Pharmaceutical Uses of Tropical Acid and its Derivatives in the Preparation of Drugs for the Treatment of Immune and Inflammation-Related Diseases

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to the use of tropic acid and its derivatives in the preparation of drugs for treating immune and inflammation-related diseases, and to drugs for treating immune and inflammation-related diseases. The tropic acid and its derivatives are compounds of formulas I-IV or pharmaceutically acceptable salts thereof, as well as solvent compounds, enantiomers, diastereomers, tautomers, or mixtures thereof in any proportion, including racemic mixtures, of the compounds of formulas I-IV or pharmaceutically acceptable salts thereof. Animal studies show that tropic acid and its derivatives have broad-spectrum immunomodulatory, anti-inflammatory, and analgesic effects, significantly improving pathological indicators in numerous animal models of immune and inflammation-related diseases. They can regulate abnormal immune responses towards normalcy and exert good anti-inflammatory and analgesic effects, making them suitable for patients with different disease degrees and types of immune and inflammation-related diseases, and thus have industrial applications.

Description

[0001] This application is a divisional application of the invention patent application filed on March 8, 2023, with application number 202310217736.3 and invention title "Pharmaceutical use of tropinic acid and its derivatives in the preparation of drugs for the treatment of immune and inflammatory diseases". Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, and in particular relates to the use of tropinic acid and its derivatives in the preparation of drugs for the treatment of immune and inflammatory diseases, as well as drugs for the treatment of immune and inflammatory diseases. Background Technology

[0003] Immunological diseases are a large category of diseases characterized by localized or systemic abnormal inflammatory immune responses, mainly including hypersensitivity reactions, immunodeficiency diseases, and autoimmune diseases. Type I hypersensitivity reactions include penicillin allergy, drug-induced rashes, seasonal, pollen, or dust-induced allergic rhinitis, pharyngitis, conjunctivitis, bronchial asthma, eczema, and urticaria; Type II hypersensitivity reactions include neonatal hemolytic reactions, drug-induced hemolytic anemia, and aplastic anemia; Type III hypersensitivity reactions include glomerulonephritis; and Type IV hypersensitivity reactions include tuberculosis and syphilis. Infection-related diseases include bronchitis or pneumonia, gastroenteritis, endometritis, otitis media, tonsillitis, boils, sinusitis, abscesses or granulomas, sepsis, septicemia, myocarditis, meningitis, osteoarthritis, pleurisy, cholecystitis, osteomyelitis, prostatitis, urethritis, cystitis, proctitis, paronychia, and folliculitis. Autoimmune diseases include hepatitis, systemic lupus erythematosus, spondylitis, rheumatoid arthritis, nephritis, diabetes, pancreatitis, enteritis, rheumatic heart disease, pneumonia, scleroderma, vasculitis, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, and autoimmune thyroiditis. It is estimated that the incidence of autoimmune diseases is increasing year by year, with approximately 7.6% to 9.4% of the global population suffering from various types of autoimmune diseases. These diseases are difficult to cure, and most patients require long-term or even lifelong medication. Some diseases, such as lupus nephropathy, are extremely dangerous, severely impacting patients' quality of life and threatening their lives. Approximately 50 million Americans (about one-fifth of the total population) suffer from autoimmune diseases, of which about 75% are women. Autoimmune diseases have become the third leading cause of chronic disease after cardiovascular disease and cancer. Although there is currently no definitive incidence data in China, the patient population is increasing annually.

[0004] Treatment of autoimmune diseases has two main goals: symptom relief and functional maintenance, and slowing the progression of tissue damage. Currently, drugs for treating autoimmune diseases are mainly classified into nonsteroidal anti-inflammatory drugs (NSAIDs), steroidal anti-inflammatory drugs (SAIDs), disease-modifying antirheumatic drugs (DMARDs), biologics, and natural medicines. NSAIDs are commonly used to treat autoimmune diseases, effectively reducing clinical symptoms and signs and eliminating local inflammatory responses. However, these drugs cannot control disease progression, and common adverse reactions include central nervous system symptoms, cardiovascular damage, gastrointestinal symptoms, hematopoietic system changes, liver and kidney dysfunction, asthma, and drug eruptions. SAIDs have strong anti-inflammatory and immunosuppressive effects, preventing inflammatory cells from accumulating at the site of inflammation, inhibiting the release of inflammatory factors, and suppressing the proliferation and secretion of TB lymphocytes. These drugs have many adverse reactions, and relapse is common after discontinuation. Currently, they are often used in combination with other immunosuppressants in clinical practice. DMARDs are widely used in the treatment of autoimmune diseases such as chronic kidney disease, transplant rejection, and tumors. Although the chemical structures and mechanisms of action of traditional DMARDs are not entirely the same, their clinical pharmacodynamic characteristics are similar: slow onset of action, with symptoms and signs gradually alleviating after several weeks or months of use. Long-term continuous use can achieve relatively stable efficacy. The main adverse reactions include gastrointestinal reactions, bone marrow suppression, infection, and liver and kidney damage. Biologics exert their therapeutic effects by blocking key inflammatory cytokines or cell surface molecules, such as monoclonal antibodies targeting IL-1, IL-6, TNF-α, and IL-17, anti-CD20 monoclonal antibodies, B lymphocyte-stimulating factor (BAFF) inhibitors, T-cell inhibitors, integrin monoclonal antibodies, and selective adhesion molecule inhibitors. Most of these drugs are in the clinical trial stage, and a few are already on the market. They have many and serious adverse reactions, and some drugs have been banned due to serious adverse reactions. Natural drugs used to treat immune diseases include glycosides and alkaloids. Glycosides include total glucosides of paeony, total glucosides of ginseng, total glucosides of gynostemma pentaphyllum, astragaloside A, total glucosides of triptolide, and total saponins of Panax notoginseng. Alkaloids include sinomenine, total alkaloids of aconite, sophoridine, and triptolide. These drugs have fewer adverse reactions and mostly have anti-inflammatory, analgesic, and immunosuppressive effects, but their clinical treatment is not highly targeted and the efficacy is not ideal. With the deepening understanding of the pathological mechanisms of immune diseases and the discovery of new drug targets, in addition to NSAIDs, SAIDs, and traditional DMARDs, targeted small molecule drugs such as tofacitinib, baricitinib, upatacitinib, and filgotinib have also been developed and applied clinically to treat inflammatory and immune diseases. These drugs have definite efficacy, but they also have adverse reactions such as gastrointestinal symptoms, immunosuppression, bone marrow suppression, infection, and new tumor formation. Therefore, developing small molecule drugs with immunomodulatory and anti-inflammatory effects without impairing the body's physiological functions is the main strategy and direction for treating immune and inflammation-related diseases.

[0005] In summary, the patient population with immune and inflammatory diseases is large. Although there are many types of drugs or methods for treatment, their efficacy is limited, often requiring long-term, repeated, or even lifelong treatment. Some drugs may achieve temporary relief, but the short-term relapse rate is high. Most drugs are often limited by their inherent toxicity and selectivity, and various systemic and local adverse reactions are unavoidable. To address these issues, there is an urgent need to develop therapeutic drugs that are fast-acting, have short treatment courses, require small dosages, have few adverse reactions, low relapse rates, are inexpensive, and are easy to use. Simultaneously, the development of drug formulations such as topical, oral, and injectable formulations should be considered to suit patients with different disease severities and types. This invention, through extensive animal experiments, screened and clarified the effectiveness of tropic acid (DL-Tropic Acid, also known as 2-phenyl-3-hydroxypropionic acid) and its derivatives in treating immune and inflammatory diseases. Tropic acid is an intermediate in the synthesis of atropine, and the raw material is inexpensive. Currently, there are no reports on the prevention and treatment of immune and inflammatory diseases using tropic acid and its derivatives. Summary of the Invention

[0006] This invention provides, in one aspect, the use of tropinic acid and its derivatives, pharmaceutically acceptable salts thereof, solvent compounds, enantiomers, diastereomers, tautomers, or mixtures thereof in any proportion thereof in the preparation of a formulation for the prevention and / or treatment of immune and inflammatory-related diseases, wherein the tropinic acid and its derivatives have the structure shown in Formula A: R1-R5 are each independently selected from -H, -OH or C1-C6 alkoxy groups.

[0007] In one implementation, R1-R5 are each independently selected from -H or -OH.

[0008] In one implementation, R1 and R5 are selected from -H, and R2-R4 are each independently selected from -H or -OH.

[0009] In one embodiment, R1 and R5 are selected from -H, two or three of R2-R4 are selected from -OH, and the rest are selected from -H.

[0010] In one implementation, at least two of R1-R5 are selected from -OH.

[0011] In one implementation, two or three of R1-R5 are selected from -OH.

[0012] In one implementation, two or three of R2-R4 are selected from -OH.

[0013] In one embodiment, the tropinic acid and its derivatives are selected from compounds represented by formulas I-IV: .

[0014] Wherein, Formula I is DL-Tropic Acid, Formula II is 4-Hydroxy-α-(hydroxymethyl)benzeneacetic acid, Formula III is 3,4-Dihydroxy-α-(hydroxymethyl)benzeneacetic acid, and Formula IV is 3,4,5-Trihydroxy-α-(hydroxymethyl)benzeneacetic acid.

[0015] The tropinic acid and its derivatives described in this invention have immunomodulatory, anti-inflammatory and analgesic effects. They have a significant effect on improving the pathological indicators of many animal models of immune and inflammation-related diseases, can regulate abnormal immune responses to normal, and exert good anti-inflammatory and analgesic effects.

[0016] In one embodiment, the immune and inflammation-related diseases are selected from one or more of the following: allergic rhinitis, bronchitis, bronchial asthma, pharyngitis, conjunctivitis, eczema, urticaria, neonatal hemolytic reaction, hemolytic anemia, aplastic anemia, nephritis, tuberculosis, syphilis, pneumonia, gastroenteritis, endometritis, otitis media, sepsis, septicemia, myocarditis, meningitis, tonsillitis, sinusitis, pleurisy, cholecystitis, osteomyelitis, prostatitis, urethritis, cystitis, anorectal inflammation, paronychia and folliculitis, osteoarthritis, hepatitis, systemic lupus erythematosus, spondylitis, rheumatoid arthritis, diabetes, pancreatitis, enteritis, rheumatic heart disease, vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, and thyroiditis.

[0017] A second aspect of the present invention provides a medicament for the prevention and / or treatment of immune and inflammation-related diseases, the medicament comprising tropinic acid and its derivatives, pharmaceutically acceptable salts thereof, solvent compounds, enantiomers, diastereomers, tautomers or mixtures thereof in any proportion thereof.

[0018] The drug provided by this invention for the prevention and / or treatment of immune and inflammatory-related diseases has broad-spectrum immunomodulatory, anti-inflammatory and analgesic effects, and is highly effective.

[0019] In one embodiment, tropinic acid and its derivatives, pharmaceutically acceptable salts thereof, solvent compounds, enantiomers, diastereomers, tautomers, or mixtures thereof in any proportion thereof serve as the active ingredient in the medicament of the present invention. Preferably, it serves as the principal active ingredient; more preferably, it serves as the sole active ingredient.

[0020] In the above-mentioned uses and drugs, tropinic acid and its derivatives, pharmaceutically acceptable salts, solvent compounds, enantiomers, diastereomers, tautomers or mixtures thereof in any proportion can be prepared with pharmaceutically acceptable carriers or excipients into drug dosage forms for external, oral or injectable administration.

[0021] Therefore, in this invention, the drug can be a topical drug, an oral drug, or an injectable drug.

[0022] In this invention, the drug may contain a pharmaceutically acceptable carrier or excipient. The drug can be formulated into various conventional solid, liquid, or semi-solid dosage forms, such as granules, tablets, or capsules; liquid dosage forms such as sprays and injections; and semi-solid dosage forms such as creams. In one aspect, the dosage form of the drug may be: powder, tablet, coated tablet, granule, capsule, solution, emulsion, suspension, injection, spray, nasal spray, aerosol, powder spray, lotion, liniment, ointment, plaster, paste, gel, patch, etc.

[0023] In this invention, the term "pharmaceutically acceptable carrier or excipient" includes any and all solvents, cosolvents, dispersion media, coating materials, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delay agents, salts, preservatives, drug stabilizers, binders, excipients, diluents, flow aids, granulators, disintegrants, thickeners, viscous agents, lubricants, anti-caking agents, humectants, wetting agents, chelating agents, plasticizers, dyes, flavoring agents, etc., and combinations thereof, as is well known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 19th Ed. Mack Printing Company, 1995; Shanghai Pharmaceutical Industry Research Institute et al., Pharmaceutical Excipient Application Technology (Second Edition), China Medical Science and Technology Press, 2002; Comparative Handbook of Pharmaceutical Excipient Standards of Various Countries, Volumes 1-3, compiled by the National Pharmacopoeia Commission, China Medical Science and Technology Press, 2016; Handbook of Pharmaceutical Excipients, Raymond C. Rowe, PJ). (Paul J. Sheskey, Paul J. Weller, eds., translated by Zheng Junmin, Chemical Industry Press, 2005, etc.). Except for carriers and excipients incompatible with the active ingredient, any conventional carriers and excipients may be considered in therapeutic or pharmaceutical compositions.

[0024] For example, as a solid dosage form, the pharmaceutically acceptable carrier or excipient may include at least one of the following: (a) fillers such as starch, corn starch, modified starch, compressible starch, lactose, lactose monohydrate, microcrystalline cellulose, cyclodextrin, sorbitol, mannitol, calcium phosphate, amino acids, etc.; (b) binders such as starch paste, gelatinized starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, low-substituted hydroxypropyl cellulose, polyvinylpyrrolidone, gelatin, alginate, etc.; (c) humectants such as glycerin; (d) disintegrants such as dry starch, modified starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone carboxymethyl cellulose, microcrystalline cellulose, effervescent disintegrants, crospovidone, etc.; (e) solution blockers such as paraffin; (f) absorption enhancers such as quaternary ammonium compounds; (g) Wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) absorbents, such as kaolin and bentonite; (i) lubricants, such as talc, stearic acid, magnesium or calcium stearate, micronized silica gel, hydrogenated castor oil and solid polyethylene glycol, polyethylene glycol 4000-20000, magnesium dodecyl sulfate, etc.

[0025] In this invention, the drug is applicable to humans or other warm-blooded animals. When applicable to humans, the preferred dosage of tropinic acid and its derivatives, either alone or in combination, is 1 mg / kg·d to 50 mg / kg·d, more preferably 10 mg / kg·d to 20 mg / kg·d. The therapeutically effective amount of the compound or pharmaceutical composition depends on the individual's species, weight, age, individual condition, the disease being treated, or its severity. Physicians, clinicians, or veterinarians with common skills can easily determine the effective amount of each active ingredient required for the prevention, treatment, or inhibition of disease progression.

[0026] The present invention also provides a compound of formula A, a pharmaceutically acceptable salt thereof, a solvent compound, an enantiomer, a diastereomer, a tautomer, or a mixture thereof in any proportion: Among them, R1-R5 are each independently selected from -H or -OH; The condition is that at least two of R1-R5 are selected from -OH.

[0027] In one implementation, two or three of R1-R5 are selected from -OH.

[0028] In one implementation, two or three of R2-R4 are selected from -OH.

[0029] In one embodiment, the compound is selected from the compounds shown in Formulas III-IV: .

[0030] Pharmaceutically acceptable salts of the compounds of this invention include their base addition salts and acid addition salts. Preferably, the base addition salts are selected from sodium, potassium, calcium, lithium, magnesium, zinc, ammonium, tetramethylammonium, tetraethylammonium, triethylamine, trimethylammonium, ethylamine, diethanolamine, arginine, or lysine salts; the acid addition salts are selected from organic acid salts such as acetate, aspartate, benzoate, benzenesulfonate, citrate, ethanedisulfonate, ethanesulfonate, formate, fumarate, gluconate, glucuronate, lactate, malate, trifluoroacetate, and maleate, as well as inorganic acid salts such as hydrochloride, hydrobromide, hydrogen sulfate, nitrate, and phosphate. The free form of the compounds of this invention can be converted into the corresponding salt form; and vice versa. The free or salt form and solvated form of the compounds of this invention can be converted into the non-solvated free or salt form of the corresponding compounds; and vice versa.

[0031] The compounds of the present invention also comprise their solvated forms, which refer to associative compounds formed by one or more solvent molecules with the compounds of the present invention. Solvents forming solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol.

[0032] The compounds of the present invention can exist as isomers and mixtures thereof; for example, tautomers, optical isomers, enantiomers, and diastereomers. The compounds of the present invention may, for example, contain an asymmetric carbon atom, and therefore can exist as enantiomers or diastereomers and mixtures thereof, for example, as racemates. The compounds of the present invention can exist in (R)-, (S)-, or (R, S)- configurations, preferably in the (R)- or (S)- configuration at specific positions in the compound.

[0033] The present invention has the following advantages and effects compared with the prior art: (1) This invention is the first to discover that tropinic acid and its derivatives can significantly improve pathological indicators in animal models of immune and inflammation-related diseases; (2) Tropical acid and its derivatives can regulate abnormal immune responses to normalization and exert good anti-inflammatory and analgesic effects. (3) Tropical acid and its derivatives are the main components for treating immune and inflammatory diseases. Compared with existing drugs, they have the advantages of low toxicity, fast onset of action, short course of treatment, small dosage, low recurrence rate and convenient use. They also take into account external, oral and injection dosage forms, and can be adapted to patients with different disease degrees and different types of immune and inflammatory diseases. (4) The tropinic acid derivative compounds 3 and 4 obtained through structural modification have better and more significant therapeutic effects than tropinic acid in the treatment of immune and inflammation-related diseases; (5) Tropical acid and its derivatives used in this invention are easy to obtain and synthesize, inexpensive, stable, easy to store and transport, and suitable for industrial application. Attached Figure Description

[0034] Figure 1 Compounds of Formula III 1 H-NMR spectrum Figure 2 Compounds of Formula III 13 C-NMR spectrum Figure 3 Compounds of formula IV 1 H-NMR spectrum Figure 4 Compounds of formula IV 13 C-NMR spectrum Figure 5 Effects of tropinic acid and its derivatives on liver histopathology in a mouse model of autoimmune hepatitis (x200) Figure 6 Effects of tropinic acid and its derivatives on spleen histopathology in a mouse model of autoimmune hepatitis (x200) Figure 7 Effects of tropinic acid and its derivatives on lung tissue pathology in a mouse model of pulmonary inflammation caused by novel coronavirus (x200) Figure 8 Comparative observation of morphological differences in ankle joints and paws before and after treatment in rats of different groups. Figure 9 Comparison of trends in joint index and swelling in rats of different groups . Detailed Implementation

[0035] The preferred embodiments of the present invention will be described below with reference to the accompanying drawings. It should be understood that the preferred embodiments described herein are for illustration and explanation only and are not intended to limit the present invention. The described embodiments are merely some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Example 1: Preparation of compound III (3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid) 1. Synthesis process Under ice-water bath conditions, 1.96 g (10 mmol, 1.0 eq.) of 3,4-dimethoxyphenylacetic acid was placed in a 100 mL Schlenk flask. Under nitrogen protection, 20 mL of anhydrous DCM was added, and 1.8 mL (25 mmol, 2.5 eq.) of thionyl chloride was slowly added dropwise while maintaining low temperature. The system was then transferred to room temperature and stirred for 2 h. The solvent and excess thionyl chloride were removed under reduced pressure at 40 °C. Subsequently, 20 mL of methanol was added to the system and stirred overnight at room temperature. The solvent was removed by distillation under reduced pressure to obtain 1.50 g of methyl 3,4-dimethoxyphenylacetic acid.

[0037] At room temperature, 1.0 g (4.76 mmol, 1.0 eq.) of methyl 3,4-dimethoxyphenylacetate was placed in a 100 mL Schlenk flask, along with 0.171 g (5.71 mmol, 1.2 eq.) of paraformaldehyde and 5 mL of DMSO. Under nitrogen protection, 0.0257 g (0.476 mmol, 10 mol%) of sodium methoxide was added as a catalyst, and the mixture was stirred overnight at room temperature. For post-treatment, the reaction mixture was transferred to 100 mL of water, and the aqueous layer was extracted with EtOAc. 20 mL of organic phase was combined, washed with water and dried, concentrated under reduced pressure and purified by column chromatography to obtain 0.90 g of methyl 3,4-dimethoxy-α-(hydroxymethyl)phenylacetate.

[0038] 0.6 g (2.5 mmol, 1.0 eq.) of methyl 3,4-dimethoxy-α-(hydroxymethyl)phenylacetate was placed in a 20 mL Schlenk flask. 2 mL of 40% hydrogen bromide solution was slowly added to the system at room temperature, and the mixture was heated under reflux overnight. For post-treatment, the reaction mixture was transferred to 50 mL of water, and the aqueous layer was extracted using DCM. 20 mL of organic phase was combined, washed with water and dried, concentrated under reduced pressure and purified by column chromatography to obtain 0.35 g of 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid.

[0039] 2. Structural Identification 3,4-Dihydroxy-α-(hydroxymethyl)phenylacetic acid is a white crystalline powder, readily soluble in methanol and soluble in water. 1 H-NMR (400 MHz, CD3OD) See Figure 1 , δ H (ppm) 3.63 (2H, m, CH2), 3.99 (1H, m, CH), 6.50(1H, dd, J =2.0, 8.0 Hz, Ph-H), 7.04 (1H, d, J= 8.0 Hz, Ph-H), 7.09 (1H, d, J =2.0 Hz, Ph-H); 13 C-NMR (400 MHz, CD3OD) See Figure 2 , δ C (ppm) 55.9 (CH), 65.2 (CH2), 114.3 (CH), 116.4 (CH), 123.9 (CH), 127.2 (C), 145.3 (C), 146.0 (C), 176.2 (C=O).

[0040] Example 2: Preparation of compound IV (3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid) 1. Synthesis process Under ice-water bath conditions, 2.26 g (10 mmol, 1.0 eq.) of 3,4,5-trimethoxyphenylacetic acid was placed in a 100 mL Schlenk flask. Under nitrogen protection, 20 mL of anhydrous DCM was added, and 1.8 mL (25 mmol, 2.5 eq.) of thionyl chloride was slowly added dropwise while maintaining low temperature. The system was then transferred to room temperature and stirred for 2 h. The solvent and excess thionyl chloride were removed under reduced pressure at 40 °C. Subsequently, 20 mL of methanol was added to the system and stirred overnight at room temperature. The solvent was removed by distillation under reduced pressure to obtain 1.65 g of methyl 3,4,5-trimethoxyphenylacetic acid.

[0041] At room temperature, 1.5 g (6.25 mmol, 1.0 eq.) of methyl 3,4,5-trimethoxyphenylacetate was placed in a 100 mL Schlenk flask, along with 0.224 g (7.50 mmol, 1.2 eq.) of paraformaldehyde and 5 mL of DMSO. Under nitrogen protection, 0.0338 g (0.625 mmol, 10 mol%) of sodium methoxide was added as a catalyst, and the mixture was stirred overnight at room temperature. For post-treatment, the reaction mixture was transferred to 100 mL of water, and the aqueous layer was extracted with EtOAc. 20 mL of organic phase was combined, washed with water and dried, concentrated under reduced pressure and purified by column chromatography to obtain 1.17 g of methyl 3,4,5-trimethoxy-α-(hydroxymethyl)phenylacetate.

[0042] 1.0 g (3.70 mmol, 1.0 eq.) of methyl 3,4,5-trimethoxy-α-(hydroxymethyl)phenylacetate was placed in a 20 mL Schlenk flask. 2 mL of 40% hydrogen bromide solution was slowly added to the system at room temperature, and the mixture was heated under reflux overnight. For post-treatment, the reaction mixture was transferred to 50 mL of water, and the aqueous layer was extracted with DCM. 20 mL of organic phase was combined, washed with water and dried, concentrated under reduced pressure and purified by column chromatography to obtain 0.69 g of 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid.

[0043] 2. Structural Identification 3,4,5-Trihydroxy-α-(hydroxymethyl)phenylacetic acid is a white crystalline powder, readily soluble in methanol and soluble in water. 1 H-NMR (400 MHz, CD3OD) See Figure 3 , δ H (ppm) 3.63 (2H, m, CH2), 3.99 (1H, m, CH), 6.10(2H, s Ph-H); 13 C-NMR (400 MHz, CD3OD) See Figure 4 , δ C (ppm) 55.9 (CH), 65.2 (CH2),106.8 (2 CH), 131.0 (C), 131.3 (C), 146.2 (2 C), 176.2 (C=O).

[0044] Example 3: Study on the effects of tropinic acid and its derivatives on an immunosuppressed mouse model 1. Animal grouping, model establishment, and drug administration One hundred and ten male KM mice were randomly divided into 11 groups of ten mice each: a blank control group, a model group, a positive drug group, a high-dose troponic acid group (A), a low-dose troponic acid group (B), a high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), a low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), a high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), a low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), a high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and a low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (H). Except for the blank control group, which received no treatment, all other experimental mice were intraperitoneally injected with cyclophosphamide 80 mg / kg / day for 1–3 days at the start of the experiment to establish an immunosuppressed mouse model. All groups were administered the medication simultaneously, twice daily. See Table 1 for details of the grouping and administration regimens. The treatment lasted for 14 consecutive days.

[0045] Table 1. Animal grouping and drug administration information (n = 10) 2. Indicator Testing Mice were fasted for 12 hours after the last administration of the drug, then anesthetized by intraperitoneal injection of 10% chloral hydrate. They were weighed, dissected, and blood was drawn from the heart. Immediately afterward, the spleen and thymus were separated. The blood on the surface of the organs was blotted dry with filter paper, and then they were weighed. The spleen index and thymus index were calculated. Organ index = organ wet weight (g) / body weight (g) 100%. Mouse blood was collected in centrifuge tubes and centrifuged at 2800 r / min at low temperature (4℃) for 10 min. Serum was collected and stored at -20℃ for later use. The levels and activities of IL-4, IL-10, IFN-γ, TNF-α and IgG in serum were determined using an enzyme-linked immunosorbent assay (ELISA) kit.

[0046] 3. Experimental Results and Discussion Spleen and thymus indices can directly reflect the strength of the body's immune function. Compared with the blank control group, the thymus and spleen indices in the model group were significantly decreased. P <0.01%, after two weeks of treatment with troponic acid and its derivatives, the spleen and thymus indices in all dose groups increased significantly ( P <0.05 or P <0.01), and in a dose-dependent manner, the high-dose group increased the spleen and thymus indices and then tended to normalize. This indicates that tropic acid and its derivatives can significantly improve the immune organ indices of cyclophosphamide-induced immunosuppressed mice. IL-4 has immunomodulatory effects on B cells, T cells, mast cells, macrophages, and stem cells, and can induce the production of IgG and IgE. Compared with the model group, the serum IL-4 levels of mice in all dose groups of tropic acid and its derivatives were increased ( P<0.01), and the IL-4 level in the high-dose group increased and tended to normalize. IL-10 can inhibit NK cell activity and interfere with the production of cytokines by NK cells and macrophages. Compared with the model group, the serum IL-10 level in mice in the troponic acid and its derivative groups was significantly reduced ( P <0.05 or P <0.01%, the high-dose group reduced IL-4 levels and brought them closer to normal. Compared with the model group, the tropinic acid and its derivatives groups showed a significant increase in TNF-α levels ( P <0.05 or P <0.01), therefore, tropic acid and its derivatives can stimulate TNF-α secretion. TNF-α secretion induces enhanced macrophage activity and killing function, enabling macrophages to promote the body's immune response. Compared with the model group, the serum IgG concentration in mice in the tropic acid and its derivatives group was significantly increased ( P <0.05 or P <0.01), the high-dose group increased IgG levels and tended to normalize. There was no significant difference in therapeutic effect between compound 1 and compound 2. P >0.05%. Compared with compounds 1 and 2, compounds 3 and 4 showed better therapeutic effects ( P <0.05 or P <0.01). The effects of tropic acid and its derivatives on organ indices and serum immune parameters in an immunosuppressed mouse model are shown in Table 2. In summary, tropic acid and its derivatives have a certain regulatory effect on cyclophosphamide-induced immunosuppression.

[0047] Table 2. Effects of tropinic acid and its derivatives on organ indices and serum immune markers in an immunosuppressed mouse model. Note: Compared with the blank control group, ## P <0.01; compared with the model group, P <0.05, P <0.01.

[0048] Example 4: Study on the effects of tropinic acid and its derivatives on the proliferation of splenic lymphocytes 1. Cell model preparation, grouping, and drug administration Mice were euthanized by cervical dislocation, sterilized with 75% ethanol, and their spleens were removed under aseptic conditions. Splenic lymphocytes were prepared, resuspended in RPMI-1640 medium, and the cell concentration was adjusted to 1:1 by counting. 10 7Cells / ml. Add 100 μL of cell suspension to a 96-well plate. Add mitogen ConA to each well to a final concentration of 5 µg / ml. Add 100 μL of tropinic acid or its derivatives to each well, including high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (H). Add 100 μL of RPMI-1640 culture medium to the blank control group and 100 μL of RPMI-1640 culture medium containing cyclophosphamide to the positive control group. Each group has 3 replicates. See Table 3 for grouping and drug administration information. 37 ℃, 5% After culturing in CO2 for 44 hours, add 10 μL of 5 mg / mL MTT to each well and continue culturing for another 4 hours.

[0049] Table 3. Cell grouping and drug administration information (n = 9) 2. Indicator Testing After cell culture, the cells were centrifuged at 1000 r / min, the supernatant was discarded, and 200 μL of DMSO was added to each well. The cells were then shaken for 10 min to dissolve the cells. The cells were then placed in a microplate reader, and the OD value was measured at a wavelength of 492 nm. The experiment was performed in triplicate. Spleen lymphocyte proliferation inhibition rate = (mean OD value of ConA model control group - mean OD value of drug treatment group) / mean OD value of ConA model control group.

[0050] 3. Experimental Results and Discussion As shown in Table 4, none of the tropinic acid and its derivative groups showed cytotoxicity, and all exhibited varying degrees of immunosuppressive effects on ConA-induced spleen cell proliferation. Compared with the model group, the OD values ​​of each tropinic acid and its derivative groups were significantly reduced. P <0.01%, and the inhibition rate in the high-dose group reached 39.8%–44.5%. Compound 1 showed no significant difference in effect compared to compound 2. P >0.05). Compared with compounds 1 and 2, compounds 3 and 4 showed more significant effects ( P <0.01).

[0051] Table 4. Effects of tropinic acid and its derivatives on the proliferation of mouse spleen lymphocytes Note: Compared with the model group, P <0.01.

[0052] Example 5: Study on the effects of tropinic acid and its derivatives on delayed hypersensitivity reactions 1. Animal grouping, model establishment, and drug administration One hundred and ten male KM mice (6-7 weeks old, 22.0 ± 2.0 g) were randomly divided into 11 groups of 10 mice each: blank control group, model group, positive drug group, high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (H). On day 1 of drug administration, approximately 1.0 cm of hair was removed from the abdomen of each group of mice. Apply 50 μL of 1% 2,4-dinitrofluorobenzene (DNFB) (dissolved in acetone-olive oil, acetone:olive oil = 3:1) to a 2.0 cm area. For the normal control group, apply 50 μL of acetone-olive oil (3:1) to the abdomen. Administration was performed on the day of sensitization, twice daily for 7 consecutive days, with a second application on the second day for enhanced sensitization. All groups were administered medication simultaneously. See Table 5 for grouping and administration details.

[0053] Table 5. Animal grouping and drug administration information (n = 10) 2. Indicator Testing Thirty minutes after the last administration, 10 μL of 1% DNFB was applied to both the anterior and posterior surfaces of the right ear of each mouse. Twenty-four hours later, the mice were weighed, anesthetized by intraperitoneal injection of 10% chloral hydrate (0.1 mL / 10g), and euthanized by cervical dislocation after blood collection from enucleation. Both ears were removed, and circular ear pieces were punched at the same location using an 8 mm punch and accurately weighed. The blood was centrifuged, and the supernatant was collected. Serum IFN-γ levels were measured using an enzyme-linked immunosorbent assay (ELISA) kit. γ The content of [unclear]. Ear swelling degree = Right ear mass - Left ear mass; Swelling inhibition rate = (Average swelling degree of model group - Average swelling degree of treatment group) / Average swelling degree of model group 100%.

[0054] 3. Experimental Results and Discussion Delayed-type hypersensitivity (DHT) is a T-cell-dependent immune response model, a Th1-mediated allergic reaction. Th1 cells primarily secrete INF-γ, participating in cellular immunity and the occurrence of DHT inflammation. Its main characteristic is the appearance of a delayed-type inflammatory response at the antigen attack site in the sensitized organism. DNCB is a hapten; when diluted and applied to the abdominal skin, it binds to skin proteins to form a complete antigen, thereby stimulating T lymphocytes to proliferate into sensitized lymphocytes. Seven days later, it is applied to the ear to induce a local DHT reaction. As shown in Table 6, compared with the model group, tropic acid and its derivatives significantly reduced ear swelling in mice with DHT. P <0.05 or P <0.01), and significantly inhibited serum INF-1 in delayed-type hypersensitivity mice. γ Increase in content ( P <0.05 or P <0.01), showing a dose-dependent effect; high-dose groups could reduce INF levels in mouse ears and serum. γ The situation is trending towards normal. There is no significant difference in effect between compounds 1 and 2. P >0.05%. Compounds 3 and 4 showed better performance compared to compounds 1 and 2. P <0.05). This indicates that tropinic acid and its derivatives have a regulatory effect on delayed-type hypersensitivity reactions and inhibit serum INF-12-. γ Increased content may be its mechanism of action.

[0055] Table 6. Effects of tropinic acid and its derivatives on delayed hypersensitivity reactions in mice. Note: Comparison with the blank control group. ## P <0.01; compared with the model group, P <0.05, P <0.01.

[0056] Example 6: Study on the anti-inflammatory effects of tropinic acid and its derivatives 1. Animal grouping, model establishment, and drug administration One hundred and ten KM mice (6-7 weeks old, 22.0±2.0 g), half male and half female, were randomly divided into 11 groups of 10 mice each: blank control group, model group, positive drug group, high-dose troponic acid group (A), low-dose troponic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (H). All groups were administered the drug simultaneously. The grouping and administration regimens are detailed in Table 7. The drugs were administered twice daily by gavage for 7 consecutive days. The last administration was 1... After h, except for the normal control group, mice in the other groups had 40 μL of xylene evenly applied to both sides of the right ear to induce inflammation.

[0057] Table 7. Animal grouping and drug administration information (n = 10) 2. Indicator Testing One hour after inducing inflammation, the animals were anesthetized by intraperitoneal injection of 10% chloral hydrate, euthanized by cervical dislocation, and both ears were cut off along the auricle edge. Circular ear pieces were then made at the same location in both ears using a manual 8 mm ear punch. The ear pieces were precisely weighed, and the degree of ear swelling and the swelling inhibition rate were calculated. Ear swelling = Right ear mass - Left ear mass; Swelling inhibition rate = (Average swelling of the model group - Average swelling of the treatment group) / Average swelling of the model group 100%.

[0058] 3. Experimental Results and Discussion As shown in Table 8, compared with the model control group, tropinic acid and its derivative groups significantly reduced the degree of xylene-induced ear swelling in mice. P <0.01), with the high-dose groups of tropinic acid and its derivatives showing swelling inhibition rates of 70.34%–74.31%. There was no significant difference in swelling inhibition effect between compounds 1 and 2. Compared with compounds 1 and 2, compounds 3 and 4 showed better swelling inhibition effects in both high- and low-dose groups. In summary, these results indicate that tropinic acid and its derivatives have significant anti-inflammatory effects on the xylene-induced mouse ear swelling model.

[0059] Table 8. Effects of tropinic acid and its derivatives on xylene-induced ear swelling in mice Note: Compared with the blank control group, ## P <0.01; compared with the model group, P <0.01.

[0060] Example 7: Study on the analgesic effect of tropinic acid and its derivatives 1. Animal grouping, model establishment, and drug administration One hundred KM mice (5-6 weeks old, 22.0±2.0 g), half male and half female, were randomly divided into 10 groups of 10 mice each: blank control group, model group, positive drug group, high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (H). All groups were administered the drug simultaneously, twice daily by gavage for 7 consecutive days. The grouping and administration regimens are detailed in Table 9. Except for the normal control group, the mice in the other groups received an intraperitoneal injection of 0.6% tropinic acid 2 hours after the last administration. A pain model was established using 10 mL / kg glacial acetic acid.

[0061] Table 9. Animal grouping and drug administration information (n = 10) 2. Indicator Testing After establishing the pain model, the latency time for the writhing response in mice and the number of writhing episodes in each group within 20 minutes were observed and recorded, and the analgesia rate was calculated. Analgesia rate = (Average number of writhing episodes in the model group - Average number of writhing episodes in the treatment group) / Average number of writhing episodes in the model control group 100%.

[0062] 3. Experimental Results and Discussion As shown in Table 10, compared with the model control group, tropinic acid and its derivative groups significantly prolonged the latency of acetic acid-induced writhing response in mice. P <0.05 or P <0.01), and significantly reduced the number of twisting movements ( P <0.01), among which the high-dose group of tropinic acid and its derivatives achieved an analgesic rate of 55.26–58.79%. The analgesic effect of compound 1 was not significantly different from that of compound 2. Compared with compounds 1 and 2, compounds 3 and 4 showed better analgesic effects. Therefore, tropinic acid and its derivatives have significant peripheral analgesic effects.

[0063] Table 10. Effects of tropinic acid and its derivatives on acetic acid-induced writhing response in mice Note: Compared with the model group, P <0.05, P <0.01.

[0064] Example 8: Study on the effects of tropinic acid and its derivatives on a mouse model of autoimmune hepatitis 1. Animal modeling, grouping, and drug administration Liver tissue was taken from normal male KM mice (6-7 weeks old, 22.0±2.0 g), and physiological saline was added at a ratio of 1:9 (g / mL). The mixture was thoroughly homogenized using a tissue homogenizer (-4℃), centrifuged for 10 minutes (2500 rpm, 4℃), and the supernatant was collected to obtain the syngeneic liver antigen. The syngeneic liver antigen was added to complete Freund's adjuvant at a ratio of 1:1 (v / v), mixed thoroughly, and emulsified to obtain the immunomodulator (prepared immediately before use).

[0065] One hundred and ten male KM mice (6-7 weeks old, 22±2g) were randomly divided into 11 groups of 10 mice each: blank control group, model group, positive drug group, high-dose troponic acid group (A), low-dose troponic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Except for the blank control group, all mice in each group received an intraperitoneal injection of 1 mL of the immunizing agent for the first immunization. A second immunization was performed 7 days later to obtain a mouse model of autoimmune hepatitis. The blank control group mice were injected intraperitoneally with 1 mL of physiological saline per mouse at the same time. The drugs were administered by gavage starting on the day the model was established, twice a day for 14 consecutive days. The grouping and administration regimens are detailed in Table 11.

[0066] Table 11. Animal grouping and drug administration information (n = 10) 2. Indicator Testing After the last administration, mice were fasted for 12 hours. Mice in each group were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.1 mL / 10g), then dissected. Blood was collected from the heart, perfused with physiological saline, and fixed with paraformaldehyde. Liver and spleen tissues were collected and fixed in 4% paraformaldehyde for 24 hours. Blood was centrifuged for 10 minutes (3000 rpm, 4℃), and serum was collected. The activities of liver function-related indicators such as lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), as well as the content of total bilirubin (TBIL), were measured according to the kit instructions. Liver and spleen tissues were paraffin-embedded, cut into 4 μm thick sections, routinely stained with hematoxylin and eosin (HE), mounted, and observed under a light microscope for histopathological changes.

[0067] 3. Experimental Results and Discussion Liver function tests: As shown in Table 12, the activities of LDH, ALT, and AST in the serum of mice in the model group were increased, and the content of TBIL was significantly increased. P <0.01). When liver tissue is damaged or hepatocytes die, the activities of LDH, ALT, and AST in serum increase, while the level of TBIL increases. This is a sensitive indicator of liver injury, directly reflecting the degree of hepatocyte damage and necrosis, as well as the liver's detoxification and metabolic functions. Compared with the model group, after 2 weeks of administration of troponic acid and its derivatives, the activities of LDH, ALT, AST, and the level of TBIL in serum all decreased. P <0.05 or P <0.01), and the effect was dose-dependent. There was no significant difference in therapeutic effect between compound 1 and compound 2 ( P >0.05%. Compared with compounds 1 and 2, compounds 3 and 4 showed better therapeutic effects ( P <0.05). Therefore, tropinic acid and its derivatives can improve liver function in mice with an autoimmune hepatitis model.

[0068] Table 12. Effects of tropinic acid and its derivatives on liver function in a mouse model of autoimmune hepatitis Note: Compared with the blank control group, ## P <0.01; compared with the model group, P <0.05, P <0.01.

[0069] Histopathological examination: such as Figure 5As shown, the livers of model group mice exhibited extensive infiltration of inflammatory factors, blurred cell boundaries, hepatocyte edema and degeneration, nuclear pyknosis, partial cellular lysis and necrosis, and vacuolation caused by fatty degeneration. After administration of tropinic acid and its derivatives, some of the above pathological indicators showed significant improvement. In particular, regenerated hepatocytes repairing liver damage were observed around necrotic hepatocytes, characterized by large cell volume, large and deeply stained nuclei, and mostly binucleated. Figure 6 As shown, in the spleen of model mice, follicular proliferative lesions related to immune activation appeared, leading to an increase and enlargement of germinal centers. The ratio of red pulp to white pulp area decreased, and the number of foamy macrophages in both the white and red pulp increased. This is attributed to a large-scale lymphocyte transformation in the spleen following intraperitoneal injection of immunizing agents, with B cells proliferating and transforming into plasma cells. The expansion of splenic follicular growth centers and the presence of numerous plasma cells increased antibody production and enhanced humoral immunity. Plasma was present in the blood vessels of the red pulp, and the proliferating macrophages destroyed the plasma and erythrocytes in the red pulp arterioles, resulting in massive congestion of the splenic cords within the red pulp. After administration of tropic acid and its derivatives, the above-mentioned immune activation pathological indicators significantly improved. This suggests that tropic acid and its derivatives may regulate immune activity by enhancing antigen recognition, thereby playing a therapeutic role in autoimmune hepatitis.

[0070] Example 9: Study on the effects of tropinic acid and its derivatives on a mouse model of pulmonary inflammation caused by novel coronavirus and cold-dampness epidemic 1. Animal modeling, grouping, and drug administration One hundred and ten male KM mice (6-7 weeks old, 22.0±2.0 mm) were randomly divided into 11 groups of 10 mice each: blank control group, model group, positive drug group, high-dose troponic acid group (A), low-dose troponic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Each group was administered the drug twice daily for seven consecutive days. Three days after administration, except for the control group, all other groups of mice were intraperitoneally injected with 5 mg / kg lipopolysaccharide saline solution. They were then placed in an artificial climate chamber at a controlled temperature of 4.0±2.0℃ and humidity of 90.0±3.0% to induce a cold-dampness stimulation model, which was maintained for 8 hours daily for 4 consecutive days, thus creating a mouse model of lung inflammation caused by the novel coronavirus. See Table 13 for grouping and administration details.

[0071] Table 13. Animal grouping and drug administration information (n = 10) 2. Indicator Testing After the last administration, the mice were fasted for 12 hours, anesthetized by intraperitoneal injection of 10% chloral hydrate (0.1 mL / 10g), perfused with physiological saline, and lung tissue was harvested. A portion was flash-frozen in liquid nitrogen and stored at -80℃ for later use, while another portion was fixed in 4% paraformaldehyde for later use. 50 mg of the frozen mouse tissue was homogenized with 800 μL PBS, centrifuged at 2800 r / min at 4℃ for 10 min, and serum was collected. The levels of inflammatory factors IL-6, IL-10, IFN-γ, and TNF-α in the lung tissue were measured using an enzyme-linked immunosorbent assay (ELISA) kit. The fixed lung tissue was embedded in paraffin, sectioned into 4 μm thick sections, routinely stained with hematoxylin and eosin (HE), mounted, and observed under a light microscope for histopathological changes.

[0072] 3. Experimental Results and Discussion Inflammatory factor detection: As shown in Table 14, compared with the blank control group, the levels of TNF-α, IFN-γ, and IL-6 in the lung tissue of the model group mice were significantly increased ( P <0.05 or P <0.01), IL-10 levels were significantly reduced ( P <0.01%, after 7 days of treatment with tropinic acid and its derivatives, the levels of TNF-α, IFN-γ, and IL-6 all decreased significantly ( P <0.05 or P <0.01), IL-10 levels were significantly elevated ( P <0.05 or P <0.01), and the effect was dose-dependent; the high-dose group reduced lung tissue inflammatory factors and brought them closer to normal. There was no significant difference in therapeutic effect between compound 1 and compound 2. P >0.05%. Compared with compounds 1 and 2, compounds 3 and 4 showed better therapeutic effects ( P <0.05). Therefore, tropic acid and its derivatives can improve the inflammatory response in a mouse model of lung inflammation caused by novel coronavirus.

[0073] Table 14. Effects of tropinic acid and its derivatives on inflammatory factors in lung tissue of a mouse model of pulmonary inflammation induced by novel coronavirus and cold-dampness epidemic. Note: Compared with the blank control group, ## P <0.01; compared with the model group, P <0.05, P <0.01.

[0074] Histopathological examination: such as Figure 7 As shown, the lung tissue structure of the normal group mice was intact, without exudate, and the alveolar septa were normal; while the alveolar scaffold of the model group mice collapsed, the lung tissue structure was disordered, the alveolar septa were significantly thickened, and the interstitial edema and inflammatory infiltration of the lung tissue were severe. After administration of tropic acid and its derivatives, some of the above pathological indicators were significantly improved. There was no significant difference in the therapeutic effect between compounds 1 and 2. Compared with compounds 1 and 2, compounds 3 and 4 had better therapeutic effects. This indicates that tropic acid and its derivatives may exert a therapeutic effect on the cold-dampness epidemic mouse model of lung inflammation caused by novel coronavirus by regulating immunity and inhibiting the inflammatory response.

[0075] Example 10: Study on the effects of tropinic acid and its derivatives on a mouse model of aplastic anemia 1. Animal modeling, grouping, and drug administration Sixty-six male SD rats (7-8 weeks old, 200.0±20.0g) were randomly divided into 11 groups of 6 rats each: blank control group, model group, positive drug group, high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Except for the blank control group, the other groups were subcutaneously injected with 2% acetylphenylhydrazine (Ap) saline solution 20 mg / kg on day 1 and day 4, and acetylphenylhydrazine was injected with 2% acetylphenylhydrazine solution 20 mg / kg on day 4. A pneumothelial anemia animal model was established by intraperitoneal injection of cyclophosphamide (Cy) saline solution 20 mg / kg on days 5-7. All rats in each treatment group received the drug via tail vein injection once daily for 14 consecutive days. See Table 15 for grouping and administration details.

[0076] Table 15. Animal grouping and drug administration information (n = 6) 2. Indicator Testing One hour after the last administration of medication, rats were anesthetized via intraperitoneal injection of 10% chloral hydrate. Blood was collected from the heart in anticoagulant tubes for later use. The thymus and spleen were weighed. A hemolytic agent was added to the whole blood in the anticoagulant tubes, and a fully automated hematology analyzer was used to detect changes in peripheral blood cell counts, primarily including changes in four indicators: white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), and platelet count (PLT). The spleen index and thymus index were calculated: organ index = organ wet weight (g) / body weight (g). 100%.

[0077] 3. Experimental Results and Discussion As shown in Table 16, compared with the normal group, the levels of WBC, RBC, HGB, and PLT in the model group were significantly decreased. P <0.01); Compared with the model group, after 14 days of treatment with troponic acid and its derivatives, the levels of all indicators increased ( P <0.05 or P <0.01), and showed a dose-dependent effect. Compared with the normal group, the spleen index was significantly increased and the thymus index was significantly decreased in the model group ( P <0.01), after 14 days of treatment with troponic acid and its derivatives, the spleen index and thymus index tended to normalize. P <0.05 or P <0.01). There was no significant difference in therapeutic effect between compound 1 and compound 2. P >0.05%. Compounds 3 and 4 showed better therapeutic effects compared to compounds 1 and 2. P <0.05 or P <0.01). Therefore, tropinic acid and its derivatives can improve aplastic anemia in rat models through their immunomodulatory effects, thereby restoring the function of their hematopoietic stem cells.

[0078] Table 16. Effects of tropinic acid and its derivatives on immune organ indices and peripheral blood counts in an aplastic anemia model. Note: Compared with the blank control group, ## P <0.01; compared with the model control group, P <0.05, P <0.01.

[0079] Example 11: Study on the effects of tropinic acid and its derivatives on a rat model of rheumatoid arthritis 1. Animal modeling, grouping, and drug administration Preparation of immunoemulsifiers: 7 ml of bovine type II collagen (CII) solution was placed in a small beaker and magnetically stirred at 1500 rpm at low temperature. 7 ml of complete Freund's adjuvant (CFA) solution was slowly added to the CII solution. After all the CFA solution was added, stirring continued for approximately 30 minutes until the emulsion did not disperse when dropped onto water, thus obtaining the primary immunoemulsifier. Secondary immunoemulsifiers were obtained by replacing CFA with incomplete Freund's adjuvant (IFA) using the same preparation method. All immunoemulsifiers were prepared immediately before use.

[0080] Preparation of the rheumatoid arthritis (CIA) model: One hundred male SD rats (7-8 weeks old, 200.0±20.0g) were used. 0.2ml of the initial immunization emulsion was injected subcutaneously at the base of the tail. Eight days after the initial immunization, 0.1ml of the secondary immunization emulsion was injected subcutaneously at the base of the tail to complete the booster immunization. The blank control group received saline injection using the same method. Fourteen days after the booster immunization, the Arthritis Index (AI) was assessed using the following scoring rules: 0 points for no swelling or erythema; 1 point for erythema and mild swelling at the ankle joint; 2 points for erythema and mild swelling at the ankle to metatarsophalangeal or metacarpophalangeal joints; 3 points for erythema and moderate swelling at the ankle to metatarsophalangeal or metacarpophalangeal joints; and 4 points for erythema and severe swelling at the ankle to toe joints. The sum of the scores for both paws was used as the joint score for each rat. An AI score ≥4 indicated successful model establishment, and animals without signs of joint swelling were removed.

[0081] Grouping and administration: Six blank control rats and 60 CIA model rats were randomly divided into 10 groups of 6 rats each: model group, positive drug group, high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Each group was simultaneously administered the corresponding drug by gavage twice daily for 4 consecutive weeks. The grouping and administration regimens are detailed in Table 17.

[0082] Table 17. Animal grouping and drug administration information (n = 6) 2. Indicator Testing After treatment, the morphological differences in the ankle joint and paw of rats before and after treatment were compared among the groups. The arthritis status of rats in each group was scored weekly using the AI ​​scoring method described above. The volume change below the ankle joint was simultaneously measured using the paw volume displacement method, and the degree of joint swelling was calculated. Joint swelling degree = (volume after modeling - volume before modeling) / volume before modeling 100%.

[0083] 3. Experimental Results and Discussion like Figure 8 As shown, after modeling, rats exhibited a significant increase in metatarsal volume. In all model groups, rats showed joint swelling, stiff and curled paws, and were unable to walk normally. After 4 weeks of treatment with tropic acid and its derivatives, the low-dose group showed mild joint swelling, while the swelling subsided and the rats in the other treatment groups were in good condition. Compounds 1 and 2 showed no significant difference in therapeutic effect. Compared to compounds 1 and 2, compounds 3 and 4 showed better therapeutic effects. Figure 9 As shown, compared with the model group, the AI ​​score of the tropinic acid and its derivative treatment group gradually decreased, and the difference became statistically significant starting from the 3rd week of treatment. P <0.05 or P <0.01); Compared with the model group, after 2 weeks of treatment with tropinic acid and its derivatives, the joint swelling of rats began to decrease, and after 4 weeks of treatment, the joint swelling of rats decreased significantly. There was no significant difference in the treatment effect of compound 1 compared with compound 2 at each week (…). P >0.05%. Compared with compounds 1 and 2, after 3 weeks of administration, the high-dose groups of compounds 3 and 4 showed a more significant reduction in AI scores and joint swelling in rats. P (<0.05), indicating better therapeutic effect. In summary, tropinic acid and its derivatives have significant therapeutic effects on a rat model of rheumatoid arthritis.

[0084] Example 11: Study on the effects of tropinic acid and its derivatives on a mouse model of allergic rhinitis 1. Animal modeling, grouping, and drug administration One hundred and ten male BABL / c mice (5-6 weeks old, 20.0±2.0g) were randomly divided into 11 groups of 10 mice each: blank control group, model group, positive drug group, high-dose troponic acid group (A), low-dose troponic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Except for the blank control group, mice in the other groups were intraperitoneally injected with a suspension of sensitizing agents (containing 0.50 mg / mL OVA and 0.50 mg / mL OVA) on days 1, 7, and 14 of the experiment. Al(OH)3) was administered to each mouse at a dose of 200 μL for basal sensitization. From days 15 to 28, each group was administered the corresponding drug via gavage. From days 22 to 28, a 4% OVA solution was instilled into the nasal cavity of each mouse for challenge, with 20 μL in each nasal cavity, thus establishing an allergic rhinitis model. The blank control group received the same volume of physiological saline during both the basal sensitization and challenge phases. See Table 18 for grouping and drug administration details.

[0085] Table 18. Animal grouping and drug administration information (n = 10) 2. Indicator Testing After the last nasal stimulation, the number of times mice in each group scratched their noses and sneezed within 30 minutes was observed and recorded.

[0086] 3. Experimental Results and Discussion As shown in Table 19, compared with the blank group, the number of sneezes and nose scratches in the model group mice was significantly increased. P <0.01 indicates that the allergic rhinitis model was successfully established. Compared with the model group, tropinic acid and its derivatives significantly reduced the number of sneezes in mice. P <0.01) and the number of times the nose was scratched ( P <0.05 or P <0.01), and the effect was dose-dependent. There was no significant difference in therapeutic effect between compound 1 and compound 2 ( P >0.05%. Compounds 3 and 4 showed better therapeutic effects compared to compounds 1 and 2. P <0.05). This indicates that tropinic acid and its derivatives have a significant therapeutic effect on a mouse model of allergic rhinitis.

[0087] Table 19. Effects of tropinic acid and its derivatives on the number of nose scratches and sneezes in a mouse model of allergic rhinitis. Note: Compared with the blank control group, ## P <0.01; compared with the model control group, P <0.05, P <0.01.

[0088] Example 12: Study on the effects of tropinic acid and its derivatives on a rat model of membranous nephritis 1. Animal modeling, grouping, and drug administration Replication of the membranous glomerulonephritis model: 100 mg of cationized bovine serum albumin (C-BSA) was dissolved in 15 mL of physiological saline and mixed with an equal volume of incomplete Freund's adjuvant, then thoroughly emulsified. One hundred male SD rats (7–8 weeks old, 200±20 g) were used. 1 mL of the emulsifier was injected subcutaneously at multiple points in the bilateral axillae and groin of each rat, once every other day for a total of 3 times, to complete the pre-immunization. One week after pre-immunization, each rat was injected intravenously with C-BSA physiological saline solution at a dose of 16 mg / kg, 3 times a week for 4 consecutive weeks, to complete the formal immunization. Urine was collected from rats using metabolic cages for 24-hour urine protein testing. A 24-hour urine protein (24h UPro) level >20 mg was considered a successful replication of the membranous glomerulonephritis model, and rats with failed model replication were removed. The blank control group rats were injected with an equal volume of physiological saline simultaneously using the same method.

[0089] Grouping and administration: Six rats were used as the blank control group and 60 rats were used as the membranous nephritis model. The membranous nephritis model rats were randomly divided into 10 groups of 6 rats each. The groups were: model group, positive drug group, high-dose tropinic acid group (A), low-dose tropinic acid group (B), high-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (C), low-dose 4-hydroxy-α-(hydroxymethyl)phenylacetic acid group (D), high-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (E), low-dose 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid group (F), high-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G), and low-dose 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid group (G). Each group was simultaneously administered the corresponding drug by gavage twice a day for 4 consecutive weeks. The grouping and administration regimens are detailed in Table 11.

[0090] Table 20. Animal grouping and drug administration information (n = 6) 2. Indicator Testing General observation: After the last administration, the therapeutic effect of the drug was evaluated by comparing and observing the hair, mental state, weight, food intake, and stool and urination of rats in each group.

[0091] Quantitative detection of urinary protein and detection of serum biochemistry: 24-hour urine of rats was collected and the concentration of urinary protein was measured using a rat ELISA kit; rats were anesthetized and serum was collected, and the levels of serum total protein (TP), serum albumin (Alb), serum creatinine (Scr), and serum blood urea nitrogen (BUN) were measured using a fully automated biochemical analyzer.

[0092] 3. Experimental Results and Discussion General observation: In the normal group, rats showed normal activity, quick reflexes, glossy fur, normal appetite, and increased body weight. In the model group, rats exhibited poor mental state and hair luster, hair loss, decreased food intake, increased urine output, loose stools, and subcutaneous edema in the abdomen of some rats. The tropinic acid and its derivatives groups showed significant improvements in mental state, fur, appetite, and bowel movements.

[0093] Quantitative detection of urinary protein and serum biochemistry: As shown in Table 21, compared with the normal group, the model group rats showed significantly elevated proteinuria, serum Scr and BUN (… P <0.01%, serum TP and Alb levels were significantly decreased ( P <0.01); After 4 weeks of treatment with troponic acid and its derivatives, the 24-hour urinary protein level in rats was significantly reduced ( P <0.01), serum TP and Alb levels were significantly elevated ( P <0.05 or P <0.01, and tended to normal; compared with the normal group, the serum Scr and BUN levels in the model group were significantly increased ( P <0.01%, after 4 weeks of treatment with troponic acid and its derivatives, serum Scr and BUN levels decreased to varying degrees. P <0.05 or P <0.01), and tended to normal. There was no significant difference in therapeutic effect between compound 1 and compound 2 ( P >0.05%. Compounds 3 and 4 showed better therapeutic effects compared to compounds 1 and 2. P <0.05). Therefore, tropinic acid and its derivatives have the effect of improving or restoring renal function in rat models of membranous nephritis.

[0094] Table 21. Effects of tropinic acid and its derivatives on renal function in a rat model of membranous nephritis Note: Compared with the blank control group, ## P <0.01; compared with the model group, P <0.05, P <0.01.

[0095] Example 13: Preparation of tropinic acid and its derivative tablets The prescriptions are shown in Table 2. According to prescription 1, take the active pharmaceutical ingredient (tropic acid) and excipients (lactose, microcrystalline cellulose, polyvinylpyrrolidone, croscarmellose sodium, microcrystalline silica, magnesium stearate, and purified water). Add tropic acid, lactose, microcrystalline cellulose, and polyvinylpyrrolidone to a wet granulator, using purified water as a wetting agent for wet granulation. After wet granulation, drying, and dry granulation, dry granules are obtained. Add croscarmellose sodium, microcrystalline silica, and magnesium stearate to the dry granules and mix thoroughly. Then, compress the thoroughly mixed material into tablets using a tableting machine to obtain tropic acid tablets. Take the active pharmaceutical ingredient and excipients from prescriptions 2, 3, and 4 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid tablets, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid tablets, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid tablets using the same process as above.

[0096] Table 22. Formulation of troponic acid and its derivatives tablets Example 14: Preparation of tropinic acid and its derivatives coated tablets The prescription is shown in Table 3. Take the active pharmaceutical ingredient (tropic acid) and excipients (lactose, microcrystalline cellulose, hydroxypropyl methylcellulose, sodium lauryl sulfate, sodium carboxymethyl starch, micronized silica gel, magnesium stearate, film coating premix, and purified water) according to prescription 5. Add sodium lauryl sulfate to purified water and stir until dissolved. Add tropic acid, lactose, microcrystalline cellulose, and hydroxypropyl methylcellulose to a wet granulator. Use an aqueous solution of sodium lauryl sulfate as a wetting agent for wet granulation. After wet granulation, drying, and dry granulation, dry granules are obtained. Add sodium carboxymethyl starch, micronized silica gel, and magnesium stearate to the dry granules and mix thoroughly. Compress the thoroughly mixed material into tablets using a tableting machine to obtain uncoated tropic acid tablets. Add the film coating premix to purified water and stir continuously for more than 1 hour to obtain a film coating solution. Then, coat the obtained uncoated tablets in a coating machine to obtain tropic acid coated tablets. Take the active pharmaceutical ingredients and excipients from prescriptions 6, 7, and 8 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid coated tablets, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid coated tablets, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid coated tablets respectively using the same process as described above.

[0097] Table 23. Formulations for tropinic acid and its derivatives coated tablets Example 15: Preparation of Tropical Acid and its Derivatives Spray I The prescription is shown in Table 4. According to prescription 9, take the active pharmaceutical ingredient (tropic acid) and excipients (polyvinylpyrrolidone K30, propylene glycol monooctanoate, ethylparaben, poloxamer, di-tert-butyl-p-cresol (BHT), 1N sodium hydroxide solution, ethanol, and water). Add the active pharmaceutical ingredient, polyvinylpyrrolidone K30, propylene glycol monooctanoate, ethylparaben, and BHT to ethanol and stir until completely dissolved. Dissolve poloxamer in an appropriate amount of water. Then add the poloxamer aqueous solution to the ethanol solution of the above mixture, and adjust the pH to the range of 3-8 with 1N sodium hydroxide solution. Add water to a final volume of 200 ml to obtain the spray solution. Fill the solution into spray bottles to obtain tropic acid spray I. Take the active pharmaceutical ingredients and excipients of prescriptions 10, 11 and 12 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid spray I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid spray I and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid spray I respectively according to the same process method as above.

[0098] Table 24. Formulation of Tropine Acid and its Derivatives Spray I Example 16: Preparation of Tropical Acid and its Derivatives Spray II The prescriptions are shown in Table 5. According to prescription 13, take the active pharmaceutical ingredient (tropic acid) and excipients (hydroxypropyl cellulose, propylene glycol monooctanoate, ethylparaben, ethanol, and water). Add the active pharmaceutical ingredient, hydroxypropyl cellulose, propylene glycol monooctanoate, and ethylparaben to ethanol, stir until completely dissolved, and add water to a final volume of 200 ml to obtain the spray solution. Fill the solution into spray bottles to obtain tropic acid spray II. Take the active pharmaceutical ingredients and excipients from prescriptions 14, 15, and 16 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid spray II, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid spray II, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid spray II respectively using the same process as described above.

[0099] Table 25. Formulation of Tropine Acid and its Derivatives Spray II Example 17: Preparation of Tropical Acid and its Derivatives Injection I The prescriptions are shown in Table 6. According to prescription 17, take the active pharmaceutical ingredient (tropic acid) and excipients (sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, poloxamer, sodium bisulfite, and water for injection). Dissolve the active pharmaceutical ingredient and poloxamer in water for injection. Then add sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, and sodium bisulfite to the above solution and dissolve them. Finally, add water to 100 mL to obtain the injection solution. Fill the above injection solution into ampoules or vials of the appropriate volume to obtain tropic acid injection I. Take the active pharmaceutical ingredients and excipients from prescriptions 18, 19, and 20 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid injection I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid injection I, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid injection I respectively, following the same process as above.

[0100] Table 26. Prescriptions for Tropical Acid and its Derivatives Injectable Formulation I Example 18: Preparation of Tropical Acid and its Derivatives Injection II The prescriptions are shown in Table 7. According to prescription 21, take the active pharmaceutical ingredient (tropic acid) and excipients (sodium chloride, disodium hydrogen phosphate, sodium dihydrogen phosphate, Tween 80, and water for injection). Dissolve the active pharmaceutical ingredient and Tween 80 in water for injection. Then, add sodium chloride, disodium hydrogen phosphate, and sodium dihydrogen phosphate to the above solution and dissolve them. Finally, add water to 100 mL to obtain the injection solution. Fill the above injection solution into ampoules or vials of the appropriate volume to obtain tropic acid injection II. Take the active pharmaceutical ingredients and excipients from prescriptions 22, 23, and 24 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid injection II, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid injection II, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid injection II respectively using the same process as above.

[0101] Table 27. Formulation of Tropical Acid and its Derivatives Injection II Example 19: Preparation of Tropical Acid and its Derivatives Cream I The prescription is shown in Table 8. Take the active pharmaceutical ingredient (tropinic acid) and excipients (white petrolatum, cetyl alcohol, Tween 80, diethylene glycol monoethyl ether, ethylparaben, BHT, propylene glycol, citric acid / sodium citrate and water) according to prescription 25. The preparation process is as follows: (1) Dissolving the active pharmaceutical ingredient: Weigh propylene glycol, add the active pharmaceutical ingredient, and stir to dissolve at 40~50℃; (2) Preparing the oil phase: Weigh white petrolatum, cetyl alcohol, ethylparaben and BHT, heat to 60~80℃, stir to dissolve, and add the dissolved active pharmaceutical ingredient. (2) Add the raw materials slowly, continue stirring, mix evenly, and set aside; (3) Aqueous phase preparation: Weigh purified water, add Tween 80 and diethylene glycol monoethyl ether, heat to 60~80℃, adjust the pH value with citric acid / sodium citrate, stir to dissolve and set aside; (4) Emulsification: Slowly add the aqueous phase to the oil phase, keep at 70℃, homogenize, and continue stirring for more than 30 minutes; (5) Ointment formation: Cool down, stop heating, continue stirring, gradually cool to room temperature and cool to form an ointment, and fill into containers. Tropine acid cream I is obtained. Take the raw materials and excipients of prescriptions 26, 27 and 28 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid cream I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid cream I and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid cream I respectively according to the same process method as above.

[0102] Table 28. Formulation of Tropical Acid and its Derivatives Cream I Example 20: Preparation of Tropical Acid and its Derivatives Cream II The prescription is shown in Table 9. According to prescription 29, the active pharmaceutical ingredient (tropic acid) and excipients (white petrolatum, cetyl alcohol, poloxamer 407, propylene glycol monooctanoate, sodium benzoate, BHA, propylene glycol, glacial acetic acid / sodium acetate, and water) are prepared as follows: (1) Dissolving the active pharmaceutical ingredient: Weigh propylene glycol, add the active pharmaceutical ingredient, and stir to dissolve at 40-50℃; (2) Preparing the oil phase: Weigh white petrolatum, cetyl alcohol, propylene glycol monooctanoate, and BHA, heat to 60-80℃, stir to dissolve, and then... (2) Add the raw materials of the solution slowly, continue stirring, mix evenly, and set aside; (3) Aqueous phase preparation: Weigh purified water, add poloxamer 407 and sodium benzoate, heat to 60~80℃, adjust the pH value with glacial acetic acid / sodium acetate, stir to dissolve and set aside; (4) Emulsification: Slowly add the aqueous phase to the oil phase, keep at 70℃, homogenize, and continue stirring for more than 30 minutes; (5) Ointment formation: Cool down, stop heating, continue stirring, gradually cool to room temperature and cool to form an ointment, and fill into the container. Tropine cream II is obtained. Take the raw materials and excipients of prescriptions 30, 31 and 32 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid cream I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid cream I and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid cream II according to the same process method as above.

[0103] Table 29. Formulation of Tropine Acid and its Derivatives Cream II Example 21: Preparation of nasal formulation I of tropinic acid and its derivatives The prescriptions are shown in Table 30. According to prescription 33, take the active pharmaceutical ingredient (tropic acid) and excipients (sorbitol, citric acid / sodium citrate, sodium benzoate, and purified water). The preparation process is as follows: Dissolve sorbitol in purified water, then add the active pharmaceutical ingredient and sodium benzoate to the above solution and dissolve them. Adjust the pH to the range of 4.0-6.5 with citric acid / sodium citrate, and add purified water to 100 ml to obtain the nasal preparation solution. Finally, fill the solution into dropper bottles or spray bottles to obtain tropic acid nasal preparation I. Take the active pharmaceutical ingredients and excipients from prescriptions 34, 35, and 36 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid cream I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid cream I, and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid nasal preparation I respectively using the same process as above.

[0104] Table 30. Formulations of Tropical Acid and its Derivatives for Nasal Use (I) Example 22: Preparation of Tropical Acid and its Derivatives Nasal Formulation II The prescription is shown in Table 31. Take the active pharmaceutical ingredient (tropic acid) and excipients (sodium chloride, hydroxypropyl methylcellulose, Tween 80, sodium metabisulfite, sodium EDTA, 1N sodium hydroxide solution, benzalkonium chloride, and purified water) according to prescription 37. The preparation process is as follows: Add hydroxypropyl methylcellulose to purified water and stir continuously until completely dissolved. Add sodium chloride and Tween 80 to the hydroxypropyl methylcellulose solution and dissolve. Then add the active pharmaceutical ingredient, sodium metabisulfite, sodium EDTA, and benzalkonium chloride to the above solution and dissolve. Adjust the pH value to the range of 4.5-6.5 with 1N sodium hydroxide solution. Add purified water to 100 ml to obtain the nasal preparation solution. Finally, fill the solution into dropper bottles or spray bottles to obtain tropic acid nasal preparation II. Take the active pharmaceutical ingredients and excipients from prescriptions 38, 39 and 40 respectively, and prepare 4-hydroxy-α-(hydroxymethyl)phenylacetic acid cream I, 3,4-dihydroxy-α-(hydroxymethyl)phenylacetic acid cream I and 3,4,5-trihydroxy-α-(hydroxymethyl)phenylacetic acid nasal preparation II according to the same process as described above.

[0105] Table 31. Formulations for Tropical Acid and its Derivatives Nasal Preparations II The preferred embodiments of the present invention have been described above, but are not intended to limit the invention. Those skilled in the art can make modifications and variations to the embodiments disclosed herein without departing from the scope and spirit of the invention.

Claims

1. Use of tropinic acid and its derivatives, and pharmaceutically acceptable salts thereof, in the preparation of medicaments for the prevention and / or treatment of immune and inflammatory diseases, wherein the tropinic acid and its derivatives have the structure shown in Formula A: in, R1-R5 are each independently selected from -H or -OH; The immune and inflammation-related diseases mentioned are selected from: nephritis.

2. The use according to claim 1, characterized in that, The immune and inflammation-related diseases mentioned are selected from: membranous nephritis.

3. The use according to claim 1, characterized in that, The tropinic acid and its derivatives are selected from the compounds shown in Formulas I-IV below: 。 4. The use according to claim 1, characterized in that, The medication is a topical, oral, or injectable medication.

5. The use according to claim 1, characterized in that, The drug also contains pharmaceutically acceptable carriers or excipients.

6. The use according to claim 1, characterized in that, The drug is in solid, liquid, or semi-solid dosage form.

7. The use according to claim 1, characterized in that, The dosage forms of the medicine are: powder, tablet, granule, capsule, injection, spray, aerosol, powder mist, lotion, liniment, ointment, plaster, and patch.

8. The use according to claim 1, characterized in that, The dosage forms of the drug are: coated tablets, solutions, emulsions, suspensions, pastes, and gels.

9. The use according to claim 1, characterized in that, The dosage form of the drug is: nasal preparation.

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