compound shown in Formula I is mixed with HBr and reacted to obtain the compound shown in Formula II;

[0065] ;

[0066] 2) Mix the compound shown in Formula III and the compound shown in Formula IV, and react them to obtain the compound shown in Formula V;

[0067] ;

[0068] 3) The compound shown in formula V was mixed with TsCl, reacted, and extracted to obtain the compound shown in formula VI;

[0069] ;

[0070] 4) Mix the compound shown in Formula VI and the compound shown in Formula VII, and react them to obtain the compound shown in Formula VIII;

[0071] ;

[0072] 5) Dissolve the compound represented by formula VIII in a solvent and react to obtain the compound represented by formula IX;

[0073] ;

[0074] 6) Mix the compound shown in Formula IX and the compound shown in Formula X, and react them to obtain the compound shown in Formula XI;

[0075] ;

[0076] 7) Mix the compound shown in Formula XI and the compound shown in Formula II, and react them to obtain the compound shown in Formula XII;

[0077] .

[0078] In some embodiments, the reaction conditions of step 1) can be 100–150 °C. o The reaction is carried out at C for 3–7 hours. In some embodiments, the reaction conditions of step 1) can be 105–145 °C.o The reaction is carried out at C for 3.5–6.5 h. In some embodiments, the reaction conditions of step 1) can be 110–140 °C. o The reaction is carried out at C for 4.0–6 hours. In some embodiments, the reaction conditions of step 1) can be 115–135 °C. o The reaction is carried out at C for 4.5–5.5 h. In some embodiments, the reaction conditions of step 1) can be 120–130 °C. o The reaction is carried out at C for 4.5–5 hours. In some embodiments, the reaction conditions of step 1) can be 125–130 °C. o The reaction proceeds at C for 4.5–5 hours.

[0079] In some embodiments, preferably, the reaction conditions of step 1) can be 130. o React at C for 5 hours.

[0080] In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (50~150) mg: (5~15) mL. In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (60~140) mg: (6~14) mL. In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (70~130) mg: (7~13) mL. In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (80~120) mg: (8~12) mL. In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (90~110) mg: (9~11) mL. In some embodiments, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be (100~110) mg: (10~11) mL.

[0081] In some embodiments, preferably, the mass-to-volume ratio of the compound represented by Formula I in step 1) to HBr can be 100 mg: 10 mL.

[0082] In some embodiments, the reaction conditions of step 2) can be heating under reflux in a closed device for 10 to 14 hours. For example, the reaction conditions of step 2) can be heating under reflux in a closed device for 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, or 14 hours. Any range characterized by combinations of the above values ​​is also included, which will not be elaborated here. In some embodiments, preferably, the reaction conditions of step 2) can be heating under reflux in a closed device for 12 hours.

[0083] In some embodiments, the molar ratio of the compound represented by Formula III and the compound represented by Formula IV in step 2) can be (0.5~1.5):(0.5~1.5).

[0084] In some embodiments, step 2) may further include a solvent and a carbonate.

[0085] In some embodiments, the solvent in step 2) can be acetonitrile, and the carbonate can be sodium carbonate.

[0086] In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (0.5~1.5):(0.5~1.5). In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (0.6~1.4):(0.6~1.4). In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (0.7~1.3):(0.7~1.3). In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (0.8~1.2):(0.8~1.2). In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (0.9~1.1):(0.9~1.1). In some embodiments, the molar ratio of the compound represented by formula V in step 3) to TsCl can be (1.0~1.1):(1.0~1.1).

[0087] In some embodiments, preferably, the molar ratio of the compound represented by formula V in step 3) to TsCl can be 1:1.

[0088] In some embodiments, step 3) may further include a solvent and triethylamine.

[0089] In some embodiments, the solvent in step 3) may be dichloromethane.

[0090] In some embodiments, the reaction conditions for step 3) can be stirring at room temperature for 12 hours.

[0091] In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (0.5~1.5):(0.5~1.5).

[0092] In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (0.6~1.4):(0.6~1.4). In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (0.7~1.3):(0.7~1.3). In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (0.8~1.2):(0.8~1.2). In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (0.9~1.1):(0.9~1.1). In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be (1.0~1.1):(1.0~1.1).

[0093] In some embodiments, the molar ratio of the compound represented by formula VI to the compound represented by formula VII in step 4) can be 1:1.

[0094] In some embodiments, step 4) may also include a solvent and a carbonate.

[0095] In some embodiments, the reaction conditions for step 4) can be heating under reflux for 10 to 14 hours in a closed device.

[0096] For example, the reaction conditions in step 4) can be heating under reflux in a closed device for 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, or 14 hours. It also includes any range characterized by combinations of the above values, which will not be elaborated here.

[0097] In some embodiments, the reaction conditions for step 4) can be heating under reflux for 12 hours in a closed device.

[0098] In some embodiments, the solvent in step 4) can be acetonitrile, and the carbonate can be sodium carbonate.

[0099] In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (150~300) mg: (5~15) mL. In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (170~280) mg: (6~14) mL. In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (190~260) mg: (7~13) mL. In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (200~250) mg: (8~12) mL. In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (220~230) mg: (9~11) mL. In some embodiments, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be (220~230) mg: (10~11) mL.

[0100] In some embodiments, preferably, in step 5), the mass-to-volume ratio of the compound represented by formula VIII to the solvent can be 220 mg: 10 mL.

[0101] In some embodiments, the reaction conditions for step 5) can be room temperature for 0.5 to 2 hours. For example, the reaction conditions for step 5) can be room temperature for 0.5, 0.75, 1, 1.25, 1.5, 1.75, or 2 hours. Any range characterized by combinations of the above values ​​is also included, which will not be elaborated here.

[0102] In some embodiments, preferably, the reaction conditions for step 5) can be room temperature for 1 hour.

[0103] In some embodiments, in step 5), the solvent may be trifluoroacetic acid or dichloromethane.

[0104] In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (0.5~1.5):(0.5~1.5). In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (0.6~1.4):(0.6~1.4). In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (0.7~1.3):(0.7~1.3). In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (0.8~1.2):(0.8~1.2). In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (0.9~1.1):(0.9~1.1). In some embodiments, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be (1.0~1.1):(1.0~1.1).

[0105] In some embodiments, preferably, in step 6), the molar ratio of the compound represented by formula IX to the compound represented by formula X can be 1:1.

[0106] In some embodiments, step 6) may further include a solvent and triethylamine. In some embodiments, the solvent in step 6) may be dichloromethane.

[0107] In some embodiments, the reaction conditions for step 6) can be a reaction at room temperature for 0.5 to 2 hours. For example, the reaction conditions for step 6) can be a reaction at room temperature for 0.5, 0.75, 1, 1.25, 1.5, 1.75, or 2 hours. Any range characterized by combinations of the above values ​​is also included, which will not be elaborated here.

[0108] In some embodiments, preferably, the reaction conditions for step 6) can be a reaction at room temperature for 1 hour.

[0109] In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (0.5~1.5):(0.5~1.5). In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (0.6~1.4):(0.6~1.4). In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (0.7~1.3):(0.7~1.3). In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (0.8~1.2):(0.8~1.2). In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (0.9~1.1):(0.9~1.1). In some embodiments, the molar ratio of the compound represented by Formula XI to the compound represented by Formula II in step 7) can be (1.0~1.1):(1.0~1.1).

[0110] In some embodiments, preferably, the molar ratio of the compound represented by formula XI to the compound represented by formula II in step 7) can be 1:1.

[0111] In some embodiments, step 7) may further include a solvent and triethylamine. In some embodiments, the solvent in step 7) may be dichloromethane.

[0112] In some embodiments, the reaction conditions for step 7) can be a reaction at room temperature for 0.5 to 2 hours. For example, the reaction conditions for step 7) can be a reaction at room temperature for 0.5, 0.75, 1, 1.25, 1.5, 1.75, or 2 hours. Any range characterized by combinations of the above values ​​is also included, which will not be elaborated here.

[0113] In some embodiments, preferably, the reaction conditions for step 7) can be a reaction at room temperature for 1 hour.

[0114] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were all purchased from conventional biochemical reagent companies. All quantitative experiments in the following examples were performed in triplicate or at least, and the results were averaged.

[0115] Example 1 - Preparation method is as follows:

[0116]

[0117] 1) Dissolve HM-5036_1 (100 mg, 0.41 mmol, 1.0 eq) in 48% HBr (10 mL), 130 oThe reaction was carried out at C for 5 hours, and the reaction progress was monitored by LCMS. After the reactants had basically reacted, the reaction apparatus was dismantled, and the product was concentrated to obtain a pale yellow solid, 109 mg of crude product.

[0118]

[0119] 2) HM-5036_3 (660 mg, 2.78 mmol, 1.0 eq), HM-417_15 (969 mg, 2.78 mmol, 1.0 eq), acetonitrile (30 mL), and K2CO3 (1537 mg, 11.1 mmol, 4 eq) were added to a 100 mL single-necked flask and refluxed for 12 h. The reaction progress was monitored by LCMS. After the reactants had basically reacted, the reaction apparatus was removed, the filtrate was filtered, and the crude product was concentrated. The crude product was purified by column chromatography (EA / MeOH (V / V=1 / 4)) to obtain 910 mg of a colorless oil.

[0120]

[0121] 3) HM-5036_4 (900 mg, 2.18 mmol, 1.0 eq), TsCl (415 mg, 2.18 mmol, 1.0 eq), DCM (20 mL), and TEA (440 mg, 4.36 mmol, 2 eq) were added to a 50 mL single-necked flask and stirred at room temperature for 12 h. The reaction progress was monitored by LCMS. After the reactants had basically reacted, the reaction apparatus was removed, and the reaction solution was poured into water (50 mL). DCM (20 mL x 3) was used for extraction. The combined organic phases were washed with saturated NaCl, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (EA / MeOH (V / V=1 / 6)) to obtain 588 mg of white solid.

[0122]

[0123] 4) Add HM-5036_5 (400 mg, 0.7 mmol, 1.0 eq), HM-5036_6 (220 mg, 0.7 mmol, 1.0 eq), acetonitrile (20 mL), and K2CO3 (440 mg, 2.8 mmol, 4.0 eq) to a 50 mL single-necked flask, reflux for 12 h, and monitor the reaction progress with LCMS. Once the reactants have largely reacted, dismantle the reaction apparatus, filter to obtain the filtrate, and concentrate to obtain the crude product. Purify by column chromatography (EA / MeOH (V / V=1 / 4)) to obtain a colorless oil, 240 mg, yield 48%.

[0124]

[0125] 5) HM-5036_7 (220 mg, 0.31 mmol, 1.0 eq) was dissolved in TFA / DCM (10 mL / 10 mL) and reacted at room temperature for 1 h. The reaction progress was monitored by LCMS. After the starting materials had basically reacted, the crude product was concentrated and purified by column chromatography (EA / MeOH (V / V=1 / 3)) to obtain 170 mg of crude product as a pale yellow oil.

[0126]

[0127] 6) Add HM-5036_8 (170 mg, 0.28 mmol, 1.0 eq), HM-5036_9 (65 mg, 0.28 mmol, 1.0 eq), DCM (20 mL), and TEA (57 mg, 0.56 mmol, 2.0 eq) to a 100 mL single-necked flask and react at room temperature for 1 h. Monitor the reaction progress using LCMS. Once the reactants have largely reacted, dismantle the reaction apparatus, filter to obtain the filtrate, and concentrate to obtain the crude product. Purify by column chromatography (EA / MeOH (V / V=1 / 4)) to obtain 100 mg of crude product as a yellow solid.

[0128]

[0129] 7) Add HM-5036_10 (100 mg, 0.15 mmol, 1 eq), HM-5036_2 (40 mg, 0.15 mmol, 1 eq), DCM (20 mL), and TEA (31 mg, 0.3 mmol, 2 eq) to a 100 mL single-necked flask. Incubate at room temperature for 1 h, monitoring the reaction progress with LCMS. Once the reactants have largely reacted, dismantle the reaction apparatus and concentrate at room temperature to obtain the crude product. Purify by Prep-HPLC and freeze-dry under reduced pressure to obtain 2 mg of white solid.

[0130]

[0131] Table 1

[0132]

[0133] Table 2

[0134]

[0135] Example 2 - The TRP channel modulation effect was detected as follows:

[0136] 1) Molecular docking:

[0137] process:

[0138] 1. Protein Preparation: The 3D structures of Human TRPV1, Human TRPM8, and Human TRPA1 were downloaded from the RCSB PDB database (Human TRPV1 PDB ID: 8JQR; Human TRPM8 PDB ID: 8BDC; Human TRPA1 PDB ID: 6PQO). The Protein Preparation Wizard module was used to hydrogenate the Human TRPV1, Human TRPM8, and Human TRPA1 proteins, respectively, removing other small molecules, protein chains, and ligand molecules, retaining only the A chain and its ligand. Energy optimization was then performed (OPLS2005 force field, RMSD 0.30 Å).

[0139] 2. Define the pocket grid file: Use the Receptor Grid Generation module to create a grid file for the processed protein. Generate a grid file centered on the binding pockets of Human TRPV1 antagonist ligand SAF312, Human TRPM8 small molecule ligand POV / Y01, and Human TRPA1 covalent agonist JT010. Set the box size to 20 Å × 20 Å × 20 Å.

[0140] 3. Ligand molecule preparation: The 2D structures of small molecule compounds such as multi-TRP ion channel modulators, TRPV1 inhibitors (CPZ), TRPM8 agonists (Icilin), and TRPA1 agonists (Allyl isothiocyanate) are processed by the LigPrep module for energy minimization (OPLS2005 force field, RMSD of 0.30 Å) to output the corresponding 3D structures.

[0141] 4. Molecular docking:

[0142] a. The high-precision (XP) mode of the Ligand docking module is used to enable molecular docking of the Human TRPV1 protein with the multi-TRP ion channel regulator and CPZ. That is, the multi-TRP ion channel regulator, CPZ and Human TRPV1 protein dock with each other through geometric matching and energy matching.

[0143] b. The high-precision (XP) mode of the Ligand docking module is used to enable molecular docking of the Human TRPM8 protein with the multi-TRP ion channel regulator and Icilin. That is, the multi-TRP ion channel regulator, Icilin and the Human TRPM8 protein dock with each other through geometric matching and energy matching.

[0144] c. The high-precision (XP) mode of the Ligand docking module is used to enable molecular docking of Human TRPA1 protein with multi-TRP ion channel regulators and Allyl isothiocyanate. That is, multi-TRP ion channel regulators, Allylisothiocyanate and Human TRPA1 protein dock with each other through geometric matching and energy matching.

[0145] result:

[0146] 1. Molecular docking results

[0147] The docking scores of Human TRPV1, Human TRPM8, and Human TRPA1 with small molecule compounds are shown in Table 3. (The docking score is an important indicator for evaluating the binding ability of proteins to small molecule compounds; the higher the absolute value, the stronger the binding force between the compound and the protein.)

[0148] Table 3

[0149] Protein Name Compound Docking Score Human TRPV1 (PDB ID: 8JQR) Compound1 -8.183 Human TRPV1 (PDB ID: 8JQR) HY-15640 -6.052 Human TRPM8 (PDB ID: 8BDC) Compound1 -8.760 Human TRPM8 (PDB ID: 8BDC) HY-11062 -6.579 Human TRPA1 (PDB ID: 6PQO) Compound1 -5.599 Human TRPA1 (PDB ID: 6PQO) HY-B1515 -1.474

[0150] Compound1: Multi-TRP ion channel modulator; HY-15640: CPZ; HY-11062: Icilin; HY-B1515: Allyl isothiocyanate

[0151] 2. Plotting the virtual screening results

[0152] a. Human TRPV1-multi-TRP ion channel modulator

[0153] 2D and 3D mapping was performed on the binding pattern of the Human TRPV1 multi-TRP ion channel regulator (docking score: -8.183) to the Human TRPV1 protein. The interaction pattern between the multi-TRP ion channel regulator and Human TRPV1 is shown in the figure below. Figure 1 As shown in A and B, the Human TRPV1 protein is displayed as a blue cartoon, and the multi-TRP ion channel regulator is displayed as a magenta stick. Figure 1In diagram D, the Human TRPV1 protein C backbone is shown in blue, N atoms in blue, O atoms in bright red, H atoms in white, multi-TRP ion channel regulators in magenta sticks, π-π interactions in green dashed lines, and hydrogen bonding interactions in purple dashed lines. The longer the hydrogen bond, the weaker the hydrogen bond interaction. Figure 1 As shown in C and D, the multi-TRP ion channel regulator can form one hydrogen bond and two π-π interactions with the Human TRPV1 protein: the phenolic hydroxyl group acts as a hydrogen bond donor to form one hydrogen bond with ASN551 at a distance of 1.9 Å; in addition, the multi-TRP ion channel regulator can also form two π-π interactions with TYR511.

[0154] b. Human TRPV1-CPZ

[0155] 2D and 3D mappings were performed to show the binding pattern of Human TRPV1-CPZ (docking score: -6.052) to Human TRPV1 protein. The interaction patterns between multi-TRP ion channel regulators and Human TRPV1 are shown below. Figure 2 As shown in A and B, the Human TRPV1 protein is displayed as a blue cartoon, and the multi-TRP ion channel regulator is displayed as a yellow stick. Figure 2 In diagram D, the Human TRPV1 protein C backbone is shown in blue, N atoms in blue, O atoms in bright red, H atoms in white, CPZ in yellow sticks, and π-π interactions in green dashed lines. (For example...) Figure 2 As shown in C and D, CPZ can form a π-π interaction with the Human TRPV1 protein: a benzene ring of the multi-TRP ion channel regulator can form a π-π interaction with TYR511.

[0156] c. Human TRPM8-multi-TRP ion channel modulator

[0157] 2D and 3D mapping was performed on the binding pattern of the human TRPM8 multi-TRP ion channel regulator (docking score: -8.760) to the human TRPM8 protein. The interaction pattern between the multi-TRP ion channel regulator and human TRPM8 is shown in the figure. Figure 3 As shown in A and B, the Human TRPM8 protein is displayed as a gray-white cartoon, and the multi-TRP ion channel regulator is displayed as a magenta stick. Figure 3In diagram D, the Human TRPM8 protein C backbone is shown as grayish-white, N atoms as blue, O atoms as bright red, H atoms as white, multi-TRP ion channel regulators as magenta sticks, π-π interactions as green dashed lines, and hydrogen bonding interactions as purple dashed lines. The longer the hydrogen bond length, the weaker the hydrogen bond interaction. Figure 3 As shown in C and D, the multi-TRP ion channel regulator can form one hydrogen bond and one π-π interaction with the Human TRPM8 protein: the phenolic hydroxyl group acts as a hydrogen bond donor to form one hydrogen bond with SER739 at a distance of 1.8 Å; in addition, the multi-TRP ion channel regulator can also form one π-π interaction with TRP740.

[0158] d. Human TRPM8-Icilin

[0159] 2D and 3D mappings were performed to illustrate the binding pattern between Human TRPM8-Icilin (docking score: -6.579) and Human TRPM8 protein. The interaction pattern between Icilin and Human TRPM8 is shown in the figure below. Figure 4 As shown in A and B, the Human TRPM8 protein is displayed as a gray-white cartoon, and the multi-TRP ion channel regulator is displayed as a green stick. Figure 4 In Figure D, the Human TRPM8 protein C backbone is shown as grayish-white, N atoms as blue, O atoms as bright red, H atoms as white, and Icilin as a green stick. π-π interactions are shown as green dashed lines, and hydrogen bonding interactions as purple dashed lines. The longer the hydrogen bond length, the weaker the hydrogen bond interaction. Figure 4 As shown in C and D, Icilin can form one hydrogen bond and one π-π interaction with the Human TRPM8 protein: the amino group on the six-membered ring acts as a hydrogen bond donor to form one hydrogen bond with SER739 at a distance of 2.1 Å; in addition, Icilin can also form one π-π interaction with PHE738.

[0160] e. Human TRPA1-Multi-TRP Ion Channel Modulator

[0161] 2D and 3D mapping was performed on the binding pattern of Human TRPA1-multi-TRP ion channel regulator (docking score: -5.599) to Human TRPA1 protein. The interaction pattern between the multi-TRP ion channel regulator and Human TRPA1 is shown in the figure. Figure 10As shown in Figures A and B, the Human TRPA1 protein C backbone is displayed in light blue, N atoms in blue, O atoms in bright red, H atoms in white, and the multi-TRP ion channel regulator is displayed as a light brown stick. Hydrogen bonding is shown as a yellow dashed line, and π-π conjugation is shown as a green dashed line; the longer the bond length, the weaker the hydrogen bond. Figure 10 As shown in C and D, the multi-TRP ion channel regulator can form two hydrogen bonds, two π-π conjugation interactions, and hydrophobic interactions with the Human TRPA1 protein: the hydroxyl group acts as a hydrogen bond donor, forming two hydrogen bonds with TYR680 and HIS614 at distances of 2.3 Å and 3.3 Å, respectively; the two benzene rings on the multi-TRP ion channel regulator form π-π conjugation interactions with TYR680 and PHE612. Furthermore, the multi-TRP ion channel regulator can also form hydrophobic interactions with surrounding amino acid residues.

[0162] f. Human TRPA1- Allyl isothiocyanate

[0163] 2D and 3D mapping was performed on the binding pattern of Human TRPA1-Allyl isothiocyanate (docking score: -1.474) with Human TRPA1 protein. The interaction pattern between Allyl isothiocyanate and Human TRPA1 is shown in the figure. Figure 11 As shown in Figures A and B, the Human TRPA1 protein C backbone is displayed in light blue, N atoms in blue, O atoms in bright red, H atoms in white, and Allyl isothiocyanate in yellow stick. Figure 11 As shown in C and D, Allyl isothiocyanate can form hydrophobic interactions with amino acid residues such as ILE627, CYS621, TYR662, and GLN664 of the Human TRPA1 protein.

[0164] The results showed that the multi-TRP ion channel modulators had a better binding effect on TRPV1 than the specific inhibitor CPZ, a better binding effect on TRPM8 than the agonist Icilin, and a better binding effect on TRPA1 than the agonist Allylisothiocyanate.

[0165] 2) Detection of calcium ion influx:

[0166] Procedure: RAW264.7 macrophage cells were used for detection. After stable passage culture, cells were cultured in serum-free medium for 1 hour and then loaded with Fluo-4 AM calcium influx reagent for 0.5 h. After removing the loading liquid, cells were cultured in serum-free medium for half an hour before the experiment. Cells were stimulated with serum-free medium containing different stimuli. The stimuli were changed after each stimulation. The fluorescence intensity at 488 nm wavelength of the cells under different stimuli was recorded by microscope, which is the calcium influx intensity. The stimuli and their concentrations were: Compound 1 μM of multi-TRP ion channel modulator, CPZ 1 μM, Icilin 1 μM, Allylisothiocyanate (AITC) 1 μM, and combinations thereof.

[0167] result: Figure 5 Image A shows images of fluorescence changes in the same macrophage under different stimuli. Figure 5 B. The fluorescence intensity change bar chart shows that multi-TRP ion channel modulators can increase cellular calcium influx, which is similar to the calcium influx intensity after CPZ + Icilin + AITC stimulation. Icilin + AITC stimulation further increases the calcium influx intensity, while CPZ + Icilin can decrease the cellular calcium influx intensity. CPZ + AITC can increase the cellular calcium influx intensity, but not as much as multi-TRP ion channel modulators. CPZ alone stimulates the lowest calcium influx intensity, and Icilin alone can increase the cellular calcium influx intensity, but not as much as AITC.

[0168] In summary, multi-TRP ion channel modulators regulate the influx of calcium ions into cells, and their effects are similar to those of a mixture of TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, and TRPA1 agonist AITC.

[0169] II. The following are the assay results for cell function regulation:

[0170] 3) Macrophage functional pathway activation status:

[0171] Procedure: RAW264.7 macrophage cells were used for detection. Stable passaged cells were cultured in serum-free medium for 1 hour, then stimulated for 6 hours with serum-free medium containing different stimulants. Cells were collected and Western blot was performed to detect phosphorylated and non-phosphorylated NF-κB, p38, and Erk1 / 2. The stimulants and their concentrations were: Compound 100 nM / 1μM / 10μM, LPS 1μg / mL, CPZ 1μM, Icilin 1μM, Allylisothiocyanate (AITC) 1μM, and combinations of CPZ, Icilin, and AITC.

[0172] Result: As Figure 6 As shown in Figure A, initial stimulation of cells with a multi-TRP ion channel modulator (100 nM / 1 μM / 10 μM) revealed no significant effect on the molecular levels and phosphorylation levels of NF-κB and p38 in normal cells. Further stimulation with LPS to simulate an inflammatory state showed that the multi-TRP ion channel modulator (100 nM / 1 μM / 10 μM) inhibited both NF-κB and p38 molecular levels, indicating its inhibitory effect on macrophage activation under inflammatory conditions. The effects of the multi-TRP ion channel modulator on CPZ, Icilin, and AITC were then compared using different stimuli. Figure 6 B found that CPZ significantly inhibited NF-κB phosphorylation in normal cells, affecting normal cell function. The combined use of CPZ, Icilin, and AITC, as well as AITC alone, significantly promoted p38 phosphorylation in normal cells, affecting normal cell function. The combined use of these three drugs, AITC alone, and Icilin alone significantly promoted NF-κB and Erk1 / 2 phosphorylation in normal cells, affecting normal cell function. Under LPS stimulation, the application of multi-TRP ion channel modulators, the combined use of the three molecules, and CPZ significantly inhibited NF-κB phosphorylation and suppressed macrophage activation under inflammatory conditions. Multi-TRP ion channel modulators and CPZ significantly inhibited p38 phosphorylation and suppressed macrophage activation under inflammatory conditions. Multi-TRP ion channel modulators, CPZ, and AITC significantly inhibited Erk1 / 2 phosphorylation and suppressed macrophage activation under inflammatory conditions.

[0173] In summary, multiple concentrations of multi-TRP ion channel modulators had no significant effect on normal macrophages, but they could inhibit macrophage activation under inflammatory conditions. Compared with individual TRPV1 inhibitors CPZ, TRPM8 agonists Icilin, TRPA1 agonists AITC, and their combinations, multi-TRP ion channel modulators had less effect on normal cells, but their inhibitory effect on macrophage activation under inflammatory conditions was similar to the above three.

[0174] 4) Macrophage inflammatory factor release status:

[0175] Procedure: Detection using macrophage cell lines RAW264.7Cells that had been stably passaged were cultured in serum-free medium for 1 hour, then stimulated for 6 hours with 1 μg / mL LPS (simulating an inflammatory state) and serum-free medium containing different stimulants. The supernatant was collected and ELISA was used to detect the concentrations of pro-inflammatory cytokines IL-1β, TNF-α, IL-23, IFN-γ, and IL-12, and the concentration of the anti-inflammatory cytokine IL-10. The stimulants and their concentrations were: Compound 1 μM (a multi-TRP ion channel modulator), CPZ 1 μM, Icilin 1 μM, Allyl isothiocyanate (AITC) 1 μM, and combinations of CPZ, Icilin, and AITC.

[0176] Result: As Figure 7 As shown, stimulation by multi-TRP ion channel modulators significantly inhibited the release of pro-inflammatory factors other than IL-12 from macrophages under inflammatory conditions, and significantly promoted the release of the anti-inflammatory factor IL-10. In contrast, CPZ mainly inhibited the release of IFN-γ, Icilin mainly inhibited the release of IL-1β and TNF-α, but promoted the release of IL-23, AITC mainly inhibited the release of IL-1β, TNF-α, and IFN-γ, and promoted the release of IL-10, while the combination of the three promoted the release of IL-23.

[0177] In summary, multi-TRP ion channel modulators inhibit the release of major pro-inflammatory factors in macrophages under inflammatory conditions and promote the release of anti-inflammatory factors. Their effects are consistent with those of the TRPV1 inhibitor CPZ, the TRPM8 agonist Icilin, and the TRPA1 agonist AITC, but they are more effective than the combined use of the three.

[0178] III. Animal experiments are as follows:

[0179] 1) Regulatory effect on intestinal inflammation:

[0180] Procedure: Eight-week-old C57BL6 mice were used to model the disease. They were given 2.5% DSS in drinking water for 7 days, and their body weight was measured daily during this period. On the 8th day, the mice were sacrificed, and serum, colon length was measured, and the last 0.5 cm of rectum was taken for H&E staining. The indicators were compared. The administration methods were as follows: 100 μL of PBS solution containing 10 μM, 100 μM, and 1 mM of the multi-TRP ion channel modulator (Compound1) was administered orally, equivalent to 41.3975 μg / kg / day, 413.975 μg / kg / day, and 4.13975 mg / kg / day, respectively; 100 μL of PBS solution containing a mixture of 10 μM, 100 μM, and 1 mM of CPZ, Icilin, and AITC was administered orally, equivalent to CPZ 18.845 μg / kg / day, 188.45 μg / kg / day, and 1.8845 mg / kg / day; Icilin 15.5645 μg / kg / day, 155.645 μg / kg / day, and 1.55645 mg / kg / day, respectively; AITC 4.9575 μg / kg / d, 49.575 μg / kg / d, 495.75 μg / kg / d; 100 μL of PBS solution containing 10 μM of multi-TRP ion channel modulator (Compound1) was administered via enema, which is equivalent to 41.3975 μg / kg / d.

[0181] Result: As Figure 8 As shown in the AE, the control group of the enteritis mouse model exhibited the most severe intestinal inflammation and mucosal damage, with the most significant colonic shortening. In the treatment groups, the 1mM multi-TRP ion channel modulator and the 1mM mixture of CPZ, Icilin, and AITC showed the best therapeutic effects, with the 100μM multi-TRP ion channel modulator showing similar efficacy. Furthermore, compared to the 100μM mixture of CPZ, Icilin, and AITC, it resulted in less mucosal damage. Figure 8 (BD). It is worth noting that an enema of 10 μM of multi-TRP ion channel modulators can achieve similar therapeutic effects as an oral administration of 1 mM of multi-TRP ion channel modulators. Figure 8 BD).

[0182] In summary, the therapeutic effect of multi-TRP ion channel modulators on intestinal inflammation and mucosal damage in mice is slightly better than the combined application of TRPV1 inhibitor CPZ, TRPM8 agonist Icilin, and TRPA1 agonist AITC, and the effect of multi-TRP ion channel modulators via enema is better than that via oral administration.

[0183] 2) Mouse toxicity test:

[0184] Procedure: After the mice in the experimental control group and the groups that were orally administered the multi-TRP ion channel modulator (Compound1) at 10 μM, 100 μM and 1 mM were sacrificed, their serum was used to measure biochemical, liver function, kidney function and blood lipid indicators. The liver and kidneys were stained with H&E to comprehensively evaluate drug toxicity.

[0185] Results: Liver and kidney staining results are as follows Figure 9 As shown in Figure A, there were no significant differences between the groups. Biochemical, liver function, kidney function, and blood lipid results are as follows: Figure 9 As shown in B, some inflammation and injury-related indicators were alleviated to some extent in the treatment group. For example... Figure 9 As shown in Figure C, the comparison revealed that the total protein and albumin levels were significantly improved in the 1 mM group. Except for the 1 mM group, the total bilirubin was slightly decreased, possibly due to anemia caused by enteritis bleeding. Alkaline phosphatase was slightly increased in the 100 μM and 1 mM groups, while lactate dehydrogenase was significantly higher in the control group than in the treatment group, reflecting the reduction of inflammation and tissue damage. The total bile acid was slightly increased in the 100 μM and 1 mM groups, which could also be explained by the aggravation of enteritis in other groups leading to anemia. Other indicators showed no significant differences.

[0186] In summary, multi-TRP ion channel modulators have minimal effects on biochemistry, liver function, kidney function, and blood lipids in mice, have a slight effect on bile acid and bilirubin metabolism, but can significantly reduce lactate dehydrogenase levels.

[0187] The basic concepts have been described above. Obviously, for those skilled in the art, the detailed disclosure above is merely illustrative and does not constitute a limitation of this specification. Although not explicitly stated herein, those skilled in the art may make various modifications, improvements, and corrections to this specification. Such modifications, improvements, and corrections are suggested in this specification and therefore remain within the spirit and scope of the exemplary embodiments described herein.

[0188] Furthermore, this specification uses specific terms to describe embodiments thereof. For example, "an embodiment," "one embodiment," and / or "some embodiments" refer to a particular feature, structure, or characteristic associated with at least one embodiment of this specification. Therefore, it should be emphasized and noted that references to "an embodiment," "one embodiment," or "an alternative embodiment" in different locations throughout this specification do not necessarily refer to the same embodiment. Moreover, certain features, structures, or characteristics in one or more embodiments of this specification can be appropriately combined.

[0189] In some embodiments, numbers describing the quantity of components and attributes are used. It should be understood that such numbers used in the description of embodiments are modified in some examples with the terms "approximately," "approximately," or "generally." Unless otherwise stated, "approximately," "approximately," or "generally" indicates that the numbers are allowed to vary by ±20%. Accordingly, in some embodiments, the numerical parameters used in the specification and claims are approximate values, which may be changed depending on the characteristics required by individual embodiments. In some embodiments, numerical parameters should take into account specified significant digits and employ a general method of digit reservation. Although the numerical ranges and parameters used to confirm their breadth of range in some embodiments of this specification are approximate values, in specific embodiments, such values ​​are set as precisely as feasible.

[0190] Finally, it should be understood that the embodiments described in this specification are merely illustrative of the principles of the embodiments described herein. Other variations may also fall within the scope of this specification. Therefore, alternative configurations of the embodiments described herein are intended to be illustrative rather than limiting, and should be considered consistent with the teachings of this specification. Accordingly, the embodiments described herein are not limited to those explicitly introduced and described herein.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, the chemical structure of the compound is shown as formula XX: Formula XI.

2. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to claim 1.

3. The pharmaceutical composition of claim 2, wherein Also comprising pharmaceutically acceptable adjuvants.

4. Use of the compound or a pharmaceutically acceptable salt thereof according to claim 1 or the pharmaceutical composition according to claim 2 or 3 in the preparation of a medicament for treating intestinal mucosal injury or intestinal inflammation.

5. The method for preparing the compound according to claim 1, characterized in that, Comprising the following steps: 1) mixing the compound shown as formula I with HBr, reacting to obtain the compound shown as formula II; ； 2) mixing the compound shown as formula III with the compound shown as formula IV, reacting to obtain the compound shown as formula V; ； 3) mixing the compound shown as formula V with TsCl, reacting, extracting to obtain the compound shown as formula VI; ； 4) mixing the compound shown as formula VI with the compound shown as formula VII, reacting to obtain the compound shown as formula VIII; ； 5) dissolving the compound shown as formula VIII in a solvent, reacting to obtain the compound shown as formula IX; ； 6) mixing the compound shown as formula IX with the compound shown as formula X, reacting to obtain the compound shown as formula XI; ； 7) mixing the compound shown as formula XI with the compound shown as formula II, reacting to obtain the compound shown as formula XII; 。 6. The production method according to claim 5, wherein The reaction conditions of the step 1) are 100-150 o C for 3-7 h; And / or, the mass-volume ratio of the compound shown as formula I to HBr in step 1) is (50-150) mg:(5-15) mL; And / or, the reaction condition of step 2) is heating and refluxing in a closed device for 10-14 h; And / or, the molar ratio of the compound shown as formula III to the compound shown as formula IV in step 2) is (0.5-1.5):(0.5-1.5); And / or, step 2) further comprises a solvent and a carbonate; And / or, the molar ratio of the compound shown as formula V to TsCl in step 3) is (0.5-1.5):(0.5-1.5); And / or, step 3) further comprises a solvent and triethylamine; And / or, the reaction condition of step 3) is stirring at room temperature for 12 h; And / or, the molar ratio of the compound shown as formula VI to the compound shown as formula VII in step 4) is (0.5-1.5):(0.5-1.5); And / or, step 4) further comprises a solvent and a carbonate; And / or, the reaction condition of step 4) is heating and refluxing in a closed device for 10-14 h; And / or, in step 5), the mass-volume ratio of the compound shown as formula VIII to the solvent is (150-300) mg:(5-15) mL; And / or, the reaction condition of step 5) is reacting at room temperature for 0.5-2 h; And / or, in step 6), the molar ratio of the compound shown as formula IX to the compound shown as formula X is (0.5-1.5):(0.5-1.5); And / or, step 6) further comprises a solvent and triethylamine; And / or, the reaction condition of step 6) is reacting at room temperature for 0.5-2 h; And / or, the molar ratio of the compound of formula IX to the compound of formula II in step 7) is (0.5-1.5):(0.5-1.5); And / or, step 7) further comprises a solvent and triethylamine; And / or, the reaction condition of step 7) is room temperature reaction for 0.5-2h.

7. The production method according to claim 6, wherein The reaction conditions of step 1) are 130 o C for 5 h; And / or, the reaction condition of step 2) is heating reflux for 12h in a closed device; And / or, the solvent in step 2) is acetonitrile, and the carbonate is sodium carbonate; And / or, the solvent in step 3) is dichloromethane; And / or, the reaction condition of step 4) is heating reflux for 12h in a closed device; And / or, the solvent in step 4) is acetonitrile, and the carbonate is sodium carbonate.

8. The production method according to claim 6, wherein In step 5), the solvent is trifluoroacetic acid or dichloromethane; And / or, the reaction condition of step 5) is room temperature reaction for 1h; And / or, in step 6), the solvent is dichloromethane; And / or, the reaction condition of step 6) is room temperature reaction for 1h; And / or, in step 7), the solvent is dichloromethane; And / or, the reaction condition of step 7) is room temperature reaction for 1h.

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