Michelia alba lactone derivative and application thereof

Abstract

The application belongs to the technical field of medicines, and particularly relates to a Michelia maudiae lactone derivative and application. The application provides a Michelia maudiae lactone derivative and application, which is a Michelia maudiae lactone derivative shown in the following formula (I) or a pharmaceutically acceptable salt thereof, and can be used for treating or repairing intestinal superficial mucosa damage caused by dietary stress. The Michelia maudiae lactone derivative is modified by a food-friendly hydrophilic side chain, has high oral availability, and meets food-grade safety. Formula (I)

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a micranthin derivative and its applications. Background Technology

[0002] Current research on the structural modification of Michelolone focuses entirely on the therapeutic fields of oncology and autoimmune diseases. Modification strategies aim to enhance cytotoxicity, anti-tumor activity, and potent anti-inflammatory efficacy. Existing modifications introduce highly active pharmacodynamic groups such as esters and amines, designed to kill diseased cells, but these exhibit non-selective cytotoxic side effects on normal intestinal epithelial cells, leading to insufficient long-term oral safety. Michelolone itself suffers from three inherent defects: poor water solubility, weak gastric acid stability, and low intestinal penetration. Current structural modification methods cannot simultaneously overcome these physicochemical and absorption barriers. Currently, there are no Michelolone derivatives, either domestically or internationally, that simultaneously possess food-grade safety, high water solubility, resistance to gastric acid degradation, and colon-targeted accumulation characteristics. There are also no Michelolone derivatives specifically targeting superficial intestinal mucosal damage caused by dietary stress (high-fat, spicy, late-night, etc.). Summary of the Invention

[0003] To address the problems of existing technologies, specifically targeting the repair of superficial inflammatory mucosal damage caused by dietary stress in the intestines, while overcoming the issues of poor water solubility, easy degradation by gastric acid, low intestinal epithelial penetration efficiency, and high toxicity of pharmaceutically modified derivatives of Michelolone, as well as its low oral bioavailability, limited dosage forms, and difficulty in targeting the colon, this invention provides a Michelolone derivative of formula (I), its stereoisomers, tautomers, isotope markers, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, or prodrugs: Formula (I) Where R1 represents H and C 1-7 Acyl group, C 1-6 alkyl.

[0004] In some implementation schemes, C 1-7 The acyl group is selected from formyl, acetyl, propionyl, butyryl, isobutyryl, chloroacetyl, and benzoyl. In some implementation schemes, C 1-6 The alkyl group is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopentyl, and cyclohexyl.

[0005] In some embodiments, the nitrobenzene derivative of formula (I) is selected from the following compounds:

[0006] The nitrobenzene lactone derivative of formula (I) of the present invention can be prepared by the following method: The compound shown in formula (I-1) reacts with 5-hydroxyuracil to give the compound shown in formula (I);

[0007] R1 has the definition described above.

[0008] In some embodiments, the preparation of the nitrifying lactone derivative shown in formula (I) is carried out in the presence of acetonitrile solvent and DBU.

[0009] In some embodiments, the preparation of the nitrosamine derivative shown in formula (I) is carried out at 20-30°C under nitrogen protection.

[0010] In some embodiments, the mass ratio of the compound shown in formula (I-1) to 5-hydroxyuracil is 1-3:1, such as 1:1, 2:1, 3:1.

[0011] The present invention also provides a pharmaceutical composition, food or health product comprising at least one of the following: a nitrosamine derivative of formula (I), its stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs.

[0012] According to embodiments of the present invention, the pharmaceutical composition, food, or health product further includes one, two, or more pharmaceutically / food-acceptable carriers or excipients.

[0013] According to the present invention, the pharmaceutical compositions and health products of the present invention can be formulated into dosage forms suitable for administration by methods known in the art.

[0014] When administered as a medicine, the compounds of the present invention can be given in pharmaceutical form. These medicines can be prepared in ways well known in the pharmaceutical field and can be given via a variety of routes, depending on whether local or systemic treatment is required and the area to be treated. Administration can be topically (e.g., transdermal, skin, eye, and mucous membrane delivery, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or blowing of powders or aerosols, including via nebulizers; intratracheal, intranasal), oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single large dose or via, for example, a continuous infusion pump. Topically administered pharmaceutical compositions and formulations may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional drug carriers, water, powdered or oily bases, thickeners, etc., may be necessary or required.

[0015] Some examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. Formulations may also contain: lubricants such as talc, magnesium stearate, and mineral oil; humectants; emulsifiers and suspending agents; preservatives such as methyl benzoate and hydroxypropyl benzoate; sweeteners and flavoring agents. The pharmaceuticals, foods, or health products of the present invention can be formulated using methods known in the art.

[0016] The tablets or pills of this invention can be coated or compounded to obtain dosage forms that provide the advantage of long-lasting action. For example, the tablets or pills contain an internal dose and an external dose component, the latter being a coated form of the former. The two components can be separated by an enteric coating layer, which is used to prevent disintegration in the stomach, allowing the internal component to pass through the duodenum intact or to delay release. A variety of substances can be used for such enteric coatings or coatings, including a variety of high molecular weight acids and mixtures of high molecular weight acids with such substances such as shellac, cetyl alcohol, and cellulose acetate.

[0017] The compounds and compositions of the present invention may be incorporated into liquid forms for oral or injectable administration, including aqueous solutions, suitably flavored syrups, water or oil suspensions; and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil; as well as elixirs and similar pharmaceutical solvents.

[0018] Compositions for inhalation or inhalation include solutions and suspensions, and powders, dissolved in pharmaceutically acceptable water or organic solvents or mixtures thereof. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered orally or via nasal inhalation to achieve local or systemic effects. The composition may be nebulized using an inert gas. The nebulized solution may be inhaled directly from a nebulizer, or the nebulizer may be connected to a face mask or intermittent positive pressure ventilation machine. Solutions, suspensions, or powder compositions may be administered orally or nasally by a device that delivers the formulation in a suitable manner.

[0019] The amount of compound or composition given to a patient is not fixed and depends on the drug being administered, the purpose of administration (e.g., prevention or treatment), the patient's condition, the route of administration, etc. In therapeutic applications, a sufficient amount of the composition may be given to a patient with an existing disease to cure or at least partially suppress the symptoms of the disease and its complications. The effective dose should depend on the disease state being treated and the judgment of the attending clinician, which depends on factors such as the severity of the disease, the patient's age, weight, and general condition.

[0020] The present invention also provides a method for treating or repairing superficial intestinal mucosal damage caused by dietary stress, comprising administering to a subject in need an effective amount of the nitrosamine derivative of formula (I), its stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs, or pharmaceutical compositions thereof, food, or health products.

[0021] The present invention also provides the use of the nitrosamine derivative of formula (I), its stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs in the preparation of medicaments, foods, and health products for treating or repairing dietary stress-induced superficial intestinal mucosal damage.

[0022] Beneficial effects The compounds represented by formula (I) of this invention, their stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts or prodrugs or pharmaceutical compositions thereof, foods, and health products can be used to treat or repair superficial intestinal mucosal damage caused by dietary stress. Through food-friendly hydrophilic side chain modification, they have high oral bioavailability, significantly improved resistance to degradation in the acidic digestive environment of the stomach, excellent intestinal penetration performance, targeted action on the colon, and meet food-grade safety requirements. Attached Figure Description

[0023] Figure 1 Compound 1 in Example 1 1 1H NMR (hydrogen nuclear magnetic resonance) spectrum; Figure 2 The results are LC-MS detection results for compound 1 in Example 1; Figure 3 The results of the effect of the experimental group in Example 6 on colonic IL-1β; Figure 4 The results of the effects of repeated drug administration on the pyroptosis pathway in colonic NLRP3 / Caspase-1 cells in the disease model animals in Example 6; Figure 5 This is a photograph of the colon tissue appearance of the experimental group in Example 6.

[0024] Terminology Definitions and Explanations Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures shall fall within the scope of this application specification.

[0025] Term "C" 1-6"Alkyl" should be understood to preferably represent a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0026] Term "C" 1-7 "Acyl" refers to an acyl functional group whose parent nucleus has a total of 1 to 7 carbon atoms. The general formula for acyl is RC (=O)-, where R is hydrogen and C is carbon. 1-6 Alkyl, substituted C 1-6 Alkyl or aryl, and the total number of carbon atoms of R and carbonyl carbon does not exceed 7; for example, formyl (HCO-), acetyl (CH3CO-), propionyl (CH3CH2CO-), butyryl (CH3CH2CH2CO-), isobutyryl ((CH3)2CHCO-), chloroacetyl (ClCH2CO-), benzoyl (PhCO-).

[0027] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of the specified compound's free acid and free base and has no adverse effects in biological or other respects. The compounds in this application also include pharmaceutically acceptable salts, such as nitrates, hydrochlorides, sulfates, or phosphates. A pharmaceutically acceptable salt is defined as a salt formed by converting a base group in the parent compound into its salt form. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts containing base groups such as amine (amino) groups. The pharmaceutically acceptable salts of this application can be synthesized from the parent compound by reacting a basic group in the parent compound with 1-4 equivalents of an acid in a solvent system.

[0028] The term "stereoisomer" refers to compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0029] The term "tautomer" refers to a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers possess different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated using high-resolution analytical techniques such as electrophoresis and chromatography, for example, HPLC.

[0030] The term "isotope-labeled" includes, but is not limited to, compounds of the present invention labeled with isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, sulfur, and chlorine (e.g., 2H, 3H, 13C, 14C, 15N, 18O, 17O, 18F, 35S, and 36Cl). Isotope-labeled compounds of the present invention can be used for the determination of the tissue distribution of the compounds, their prodrugs, and metabolites; preferred isotopes for such determinations include 3H and 14C. Furthermore, in some cases, substitution with a heavier isotope (e.g., deuterium (2H or D)) can provide increased metabolic stability, which offers therapeutic advantages, such as increased in vivo half-life or reduced dose requirements. The isotope-labeled compounds of the present invention can generally be prepared according to the methods described herein by replacing non-isotope-labeled reagents with isotope-labeled reagents.

[0031] The term "nitrogen oxide" refers to the oxidation of one or more nitrogen atoms to form N-oxides when a compound contains several amine functional groups. Specific examples of N-oxides are N-oxides of tertiary amines or N-oxides of nitrogen-containing heterocyclic nitrogen atoms. The corresponding amines can be treated with oxidizing agents such as hydrogen peroxide or peracids (e.g., peroxycarboxylic acids) to form N-oxides (see Advanced Organic Chemistry, Wiley Interscience, 4th ed., Jerry March, pages). In particular, N-oxides can be prepared using the LWDeady method (Syn.Comm. 1977, 7, 509-514), in which the amine compound is reacted with m-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as dichloromethane.

[0032] The term "solvent" refers to an association formed by one or more solvent molecules with the compounds of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed by solvent molecules that are water.

[0033] The term "metabolite" refers to the product obtained in vivo through the metabolism of a specific compound or its salt. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by experimental methods as described in this invention. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, acylation, deacylation, esterification, defatting, enzymatic cleavage, etc. Accordingly, this invention includes metabolites of compounds, including metabolites produced by sufficiently exposing the compounds of this invention to mammals for a period of time.

[0034] The term "ester" refers to an ester that can be hydrolyzed in vivo from compounds containing a hydroxyl or carboxyl group. Such esters are, for example, pharmaceutically acceptable esters that, upon hydrolysis in humans or animals, produce a parent alcohol or acid. The compounds of formula (I) of the present invention contain a carboxyl group and can form hydrolyzable esters in vivo with suitable groups, including, but not limited to, alkyl, arylalkyl, etc.

[0035] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred. Detailed Implementation

[0036] The technical solutions of this disclosure will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of this disclosure and should not be construed as limiting the scope of protection of this disclosure. All technologies implemented based on the above content of this disclosure are covered within the scope of protection intended by this disclosure.

[0037] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0038] Example 1: Preparation of Compound 1 Synthesis process route:

[0039] At 20-30℃, 1 kg of ternolactone was mixed with 7.32 kg of dichloromethane and stirred for 10-20 minutes until completely dissolved. After discharging, the reactor wall was rinsed with 0.66 kg of dichloromethane. The mixture of the feed liquid and the rinsing liquid was combined to obtain a ternolactone dichloromethane solution for later use. 14.63 kg of dichloromethane and 0.0255 kg of dichloromethane were then added to the reactor. kg of p-toluenesulfonic acid monohydrate was stirred and mixed at 20-30°C. Under a nitrogen atmosphere, the prepared solution was added dropwise, and the reaction was maintained at this temperature for 4-6 hours. The reaction endpoint was reached when the residual amount of ternolactone in the system was ≤5%, as monitored by HPLC. Then, a pre-prepared sodium bicarbonate aqueous solution was added to the reaction system, and the mixture was stirred at 20-30°C for 30-60 minutes. After standing for 20-40 minutes to separate the layers, the lower organic phase was collected. The resulting organic phase was washed sequentially with sodium bicarbonate aqueous solution and sodium chloride aqueous solution, each time with stirring at 20-30°C for 30-60 minutes, followed by standing and separation. After washing, 3.20 kg of anhydrous sodium sulfate was added to the organic phase, and the mixture was dried and stirred at 20-30°C for 30-60 minutes. The filtrate was collected by pressure filtration and then... The vessel wall and filter cake were washed with 1 kg of dichloromethane and filtered twice. All filtrates were combined and concentrated under reduced pressure at a temperature ≤40℃ until no obvious fractions were observed. The concentration was continued under reduced pressure for 1-2 hours to obtain concentrated material. 2.00 kg of ethyl acetate was added to the concentrated material and the temperature was raised to 70-80℃ to dissolve it completely. At the same temperature, 7.50 kg of n-heptane was slowly added dropwise. After the addition was completed, the temperature was lowered to -25~-15℃ and the mixture was stirred and crystallized for 1-3 hours. After crystallization, the liquid was removed by pressure filtration. The vessel wall and filter cake were washed with 1.36 kg of n-heptane and filtered again. The resulting filter cake was the wet product of Michelolactone. After HPLC analysis showed a purity ≥97.0%, the wet product was dried at 45-55℃ and a vacuum degree ≤-80 kPa for 3-10 hours, with the loss on drying controlled at LOD ≤1.5%. Michelolactone was finally obtained with a process molar yield of 65%.

[0040] Based on the actual amount of Michelia lactone added, 1 kg of Michelia lactone, 22.03 kg of acetonitrile, and 0.58 kg of pentazocine were mixed and dissolved by stirring at 20-30°C. Under nitrogen protection, 0.68 kg of DBU was added dropwise, and the reaction was maintained at this temperature for 3-5 hours. The reaction was terminated when the residual amount of Michelia lactone was ≤8% as determined by HPLC. The reaction mixture was then filtered, and the filter cake was washed with 0.63 kg of acetonitrile and the filtrate was collected and stored for later use. 0.49 kg of Michelia lactone was prepared beforehand. 36% hydrochloric acid was mixed with 4.51 kg of purified water to prepare a hydrochloric acid aqueous solution. This solution was added dropwise to the filtrate at -5 to 5°C to adjust the pH to 6.0 to 7.0. The mixture was stirred for 1 to 3 hours, followed by pressure filtration. The filter cake was washed with 0.78 kg of acetonitrile to obtain a wet product of compound 1. This wet product underwent two purification processes with pure water and one purification process with anhydrous ethanol, each time stirred at 20 to 30°C for 1.5 to 2.5 hours. After filtration, the filter cake was washed with anhydrous ethanol. The purity of the obtained wet product was ≥98.0% as determined by HPLC. Finally, the wet product was dried at 45 to 55°C under a vacuum of ≤-80 kPa for 3 to 12 hours, with a loss on drying (LOD) of ≤1.0%. After successful drying, compound 1 was obtained as the final product, with a process molar yield of 50%. 1 The results of 1H NMR and LC-MS detection are as follows: Figure 1 , 2 As shown.

[0041] Example 2: Simulated Gastric Fluid Stability Test Example 1 Preparation of Compound 1: Purity: 99.2%, organic solvent residue conforms to GB 31640; Michelolone reference standard: purity: 98%.

[0042] The simulated gastric juice was prepared according to the standard of pH 1.2, containing 10 g / L pepsin, and according to the General Chapter 0931 of Part IV of the 2025 edition of the Chinese Pharmacopoeia. It was destroyed at a constant temperature of 37℃. 0.05 g each of Compound 1 prepared in Example 1 and sulfadiazine reference standard were placed in a dissolution vessel containing 900 mL of simulated gastric juice at 37℃. The mixture was continuously shaken at 50 rpm for 2 h. Immediately afterward, 1 mL of the solution was taken, and the pH was adjusted to neutral with 0.1 mol / L sodium hydroxide solution. The solution was then diluted to volume with the mobile phase, filtered through a 0.22 μm filter membrane, and injected. The contents of Compound 1 and sulfadiazine were determined by high performance liquid chromatography (HPLC). The gastric juice retention rate was calculated using the formula Retention Rate (%) = (Ct / C0) × 100, where Ct is the concentration measured after 2 h of gastric juice treatment, and C0 is the original content of Compound 1 and sulfadiazine. The results are shown in Table 1.

[0043] Table 1: Retention rates of compound 1 and sulforaphane in simulated gastric juice

[0044] Table 1 shows that the remaining contents of the two groups of samples at the initial 0 h were 98% and 99.2%, respectively. With increasing time, the degradation rate of sulfadiazine was significantly higher at 0.5 h, 1 h, and 2 h. After 2 h, only 27.1% of sulfadiazine remained, while the derivative of this invention maintained a higher retention rate of 87.6%. Compared to sulfadiazine, the stability of compound 1 increased by 3.2 times after 2 h of degradation. These results indicate that the derivative of this invention has significantly improved resistance to degradation in the acidic digestive environment of the stomach, effectively reducing degradation losses in the stomach after oral administration, and is more suitable for the development of oral dietary supplements.

[0045] Example 3: Solubility Test Referring to the general requirements for solubility determination in the General Provisions of Part IV of the 2025 edition of the Chinese Pharmacopoeia, the equilibrium method was used for the experiment, with the test temperature kept constant at 25℃. Excess amounts of sulfadiazine reference standard and compound 1 prepared in Example 1 were added to an appropriate amount of pure water and placed in a constant-temperature shaker for continuous shaking until dissolution equilibrium was reached (samples were taken after 24 hours of shaking; if the deviation between two consecutive measurements was less than 2%, equilibrium was considered reached). After standing and separation, the upper saturated clear solution was taken, filtered through a 0.22 μm filter membrane, and the contents of sulfadiazine and compound 1 in the saturated solution were determined by high-performance liquid chromatography (external standard method), and the corresponding solubilities were calculated. The test showed that under pure water conditions at 25℃, the solubility of sulfadiazine was 0.08 mg / mL; the solubility of compound 1 was 1.54 mg / mL. Compared with sulfadiazine, the solubility of compound 1 increased by 19.2 times, indicating that the water solubility of compound 1 prepared in this invention was significantly improved.

[0046] Example 4: Caco-2 cell bidirectional permeability assay Caco-2 cells were seeded into 96-well Transwell cell culture plates and cultured continuously for 21 days before being used for a permeability assay. Fibrolactone, 5-hydroxyuracil, and Compound 1 were administered bidirectionally to both sides of the Transwell cell culture plate at a dietary intake concentration of 1.00 µM. After incubation for 120 min, samples were collected, and the concentrations of Fibrolactone, Compound 1, and 5-hydroxyuracil in the transport medium were determined using LC-MS / MS. The Papp values ​​for bidirectional transport were calculated, and the results are shown in Table 2.

[0047] Table 2: Results of bidirectional transport of sulfadiazine, 5-hydroxyuracil, and compound 1 via Papp

[0048] The results showed that, under the same dietary intake concentration of 1.00 µM, the intestinal epithelial absorption and transport capacity of the derivative of this invention was significantly improved compared with sulfadiazine and 5-hydroxyuracil. The apparent permeability coefficient of compound 1 in the absorption direction from the apex to the basal side (A→B) was 4.3 times that of sulfadiazine, and the transepithelial absorption efficiency of the intestine was greatly improved. The transport capacity in the transepithelial outflow direction (B→A) was 5.8 times that of sulfadiazine. 5-hydroxyuracil is a compound with extremely low intestinal permeability and extremely poor transepithelial absorption capacity. After molecular construction, this permeability can be greatly improved. The derivative compound is more likely to penetrate intestinal epithelial tissue and has excellent intestinal penetration performance. It can effectively increase the amount of intestinal tissue absorbed after oral dietary intake and is suitable for the development of oral foods for intestinal targeted maintenance.

[0049] Example 5: Oral absorption and distribution in healthy SD rats SD rats: Fasted for at least 12 hours before administration, with free access to water, and fed uniformly 4 hours after administration.

[0050] Twenty-six SD rats were randomly divided into four groups according to their body weight: G1, G2, and G3, with six rats in each group (half male and half female). Detailed grouping information is shown in Table 3 below. Groups G1, G2, and G3 were administered via gavage at a dose of 5 mg / kg.

[0051] Table 3: Grouping and Dosage

[0052] Note: F represents female, M represents male; Rats were administered the drug by gavage, and blood samples were collected at the time points specified in Table 4. Blood was collected via the jugular vein or other suitable veins, and K2-EDTA was used for anticoagulation. Plasma was obtained by centrifugation within 1 hour after blood collection. The test samples were stored at -80ºC before analysis. The results of in vivo pharmacokinetic tests are shown in Table 4.

[0053] Table 4: Pharmacokinetic Test Results: ( ±SD, n=3)

[0054] Example 6: Dietary repair of rat intestines and tissue pharmacokinetics test Experimental animals: SD rats, SPF grade, sourced from Sichuan Vitonlihua Experimental Animal Technology Co., Ltd., 16 rats per group, 8 males and 8 females.

[0055] Since dietary stress-induced inflammatory mucosal damage in the intestines is caused by exogenous dietary factors such as high-fat, spicy, alcoholic, overeating, irregular sleep patterns, and dietary irritants, it is difficult to effectively construct a rat model using TNBS solvent alone or simply by using ethanol, high-fat, or spicy solutions alone. This experiment used TNBS and ethanol enema to construct a rat model of dietary stress-induced superficial intestinal mucosal damage, simulating reversible superficial intestinal damage induced by dietary irritants. From day 1 (D1) after model establishment, daily oral gavage intervention was initiated, once daily for 7 consecutive days.

[0056] The experimental groups were set as follows: normal control group, model control group, sulfadiazine dietary intervention group (administered dose 6 mg / kg), 5-hydroxyuracil dietary intervention group (administered dose 6 mg / kg), low-dose group of the derivative of this invention (compound 1) (administered dose 1 mg / kg), medium-dose group of the derivative of this invention (compound 1) (administered dose 3 mg / kg), and high-dose group of the derivative of this invention (compound 1) (administered dose 6 mg / kg). All groups were administered the derivative by gavage once daily for 7 consecutive days. The administration solvent was 20% hydroxypropyl-β-cyclodextrin aqueous solution, with a uniform administration volume of 10 mL / kg.

[0057] The expression level of IL-1β protein in animal colon tissue was detected by Western blotting. Specific detection conditions and procedures are as follows: Colon tissue samples were collected from all test animals, and detection was performed uniformly after all samples were collected. Colon tissue samples were lysed with tissue lysis buffer containing protease inhibitors and phosphatase inhibitors to extract total protein. The concentration of the extracted total protein was quantified using the BCA protein quantification method, and a standardized protein loading buffer was prepared based on the quantification results.

[0058] Western blotting was used to specifically detect the expression levels of IL-1β protein and activated cleaved IL-1β protein in the samples. GAPDH or Tubulin was used as internal control proteins to correct for sample loading errors. Finally, the relative expression level of IL-1β protein was calculated by grayscale analysis, thus realizing the quantitative detection of IL-1β content in colon tissue.

[0059] The results of the above experimental groups on the effect of colonic IL-1β are as follows: Figure 3 As shown, the results indicate that, under the same dosage of 6 mg / kg for 7 days, the derivative of this invention has a significantly better regulatory effect on the low-stress pro-inflammatory factor IL-1β in the colon than the sulfadiazine group and the 5-hydroxypyrimidine group, and has outstanding efficacy in repairing and regulating intestinal dietary damage.

[0060] To further clarify the effect of the derivative of this invention (compound 1) on the expression of activated Caspase-1 and activated IL-1β effector proteins in colon tissue, 1 mg / kg and 6 mg / kg were administered by gavage once daily for 7 consecutive days.

[0061] Colon tissue was collected from SD rats. After all samples were collected, they were tested uniformly. All colon tissues were lysed and total protein was extracted using tissue lysis buffer (containing protease and phosphatase inhibitors). Protein loading buffer was prepared after BCA method for protein quantification. Western blotting was used for detection. The target proteins were downstream signaling proteins of inflammasome 3 (NLRP3). GAPDH or Tubulin was used as internal control to analyze the relative expression level of the target proteins.

[0062] The experimental results of the normal control group, model control group, low-dose group (administered dose 1 mg / kg) of the derivative of this invention (compound 1), and high-dose group (administered dose 6 mg / kg) of the derivative of this invention (compound 1) after drug administration are as follows: Figure 4 As shown in the figure (the results are data obtained by combining male and female rats), the results indicate that the relevant indicators decrease in a gradient with increasing administration dose, and the steady-state regulation effect has a clear dose-dependent characteristic.

[0063] Rats were administered the following drugs by gavage once daily for 7 consecutive days: a control group of micranthin (6 mg / kg), a control group of 5-hydroxyuracil (6 mg / kg), a control group of micranthin + 5-hydroxyuracil (3 mg / kg each of micranthin and 5-hydroxyuracil), and a high-dose group of the derivative of this invention (compound 1) (6 mg / kg). After administration, the rats were weighed and their weight recorded. They were anesthetized with 10% urethane, fixed on a rat board, and their abdomens were cut open to collect blood. The large and small intestines were then separated, the colon length was measured, and the degree of intestinal edema and weight gain due to stress was observed. The results are as follows: Figure 5 As shown (the results are data obtained by combining male and female rats).

[0064] The results showed that the high-dose group of the derivative of the present invention (compound 1) could significantly increase the length of colon tissue damaged by dietary stress, reduce the weight gain of intestinal tissue edema caused by intestinal stress, improve the macroscopic morphology of damaged colon in model rats (n=8), and achieve the repair and remodeling of the superficial mucosal tissue structure of the intestine. Its therapeutic effect was significantly greater than that of the sulfadiazine control group and the 5-hydroxyuracil control group.

[0065] The above description provides an exemplary account of the implementation methods of the technical solution disclosed herein. It should be understood that the scope of protection of this disclosure is not limited to the above-described embodiments. Any modifications, equivalent substitutions, or improvements made by those skilled in the art within the spirit and principles of this disclosure should be included within the scope of protection of the claims of this application.

Claims

1. A micranthin derivative or a pharmaceutically acceptable salt thereof, characterized in that, The structure of the Michelia lactone derivative is shown in formula (I): Formula (I) Where R1 represents H and C 1-7 Acyl group, C 1-6 alkyl.

2. The nitrobenzene lactone derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, R1 is selected from formyl, acetyl, propionyl, butyryl, isobutyryl, chloroacetyl, and benzoyl.

3. The micranthin derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, R1 is selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopentyl, and cyclohexyl.

4. The micranthin derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, Michelia lactone derivatives are selected from the following compounds: 。 5. A method for preparing the micranthin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that, Includes the following steps: The compound shown in formula (I-1) reacts with 5-hydroxyuracil to give the compound shown in formula (I); Wherein, R1 has the definition as described in any one of claims 1 to 4.

6. The method for preparing the nitrobenzin derivative or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that, The preparation of the nitrifying lactone derivative shown in formula (I) was carried out in the presence of acetonitrile solvent and DBU.

7. The method for preparing the nitrobenzin derivative or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that, The preparation of the nitrosamine derivative shown in formula (I) was carried out at 20-30°C under nitrogen protection.

8. The method for preparing the nitrobenzin derivative or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that, The mass ratio of the compound shown in formula (I-1) to 5-hydroxyuracil is 1-3:

1.

9. A pharmaceutical composition, food, or health product comprising at least one of the nitrosamine derivatives of any one of claims 1-4 or a pharmaceutically acceptable salt thereof.

10. Use of the nitrobenzene derivative of any one of claims 1-4 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or repairing dietary stress-induced superficial intestinal mucosal damage.

Images