Pentacyclic triterpene derivative as well as preparation method and application thereof

Abstract

The invention relates to the field of medicinal chemistry and medicine, and discloses pentacyclic triterpene derivatives and application thereof in prevention and / or treatment of immune diseases. The pentacyclic triterpene derivative disclosed by the invention has a structure as shown in a formula (I) and has an excellent immunoregulation effect on Tc1, Tc17 and Th17 cells, and the invention also provides application of the compound in preparation of medicines for preventing and / or treating immune diseases.

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry and pharmaceuticals; more specifically, it relates to the use of pentacyclic triterpenoid derivatives or pharmaceutically acceptable salts thereof in the prevention and / or treatment of immune diseases. Background Technology

[0002] T lymphocytes can be divided into several subsets, including helper T cells (CD4+), cytotoxic T cells (CD8+), and regulatory T cells (Tregs). These different subsets play different roles in the immune response. For example, helper T cells can stimulate B cells to produce antibodies, while cytotoxic T cells can directly kill virus-infected cells.

[0003] Helper T cell 17 (Th17) is a CD4+ cell. + Effector subsets of T cells secrete the pro-inflammatory cytokine IL-17, playing a crucial role in the development of various inflammatory diseases. They are particularly important in the occurrence and progression of certain inflammatory diseases, including autoimmune diseases. Increasing evidence shows a significant increase in the number of Th17 cells and related cytokines in the blood of patients with various autoimmune diseases, suggesting that targeting Th17 cell differentiation may become a novel clinical treatment option for these diseases. Biologics targeting the IL-17 / Th17 pathway have shown positive effects, with secukinumab, ixekizumab, and piracetamab among those approved for marketing.

[0004] According to CD8 + T cells secrete cytokines and have different functions, and can be classified into different cytotoxic T lymphocyte (Tc cell) subtypes, including Tc1, Tc2, Tc17, and Tc22 cells. Tc1 cells can secrete IFN-γ, granzymes, and perforin, thereby killing melanocytes and leading to the development and progression of vitiligo. Tc17 cells mainly exert their biological effects by secreting inflammatory factors, and multiple studies have reported their important role in the pathogenesis of autoimmune diseases such as psoriatic arthritis and type 1 diabetes.

[0005] Pentacyclic triterpenoids extracted from medicinal plants, fruits, and vegetables, such as ursolic acid, oleanolic acid, glycyrrhetinic acid, and boswellic acid, possess a wide range of biological activities, including antitumor, anti-inflammatory, hepatoprotective, antiviral, antidiabetic, and antibacterial effects. However, they suffer from drawbacks such as low activity, poor water solubility, and low bioavailability. Therefore, modifying their compound structures to discover superior candidate drugs has significant scientific research value and social benefits.

[0006] Frankincense extract has been used as an anti-inflammatory agent in traditional medicine, and 3-O-acetyl-11-carbonyl-β-boswellic acid (AKBA) is one of the reported anti-inflammatory components of the boswellic acid class of compounds. AKBA extracted from natural frankincense resin has been described in US patent US2003 / 0199581, etc. Studies have found that AKBA has a low inhibitory effect on T cell activity. US patent US2017 / 002039 reports that the compound CKBA is being investigated in China for the treatment of autoimmune diseases (such as psoriasis and vitiligo). CKBA is a candidate compound obtained by modifying AKBA as a lead compound, but its bioavailability is low, which affects the prospects for oral drug development.

[0007] In summary, further optimization of substances that are more effective in suppressing immune diseases is needed in this field. Summary of the Invention

[0008] The purpose of this invention is to provide the use of a class of pentacyclic triterpenoid derivatives and their pharmaceutically acceptable salts in the prevention and / or treatment of autoimmune diseases, wherein the application of these diseases, which are improved by the good immunomodulatory effects of Tc1, Tc17, and Th17 cells, is selected from autoimmune diseases related to multiple sclerosis, psoriasis, vitiligo, atopic dermatitis, inflammatory bowel disease, and alopecia areata. This invention also relates to methods for preparing such compounds.

[0009] In a first aspect of the invention, compounds of formula (I) or isomers thereof, solvates or precursors thereof, or pharmaceutically acceptable salts thereof are provided.

[0010]

[0011] R1 includes groups selected from the following group:

[0012] L is a linking group, which is a chemical bond, or a straight-chain or branched C1-C6 alkylene chain, wherein the straight-chain or branched C1-C6 alkylene chain may optionally be replaced by hydroxymethyl or methyl.

[0013] R2 is absent or includes a group selected from the group consisting of OH, NH2, or a substituted amino group, including straight-chain or cyclic amino groups, including groups selected from the group consisting of:

[0014]

[0015] In one or more embodiments, the compound comprises a subset selected from:

[0016]

[0017]

[0018] In another aspect of the invention, the use of the compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof, for the preparation of pharmaceutical compositions or kits for the prevention, relief or treatment of immune diseases is provided.

[0019] In one or more embodiments, the immune disease includes: autoimmune diseases, tumors.

[0020] In one or more embodiments, the autoimmune disease includes multiple sclerosis (MS) (which may include encephalitis, myelitis).

[0021] In one or more embodiments, the autoimmune disease includes: depigmenting skin diseases; preferably including: vitiligo, albinism (including chemical leukoderma, macular albinism), pityriasis alba, leprosy (with skin depigmentation), post-inflammatory hypopigmentation, progressive macular hypopigmentation, or tinea versicolor.

[0022] In one or more embodiments, the autoimmune disease includes: inflammatory skin diseases; preferably including: psoriasis, atopic dermatitis, alopecia areata.

[0023] In one or more embodiments, the autoimmune disease includes inflammatory bowel disease.

[0024] In one or more embodiments, the autoimmune disease includes: the immune disease includes: Tc1 cell overexpression (including dysregulation or disorder); Tc17 cell overexpression (including dysregulation or disorder); Th17 cell overexpression (including dysregulation or disorder).

[0025] In another aspect of the invention, the use of the compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof, for the preparation of reagents or kits for inhibiting the differentiation of immune cells (including: cells isolated in vitro or cell cultures), said immune cells comprising CD8 + T cells or CD4 + T cells.

[0026] In one or more embodiments, the inhibition of immune cell differentiation includes: inhibiting CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells.

[0027] In another aspect of the invention, a method for inhibiting the differentiation of immune cells (including: cells isolated in vitro or cell cultures) is provided, comprising: treating immune cells with a compound of formula (I) or an isomer thereof, a solvate or a precursor thereof, or a pharmaceutically acceptable salt thereof; said immune cells comprising CD8 + T cells or CD4 + T cells.

[0028] In one or more embodiments, the inhibition of immune cell differentiation includes: inhibiting CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells.

[0029] In one or more embodiments, the method for inhibiting immune cell differentiation is an in vitro method.

[0030] In one or more embodiments, the method for inhibiting immune cell differentiation is a non-therapeutic method.

[0031] In another aspect of the invention, a composition is provided comprising: the compound of formula (I) or an isomer thereof, a solvate or a precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically or biologically acceptable carrier;

[0032] In one or more embodiments, the composition is a pharmaceutical composition;

[0033] In one or more embodiments, the composition is a reagent composition, preferably an immune cell culture medium.

[0034] In one or more embodiments, the composition is a pharmaceutical composition, wherein the compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof, are present in an effective amount in the pharmaceutical composition; preferably, the effective amount is 0.01-50% by weight, for example, but not limited to, 0.01-5%, 0.03-3%, 0.05-1%, 20-30%, 40-50%, etc.; more preferably 0.03-30%; and even more preferably 0.05-10%.

[0035] In one or more embodiments, the composition is a pharmaceutical composition, the dosage form of which includes: powder, granule, tablet, pill, capsule, sustained-release preparation, controlled-release preparation, injection, infusion, and suspension.

[0036] In one or more embodiments, the pharmaceutical composition includes pharmaceutically or biologically acceptable excipients or diluents.

[0037] In another aspect of the invention, a kit or reagent kit is provided, comprising: the compound of formula (I) or an isomer thereof, a solvate or a precursor thereof, or a pharmaceutically acceptable salt thereof; or the composition thereof.

[0038] In another aspect of the invention, a method for preventing, alleviating, or treating an immune disease is provided, the method comprising: administering to a subject in need of treatment an effective amount of the compound of formula (I) or an isomer thereof, solvate or precursor thereof, or a pharmaceutically acceptable salt thereof.

[0039] Other aspects of the invention will be apparent to those skilled in the art from the disclosure herein. Attached Figure Description

[0040] Figure 1 CD8 + Results of experiments on T cell differentiation into Tc1 cells.

[0041] Figure 2 CD8 + Results of experiments on T cell differentiation into Tc17 cells.

[0042] Figure 3 CD4 + Results of experiments on T cell differentiation into Th17 cells.

[0043] Figure 4 Mouse EAE score curve. Detailed Implementation

[0044] Through in-depth research, the inventors have for the first time revealed a new class of pentacyclic triterpenoid derivatives, their structural characteristics, and preparation methods. The pentacyclic triterpenoid derivatives described in this invention possess excellent immunomodulatory effects and can be applied to the prevention and / or treatment of immune diseases.

[0045] the term

[0046] The term "isomer" as used in this article includes: geometric isomers, enantiomers, and diastereomers (such as cis-trans isomers and conformational isomers).

[0047] As used herein, the term "solvent" refers to a compound that carries solvent molecules; for example, the solvate may be a hydrate.

[0048] In this invention, the term "containing" indicates that various ingredients may be used together in the mixtures or compositions of this invention. Therefore, the terms "consistent with..." and "composed of..." are included in the term "containing".

[0049] In this invention, a "pharmaceuticalally acceptable" ingredient is a substance that is suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., has a reasonable benefit / risk ratio.

[0050] In this invention, a "pharmaceutically acceptable carrier" is a pharmaceutically or food-acceptable solvent, suspending agent, or excipient used to deliver the pentacyclic triterpenoid derivatives, isomers, solvates, precursors, or their pharmaceutically acceptable salts of this invention to animals or humans. The carrier may be liquid or solid.

[0051] Pentacyclic triterpenoid derivatives

[0052] This invention first provides a pentacyclic triterpenoid derivative as shown in structural formula (I):

[0053]

[0054] This invention also includes isomers, solvates, precursors, or pharmaceutically acceptable salts of the aforementioned pentacyclic triterpenoid derivatives, provided they also have the same or substantially the same function as the aforementioned pentacyclic triterpenoid derivatives. A "pharmaceutically acceptable salt" refers to a salt formed by the reaction of the compound with an inorganic acid, organic acid, alkali metal, or alkaline earth metal. These salts include (but are not limited to): (1) salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, carbonic acid, nitric acid, and phosphoric acid; and (2) salts formed with organic acids such as hydrobromic acid, citric acid, tartaric acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid. Other salts include salts formed with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium) in the form of esters, carbamates, or other conventional "prodrugs."

[0055] The compound has one or more asymmetric centers. Therefore, these compounds can exist as racemic mixtures, individual enantiomers, individual diastereomers, mixtures of diastereomers, or cis or trans isomers.

[0056] The term "precursor of the compound" refers to a compound that, when taken by an appropriate method, undergoes metabolism or chemical reaction in the patient's body to transform into a compound of structural formula (I), or a salt or solution of a compound of chemical structural formula (I).

[0057] As a preferred embodiment of the present invention, the compounds include: compound 1 to compound 20.

[0058] Those skilled in the art will understand that, after learning the structure of the compounds of the present invention, the compounds of the present invention can be obtained by a variety of methods well known in the art and using known raw materials, such as chemical synthesis or extraction from biological sources (e.g., animals or plants), all of which are included in the present invention.

[0059] As a preferred embodiment of the present invention, a method for preparing a class of pentacyclic triterpenoid derivatives is provided, comprising the following schemes:

[0060] Option 1:

[0061]

[0062] Starting with AKBA, intermediate A1 was obtained by acylation under thionyl chloride or oxalyl chloride conditions. Then, an amination reagent was added to obtain compound 1. Compound 1 was hydrolyzed under alkaline conditions to obtain compound 2.

[0063] Option 2:

[0064]

[0065] Starting with 3-hydroxy-11-carbonyl-β-boswellic acid (KBA), the carboxyl group of intermediate A2 is obtained by reacting with benzyl bromide under potassium carbonate or sodium carbonate conditions. A2 reacts with sodium hydride and iodomethane to generate intermediate A3. A3 is debenzylated under Pd / C hydrogenation to obtain the key intermediate A4. A4 is acylated under thionyl chloride or oxalyl chloride conditions to obtain intermediate A5. The addition of different amination reagents can yield compounds 3 to 16 or other derivatives with similar functional groups.

[0066] Option 3:

[0067]

[0068] The key intermediate A9 was obtained under similar reaction conditions as in Scheme 2. Compounds 17-18 or other similar functional group derivatives could be obtained by adding different amination reagents to the acylation reaction.

[0069] Option 4:

[0070]

[0071] The key intermediate A12 is obtained under similar reaction conditions as in Scheme 2. Compound 19 or other similar functional group derivatives can be obtained by adding an amination reagent to the acylation reaction.

[0072] Option 5:

[0073]

[0074] The key intermediate A15 is obtained under similar reaction conditions as in Scheme 4. Compound 20 or other similar functional group derivatives can be obtained by adding an amination reagent to the acylation reaction.

[0075] The synthesized compounds can be further purified by column chromatography, high performance liquid chromatography, and other methods.

[0076] application

[0077] The inventors discovered in their research that the pentacyclic triterpenoid derivatives of this invention can significantly inhibit CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells and significantly improve immune diseases.

[0078] The pentacyclic triterpenoid derivatives of this invention differ structurally from the compounds AKBA or CKBA disclosed in the prior art. The inventors' research shows that, using AKBA or CKBA as positive controls, the pentacyclic triterpenoid derivatives of this invention exhibit superior immunomodulatory effects on Tc1, Tc17, and Th17 cells. Some pentacyclic triterpenoid derivatives possess excellent bioavailability and readily cross the blood-brain barrier, facilitating oral administration and offering benefits for the relief or treatment of neurological disorders.

[0079] Based on the inventors' new discovery, the present invention provides the use of the aforementioned pentacyclic triterpenoid derivatives or their isomers, solvates, precursors, or pharmaceutically acceptable salts thereof for the preparation of medicaments or medicaments for the prevention, relief, or treatment of immune diseases.

[0080] Based on the inventors' new discovery, this invention provides the use of the aforementioned pentacyclic triterpenoid derivatives or their isomers, solvates, precursors, or pharmaceutically acceptable salts thereof for inhibiting CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells. The method described can be an in vitro method, that is, adjusting the proportion of different types in the T cell population in vitro, reducing Tc17 cells, Th17 cells, or Tc1 cells, thereby making the functional cells in the T cell population more dominant.

[0081] As one implementation method, the T cells can be treated under in vitro conditions to increase the proportion of functional cells, and then introduced into the body to exert an immunomodulatory effect.

[0082] Composition

[0083] The present invention also provides a pharmaceutical composition comprising: (a) an effective amount of the said pentacyclic triterpenoid derivative, or isomer thereof, solvate, precursor, or pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier or excipient.

[0084] In this invention, the pharmaceutical composition contains an effective amount of the pentacyclic triterpenoid derivative or its isomers, solvates, or precursors, or pharmaceutically acceptable salts thereof. For example, it may contain 0.001-50% by weight of the pentacyclic triterpenoid derivative or its pharmaceutically acceptable salt. Preferably, the pharmaceutical composition contains 0.01-20% by weight of the pentacyclic triterpenoid derivative or its pharmaceutically acceptable salt.

[0085] The dosage form of the pharmaceutical composition described in this invention can be diverse, as long as it enables the active ingredient to effectively reach the mammalian body. Examples include: powders, granules, tablets, pills, capsules, sustained-release formulations, controlled-release formulations, injections, infusions, and suspensions. Depending on the type of disease treated by the compounds according to this invention, those skilled in the art can choose a convenient dosage form.

[0086] From the perspective of ease of preparation and storage, preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules. From the perspective of ease of administration, preferred pharmaceutical compositions are oral formulations. The compounds of the present invention or pharmaceutical compositions thereof may also be stored in sterile instruments suitable for injection or infusion.

[0087] The effective dosage of pentacyclic triterpenoid derivatives as active ingredients can vary depending on the administration regimen and the severity of the disease being treated. However, generally, satisfactory effects are obtained when the compounds of the present invention are administered daily at a dose of about 0.01-100 mg / kg animal body weight, preferably in 1-3 separate doses daily, or in a sustained-release form. This dosage regimen can be adjusted to provide the best therapeutic response. For example, due to the urgency of the treatment condition, several separate doses may be administered daily, or the dose may be reduced proportionally.

[0088] The pharmaceutical composition may be placed in a kit. The kit may also include instructions for use that describe how to use the pharmaceutical composition.

[0089] The present invention also provides a composition (e.g., a culture medium) for cultivation, containing an effective amount of the said pentacyclic triterpenoid derivative, or an isomer thereof, solvate, precursor, or a pharmaceutically acceptable salt thereof. The composition for cultivation effectively inhibits CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells.

[0090] The composition for culture can be placed in a kit, or the composition can be added to an immune cell culture medium and then placed in a kit. The kit may also include instructions for use describing the cell culture method.

[0091] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0092] Example 1: Synthesis of N-(2-pyrrolidinylethyl)-3-O-acetyl-11-carbonyl-β-lactamamide compound 1

[0093]

[0094] Weigh 1.0 g (1.95 mmol) of 3-O-acetyl-11-carbonyl-β-boswellic acid and 5 mL of thionyl chloride into a 25 mL round-bottom flask. After heating and refluxing for 1 hour, remove the remaining thionyl chloride under reduced pressure to obtain intermediate compound A1, which is then dissolved in 5 mL of dichloromethane for later use.

[0095] Weigh 0.44 g (3.90 mmol) of 1-(2-aminoethyl)pyrrolidine, 0.59 g (5.85 mmol) of triethylamine, and 47.6 mg (0.39 mmol) of DMAP into 10 mL of dichloromethane solution. Cool the mixture to approximately 0 °C. While stirring, slowly add the dichloromethane solution containing the above A1 intermediate dropwise. After the addition is complete, allow the mixture to naturally warm to room temperature and react for 2 hours. Once the reaction is complete, add 10 mL of ice water to terminate the reaction. Extract the reaction solution with dichloromethane (10 mL x 3), wash once with saturated NaCl (10 mL), and finally dry the organic phase with anhydrous Na₂SO₄. After concentrating the organic layer, separate it by silica gel column chromatography (eluent: dichloromethane:methanol = 20:1–15:1) to obtain a white solid compound 1 (0.89 g, 75%). MS (ESI) m / z: [M+H] + =609.4. 1H-NMR(400M, CDCl3)δ:6.32(1H,brs),5.57(1H,s),5.38(1H,s),3.41-3.27(2H,m), 2.64-2.61(2H,t),2.54(5H,s),2.43(1H,s),2.36-2.29(1H,m),2.10(4H,m),1.95- 1.72(9H,m),1.65-1.39(7H,m),1.37(3H,s),1.33-1.25(3H,m),1.21(3H,s),1.17( 3H,s),1.15(3H,s),1.06-1.03(2H,m),0.96(3H,s),0.84(3H,s),0.83-0.82(3H,d). 13 C-NMR(400M, CDCl3)δ:199.06,175.43,170.17,164.64,130.55,73.80,60.34,59.01,53.63,50.34,46.53,45.05,43.75,40.89,39.30,39. 26,37.97,37.45,34.99,33.97,33.32,30.88,28.87,27.50,27.22,2 4.84,23.79,23.59,21.39,21.13,20.48,19.10,18.24,17.42,13.23.

[0096] Example 2: Synthesis of N-(2-pyrrolidinylethyl)-3-hydroxy-11-carbonyl-β-lactamamide compound 2

[0097]

[0098] Compound 1 (0.31 g, 0.51 mmol) and sodium hydroxide (0.057 g, 1.02 mmol) were weighed and placed in 10 mL of anhydrous ethanol. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction solution was poured into ice water (10 mL), extracted with dichloromethane (15 mL x 3), washed once with saturated NaCl (10 mL), and the organic layer was concentrated and separated by silica gel column chromatography (eluent: dichloromethane:methanol = 15:1 to 10:1) to obtain a white solid compound 1 (0.18 g, 62%). MS (ESI) m / z: [M+H] + =567.4. 1H-NMR(400M, CDCl3)δ:6.33(1H,t),5.56(1H,s),4.17(1H,s),3.41-3.23(2H,m), 2.62-2.59(2H,m),2.51-2.47(5H,m),2.39-2.36(2H,m),2.13-2.06(1H,m),1.93 -1.85(2H,m),1.82-1.72(7H,m),1.58-1.34(9H,m),1.33-1.24(9H,m),1.20(3H, s),1.14(3H,s),1.03-0.99(1H,m),0.95(3H,s),0.83(3H,s),0.81-0.79(3H,d). 13 C-NMR(400M, CDCl3)δ:199.32,176.64,164.68,130.59,70.82,60.47,59.01,53.64,53.60,48.79,47.29,45.11,43.80,40.92,39.30, 39.28,37.90,37.62,34.32,33.98,33.41,30.91,28.89,27.52,27.20,26.50,25.41,23.62,21.13,20.53,19.22,18.24,17.44,13.34.

[0099] Example 3: Synthesis of N-(2-dimethylaminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 3

[0100] Step 1: Synthesis of intermediate A2

[0101]

[0102] Weigh 4.70 g (10 mmol) of 3-hydroxy-11-carbonyl-β-boswellic acid (KBA), intermediate A1, 2.76 g (20 mmol) of K₂CO₃, and 25 mL of DMF into a 100 mL round-bottom flask. Slowly add benzyl bromide (2.56 g, 15 mmol) and react overnight at room temperature. After the reaction is complete as detected by TLC, pour the reaction mixture into 100 mL of ice water, extract with ethyl acetate (50 mL x 3), wash once with saturated NaCl (50 mL), and finally dry the organic phase with anhydrous Na₂SO₄. After concentrating the organic layer, separate it by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 10:1–6:1) to obtain a white solid intermediate A2 (4.58 g, 82%). MS (ESI) m / z: [M+H] + =561.4. 1H-NMR(400M, CDCl3)δ7.36-7.30(5H,m),5.53(1H,s),5.15-5.06(2H,q),4.12 (1H,s),2.48(1H,dt),2.40(1H,s),2.29(1H,m),2.11-2.04(1H,m),1.90-1.76 (3H,m),1.73-1.35(11H,m),1.32-1.24(8H,m),1.21-1.17(1H,m),1.11(3H,s) ,1.02-1.01(1H,m),0.97(3H,s),0.94(3H,s),0.81(3H,s),0.79-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ:199.60,176.49,165.06,135.73,130.49,128.54,128.42,128.22,70.62,66.23,60.34,59.02,48.92,47.54,45.0 8,43.79,40.93,39.28,37.39,33.96,33.94,32.87,30.92,28.86,27 .52,27.16,26.31,24.27,21.14,20.54,18.95,18.22,17.44,13.27.

[0103] Step 2: Synthesis of intermediate A3

[0104] Intermediate A2 (4.50 g, 8 mmol) and DMF (20 mL) were weighed and placed in a 100 mL round-bottom flask. The reaction solution was cooled to approximately 0°C with ice water. Under a nitrogen atmosphere, 60% NaH (1.28 g, 32 mmol) was slowly added and the mixture was stirred for 30 minutes. Iodomethane (2.27 g, 16 mmol) was slowly added dropwise to the above reaction solution. The mixture was slowly heated to room temperature and reacted for 2 hours. The reaction was detected as complete by TLC. After the reaction solution was cooled to approximately 0°C, a saturated NaCl solution (35 mL) was slowly added dropwise to quench the reaction. Then, ethyl acetate (50 mL * 3) was added for extraction, followed by washing once with saturated NaCl (50 mL). Finally, the organic phase was dried over anhydrous Na2SO4. After concentrating the organic layer, it was separated by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 15:1 to 10:1) to obtain a white solid intermediate A3 (4.33 g, 90%). MS (ESI) m / z: [M+H] + =575.4. 1H-NMR(400M, CDCl3)δ7.36-7.30(5H,m),5.52(1H,s),5.16-5.04(2H,q) ,3.50(1H,s),3.31(3H,s),2.41-2.39(2H,m),2.11-1.06(8H,m),1.53- 1.33(8H,m),1.30(3H,s),1.28(3H,s),1.24-1.13(2H,m),1.10(3H,s), 1.00(1H,m),0.97(3H,s),0.90(3H,s),0.81(3H,s),0.79-0.77(3H,d). 13 C-NMR(400M, CDCl3)δ199.62,176.67,164.78,135.83,130.57,128.52,128.41,128.17,80.11,66.19,60.30,59.03,57.02,49.63,48.00,45 .09,43.81,40.95,39.30,37.35,34.17,33.96,32.83,30.93,28.85,2 7.55,27.17,24.07,21.12,20.75,20.63,18.96,18.24,17.41,13.45.

[0105] Step 3: Synthesis of intermediate A4 (3-methoxy-11-carbonyl-β-boswellic acid)

[0106] Intermediate A3 (4.0 g, 6.97 mmol), Pd / C (10%, 500 mg), and ethanol (35 mL) were weighed. The mixture was purged three times with hydrogen and reacted at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was stopped, Pd / C was removed by filtration through a diatomaceous earth filter. The solvent was concentrated under reduced pressure, and the mixture was then separated by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 10:1–4:1) to obtain a white solid intermediate 4 (2.82 g, 85%). MS (ESI) m / z: [M+H] - =483.3. 1H-NMR(400M, CDCl3)δ5.54(1H,s),3.46(1H,s),3.32(3H,s),2.45-2.41( 2H,m),2.21-1.99(2H,m),1,92-1.82(2H,m),1.76-1.63(3H,m),1.54-1. 37(6H,m),1.32(3H,s),1.31(3H,s),1.27-1.21(4H,m),1.18(3H,s),1.1 3(3H,s),1.02-0.98(2H,m),0.94(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.76,183.28,165.06,130.50,79.86,60.36,59.02,57.05,49.52,47.76,45.13,43.83,40.95,3 9.29,37.47,34.09,33.98,32.84,30.94,28.88,27.54,27.19,24.21,21.15,20.68,20.62,18.82,18.40,17.43,13.39.

[0107] Step 4: Synthesis of intermediate A5

[0108] Weigh 100 mg (0.21 mmol) of 3-methoxy-11-carbonyl-β-boswellic acid and 5 mL of thionyl chloride into a 25 mL round-bottom flask. After heating and refluxing for 2 hours, remove the remaining thionyl chloride under reduced pressure to obtain intermediate compound A5, which is then dissolved in 3 mL of dichloromethane for later use.

[0109] Step 5: Synthesis of N-(2-dimethylaminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 3

[0110] N,N-dimethylethylamine (37 mg, 0.42 mmol), triethylamine (63.6 mg, 0.63 mmol), and DMAP (47.6 mg, 0.39 mmol) were weighed and placed in 6 mL of dichloromethane solution. The mixture was cooled to approximately 0 °C, and the dichloromethane solution of the above A5 intermediate was slowly added dropwise while stirring. After the addition was complete, the mixture was allowed to warm to room temperature and reacted for 2 hours. Upon completion of the reaction, 10 mL of ice water was added to terminate the reaction. The reaction solution was extracted with dichloromethane (10 mL x 3), washed once with saturated NaCl (10 mL), and finally dried over anhydrous Na₂SO₄. The concentrated organic layer was then separated by silica gel column chromatography (eluent: dichloromethane:methanol = 20:1–15:1) to obtain a white solid compound 3 (59.2 mg, 52%). MS (ESI) m / z: [M+H] + =555.4. 1 H-NMR(400M, CDCl3)δ5.53(1H,s),3.73(1H,s),3.52-3.41(2H,m),3.33(3H,s),3.1 0(3H,s),2.85-2.76(3H,m),2.49(2H,s),2.43-2.40(2H,t),2.12-2.04(2H,m),1.92 -1.76(3H,m),1.65-1.37(9H,m),1.34(3H,s),1.31(3H,s),1.26-1.25(3H,m),1.21( 3H,s),1.19(3H,s),1.01-0.97(2H,m),0.94(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.90,176.56,164.70,130.67,79.35,61.62,58.96,56.69,52.68,50.72,50.67,49.07,45.25,43.75,40.95,39.2 9,39.27,38.33,37.98,36.08,33.94,33.78,33.73,30.93,28.86,27 .55,27.21,22.87,21.43,21.14,20.67,20.35,18.46,17.41,16.05.

[0111] Example 4: Synthesis of N-(2-methylaminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 4

[0112]

[0113] Compound 4 (47 mg, 42%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =541.4. 1 H-NMR(400M, CDCl3)δ:6.21(1H,t),5.54(1H,s),3.52(1H,s),3.37-3.31(2H,m ),3.32(3H,s),2.77-2.73(2H,m),2.45(3H,s),2.44-2.40(2H,m),215-2.05(3H ,m),1.90-1.72(6H,m),1.54-1.36(8H,m),1.32(3H,s),1.24(3H,s),1.18(3H,s ),1.11(3H,s),1.02-0.98(2H,m),0.94(3H,s),0.82(3H,s),0.80-0.78(3H,s). 13 C-NMR(400M, CDCl3)δ199.43,177.13,164.61,130.63,80.31,60.40,59.01,57.02,50.14,49.50,47.90,45.11,43.83,40.94,39.31,3 9.30,38.47,37.57,35.76,34.51,33.98,33.18,30.93,28.86,27.54,27.18,25.10,21.13,20.97,20.62,19.40,18.29,17.42,13.51.

[0114] Example 5: Synthesis of N-(2-diethylaminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 5

[0115]

[0116] Compound 5 (65 mg, 48%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =583.5. 1H-NMR(400M, CDCl3)δ:6.38(1H,t),5.56(1H,s),3.55(1H,s),3.34(3H,s),3.30 -3.26(2H,q),2.59-2.49(6H,m),2.44-2.41(2H,m),2.17-2.07(2H,m),1.93-1. 69(6H,m),1.56-1.39(7H,m),1.35(3H,s),1.33-1.27(4H,m),1.25(3H,s),1.20 (3H,s),1.13(3H,s),1.03(6H,t),0.96(3H,s),0.83(3H,s),0.82-0.80(3H,d). 13 C-NMR(400M, CDCl3)δ199.40,176.72,164.50,130.65,80.42,60.41,59.01,57.01,50.98,49.47,47.75,46.31,45.11,43.84,40.95,3 9.30,37.58,36.67,34.55,33.99,33.28,30.93,28.88,27.55,27.21,25.21,21.13,20.92,20.62,19.53,18.38,17.43,13.51,11.98.

[0117] Example 6: Synthesis of N-(2-diethylaminopropyl)-3-methoxy-11-carbonyl-β-lactamamide compound 6

[0118]

[0119] Compound 6 (60 mg, 43%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =597.4. 1H-NMR(400M, CDCl3)δ:6.89(1H,t),5.52(1H,s),3.51-3.45(2H,m),3.37-3.34(1H,m) ,3.32(3H,s),3.13-2.94(6H,m),2.42-2.39(2H,m),2.15-1.99(4H,m),1.89-1.66(5H ,m),1.54-1.43(5H,m),1.40-1.36(6H,t),1.31-1.28(8H,m),1.25-1.22(2H,m),1.17 (3H,s),1.10(3H,s),1.02-0.99(2H,m),0.94(3H,s),0.81(3H,s),0.79-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.61,177.78,164.88,130.45,80.18,60.42,59.02,56.98,49.50,49.02,48.05,46.09,45.09,43.83,40.89,39. 25,39.22,37.42,36.41,34.37,33.94,33.03,30.87,28.84,27.46,2 7.10,25.00,23.67,21.09,20.55,19.57,18.38,17.38,13.70,8.44.

[0120] Example 7: Synthesis of N-(2-dimethylhydroxyethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 7

[0121]

[0122] Compound 7 (38 mg, 46%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =556.4. 1H-NMR(400M, CDCl3)δ:5.96(1H,t),5.54(1H,s),3.51(1H,s),3.44-3.87(1H,q),3.32 (3H,s),3.12-3.08(1H,q),2.60(1H,s),2.44-2.41(2H,m),2.13-2.05(2H,m),1.91-1. 69(6H,m),1.54-1.37(7H,m),1.32(4H,m),1.28(4H,m),1.24(3H,s),1.22(3H,s),1.1 8(3H,s),1.13(3H,s),1.02-0.98(2H,m),0.94(3H,s),0.81(3H,s),0.80-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.37,177.98,164.68,130.56,80.24,70.86,60.38,59.02,56.99,50.42,49.42,48.02,45.08,43.84,40.9 2,39.29,37.51,34.44,33.97,33.14,30.91,28.86,27.74,27.50,27.17,25.15,21.12,20.95,20.58,19.56,18.37,17.40,13.72.

[0123] Example 8: Synthesis of N-(1,3-dihydroxypropyl)-3-methoxy-11-carbonyl-β-lactamamide compound 8

[0124]

[0125] A similar synthetic method was used to prepare compound 1 to yield compound 8 (36 mg, 47%), a white solid. MS (ESI) m / z: [M+H] + =558.4. 1H-NMR(400M, CDCl3)δ:6.37-6.35(1H,d),5.52(1H,s),3.96-3.72(5H,m),3.50(1H ,s),3.41(1H,s),3.33-3.31(3H,m),2.42-2.38(2H,m),2.12-2.05(2H,m),1.94-1. 71(6H,m),1.54-1.37(7H,m),1.31(3H,s),1.27(3H,s),1.22-1.19(3H,s),1.16(3 H,s),1.13(3H,s),1.02-0.98(2H,m),0.94(3H,s),0.80(3H,s),0.79-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.74,177.75,165.22,130.43,80.13,63.68,63.23,60.39,59.04,57.04,52.08,49.44,48.04,45.13,43.86,40.92, 39.29,37.59,34.41,33.98,33.09,30.91,28.86,27.50,27.16,25.23 ,21.11,20.92,20.56,21.11,20.92,20.56,19.23,18.29,17.40,13.50

[0126] Example 9: Synthesis of N-(2-aminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 9

[0127]

[0128] Step 1: Synthesis of intermediate A6

[0129] A column-free crude intermediate A6 (150 mg) can be obtained by using a synthetic method similar to that used to prepare compound 1. A6 can be used directly in the next reaction.

[0130] Step 2: Synthesis of N-(2-aminoethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 9

[0131] Intermediate A6 (120 mg) was weighed and placed in a 25 mL round-bottom flask. Dichloromethane (5 mL) and trifluoroacetic acid (2 mL) were added, and the mixture was stirred at room temperature for 6 hours. After the reaction was complete, an ice-water mixture was added to dilute the solution. The pH was then adjusted to 7-8 with 1 M NaOH. Extraction was performed with ethyl acetate (15 mL x 3), followed by washing once with saturated NaCl (5 mL). Finally, the organic phase was dried over anhydrous Na₂SO₄, and the product (57 mg, 38%) was obtained by column chromatography. MS (ESI) m / z: [M+H] + =527.4. 1 H-NMR(400M, CDCl3)δ:6.21(1H,t),5.53(1H,s),3.52(1H,s),3.34-3.30(5H,m), 2.90-2.88(3H,t),2.50(2H,s),2.42-2.40(2H,m),2.15-2.05(2H,m),1.91-1.68 (5H,m),1.54-1.41(5H,m),1.33-1.29(5H,m),1.25(4H,m),1.22(1H,m),1.18(3H ,s),1.13(3H,s),1.02-0.98(2H,m),0.94(3H,s),0.81(3H,s),0.80-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.44,177.34,164.69,130.59,80.25,60.42,59.03,57.02,49.46,47.96,45.10,43.84,41.24,40.94,4 0.84,39.30,37.54,34.48,33.98,33.15,30.92,28.86,27.53,27.18,25.08,21.12,21.00,20.60,19.50,18.36,17.41,13.60.

[0132] Example 10: Synthesis of N-(2-piperazinylethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 10

[0133]

[0134] Compound 10 (28 mg, 36%) was prepared as a white solid using a synthetic method similar to that used for compound 9. MS (ESI) m / z: [M+H] + =596.5. 1H-NMR(400M, CDCl3)δ:6.16(1H,t),5.54(1H,s),3.54(1H,s),3.42-3.25(6H,m), 2.98(4H,t),2.55-2.50(6H,m),2.43-2.39(2H,m),2.13-2.05(2H,m),1.92-1.73 (6H,m),1.55-1.41(7H,m),1.32(3H,s),1.31-1.26(2H,m),1.23(3H,s),1.19(3H ,s),1.11(3H,s),1.03-1.01(2H,m),0.95(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.30,176.78,164.63,130.59,80.28,60.36,59.04,57.00,56.52,52.65,49.35,47.83,45.39,45.14,43.86,40.9 2,39.32,39.30,37.58,35.54,34.48,34.00,33.34,30.92,28.91,27 .52,27.23,25.25,21.12,20.88,20.59,19.54,18.52,17.42,13.59.

[0135] Example 11: Synthesis of N-(2-pyrrolithylethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 11

[0136]

[0137] Compound 11 (42 mg, 54%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =581.5;HRMS:Calcd for[C37H60N2O3+H] + ;581.4682;found,581.4686. 1H-NMR(400M, CDCl3)δ:6.29(1H,t),5.54(1H,s),3.55(1H,s),3.42-3.22(5H,m),2. 64-2.60(2H,m),2.53(4H,m),2.42-2.39(2H,m),2.13-2.05(2H,m),1.92-1.84(2H,m ),1.80-1.71(8H,m),1.54-1.37(6H,m),1.33-1.32(4H,m),1.28-1.24(5H,m),1.18( 3H,s),1.12(3H,s),1.02-0.98(2H,s),0.94(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.40,176.94,164.52,130.65,80.36,60.41,59.03,57.01,53.73,53.64,49.49,47.84,45.13,43.84,40.95,3 9.30,37.89,37.61,34.54,33.99,33.38,30.93,28.90,27.55,27.21,25.24,23.63,21.13,20.91,20.61,19.22,18.29,17.42,13.50.

[0138] Example 12: Synthesis of N-(2-pyrrolithopropyl)-3-methoxy-11-carbonyl-β-lactamamide compound 12

[0139]

[0140] Compound 12 (56 mg, 71%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =595.5. 1H-NMR(400M, CDCl3)δ:6.97(1H,t),5.52(1H,s),3.50-3.44(2H,m),3.38-3.3 3(4H,m),3.12-2.91(6H,m),2.42-2.39(2H,m),2.15-2.05(7H,m),2.00-1.65( 7H,m),1.54-1.36(7H,m),1.31(3H,s),1.27(3H,s),1.22(2H,m),1.17(3H,s) ,1.09(3H,s),1.02-0.99(2H,m),0.94(3H,s),0.82(3H,s),0.81-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.67,177.65,164.89,130.52,80.28,60.45,59.05,57.00,53.69,53.22,49.55,48.03,45.13,43.86,40.93,39.2 9,39.27,37.45,36.96,34.44,33.97,33.07,30.91,28.86,27.51,27 .15,25.44,24.99,23.25,21.11,20.58,19.55,18.39,17.40,13.71.

[0141] Example 13: Synthesis of N-(2-pyrrolidolyl)-3-methoxy-11-carbonyl-β-lactamamide compound 13

[0142]

[0143] Compound 13 (45 mg, 63%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =609.5. 1 H-NMR(400M, CDCl3)δ:6.10(1H,t),5.52(1H,s),3.51(1H,s),3.36-3.19(8H, m),3.08(2H,t),2.42-2.38(2H,m),2.14-2.05(6H,m),1.63-1.61(10H,m),1.5 4-1.37(7H,m),1.31-1.27(5H,m),1.35(3H,s),1.23-1.21(1H,m),1.17(3H,s) ,1.10(3H,s),1.02-0.99(2H,m),0.94(3H,s),0.81(3H,s),0.80-0.78(3H,d).13 C-NMR(400M, CDCl3)δ199.51,177.26,164.84,130.47,80.17,60.39,59.03,56.99,54.71,53.49,49.38,47.95,45.08,43.84,40.89,39.25,3 7.95,37.47,34.40,33.95,33.07,30.88,28.85,27.47,27.13,26.40, 25.03,23.30,22.86,21.09,21.04,20.56,19.60,18.39,17.38,13.69.

[0144] Example 14: Synthesis of N-(2-piperidinylethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 14

[0145]

[0146] Compound 14 (36 mg, 61%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =595.5. 1 H-NMR(400M, CDCl3)δ:5.53(1H,s),3.55(2H,s),3.33(3H,s),2.84(4H,b rs),2.42-2.39(2H,m),2.12-2.06(2H,m),1.90-1.68(11H,m),1.53-1.3 7(10H,m),1.31(3H,s),1.29(3H,s),1.25-1.19(3H,m),1.17(3H,s),1.0 9(3H,s),1.01-0.98(2H,m),0.94(3H,s),0.81(3H,s),0.80-0.78(3H,d). 13 C-NMR (400M, CDCl3) δ199.59,177.59,164.81,130.53,80.18,60.44,59.04,57.35,57.03,54.46,49.49,47.98,45.13,43.86,40.93,39.2 8,37.51,35.05,34.41,33.98,33.17,30.91,28.88,27.51,27.15,24 .84,23.86,22.81,21.12,20.99,20.58,19.43,18.38,17.41,13.60.

[0147] Example 15: Synthesis of N-(2-morpholinylethyl)-3-methoxy-11-carbonyl-β-lactamamide compound 15

[0148]

[0149] Compound 15 (45 mg, 58%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =597.5. 1 H-NMR(400M, CDCl3)δ:6.20(1H,t),5.54(1H,s),3.71-3.69(4H,s),3.55(1H,s),3.42 -3.36(1H,m),3.32(3H,s),3.30-3.24(1H,m),2.53-2.39(8H,m),2.14-2.05(2H,m),1 .92-1.70(6H,m),1.55-1.37(7H,m),1.32(3H,s),1.31-1.26(2H,m),1.24(3H,s),1.1 9(3H,s),1.11(3H,s),1.03-0.99(2H,m),0.95(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.26,176.77,164.56,130.61,80.29,67.16,60.36,59.03,57.00,56.45,53.22,49.36,47.82,45.14,43.85,40.9 2,39.32,39.29,37.59,35.42,34.48,33.99,33.37,30.92,28.89,27 .52,27.23,25.28,21.12,20.86,20.60,19.53,18.55,17.42,13.57.

[0150] Example 16: Synthesis of N-(2-morpholinylpropyl)-3-methoxy-11-carbonyl-β-lactamamide compound 16

[0151]

[0152] Compound 16 (43 mg, 47%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =611.5. 1H-NMR(400M, CDCl3)δ:6.28(1H,t),5.54(1H,s),3.78-3.74(4H,q),3.49(1H, s),3.39-3.25(5H,m),2.49-2.42(8H,m),2.17-2.05(2H,m),1.91-1.68(7H,m) ,1.54-1.33(8H,m),1.32(3H,s),1.29-1.25(2H,m),1.23(3H,s),1.18(3H,s) ,1.10(3H,s),1.03-0.98(2H,m),0.94(3H,s),0.82(3H,s),0.80-0.78(3H,d). 13 C-NMR(400M, CDCl3)δ199.40,176.75,164.63,130.60,80.33,66.63,60.40,59.02,57.83,56.99,53.91,49.49,47.84,45.07,43.83,40.93,3 9.30,39.29,38.99,37.48,34.50,33.98,33.12,30.92,28.87,27.52, 27.20,25.00,24.86,21.12,21.06,20.60,19.75,18.40,17.41,13.64.

[0153] Example 17: Synthesis of N-(2-pyrrolithylethyl)-3-(3-morpholinopropoxy)-11-carbonyl-β-lactamamide compound 17

[0154]

[0155] Step 1: Synthesis of intermediates A8 and A9

[0156] Crude intermediate A8 can be obtained using a synthetic method similar to that used to prepare intermediate A3, and white solid intermediate A9 (250 mg, 65%) can be obtained using a synthetic method similar to that used to prepare intermediate A4. MS (ESI) m / z: [M+H] + =598.4. 1H-NMR(400M, CDCl3)δ:10.34(1H,s),5.54(1H,s),3.77-3.76(5H,m),3.58-3.5 1(2H,m),3.35-3.30(1H,m),2.60-2.49(7H,m),2.44-2.39(2H,m),2.12-2.03( 2H,m),1.90-1.35(15H,m),1.32(3H,s),1.25(3H,s),1.23(1H,m),1.17(3H,s) ,1.15(3H,s),1.02-0.98(2H,m),0.95(3H,s),0.82(3H,s),0.82-0.79(3H,d).

[0157] 13 C-NMR (400M, CDCl3) δ199.87,180.71,164.88,130.53,78.38,66.67,66.05,6 5.96,60.44,60.30,59.00,56.03,53.03,52.78,49.54,47.68,45.15,43.75,4 0.94,39.28,39.25,37.43,34.47,33.95,32.94,30.91,28.85,27.53,27.18,26.04,24.73,21.68,21.14,20.55,18.97,18.55,18.41,17.44,13.68,11.76.

[0158] Step 2: Synthesis of N-(2-pyrrolithylethyl)-3-(3-morpholinopropoxy)-11-carbonyl-β-lactamamide compound 17

[0159] Compound 17 (28 mg, 52%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =694.5. 1H-NMR(400M, CDCl3)δ:6.32(1H,s),5.47(1H,s),3.64(4H,t),3.54(1H,s),3.36- 3.21(3H,m),2.65-2.62(4H,m),2.37-2.31(8H,m),2.05-1.98(2H,m),1.85-1.59( 14H,m),1.48-1.26(8H,m),1.25(3H,s),1.22-1.18(4H,m),1.16(3H,s),1.11(3H ,s),1.04(3H,s),0.96-0.91(2H,m),0.88(3H,s),0.75(3H,s),0.74-0.72(3H,d). 13 C-NMR(400M, CDCl3)δ199.45,177.20,164.58,130.62,78.30,67.01,60.47,59.00,56.26,54.02,53.80,49.42,47.92,45.12,43.77,40.9 2,39.29,37.55,34.66,33.98,33.41,30.91,28.89,27.52,27.26,27 .20,25.27,23.57,21.67,21.13,20.48,19.19,18.29,17.43,13.52.

[0160] Example 18: Synthesis of N-(2-piperidinylethyl)-3-(3-morpholinopropoxy)-11-carbonyl-β-lactamamide compound 18

[0161]

[0162] Compound 18 (40 mg, 48%) was prepared as a white solid using a synthetic method similar to that used for compound 17. MS (ESI) m / z: [M+H] + =708.5. 1H-NMR(400M, CDCl3)δ:6.40(1H,s),5.54(1H,s),3.72-3.70(4H,m),3.62-3.56( 2H,m),3.38-3.22(3H,m),2.44-2.38(12H,m),2.14-2.06(2H,m),1.93-1.65(8H ,m),1.58-1.37(13H,m),1.33(3H,s),1.29-1.24(4H,m),1.22(3H,s),1.19(3H, s),1.11(3H,s),1.03-0.99(2H,m),0.95(3H,s),0.82(3H,s),0.81-0.79(3H,d). 13 C-NMR (400M, CDCl3) δ199.44,176.78,164.50,130.64,78.38,67.02,60.5 3,59.02,56.59,56.28,54.13,53.80,49.38,47.85,45.13,43.80,40.93,3 9.30, 37.56, 35.79, 34.70, 33.99, 33.48, 30.92, 28.90, 27.54, 27.27, 27.23, 26.20, 25.38, 24.35, 21.67, 21.14, 20.49, 19.41, 18.44, 17.43, 13.55.

[0163] Example 19: Synthesis of N-(2-diethylaminoethyl)-3-(2-ethoxyethoxy)-11-carbonyl-β-lactamamide compound 19

[0164]

[0165] Step 1: Synthesis of intermediates A11 and A12

[0166] Crude intermediate A11 can be obtained using a synthetic method similar to that used to prepare intermediate A3, and white solid intermediate A12 (267 mg, 55%) can be obtained using a synthetic method similar to that used to prepare intermediate A4. MS (ESI) m / z: [M+H] + =543.4. 1H-NMR(400M, CDCl3)δ:5.55(1H,s),3.73-3.67(1H,m),3.62-3.46(6H,m),2. 46-2.42(2H,m),2.13-2.01(2H,m),1.92-1.83(2H,m),1.73-1.64(4H,q),1. 56,1.38(6H,m),1.35(3H,s),1.33(3H,s),1.27-1.21(6H,s),1.19(3H,s),1 .14(3H,s),1.03-1.00(2H,m),0.96(3H,s),0.83(3H,s),0.81-0.80(3H,d). 13 C-NMR (400M, CDCl3) δ199.73,183.24,164.90,130.53,78.38,69.84,68.50,66.49,60.37,59.02,49.27,47.75,45.13,43.80,40.9 5,39.29,37.44,34.18,33.97,32.86,30.94,28.87,27.55,27.19,24.15,21.67,21.14,20.56,18.80,18.40,17.42,15.27,13.40.

[0167] Step 2: Synthesis of N-(2-diethylaminoethyl)-3-(2-ethoxyethoxy)-11-carbonyl-β-lactamamide compound 19

[0168] Compound 19 (43 mg, 42%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =641.5. 1 H-NMR (400M, CDCl3) δ: 6.40 (1H, s), δ: 5.54 (1H, s), 3.72-3.68 (2H, m), 3.59- 3.46(5H,m), δ:3.27(2H,s), 2.58-2.50(6H,m), 2.42-2.39(2H,m), 2.17-2.0 6(2H,m),1.92-1.38(13H,m),1.32(5H,m),1.25(5H,m),1.21-1.19(6H,m),1 .12(3H,s),1.05-1.02(6H,t),0.95(3H,s),0.82(3H,s),0.81-0.79(3H,d). 13C-NMR (400M, CDCl3) δ199.45,176.87,164.47,130.63,78.80,69.84,68. 43,66.42,60.42,59.01,51.07,49.18,47.76,46.39,45.11,43.81,40.9 4,39.29,37.53,36.63,34.62,33.98,33.28,30.92,28.87,27.54,27.20 ,25.12,21.91,21.13,20.53,19.48,18.38,17.41,15.30,13.53,11.83.

[0169] Example 20: Synthesis of N-(2-diethylaminoethyl)-3-(2-methoxyethoxy)-11-carbonyl-β-lactamamide compound 20

[0170]

[0171] Step 1: Synthesis of intermediates A14 and A15

[0172] Crude intermediate A14 can be obtained using a synthetic method similar to that used to prepare intermediate A3, and white solid intermediate A15 (190 mg, 53%) can be obtained using a synthetic method similar to that used to prepare intermediate A4. MS (ESI) m / z: [M+H] + =529.4. 1 H-NMR(400M, CDCl3)δ:5.55(1H,s),3.73-3.68(1H,m),3.61(1H,t),3.56-3.54(2H,t), 3.51-3.46(1H,m),3.40(3H,t),2.47-2.42(2H,m),2.13-2.02(2H,m),1.93-1.83(2H,m) ,1.74-1.65(3H,m),1.56-1.38(7H,m),1.35(3H,s),1.33(3H,s),1.28-1.23(3H,m),1. 19(3H,s),1.14(3H,s),1.03-1.00(2H,m),0.96(3H,s),0.83(3H,s),0.82-0.80(3H,d). 13C-NMR (400M, CDCl3) δ199.68,182.94,164.87,130.54,78.50,72.13,68.47,60.35,59.02,58.95,49.28,47.73,45.12,43.80,40. 95,39.29,37.44,34.19,33.97,32.82,30.93,28.87,27.55,27.20,24.13,21.67,21.14,20.58,18.80,18.40,17.43,13.40,8.62.

[0173] Step 2: Synthesis of N-(2-diethylaminoethyl)-3-(2-methoxyethoxy)-11-carbonyl-β-lactamamide compound 20

[0174] Compound 20 (47 mg, 50%), a white solid, was prepared using a synthetic method similar to that used for compound 1. MS (ESI) m / z: [M+H] + =627.5. 1 H-NMR(400M, CDCl3)δ:6.35(1H,s),δ:5.54(1H,s),3.71-3.67(2H,m),3.54-3.45(3H,m ),δ:3.37(3H,s),3.28-2.24(2H,m),2.59-2.48(8H,m),2.17-2.06(2H,m),1.92-1.84( 1H,m),1.75-1.65(6H,m),1.55-1.43(6H,m),1.33-1.32(6H,m),1.24(3H,m),1.22-1.1 8(4H,m),1.11(3H,s),1.03-1.00(6H,t),0.94(3H,s),0.82(3H,s),0.81-0.79(3H,d). 13 C-NMR (400M, CDCl3) δ199.48,177.17,164.55,130.62,78.87,72.12,68.41,60.42,59.02,58.93,51.54,49.25,47.83,46.91,45.11,43.82,4 0.94,39.29,37.52,36.38,34.61,33.98,33.20,30.92,28.87,27.54, 27.19,24.98,21.93,21.13,20.54,19.47,18.39,17.43,13.54,11.15.

[0175] Example 21: In vitro effects of different compounds on mouse CD8+ Assay for T cell differentiation into Tc1 and Tc17 cells

[0176] Test cells and test animals: mice CD8 + T cells were isolated and extracted from the spleen of C57BL / 6 mice.

[0177] Preparation of solutions of different compounds:

[0178] Depending on the molecular weight of the different compounds, 1.5 mM solutions of different compounds were prepared using DMSO for later use.

[0179] Experimental steps:

[0180] (1) Select 6-8 week old C57BL / 6 mice, lyse red blood cells with red blood cell lysis buffer to obtain spleen single cell suspension;

[0181] (2) Using mice CD8 + T-cell magnetic bead enrichment kit was used to isolate and obtain CD8 + T cells. At 1.0 x 10⁻⁶ cells per well. 5 One cell was seeded in a 96-well plate;

[0182] (3) Under the induction conditions of anti-CD3, anti-CD28, IL-2 and IL-12, they differentiated into Tc1 cells, effector cells that produce IFN-γ;

[0183] (4) At the same time, 0.2 μL of different compound solutions with a concentration of 1.5 mM were added to each well, with a total volume of 200 μL / well (final concentration: 1.5 μM). The control group was given the corresponding volume of DMSO, and the blank control group was given only 200 μL of culture medium. Each group had three replicates.

[0184] (5) Under the induction conditions of anti-CD3, anti-CD28, TGF-β, IL-6, IL-23, IL-1β and anti-IFN-γ, they differentiated into Tc17 cells, which produce IL-17A.

[0185] (6) At the same time, 0.2 μL of different compound solutions with a concentration of 1.5 mM were added to each well, with a total volume of 200 μL / well (final concentration: 1.5 μM). The control group was given the corresponding volume of DMSO, and the blank control group was given only 200 μL of culture medium. Each group had three replicates.

[0186] (7) After culturing in a cell culture incubator (37℃, 5% CO2) for 72 hours, the proportion of Tc1 cells and Tc17 cells was detected by flow cytometry.

[0187] (8) Record the percentage of T cell subsets in CD8+ in different compound concentration groups. + The percentage of T cells was calculated by "inhibition percentage = (1 - flow cytometry value / mean flow cytometry value of DMSO group) × 100%". The average of the three inhibition percentages obtained for each group was calculated to obtain the average inhibition percentage of T cell subset differentiation for each compound concentration.

[0188] The compounds prepared in this invention are used as test compounds, with AKBA and CKBA as positive controls.

[0189]

[0190] For detailed results and statistical data, please refer to Tables 1 and 2. Figure 1 and Figure 2 .

[0191] Table 1, CD8 + Results of T cell differentiation into Tc1 cells

[0192]

[0193] Experimental conclusion: Under the experimental conditions, AKBA, CKBA, and various compounds have effects on mouse CD8. + The compounds in Table 1 inhibit T cell differentiation into Tc1 cells. + The inhibition rate of T cell differentiation into Tc1 cells was significantly better than that of CKBA.

[0194] Table 2: CD8 + Results of T cell differentiation into Tc17 cells

[0195]

[0196] Experimental conclusion: Under the experimental conditions, AKBA, CKBA, and various compounds have effects on mouse CD8. + All compounds in Table 2 inhibited T cell differentiation into Tc17 cells. + The inhibition rate of T cell differentiation into Tc17 cells was superior to that of CKBA.

[0197] Example 22: In vitro inhibition of CD4 by different compounds + T cell differentiation into Th17 cells assay

[0198] Test cells and test animals: mice CD4 + T cells were isolated and extracted from the spleen of C57BL / 6 mice.

[0199] Preparation of solutions for different compounds: Based on the molecular weight of different compounds, prepare 1.5 mM solutions of different compounds using DMSO for later use.

[0200] Experimental steps:

[0201] (1) Select 6-8 week old C57BL / 6 mice, lyse red blood cells with red blood cell lysis buffer to obtain spleen single cell suspension;

[0202] (2) Using mice CD4 + T-cell magnetic bead enrichment kit was used to isolate and obtain CD4 + T cells. At 1.0 x 10⁻⁶ cells per well. 5 One cell was seeded in a 96-well plate;

[0203] (3) Under the induction conditions of anti-CD3, anti-CD28, TGF-β, IL-6, IL-23 and anti-IFN-γ, they differentiated into Th17 cells, which produce IL-17A.

[0204] (4) At the same time, 0.2 μL of different compound solutions with a concentration of 1.5 mM were added to each well, with a total volume of 200 μL / well (final concentration: 1.5 μM). The control group was given the corresponding volume of DMSO, and the blank control group was given only 200 μL of culture medium. Each group had three replicates.

[0205] (5) After culturing in a cell culture incubator (37℃, 5% CO2) for 72 hours, the proportion of Th17 cells was detected by flow cytometry.

[0206] Record the percentage of Th17 cell subsets in CD8 in different compound concentration groups + The percentage of T cells was calculated by "inhibition percentage = (1 - flow cytometry value / mean flow cytometry value of DMSO group) × 100%". The average of the three inhibition percentages obtained for each group was calculated to obtain the average inhibition percentage of T cell subset differentiation for each compound concentration.

[0207] For detailed results and statistical data, please refer to Table 3. Figure 3 .

[0208] Table 3: CD4 + Results of experiments on T cell differentiation into Th17 cells

[0209]

[0210] Experimental conclusion: Under the experimental conditions, AKBA, CKBA, and various compounds have an effect on mouse CD4. + All compounds inhibited T cell differentiation into Th17 cells. Table 1 shows the compounds that inhibit CD4.+ The inhibition rate of T cell differentiation into Th17 cells is better than that of CKBA.

[0211] Example 23: Determination of Pharmacokinetic Properties in Rats

[0212] 1. Test Animals

[0213] Number of SD rats: 6, Body weight: 190 - 230 g.

[0214] Food and water supply: Free drinking water throughout the experiment. The gavage group was fasted for 12 hours before drug administration and fed 4 hours after drug administration. The intravenous injection group had free access to food and water.

[0215] 2. Dosage, Administration Method and Solvent of Different Compounds

[0216]

[0217] 3. Test Methods

[0218] In vivo pharmacokinetic experiments of compounds were carried out using SD rats. CKBA was administered by gavage at a dose of 5 mg / kg, and other test compounds were administered by gavage at a dose of 10 mg / kg. Blood samples were collected at the time points of 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h and 24 h, about 0.3 mL of blood sample was collected into heparinized Eppendorf tubes and centrifuged at 4000 rpm for 5 min at 4°C.

[0219] After plasma protein precipitation, WinNonlin 5.2 software was used for result analysis, and pharmacokinetic parameters were calculated according to the statistical moment theory.

[0220]

[0221]

[0222] Conclusion: The in vivo exposure of compounds 4 and 11 after oral administration is greater than 1000 hr*ng / mL, and their absolute bioavailability is higher than that of CKBA compound, showing good pharmacokinetic properties.

[0223] Example 24: Determination of Blood - Brain Barrier Permeability of Different Compounds in Rats

[0224] 1. Test Animals

[0225] The SPF - grade SD rats used in this experiment were purchased from Zhejiang Vital River Laboratory Animal Co., Ltd. Production License: SCXK(Zhe)2019 - 0001, Animal Certificate Number: 20240722Aazz0619000579, body weight 190 - 230 g.

[0226] 2. Dosage, administration method, and solvent for different compounds

[0227]

[0228] 3. Sample collection

[0229] Following drug administration, rats were euthanized by exsanguination at 5 min, 30 min, 2 h, 8 h, and 24 h. Whole blood was collected and placed in EDTA-K2 anticoagulant EP tubes on ice. The tubes were centrifuged at 1500–1600 g for 10 min at 4 °C to separate plasma. Plasma samples were transferred to new centrifuge tubes and stored at -90–-60 °C until analysis. Brain tissue was collected, and after removing surface blood vessels with filter paper, it was placed in self-sealing bags with corresponding numbers and stored at -90–-60 °C until analysis.

[0230] 4. Test methods

[0231] Take 50.0 μL of plasma / brain homogenate sample from male SD rats into a 96-well plate, add 300 μL of internal standard working solution (diazepam: 125 ng·mL⁻¹) or acetonitrile (for double blank samples and residual effect samples), vortex for 5 min to mix, centrifuge at 4000 rpm for 10 min at 4 °C, transfer 100 μL of supernatant to a 96-well plate containing 100 μL of water (containing 0.1% FA), vortex for 5 min and then inject for analysis.

[0232] 5. Pharmacokinetic Data Analysis

[0233] Pharmacokinetic parameters were calculated using a non-compartmental model in Phoenix WinNonlin 8.0 software. In this experiment, pharmacokinetic parameters were calculated based on the theoretical blood collection time and theoretical drug concentration specified in the protocol.

[0234]

[0235] Conclusion: The brain exposure of compound 11, 6126 hr*ng / m, is much greater than that of compound CKBA, 74.6 hr*ng / m, and the brain-blood ratio (B / P) is about 306 times that of compound CKBA, indicating that compound 11 has excellent brain permeability.

[0236] Example 25: Therapeutic effect of compound 11 on an experimental encephalomyelitis model induced by myelin.

[0237] 1. Experimental animals

[0238] Strain: C57 / bl6 mice; Age: 8-10 weeks; Source: Purchased from Shanghai Lingchang Biotechnology Co., Ltd., Certificate No.: 2013001822395.

[0239] 2. Test drug

[0240]

[0241] 3. Test Plan

[0242] 3.1 Establishment of the mouse EAE model

[0243] ① Dissolve 100g Bts in 20mL of incomplete Freund's adjuvant to prepare complete Freund's adjuvant;

[0244] ② Dissolve MOG in PBS to a final concentration of 3 mg / ml;

[0245] ③ Take 100 μL of complete Freund's adjuvant per mouse and mix it with MOG 1:1, then mix thoroughly by pipetting and blowing 500 times using an emulsion tube;

[0246] ④ Pertussis toxin (PT) was dissolved in PBS to a final concentration of 1 μg / ml, 200 μl per mouse;

[0247] ⑤ Ten 8-week-old C57 / bl6 mice were randomly divided into two groups of five each. The mice were anesthetized by intraperitoneal injection of 100 μl of 10% chloral hydrate solution, and 200 μl of PT was injected into the tail vein. Each mouse was also subcutaneously injected with 200 μl of emulsified MOG (two symmetrical points on the back).

[0248] ⑥ Two days later, administer another 200ul PT via the tail vein.

[0249] 3.2 Drug therapy

[0250] ① Dissolve compound 11 and dimethyl fumarate (DMF) in 0.5% CMC-Na at concentrations of 12 mg / ml and 6 mg / ml, respectively, and then sonicate to thoroughly mix the solution.

[0251] ② Treatment began on day 10 of MOG immunization. The experimental group received 100ul of 6mg / ml gavage twice daily, while the control group received 100ul of 0.5% CMC-Na and 6mg / ml of dimethyl fumarate (DMF) orally twice daily until day 30.

[0252] 4. Results and Statistical Analysis

[0253] Mouse EAE scores are shown below Figure 4 .

[0254] The results showed that compound 11 had a therapeutic effect comparable to that of the positive control drug dimethyl fumarate after administration.

[0255] According to an FDA announcement, patients using dimethyl fumarate are at risk of developing progressive multifocal leukoencephalopathy (PML), and this invention provides patients with more medication options.

[0256] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. The compound of formula (I) or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof, in, R1 includes groups selected from the following group: L is a linking group, which is a chemical bond, or a straight-chain or branched C1-C6 alkylene chain, wherein the straight-chain or branched C1-C6 alkylene chain may optionally be replaced by hydroxymethyl or methyl. R2 is absent or includes a group selected from the group consisting of OH, NH2, or a substituted amino group, including straight-chain or cyclic amino groups, including groups selected from the group consisting of:

2. The compound of formula (I) as claimed in claim 1, or an isomer, solvate, or precursor thereof, or a pharmaceutically acceptable salt thereof, characterized in that, The compounds include those selected from:

3. Use of the compound of formula (I) as claimed in claim 1 or 2, or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof, for the preparation of pharmaceutical compositions or kits for the prevention, relief or treatment of immune diseases.

4. Use of the compound of formula (I) as claimed in claim 3, or its isomers, solvates, or precursors, or pharmaceutically acceptable salts thereof, characterized in that, The immune diseases mentioned include: autoimmune diseases, tumors; Preferably, the autoimmune disease includes: Multiple sclerosis; Depigmenting skin diseases; preferably including: vitiligo, albinism, pityriasis alba, leprosy, post-inflammatory hypopigmentation, progressive macular hypopigmentation, or tinea versicolor; Inflammatory skin diseases; preferably including: psoriasis, atopic dermatitis, alopecia areata; Inflammatory bowel disease.

5. Use of the compound of formula (I) as claimed in claim 3, or its isomers, solvates, or precursors, or pharmaceutically acceptable salts thereof, characterized in that, The immune diseases mentioned include: Tc1 cell overexpression disease; Tc17 cell overexpression disease; Th17 cell overexpression disease.

6. Use of the compound of formula (I) according to claim 1 or 2, or isomers, solvates, or precursors thereof, or pharmaceutically acceptable salts thereof, for the preparation of a reagent or kit for inhibiting the differentiation of immune cells, said immune cells comprising CD8+. + T cells or CD4 + T cells; Preferably, the inhibition of immune cell differentiation includes: Inhibit CD8 + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells.

7. A method for inhibiting immune cell differentiation, comprising: Immune cells are treated with a compound of formula (I) as described in claim 1 or 2, or an isomer, solvate, or precursor thereof, or a pharmaceutically acceptable salt thereof; said immune cells include CD8. + T cells or CD4 + T cells.

8. The method for inhibiting immune cell differentiation as described in claim 7, characterized in that, The inhibition of immune cell differentiation includes: inhibiting CD8. + T cells differentiate into Tc1 cells, inhibiting CD8. + T cells differentiate into Tc17 cells, inhibiting CD4. + T cells differentiate into Th17 cells.

9. A composition comprising: a compound of formula (I) as claimed in claim 1 or 2, or an isomer thereof, a solvate or a precursor thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically or biologically acceptable carrier; Preferably, the composition is a pharmaceutical composition; Preferably, the composition is a reagent composition, and more preferably, it is an immune cell culture medium.

10. A medicine box or reagent kit, comprising: The compound of formula (I) as claimed in claim 1 or 2, or its isomers, solvates or precursors, or pharmaceutically acceptable salts thereof; Or the composition according to claim 9.

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