Carnosol derivative and application thereof in medicine for treating tumor cachexia

Abstract

The invention belongs to the field of pharmaceutical synthesis, and particularly relates to a carnosol derivative and application thereof in a medicine for treating tumor cachexia. The carnosol compound comprises a compound as shown in a formula (Y) or a pharmaceutically acceptable salt or optical isomer of the compound as shown in the formula (Y). The invention further provides a pharmaceutical composition for treating the cachexia disease and application of the pharmaceutical composition to preparation of a medicine for treating tumor cachexia, including muscle atrophy caused by tumor tissue, adipopenia caused by tumor tissue, appetite reduction caused by tumor tissue and inflammatory response caused by tumor tissue. And tumor cachexia caused by digestive tract related cancers, liver cancers, lung cancers and colon cancers. The invention also provides an application in preparation of drugs or inhibitors for treating or inhibiting amyotrophy, drugs or inhibitors for inhibiting or relieving fat cell lipolysis, and drugs or inhibitors for inhibiting or relieving weight loss or loss.

Description

[0001] This invention is a divisional application with application number 2023108498813, application date 2023-07-11, application type invention, and invention name “Carnosine derivatives and their use in drugs for the treatment of tumor cachexia”. Technical Field

[0002] The present invention belongs to the field of pharmaceutical synthesis, in particular to carnosol derivatives and applications thereof in drugs for treating tumor cachexia. Background Art

[0003] Tumor cachexia is a serious complication of cancer. It is a multifactorial syndrome characterized by a persistent loss of skeletal muscle (with or without fat), with clinical manifestations of anorexia, anemia, and involuntary weight loss. It involves metabolic changes in various tissues and organs. Skeletal muscle depletion is a key feature of tumor cachexia, with consequences including increased chemotherapy toxicity, complications from cancer surgery, and mortality. Tumor cachexia is highly associated with pancreatic, esophageal, gastric, lung, liver, and colon cancers, accounting for approximately half of all cancer deaths worldwide. It is most common in patients with advanced cancer. The prevalence of cachexia in cancer patients is as high as 50%-80%. The prevalence varies across tumor types, ranging from approximately 40% in patients with breast cancer and leukemia, 30% in lung cancer, approximately 50% in colon and prostate cancer, 41%-45% in liver cancer, approximately 70% in pancreatic cancer, and 80%-87% in gastric cancer. Weight loss is most common in patients with pancreatic or gastric cancer, exceeding 60%-80%, while the incidence of weight loss in patients with lung, colorectal, or prostate cancer is over 50%. Tumor cachexia not only severely impacts the effectiveness of chemotherapy, radiotherapy, and other medical treatments, but also impairs patients' quality of life and shortens their lifespan. The search for treatments, targets, or drugs to address this condition is urgent.

[0004] The currently known pathogenic mechanisms of tumor cachexia are primarily related to systemic inflammation, skeletal muscle loss and atrophy, fat loss, anorexia, multi-organ syndrome, and energy metabolism disorders. There are no specific treatments or medications. Currently, non-pharmacological palliative care, such as nutritional supplements and exercise therapy, is commonly used in clinical practice. However, the weight loss caused by tumor cachexia differs from the emaciation caused by malnutrition. In addition to the loss of adipose tissue, the more significant effect is the atrophy of skeletal muscle, the body's largest organ. This not only reduces myosin synthesis but also increases muscle degradation. Therefore, nutritional supplements can only partially alleviate the weight loss caused by cachexia but cannot address the underlying pathology. Exercise is difficult for patients who have already developed muscle atrophy caused by tumor cachexia. Therefore, current palliative care methods can only partially alleviate the symptoms of tumor cachexia but fail to achieve the goals of prevention and treatment.

[0005] Anamorelin is an oral, small-molecule (583.2 g / mol) ghrelin receptor agonist that is thought to improve tumor cachexia by increasing appetite. It has demonstrated excellent efficacy in a recent Phase III study and has received regulatory approval in Japan for the treatment of tumor cachexia. Medroxyprogesterone acetate is a synthetic progesterone and appetite stimulant approved for the treatment of unexplained anorexia, cachexia, or weight loss. Recently, Ruiz-Garcia et al. reviewed clinical trials of medroxyprogesterone acetate and found that, while it may cause weight gain, it did not improve quality of life.

[0006] In summary, while numerous drugs and antibodies have been developed in recent years to treat tumor cachexia, these efforts have struggled to advance due to poor clinical efficacy. Traditional Chinese medicine, with its thousands of years of history in treating chronic wasting diseases, offers important avenues for research into treating tumor cachexia.

[0007] Carnosol is a natural active diphenol diterpene compound and one of the active ingredients of plants such as rosemary and sage. Its molecular formula is C 20 H 26 O4, with a molecular weight of 330.42, has pharmacological activities such as anti-tumor, anti-inflammatory, and antioxidant effects. Studies have shown that carnosol can inhibit the proliferation of tumor cells in diseases such as breast cancer, colon cancer, and prostate cancer. In addition, carnosol has been identified as an effective antioxidant and is involved in the treatment of various diseases. Previously, we found that carnosol has a good anti-tumor cachexia effect and applied for a patent for "The use of carnosol compounds in the preparation of drugs for the treatment of cachexia diseases", patent number ZL 202010089693.1. However, the carnosol structure in this application is simple and cannot meet current needs. Summary of the Invention

[0008] The purpose of the present invention is to address the problems existing in the prior art, and on this basis, we have structurally modified carnosol, providing a synthesis of carnosol derivatives, such as carnosol derivatives with new structures (Carnosolanalogues), and studying their role in alleviating muscle atrophy and fat lipolysis caused by tumor cachexia. In the muscle direction, its mechanism is to inhibit the expression of p-p65 in a concentration-dependent manner, thereby inhibiting the overexpression of Atrogin-1, thereby alleviating muscle cell atrophy; on the other hand, carnosol derivatives increase the phosphorylation level of AKT in a concentration-dependent manner, promote the expression of MHC, MyoD and MyoG, and accelerate the synthesis of myotubular protein. In the fat direction, on the one hand, it can not only inhibit the activation expression of p-p65 and p-HSL in a concentration gradient, but also reduce the excessive lipolysis of 3T3-L1 adipocytes; on the other hand, carnosol derivatives inhibit the upregulation of p-AMPKα expression in 3T3-L1 adipocytes in a concentration-dependent manner, thereby avoiding excessive energy consumption and loss of fat. In summary, in vitro, carnosol derivatives can reduce protein degradation in muscle cells, promote protein synthesis in muscle cells and muscle cell growth and differentiation; reduce lipolysis and excessive energy consumption in adipocytes; in vivo, carnosol derivatives can alleviate the degradation of adipose tissue caused by tumor cachexia, reduce weight loss, and play a role in alleviating the symptoms of tumor cachexia. They are candidate compounds with great prospects for the treatment of tumor cachexia.

[0009] In order to achieve the above purpose of the invention, the technical solution of this application is:

[0010] Carnosol compounds, including compounds represented by formula (X) and / or formula (Y) or pharmaceutically acceptable salts or optical isomers of compounds represented by formula (X) and / or formula (Y),

[0011]

[0012] In the formula (X),

[0013] A is O or N; B is O or N; C is N;

[0014] When A is O, R1 is selected from H or a substituted or unsubstituted 5-7 membered aromatic heterocyclic group or a phenyl group or a substituted or unsubstituted C1-C 12 Alkyl, the substituent is selected from one or more of halogen, C1-C6 alkyl, cyano, trifluoromethyl, nitro, carboxyl, hydroxyl, hydroxymethyl, methoxy, methyl, amino, acetamido, methylsulfonyl, and methoxyamido, and the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, and S;

[0015] When A is N, R1 is selected from H or a substituted or unsubstituted 5-7 membered aromatic heterocyclic group or a phenyl group or a substituted or unsubstituted C1-C 12 The substituent is selected from one or more of halogen, C1-C6 alkyl, cyano, trifluoromethyl, nitro, carboxyl, hydroxyl, hydroxymethyl, methoxy, methyl, amino, acetylamino, methylsulfonyl, and methoxyamide; the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, and S; R2 is selected from H or substituted or unsubstituted C1-C 12 Alkyl, the substituent is selected from halogen, trifluoromethyl, 5-7 membered aromatic heterocyclic group, 3-6 membered cyclic group, 6 membered heterocyclic group, condensed ring group, substituted or unsubstituted phenyl, the substituent of phenyl is halogen, trifluoromethyl, the aromatic heterocyclic group and 6 membered heterocyclic group contain one or more heteroatoms selected from N, O, S.

[0016] In particular, some compounds exist as the following isomers:

[0017]

[0018] The general formula (Y) is shown below:

[0019]

[0020] In the general formula (Y),

[0021] A is N or O; D is selected from OH or OTf or methoxy or substituted or unsubstituted benzene ring; E is selected from OH or OTf or methoxy or substituted or unsubstituted benzene ring, and the substituent is halogen or methoxy;

[0022] R2 is selected from H or substituted or unsubstituted C1-C 12 Alkyl, the substituent is selected from halogen, trifluoromethyl, 5-7 membered aromatic heterocycle, 3-6 membered ring, 6 membered heterocyclic group, condensed ring, substituted or unsubstituted benzene ring, the substituent of the benzene ring is halogen, trifluoromethyl, the aromatic heterocyclic group and 6 membered heterocyclic group contain one or more heteroatoms selected from N, O, S;

[0023] R3 is selected from OH or a substituted aliphatic group, wherein the substituent is selected from phenyl and a 6-membered aromatic heterocyclic group, wherein the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, and S.

[0024] Preferably, A=O, B=O, C=N, R1= Preferably, in the general formula (X), A=O, B=O, C=N, and its structural formula is as follows:

[0025]

[0026] The synthesis method is as follows:

[0027]

[0028] Preferably, in the general formula (X), A=N, B=O, C=N, and its structural formula is as follows:

[0029]

[0030] The synthesis method is as follows:

[0031]

[0032] Preferably, in the general formula (Y), A=N, D=OH, E=OH, and its structural formula is as follows:

[0033]

[0034] The synthesis method is as follows:

[0035]

[0036] Preferably, in the general formula (Y), A=O, D=OMe, E=OMe, and its structural formula is as follows:

[0037]

[0038] The synthesis method is as follows:

[0039]

[0040] The present invention also proposes a new structural formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one (Compound 1) with the following structural formula:

[0041]

[0042] Another invention objective of the present application is to protect the use of carnosine compounds represented by general formula (X) and general formula (Y) in the preparation of drugs for treating cachexia.

[0043] Furthermore, the drug is a drug that can be used for humans and / or animals.

[0044] Furthermore, the carnosol compounds represented by general formula (X) and general formula (Y) can be used in food, nutritional supplements and / or health products for humans and / or animals.

[0045] Furthermore, the dosage form of the product for treating cachexia is capsule, tablet, oral preparation, microcapsule preparation, injection, ointment, spray or suppository.

[0046] Furthermore, the product for treating cachexia is administered by injection, oral administration, parenteral administration, inhalation spray or transdermal administration.

[0047] Furthermore, the product for treating cachexia is a product for treating tumor cachexia.

[0048] In the application of the present invention, the cachexia disease includes tumor cachexia; the tumor is a solid tumor. The tumor cachexia includes, but is not limited to, muscle atrophy caused by tumor tissue, fat loss caused by tumor tissue, decreased appetite caused by tumor tissue, inflammatory response caused by tumor tissue, and tumor cachexia caused by digestive tract cancers, liver cancer, lung cancer, and colon cancer.

[0049] Furthermore, the product comprises the compound of formula (X) and / or formula (Y) in an amount of 0.001-100 wt.-% based on the total dry weight of the composition.

[0050] Furthermore, the general formula (X) and / or general formula (Y) is contained in an amount of 0.01 μg-100 mg / kg body weight / day.

[0051] The present invention also provides a method for treating cachexia, wherein the dosage of (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one (Compound 1) represented by formula (1) is 0.01 μg-100 mg / kg body weight / day.

[0052] Furthermore, the product also includes a protein source, a fat source and / or a carbohydrate source.

[0053] Furthermore, the product is selected from the group consisting of nutritionally balanced foods, complete nutritional formulas, dairy products, frozen or room temperature stable beverages, soups, nutritional bars, desserts, pet foods, pharmaceutical compositions and combinations thereof.

[0054] The present invention also proposes the use of a novel structural formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one (Compound 1) in the preparation of a drug for treating cachexia, which has a good anti-cachexia effect.

[0055] The present invention also provides a preparation method of the formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one (Compound 1). The reaction process and reaction formula of the preparation method are as follows:

[0056] The present invention's formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one (Compound 1) can be used to prepare an anti-cachexia drug. The cachexia includes, but is not limited to, progressive wasting syndrome caused by tumors, chronic kidney disease, chronic obstructive pulmonary disease, chronic heart failure, and AIDS. The drug is a drug for humans and / or animals.

[0057] The term "carnosol compound" used in the present invention refers to a single ingredient or a composition composed of other pharmaceutically acceptable ingredients for combined action.

[0058] The term "other pharmaceutically acceptable ingredients" used in the present invention refers to drugs that have no antagonistic effect on general formula (X) and general formula (Y), and can also be any one or more pharmaceutically acceptable excipients.

[0059] The term "cachexia" as used in the present invention refers to cachexia induced by solid tumors.

[0060] The term "solid tumor" as used in the present invention refers to primary or secondary solid tumors such as digestive tract cancer, liver cancer and lung cancer.

[0061] The muscle atrophy described in the present invention is muscle atrophy induced by tumor cachexia.

[0062] The fat degradation described in the present invention is fat degradation caused by tumor cachexia.

[0063] The administration of the carnosol compounds of general formula (X) and general formula (Y) of the present invention or the compositions thereof with other pharmaceutically acceptable ingredients is by injection, oral administration, parenteral administration, inhalation spray or transdermal administration.

[0064] In the present invention, the carnosol compounds of general formula (X) and general formula (Y) are used in the preparation of a medicament for treating cachexia. The medicament can be used in humans and / or animals. The dosage of the carnosol compounds of general formula (X) and general formula (Y) is 0.01 μg-100 mg / kg body weight / day. The medicament includes a drug or inhibitor for treating or inhibiting muscle atrophy, a drug or inhibitor for inhibiting or alleviating adipocyte lipolysis, or a drug or inhibitor for inhibiting or alleviating weight loss or reduction.

[0065] The third invention object of this application is to protect a pharmaceutical composition for treating tumor cachexia, wherein the composition contains a carnosine compound represented by general formula (X) and / or general formula (Y), and / or other pharmaceutically acceptable ingredients.

[0066] Furthermore, the other pharmaceutically acceptable ingredients may be drugs that have no antagonistic effect on the caryophyllene compounds, or may be any one or more pharmaceutically acceptable excipients.

[0067] The product dosage forms of the composition include, but are not limited to, capsules, tablets, oral preparations, microcapsule preparations, injections, ointments, sprays, or suppositories. The administration methods of the composition include, but are not limited to, injection, oral administration, parenteral administration, inhalation spray, or transdermal administration. The carnosin compounds represented by general formula (X) and / or general formula (Y) account for 0.001-100 wt.% of the total dry weight of the composition.

[0068] In the application of the present invention, the cachexia disease includes tumor cachexia; the tumor is a solid tumor. Tumor cachexia includes, but is not limited to, muscle atrophy caused by tumor tissue, fat loss caused by tumor tissue, decreased appetite caused by tumor tissue, inflammatory response caused by tumor tissue, and tumor cachexia caused by digestive tract cancers, liver cancer, lung cancer, and colon cancer.

[0069] The present invention also provides a method for treating cachexia, wherein the dosage of the carnosin compounds represented by general formula (X) or general formula (Y) is 0.01 μg to 100 mg / kg body weight / day. The administration methods include, but are not limited to, injection, oral administration, parenteral administration, inhalation spray, or transdermal administration.

[0070] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the carnosin compounds represented by the general formula (X), the general formula (Y) and pharmaceutically acceptable salts thereof.

[0071] The present invention also provides a pharmaceutical composition for treating cachexia, wherein the composition contains the formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one, and / or other pharmaceutically acceptable ingredients. The product dosage form of the pharmaceutical composition includes but is not limited to capsules, tablets, oral preparations, microcapsule preparations, injections, ointments, sprays or suppositories, etc. The product administration method of the pharmaceutical composition includes but is not limited to injection, oral, parenteral, inhalation spray or transdermal administration, etc. The (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(epoxymethane)phenanthren[3,4-d]oxazol-12-one represented by the formula (1) accounts for 0.001-100 wt.% of the total dry weight of the composition.

[0072] The present invention further proposes that the composition product further comprises a protein source, a fat source, and / or a carbohydrate source, and can be selected from nutritionally balanced foods, complete nutritional formulas, dairy products, frozen or room temperature stable beverages, soups, nutritional bars, desserts, pet foods, pharmaceutical compositions, and combinations thereof. The product contains carnosyl phenol compounds represented by general formula (X) and general formula (Y).

[0073] The present invention also proposes that the carnauba phenol compounds represented by the general formula (X) and the general formula (Y) include but are not limited to medicines for humans and / or pets, foods for humans and / or pets, nutritional supplements and / or health products.

[0074] The present invention also provides a product for treating cachexia using the carnosin compounds represented by general formula (X) or (Y) as a single ingredient or in combination with other pharmaceutically acceptable ingredients. The other pharmaceutically acceptable ingredients may be drugs that have no antagonistic effect on the compounds, or any one or more pharmaceutically acceptable excipients.

[0075] Compared with the prior art, the present invention has the following beneficial effects:

[0076] We have structurally modified carnosol and discovered that the newly modified carnosol derivatives (carnosol analogues) in this patent have the effect of alleviating muscle atrophy and fat lipolysis caused by tumor cachexia. In terms of muscle, its mechanism is, on the one hand, to inhibit the expression of p-p65 in a concentration-dependent manner, thereby inhibiting the overexpression of Atrogin-1, thereby alleviating muscle cell atrophy; on the other hand, carnosol derivatives upregulate AKT phosphorylation levels in a concentration-dependent manner, promote the expression of MHC, MyoD, and MyoG, and accelerate myotubular protein synthesis. In terms of fat, on the one hand, it can not only inhibit the activation expression of p-p65 and p-HSL in a concentration-dependent manner, reducing excessive lipolysis in 3T3-L1 adipocytes; on the other hand, carnosol derivatives inhibit the upregulation of p-AMPKα expression in 3T3-L1 adipocytes in a concentration-dependent manner, thereby preventing excessive energy consumption and loss of fat. In summary, in vitro, carnosol derivatives can reduce protein degradation in muscle cells, promote protein synthesis in muscle cells and muscle cell growth and differentiation; reduce lipolysis and excessive energy consumption in adipocytes; in vivo, carnosol derivatives can alleviate the degradation of adipose tissue caused by tumor cachexia, reduce weight loss, and play a role in alleviating the symptoms of tumor cachexia. They are candidate compounds with great prospects for the treatment of tumor cachexia. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Figure 1 This is the HE staining picture of the control group in Table 2 in Example 6.

[0078] Figure 2 This is the HE staining image of the model group in Table 2 in Example 6.

[0079] Figure 3-54 They are HE staining images of experimental groups 1-52 in Table 2 in Example 6.

[0080] Figure 55 -58 are the experimental results of the effects of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 in Example 7 on the activity of C2C12 muscle cells.

[0081] Figure 59 This is the HE staining image of the control group 1 in Table 4 in Example 8.

[0082] Figure 60 This is the HE staining image of model group 1 in Table 4 in Example 8.

[0083] Figure 61 -76 are HE staining images of experimental groups 1-14 in Table 4 in Example 8, respectively.

[0084] Figure 77 This is a statistical graph showing the effect of I-4-6 and II-6-2 in alleviating C2C12 muscle cell atrophy in Example 8.

[0085] Figure 78 This is a statistical graph showing the effect of II-8-1 and II-8-2 in Example 8 on the alleviation of C2C12 muscle cell atrophy.

[0086] Figures 79-82 These are the protein immunoblotting experimental results of I-4-6, II-6-2, II-8-1, and II-8-2 in Example 8 in the C2C12 cell muscular atrophy model.

[0087] Figure 83 -86 are the experimental results of the effects of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 on the activity of 3T3-L1 adipocytes in Example 9.

[0088] Figure 87 This is the Oil Red O staining image of the control group 1 in Table 7 in Example 10.

[0089] Figure 88 This is the Oil Red O staining image of C26 model group 1 in Table 7 in Example 10.

[0090] Figures 89-91 This is the Oil Red O staining image of experimental groups 1-3 in Table 7 in Example 10.

[0091] Figure 92 This is the Oil Red O staining image of the control group 2 in Table 7 in Example 10.

[0092] Figure 93 This is the Oil Red O staining image of C26 model group 2 in Table 7 in Example 10.

[0093] Figure 94 -96 are Oil Red O staining images of experimental groups 4-6 in Table 7 in Example 10.

[0094] Figures 97-98 They are semi-quantitative results of the Oil Red O staining experiment in which I-4-6 and II-6-2 alleviate lipolysis in 3T3-L1 adipocytes in Example 10.

[0095] Figure 99 This is the result of protein immunoblotting experiment of I-4-6 in Example 10 in 3T3-L1 cell lipolysis model.

[0096] Figure 100 These are the PK test results of I-5-3, I-4-22, I-4-6, II-6-2 and II-8-1 in Example 11.

[0097] Figure 101 This is a graph showing the body weight of tumor-bearing mice in the experiments I-4-6 and II-6-2 in Example 12.

[0098] Figure 102This is a graph showing the weight of mice after tumor removal in the experiments I-4-6 and II-6-2 in Example 12.

[0099] Figure 103 This is a graph showing the changes in tumor volume in mice tested in experiments I-4-6 and II-6-2 in Example 12.

[0100] Figure 104 This is a graph showing the tumor weights of mice in the experiment I-4-6 in Example 12.

[0101] Figure 105 This is the anatomical diagram of the tumor in the mouse experiment I-4-6 in Example 12.

[0102] Figure 106 This is a graph of the gastrocnemius muscle mass of mice in the experiment I-4-6 in Example 12.

[0103] Figure 107 This is the anatomical diagram of the gastrocnemius muscle of the mouse in the experiment I-4-6 in Example 12.

[0104] Figure 108 This is a graph of epididymal fat mass in mice from Experiment 1-4-6 in Example 12.

[0105] Figure 109 This is the anatomical diagram of the epididymal fat of the mouse in Experiment Ⅰ-4-6 in Example 12. DETAILED DESCRIPTION

[0106] The present invention is further illustrated below with reference to the following examples, which are in no way intended to limit the present invention. In all examples, the final product was purified using an LC-3000 high-performance liquid chromatograph developed by our research group at Sichuan University of Science and Chemical Technology. The liquid chromatography column used was an Extend-C18, 0.5 μm, 30 x 100 mm. 1 H-NMR was measured using a Bruker Avance III 600 MHz nuclear magnetic resonance instrument from Sichuan University of Science and Chemical Technology, and chemical shifts are expressed in δ (ppm). Mass spectra were measured using an AB Sciex Triple TOF 5600 mass spectrometer from Kunming Institute of Botany.

[0107] The synthesis of compounds Ⅰ-2-1 and Ⅰ-3-1 can be referred to the literature (Journal of Medicinal Chemistry, 2019, 62(23), 10867-10896); among them, carnosic acid (Ⅰ-1) was purchased from Shaanxi Guanchen Biotechnology Co., Ltd., 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, aldehyde compounds, tetrakistriphenylphosphine palladium, boric acid derivatives, and sodium cyanoborohydride were purchased from Anhui Zesheng Technology Co., Ltd., and sodium borohydride was purchased from Chengdu Cologne Chemical Co., Ltd., and iodomethane, 2,2-dimethoxypropane, pyridinium p-toluenesulfonate, chromium trioxide, ammonium hydrochloride, tetraisopropyl titanate, amine compounds, and trifluoroacetic acid were purchased from Shanghai Titan Technology Co., Ltd.

[0108] In the present invention, the pharmacodynamic test method used is a method well known and recognized by technical researchers and developers in this field.

[0109] In the present invention, the C2C12 cells (mouse myoblasts), 3T3-L1 cells (mouse adipocytes), and C26 cells (mouse colon cancer cells) used were purchased from the Cell Bank of the Committee for Type Culture Collection of the Chinese Academy of Sciences. BALB / c mice were purchased from Shanghai Jihui or Slake Co., Ltd.

[0110] Carnosol was purchased from Shanghai Standard Technology Co., Ltd.

[0111] Carnosol derivatives were provided by Sichuan University of Science & Engineering.

[0112] FBS (fetal bovine serum) was purchased from BI.

[0113] Horse serum was purchased from Gibco, and high-glucose DMEM medium was purchased from Hyclone.

[0114] RPMI-1640 medium was purchased from Hyclone. Phenol red-free high-glucose DMEM medium was also purchased from Hyclone.

[0115] P / S double antibody (penicillin-streptomycin mixture) was purchased from Hyclone. Dexamethasone was purchased from Sigma-Aldrich.

[0116] IBMX (3-isobutyl-1-methylxanthine), a broad-spectrum phosphodiesterase inhibitor, was purchased from Sigma–Aldrich.

[0117] Human recombinant insulin was purchased from Shanghai Jinmai Biotechnology.

[0118] The specific embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0119] Synthesis of target compounds

[0120] Example 1: Synthesis of oxazole carnosol

[0121]

[0122] Step 1: Preparation of I-2

[0123] I-1 (10.0 g, 30.5 mmol) and 100 mL of dry 1,4-dioxene were added to a 250 mL round-bottom flask and stirred at room temperature until completely dissolved. Dichlorodicyanobenzoquinone (DDQ) (7.61 g, 33.5 mmol) was slowly added and stirred at room temperature for 0.5 h. The reaction was then determined to be complete by TLC. The reaction mixture was filtered under reduced pressure, and the filter cake was washed with 50 mL of dichloromethane. 20 g of column chromatography silica gel was added to the filtrate and the mixture was evaporated under reduced pressure until solvent-free. The residue was ground into a powder and poured into a Buchner funnel. The powder was washed with 200 mL of dichloromethane:ethyl acetate (50:1). The filtrate was evaporated to dryness under reduced pressure. 30 mL of anhydrous ethanol was added to the residue and heated under reflux to completely dissolve the solid. The mixture was then slowly cooled to crystallize. After filtration, I-2 (7.7 g, 77.5% yield) was obtained. 1 H-NMR (600MHz, Acetone-d6) δ6.77 (s, 1H), 5.43 (s, 1H), 3.30 (dq, J = 13.6, 6.8Hz, 1H), 2.85-2. 79(m,1H),2.54(td,J=13.8,4.4Hz,1H),2.25-2.16(m,1H),1.96(dt,J=13.7,3.2Hz,1H),1.85( ddd,J=13.8,10.7,1.5Hz,1H),1.69(dd,J=10.7,5.8Hz,1H),1.57(dq,J=10.5,3.7Hz,1H),1.54 -1.47(m,1H),1.32(td,J=13.4,3.2Hz,1H),1.19(dd,J=6.8,3.9Hz,6H),0.88(d,J=3.5Hz,6H).

[0124] Step 2: Preparation of I-3-1

[0125] I-2 (3.0 g, 9.1 mmol) was added to a 100 mL round-bottom flask. Under nitrogen, 50 mL of dry 1,4-dioxene was added and stirred at room temperature until completely dissolved. Dichlorodicyanobenzoquinone (DDQ) (2.27 g, 10.01 mmol) was slowly added and stirred at room temperature for 0.5 h. The reaction was complete by TLC. The reaction mixture was filtered under reduced pressure, and the filter cake was washed with 20 mL of 1,4-dioxene. The solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether:dichloromethane = 2:3) to obtain I-3-1 (2.61 g, 87.0% yield) as a green solid. ESI-MS m / z: 329 [M+H] + .

[0126] 1 H NMR(600MHz,Chloroform-d)δ6.70(s,1H),5.22(s,1H),2.96(hept,J=7.8,7.3Hz,1 H),2.67(d,J=14.3Hz,1H),2.33–2.19(m,2H),1.99(ddd,J=14.1,10.7,1.4Hz,1H), 1.88(qt,J=13.9,3.3Hz,1H),1.65(ddd,J=16.7,10.3,4.7Hz,2H),1.52(d,J=14.8H z,1H),1.22(td,J=13.6,3.5Hz,1H),1.12(d,J=6.9Hz,6H),0.88(d,J=21.1Hz,6H). 13 C NMR(150MHz,Chloroform-d)δ179.19,176.26,173.43,152.71,151.06,135.54,129.76,76 .05,48.70,44.84,40.48,34.46,32.00,27.83,27.78,27.00,21.46,21.33,19.36,18.26.

[0127] Step 3: Preparation of I-4

[0128] Preparation of compounds I-4-1 to I-4-30: Under nitrogen, add I-3-1 (200 mg, 0.61 mmol), ammonium acetate (470 mg, 6.1 mmol), the corresponding aldehyde (1.22 mmol), and glacial acetic acid (10 mL) to a 25 mL single-necked flask. Heat to 120°C and reflux for 5-20 h. TLC confirms the reaction is complete. Cool to room temperature, remove acetic acid under reduced pressure, add saturated sodium bicarbonate solution (20 mL), and extract with ethyl acetate (20 mL x 3). Dry the extract with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure. The residue is purified by silica gel column chromatography to yield the corresponding products I-4-1 to I-4-30.

[0129]

[0130] Compound Ⅰ-4-1: white solid, HR-MS m / z: C 21 H 25 NO3,362.1727[M+Na] + (calculated value), 362.1729[M+Na] + (measured value), 1 H NMR (600MHz, CDCl3) δ8.08(s,1H),7.16(s,1H),5.66(dd,J=4.2,1.8Hz,1H),3.61(p,J=7.2Hz,1H),3.00(d ,J=13.8Hz,1H),2.43(td,J=13.8,4.8Hz,1H),2.32(ddd,J=13.8,6.0,4.2Hz,1H),2.10(qt,J=13.8,3.6Hz ,1H),1.96(ddd,J=13.8,10.8,1.8Hz,1H),1.76(dt,J=13.8,3.6Hz,1H),1.69(dd,J=10.2,6.0Hz,1H),1.6 1(d,J=15.0Hz,1H),1.37(dd,J=6.6,3.0Hz,6H),1.34–1.28(m,1H),1.26(s,2H),0.97(s,3H),0.88(s,3H). 13 C NMR (150MHz, CDCl3) δ174.98,151.81,145.03,140.63,139.37,136.64,121.50,116.18,77 .71,47.09,45.38,40.89,34.29,31.67,29.63,29.46,28.34,23.04,22.93,19.39,18.60.

[0131]

[0132] Compound Ⅰ-4-2: white solid, HR-MS m / z: C 27 H 29 NO3,438.2040[M+Na] + (calculated value), 438.2043[M+Na] + (measured value). 1 H NMR (600MHz, CDCl3) δ8.29–8.18(m,2H),7.54(dd,J=5.41.8Hz,3H),7.13(s,1H),5.65(dd,J=4.2,1.8Hz,1H),3. 68(p,J=7.2Hz,1H),3.12(d,J=13.8Hz,1H),2.54(td,J=13.8,4.8Hz,1H),2.33(ddd,J=13.8,6.0,4.2Hz,1H),2.1 5(qt,J=13.8,3.6Hz,1H),1.98(ddd,J=13.8,10.2,1.8Hz,1H),1.83(dt,J=13.8,3.6Hz,1H),1.74(dd,J=10.2,6. 0Hz,1H),1.67–1.60(m,2H),1.39(dd,J=6.6,3.6Hz,6H),1.34(dd,J=13.6,3.6Hz,1H),0.98(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ175.16,162.43,145.70,140.16,136.02,131.51,128.89,127.68,127.14,120.9 0,116.02,77.90,47.16,45.49,40.94,34.33,31.69,29.74,29.34,28.33,23.19,23.07,19.45,18.76.

[0133]

[0134] Compound Ⅰ-4-3: white solid, ESI-MS m / z: 450 [M+H] + , 1H NMR (600MHz, CDCl3) δ8.22(t,J=1.8Hz,1H),8.12(dt,J=7.2,1.2Hz,1H),7.51(dt,J=7.8,1.2Hz,1H),7.47(t,J=7. 8Hz,1H),7.14(s,1H),5.65(dd,J=4.2,1.8Hz,1H),3.66(hept,J=6.6Hz,1H),3.11(d,J=13.8Hz,1H),2.52(td,J=1 3.2,4.8Hz,1H),2.33(ddd,J=13.8,6.0,4.2Hz,1H),2.15(qt,J=13.8,3.6Hz,1H),1.98(ddd,J=13.8,10.2,1.8Hz, 1H), 1.84 (dq, J=10.2, 3.6Hz, 1H), 1.73 (dd, J=10.2, 6.0Hz, 1H), 1.64 (d, J=14.4Hz, 1H), 0.98 (s, 3H), 0.89 (s, 3H). 13 CNMR (150MHz, CDCl3) δ175.02,161.01,145.76,141.34,140.46,136.49,135.02,131.51,130.24,128.84,127.55,1 25.72,121.06,116.30,77.83,47.16,45.45,40.90,34.33,31.69,29.69,29.42,28.34,23.16,23.06,19.45,18.71.

[0135]

[0136] Compound I-4-4: white solid, ESI-MS m / z: 450 [M+H] + , 1H NMR (600MHz, CDCl3) δ8.16 (dd, J=7.2, 1.8Hz, 1H), 7.56 (d, J=7.8Hz, 1H), 7.48–7.40 (m, 2H), 7.16 (s, 1H), 5.67 (d,J=3.0Hz,1H),3.67(hept,J=6.6Hz,1H),3.08(d,J=13.8Hz,1H),2.53(td,J=13.8,4.2Hz,1H),2.40–2.28( m,1H),2.11(ddd,J=16.8,8.4,3.0Hz,1H),2.03–1.94(m,1H),1.78(dt,J=13.8,3.0Hz,1H),1.76–1.72(m,1H) ,1.61(d,J=13.2Hz,1H),1.40(dd,J=7.2,3.6Hz,6H),1.32(td,J=13.2,3.6Hz,1H),0.98(s,3H),0.88(s,3H). 13 C NMR (150MHz, CDCl3) δ175.13,160.56,145.66,140.97,140.49,136.48,132.00,131.37,126.98,126.2 8,116.19,77.84,47.05,45.43,40.94,34.32,31.69,29.72,29.58,28.35,23.12,23.04,19.43,18.71.

[0137]

[0138] Compound Ⅰ-4-5: white solid, ESI-MS m / z: 450 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 8.18 (d, J = 9.0 Hz, 2H), 7.50 (d, J = 8.4 Hz, 2H), 7.13 (s, 1H), 5.65 (d, J = 2.4 Hz, 1H), 3.66 (p, J = 7.2 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.53 (dd, J = 13.8, 4.8 Hz, 1H), 2.37–2.29 (m, 1H), 2.15 (qt, J = 13.8, 3.6 Hz, 1H), 2.03–1.94 (m, 1H), 1.83 (dt, J = 13.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.63 (d, J = 12.0 Hz, 1H), 1.39 (td, J = 7.2, 6.0, 2.4 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.06, 161.44, 145.70, 140.28, 137.73, 129.25, 129.17, 128.91, 125.63, 120.97, 116.21, 77.85, 47.16, 45.46, 40.91, 34.33, 31.69, 29.70, 29.37, 28.35, 23.16, 23.06, 19.44, 18.74.

[0139]

[0140] Compound I-4-6: Yellow solid, ESI-MS m / z: 494 [M + H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.10 (d, J = 8.4 Hz, 2H), 7.66 (d, J = 8.4 Hz, 2H), 7.13 (s, 1H), 5.65 (dd, J = 3.6, 1.8 Hz, 1H), 3.65 (hept, J = 7.2 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.51 (td, J = 13.2, 4.2 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.21–2.07 (m, 1H), 2.02–1.94 (m, 1H), 1.83 (dd, J = 13.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.63 (d, J = 13.2 Hz, 2H), 1.38 (dd, J = 6.6, 3.0 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13C NMR (150MHz, CDCl3) δ175.06,161.51,145.70,141.44,140.31,132.21,129.07,126.20,126.06,120.9 8,116.23,77.85,47.15,45.46,40.91,34.33,31.69,29.69,29.37,28.34,23.16,23.06,19.45,18.74.

[0141]

[0142] Compound Ⅰ-4-7: white solid, ESI-MS m / z: 434 [M+H] + , 1 H NMR (600MHz, CDCl3) δ8.24(dd,J=9.0,5.4Hz,2H),7.22(t,J=8.4Hz,2H),7.13(s,1H),5.65(dd,J=3.6,1 .2Hz,1H),3.66(hept,J=6.6Hz,1H),3.10(d,J=13.8Hz,1H),2.52(td,J=13.2,4.2Hz,1H),2.33(ddd,J=1 3.8,6.0,4.2Hz,1H),2.15(qt,J=13.8,3.6Hz,1H),2.03–1.93(m,1H),1.82(dt,J=13.8,3.6Hz,1H),1.7 3(dd,J=10.2,6.0Hz,1H),1.63(d,J=13.2Hz,1H),1.39(dd,J=7.2,3.0Hz,6H),0.98(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ175.11,165.64,163.97,161.54,145.71,141.49,140.14,136.07,129.92,123.47,120.91,11 6.25,116.13,116.10,77.87,47.15,45.47,40.92,34.33,31.68,29.72,29.36,28.35,23.16,23.05,19.44,18.74.

[0143]

[0144] Compound Ⅰ-4-8: white solid, ESI-MS m / z: 432 [M+H] + , 1H NMR(600MHz, CDCl3)δ11.39(s,1H),8.04–7.95(m,1H),7.52–7.38(m,1H),7.16(s,1H),7.11(d,J=7.8Hz,1H),7 .03(t,J=7.2Hz,1H),5.66(dd,J=3.6,1.2Hz,1H),3.56(hept,J=7.2Hz,1H),3.19–3.06(m,1H),2.62–2.48(m,1H ),2.34(ddd,J=13.8,6.0,4.2Hz,1H),2.16(qt,J=13.8,3.0Hz,1H),1.99(ddd,J=13.8,10.8,1.2Hz,1H),1.84( dq,J=10.2,3.6Hz,1H),1.74(dd,J=10.2,6.0Hz,1H),1.65(s,1H),1.42–1.33(m,6H),0.99(s,3H),0.90(s,3H). 13 C NMR (150MHz, CDCl3) δ174.85,162.37,158.65,144.13,139.20,139.16,136.55,133.76,127.09,121.23,119.73,11 7.40,116.75,110.34,77.75,47.19,45.40,40.88,34.35,31.68,29.77,29.66,28.33,22.87,22.82,19.44,18.72.

[0145]

[0146] Compound I-4-9: white solid, ESI-MS m / z: 510 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 11.48 (s, 1H), 8.06 (d, J = 2.4 Hz, 1H), 7.54 (dd, J = 9.0, 2.4 Hz, 1H), 7.18 (s, 1H), 7.04 (d, J = 9.0 Hz, 1H), 5.67 (dd, J = 3.6, 1.2 Hz, 1H), 3.56 (hept, J = 6.6 Hz, 1H), 3.11 (d, J = 13.8 Hz, 1H), 2.50 (td, J = 13.2, 4.2 Hz, 1H), 2.35 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.16 (qt, J = 13.8, 3.6 Hz, 1H), 2.04–1.94 (m, 1H), 1.89 (dt, J = 13.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.8, 6.0 Hz, 1H), 1.66 (d, J = 14.4 Hz, 1H), 1.41–1.37 (m, 7H), 0.99 (s, 3H), 0.91 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.68, 161.07, 157.71, 144.20, 139.43, 138.88, 137.07, 136.46, 129.10, 121.45, 119.43, 117.03, 111.97, 111.43, 77.67, 47.20, 45.38, 40.83, 34.36, 31.70, 29.74, 28.34, 22.88, 22.82, 19.45, 18.62.

[0147]

[0148] Compound I-4-10: White solid, ESI-MS m / z: 477 [M + H] + , 1H NMR(600MHz, CDCl3)δ7.75(dd,J=8.4,1.8Hz,1H),7.72(d,J=1.8Hz,1H),7.11(s,1H),7.04(d,J=8.4Hz,1H), 6.21(s,1H),5.65(dd,J=3.6,1.2Hz,1H),3.99(s,3H),3.69(hept,J=6.6Hz,1H),3.10(d,J=13.8Hz,1H),2.5 2(td,J=13.8,4.2Hz,1H),2.33(ddd,J=13.8,6.0,4.0Hz,1H),2.14(qt,J=13.8,3.0Hz,1H),2.05–1.95(m,1H ),1.87–1.78(m,1H),1.77–1.70(m,2H),1.63(d,J=13.2Hz,1H),1.43–1.31(m,7H),0.98(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ175.34,162.63,149.03,146.84,145.56,141.64,139.67,135.56,121.86,119.28,115.82,1 14.79,109.87,77.98,56.17,47.15,45.49,40.92,34.33,31.68,29.76,29.09,28.29,23.29,23.11,19.45,18.77.

[0149]

[0150] Compound Ⅰ-4-11: white solid, ESI-MS m / z: 446 [M+H] + , 1H NMR (600MHz, CDCl3) δ8.18(d,J=9.0Hz,2H),7.10(s,1H),7.03(d,J=9.0Hz,2H),5.64(dd,J=3.6,1.2Hz,1H),3.90(s,3H ),3.66(hept,J=6.6Hz,1H),3.10(d,J=13.8Hz,1H),2.53(td,J=13.8,4.2Hz,1H),2.32(ddd,J=13.8,6.0,4.2Hz,1H),2 .14(qt,J=13.2,3.0Hz,1H),2.01–1.94(m,1H),1.82(dt,J=13.8,3.6Hz,1H),1.73(dd,J=10.2,6.0Hz,1H),1.69(d,J=3 .6Hz,1H),1.63(d,J=13.2Hz,1H),1.38(dd,J=6.6,3.6Hz,6H),1.34(dd,J=13.8,3.6Hz,1H),0.98(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ175.27,162.56,162.31,145.56,141.73,139.67,135.49,129.45,120.67,119.68,115.8 3,114.33,77.94,55.47,47.14,45.50,40.94,34.33,31.68,29.77,29.23,28.33,23.20,23.08,19.45,18.77.

[0151]

[0152] Compound Ⅰ-4-12: yellow solid, ESI-MS m / z: 505 [MH] - , 11H NMR (600 MHz, CDCl3) δ 8.52 (s, 1H), 7.98 (s, 1H), 7.16 (s, 1H), 5.71–5.60 (m, 1H), 4.11 (s, 3H), 3.69 (p, J = 6.6 Hz, 1H), 3.12 (d, J = 13.8 Hz, 1H), 2.51 (td, J = 13.2, 4.2 Hz, 1H), 2.40–2.27 (m, 1H), 2.22–2.08 (m, 1H), 2.05–1.93 (m, 1H), 1.86 (d, J = 13.8 Hz, 1H), 1.79–1.62 (m, 3H), 1.39 (dd, J = 7.2, 3.0 Hz, 6H), 0.99 (s, 3H), 0.91 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.05, 160.24, 150.59, 148.70, 145.80, 141.22, 140.44, 136.61, 133.92, 121.05, 118.66, 116.42, 115.66, 115.50, 77.86, 57.09, 47.17, 45.43, 4i.86, 34.34, 31.68, 29.64, 29.20, 28.33, 23.13, 19.44, 18.70.

[0153]

[0154] Compound I-4-13: Yellow solid, ESI-MS m / z: 484 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.36 (d, J = 8.4 Hz, 2H), 7.79 (d, J = 8.4 Hz, 2H), 7.16 (s, 1H), 5.67 (d, J = 2.4 Hz, 1H), 3.68 (hept, J = 7.2 Hz, 1H), 3.12 (d, J = 13.8 Hz, 1H), 2.53 (td, J = 13.8, 4.2 Hz, 1H), 2.38–2.31 (m, 1H), 2.16 (qt, J = 13.8, 3.0 Hz, 1H), i.03–1.9i (m, 1H), 1.84 (dt, J = 13.8, 3.0 Hz, 1H), 1.74 (dd, J = 10'.2, 6.0 Hz, 1H), 1.65 (d, J = 13.2 Hz, 1H), 1.44–1.32 (m, 7H), 0.99 (s, 3H), 0.90 (s, 3H). 13CNMR (150MHz, CDCl3) δ174.99,160.87,145.84,141.34,140.70,136.76,133.08,132.86,130.40,127.93,125.92,124.6 6,122.86,121.15,116.42,77.82,47.17,45.44,40.90,34.34,31.68,29.66,29.43,28.36,23.15,23.05,19.43,18.73.

[0155]

[0156] Compound Ⅰ-4-14: white solid, ESI-MS m / z: 474 [M+H] + , 1 H NMR (600MHz, CDCl3) δ8.32(d,J=8.4Hz,2H),8.20(d,J=8.4Hz,2H),7.15(s,1H),5.69–5.64(m,1H),3.9 8(s,3H),3.68(h,J=6.6Hz,1H),3.12(d,J=13.2Hz,1H),2.54(td,J=13.8,4.2Hz,1H),2.34(dt,J=13.8, 4.2Hz,1H),2.16(q,J=13.8Hz,1H),2.03–1.96(m,1H),1.85(d,J=13.8Hz,1H),1.74(dd,J=10.2,6.0Hz ,1H),1.64(d,J=13.2Hz,1H),1.40(dd,J=6.6,3.0Hz,6H),1.38–1.33(m,1H),0.99(s,3H),0.90(s,3H). 13 C NMR (150MHz, CDCl3) δ175.02,166.36,161.33,145.83,141.45,140.62,136.67,132.51,130.99,130.10,127.54,1 21.10,116.34,77.83,52.45,47.17,45.45,40.90,34.34,31.69,29.67,29.40,28.34,23.18,23.07,19.44,18.73.

[0157]

[0158] Compound Ⅰ-4-15: white solid, ESI-MS m / z: 458 [MH] - ,1 H NMR(600MHz,DMSO-d6)δ8.31(d,J=8.4Hz,2H),8.16(d,J=8.4Hz,2H),7.42(s,1H) ,5.88(d,J=2.4Hz,1H),3.60(hept,J=7.2Hz,1H),2.92(d,J=13.8Hz,1H),2.56(d d,J=13.8,4.2Hz,1H),2.28–2.19(m,1H),2.00–1.88(m,2H),1.75(dd,J=10.2,6. 0Hz, 2H), 1.53 (d, J = 12.6Hz, 1H), 1.37 (d, J = 6.6Hz, 6H), 0.88 (s, 3H), 0.84 (s, 3H). 13 C NMR(150MHz,DMSO-d6)δ174.87,167.07,161.43,145.62,140.99,140.01,137.44,133.93,130.60,127.9 3,121.32,117.26,79.63,77.42,47.06,44.74,34.36,31.61,29.47,28.13,23.34,23.27,19.47,18.73.

[0159]

[0160] Compound I-4-16: yellow solid, ESI-MS m / z: 494 [M+H] + , 1 H NMR(600MHz, CDCl3)δ8.45(d,J=8.4Hz,2H),8.12(d,J=8.4Hz,2H),7.18(s,1H),5.73–5.61( m,1H),3.68(hept,J=6.6Hz,1H),3.14(s,3H),2.53(td,J=13.2,4.2Hz,1H),2.41–2.30(m,1 H),2.16(q,13.8Hz,1H),2.05–1.93(m,1H),1.85(d,J=10.2Hz,1H),1.81–1.69(m,2H),1.65 (d,J=13.8Hz,1H),1.40(dd,J=6.6,2.4Hz,6H),1.38–1.32(m,1H),0.99(s,3H),0.90(s,3H). 13C NMR (150MHz, CDCl3) δ174.96,160.31,145.93,142.67,141.29,140.95,137.13,132.01,128.41,128.05,121.24,1 17.93,116.63,77.80,47.18,45.41,44.47,40.86,34.34,31.68,29.61,29.47,28.37,23.14,23.06,19.43,18.70.

[0161]

[0162] Compound Ⅰ-4-17: white solid, ESI-MS m / z: 473 [M+H] + , 1 H NMR(600MHz, CDCl3)δ8.09(s,1H),7.87(d,J=8.4Hz,2H),7.51(d,J=8.4Hz,2H),7.13(s,1H),5.72–5.66(m,1H ),3.68(hept,J=7.2Hz,1H),3.00(d,J=13.8Hz,1H),2.46(td,J=13.2,4.2Hz,1H),2.37–2.31(m,1H),2.24(s,3 H),2.12(q,J=13.8Hz,1H),2.03–1.96(m,1H),1.84(d,J=13.8Hz,1H),1.74(dd,J=10.2,6.0Hz,1H),1.63(d,J= 13.2Hz,1H),1.49(t,J=7.8Hz,1H),1.42(dd,J=20.4,6.6Hz,6H),1.38–1.32(m,2H),0.99(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ176.33,168.90,161.99,145.45,141.73,141.06,140.05,135.57,128.30,122.11,120.31,1 19.22,115.92,78.28,47.28,45.45,40.84,34.34,31.61,29.69,29.28,28.28,24.69,23.37,22.98,19.49,18.70.

[0163]

[0164] Compound I-4-18: white solid, ESI-MS m / z: 461 [M+H]+ , 1 1H NMR (600 MHz, CDCl3) δ 8.18 (d, J = 9.0 Hz, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.75 (t, J = 7.8 Hz, 1H), 7.69 (t, J = 7.2 Hz, 1H), 7.17 (s, 1H), 5.66 (d, J = 2.4 Hz, 1H), 3.62 (h, J = 6.6 Hz, 1H), 2.97 (d, J = 13.8 Hz, 1H), 2.37–2.30 (m, 2H), 2.09 (qt, J = 13.8, 3.6 Hz, 1H), 2.01–1.94 (m, 1H), 1.84–1.74 (m, 1H), 1.72 (dd, J = 10.2, 6.0 Hz, 1H), 1.61 (d, J = 13.2 Hz, 1H), 1.39 (dd, J = 6.6, 1.8 Hz, 6H), 1.35–1.30 (m, 1H), 0.96 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.89, 158.06, 145.91, 140.84, 137.06, 132.19, 131.27, 124.07, 121.38, 120.98, 116.56, 77.75, 47.06, 45.31, 40.87, 34.31, 31.65, 29.67, 28.07, 23.00, 19.40, 18.63.

[0165]

[0166] Compound I-4-19: White solid, ESI-MS m / z: 461 [M + H] + , 1 1H NMR (600 MHz, CDCl3) δ 9.07 (s, 1H), 8.55 (d, J = 7.8 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 7.75 (t, J = 7.13 C NMR (150MHz, CDCl3) δ174.89,159.97,148.72,145.89,141.19,140.89,137.04,133.03,130.16,128.92,125 .79,122.53,121.27,116.64,77.79,47.18,45.43,40.88,34.35,31.69,29.49,28.38,23.15,19.44,18.69.

[0167]

[0168] Compound Ⅰ-4-20: white solid, ESI-MS m / z: 461 [M+H] + , 1 H NMR (600MHz, CDCl3) δ8.44–8.36(m,4H),7.18(s,1H),5.68(dd,J=3.6,1.2Hz,1H),3.67(hept ,J=7.2Hz,1H),3.13(d,J=13.8Hz,1H),2.53(td,J=13.2,4.2Hz,1H),2.36(ddd,J=13.8,6.0,4 .2Hz,1H),2.16(qt,J=13.8,3.6Hz,1H),2.06–1.94(m,1H),1.85(dq,J=13.8,3.6Hz,1H),1.74 (dd,J=10.2,6.0Hz,1H),1.65(d,J=11.4Hz,1H),1.44–1.39(m,7H),0.99(s,3H),0.91(s,3H). 13 CNMR (150MHz, CDCl3) δ174.85,160.02,149.37,146.00,141.33,141.06,137.30,132.70,128.42,12 4.20,121.30,116.71,77.77,47.18,45.43,40.88,34.35,31.69,29.51,28.39,23.12,19.43,18.71.

[0169]

[0170] Compound Ⅰ-4-21: white solid, ESI-MS m / z: 406 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 7.68 (dd, J = 1.8, 0.6 Hz, 1H), 7.29–7.27 (m, 1H), 7.15 (s, 1H), 6.62 (dd, J = 3.6, 1.8 Hz, 1H), 5.65 (dd, J = 4.2, 1.8 Hz, 1H), 3.72 (hept, J = 7.2 Hz, 1H), 3.06 (d, J = 14.4 Hz, 1H), 2.48 (td, J = 13.8, 4.2 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.13 (qt, J = 13.8, 3.0 Hz, 1H), 1.98 (ddd, J = 13.8, 10.2, 1.2 Hz, 1H), 1.87–1.76 (m, 1H), 1.72 (dd, J = 10.2, 6.0 Hz, 1H), 1.66–1.56 (m, 1H), 1.36 (dd, J = 7.2, 4.2 Hz, 6H), 0.97 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.01, 154.79, 145.85, 145.03, 142.46, 141.01, 140.29, 136.32, 120.91, 116.21, 114.47, 112.21, 77.8, 47.11, 45.42, 40.87, 34.31, 31.67, 29.67, 28.23, 23.25, 2(.09, 19.42, 18.67.

[0171]

[0172] Compound I-4-22: ESI-MS m / z: 478 [M+H] + , 1 It should be noted that in the translation of the carbon nuclear magnetic resonance data in , there may be a small error in the value "77.81" in the original Chinese text which is translated as "77. (There is a lack of a complete digit after the decimal point)". It is recommended to check the original data for accuracy.1H NMR (600 MHz, CDCl3) δ 7.24 (d, J = 3.6 Hz, 1H), 7.15 (s, 1H), 6.63 (d, J = 3.0 Hz, 1H), 5.65 (d, J = 2.4 Hz, 1H), 5.19 (s, 2H), 3.74 (hept, J = 7.2 Hz, 1H), 3.05 (d, J = 13.8 Hz, 1H), 2.46 (td, J = 13.8, 4.2 Hz, 1H), 2.39–2.29 (m, 1H), 2.13 (s, 3H), 2.02–1.95 (m, 1H), 1.80 (dt, J = 13.8, 3.6 Hz, 1H), 1.72 (dd, J = 10.8, 6.0 Hz, 1H), 1.63 (d, J = 12.6 Hz, 1H), 1.36 (dd, J = 7.2, 4.2 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.98, 170.50, 154.43, 153.15, 145.01, 142.73, 140.96, 140.42, 136.51, 120.91, 116.26, 115.35, 112.85, 77.80, 57.88, 47.11, 45.40, 40.86, 31.66, 29.64, 28.79, 28.23, 23.30, 23.12, 19.42, 18.66.

[0173]

[0174] Compound I-4-23: Yellow solid, ESI-MS m / z: 420 [M + H] + , 1 1H NMR (600 MHz, CDCl3) δ 7.18 (d, J = 3.6 Hz, 1H), 7.13 (s, 1H), 6.22 (d, J = 3.0 Hz, 1H), 5.64 (d, J = 2.4 Hz, 1H), 3.75 (hept, J = 7.2 Hz, 1H), 3.04 (d, J = 13.8 Hz, 1H), 2.48 (s, 3H), 2.32 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.12 (qt, J = 13.8, 3.0 Hz, 1H), 1.98 (dd, J = 14.4, 11.4 Hz, 1H), 1.79 (dt, J = 13.8, 3.6 Hz, 1H), 1.72 (dd, J = 10.8, 6.0 Hz, 1H), 1.62 (d, J = 12.6 Hz, 1H), 1.35 (dd, J = 6.6, 4.8 Hz, 6H), 0.97 (s, 3H), 0.89 (s, 3H). 13C NMR (150MHz, CDCl3) δ175.10,156.86,155.09,144.89,141.15,140.81,140.03,135.98,120.70,116.01,1 08.70,77.85,47.10,45.43,40.88,34.32,31.67,29.70,28.64,28.22,23.35,23.16,19.43,18.68,14.11.

[0175]

[0176] Compound I-4-24: yellow solid, ESI-MS m / z: 422 [M+H] + , 1 H NMR(600MHz, CDCl3)δ7.90(d,J=4.8Hz,1H),7.55(d,J=6.0Hz,1H),7.22–7.16(m,1H),7.12(s,1H),5.64(d d,J=3.6,1.2Hz,1H),3.66(hept,J=7.2Hz,1H),3.07(d,J=13.8Hz,1H),2.49(td,J=13.8,4.2Hz,1H),2.32( ddd,J=13.8,6.0,4.2Hz,1H),2.13(qt,J=13.8,3.6Hz,1H),2.04–1.90(m,1H),1.80(dq,J=10.8,3.6Hz,1H) ,1.73(dd,J=10.2,6.0Hz,1H),1.62(dd,J=13.2,1.2Hz,1H),1.40–1.31(m,,7H),0.98(s,3H),0.89(s,3H). 13 C NMR (150MHz, CDCl3) δ175.12,158.40,145.31,141.48,139.95,136.03,130.22,130.01,129.58,128.20,12 0.74,116.11,77.87,47.12,45.45,40.91,34.33,31.68,29.71,29.14,28.25,23.21,23.07,19.45,18.73.

[0177]

[0178] Compound Ⅰ-4-25: yellow solid, ESI-MS m / z: 423 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 8.11 (d, J = 3.0 Hz, 1H), 7.63 (d, J = 3.0 Hz, 1H), 7.19 (s, 1H), 5.67 (d, J = 2.4 Hz, 1H), 3.72 (hept, J = 7.2 Hz, 1H), 3.11 (d, J = 13.2 Hz, 1H), 2.55 (td, J = 13.8, 4.8 Hz, 1H), 2.34 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.16–2.07 (m, 1H), 2.02–1.94 (m, 1H), 1.84 (d, J = 13.8 Hz, 1H), 1.73 (dd, J = 10.8, 6.0 Hz, 1H), 1.61 (d, J = 13.2 Hz, 1H), 1.38 (dd, J = 7.2, 3.0 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.84, 156.51, 154.68, 145.71, 145.26, 141.18, 140.85, 137.54, 123.01, 121.49, 116.77, 77.72, 47.18, 45.37, 40.78, 34.32, 31.70, 29.58, 29.18, 28.15, 23.23, 23.08, 19.44, 18.58.

[0179]

[0180] Compound I-4-26: Purple solid, ESI-MS m / z: 417 [M + H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.85 (d, J = 4.8 Hz, 1H), 8.29 (d, J = 7.8 Hz, 1H), 7.97–7.87 (m, 1H), 7.47 (dd, J = 7.2, 4.8 Hz, 1H), 7.19 (s, 1H), 5.67 (d, J = 2.4 Hz, 1H), 3.84 (h, J = 7.2 Hz, 1H), 3.14 (d, J = 13.8 Hz, 1H), 2.34 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.14 (qt, J = 13.8, 3.6 Hz, 1H), 2.04–1.95 (m, 1H), 1.77–1.72 (m, 1H), 1.62 (d, J = 12.6 Hz, 1H), 1.37 (m, 7H), 0.98 (s, 3H), 0.89 (s, 3H). 13C NMR (150MHz, CDCl3) δ175.03,161.02,150.62,145.93,145.89,141.27,141.23,137.12,137.04,125.57,123.5 9,121.20,116.34,77.83,47.19,45.40,40.82,34.33,31.70,29.63,28.79,28.24,23.39,23.20,19.44,18.68.

[0181]

[0182] Compound I-4-27: yellow oil, ESI-MS m / z: 438 [M+H] + , 1 H NMR (600MHz, CDCl3) δ7.08(s,1H),5.63(dd,J=3.6,1.2Hz,1H),3.58(hept,J=7.2Hz,1H),2.97(d,J =14.4Hz,1H),2.93(t,J=7.8Hz,2H),2.39(td,J=13.8,4.8Hz,1H),2.30(ddd,J=13.8,6.0,4.0Hz,1H ),2.09(qt,J=13.8,3.0Hz,1H),1.98–1.91(m,1H),1.86(p,J=7.8Hz,2H),1.69(dd,J=10.8,6.0Hz, 1H), 1.60 (d, J = 14.4Hz, 1H), 1.46–1.41 (m, 2H), 1.38–1.23 (m, 20H), 0.96 (s, 3H), 0.90–0.87 (m, 6H). 13 C NMR (150MHz, CDCl3) δ175.26,166.83,145.72,140.70,139.39,135.33,120.66,116.17,115.42,77.87,47.06,45.41,40.9 2,34.29,31.81,31.66,29.74,29.21,29.16,29.15,29.08,28.76,28.35,27.12,23.13,23.01,22.65,19.40,18.64,14.11.

[0183]

[0184] Compound I-4-28: yellow oil, ESI-MS m / z: 396 [M+H] + , 1H NMR (600MHz, CDCl3) δ7.08(s,1H),5.63(dd,J=4.2,1.8Hz,1H),3.58(hept,J=7.2Hz,1H),2.99–2.92( m,3H),2.78–2.73(m,1H),2.54(dt,J=27.0,7.8Hz,1H),2.39(td,J=13.8,4.8Hz,1H),2.30(ddd,J=13. 8,6.0,4.2Hz,1H),2.14–2.05(m,1H),1.98–1.92(m,1H),1.87–1.83(m,2H),1.79–1.73(m,1H),1.71–1 .65(m,2H),1.64–1.58(m,2H),1.51–1.41(m,3H),1.35–1.30(m,7H),1.01–0.93(m,10H),0.88(s,3H). 13 C NMR (150MHz, CDCl3) δ175.29,166.81,157.55,146.54,145.72,140.69,139.40,136.92,135.33,120.65,115.43,77. 88,47.06,45.40,40.92,34.29,31.66,29.73,29.17,29.07,28.50,28.34,23.13,23.01,22.37,19.40,18.63,13.71.

[0185]

[0186] Compound I-4-29: white solid, ESI-MS m / z: 481 [M+Na] + . 11H NMR (600 MHz, CDCl3) δ 8.09 (d, J = 9.0 Hz, 2H), 7.07 (s, 1H), 6.77 (d, J = 9.0 Hz, 2H), 5.63 (dd, J = 3.6, 1.2 Hz, 1H), 3.67 (hept, J = 7.2 Hz, 1H), 3.08 (s, 6H), 2.54 (td, J = 14.4, 4.2 Hz, 1H), 2.31 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.14 (qt, J = 13.8, 3.0 Hz, 1H), 1.98 (ddd, J = 13.8, 10.8, 1.4 Hz, 1H), 1.81 (dt, J = 13.8, 3.6 Hz, 1H), 1.74 (dd, J = 10.8, 6.0 Hz, 1H), 1.37 (dd, J = 6.6, 3.6 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.43, 163.59, 152.37, 145.40, 142.08, 139.01, 134.76, 129.14, 120.33, 115.48, 114.20, 111.54, 78.03, 47.13, 45.53, 40.97, 40.15, 34.32, 31.69, 29.85, 29.07, 28.31, 23.25, 23.11, 19.46, 18.81.

[0187] Compound I-4-30: White solid, ESI-MS m / z: 481 [M+Na] + , 1 H NMR (600 MHz, CDCl3) δ 8.11 (d, J = 9.0 Hz, 2H), 7.00 (s, 1H), 6.77 (d, J = 9.0 Hz, 2H), 5.59 (dd, J = 3.6, 1.2 Hz, 1H), 3.40 (hept, J = 7.2 Hz, 1H), 3.08 (s, 6H), 2.91 (td, J = 13.8, 4.2 Hz, 1H), 2.29 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.08 (qt, J = 13.8, 3.6 Hz, 1H), 1.94 (ddd, J = 13.8, 10.2, 1.2 Hz, 1H), 1.82 (dt, J = 13.8, 3.6 Hz, 1H), 1.72 (dd, J = 10.2, 6.0 Hz, 1H), 1.40 (dd, J = 7.2, 1.2 Hz, 6H), 0.97 (s, 3H), 0.87 (s, 3H). 13C NMR (150MHz, CDCl3) δ175.84,163.84,152.40,149.31,138.97,134.90,129.83,129.17,128.09,115.75,114.4 0,111.52,77.77,48.44,45.47,40.98,40.15,34.36,31.79,29.88,29.28,28.65,22.60,22.54,19.54,18.78.

[0188]

[0189] Compound I-4-31: white solid, ESI-MS m / z: 416 [M+H] + , 1 H NMR(600MHz,Chloroform-d)δ8.29–8.18(m,2H),7.54(dd,J=5.2,1.9Hz,3H),7.13(s,1H),5.65(dd,J=3.9,1.6Hz, 1H),3.68(p,J=6.9Hz,1H),3.12(d,J=13.6Hz,1H),2.54(td,J=13.5,4.5Hz,1H),2.33(ddd,J=13.8,6.0,4.0Hz,1H) ,2.15(qt,J=13.8,3.4Hz,1H),1.98(ddd,J=13.8,10.5,1.6Hz,1H),1.83(dt,J=14.0,3.8Hz,1H),1.74(dd,J=10.5, 6.0Hz,1H),1.67–1.60(m,2H),1.39(dd,J=6.9,3.3Hz,6H),1.34(dd,J=13.6,3.5Hz,1H),0.98(s,3H),0.89(s,3H). 13 CNMR(150MHz,Chloroform-d)δ175.16,145.70,140.16,136.02,131.51,128.89,127.68,127.14,1 16.02,77.90,47.16,45.49,40.94,34.33,31.69,29.74,29.34,28.33,23.19,23.07,19.45,18.76.

[0190] Example 2: Synthesis of benzene-substituted carnosol derivatives

[0191]

[0192] Step 1: Preparation of I-2

[0193] I-1 (10.0 g, 30.5 mmol) and 100 mL of dry 1,4-dioxene were added to a 250 mL round-bottom flask and stirred at room temperature until completely dissolved. Dichlorodicyanobenzoquinone (DDQ) (7.61 g, 33.5 mmol) was slowly added and stirred at room temperature for 0.5 h. The reaction was complete by TLC. The reaction mixture was filtered under reduced pressure, and the filter cake was washed with 50 mL of dichloromethane. 20 g of column chromatography silica gel was added to the filtrate and the mixture was evaporated under reduced pressure until solvent-free. The residue was ground into a powder and poured into a Buchner funnel. The powder was washed with 200 mL of dichloromethane:ethyl acetate (50:1). The filtrate was evaporated to dryness under reduced pressure. 30 mL of anhydrous ethanol was added to the residue and heated under reflux to completely dissolve the solid. The mixture was then slowly cooled and crystallized. After filtration, I-2 (7.7 g, 77.5% yield) was obtained. 1 H-NMR (600MHz, Acetone-d6) δ6.77 (s, 1H), 5.43 (s, 1H), 3.30 (dq, J = 13.6, 6.8Hz, 1H), 2.85-2. 79(m,1H),2.54(td,J=13.8,4.4Hz,1H),2.25-2.16(m,1H),1.96(dt,J=13.7,3.2Hz,1H),1.85( ddd,J=13.8,10.7,1.5Hz,1H),1.69(dd,J=10.7,5.8Hz,1H),1.57(dq,J=10.5,3.7Hz,1H),1.54 -1.47(m,1H),1.32(td,J=13.4,3.2Hz,1H),1.19(dd,J=6.8,3.9Hz,6H),0.88(d,J=3.5Hz,6H).

[0194] Step 2: Preparation of I-3-2

[0195]

[0196] Under nitrogen, compound I-2 (800 mg, 2.42 mmol) and dry dichloromethane (30 mL) were added to a 100 mL round-bottom flask and stirred until completely dissolved. Pyridine (780 μL, 764 mg, 9.68 mmol) was then added, the temperature was lowered to 0°C, and the mixture was stirred for 10 min. Trifluoromethanesulfonic anhydride (900 μL, 1509 mg, 5.35 mmol) was then slowly added dropwise. The mixture was stirred for 10 min, warmed to room temperature, and the reaction was continued for 2 h. Completion was monitored by TLC. Dilute hydrochloric acid (30 mL, 0.1 M) was slowly added, the layers separated, and the organic phase was washed with saturated sodium bicarbonate solution (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, volume ratio 12:1) to obtain 900 mg of a white solid (I-3-2) in a yield of 62.6%. ESI-MS m / z:595[M+Na] + , 1 H NMR (600MHz, DMSO-d6) δ7.86(s,1H),5.83(s,1H),3.10(hept,J=6.6Hz,1H),2.74(s,1H),2.22–2.13(m,1H),1.98(t,J=10.2Hz,1H),1.94–1. 84(m,1H),1.75(qt,J=13.8,3.0Hz,1H),1.63–1.52(m,2H),1.40(d,J=13.2Hz,1H),1.23(d,J=6.6Hz,4H),1.14(d,J=6.6Hz,3H),0.77(s,6H). 13 C NMR(150MHz,DMSO-d6)δ172.52,144.08,142.54,137.13,134.87,124.01,119.49 ,117.37,76.28,49.00,44.25,34.95,31.48,28.37,28.11,27.61,19.68,18.60.

[0197] Step 3: Preparation of I-5

[0198]

[0199] Synthesis of compounds I-5-1 to I-5-5: Under nitrogen, compound 35 (200 mg, 0.33 mmol), the corresponding boronic acid derivative (1.32 mmol), potassium carbonate (140 mg, 1.01 mmol), tetrakistriphenylphosphine palladium (20 mg, 0.017 mmol), and dry toluene (10 mL) were added to a 25 mL round-bottom flask. The mixture was heated to 90°C for 6-20 h. The reaction was monitored for completion by TLC. After cooling to room temperature, saturated sodium bicarbonate solution (20 mL) was added, stirred for 5 minutes, and the layers were separated. The aqueous phase was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography to obtain compounds I-5-1 to I-5-5.

[0200]

[0201] Compound Ⅰ-5-1: white solid, ESI-MS m / z: 375 [M+H] + . 1 H NMR(600MHz, Methanol-d4)δ7.32–7.26(m,3H),7.27–7.21(m,1H),7.12(dd,J=8.4,1.2Hz,2H),6.92(s,1H),5.5 7(dd,J=4.2,1.2Hz,1H),2.92(hept,J=6.6Hz,1H),2.33(d,J=13.8Hz,1H),2.19(ddd,J=13.8,6.0,4.2Hz,1H),1 .95–1.87(m,1H),1.84(ddd,J=13.8,10.8,1.8Hz,1H),1.71(td,J=13.2,4.8Hz,1H),1.55(ddt,J=13.8,7.2,3.6 Hz,1H),1.43(dt,J=10.8,4.8Hz,2H),1.18–1.10(m,2H),1.03(dd,J=6.6,2.4Hz,6H),0.82(s,3H),0.77(s,3H). 13 C NMR (150MHz, CDCl3) δ176.07,145.31,141.76,141.21,137.70,137.41,129.22,128.09,126.98,123.5 7,119.86,77.47,46.86,45.51,41.02,34.35,31.51,29.62,29.32,26.68,24.41,24.16,19.37,18.50.

[0202]

[0203] Compound Ⅰ-5-2: yellow solid, ESI-MS m / z: 393 [M+H] + , 1 H NMR (600MHz, CDCl3) δ7.26 (s, 1H), 7.20 (dd, J = 8.4, 5.4Hz, 2H), 7.10 (t, J = 8.4Hz, 2H), 7.02 (s, 1H), 5.59 (d, J =2.4Hz,1H),2.98(p,J=6.6Hz,1H),2.57(d,J=13.2Hz,1H),2.31(ddd,J=13.8,6.0,4.2Hz,1H),2.07(qt,J=1 3.8,3.6Hz,1H),1.99(ddd,J=13.8,10.8,1.8Hz,1H),1.81(td,J=13.2,4.2Hz,1H),1.69(dt,J=13.8,3.6Hz, 1H),1.62–1.52(m,2H),1.22(td,J=13.2,3.6Hz,1H),1.13(dd,J=25.2,6.6Hz,6H),0.96(s,3H),0.87(s,3H). 13 C NMR (150MHz, CDCl3) δ175.98,162.84,161.21,145.44,140.13,137.81,137.62,130.76,130.71,123.61,11 9.93,114.95,77.41,46.84,45.49,41.00,34.35,31.50,29.58,29.33,26.67,24.34,24.09,19.36,18.47.

[0204]

[0205] Compound Ⅰ-5-3: yellow solid, ESI-MS m / z: 409 [M+H] + , 1H NMR (600MHz, CDCl3) δ7.38(d,J=8.4Hz,2H),7.26(s,1H),7.18(d,J=8.4Hz,2H),7.01(s,1H),5.59(d, J=2.4Hz,1H),2.97(hept,J=7.2Hz,1H),2.57(d,J=13.2Hz,1H),2.35–2.28(m,1H),2.07(qt,J=13.8,3 .6Hz,1H),1.98(ddd,J=13.8,10.8,1.8Hz,1H),1.80(td,J=13.2,4.8Hz,1H),1.69(ddd,J=13.8,6.6, 3.6Hz,1H),1.25–1.18(m,2H),1.15(d,J=6.6Hz,3H),1.11(d,J=6.6Hz,3H),0.96(s,3H),0.87(s,3H). 13 C NMR (150MHz, CDCl3) δ175.93,145.34,140.12,139.91,137.90,137.77,133.11,130.54,128.31,12 3.43,119.98,46.84,45.47,40.98,34.35,31.50,29.56,29.35,26.66,24.34,24.10,19.36,18.46.

[0206]

[0207] Compound I-5-4: yellow solid, ESI-MS m / z: 389 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 7.25 (s, 1H), 7.22 (d, J = 7.8 Hz, 2H), 7.14 (d, J = 7.8 Hz, 2H), 7.04 (s, 1H), 5.59 (d, J = 2.4 Hz, 1H), 3.04 (hept, J = 6.6 Hz, 1H), 2.56 (d, J = 13.2 Hz, 1H), 2.41 (s, 3H), 2.30 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.07 (qt, J = 13.8, 3.0 Hz, 1H), 1.98 (ddd, J = 13.8, 10.8, 1.2, 1H), 1.80 (td, J = 13.2, 4.8 Hz, 1H), 1.68 (dt, J = 13.8, 3.6 Hz, 2H), 1.57 (dd, J = 10.8, 6.0 Hz, 2H), 1.21 (td, J = 13.8, 3.6 Hz, 1H), 1.13 (dd, J = 22.2, 6.6 Hz, 6H), 0.95 (s, 3H), 0.87 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 176.07, 145.36, 141.14, 138.80, 137.62, 137.23, 136.62, 129.07, 128.77, 123.64, 119.80, 77.46, 46.82, 45.48, 41.00, 34.32, 31.49, 29.61, 29.29, 26.64, 24.41, 24.17, 21.18, 19.35, 18.48.

[0208]

[0209] Compound I-5-5: ESI-MS m / z: 405 [M + H] + , 1H NMR (600MHz, CDCl3) δ7.24(s,1H),7.17(d,J=8.4Hz,2H),7.04(s,1H),6.95(d,J=8.4Hz,2H),5.5 8(d,J=2.4Hz,1H),3.86(s,3H),3.06(hept,J=6.6Hz,1H),2.57(d,J=12.6Hz,1H),2.35–2.25(m, 1H),2.07(qt,J=13.6,3.6Hz,1H),2.02–1.95(m,1H),1.68(dt,J=13.8,3.6Hz,1H),1.62–1.54(m ,2H),1.31–1.19(m,3H),1.15(d,J=6.6Hz,3H),1.11(d,J=6.6Hz,3H),0.95(s,3H),0.87(s,3H). 13 C NMR (150MHz, CDCl3) δ176.11,158.67,145.52,140.84,137.65,137.20,134.08,130.26,123.77,119.84,1 13.52,77.48,55.31,46.84,45.50,41.02,34.34,31.51,29.63,29.29,26.67,24.42,24.17,19.37,18.50.

[0210] Example 3: Synthesis of 7-20-lactam carnosol

[0211]

[0212] Step 1: Synthesis of Compound II-1

[0213] Under nitrogen, carnosic acid (I-1, 5.0 g, 15.1 mmol), 2,2-dimethoxypropane (6.2 g, 60.0 mmol), pyridinium p-toluenesulfonate (188 mg, 0.75 mmol), and chloroform (50 mL) were added to a 150 mL round-bottom flask. The mixture was stirred to dissolve the solid, heated to 70°C, and refluxed for 20 h. The reaction was monitored for completion by TLC. After cooling to room temperature, saturated sodium bicarbonate solution (50 mL) was added and stirred for 10 min. The layers were separated, and the aqueous phase was extracted with dichloromethane (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, volume ratio 15:1) to obtain 5.54 g of (II-1) as a white solid in a 98.9% yield. ESI-MS m / z: 371 [MH] - , 1H NMR(600MHz, CDCl3) δ6.42(s,1H),3.29(d,J=13.2Hz,1H),2.93–2.87(m,1H),2.85(dd,J=15.6,4.2Hz,1H) ,2.77(ddd,J=16.8,12.0,6.6Hz,1H),2.35(qd,J=12.6,6.0Hz,1H),2.06(ddd,J=17.4,13.8,10.2Hz,1H), 1.81(dd,J=13.2,6.6Hz,1H),1.61(s,3H),1.54(d,J=13.2Hz,2H),1.46(d,J=13.2Hz,1H),1.33(s,3H),1. 27(td,J=13.2,4.2Hz,1H),1.20(t,J=6.6Hz,6H),1.15(dd,J=13.8,3.6Hz,1H),0.95(s,3H),0.83(s,3H). 13 C NMR (150MHz, CDCl3) δ180.97,144.75,142.57,130.01,128.55,120.49,119.10,116.47,53 .44,46.77,41.89,34.06,32.39,30.79,28.38,25.65,25.34,22.24,21.98,19.97,18.70.

[0214] Step 2: Synthesis of Compound II-2

[0215] Under nitrogen, compound II-1 (5.5 g, 14.8 mmol), potassium carbonate (3.05 g, 22.1 mmol), and acetone (60 mL) were added to a 150 mL round-bottom flask and stirred until homogeneously suspended. Methyl iodide (3.15 g, 22.1 mmol) was then added dropwise. The mixture was heated to 65°C and refluxed for 10 h. Completion of the reaction was monitored by TLC. After cooling to room temperature, the reaction mixture was filtered under reduced pressure to remove the potassium carbonate. The filter cake was washed with dichloromethane (20 mL). The filtrate was evaporated to dryness under reduced pressure, and saturated ammonium chloride solution (50 mL) was added. The mixture was then extracted with dichloromethane (50 mL x 3). The extract was dried over anhydrous sodium sulfate, filtered, and the solvent evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, volume ratio 30:1) to obtain 5.5 g of (II-2) as a colorless oil in a yield of 96.5%. 1H NMR (600MHz, CDCl3) δ6.43 (s, 1H), 3.59 (s, 3H), 3.36 (d, J = 13.2Hz, 1H), 2.95–2.85 (m, 2H) ,2.79(ddd,J=17.4,12.0,6.0Hz,1H),2.38(qd,J=12.6,6.0Hz,1H),2.05(qt,J=13.8,4.2H z,1H),1.82(dd,J=13.2,6.6Hz,1H),1.65(s,3H),1.58–1.55(m,1H),1.54(s,3H),1.46(d ,J=13.2Hz,1H),1.33–1.24(m,1H),1.21(dd,J=7.2,3.6Hz,6H),0.96(s,3H),0.76(s,3H).

[0216] Step 3: Synthesis of Compound II-3

[0217] Compound II-2 (5.5 g, 14.7 mmol) was added to acetic acid (30 mL) and stirred to dissolve. Chromium trioxide (3.2 g, 32.5 mmol) was then added and allowed to react at room temperature for 8 h. The reaction was complete by TLC. The acetic acid was evaporated under reduced pressure, and ethyl acetate (50 mL) and saturated sodium bicarbonate solution (50 mL) were added. After stirring for 10 min, the mixture was separated and the aqueous phase was extracted with ethyl acetate (50 mL x 2). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phases were filtered and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, volume ratio 30:1) to obtain 3.8 g of a colorless oil (II-3) in a yield of 66.7%. 1 H NMR (600MHz, CDCl3) δ7.63(s,1H),3.61(s,3H),3.51(d,J=13.8Hz,1H),3.19(dd,J= 18.0,14.4Hz,1H),2.99(hept,J=6.6Hz,1H),2.65(dd,J=18.0,4.8Hz,1H),2.12(dd ,J=14.4,4.8Hz,1H),1.99(qt,J=13.8,3.6Hz,1H),1.66(d,J=23.4Hz,6H),1.50(d, J=13.2Hz,1H),1.34(m,2H),1.24(dd,J=10.8,7.2Hz,6H),0.94(s,3H),0.82(s,3H).

[0218] Step 4: Synthesis of Compound II-4

[0219]

[0220] Synthesis of compound II-4-1: Under nitrogen protection, compound II-3 (2.0 g, 5.2 mmol), hydroxylamine hydrochloride (722 mg, 10.4 mmol), sodium acetate (853 mg, 10.4 mmol) and anhydrous ethanol (50 mL) were added to a 100 mL round-bottom flask, stirred and dissolved, heated to 85 ° C, refluxed for 12 h, TLC monitored the disappearance of compound II-3, cooled to room temperature, evaporated ethanol under reduced pressure, added saturated sodium bicarbonate solution (50 mL), extracted with ethyl acetate (50 mL × 3), and then added to The extract was dried over anhydrous sodium sulfate, filtered, and the solvent evaporated under reduced pressure. Under nitrogen, the residue was added to a 250 mL flask, followed by ammonium acetate (4.0 g, 52.0 mmol), sodium cyanoborohydride (1.6 g, 26.0 mmol), and methanol (80 mL). The mixture was cooled to 0°C and stirred for 10 min. Titanium trichloride solution (15% by mass, 6.4 mL, containing 960 mg of titanium trichloride, 6.2 mmol) was added. The mixture was allowed to react at 0°C for 1 h, then allowed to return to room temperature and continue to react for 6 h. Completion was monitored by TLC. The solvent was evaporated under reduced pressure, and saturated sodium bicarbonate solution (50 mL) was added. The mixture was extracted with ethyl acetate (50 mL × 3). The extract was dried over anhydrous sodium sulfate, filtered, and the solvent evaporated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane-methanol, volume ratio 30:1) to obtain 1.81 g of a colorless oil (II-4-1) in an 87.9% yield. ESI-MS m / z:402[M+H] + , 1 H NMR (600MHz, CDCl3) δ6.80(s,1H),4.64(dd,J=9.6,5.4Hz,1H),3.73(s,3H),3.43(d,J=13.8Hz,1H),2.96(hept,J=7.2Hz,1H),2.56(ddd,J=14.4,9.6,4 .8Hz,1H),2.14(td,J=13.8,6.0Hz,1H),1.93(m,1H),1.70–1.58(m,8H),1.4 8(d,J=13.2Hz,1H),1.23(dd,J=12.0,7.2Hz,6H),0.96(s,3H),0.77(s,3H). 13 CNMR (150MHz, CDCl3) δ176.77,145.44,144.62,130.30,124.96,122.44,120.90,117.62,52.89,5 1.10,50.70,46.95,41.42,33.82,33.50,31.54,28.50,25.78,25.11,21.88,21.79,19.88,19.48.

[0221]

[0222] Synthesis of compounds II-4-2 to II-4-9: Under nitrogen protection, compound II-3 (200 mg, 0.52 mmol), the corresponding amine (1.0 mmol) and 1,4-dioxane (8 mL) were added to a 50 mL round-bottom flask and stirred until dissolved. Tetraisopropyl titanate (284 mg, 1.0 mmol) was then added and heated to 110 ° C. The reaction was refluxed for about 12 h. The disappearance of compound II-3 was monitored by TLC. The mixture was cooled to room temperature, anhydrous ethanol (10 mL) was added to the reaction flask, and sodium borohydride (190 mg, 5.2 mmol) was slowly added. The reaction was carried out at room temperature for about 5 h. The reaction was completed by TLC detection. Water (1 mL) was added to quench the reaction, at which time a large amount of white solid appeared. The reaction solution was filtered, and the filter cake was washed with dichloromethane (20 mL). The filtrate was collected and the solvent was evaporated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with ethyl acetate (20 mL×3). Anhydrous sodium sulfate was added to the extract and dried. After filtration, the solvent was evaporated and the residue was purified by silica gel column chromatography.

[0223]

[0224] Compound II-4-2: colorless oil, ESI-MS m / z: 492 [M+H] + , 1 H NMR (600MHz, CDCl3) δ7.41(d,J=7.2Hz,2H),7.33(t,J=7.8Hz,2H),7.24(t,J=7.2Hz,1H),7.11(s,1H),3.95 (dd,J=10.8,6.7Hz,1H),3.80(q,J=13.2Hz,2H),3.61(s,3H),3.38(d,J=13.2Hz,1H),2.96(hept,J=7.2Hz, 1H),2.57–2.40(m,1H),2.20–2.12(m,1H),2.06(qt,J=13.8,3.6Hz,1H),1.64(s,3H),1.62–1.53(m,5H),1. 47(d,J=13.2Hz,1H),1.24(dd,J=13.8,6.6Hz,6H),1.15(td,J=13.2,4.2Hz,1H),0.97(s,3H),0.80(s,3H). 13C NMR (150MHz, CDCl3) δ175.09,144.32,143.23,141.35,132.30,128.31,126.74,122.25,118.06,116.53,57.1 0,52.00,51.51,48.80,47.26,41.69,34.10,33.79,32.30,28.66,25.68,25.48,22.28,22.05,20.07,19.90.

[0225]

[0226] Compound II-4-3: colorless oil, 1 H NMR (600MHz, CDCl3) δ7.35(d,J=8.4Hz,2H),7.29(d,J=8.4Hz,2H),7.09(s,1H),3.93(dd,J=10.8,6.6Hz,1 H),3.77(q,J=13.2Hz,2H),3.62(s,3H),3.38(d,J=13.2Hz,1H),2.95(hept,J=7.2Hz,1H),2.52–2.43(m,1H ),2.14(dd,J=12.6,6.6Hz,1H),2.05(dddt,J=17.4,13.8,8.4,3.6Hz,1H),1.64(s,3H),1.57(q,J=6.0,5.4 Hz,5H),1.47(d,J=13.2Hz,1H),1.27–1.20(m,8H),1.15(td,J=13.2,4.2Hz,1H),0.97(s,3H),0.79(s,3H).

[0227]

[0228] Compound II-4-4: colorless oil, 1H NMR (600MHz, CDCl3) δ7.54–7.41(m,4H),7.04(s,1H),3.87(dd,J=10.2,6.6Hz,1H),3.79(q,J=13 .2Hz,2H),3.55(s,3H),3.32(d,J=13.2Hz,1H),2.89(hept,J=7.2Hz,1H),2.48–2.36(m,1H),2.07 (ddd,J=12.6,6.6,1.8Hz,1H),1.98(qt,J=13.8,3.6Hz,1H),1.57(s,3H),1.52–1.46(m,5H),1.40 (d,J=13.1Hz,1H),1.16(t,J=7.2Hz,6H),1.08(td,J=13.2,4.2Hz,1H),0.90(s,3H),0.72(s,3H).

[0229]

[0230] Compound II-4-5: colorless oil, 1 H NMR(600MHz, CDCl3) δ8.56–8.52(d,J=6.0Hz,2H),7.37(d,J=6.0Hz,2H),7.13(s,1H),3.95(dd,J=10 .2,6.6Hz,1H),3.88–3.78(m,2H),3.63(s,3H),3.39(d,J=13.2Hz,1H),2.96(hept,J=7.2Hz,1H),2. 55–2.41(m,1H),2.16–2.10(m,1H),2.05(tdd,J=17.4,7.8,4.8Hz,1H),1.64(s,4H),1.58(s,4H),1. 50–1.45(m,1H),1.23(dd,J=6.6,1.2Hz,7H),1.15(dd,J=12.0,2.4Hz,1H),0.97(s,3H),0.79(s,3H).

[0231]

[0232] Compound II-4-6: colorless oil, 1H NMR (600MHz, CDCl3) δ7.23–7.20(m,1H),7.17(s,1H),6.96–6.92(m,2H),4.03(d,J=6.6Hz,1H),4.01–3 .91(m,2H),3.61(s,3H),3.38(d,J=13.2Hz,1H),2.96(hept,J=7.2Hz,1H),2.50–2.42(m,1H),2.13(ddd ,J=12.6,6.6,1.8Hz,1H),2.05(dddd,J=17.4,13.8,9.0,3.6Hz,1H),1.64(s,3H),1.60–1.53(m,5H),1 .50–1.44(m,1H),1.25(dd,J=10.8,7.2Hz,6H),1.15(td,J=13.2,4.2Hz,1H),0.97(s,3H),0.79(s,3H).

[0233]

[0234] Compound II-4-7: white solid, 1 H NMR (600MHz, CDCl3) δ7.52(s,1H),6.88(s,1H),6.79(s,1H),4.09–4.04(m,1H),3.62(s,3H),3.40(d,J=13.8 Hz,1H),3.00(dddd,J=32.4,11.4,7.2,4.8Hz,2H),2.93(p,J=7.2Hz,1H),2.90–2.76(m,2H),2.45–2.36(m,1 H),2.20(ddd,J=12.6,7.2,2.4Hz,1H),2.02(qt,J=13.8,3.6Hz,1H),1.63(s,3H),1.58(m,5H),1.47(d,J=13 .2Hz,1H),1.31–1.24(m,2H),1.20(d,J=7.2Hz,6H),1.15(td,J=13.2,4.2Hz,1H),0.96(s,3H),0.76(s,3H).

[0235]

[0236] Compound II-4-8: white solid, 1H NMR (600MHz, CDCl3) δ6.92(s,1H),4.15(s,1H),3.73(s,4H),3.64(s,3H),3.40(d,J=13.8Hz,1H),2.92 (hept,J=7.2Hz,1H),2.75(ddt,J=24.0,12.0,5.4Hz,2H),2.63(dt,J=12.0,6.0Hz,1H),2.56(dt,J=12 .0,5.4Hz,1H),2.53–2.37(m,5H),2.15(s,1H),2.02(qt,J=13.8,3.6Hz,1H),1.63(s,3H),1.59(m,4H) ,1.47(d,J=13.2Hz,1H),1.23(t,J=7.2Hz,6H),1.15(td,J=13.2,4.2Hz,1H),0.97(s,3H),0.77(s,3H).

[0237]

[0238] Compound II-4-9: yield 78.2%, yellow solid, 1 H NMR(600MHz, CDCl3) δ6.93(s,1H),3.78(dd,J=10.2,6.6Hz,1H),3.53(s,3H),3.30(d,J=13 .2Hz,1H),3.12(d,J=11.4Hz,2H),2.61(t,J=12.0Hz,2H),2.47–2.40(m,2H),2.30(q,J=12 .0Hz,1H),2.03–1.93(m,2H),1.89(s,1H),1.79(d,J=12.6Hz,2H),1.56(s,3H),1.50(s,3H ),1.39(d,J=12.6Hz,1H),1.17–1.11(m,6H),1.10–1.03(m,1H),0.89(s,3H),0.70(s,3H).

[0239] Step 5: Synthesis of Compound II-5

[0240]

[0241] Synthesis of compounds II-5-1 to II-5-9: Under nitrogen, add the corresponding compounds II-4-1 to II-4-9 (approximately 200 mg, 0.50 mmol), sodium hydroxide (150 mg, 3.75 mmol), and anhydrous ethanol (10 mL) to a 25 mL round-bottom flask. Heat to 85°C and reflux for approximately 10 h. Completion of the reaction is monitored by TLC. Cool to room temperature, slowly add acetic acid (1 mL) to neutralize the sodium hydroxide, evaporate the solvent under reduced pressure, add saturated sodium bicarbonate solution (20 mL), extract with ethyl acetate (20 mL x 3), dry the extract over anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure. The residue is purified by silica gel column chromatography to obtain the corresponding product.

[0242]

[0243] Compound II-5-1, colorless oil, ESI-MS m / z: 370 [M+H] + , 1 H NMR (600MHz, CDCl3) δ7.00 (s, 1H), 6.36 (s, 1H), 4.34 (ddd, J = 5.4, 3.6, 2.4Hz, 1H) ,2.87(hept,J=7.2Hz,1H),2.66(d,J=12.6Hz,1H),2.02–1.88(m,3H),1.81–1.69( m,2H),1.61(s,3H),1.55(dd,J=9.6,3.0Hz,1H),1.50(s,3H),1.44(d,J=13.2Hz, 1H),1.18(t,J=4.8Hz,1H),1.11(dd,J=9.6,7.2Hz,6H),0.82(s,3H),0.75(s,3H). 13 C NMR (150MHz, CDCl3) δ177.07,143.09,141.44,135.06,125.61,119.89,115.95,110.42,51.71,47 .49,46.12,40.29,33.59,30.67,30.59,27.08,27.04,25.00,24.83,21.27,21.04,18.21,18.08.

[0244]

[0245] Compound II-5-2: colorless oil, 1HNMR(600MHz, CDCl3)δ7.19(d,J=7.8Hz,4H),7.12(d,J=7.8Hz,2H),6.18(s,1H),4.59(d,J=15.0Hz,1H), 4.29(d,J=15.0Hz,1H),4.17(t,J=2.4Hz,1H),2.91–2.80(m,1H),2.73(d,J=10.8Hz,1H),2.01(q,J=12.0 ,10.2Hz,2H),1.70(ddd,J=12.6,6.0,3.6Hz,1H),1.68–1.62(m,1H),1.60(s,3H),1.57(dd,J=10.2,5.4H z,2H),1.54(s,3H),1.43(d,J=13.2Hz,1H),1.18(s,1H),1.08(t,J=6.6Hz,6H),0.75(s,3H),0.70(s,3H).

[0246]

[0247] Compound II-4-3: white solid, 1 HNMR (600MHz, CDCl3) δ7.22(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.23(s,1H),4.55(d,J=15.0Hz,1H ),4.40(d,J=15.0Hz,1H),4.21(t,J=2.4Hz,1H),2.92(hept,J=7.2Hz,1H),2.79(d,J=10.8Hz,1H),2.1 2–2.00(m,2H),1.81–1.76(m,1H),1.76–1.70(m,1H),1.66(s,3H),1.64(dd,J=10.2,6.0Hz,2H),1.61( s,3H),1.50(d,J=13.2Hz,1H),1.25(t,J=13.2Hz,1H),1.15(t,J=6.6Hz,6H),0.80(s,3H),0.77(s,3H).

[0248]

[0249] Compound II-5-4: colorless oil, 1HNMR(600MHz, CDCl3)δ7.51(d,J=7.8Hz,2H),7.26(d,J=7.8Hz,2H),6.23(s,1H),4.63–4 .51(m,2H),4.25–4.17(m,1H),2.92(hept,J=7.2Hz,1H),2.80(d,J=11.2Hz,1H),2.15–2 .00(m,2H),1.84(m,1H),1.80–1.73(m,1H),1.67(s,3H),1.62(s,3H),1.51(d,J=13.2Hz ,1H),1.26(dd,J=8.4,4.2Hz,1H),1.14(dd,J=11.8,7.2Hz,6H),0.81(d,J=14.4Hz,6H).

[0250]

[0251] Compound II-5-5: colorless oil, 1 HNMR(600MHz, CDCl3)δ8.48(d,J=6.0Hz,2H),7.05(d,J=6.0Hz,2H),6.28(s,1H),4.58– 4.49(m,2H),4.22–4.19(m,1H),2.93(hept,J=7.2Hz,1H),2.80(d,J=11.4Hz,1H),2.13– 2.01(m,2H),1.91–1.85(m,1H),1.82–1.76(m,1H),1.68(s,3H),1.63(s,3H),1.52(d,J =13.8Hz,1H),1.28–1.23(m,2H),1.16(dd,J=6.6,4.2Hz,6H),0.83(s,3H),0.81(s,3H).

[0252]

[0253] Compound II-5-6: colorless oil, 1HNMR (600MHz,CDCl3) δ7.21(dd,J=4.8,1.2Hz,1H),6.95(d,J=2.4Hz,1H),6.93(dd,J=4.8,3.6Hz,1H),6.34(s,1H),4.97(d,J=15.0Hz,1H),4.39–4.34(m,2H),2.93(hept,J=7.2Hz,1H),2. 78(d,J=7.8Hz,1H),2.06(m,2H),1.89–1.82(m,1H),1.78–1.70(m,1H),1.67(s,3H),1.64(m,2H),1.59( s,3H),1.49(d,J=11.2Hz,1H),1.25(t,J=13.2Hz,1H),1.17(t,J=7.2Hz,6H),0.81(s,3H),0.77(s,3H).

[0254]

[0255] Compound II-5-7: white solid, 1 HNMR(600MHz, CDCl3)δ7.63(s,1H),6.68(s,1H),6.38(s,1H),4.38–4.31(m,1H),3.66(m,1H),3.50(m,1H),3 .05(q,J=7.2Hz,4H),2.86(m,2H),2.77(m,1H),2.63(d,J=13.8Hz,1H),2.01(td,J=13.8,4.2Hz,1H),1.92(q ,J=13.2Hz,1H),1.86–1.80(m,1H),1.73(t,J=12.6Hz,1H),1.59(s,3H),1.56(dd,J=10.2,6.0Hz,2H),1.52( s,3H),1.40(d,J=13.2Hz,2H),1.32(t,J=7.2Hz,6H),1.12(dd,J=6.6,4.8Hz,6H),0.71(s,3H),0.64(s,3H).

[0256]

[0257] Compound II-5-8: yellow oil, 1H NMR (600MHz, CDCl3) δ6.44(s,1H),4.38–4.33(m,1H),3.69–3.62(m,4H),3.59(dt,J=1 3.8,6.0Hz,1H),3.34(m,1H),2.95(hept,J=7.2Hz,1H),2.74(d,J=11.4Hz,1H),2.52–2 .38(m,5H),2.08–2.00(m,3H),1.84–1.78(m,1H),1.67(s,3H),1.66–1.60(m,2H),1.5 8(s,3H),1.49(d,J=13.2Hz,1H),1.19(dd,J=11.4,6.6Hz,6H),0.82(d,J=13.2Hz,6H).

[0258]

[0259] Compound II-5-9: 77.3%, yield: colorless oil, 1 H NMR(600MHz, CDCl3)δ6.44(s,1H),4.28–4.23(m,1H),3.33(dd,J=13.8,7.8Hz,1H),3.08–2.98(m,3H),2.9 5(p,J=7.2Hz,1H),2.74(d,J=10.2Hz,1H),2.54(td,J=12.0,2.4Hz,1H),2.44(td,J=12.0,2.4Hz,1H),2.09 –2.00(m,2H),1.96(ddd,J=12.6,5.4,3.6Hz,1H),1.85–1.79(m,1H),1.66(s,3H),1.59(s,3H),1.49(d,J=1 2.0Hz,1H),1.40(d,J=13.8Hz,1H),1.18(dd,J=14.4,6.6Hz,7H),1.13–1.05(m,2H),0.81(d,J=2.4Hz,6H).

[0260] Step 6: Synthesis of Compound II-6

[0261]

[0262] Synthesis of compounds II-6-1 to II-6-8: Add the corresponding compounds II-5-1 to II-5-8 (approximately 100 mg, 0.27 mmol) to dichloromethane (5 mL), stir to dissolve, then slowly add trifluoroacetic acid (5 mL), stir at room temperature overnight, and monitor the reaction completion by TLC. Evaporate the solvent under reduced pressure, add saturated sodium bicarbonate solution (20 mL) to the residue, extract with dichloromethane (20 mL × 3), dry the extract with anhydrous sodium sulfate, filter, and evaporate the dichloromethane under reduced pressure. The residue is purified by silica gel column chromatography to obtain the corresponding products II-6-1 to II-6-8.

[0263]

[0264] Compound II-6-1: white solid, ESI-MS m / z: 328 [MH] - , 1 H NMR(600MHz,DMSO-d6)δ8.17(d,J=5.4Hz,1H),8.09(s,1H),7.82(s,1H),6.52(s,1H),4.37 –4.23(m,1H),3.25–3.14(m,1H),2.68(d,J=13.2Hz,1H),2.32(td,J=13.8,4.2Hz,1H),1.9 2–1.77(m,2H),1.63(t,J=10.8Hz,1H),1.50(dd,J=10.2,5.4Hz,1H),1.45(d,J=13.2Hz,1H ),1.40(d,J=12.6Hz,1H),1.21–1.15(m,1H),1.10(t,J=6.6Hz,6H),0.79(d,J=16.2Hz,6H). 13 C NMR(150MHz,DMSO-d6)δ177.04,143.67,141.65,136.82,133.43,125.07,110.07,51.7 4,50.01,47.24,41.60,34.98,32.11,31.75,29.24,26.58,23.42,23.20,19.94,19.57.

[0265]

[0266] Compound II-6-2: white solid, ESI-MS m / z: 418 [MH] - , 11H NMR (600 MHz, CDCl3) δ 7.17–7.11 (m, 3H), 6.99 (d, J = 6.6 Hz, 2H), 6.20 (s, 1H), 4.46–4.37 (m, 2H), 4.18–4.12 (m, 1H), 3.09–2.94 (m, 2H), 2.31 (td, J = 12.6, 3.6 Hz, 1H), 2.02 (q, J = 13.8 Hz, 1H), 1.74 (dt, J = 10.8, 3.6 Hz, 1H), 1.66–1.58 (m, 4H), 1.44 (d, J = 13.2 Hz, 1H), 1.21–1.16 (m, 2H), 1.04 (d, J = 7.2 Hz, 3H), 1.00 (d, J = 7.2 Hz, 3H), 0.73 (s, 3H), 0.70 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.37, 140.79, 139.60, 135.53, 133.43, 131.19, 127.54, 126.31, 123.49, 110.35, 56.45, 49.56, 47.23, 46.52, 40.28, 33.88, 29.46, 28.35, 25.86, 21.46, 18.85, 18.33.

[0267]

[0268] Compound II-6-3: White solid, ESI-MS m / z: 453 [M-H] - , 1 1H NMR (600 MHz, CDCl3) δ 7.04 (d, J = 8.​C NMR (150MHz, CDCl3) δ175.68,141.95,140.83,135.11,134.41,133.20,132.50,124.49,111.3 2,57.80,50.65,47.69,47.53,41.29,34.94,31.63,30.52,29.31,26.93,22.51,19.88,19.35.

[0269]

[0270] Compound II-6-4: ESI-MS m / z: 486 [MH] - , 1 H NMR (600MHz, CDCl3) δ7.35 (d, J = 8.4Hz, 2H), 7.05 (d, JII-6-4 = 7.8Hz, 2H), 6.16 (s, 1H), 4.59 (d, J = 15. 0Hz,1H),4.35(d,J=15.0Hz,1H),4.13–4.09(m,1H),3.03(d,J=12.0Hz,1H),2.97(hept,J=7.2Hz,1H) ,2.32(td,J=13.2,4.2Hz,1H),2.00(qt,J=13.8,3.0Hz,1H),1.79(dt,J=12.0,4.1Hz,1H),1.71–1.5 9(m,3H),1.45(d,J=13.2Hz,1H),1.02(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H),0.73(d,J=6.0Hz,6H). 13 C NMR (150MHz, CDCl3) δ174.61,140.69,139.75,133.38,131.38,127.63,124.25,123.21,110.25,57.00,4 9.57,46.92,46.45,40.21,33.89,30.57,30.40,29.49,29.15,28.33,25.87,21.39,21.33,18.79,18.29.

[0271]

[0272] Compound II-6-5: ESI-MS m / z: 419 [MH] - , 1H NMR (600MHz, CDCl3) δ8.29(d,J=6.0Hz,2H),6.92(d,J=6.0Hz,2H),6.29(s,1H),4.81(d,J=15.6Hz,1H),4.27(d,J=1 5.6Hz,1H),4.22–4.17(m,1H),3.15(hept,J=6.6Hz,1H),3.00(d,J=13.8Hz,1H),2.43(td,J=13.2,4.2Hz,1H),2.01 (qt,J=13.8,3.0Hz,1H),1.93(ddd,J=12.6,5.4,3.6Hz,1H),1.83–1.77(m,1H),1.72(dd,J=10.2,5.4Hz,1H),1.64( dt,J=13.8,3.6Hz,1H),1.50(d,J=13.2Hz,1H),1.10(d,J=6.6Hz,3H),1.02(d,J=6.6Hz,3H),0.81(d,J=3.0Hz,6H). 13 C NMR (150MHz, CDCl3) δ175.84,148.89,147.20,142.05,141.77,134.64,133.14,124.15,123.29,110.8 7,58.81,50.52,47.72,47.44,41.26,34.92,31.67,30.64,29.20,26.90,22.72,22.68,19.75,19.33.

[0273]

[0274] Compound II-6-6: yellow solid, ESI-MS m / z: 424 [MH] - , 11H NMR (600 MHz, CDCl3) δ 7.09 (dd, J = 4, 8, 1.2 Hz, 1H), 6.81 (dd, J = 4.8, 3.6 Hz, 1H), 6.77 (d, J = 3.0 Hz, 1H), 6.31 (s, 1H), 4.72 (d, J = 15.0 Hz, 1H), 4.35 (d, J = 15.0 Hz, 1H), 4.29–4.22 (m, 1H), 3.09–2.98 (m, 2H), 2.30 (td, J = 13.2, 3.6 Hz, 1H), 1.98 (q, J = 13.8 Hz, 1H), 1.79 (ddd, J = 12.6, 5.4, 3.6 Hz, 1H), 1.69–1.63 (m, 1H), 1.62–1.56 (m, 2H), 1.42 (d, J = 12.6 Hz, 1H), 1.07 (d, J = 7.2 Hz, 3H), 1.03 (d, J = 6.6 Hz, 3H), 0.70 (d, J = 6.0 Hz, 6H). 13 13C NMR (150 MHz, CDCl3) δ 174.14, 140.91, 139.76, 137.94, 133.38, 131.23, 126.07, 125.61, 124.34, 123.38, 110.39, 56.51, 49.67, 46.42, 41.73, 40.29, 33.86, 30.58, 29.77, 28.27, 25.93, 21.55, 21.47, 18.81, 18.32.

[0275]

[0276] Compound II-6-7, ESI-MS m / z: 422 [M-H] - , 1 1H NMR (600 MHz, Methanol-d4) δ 7.54 (s, 1H), 6.49 (s, 1H), 6.45 (s, 1H), 4.39–4.34 (m, 1H), 3.55–3.45 (m, 2H), 3.12 (hept, J = 6.6 Hz, 1H), 2.73 (d, J = 14.4 Hz, 1H), 2.65 (t, J = 7.2 Hz, 2H), 2.40 (td, J = 13.8, 4.2 Hz, 1H), 1.91–1.80 (m, 2H), 1.61 (t, J = 11.4 Hz, 1H), 1.52–1.43 (m, 2H), 1.35 (d, J = 14.4 Hz, 1H), 1.08 (dd, J = 7.2, 4.8 Hz, 6H), 0.69 (s, 3H), 0.66 (s, 3H). 13C NMR(150MHz,Methanol-d4)δ176.37,143.03,141.51,135.22,134.45,134.07,133.53,124.10,116.53,109.8 3,58.75,50.31,48.47,44.58,41.14,34.49,30.85,30.40,28.89,26.57,25.15,22.01,21.90,19.18,18.72.

[0277]

[0278] Compound II-6-8, white solid, ESI-MS m / z: 441 [MH] - , 1 H NMR (600MHz, CDCl3) δ6.52 (s, 1H), 4.36–4.29 (m, 1H), 3.70–3.58 (m, 4H), 3.50 (dt, J = 13.2, 6.0Hz,1H),3.32(dt,J=13.8,7.2Hz,1H),3.18(hept,J=6.6Hz,1H),3.03(d,J=13.2Hz,1H), 2.47–2.35(m,7H),2.07–1.98(m,2H),1.82–1.76(m,1H),1.69–1,62(m,2H),1.49(d,J=13. 2Hz, 1H), 1.26 (td, J = 13.2, 4.2Hz, 1H), 1.17 (dd, J = 15.6, 7.2Hz, 6H), 0.81 (d, J = 4.2Hz, 6H). 13 C NMR (150MHz, CDCl3) δ175.40,142.15,141.13,134.68,132.46,124.82,111.08,66.85,59.47,56.51 ,53.55,50.43,47.36,42.44,41.29,34.95,31.74,30.88,29.19,27.07,22.64,22.58,19.85,19.36.

[0279] Example 4: Synthesis of carnosol derivatives based on compound II-6-1

[0280]

[0281] Synthesis of Compound II-7-1: Under nitrogen, compound II-6-1 (100 mg, 0.30 mmol), potassium carbonate (126 mg, 0.90 mmol), and acetone (5 mL) were added to a 25 mL single-necked flask and stirred until uniformly dispersed. Methyl iodide (127 mg, 0.90 mmol) was then added dropwise. The mixture was heated to 65°C and refluxed for 20 h. Completion of the reaction was monitored by TLC. The mixture was cooled to room temperature, filtered under reduced pressure to remove the potassium carbonate, and the filtrate was evaporated to dryness under reduced pressure. Saturated ammonium chloride solution (20 mL) was added to the residue, and the mixture was extracted with ethyl acetate (20 mL x 3). The extract was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, volume ratio 7:2) to obtain 84 mg of a yellow solid (II-7-1) in a 77.0% yield. ESI-MS m / z: 358 [M+H] + , 1 H NMR (600MHz, CDCl3) δ6.65(s,1H),4.36(s,1H),3.71(s,3H),3.70(s,3H),3.19(hep t,J=6.6Hz,1H),2.77(d,J=12.6Hz,1H),2.29(td,J=13.8,4.2Hz,1H),1.94(dd,J=13 .2,3.6Hz,2H),1.78–1.70(m,1H),1.62–1.54(m,2H),1.44(d,J=13.2Hz,1H),1.21– 1.15(m,4H),1.11(d,J=7.2Hz,3H),1.07(d,J=7.2Hz,3H),0.82(s,3H),0.76(s,3H). 13 C NMR (150MHz, CDCl3) δ177.39,149.73,149.18,139.70,138.41,130.17,113.55,60.05,59.64 ,51.68,49.53,45.81,40.30,33.97,30.79,30.15,27.06,25.56,22.67,22.38,18.61,18.45.

[0282] Synthesis of Compound II-7-2: Under nitrogen, compound II-6-1 (550 mg, 1.67 mmol) and 1,4-dioxane (10 mL) were added to a 25 mL single-necked flask and stirred to dissolve. DDQ (400 mg, 1.76 mmol) was then slowly added and stirred at room temperature for 30 min. The reaction was monitored by TLC until complete. The reaction mixture was filtered under reduced pressure, and the filter cake was washed with dichloromethane (10 mL). The filtrate was evaporated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane-methanol, volume ratio 100:1) to obtain 515 mg of a dark green solid (II-7-2), with a yield of 93.6%. ESI-MS m / z: 329 [M+H] + , 1 H NMR(600MHz, CDCl3)δ8.19(s,1H),6.59(s,1H),4.23(s,1H),2.84(hept,J=6 .6Hz,1H),2.60(d,J=14.4Hz,1H),2.18(td,J=13.8,4.2Hz,1H),1.96–1.90( m,1H),1.90–1.77(m,2H),1.60–1.50(m,2H),1.42(d,J=13.2Hz,1H),1.14(t d,J=13.8,3.6Hz,1H),1.01(dd,J=10.8,7.2Hz,6H),0.78(d,J=12.6Hz,6H). 13 C NMR (150MHz, CDCl3) δ179.94,177.30,176.55,156.22,149.97,137.40,130.92,52.39 ,50.82,46.52,40.84,34.82,32.14,29.54,27.66,26.90,21.49,21.35,19.11,18.83.

[0283] Synthesis of compounds II-8-1 to II-8-4: Under nitrogen, compound II-7-2 (150 mg, 0.46 mmol), ammonium acetate (350 mg, 4.6 mmol), paraformaldehyde (70 mg, equivalent to 2.3 mmol of formaldehyde), and acetic acid (10 mL) were added to a 25 mL single-necked flask. The mixture was stirred and dissolved, heated to 110°C, and refluxed for 10 h. The reaction was monitored by TLC for completion. The mixture was cooled to room temperature and the acetic acid was evaporated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with ethyl acetate (20 mL x 3). The extract was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane-methanol, volume ratio 150:1 to 80:1) to obtain 40 mg of compound a and 76 mg of compound b, with a total yield of 65.4%. Mixture a was purified by HPLC (methanol-water, volume ratio 8:2) to obtain 25 mg of compound II-8-1 and 9 mg of compound II-8-2, and mixture b was purified by HPLC (methanol-water, volume ratio 8:2) to obtain 50 mg of compound II-8-3 and 13 mg of compound II-8-4.

[0284] Compound II-8-1: white solid, ESI-MS m / z: 361 [M+Na] + , 1 H NMR (600MHz, CDCl3) δ8.02(s,1H),7.34(d,J=4.8Hz,1H),7.03(s,1H),4.68(ddd,J=5.4,3.6,2.4H z,1H),3.57(hept,J=6.6Hz,1H),3.01(d,J=13.8Hz,1H),2.37(td,J=13.2,4.2Hz,1H),2.18–2.06 (m,2H),1.92–1.85(m,1H),1.75(dt,J=13.8,3.6Hz,1H),1.65(dd,J=10.2,5.4Hz,1H),1.58(d,J= 13.2Hz, 1H), 1.33 (dd, J=13.2, 6.6Hz, 6H), 1.28 (dd, J=13.8, 3.6Hz, 1H), 0.96 (s, 3H), 0.85 (s, 3H). 13 C NMR (150MHz, CDCl3) δ177.65,151.36,145.31,140.68,139.38,138.02,123.81,115.11,52 .91,48.93,47.01,41.27,34.67,31.75,31.52,29.34,28.23,23.15,22.97,19.22,19.15.

[0285] Compound II-8-2: white solid, ESI-MS m / z: 361 [M+Na] + , 1 H NMR (600MHz, CDCl3) δ8.07(s,1H),7.03(s,1H),6.65(d,J=4.8Hz,1H),4.65(ddd,J=5.4,3.6,2. 4Hz,1H),3.34(hept,J=6.6Hz,1H),3.10(d,J=13.8Hz,1H),2.69(td,J=13.8,4.2Hz,1H),2.16–2 .07(m,2H),1.87(t,J=11.4Hz,1H),1.80(dp,J=13.8,3.6Hz,1H),1.66(dd,J=10.2,5.4Hz,1H),1 .56(d,J=13.2Hz,1H),1.34(dd,J=6.6,6.0Hz,6H),1.31–1.24(m,1H),0.96(s,3H),0.85(s,3H). 13 C NMR (150MHz, CDCl3) δ177.70,151.93,147.73,139.42,136.18,132.02,129.89,116.39 ,52.89,50.40,46.89,41.29,34.73,31.82,29.18,28.81,22.55,22.42,19.37,19.16.

[0286] Compound II-8-3: white solid, ESI-MS m / z: 432 [M+Na] + , 1 H NMR (600MHz, CDCl3) δ8.02(s,1H),7.09(s,1H),6.88(q,J=7.8,6.6Hz,1H),5.08(dd,J=3.6,2.4Hz,1 H),4.72(qd,J=13.8,6.6Hz,2H),3.57(hept,J=6.6Hz,1H),2.96(d,J=13.8Hz,1H),2.37(td,J=13.2 ,4.2Hz,1H),2.15–2.02(m,2H),1.89(s,3H),1.84(ddd,J=12.6,10.4,2.4Hz,1H),1.72(dt,J=13.8, 3.6Hz, 1H), 1.62 (dd, J = 10.2, 5.4Hz, 1H), 1.55 (d, J = 13.2Hz, 1H), 1.35 (t, J = 7.2Hz, 6H), 0.83 (s, 6H). 13C NMR (150MHz, CDCl3) δ175.52,171.34,151.34,145.12,140.37,139.59,138.15,123.07,115.21,59.5 0,49.78,49.01,46.92,41.18,34.71,31.63,30.77,29.38,28.33,23.22,22.97,22.86,19.17,19.07.

[0287] Compound II-8-4: white solid, ESI-MS m / z: 432 [M+Na] + , 1 H NMR (600MHz, CDCl3) δ8.07 (s, 1H), 7.08 (s, 1H), 6.66 (s, 1H), 5.04 (dd, J = 3.6, 2.4Hz, 1H) ,4.75(qd,J=13.8,6.6Hz,2H),3.34(hept,J=6.6Hz,1H),3.05(d,J=15.0Hz,1H),2.70(td ,J=13.8,4.2Hz,1H),2.14–2.00(m,2H),1.88(s,3H),1.85–1.76(m,3H),1.61(dd,J=10. 2,5.4Hz,1H),1.54(d,J=13.2Hz,1H),1.34(dd,J=7.2,3.6Hz,6H),0.83(d,J=7.8Hz,6H). 13 CNMR (150MHz, CDCl3) δ175.91,171.22,151.91,147.87,139.18,135.95,131.23,130.06,116.54,59.3 8,50.42,49.84,46.85,41.20,34.79,31.71,30.89,29.21,28.89,22.94,22.60,22.44,19.28,19.11.

[0288] Example 5: Synthesis of 6-Modified Carnosol Derivatives

[0289]

[0290] Synthesis of Compound III-1: Carnosol I-2 (5.0 g, 15.10 mmol) was added to a 150 mL round-bottom flask. Under nitrogen, 50 mL of acetone was added and stirred to dissolve. Potassium carbonate (6.3 g, 45.65 mmol) was slowly added, followed by dropwise addition of iodomethane (12.9 g, 90.90 mmol). The mixture was heated to 65°C and refluxed for 48 h. After TLC analysis, the solid was removed by filtration. The filter cake was washed with 20 mL of dichloromethane and the solvent was evaporated to dryness. The residue was added with 50 mL of ethyl acetate and 50 mL of saturated ammonium chloride solution, shaken, and separated. The aqueous phase was extracted twice with 50 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 80:1) to afford III-1 (5.08 g, 90.40% yield) as a colorless oil. 1 H NMR(600MHz,Chloroform-d)δ6.67(s,1H),6.36(dd,J=9.6,3.0Hz,1H),6.03(dd,J=9.6,3.0Hz,1H),3.78 (s,3H),3.75(s,3H),3.71(d,J=13.2Hz,1H),3.53(s,3H),3.24(p,J=7.2Hz,1H),2.55(t,J=3.0Hz,1H),1. 81(qt,J=13.8,3.6Hz,1H),1.64(dp,J=14.4,3.7Hz,1H),1.55(td,J=13.2,3.6Hz,1H),1.47(d,J=13.2Hz, 1H),1.29(td,J=13.2,4.2Hz,1H),1.21(d,J=7.2Hz,3H),1.18(d,J=7.2Hz,3H),1.01(s,3H),0.88(s,3H). 13 C NMR(150MHz,Chloroform-d)δ174.27,152.86,150.56,141.71,131.91,130.22,127.90,127.08,120.4 9,60.41,60.01,51.60,51.25,49.53,40.97,34.30,33.71,32.03,26.63,23.61,23.06,21.60,20.57.

[0291] Synthesis of Compound III-2: Compound III-1 (3.0 g, 8.06 mmol) was added to a 150 mL round-bottom flask. Under nitrogen, 30 mL of acetonitrile was added and stirred to dissolve. 30 mL of water was then added, the temperature was lowered to 0°C, and stirring was continued for 10 min. m-Chloroperbenzoic acid (2.1 g, 12.10 mmol) was then added, and stirring was continued at 0°C for 2 h. After TLC analysis, the acetonitrile was evaporated under reduced pressure, 30 mL of saturated sodium bicarbonate solution was added, and the aqueous phase was extracted three times with 50 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain 2.71 g of a white solid, with a yield of 82.7%. 1 H NMR (600MHz, Chloroform-d) δ7.18 (s, 1H), 4.50 (dd, J = 11.4, 8.4Hz, 1H), 4.44 (d ,J=8.4Hz,1H),3.74(s,3H),3.66(d,J=5.4Hz,6H),3.36(dt,J=14.4,7.8Hz,1H) ,3.25(p,J=7.2Hz,1H),1.90–1.82(m,1H),1.78(d,J=11.4Hz,2H),1.57–1.50(m ,1H),1.36–1.31(m,1H),1.25(s,3H),1.22(dd,J=11.8,7.2Hz,6H),0.97(s,3H). 13 C NMR(150MHz,Chloroform-d)δ176.84,150.62,149.97,142.09,133.79,132.69,118.81,72.56,60 .07,59.18,51.90,51.47,50.58,39.26,34.13,33.49,33.12,26.95,23.55,23.29,22.31,18.47.

[0292] Synthesis of Compound III-3: Compound III-2 (2.8 g, 6.90 mmol) was added to a 50 mL round-bottom flask. Under nitrogen, 30 mL of methanol was added and stirred at room temperature to dissolve. Sodium hydroxide (600 mg, 15 mmol) was added and stirred at room temperature for 1 h. After TLC analysis, the reaction was complete. 1 mL of acetic acid was added to neutralize the sodium hydroxide. The solvent was evaporated under reduced pressure. 30 mL of ethyl acetate and 30 mL of saturated sodium bicarbonate solution were added to the residue. After shaking, the liquids were separated. The aqueous phase was extracted twice with 30 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 7:1) to obtain 1.71 g of a white solid with a yield of 66.30%. 1H NMR (600MHz, Chloroform-d) δ6.98 (s, 1H), 5.22 (d, J = 4.5Hz, 1H), 4.48–4.26 (m, 1H), 3. 82(s,3H),3.79(s,3H),3.30(h,J=7.2Hz,1H),2.82–2.73(m,1H),2.41(td,J=13.8,4.2 Hz,1H),1.91(qt,J=13.8,3.0Hz,1H),1.75–1.60(m,2H),1.56–1.48(m,1H),1.31(d,J= 3.6Hz, 1H), 1.28 (dd, J=13.8, 3.0Hz, 1H), 1.24–1.17 (m, 6H), 1.05 (s, 3H), 0.94 (s, 3H). 13 C NMR(150MHz,Chloroform-d)δ174.59,152.45,150.72,142.50,130.91,128.83,119.80,79.29,68 .45,61.03,60.60,56.87,48.30,40.85,34.45,32.17,28.12,26.85,23.52,23.28,21.00,18.81.

[0293]

[0294] Synthesis of compound III-4-1: Compound III-3 (200 mg, 0.53 mmol) was added to a 25 mL round-bottom flask. After nitrogen replacement of the air, 10 mL of dry dichloromethane and pyridine (125 mg, 1.60 mmol) were added. The temperature of the system was lowered to 0 ° C. After stirring for 10 min, benzoyl chloride (150 mg, 1.06 mmol) was slowly added dropwise. After the dropwise addition, the temperature of the system was slowly raised to room temperature and stirred at room temperature for 24 h. After TLC detection, 10 mL of dilute hydrochloric acid (1 mol / L) was added and stirred for 30 min to quench the reaction. 20 mL of dichloromethane was added to dilute and 20 mL of dilute hydrochloric acid (1 mol / L) was added. The organic phase was separated and washed with 30 mL of saturated sodium bicarbonate. After separation, anhydrous sodium sulfate was added to dry the organic phase. After filtration, the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 30: 1) to give 150 mg of a white solid with a yield of 58.60%. 1H NMR(600MHz,Chloroform-d)δ7.73(dd,J=8.4,1.2Hz,2H),7.53(t,J=7.2Hz,1H),7.35(t,J=7.8Hz,2H),6.82 (s,1H),5.86–5.66(m,1H),5.50(d,J=4.8Hz,1H),3.87(s,3H),3.84(s,3H),3.37–3.24(m,1H),2.86(d,J=15. 6Hz,1H),2.49(td,J=13.8,4.2Hz,1H),1.97(qt,J=13.8,3.0Hz,1H),1.70(dd,J=9.6,3.6Hz,2H),1.55(d,J=1 2.0Hz,1H),1.32(td,J=13.8,3.6Hz,1H),1.26(s,2H),1.13(d,J=6.6Hz,3H),1.08–1.01(m,6H),0.95(s,3H). 13 C NMR(150MHz,Chloroform-d)δ174.20,165.54,152.25,150.70,142.66,133.36,131.56,129.52,129.50,128.34,128.24, 118.84,75.98,70.25,61.17,60.78,52.80,48.28,40.67,34.48,31.76,29.71,28.15,26.60,23.52,23.27,20.87,18.79.

[0295]

[0296] Synthesis of compound III-4-2: Nicotinic acid (72 mg, 0.58 mmol), N,N-diisopropylethylamine (165 mg, 1.32 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (302.30 mg, 0.79 mmol) were added to a 25 mL reaction bottle. Under nitrogen protection, 10 mL of dry DMF was added. After stirring at room temperature for 0.5 h, compound III-3 (200 mg, 0.53 mmol) was added and stirring was continued for 8 h. After TLC detection of compound 43 disappeared, 50 mL of water and 20 mL of saturated brine were added, and the mixture was cooled to 0°C to precipitate a yellow solid. The solid was collected by filtration, and 30 mL of dichloromethane was added to the solid residue to dissolve it. The mixture was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to obtain 94 mg of a white solid in a yield of 36.72%. 1H NMR(600MHz,Chloroform-d)δ8.85(d,J=2.4Hz,1H),8.74(dd,J=4.8,1.7Hz,1H),8.05(dt,J=7.8,1.8Hz,1H), 7.33(dd,J=8.4,5.4Hz,1H),6.81(s,1H),5.80–5.68(m,1H),5.52(d,J=4.8Hz,1H),3.86(s,3H),3.84(s,3H), 3.29(p,J=7.2Hz,1H),2.86(d,J=15.6Hz,1H),2.50(td,J=13.8,4.2Hz,1H),1.97(qt,J=13.8,3.0Hz,1H),1.7 1(dd,J=12.0,3.6Hz,2H),1.34(td,J=13.8,3.6Hz,1H),1.13(d,J=6.6Hz,3H),1.05–1.00(m,6H),0.95(s,3H).

[0297] Example 6: Experiment on the effect of carnosol derivatives on the atrophy of mouse myoblasts (C2C12) induced by the supernatant of mouse colon cancer cell (C26) culture fluid

[0298] The diameter of differentiated mouse myoblasts (C2C12) was measured using a diameter measurement method. The staining method used was hematoxylin-eosin (HE) staining. Hematoxylin, an alkaline, positively charged dye, readily binds ionically to the negatively charged, acidic deoxyribonucleic acid (DNA) in the cell nucleus, staining it blue. Eosin, an acidic dye, dissociates in water into negatively charged anions and readily binds to the positively charged amino groups of proteins in the cytoplasm, staining them red. After staining, the cells were imaged under a high-magnification microscope, and the diameters of the myotubes were calculated using Digimizer software.

[0299] The specific method is as follows: C2C12 cells were seeded at 30,000 cells / mL in a 24-well plate and differentiated into mature myotubes using high-glucose DMEM medium supplemented with 2% HS and 1% PS. Separately, C26 cells and 3T3-L1 adipocytes were seeded in T75 flasks and cultured in high-glucose DMEM supplemented with 10% FBS and 1% PS in a 5% CO2, 37°C cell culture incubator. Six million cells were passaged per flask. After 48 hours of incubation in 20 mL of culture medium, the supernatant was collected and centrifuged at 1000 rpm for 3 minutes. The supernatant was then collected and centrifuged at 4000 rpm for 10 minutes. The 3T3-L1 supernatant was mixed with 2% HS differentiation medium in a 1:1 ratio to serve as the healthy control group. The C26 supernatant was mixed with 2% HS differentiation medium in a 1:1 ratio to serve as the muscle atrophy-inducing medium. Equal volumes of the muscle atrophy-inducing medium were added to both the model and treatment groups. At the same time, different stock solutions of carnosol and carnosol derivatives were added to the cells at the following concentrations: 12.5 μM or 25 μM, see Table 1.

[0300] Table 1 shows the dosage regimen of carnosol and carnosol derivatives in the experiment on reversal of muscle atrophy in C2C12 muscle cells in Example 6.

[0301] Table 1. Samples administered for the myoblast (C2C12) atrophy experiment in Example 6

[0302]

[0303]

[0304] Here, μM refers to μMol / L.

[0305] The myotube diameter was measured as follows:

[0306] After 48 hours of drug treatment, the cells were fixed with a fixative (anhydrous ethanol: formaldehyde: glacial acetic acid = 20:2:1) for more than 1 hour, stained with hematoxylin-eosin, and images were collected under a high-power microscope. Digimizer was used to calculate the myotube diameter.

[0307] The muscle atrophy reversal rate was calculated according to the following formula:

[0308] Muscle atrophy reversal rate = (average value of myotubes in the drug group - average value of myotubes in the model group) / (average value of myotubes in the control group - average value of myotubes in the model group) × 100%

[0309] Results and conclusions: Please refer to the attached Figure 1-54 Attached Figure 1-54 The following are representative HE staining images of carnosol and carnosol derivatives alleviating C2C12 mature myotube atrophy induced by C26 cell culture medium. Appendix 2 is the statistical results of myotubes. Figure 1-54As shown in Table 2, carnosol derivatives have a significant reversal effect on muscle cell atrophy.

[0310] Table 2 shows the statistical results of carnosol and carnosol derivatives in alleviating C2C12 muscle cell atrophy, corresponding to the attached Figure 1-54 .

[0311] Table 2. Reversal rate of muscle atrophy in C2C12 myotube cells treated with carnosol and carnosol derivatives

[0312]

[0313]

[0314] Example 7: Effect of I-4-6, II-6-2, II-8-1 and II-8-2 on the Viability of Mouse Myoblasts (C2C12)

[0315] The MTT assay was used to assess C2C12 cell viability. The assay works by reducing exogenous MTT to water-insoluble, blue-purple formazan crystals in viable cells, while dead cells do not. A triple solution (10% SDS, 5% isobutanol, and 0.01% concentrated hydrochloric acid in water) dissolves the formazan in the cells. The absorbance at 570 nm is measured using an enzyme-linked immunosorbent assay (ELISA), indirectly reflecting the number of viable cells. Within a certain cell population range, the amount of MTT crystals formed is proportional to the cell number.

[0316] The above method can be used to evaluate the effect of drugs on the viability of mouse myoblasts (C2C12). The specific method is as follows:

[0317] C2C12 cells were seeded at 30,000 cells / mL in a 24-well plate and cultured in high-glucose DMEM supplemented with 10% FBS and 1% PS in a 5% CO2, 37°C incubator. When the cell density reached 70%-80%, the culture medium was replaced with high-glucose DMEM supplemented with 2% HS and 1% PS. The differentiation medium was changed every 48 hours, and myotubes matured on day 5 or 6. I-4-6, II-6-2, II-8-1, or II-8-2 were added to the differentiation medium at the following concentrations: 1.0625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, or 100 μM. The differentiated C2C12 myotubes were treated for 48 hours. The culture medium in the plate was discarded, and 200 μL of fresh DMEM medium without FBS was added. 20 μL of MTT was added to each well and incubated in the cell culture incubator for 4 h. After 4 h, 50 μL of the triplicate solution was added to each well and incubated overnight. The absorbance was then measured at a wavelength of 570 nm using a microplate reader.

[0318] Cell viability (%) = (OD drug -OD blank ) / (OD control -OD blank )×100%

[0319] Results and conclusions: Please refer to the attached Figures 55-58 , attached Figures 55-58 The following table shows the survival rates of C2C12 myotube cells treated with different concentrations of I-4-6, II-6-2, II-8-1, and II-8-2 for 48 hours. As shown in Table 3, when the I-4-6 concentration was 1.0625 μM, the survival rate was 105.959%; at 3.125 μM, the survival rate was 102.148%; at 6.25 μM, the survival rate was 93.721%; at 12.5 μM, the survival rate was 107.047%; at 25 μM, the survival rate was 103.457%; at 50 μM, the survival rate was 102.811%; and at 100 μM, the survival rate was 98.752%. When the concentration of Ⅱ-6-2 was 1.0625 μM, the survival rate was 101.057%; when the concentration was 3.125 μM, the survival rate was 98.091%; when the concentration was 6.25 μM, the survival rate was 102.223%; when the concentration was 12.5 μM, the survival rate was 103.822%; when the concentration was 25 μM, the survival rate was 101.669%; when the concentration was 50 μM, the survival rate was 69.800%; when the concentration was 100 μM, the survival rate was 11.228%. When the concentration of Ⅱ-8-1 was 1.0625 μM, the survival rate was 101.778%; when the concentration was 3.125 μM, the survival rate was 108.838%; when the concentration was 6.25 μM, the survival rate was 109.828%; when the concentration was 12.5 μM, the survival rate was 102.572%; when the concentration was 25 μM, the survival rate was 104.249%; when the concentration was 50 μM, the survival rate was 98.067%; when the concentration was 100 μM, the survival rate was 7.763%. When the concentration of Ⅱ-8-2 was 1.0625 μM, the survival rate was 105.227%; when the concentration was 3.125 μM, the survival rate was 109.616%; when the concentration was 6.25 μM, the survival rate was 110.236%; when the concentration was 12.5 μM, the survival rate was 101.260%; when the concentration was 25 μM, the survival rate was 106.090%; when the concentration was 50 μM, the survival rate was 49.165%; when the concentration was 100 μM, the survival rate was 8.176%.

[0320] The above results show that the maximum safe concentration of I-4-6 for use in C2C12 muscle cells is 100μM or above; the maximum safe concentration of II-6-2 for use in C2C12 muscle cells does not exceed 25μM; the maximum safe concentration of II-8-1 for use in C2C12 muscle cells does not exceed 50μM; the maximum safe concentration of II-8-2 for use in C2C12 muscle cells does not exceed 25μM.

[0321] Table 3 is the statistical results of the effects of I-4-6, II-6-2, II-8-1 and II-8-2 on the activity of C2C12 muscle cells in Example 7, corresponding to the attached Figures 55-58 .

[0322] Table 3. Survival rate of C2C12 cells under the action of different concentrations of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0323]

[0324]

[0325] Example 8 Experiment on Alleviating Mouse Myoblast (C2C12) Atrophy Induced by Mouse Colon Cancer Cell C26 Supernatant I-4-6, II-6-2, II-8-1 and II-8-2

[0326] The diameter of differentiated mouse myoblasts (C2C12) was measured using a diameter measurement method. The staining method used was hematoxylin-eosin (HE) staining. Hematoxylin, an alkaline, positively charged dye, readily binds ionically to the negatively charged, acidic deoxyribonucleic acid (DNA) in the cell nucleus, staining it blue. Eosin, an acidic dye, dissociates in water into negatively charged anions and readily binds to the positively charged amino groups of proteins in the cytoplasm, staining them red. After staining, the cells were imaged under a high-magnification microscope, and the diameters of the myotubes were calculated using Digimizer software.

[0327] Western blotting was used to evaluate changes in protein levels in mouse myoblasts (C2C12). Protein samples separated by PAGE (polyacrylamide gel electrophoresis) were transferred to a solid support (e.g., nitrocellulose membrane). The solid support non-covalently adsorbed the proteins and maintained the identity and biological activity of the separated peptides. The proteins or peptides on the solid support served as antigens, reacted with corresponding antibodies, and then reacted with enzyme- or isotope-labeled secondary antibodies. Protein components expressed by the specific target gene separated by electrophoresis were detected by substrate development or autoradiography. This method was used to evaluate the effects of carnosol analogs on protein levels in a C2C12 cell model of muscular dystrophy.

[0328] The specific method is as follows: C2C12 cells were seeded at 30,000 cells / mL in a 24-well plate and differentiated into mature myotubes using high-glucose DMEM medium supplemented with 2% HS and 1% PS. Separately, C26 cells and 3T3-L1 adipocytes were seeded in T75 flasks and cultured in high-glucose DMEM supplemented with 10% FBS and 1% PS in a 5% CO2, 37°C cell culture incubator. Six million cells were passaged per flask. After 48 hours of incubation in 20 mL of culture medium, the supernatant was collected and centrifuged at 1000 rpm for 3 minutes. The supernatant was then collected and centrifuged at 4000 rpm for 10 minutes. The 3T3-L1 supernatant was mixed with 2% HS differentiation medium in a 1:1 ratio to serve as the healthy control group. The C26 supernatant was mixed with 2% HS differentiation medium in a 1:1 ratio to serve as the muscle atrophy-inducing medium. Equal volumes of the muscle atrophy-inducing medium were added to both the model and treatment groups. At the same time, I-4-6 stock solution was added to the cells at the following concentrations: 6.25 μM, 12.5 μM, and 25 μM (see Table 4); II-6-2 stock solution was added at 6.25 μM, 12.5 μM, and 25 μM (see Table 4); II-8-1 stock solution was added at 3.125 μM, 6.25 μM, 12.5 μM, and 25 μM (see Table 4); and II-8-2 stock solution was added at 3.125 μM, 6.25 μM, 12.5 μM, and 25 μM (see Table 4).

[0329] Table 4 shows the dosage regimen of I-4-6, II-6-2, II-8-1 and II-8-2 in the experiment of reversing muscle atrophy in C2C12 muscle cells in Example 8.

[0330] Table 4. Samples administered for the myoblast (C2C12) atrophy experiment in Example 8

[0331]

[0332]

[0333] Here, μM refers to μmol / L.

[0334] The myotube diameter was measured as follows:

[0335] After 48 hours of drug treatment, the cells were fixed with a fixative (anhydrous ethanol: formaldehyde: glacial acetic acid = 20:2:1) for more than 1 hour, stained with hematoxylin-eosin, and images were collected under a high-power microscope. Digimizer was used to calculate the myotube diameter.

[0336] The muscle atrophy reversal rate was calculated according to the following formula:

[0337] Muscle atrophy reversal rate = (average value of myotubes in the drug group - average value of myotubes in the model group) / (average value of myotubes in the control group - average value of myotubes in the model group) × 100%

[0338] The protein immunoblotting method is as follows:

[0339] Protein sample preparation: After 48 hours of drug exposure, remove the cell culture medium from the 6-well plate and wash three times with 37°C preheated PBS. After complete digestion with trypsin, terminate the digestion with culture medium containing 10% FBS + 1% PS. Centrifuge the cell suspension at 4°C, 1000 rpm for 5 minutes, wash once with ice-cold PBS, discard the supernatant, and centrifuge again for 3 minutes, discarding the supernatant. Add RIPA solution containing 1% phosphatase inhibitors as appropriate, vortex every 10 minutes, and lyse for a total of 30 minutes. Perform the entire lysis process on wet ice. After lysis, the lysate was centrifuged at 4°C and 12,000 rpm for 10 min. The supernatant was removed and the protein supernatant was diluted 5-fold (i.e., 5 μL of protein supernatant was added to 20 μL of ultrapure water). Protein standards at concentrations of 2 mg / mL, 1 mg / mL, 0.5 mg / mL, 0.25 mg / mL, 0.125 mg / mL, 0.0625 mg / mL, and 0.03125 mg / mL were prepared using ultrapure water. An ultrapure water control was set as a zero well, and the total volume was also 25 μL. BCA working solution was prepared at a ratio of solution A: solution B = 50:1. 200 μL and 25 μL of protein diluent were added to each group. The reaction was carried out in a 37°C oven in the dark for 30 min. The OD value was then measured at 562 nm on a microplate reader, and the protein concentration of the sample was calculated using the standard curve. The loading amount was set at 20 μg, the loading system was 20 μL, the protein was divided into packages according to the protein concentration and dilution multiple, heated in a 100°C metal bath for denaturation for 10 min, cooled on ice, and stored at -80°C after centrifugation or loaded directly.

[0340] Protein electrophoresis: Prepare the appropriate gel concentration and marker based on the target molecular weight. After the gel has solidified, place it in an electrophoresis tank filled with 1x running buffer. Remove the comb and remove any bubbles before loading the samples in order. Vortex the samples thoroughly before loading. Run the gel at 70V until the samples are flattened and the markers begin to separate. Then, reduce the voltage to 110V until the 25kD marker runs off the bottom edge of the gel.

[0341] Transfer: Prepare an appropriate amount of transfer buffer. Activate an appropriately sized PVDF membrane in anhydrous methanol for 30 seconds, then place the membrane in the prepared transfer buffer. Gently pry the two glass plates apart, trimming any unused areas and carefully removing the glue. Following the black glue and white film sandwich pattern, sandwich the membrane in the following order: black-surfaced plywood, sponge, three layers of filter paper, glue, PVDF membrane, three layers of filter paper, sponge, and white-surfaced plywood. Expel air bubbles at each step. Place the membrane in the electrophoresis tank and power on at 250mA for 1 hour on ice.

[0342] Immunofluorescence: After transfer, cut the PVDF membrane according to the target band location and place it in 5% milk in TPBS. Block the membrane on a shaker at room temperature for at least 1 hour. Remove the blocking solution and wash the membrane three times with TPBS for 10 minutes each. Prepare a primary antibody diluent in 5% milk in TPBS at a 1:1000 ratio and incubate with 1 mL overnight at 4°C. Remove the primary antibody and wash the membrane three times with TPBS for 10 minutes each. Prepare a secondary antibody diluent in 5% milk in TPBS at a 1:5000 ratio and incubate the membrane at room temperature for 1-2 hours on a shaker at room temperature. Wash the membrane three times with TPBS for 10 minutes each. Mix the ECL reagents A and B in a 1:1 ratio and apply evenly to the target protein. After the reaction is complete, place the membrane in an Amersham Imager 600 for chemiluminescence detection and photography.

[0343] Results and conclusions: Please refer to the attached Figures 59-78 Attached Figure 59-76 The following are representative HE staining images showing that Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 respectively alleviated the atrophy of C2C12 mature myotube cells induced by C26 cell culture medium. Figure 77 Figure 78 shows the myotube statistical results, and Appendix 6 shows the myotube statistical results. I-4-6, II-6-2, II-8-1, and II-8-2 showed significant reversal effects on myocyte atrophy in a concentration-dependent manner. At a concentration of 6.25 μM, I-4-6 achieved a reversal rate of 21.53%; at a concentration of 12.5 μM, the reversal rate was 61.39%; and at a concentration of 25 μM, the reversal rate was 67.08%. At a concentration of 6.25 μM, II-6-2 achieved a reversal rate of 28.40%; at a concentration of 12.5 μM, the reversal rate was 43.02%; and at a concentration of 25 μM, the reversal rate was 66.55%. When the concentration of Ⅱ-8-1 was 3.125 μM, the reversal rate was 42.17%; at 6.25 μM, the reversal rate was 35.55%; at 12.5 μM, the reversal rate was 42.63%; at 25 μM, the reversal rate was 73.21%. When the concentration of Ⅱ-8-2 was 3.125 μM, the reversal rate was 44.05%; at 6.25 μM, the reversal rate was 58.19%; at 12.5 μM, the reversal rate was 63.15%; and at 25 μM, the reversal rate was 88.52%.

[0344] Table 5 shows the statistical results of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 in alleviating C2C12 muscle cell atrophy, corresponding to the attached Figures 59-78 .

[0345] Table 5. Reversal rate of muscle atrophy in C2C12 myotube cells under the action of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0346] Carnosol derivatives (concentration) Muscle atrophy reversal rate (%) Ⅰ-4-6 (6.25 μM) 21.53 Ⅰ-4-6 (12.5 μM) 61.39 Ⅰ-4-6 (25 μM) 67.08 Ⅱ-6-2 (6.25 μM) 19.26 Ⅱ-6-2 (12.5 μM) 43.02 Ⅱ-6-2 (25 μM) 66.55 Ⅱ-8-1 (3.125 μM) 42.17 Ⅱ-8-1 (6.25 μM) 35.55 Ⅱ-8-1(12.5μM) 42.63 Ⅱ-8-1(25μM) 73.21 Ⅱ-8-2 (3.125 μM) 44.05 Ⅱ-8-2 (6.25 μM) 58.19 Ⅱ-8-2 (12.5 μM) 63.15 Ⅱ-8-2 (25 μM) 88.52

[0347] Attachment Figures 79-82 Immunoblotting results show the effects of I-4-6, II-6-2, II-8-1, and II-8-2 on protein levels in a C2C12 cell model of muscular dystrophy. The results show that C26 supernatant not only activates p-p65 in C2C12 myotubes, leading to overexpression of the E3 ubiquitin ligase Atrogin-1 and reduced expression of MHC and MyoD, but also inhibits AKT phosphorylation. I-4-6, II-8-1, or II-8-2 not only inhibits p-p65 expression in a concentration-dependent manner, thereby suppressing Atrogin-1 overexpression and promoting MHC and MyoD expression, but also inhibits the reduction in AKT phosphorylation induced by C26 supernatant in a concentration-dependent manner. On the one hand, Ⅱ-6-2 can not only inhibit the overexpression of Atrogin-1 and promote the expression of MHC, MyoD and MyoG in a concentration-dependent manner; at the same time, Ⅱ-6-2 can also upregulate the reduced AKT phosphorylation level caused by C26 supernatant in a concentration-dependent manner.

[0348] The above results indicate that Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 alleviate protein degradation in muscle cells and promote muscle cell differentiation and growth through a new molecular mechanism of action, and have a significant alleviating effect on muscle cell atrophy in a concentration-dependent manner.

[0349] Example 9 Experiment on the Effect of I-4-6, II-6-2, II-8-1 and II-8-2 on the Viability of Mouse Preadipocytes (3T3-L1)

[0350] The MTT assay was used to assess the viability of 3T3-L1 cells. The assay works by reducing exogenous MTT to water-insoluble, blue-purple formazan crystals in viable cells, while dead cells do not. A triple solution (10% SDS, 5% isobutanol, and 0.01% concentrated hydrochloric acid in water) dissolves the formazan in the cells. The absorbance at 570 nm is measured using an enzyme-linked immunosorbent assay (ELISA), indirectly reflecting the number of viable cells. Within a certain cell population range, the amount of MTT crystals formed is proportional to the cell number.

[0351] The above method can be used to evaluate the effect of drugs on the viability of mouse preadipocytes (3T3-L1). The specific method is as follows:

[0352] 3T3-L1 cells were seeded at 30,000 / mL in a 24-well plate and cultured in high-glucose DMEM supplemented with 10% FBS and 1% PS in a 5% CO2, 37°C incubator. After confluency and 6 days of incubation, cells were differentiated into mature adipocytes using 0.5 mM IBMX, 5 mg / mL insulin, 1 μM dexamethasone, and 10% FBS in high-glucose DMEM. Successful differentiation revealed numerous intracellular oil droplets. I-4-6, II-6-2, II-8-1, or II-8-2 were added to the differentiated 3T3-L1 adipocytes at the following concentrations: 1.0625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, or 100 μM in high-glucose DMEM supplemented with 10% FBS and incubated for 48 hours. The culture medium was replaced with a solution of MTT (5 mg / mL): culture medium = 1:10, 200 μL / well. After 4 hours, 50 μL of the triplicate solution was added to each well and incubated overnight. After overnight, the absorbance was measured at a wavelength of 570 nm using a microplate reader.

[0353] Cell viability (%) = (OD drug -OD blank ) / (OD control -OD blank )×100%

[0354] Results and conclusions: Please refer to the attached Figures 83-86 Attached Figures 83-86Table 7 shows the survival rates of 3T3-L1 cells treated with different concentrations of I-4-6, II-6-2, II-8-1, and II-8-2 for 48 hours. As shown in Table 7, when the I-4-6 concentration was 1.0625 μM, the survival rate was 95.898%; at 3.125 μM, the survival rate was 98.395%; at 6.25 μM, the survival rate was 101.607%; at 12.5 μM, the survival rate was 104.963%; at 25 μM, the survival rate was 94.231%; at 50 μM, the survival rate was 91.439%; and at 100 μM, the survival rate was 99.025%. When the concentration of Ⅱ-6-2 was 1.0625 μM, the survival rate was 107.97%; when the concentration was 3.125 μM, the survival rate was 113.401%; when the concentration was 6.25 μM, the survival rate was 115.701%; when the concentration was 12.5 μM, the survival rate was 95.76%; when the concentration was 25 μM, the survival rate was 100.736%; when the concentration was 50 μM, the survival rate was 108.566%; when the concentration was 100 μM, the survival rate was 99.726%. When the concentration of Ⅱ-8-1 was 3.125 μM, the survival rate was 104.226%; at 6.25 μM, the survival rate was 94.599%; at 12.5 μM, the survival rate was 95.514%; at 25 μM, the survival rate was 92.752%; at 50 μM, the survival rate was 92.876%; and at 100 μM, the survival rate was 71.851%. When the concentration of Ⅱ-8-2 was 3.125 μM, the survival rate was 100.707%; at 6.25 μM, the survival rate was 103.165%; at 12.5 μM, the survival rate was 96.889%; at 25 μM, the survival rate was 97.478%; at 50 μM, the survival rate was 92.876%; and at 100 μM, the survival rate was 80.929%.

[0355] The above results show that the maximum safe concentration of Ⅰ-4-6 for 3T3-L1 adipocytes is 100μM or above; the maximum safe concentration of Ⅱ-6-2 for 3T3-L1 adipocytes is 100μM or above; the maximum safe concentration of Ⅱ-8-1 for 3T3-L1 adipocytes does not exceed 50μM; the maximum safe concentration of Ⅱ-8-2 for 3T3-L1 adipocytes does not exceed 50μM.

[0356] Table 6 is the statistical results of the toxicity experiments of I-4-6, II-6-2, II-8-1 and II-8-2 on 3T3-L1 adipocytes in Example 9, corresponding to the appended Figures 83-86 .

[0357] Table 6. Survival rate of 3T3-L1 cells under different concentrations of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0358]

[0359]

[0360] Example 10 Experimental Results of I-4-6 and II-6-2 Alleviating Lipolysis in 3T3-L1 Adipocytes

[0361] Oil Red O fat staining was used to evaluate intracellular fat content. Oil Red O is a fat-soluble dye that is highly soluble in fat and can specifically stain neutral fats such as triglycerides in tissues.

[0362] Western blotting was used to evaluate the changes in the levels of related proteins in mouse adipocytes (3T3-L1).

[0363] The above method can be used to evaluate the effects of drugs on a cellular lipolysis model. Specifically, 3T3-L1 cells were seeded at 30,000 cells / mL in a 24-well plate and cultured in high-glucose DMEM supplemented with 10% FBS and 1% PS in a 5% CO2, 37°C cell culture incubator. After the cells reached confluency and continued to be confluent for 6 days, they were differentiated into mature adipocytes using high-glucose DMEM supplemented with 0.5 mM IBMX, 5 mg / mL insulin, 1 μM dexamethasone, and 10% FBS. After successful differentiation, numerous oil droplets were clearly visible within the cells. Separately, C26 cells and C2C12 myotubes were seeded in T75 flasks and cultured in fresh phenol red-free high-glucose DMEM after confluency. After 48 hours, the C26 cell supernatant in phenol red-free high-glucose DMEM was collected and centrifuged at 1000 rpm for 3 minutes, followed by centrifugation at 4000 rpm for 10 minutes. The C2C12 cell supernatant and phenol red-free, high-glucose DMEM culture medium were mixed in a 1:1 ratio to serve as the healthy control group. The C26 cell supernatant and phenol red-free, high-glucose DMEM culture medium were mixed in a 1:1 ratio to serve as the lipolysis-inducing solution. Equal amounts of the lipolysis-inducing solution were added to both the model and treatment groups. Simultaneously, I-4-6 stock solution was added to the cells at the following concentrations: 6.25 μM, 12.5 μM, and 25 μM (see Table 7). II-6-2 stock solution was added to the cells at the following concentrations: 6.25 μM, 12.5 μM, and 25 μM (see Table 7).

[0364] Table 7 shows the dosage regimen of I-4-6 in the experiment of reversing lipolysis in 3T3-L1 adipocytes in Example 10.

[0365] Table 7. Samples administered for the mouse preadipocyte (3T3-L1) lipolysis experiment in Example 10

[0366]

[0367] The Oil Red O staining and semi-quantitative method is as follows: After 48 hours of drug action, dilute 0.5% Oil Red O dye (prepared with isopropanol) with distilled water in a ratio of 3:2, mix well, and filter with a 0.42μm filter head as a staining agent. Discard the cell culture medium, fix the cells with 4% neutral formaldehyde for more than 1 hour, use a vacuum pump to remove the fixative, wash with PBS 3 times, place in a ventilated place at room temperature to dry for 20 minutes, add Oil Red O staining solution and stain for 30 minutes, then remove it, wash with PBS 2 times, and then use 60% isopropanol to wash off the floating color, wash with distilled water 2 times, and observe under an inverted high-power microscope and take pictures. After that, use isopropanol to dissolve the Oil Red O bound to the cells, and use the undifferentiated adipocyte staining group to perform a parallel operation as a blank zeroing group. The staining group is also performed in parallel as a blank zeroing group, and the OD value is measured at 510nm with an enzyme reader. According to the OD value results, the Oil Red O semi-quantification is performed according to the following formula:

[0368] Fat content (%) = (OD drug -OD blank ) / (OD control -OD blank )×100%

[0369] The protein immunoblotting method was the same as described in Example 8.

[0370] Results and conclusions: Please refer to the attached Figures 87-98 Attached Figures 87-96 The following are representative images of Oil Red O staining showing that Ⅰ-4-6 and Ⅱ-6-2 alleviate C26 supernatant-induced lipolysis in 3T3-L1 mature adipocytes. In the normal control group, a large number of lipid droplets accumulated in the cells, and the lipid droplets were large, darkly stained, and brightly colored. After the action of C26 supernatant alone, the intracellular lipid droplets significantly became smaller and the content decreased sharply. When C26 supernatant was incubated simultaneously with -4-6 or Ⅱ-6-2, as the concentration of Ⅰ-4-6 or Ⅱ-6-2 increased, the content of Oil Red O-stained fat droplets in the cells gradually increased and the lipid droplets became larger.

[0371] Attachment Figure 97 , 98 is the semi-quantitative result of Oil Red O staining, such as Figure 97 As shown in Figure 98, I-4-6 and II-6-2 increased the intracellular oil red O content in a concentration-dependent manner, indicating an increase in fat content.

[0372] Attachment Figure 99The results of a western blot experiment using I-4-6 in a 3T3-L1 cell lipolysis model were shown. The results showed that under stimulation with C26 supernatant, p-p65 in 3T3-L1 adipocytes was activated and its expression was upregulated. I-4-6 reduced p-p65 expression in a concentration-dependent manner. The C26 cell culture medium activated the phosphorylation of HSL (hormone-sensitive esterase), upregulating p-HSL expression and contributing to lipolysis. As the I-4-6 concentration increased, p-HSL expression decreased. The C26 cell culture medium activated the phosphorylation of AMPKα (AMP-dependent protein kinase α isoform), upregulating p-AMPKα expression and enhancing energy metabolism. As the I-4-6 concentration increased, p-AMPKα expression decreased.

[0373] The above results indicate that Ⅰ-4-6 reduces fat degradation and alleviates excessive energy consumption through a new molecular mechanism of action, and has a significant alleviating effect on adipocyte lipolysis in a concentration-dependent manner.

[0374] Example 11 PK experiments of I-5-3, I-4-22, I-4-6, II-6-2 and II-8-1

[0375] BALB / c mice were injected with Ⅰ-5-3, Ⅰ-4-22, Ⅰ-4-6, Ⅱ-6-2 or Ⅱ-8-1 via tail vein. The solvent was 5% DMSO + 1% HS-15 + 94% 0.9% Saline. The dose was 25 mg / kg.

[0376] Results and conclusions: Please refer to the attached Figure 100 and Table 8. AUC of I-5-3, I-4-22, I-4-6, II-6-2, and II-8-1 0-t (ng·hr / mL), T 1 / 2 The blood drug curve area (AUC) of I-4-6 and II-6-2 were 1907 ng·hr / mL and 1.01 h, 520 ng·hr / mL and 0.98 h, 8369 ng·hr / mL and 11.1 h, 3711 ng·hr / mL and 8.8 h, and 273 ng·hr / mL and 1.03 h, respectively. 0-t ), drug half-life (T 1 / 2 ) and drug retention time (MRT) are ideal.

[0377] Table 8 is the PK test results of I-4-6, II-6-2, II-8-1 and II-8-2 in Example 11, corresponding to the attached Figure 100 Table 8. Parameters of PK experiments of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0378]

[0379] Example 12 Experimental results of treating tumor cachexia animal model with I-4-6 and II-6-2

[0380] C26 cell suspension was pre-inoculated into BALB / c mice in the left and right armpits at 1.5 million cells each. 3 The tumor was removed at 4 ℃ and homogenized with 0.5 mL PBS / g to obtain a tumor tissue suspension, which was counted and diluted to 1 million / 100 μL. The mice to be inoculated were grouped according to their weight, and the cell suspension was inoculated into the left armpit of BALB / c mice with an inoculation volume of 100 μL / mouse. I-4-6 and II-6-2 were mixed with 3% DMSO, preheated 2% Ethanol, 1% HS-15 and 94% 0.9% Saline solution to form a uniform and stable solution with a final concentration of 4 mg / mL. The dosage was 40 mg / kg and the administration route was intraperitoneal injection. The drug was administered on the third day after inoculation. The weight, body temperature, tumor size and food intake of the mice were monitored every day. On the 16th day, the body weight of the mice in the model group decreased by about 15% or the tumor volume reached 2000 mm 3 When the number of cells in the lungs increases by about 1%, the patient is considered to have entered the late stage of cachexia. Mice were sacrificed by cervical dislocation and biochemical samples of gastrocnemius muscle, epididymal fat, tumor, and serum were obtained.

[0381] Attachment Figures 101-105 Figures show the tumor-bearing body weight, tumor-free body weight, tumor volume, tumor weight, and tumor anatomy during the mice's survival period. As shown, the weight of mice in the healthy group continued to increase; the tumor-bearing body weight of mice in groups I-4-6, II-6-2, and the C26 tumor model group continued to rise from the start of the experiment to day 11, then began to decline sharply from day 11 until the end of the experiment, as did the tumor-free body weight. In contrast, the tumor-bearing and tumor-free body weights of the I-4-6 group decreased more gradually, and from days 14 to 16, both tumor-bearing and tumor-free body weights were higher than those of the C26 model group, with statistically significant differences (p < 0.5).

[0382] Attachment Figures 106-107 The gastrocnemius muscle mass of mice and the actual photos of the gastrocnemius muscle show that the gastrocnemius muscle mass of groups Ⅰ-4-6 is no different from that of the C26 tumor model group, and there is no statistical significance in the mass.

[0383] Attachment Figures 108-109 The epididymal fat mass of mice and the actual photos of epididymal fat show that the epididymal fat mass of epididymal fat group Ⅰ-4-6 is significantly greater than that of C26 tumor model group, and the difference is statistically significant (p<0.5).

[0384] These results indicate that Ⅱ-6-2 cannot alleviate the weight loss, muscle atrophy, and fat degradation caused by tumor cachexia without affecting tumor size. Ⅰ-4-6 can alleviate the weight loss caused by tumor cachexia by inhibiting fat degradation without affecting tumor size.

[0385] The above is a preferred embodiment of the present invention. It should be pointed out that ordinary technicians in the technical field covered by the present invention can make several supplements and improvements without departing from the method of the present invention, but these supplements and improvements should also be regarded as the scope of protection of the present invention.

Claims

1. A carnols compound, characterized in that: The invention includes a compound represented by formula (Y) or a pharmaceutically acceptable salt or optical isomer of a compound represented by formula (Y), wherein the formula (Y) is as shown below: In the formula (Y), A is N or O; D is selected from OH, OTf, methoxy, substituted or unsubstituted benzene ring; E is selected from OH, OTf, methoxy, substituted or unsubstituted benzene ring; the substituent is halogen or methoxy; R2 is selected from H, substituted or unsubstituted C1-C 12 Alkyl; the substituent is selected from halogen, trifluoromethyl, 5-7 membered aromatic heterocycle, 3-6 membered ring, 6 membered heterocyclic group, condensed ring, substituted or unsubstituted benzene ring; the substituent of the benzene ring is halogen or trifluoromethyl; the aromatic heterocyclic group and 6 membered heterocyclic group contain one or more heteroatoms selected from N, O, S; R3 is selected from OH or a substituted aliphatic group; the substituent is selected from phenyl and a 6-membered aromatic heterocyclic group; the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, and S.

2. Use of the carnosol compound according to claim 1 in the preparation of a medicament for treating cachexia.

3. A pharmaceutical composition, characterized in that The composition contains the carnosin compound according to claim 1 and / or further contains other pharmaceutically acceptable ingredients.

4. Use of the pharmaceutical composition according to claim 3 in preparing a pharmaceutical composition for treating tumor cachexia.

5. The use according to claim 4, characterized in that The product dosage form of the pharmaceutical composition for treating tumor cachexia is capsule, tablet, oral preparation, microcapsule preparation, injection, ointment, spray or suppository; the administration method of the product for treating cachexia is injection, oral, parenteral, inhalation spray or transdermal administration.

6. The use according to claim 4 or 5, characterized in that The tumor cachexia is muscle atrophy caused by tumor tissue; and / or, the tumor cachexia is fat loss caused by tumor tissue; and / or, the tumor cachexia is decreased appetite caused by tumor tissue.

7. The use according to claim 6, characterized in that The tumor is a solid tumor.

8. The use according to claim 6, characterized in that The tumor cachexia includes tumor cachexia caused by digestive tract related cancers, liver cancer and lung cancer.

9. The use according to claim 5, 7 or 8, characterized in that The carnosol compound of formula (Y) accounts for 0.001-100 wt.% of the total dry weight of the composition.

10. Use of the carnosol compound according to claim 1 in health care products, characterized in that: The health care products include protein source, fat source and / or carbohydrate source products; selected from nutritionally balanced foods, complete nutritional formulas, dairy products, frozen or room temperature stable beverages, soups, nutritional bars, desserts, pet foods, pharmaceutical compositions or combinations thereof.

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