Salvianolic acid derivative and its application in drugs for treating cancer cachexia

Abstract

The present invention belongs to the field of pharmaceutical synthesis, and specifically relates to salviphorol derivatives and their application in drugs for treating cancer cachexia. Salviphorol compounds include compounds represented by formula (X) and / or formula (Y), or pharmaceutically acceptable salts or optical isomers of the compounds represented by formula (X) and / or formula (Y). The present invention also provides a pharmaceutical composition for treating cachexia diseases, and its application in the preparation of drugs for treating cancer cachexia, including muscle atrophy caused by tumor tissues, fat reduction caused by tumor tissues, reduced appetite caused by tumor tissues, inflammatory responses caused by tumor tissues, and cancer cachexia caused by digestive tract-related cancers, liver cancer, lung cancer, and colon cancer. The present invention also provides its application in the preparation of drugs or inhibitors for treating or inhibiting muscle atrophy, drugs or inhibitors for inhibiting or alleviating adipocyte lipolysis, and drugs or inhibitors for inhibiting or alleviating weight loss or reduction.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical synthesis, and specifically relates to salviphenol derivatives and their application in drugs for treating cancer cachexia. Background Art

[0002] Cancer cachexia is a serious complication caused by tumors, which is a multi-factor syndrome characterized by continuous loss of skeletal muscle (with or without fat), anorexia, anemia and involuntary weight loss, and involves metabolic changes in various tissues and organs. Skeletal muscle depletion is a major feature of cancer cachexia, and its consequences include increased chemotherapy toxicity, cancer surgery complications and mortality. Cancer cachexia is highly correlated with pancreatic cancer, esophageal cancer, gastric cancer, lung cancer, liver cancer and colon cancer, causing about half of cancer deaths worldwide, and it is more common in advanced cancer patients. The prevalence of cachexia in cancer patients is as high as 50%-80%. The prevalence of cancer cachexia varies depending on the type of tumor. Among them, the prevalence in breast cancer and leukemia patients is about 40%, in lung cancer patients is 30%, in colon cancer and prostate cancer patients is about 50%, in liver cancer patients is about 41%-45%, in pancreatic cancer patients is about 70%, and in gastric cancer patients is about 80%-87%. The frequency of weight loss in pancreatic cancer or gastric cancer patients is the highest, exceeding 60%-80%, while the incidence of weight loss in lung cancer, colorectal cancer or prostate cancer patients is more than 50%. Cancer cachexia not only seriously affects the effects of chemotherapy, radiotherapy and drug treatment, but also damages the quality of life of patients and shortens their survival life. Seeking methods, targets or drugs for treating cancer cachexia to treat cancer cachexia patients is extremely urgent.

[0003] The currently known pathogenic mechanisms of cancer cachexia are mainly closely related to systemic inflammation, skeletal muscle loss and atrophy, fat loss, anorexia, multi-organ syndrome and energy metabolism disorders, and there are no specific treatment methods and drugs. Currently, non-drug palliative treatment methods are mostly used clinically, such as nutritional supplementation, exercise therapy, etc. However, the weight loss caused by cancer cachexia is different from that caused by malnutrition. In addition to the consumption of adipose tissue, more importantly, there is atrophy of skeletal muscle, the largest organ of the human body. Moreover, not only will the synthesis of muscle protein decrease, but there will also be an increase in muscle tissue degradation. Therefore, nutritional supplementation can only relieve the weight loss caused by cachexia to a certain extent and cannot solve the pathological problems of cachexia. And exercise is difficult to achieve for patients who have developed cancer cachexia and have muscle atrophy. Therefore, the current palliative treatment methods can only relieve the symptoms of cancer cachexia to a certain extent and are difficult to achieve the purpose of prevention and treatment.

[0004] Alamoline is an orally administered small molecule (583.2 g / mol) ghrelin receptor agonist, which is considered to improve cancer cachexia by increasing appetite, showed excellent results in recent Phase III studies, and obtained regulatory approval for the treatment of cancer cachexia in Japan. Medroxyprogesterone acetate is a synthetic progesterone and appetite stimulant, which has been approved for the treatment of anorexia, cachexia or weight loss of unknown cause. Recently, Ruiz-Garcia et al. reviewed the clinical trials of medroxyprogesterone acetate and found that although medroxyprogesterone acetate can cause weight gain, it has no effect on improving quality of life.

[0005] In summary, although an endless stream of drugs or antibodies for the treatment of cancer cachexia have been developed in recent years, it is very difficult to continue due to poor clinical effects. Chinese herbal medicine has a history of thousands of years in the treatment of chronic wasting diseases, providing an important direction for the study of the treatment of cancer cachexia.

[0006] Salvigenol is a natural active ortho-diphenol diterpenoid compound, one of the active ingredients of plants such as rosemary and sage, with the molecular formula 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 salvigenol can inhibit the proliferation of tumor cells in diseases such as breast cancer, colon cancer, and prostate cancer. In addition, salvigenol has been identified as an effective antioxidant and is involved in the treatment of various diseases. Previously, we found that salvigenol has a good anti-cancer cachexia effect and applied for a patent for "the application of salvigenol compounds in the preparation of drugs for the treatment of cachexia diseases", with the patent number ZL 202010089693.1. However, the structure of salvigenol in this application is single and cannot meet the current needs. Summary of the Invention

[0007] The object of the present invention is to address the problems existing in the prior art. On this basis, we have carried out structural modification on carnosol and provided a synthesis of carnosol derivatives, such as carnosol analogues with new structures, and studied their effects in alleviating muscle atrophy and lipolysis caused by cancer cachexia. In terms of muscle, on the one hand, it inhibits the expression of p-p65 in a concentration-dependent manner, thereby inhibiting the overexpression of Atrogin-1 and playing a role in alleviating muscle cell atrophy; on the other hand, carnosol derivatives up-regulate the phosphorylation level of AKT in a concentration-dependent manner, promote the expression of MHC, MyoD, and MyoG, and accelerate the synthesis of myotubular proteins. 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 gradient manner and weaken the excessive lipolysis of 3T3-L1 adipocytes; on the other hand, carnosol derivatives inhibit the up-regulation of the expression level of p-AMPKα in 3T3-L1 adipocytes in a concentration-dependent manner, avoiding excessive energy consumption and loss of fat. In short, in vitro, carnosol derivatives can reduce protein degradation in muscle cells, promote protein synthesis and myocyte growth and differentiation in muscle cells; reduce lipolysis and excessive energy consumption in adipocytes; in vivo, carnosol derivatives can alleviate the degradation of adipose tissue caused by cancer cachexia, reduce weight loss, and play a role in alleviating the symptoms of cancer cachexia, and are candidate compounds with great prospects for treating cancer cachexia.

[0008] To achieve the above object of the invention, the technical solution of the present application is as follows:

[0009] 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),

[0010]

[0011] In the said formula (X),

[0012] A is O or N; B is O or N; C is N;

[0013] When A is O, R1 is selected from H or a substituted or unsubstituted 5-7-membered aromatic heterocyclic group or phenyl or a substituted or unsubstituted C1-C 12 alkyl group, and the substituents are selected from one or more of halogen, C1-C6 alkyl, cyano, trifluoromethyl, nitro, carboxyl, hydroxyl, hydroxymethyl, methoxy, methyl, amino, acetylamino, methanesulfonyl, and methoxyamide group, and the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, and S;

[0014] When A is N, R1 is selected from H or a substituted or unsubstituted 5-7-membered aromatic heterocyclic group or phenyl or a substituted or unsubstituted C1-C 12 alkyl group, and the substituents are selected from one or more of halogen, C1-C6 alkyl, cyano, trifluoromethyl, nitro, carboxyl, hydroxyl, hydroxymethyl, methoxy, methyl, amino, acetamido, methanesulfonyl, methoxyamido; the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, S; R2 is selected from H or a substituted or unsubstituted C1-C 12 alkyl group, and the substituents are selected from halogen, trifluoromethyl, 5-7-membered aromatic heterocyclic group, 3-6-membered ring group, 6-membered heterocyclic group, fused ring group, substituted or unsubstituted phenyl, and the substituents of the phenyl are halogen, trifluoromethyl, and the aromatic heterocyclic group and 6-membered heterocyclic group contain one or more heteroatoms selected from N, O, S.

[0015] In particular, some compounds have the following isomers:

[0016]

[0017] The general formula (Y) is as follows:

[0018]

[0019] In the general formula (Y),

[0020] A is N or O; D is selected from OH or OTf or methoxy or a substituted or unsubstituted benzene ring; E is selected from OH or OTf or methoxy or a substituted or unsubstituted benzene ring, and the substituents are halogen, methoxy;

[0021] R2 is selected from H or a substituted or unsubstituted C1-C 12 alkyl group, and the substituents are selected from halogen, trifluoromethyl, 5-7-membered aromatic heterocycle, 3-6-membered ring, 6-membered heterocyclic group, fused ring, substituted or unsubstituted benzene ring, and the substituents of the benzene ring are halogen, trifluoromethyl, and the aromatic heterocyclic group and 6-membered heterocyclic group contain one or more heteroatoms selected from N, O, S;

[0022] R3 is selected from OH or a substituted fatty group, and the substituents are selected from phenyl, 6-membered aromatic heterocyclic group, and the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, S.

[0023] Preferably, A = O, B = O, C = N, R1 =

[0024] 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 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 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 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 provides a compound with a novel structure, namely, the structural formula of (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(methano)phenanthro[3,4-d]oxazol-12-one (Compound 1) is as follows:

[0041]

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

[0043] Furthermore, the drug is a drug applicable to humans and / or animals.

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

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

[0046] Even further, the administration method of the product for treating cachexia is injection, oral administration, parenteral administration, inhalation spray or transdermal administration.

[0047] Even further, the product for treating cachexia is a product for treating cancer cachexia.

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

[0049] Even further, the product contains the general formula (X) and / or general formula (Y) in an amount of 0.001-100 wt.-% of the total dry weight of the composition.

[0050] Even further, 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 diseases, wherein the dosage of (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(methano)phenanthro[3,4-d]oxazol-12-one (Compound 1) shown in formula (1) is 0.01 μg-100 mg / kg body weight / day.

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

[0053] Even further, the product is in the form selected from nutritionally balanced foods, total nutrient formulas, dairy products, frozen or ambient-stable beverages, soups, nutrition bars, confections, pet foods, pharmaceutical compositions and combinations thereof.

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

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

[0056]

[0057] The compound of formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(methano)phenanthro[3,4-d]oxazol-12-one (Compound 1) of the present invention can be used to prepare anti-cachexia drugs. Among them, the cachexia includes, but is not limited to, progressive wasting syndromes induced by tumors, chronic kidney diseases, chronic obstructive pulmonary diseases, chronic heart failure, AIDS, etc. The drugs are for human and / or animal use.

[0058] The term "sageol compounds" used in the present invention refers to a single component or a composition composed of other pharmaceutically acceptable components acting in combination.

[0059] The term "other pharmaceutically acceptable components" used in the present invention refers to drugs that have no antagonistic effect with general formula (X) and general formula (Y), and can also be any one or more pharmaceutically acceptable excipients.

[0060] The term "cachexia" used in the present invention refers to cachexia induced by solid tumors.

[0061] The term "solid tumor" used in the present invention refers to primary or secondary solid tumors such as digestive tract-related cancers, liver cancers, and lung cancers.

[0062] The muscle atrophy described in the present invention is muscle atrophy induced by tumor cachexia.

[0063] The fat degradation described in the present invention is fat degradation caused by tumor cachexia.

[0064] The administration routes of the general formula (X) and general formula (Y) sageol compounds or a composition composed of them and other pharmaceutically acceptable components are injection, oral administration, parenteral administration, inhalation spray, or transdermal administration.

[0065] In the present invention, the application of the salvianolic acid compounds of general formula (X) and general formula (Y) in the preparation of drugs for treating cachexia diseases, said drugs can be used for humans and / or animals. Among them, the dosage of the salvianolic acid compounds of general formula (X) and general formula (Y) is 0.01 μg - 100 mg / kg body weight / day. Among them, said drugs include drugs or inhibitors for treating or inhibiting muscle atrophy, drugs or inhibitors for inhibiting or alleviating lipolysis of adipocytes, drugs or inhibitors for inhibiting or alleviating weight loss or reduction.

[0066] The third object of the present application is to protect a pharmaceutical composition for treating cancer cachexia diseases, said composition contains the salvianolic acid compounds shown by general formula (X) and / or general formula (Y), and / or further contains other pharmaceutically acceptable ingredients

[0067] Furthermore, said other pharmaceutically acceptable ingredients can be drugs that have no antagonistic effect with the salvianolic acid compounds, or can be any one or more excipients permitted in pharmacy.

[0068] Among them, the product dosage forms of said composition include but are not limited to capsules, tablets, oral preparations, microcapsule preparations, injections, ointments, sprays or suppositories, etc. Among them, the administration methods of said composition include but are not limited to injection, oral administration, parenteral administration, inhalation spray or transdermal administration, etc. Among them, the salvianolic acid compounds shown by general formula (X) and / or general formula (Y) account for 0.001 - 100 wt.-% of the total dry weight of said composition.

[0069] In the application of the present invention, said cachexia diseases include cancer cachexia; said cancer is solid tumor. Among them, said cancer cachexia includes but is not limited to muscle atrophy caused by tumor tissues, fat reduction caused by tumor tissues, reduced appetite caused by tumor tissues, inflammatory reactions caused by tumor tissues, and cancer cachexia caused by digestive tract-related cancers, liver cancer, lung cancer, colon cancer, etc.

[0070] The present invention also proposes a method for treating cachexia diseases, the dosage of the salvianolic acid compounds shown by general formula (X) and general formula (Y) is 0.01 μg - 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, etc.

[0071] The present invention also proposes a pharmaceutical composition containing a therapeutically effective amount of the salvianolic acid compounds shown by general formula (X) and general formula (Y) and their medicinal salts.

[0072] The present invention also provides a pharmaceutical composition for treating cachectic diseases, said composition containing the compound of formula (1) (11aR)-2-(4-bromophenyl)-4-isopropyl-8,8-dimethyl-7,7a,8,9,10,11-hexahydro-6H-6,11a-(methano)phenanthro[3,4-d]oxazol-12-one, and / or other pharmaceutically acceptable ingredients. The product dosage forms of the pharmaceutical composition include but are not limited to capsules, tablets, oral preparations, microcapsule preparations, injections, ointments, sprays or suppositories, etc. The product administration methods of the pharmaceutical composition include but are not limited to injection, oral administration, parenteral administration, 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-(methano)phenanthro[3,4-d]oxazol-12-one shown in formula (1) accounts for 0.001-100 wt.-% of the total dry weight of the composition.

[0073] The present invention also provides that the composition product further includes a protein source, a fat source and / or a carbohydrate source, and may also be selected from the forms of nutritionally balanced foods, total nutrition formulas, dairy products, frozen or ambient-stable beverages, soups, nutrition bars, sweets, pet foods, pharmaceutical compositions and their combinations, etc. The product contains the salvianolic acid compounds shown in general formula (X) and general formula (Y).

[0074] The present invention also provides that the salvianolic acid compounds shown in general formula (X) and general formula (Y) include but are not limited to drugs for humans and / or pets, foods for humans and / or pets, nutritional supplements and / or health products.

[0075] The present invention also provides that the salvianolic acid compounds shown in general formula (X) and general formula (Y) are used as a single ingredient or combined with other pharmaceutically acceptable ingredients to form a composition for preparing a product for treating cachexia. The other pharmaceutically acceptable ingredients contained therein may be drugs that have no antagonistic effect with this compound, or any one or more excipients permitted in pharmacy.

[0076] Compared with the prior art, the present invention has the following beneficial effects:

[0077] We have carried out structural modification on carnosol. The research found that the newly modified carnosol derivatives (Carnosol analogues) in this patent have the effects of alleviating muscle atrophy and fat lipolysis caused by cancer cachexia. In terms of muscle, on the one hand, its mechanism inhibits the expression of p-p65 in a concentration-dependent manner, thereby inhibiting the overexpression of Atrogin-1, playing a role in alleviating muscle cell atrophy; on the other hand, the carnosol derivatives up-regulate the phosphorylation level of AKT in a concentration-dependent manner, promote the expression of MHC, MyoD and MyoG, and accelerate the synthesis of myotubular proteins. 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 gradient manner, and weaken the excessive lipolysis of 3T3-L1 adipocytes; on the other hand, the carnosol derivatives inhibit the up-regulation of the expression level of p-AMPKα in 3T3-L1 adipocytes in a concentration-dependent manner, avoiding excessive energy consumption and loss of fat. In short, in vitro, the carnosol derivatives can reduce the protein degradation in muscle cells, promote the protein synthesis and myocyte growth and differentiation in muscle cells; reduce the lipolysis of adipocytes and excessive energy consumption; in vivo, the carnosol derivatives can alleviate the degradation of adipose tissue caused by cancer cachexia, reduce weight loss, and play a role in alleviating the symptoms of cancer cachexia. It is a candidate compound with great prospects for the treatment of cancer cachexia. Description of the Drawings

[0078] Figure 1 It is the HE staining diagram of the control group in Table 2 of Example 6.

[0079] Figure 2 It is the HE staining diagram of the model group in Table 2 of Example 6.

[0080] Figures 3 - 54 They are respectively the HE staining diagrams of experimental groups 1-52 in Table 2 of Example 6.

[0081] Figure 55 -58 are respectively the experimental results of the effects of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, Ⅱ-8-2 on the activity of C2C12 muscle cells in Example 7.

[0082] Figure 59 It is the HE staining diagram of control group 1 in Table 4 of Example 8.

[0083] Figure 60 It is the HE staining diagram of model group 1 in Table 4 of Example 8.

[0084] Figure 61 -76 are respectively the HE staining diagrams of experimental groups 1-14 in Table 4 of Example 8.

[0085] Figure 77 It is the statistical result diagram of Ⅰ-4-6 and Ⅱ-6-2 in Example 8 alleviating C2C12 muscle cell atrophy.

[0086] Figure 78 It is the statistical result graph of Ⅱ-8-1 and Ⅱ-8-2 in Example 8 alleviating C2C12 muscle cell atrophy.

[0087] Figures 79 - 82 They are the results of the protein immunoblotting experiments of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 in the C2C12 cell muscle atrophy model in Example 8 respectively.

[0088] 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 respectively.

[0089] Figure 87 It is the Oil Red O staining graph of control group 1 in Table 7 of Example 10.

[0090] Figure 88 It is the Oil Red O staining graph of C26 model group 1 in Table 7 of Example 10.

[0091] Figures 89 - 91 It is the Oil Red O staining graph of experimental groups 1-3 in Table 7 of Example 10.

[0092] Figure 92 It is the Oil Red O staining graph of control group 2 in Table 7 of Example 10.

[0093] Figure 93 It is the Oil Red O staining graph of C26 model group 2 in Table 7 of Example 10.

[0094] Figure 94 -96 is the Oil Red O staining graph of experimental groups 4 -6 in Table 7 of Example 10.

[0095] Figures 97 - 98 They are the semi-quantitative result graphs of the Oil Red O staining experiments of Ⅰ-4-6 and Ⅱ-6-2 alleviating lipolysis of 3T3-L1 adipocytes in Example 10 respectively.

[0096] Figure 99 It is the result of the protein immunoblotting experiment of Ⅰ-4-6 in the 3T3-L1 cell lipolysis model in Example 10.

[0097] Figure 100 It is the PK experimental results of Ⅰ-5-3, Ⅰ-4-22, Ⅰ-4-6, Ⅱ-6-2, and Ⅱ-8-1 in Example 11.

[0098] Figure 101 It is the tumor-bearing body weight graph of experimental mice of Ⅰ-4-6 and Ⅱ-6-2 in Example 12.

[0099] Figure 102It is the graph of the tumor-free body weights of the experimental mice of Ⅰ-4-6 and Ⅱ-6-2 in Example 12.

[0100] Figure 103 It is the graph of the tumor volume changes of the experimental mice of Ⅰ-4-6 and Ⅱ-6-2 in Example 12.

[0101] Figure 104 It is the graph of the tumor weights of the experimental mice of Ⅰ-4-6 in Example 12.

[0102] Figure 105 It is the anatomical diagram of the tumors of the experimental mice of Ⅰ-4-6 in Example 12.

[0103] Figure 106 It is the graph of the gastrocnemius muscle masses of the experimental mice of Ⅰ-4-6 in Example 12.

[0104] Figure 107 It is the anatomical diagram of the gastrocnemius muscles of the experimental mice of Ⅰ-4-6 in Example 12.

[0105] Figure 108 It is the graph of the epididymal fat masses of the experimental mice of Ⅰ-4-6 in Example 12.

[0106] Figure 109 It is the anatomical diagram of the epididymal fat of the experimental mice of Ⅰ-4-6 in Example 12. Detailed implementation manners

[0107] The present invention will be further described below in conjunction with examples, but these examples are by no means any limitation to the present invention. In all examples, the purification of the final product was prepared and purified by the high performance liquid chromatograph LC-3000 of our research group at Sichuan University of Science & Engineering. The liquid phase column was Extend-C18, 0.5um, 30*100mm. 1 1H-NMR was measured by a Bruker AvanceIII 600MHz nuclear magnetic resonance spectrometer of Sichuan University of Science & Engineering, and the chemical shift was expressed in δ (ppm); the mass spectrometry was measured by an AB Sciex Triple TOF 5600 mass spectrometer of Kunming Institute of Botany.

[0108] The syntheses 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, tetrakis(triphenylphosphine)palladium, boric acid derivatives, sodium cyanoborohydride were purchased from Anhui Zesheng Technology Co., Ltd., sodium borohydride was purchased from Chengdu Kelong Chemical Co., Ltd., iodomethane, 2,2-dimethoxypropane, pyridinium p-toluenesulfonate, chromium trioxide, hydroxylamine hydrochloride, tetraisopropyl titanate, amine compounds, trifluoroacetic acid were purchased from Shanghai Titan Scientific Co., Ltd.

[0109] In the present invention, the pharmacodynamic test methods adopted are well-known and recognized by those skilled in the art of technology research and development.

[0110] In the present invention, the C2C12 cells (mouse myoblasts), 3T3-L1 cells (mouse adipocytes) and C26 cells (mouse colon cancer cells) were purchased from the Cell Bank of the Chinese Academy of Sciences' Committee for Type Culture Collection. BALB / c mice were purchased from Shanghai Jihui or Slake Co., Ltd.

[0111] Carnosol was purchased from Shanghai Standard Technology Co., Ltd.

[0112] Carnosol derivatives were provided by Sichuan University of Science & Engineering.

[0113] FBS (fetal bovine serum) was purchased from BI.

[0114] Horse serum was purchased from Gibco. High-glucose DMEM medium was purchased from Hyclone.

[0115] RPMI-1640 medium was purchased from Hyclone. Phenol red-free high-glucose DMEM medium was purchased from Hyclone.

[0116] P / S double antibody (penicillin-streptomycin mixture) was purchased from Hyclone. Dexamethasone was purchased from Sigma–Aldrich.

[0117] IBMX (3-isobutyl-1-methylxanthine), a broad-spectrum phosphodiesterase inhibitor, was purchased from Sigma–Aldrich.

[0118] Human recombinant insulin was purchased from Shanghai Jinmai Biotechnology.

[0119] The following is a detailed description of the specific embodiments provided by the present invention in conjunction with the accompanying drawings.

[0120] Synthesis of the target compound

[0121] Example 1: Synthesis of oxazolylsalviol

[0122]

[0123] Step 1: Preparation of I-2

[0124] Add I-1 (10.0 g, 30.5 mmol) and 100 mL of dry 1,4-dioxene to a 250 mL round-bottom flask, stir at room temperature until completely dissolved, slowly add 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) (7.61 g, 33.5 mmol), stir at room temperature for 0.5 h, and then detect the completion of the reaction by TLC. Filter the reaction solution under reduced pressure, and wash the filter cake with 50 mL of dichloromethane. Add 20 g of silica gel for column chromatography to the filtrate, distill under reduced pressure until there is no solvent, grind the residue into powder, pour it into a Buchner funnel, wash it with 200 mL of dichloromethane:ethyl acetate (50:1), evaporate the filtrate to dryness under reduced pressure, add 30 mL of absolute ethanol to the residue, heat under reflux to completely dissolve the solid, then slowly cool and crystallize, and filter to obtain I-2 (7.7 g, yield 77.5%).

[0125] 1 1H-NMR (600 MHz, Acetone-d6) δ 6.77 (s, 1H), 5.43 (s, 1H), 3.30 (dq, J = 13.6, 6.8 Hz, 1H), 2.85 - 2.79 (m, 1H), 2.54 (td, J = 13.8, 4.4 Hz, 1H), 2.25 - 2.16 (m, 1H), 1.96 (dt, J = 13.7, 3.2 Hz, 1H), 1.85 (ddd, J = 13.8, 10.7, 1.5 Hz, 1H), 1.69 (dd, J = 10.7, 5.8 Hz, 1H), 1.57 (dq, J = 10.5, 3.7 Hz, 1H), 1.54 - 1.47 (m, 1H), 1.32 (td, J = 13.4, 3.2 Hz, 1H), 1.19 (dd, J = 6.8, 3.9 Hz, 6H), 0.88 (d, J = 3.5 Hz, 6H).

[0126] Step 2: Preparation of I-3-1

[0127] Add Ⅰ-2 (3.0 g, 9.1 mmol) to a 100 mL round-bottom flask. Under nitrogen protection, add 50 mL of dry 1,4-dioxene. Stir at room temperature until completely dissolved. Slowly add 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) (2.27 g, 10.01 mmol). Stir at room temperature for 0.5 h. Monitor the reaction completion by TLC. Filter the reaction mixture under reduced pressure. Wash the filter cake with 20 mL of 1,4-dioxene. Evaporate the solvent under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether:dichloromethane = 2:3) to obtain a green solid Ⅰ-3-1 (2.61 g, yield 87.0%). ESI-MS m / z: 329 [M+H] + 。

[0128] 1 H NMR (600 MHz, Chloroform-d) δ 6.70 (s, 1H), 5.22 (s, 1H), 2.96 (hept, J = 7.8, 7.3 Hz, 1H), 2.67 (d, J = 14.3 Hz, 1H), 2.33–2.19 (m, 2H), 1.99 (ddd, J = 14.1, 10.7, 1.4 Hz, 1H), 1.88 (qt, J = 13.9, 3.3 Hz, 1H), 1.65 (ddd, J = 16.7, 10.3, 4.7 Hz, 2H), 1.52 (d, J = 14.8 Hz, 1H), 1.22 (td, J = 13.6, 3.5 Hz, 1H), 1.12 (d, J = 6.9 Hz, 6H), 0.88 (d, J = 21.1 Hz, 6H). 13 C NMR (150 MHz, Chloroform-d) δ 179.19, 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.

[0129] Step 3: Preparation of Ⅰ-4

[0130] Preparation of Compounds I-4-1 to I-4-30: Under nitrogen protection, 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. Monitor the reaction completion by TLC. Cool to room temperature, remove acetic acid under reduced pressure, add saturated sodium bicarbonate solution (20 mL), and extract with ethyl acetate (20 mL × 3). Add anhydrous sodium sulfate to the extract for drying, filter, and evaporate the solvent under reduced pressure. Purify the residue by silica gel column chromatography to obtain the corresponding products I-4-1 to I-4-30.

[0131]

[0132] Compound I-4-1: White solid, HR-MS m / z: C 21 H 25 NO3, 362.1727 [M+Na] + (Calculated), 362.1729 [M+Na] + (Found), 1 1H NMR (600 MHz, CDCl3) δ 8.08 (s, 1H), 7.16 (s, 1H), 5.66 (dd, J = 4.2, 1.8 Hz, 1H), 3.61 (p, J = 7.2 Hz, 1H), 3.00 (d, J = 13.8 Hz, 1H), 2.43 (td, J = 13.8, 4.8 Hz, 1H), 2.32 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.10 (qt, J = 13.8, 3.6 Hz, 1H), 1.96 (ddd, J = 13.8, 10.8, 1.8 Hz, 1H), 1.76 (dt, J = 13.8, 3.6 Hz, 1H), 1.69 (dd, J = 10.2, 6.0 Hz, 1H), 1.61 (d, J = 15.0 Hz, 1H), 1.37 (dd, J = 6.6, 3.0 Hz, 6H), 1.34–1.28 (m, 1H), 1.26 (s, 2H), 0.97 (s, 3H), 0.88 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.98, 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.

[0133]

[0134] Compound I-4-2: White solid, HR-MS m / z: C 27 H 29 NO3, 438.2040 [M+Na] + (Calculated), 438.2043 [M+Na] + (Found). 1 H NMR (600 MHz, CDCl3) δ 8.29–8.18 (m, 2H), 7.54 (dd, J = 5.4 1.8 Hz, 3H), 7.13 (s, 1H), 5.65 (dd, J = 4.2, 1.8 Hz, 1H), 3.68 (p, J = 7.2 Hz, 1H), 3.12 (d, J = 13.8 Hz, 1H), 2.54 (td, J = 13.8, 4.8 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.15 (qt, J = 13.8, 3.6 Hz, 1H), 1.98 (ddd, J = 13.8, 10.2, 1.8 Hz, 1H), 1.83 (dt, J = 13.8, 3.6 Hz, 1H), 1.74 (dd, J = 10.2, 6.0 Hz, 1H), 1.67–1.60 (m, 2H), 1.39 (dd, J = 6.6, 3.6 Hz, 6H), 1.34 (dd, J = 13.6, 3.6 Hz, 1H), 0.98 (s, 3H), 0.89 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 175.16, 162.43, 145.70, 140.16, 136.02, 131.51, 128.89, 127.68, 127.14, 120.90, 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.

[0135]

[0136] Compound I-4-3: White solid, ESI-MS m / z: 450 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 8.22 (t, J = 1.8 Hz, 1H), 8.12 (dt, J = 7.2, 1.2 Hz, 1H), 7.51 (dt, J = 7.8, 1.2 Hz, 1H), 7.47 (t, J = 7.8 Hz, 1H), 7.14 (s, 1H), 5.65 (dd, J = 4.2, 1.8 Hz, 1H), 3.66 (hept, J = 6.6 Hz, 1H), 3.11 (d, J = 13.8 Hz, 1H), 2.52 (td, J = 13.2, 4.8 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.15 (qt, J = 13.8, 3.6 Hz, 1H), 1.98 (ddd, J = 13.8, 10.2, 1.8 Hz, 1H), 1.84 (dq, J = 10.2, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.64 (d, J = 14.4 Hz, 1H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.02, 161.01, 145.76, 141.34, 140.46, 136.49, 135.02, 131.51, 130.24, 128.84, 127.55, 125.72, 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.

[0137]

[0138] Compound I-4-4: White solid, ESI-MS m / z: 450 [M + H] + , 11H NMR (600 MHz, CDCl3) δ 8.16 (dd, J = 7.2, 1.8 Hz, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.48–7.40 (m, 2H), 7.16 (s, 1H), 5.67 (d, J = 3.0 Hz, 1H), 3.67 (hept, J = 6.6 Hz, 1H), 3.08 (d, J = 13.8 Hz, 1H), 2.53 (td, J = 13.8, 4.2 Hz, 1H), 2.40–2.28 (m, 1H), 2.11 (ddd, J = 16.8, 8.4, 3.0 Hz, 1H), 2.03–1.94 (m, 1H), 1.78 (dt, J = 13.8, 3.0 Hz, 1H), 1.76–1.72 (m, 1H), 1.61 (d, J = 13.2 Hz, 1H), 1.40 (dd, J = 7.2, 3.6 Hz, 6H), 1.32 (td, J = 13.2, 3.6 Hz, 1H), 0.98 (s, 3H), 0.88 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.13, 160.56, 145.66, 140.97, 140.49, 136.48, 132.00, 131.37, 126.98, 126.28, 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.

[0139]

[0140] 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.

[0141]

[0142] 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). 1313C NMR (150 MHz, CDCl3) δ 175.06, 161.51, 145.70, 141.44, 140.31, 132.21, 129.07, 126.20, 126.06, 120.98, 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.

[0143]

[0144] Compound I-4-7: White solid, ESI-MS m / z: 434 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.24 (dd, J = 9.0, 5.4 Hz, 2H), 7.22 (t, J = 8.4 Hz, 2H), 7.13 (s, 1H), 5.65 (dd, J = 3.6, 1.2 Hz, 1H), 3.66 (hept, J = 6.6 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.52 (td, J = 13.2, 4.2 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.15 (qt, J = 13.8, 3.6 Hz, 1H), 2.03–1.93 (m, 1H), 1.82 (dt, J = 13.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.63 (d, J = 13.2 Hz, 1H), 1.39 (dd, J = 7.2, 3.0 Hz, 6H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.11, 165.64, 163.97, 161.54, 145.71, 141.49, 140.14, 136.07, 129.92, 123.47, 120.91, 116.25, 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.

[0145]

[0146] Compound I-4-8: White solid, ESI-MS m / z: 432 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 11.39 (s, 1H), 8.04–7.95 (m, 1H), 7.52–7.38 (m, 1H), 7.16 (s, 1H), 7.11 (d, J = 7.8 Hz, 1H), 7.03 (t, J = 7.2 Hz, 1H), 5.66 (dd, J = 3.6, 1.2 Hz, 1H), 3.56 (hept, J = 7.2 Hz, 1H), 3.19–3.06 (m, 1H), 2.62–2.48 (m, 1H), 2.34 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.16 (qt, J = 13.8, 3.0 Hz, 1H), 1.99 (ddd, J = 13.8, 10.8, 1.2 Hz, 1H), 1.84 (dq, J = 10.2, 3.6 Hz, 1H), 1.74 (dd, J = 10.2, 6.0 Hz, 1H), 1.65 (s, 1H), 1.42–1.33 (m, 6H), 0.99 (s, 3H), 0.90 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.85, 162.37, 158.65, 144.13, 139.20, 139.16, 136.55, 133.76, 127.09, 121.23, 119.73, 117.40, 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.

[0147]

[0148] 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.

[0149]

[0150] Compound I-4-10: White solid, ESI-MS m / z: 477 [M + H] + , 11H NMR (600 MHz, CDCl3) δ 7.75 (dd, J = 8.4, 1.8 Hz, 1H), 7.72 (d, J = 1.8 Hz, 1H), 7.11 (s, 1H), 7.04 (d, J = 8.4 Hz, 1H), 6.21 (s, 1H), 5.65 (dd, J = 3.6, 1.2 Hz, 1H), 3.99 (s, 3H), 3.69 (hept, J = 6.6 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.52 (td, J = 13.8, 4.2 Hz, 1H), 2.33 (ddd, J = 13.8, 6.0, 4.0 Hz, 1H), 2.14 (qt, J = 13.8, 3.0 Hz, 1H), 2.05–1.95 (m, 1H), 1.87–1.78 (m, 1H), 1.77–1.70 (m, 2H), 1.63 (d, J = 13.2 Hz, 1H), 1.43–1.31 (m, 7H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.34, 162.63, 149.03, 146.84, 145.56, 141.64, 139.67, 135.56, 121.86, 119.28, 115.82, 114.79, 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.

[0151]

[0152] Compound I-4-11: White solid, ESI-MS m / z: 446 [M + H] + , 11H NMR (600 MHz, CDCl3) δ 8.18 (d, J = 9.0 Hz, 2H), 7.10 (s, 1H), 7.03 (d, J = 9.0 Hz, 2H), 5.64 (dd, J = 3.6, 1.2 Hz, 1H), 3.90 (s, 3H), 3.66 (hept, J = 6.6 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.53 (td, J = 13.8, 4.2 Hz, 1H), 2.32 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.14 (qt, J = 13.2, 3.0 Hz, 1H), 2.01–1.94 (m, 1H), 1.82 (dt, J = 13.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.69 (d, J = 3.6 Hz, 1H), 1.63 (d, J = 13.2 Hz, 1H), 1.38 (dd, J = 6.6, 3.6 Hz, 6H), 1.34 (dd, J = 13.8, 3.6 Hz, 1H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.27, 162.56, 162.31, 145.56, 141.73, 139.67, 135.49, 129.45, 120.67, 119.68, 115.83, 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.

[0153]

[0154] Compound I-4-12: Yellow solid, ESI-MS m / z: 505 [M-H] - , 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, 40.86, 34.34, 31.68, 29.64, 29.20, 28.33, 23.13, 19.44, 18.70.

[0155]

[0156] 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), 2.03–1.95 (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). 1313C NMR (150 MHz, CDCl3) δ 174.99, 160.87, 145.84, 141.34, 140.70, 136.76, 133.08, 132.86, 130.40, 127.93, 125.92, 124.66, 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.

[0157]

[0158] Compound I-4-14: White solid, ESI-MS m / z: 474 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.32 (d, J = 8.4 Hz, 2H), 8.20 (d, J = 8.4 Hz, 2H), 7.15 (s, 1H), 5.69–5.64 (m, 1H), 3.98 (s, 3H), 3.68 (h, J = 6.6 Hz, 1H), 3.12 (d, J = 13.2 Hz, 1H), 2.54 (td, J = 13.8, 4.2 Hz, 1H), 2.34 (dt, J = 13.8, 4.2 Hz, 1H), 2.16 (q, J = 13.8 Hz, 1H), 2.03–1.96 (m, 1H), 1.85 (d, J = 13.8 Hz, 1H), 1.74 (dd, J = 10.2, 6.0 Hz, 1H), 1.64 (d, J = 13.2 Hz, 1H), 1.40 (dd, J = 6.6, 3.0 Hz, 6H), 1.38–1.33 (m, 1H), 0.99 (s, 3H), 0.90 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.02, 166.36, 161.33, 145.83, 141.45, 140.62, 136.67, 132.51, 130.99, 130.10, 127.54, 121.10, 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.

[0159]

[0160] Compound I-4-15: White solid, ESI-MS m / z: 458 [M-H] - ,1 1H NMR (600 MHz, DMSO-d6) δ 8.31 (d, J = 8.4 Hz, 2H), 8.16 (d, J = 8.4 Hz, 2H), 7.42 (s, 1H), 5.88 (d, J = 2.4 Hz, 1H), 3.60 (hept, J = 7.2 Hz, 1H), 2.92 (d, J = 13.8 Hz, 1H), 2.56 (dd, J = 13.8, 4.2 Hz, 1H), 2.28–2.19 (m, 1H), 2.00–1.88 (m, 2H), 1.75 (dd, J = 10.2, 6.0 Hz, 2H), 1.53 (d, J = 12.6 Hz, 1H), 1.37 (d, J = 6.6 Hz, 6H), 0.88 (s, 3H), 0.84 (s, 3H). 13 13C NMR (150 MHz, DMSO-d6) δ 174.87, 167.07, 161.43, 145.62, 140.99, 140.01, 137.44, 133.93, 130.60, 127.93, 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.

[0161]

[0162] Compound I-4-16: Yellow solid, ESI-MS m / z: 494 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.45 (d, J = 8.4 Hz, 2H), 8.12 (d, J = 8.4 Hz, 2H), 7.18 (s, 1H), 5.73–5.61 (m, 1H), 3.68 (hept, J = 6.6 Hz, 1H), 3.14 (s, 3H), 2.53 (td, J = 13.2, 4.2 Hz, 1H), 2.41–2.30 (m, 1H), 2.16 (q, 13.8 Hz, 1H), 2.05–1.93 (m, 1H), 1.85 (d, J = 10.2 Hz, 1H), 1.81–1.69 (m, 2H), 1.65 (d, J = 13.8 Hz, 1H), 1.40 (dd, J = 6.6, 2.4 Hz, 6H), 1.38–1.32 (m, 1H), 0.99 (s, 3H), 0.90 (s, 3H). 1313C NMR (150 MHz, CDCl3) δ 174.96, 160.31, 145.93, 142.67, 141.29, 140.95, 137.13, 132.01, 128.41, 128.05, 121.24, 117.93, 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.

[0163]

[0164] Compound I-4-17: White solid, ESI-MS m / z: 473 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.09 (s, 1H), 7.87 (d, J = 8.4 Hz, 2H), 7.51 (d, J = 8.4 Hz, 2H), 7.13 (s, 1H), 5.72–5.66 (m, 1H), 3.68 (hept, J = 7.2 Hz, 1H), 3.00 (d, J = 13.8 Hz, 1H), 2.46 (td, J = 13.2, 4.2 Hz, 1H), 2.37–2.31 (m, 1H), 2.24 (s, 3H), 2.12 (q, J = 13.8 Hz, 1H), 2.03–1.96 (m, 1H), 1.84 (d, J = 13.8 Hz, 1H), 1.74 (dd, J = 10.2, 6.0 Hz, 1H), 1.63 (d, J = 13.2 Hz, 1H), 1.49 (t, J = 7.8 Hz, 1H), 1.42 (dd, J = 20.4, 6.6 Hz, 6H), 1.38–1.32 (m, 2H), 0.99 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 176.33, 168.90, 161.99, 145.45, 141.73, 141.06, 140.05, 135.57, 128.30, 122.11, 120.31, 119.22, 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.

[0165]

[0166] 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.

[0167]

[0168] 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 = 7.8 Hz, 1H), 7.18 (s, 1H), 5.67 (d, J = 2.4 Hz, 1H), 3.68 (p, J = 6.6 Hz, 1H), 3.13 (d, J = 13.2 Hz, 1H), 2.53 (td, J = 13.2, 4.2 Hz, 1H), 2.40–2.28 (m, 1H), 2.23–2.06 (m, 1H), 2.05–1.94 (m, 1H), 1.86 (d, J = 14.4 Hz, 1H), 1.75 (dd, J = 10.2, 6.0 Hz, 1H), 1.64 (s, 1H), 1.45–1.38 (m, 7H), 0.99 (s, 3H), 0.91 (s, 3H).13 13C NMR (150 MHz, CDCl3) δ 174.89, 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.

[0169]

[0170] Compound I-4-20: White solid, ESI-MS m / z: 461 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 8.44–8.36 (m, 4H), 7.18 (s, 1H), 5.68 (dd, J = 3.6, 1.2 Hz, 1H), 3.67 (hept, J = 7.2 Hz, 1H), 3.13 (d, J = 13.8 Hz, 1H), 2.53 (td, J = 13.2, 4.2 Hz, 1H), 2.36 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.16 (qt, J = 13.8, 3.6 Hz, 1H), 2.06–1.94 (m, 1H), 1.85 (dq, J = 13.8, 3.6 Hz, 1H), 1.74 (dd, J = 10.2, 6.0 Hz, 1H), 1.65 (d, J = 11.4 Hz, 1H), 1.44–1.39 (m, 7H), 0.99 (s, 3H), 0.91 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 174.85, 160.02, 149.37, 146.00, 141.33, 141.06, 137.30, 132.70, 128.42, 124.20, 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.

[0171]

[0172] 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, 77.81, 47.11, 45.42, 40.87, 34.31, 31.67, 29.67, 28.23, 23.25, 23.09, 19.42, 18.67.

[0173]

[0174] Compound I-4-22: ESI-MS m / z: 478 [M+H] + , 11H 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.

[0175]

[0176] 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).13 13C NMR (150 MHz, CDCl3) δ 175.10, 156.86, 155.09, 144.89, 141.15, 140.81, 140.03, 135.98, 120.70, 116.01, 108.70, 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.

[0177]

[0178] Compound I-4-24: Yellow solid, ESI-MS m / z: 422 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 7.90 (d, J = 4.8 Hz, 1H), 7.55 (d, J = 6.0 Hz, 1H), 7.22–7.16 (m, 1H), 7.12 (s, 1H), 5.64 (dd, J = 3.6, 1.2 Hz, 1H), 3.66 (hept, J = 7.2 Hz, 1H), 3.07 (d, J = 13.8 Hz, 1H), 2.49 (td, J = 13.8, 4.2 Hz, 1H), 2.32 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.13 (qt, J = 13.8, 3.6 Hz, 1H), 2.04–1.90 (m, 1H), 1.80 (dq, J = 10.8, 3.6 Hz, 1H), 1.73 (dd, J = 10.2, 6.0 Hz, 1H), 1.62 (dd, J = 13.2, 1.2 Hz, 1H), 1.40–1.31 (m, 7H), 0.98 (s, 3H), 0.89 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.12, 158.40, 145.31, 141.48, 139.95, 136.03, 130.22, 130.01, 129.58, 128.20, 120.74, 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.

[0179]

[0180] 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.

[0181]

[0182] 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). 1313C NMR (150 MHz, CDCl3) δ 175.03, 161.02, 150.62, 145.93, 145.89, 141.27, 141.23, 137.12, 137.04, 125.57, 123.59, 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.

[0183]

[0184] Compound I-4-27: Yellow oil, ESI-MS m / z: 438 [M+H] + , 1 1H NMR (600 MHz, CDCl3) δ 7.08 (s, 1H), 5.63 (dd, J = 3.6, 1.2 Hz, 1H), 3.58 (hept, J = 7.2 Hz, 1H), 2.97 (d, J = 14.4 Hz, 1H), 2.93 (t, J = 7.8 Hz, 2H), 2.39 (td, J = 13.8, 4.8 Hz, 1H), 2.30 (ddd, J = 13.8, 6.0, 4.0 Hz, 1H), 2.09 (qt, J = 13.8, 3.0 Hz, 1H), 1.98–1.91 (m, 1H), 1.86 (p, J = 7.8 Hz, 2H), 1.69 (dd, J = 10.8, 6.0 Hz, 1H), 1.60 (d, J = 14.4 Hz, 1H), 1.46–1.41 (m, 2H), 1.38–1.23 (m, 20H), 0.96 (s, 3H), 0.90–0.87 (m, 6H). 13 13C NMR (150 MHz, CDCl3) δ 175.26, 166.83, 145.72, 140.70, 139.39, 135.33, 120.66, 116.17, 115.42, 77.87, 47.06, 45.41, 40.92, 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.

[0185]

[0186] Compound I-4-28: Yellow oil, ESI-MS m / z: 396 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 7.08 (s, 1H), 5.63 (dd, J = 4.2, 1.8 Hz, 1H), 3.58 (hept, J = 7.2 Hz, 1H), 2.99–2.92 (m, 3H), 2.78–2.73 (m, 1H), 2.54 (dt, J = 27.0, 7.8 Hz, 1H), 2.39 (td, J = 13.8, 4.8 Hz, 1H), 2.30 (ddd, J = 13.8, 6.0, 4.2 Hz, 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 13C NMR (150 MHz, CDCl3) δ 175.29, 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.

[0187]

[0188] 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.

[0189] Compound I-4-30: White solid, ESI-MS m / z: 481 [M+Na] + , 1 1H 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). 13CNMR(150MHz,CDCl3)δ175.84,163.84,152.40,149.31,138.97,134.90,129.83,129.17,128.09,115.75,114.40,111.52,77.77,48.44,45.47,40.98,40.15,34.36,31.79,29.88,29.28,28.65,22.60,22.54,19.54,18.78.

[0190]

[0191] 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 C NMR(150MHz,Chloroform-d)δ175.16,145.70,140.16,136.02,131.51,128.89,127.68,127.14,116.02,77.90,47.16,45.49,40.94,34.33,31.69,29.74,29.34,28.33,23.19,23.07,19.45,18.76. Example 2: Synthesis of benzene ring-substituted salvigenin derivatives

[0192]

[0193] Step 1: Preparation of Ⅰ-2

[0194] Add Ⅰ-1 (10.0 g, 30.5 mmol) and 100 mL of dry 1,4-dioxene to a 250 mL round-bottom flask. Stir at room temperature until completely dissolved. Slowly add 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) (7.61 g, 33.5 mmol). After stirring at room temperature for 0.5 h, monitor the reaction completion by TLC. Filter the reaction mixture under reduced pressure, and wash the filter cake with 50 mL of dichloromethane. Add 20 g of silica gel for column chromatography to the filtrate, and distill under reduced pressure until the solvent is removed. Grind the residue into a powder and pour it into a Buchner funnel. Wash it with 200 mL of dichloromethane:ethyl acetate (50:1). Evaporate the filtrate under reduced pressure to dryness. Add 30 mL of absolute ethanol to the residue and heat under reflux to completely dissolve the solid. Then slowly cool to crystallize. Filter to obtain Ⅰ-2 (7.7 g, yield 77.5%).

[0195] 1 H-NMR (600 MHz, Acetone-d6) δ 6.77 (s, 1H), 5.43 (s, 1H), 3.30 (dq, J = 13.6, 6.8 Hz, 1H), 2.85 - 2.79 (m, 1H), 2.54 (td, J = 13.8, 4.4 Hz, 1H), 2.25 - 2.16 (m, 1H), 1.96 (dt, J = 13.7, 3.2 Hz, 1H), 1.85 (ddd, J = 13.8, 10.7, 1.5 Hz, 1H), 1.69 (dd, J = 10.7, 5.8 Hz, 1H), 1.57 (dq, J = 10.5, 3.7 Hz, 1H), 1.54 - 1.47 (m, 1H), 1.32 (td, J = 13.4, 3.2 Hz, 1H), 1.19 (dd, J = 6.8, 3.9 Hz, 6H), 0.88 (d, J = 3.5 Hz, 6H).

[0196] Step 2: Preparation of Ⅰ-3-2

[0197]

[0198] Under nitrogen protection, compound I-2 (800 mg, 2.42 mmol) and dry dichloromethane (30 mL) were added to a 100 mL round-bottom flask, stirred until completely dissolved, then pyridine (780 μL, 764 mg, 9.68 mmol) was added. The temperature was cooled to 0 °C, stirred for 10 min, and then trifluoromethanesulfonic anhydride (900 μL, 1509 mg, 5.35 mmol) was slowly added dropwise. Stirred for 10 min, warmed to room temperature, and continued to react for 2 h. The reaction was monitored by TLC until completion. Dilute hydrochloric acid (30 mL, 0.1 M) was slowly added, and the layers were separated. The organic phase was washed with saturated sodium bicarbonate solution (30 mL). Anhydrous sodium sulfate was added to the organic phase for drying, and the solvent was evaporated under reduced pressure after filtration. 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), with a yield of 62.6%. ESI-MS m / z: 595 [M+Na] + , 1 H NMR (600 MHz, DMSO-d6) δ 7.86 (s, 1H), 5.83 (s, 1H), 3.10 (hept, J = 6.6 Hz, 1H), 2.74 (s, 1H), 2.22–2.13 (m, 1H), 1.98 (t, J = 10.2 Hz, 1H), 1.94–1.84 (m, 1H), 1.75 (qt, J = 13.8, 3.0 Hz, 1H), 1.63–1.52 (m, 2H), 1.40 (d, J = 13.2 Hz, 1H), 1.23 (d, J = 6.6 Hz, 4H), 1.14 (d, J = 6.6 Hz, 3H), 0.77 (s, 6H). 13 C NMR (150 MHz, DMSO-d6) δ 172.52, 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.

[0199] Step 3: Preparation of I-5

[0200]

[0201] Synthesis of Compounds I-5-1 to I-5-5: Under nitrogen protection, add Compound 35 (200 mg, 0.33 mmol), the corresponding boric acid derivative (1.32 mmol), potassium carbonate (140 mg, 1.01 mmol), tetrakis(triphenylphosphine)palladium (20 mg, 0.017 mmol) and dry toluene (10 mL) into a 25 mL round-bottom flask, heat to 90 °C, react for 6 - 20 h, and monitor the completion of the reaction by TLC. Cool to room temperature, add saturated sodium bicarbonate solution (20 mL), stir for 5 minutes and then separate the layers. Extract the aqueous phase with ethyl acetate (20 mL × 2), combine the organic phases, add anhydrous sodium sulfate to dry the organic phase, filter, and then evaporate the solvent under reduced pressure from the filtrate. The residue is purified by silica gel column chromatography to obtain Compounds I-5-1 to I-5-5.

[0202]

[0203] Compound I-5-1: White solid, ESI-MS m / z: 375 [M+H] + 。 1 H NMR (600 MHz, Methanol-d4) δ 7.32–7.26 (m, 3H), 7.27–7.21 (m, 1H), 7.12 (dd, J = 8.4, 1.2 Hz, 2H), 6.92 (s, 1H), 5.57 (dd, J = 4.2, 1.2 Hz, 1H), 2.92 (hept, J = 6.6 Hz, 1H), 2.33 (d, J = 13.8 Hz, 1H), 2.19 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 1.95–1.87 (m, 1H), 1.84 (ddd, J = 13.8, 10.8, 1.8 Hz, 1H), 1.71 (td, J = 13.2, 4.8 Hz, 1H), 1.55 (ddt, J = 13.8, 7.2, 3.6 Hz, 1H), 1.43 (dt, J = 10.8, 4.8 Hz, 2H), 1.18–1.10 (m, 2H), 1.03 (dd, J = 6.6, 2.4 Hz, 6H), 0.82 (s, 3H), 0.77 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 176.07, 145.31, 141.76, 141.21, 137.70, 137.41, 129.22, 128.09, 126.98, 123.57, 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.

[0204]

[0205] Compound I-5-2: Yellow solid, ESI-MS m / z: 393 [M+H] + , 1 H NMR(600 MHz, CDCl3) δ 7.26 (s, 1H), 7.20 (dd, J = 8.4, 5.4 Hz, 2H), 7.10 (t, J = 8.4 Hz, 2H), 7.02 (s, 1H), 5.59 (d, J = 2.4 Hz, 1H), 2.98 (p, J = 6.6 Hz, 1H), 2.57 (d, J = 13.2 Hz, 1H), 2.31 (ddd, J = 13.8, 6.0, 4.2 Hz, 1H), 2.07 (qt, J = 13.8, 3.6 Hz, 1H), 1.99 (ddd, J = 13.8, 10.8, 1.8 Hz, 1H), 1.81 (td, J = 13.2, 4.2 Hz, 1H), 1.69 (dt, J = 13.8, 3.6 Hz, 1H), 1.62–1.52 (m, 2H), 1.22 (td, J = 13.2, 3.6 Hz, 1H), 1.13 (dd, J = 25.2, 6.6 Hz, 6H), 0.96 (s, 3H), 0.87 (s, 3H). 13 C NMR(150 MHz, CDCl3) δ 175.98, 162.84, 161.21, 145.44, 140.13, 137.81, 137.62, 130.76, 130.71, 123.61, 119.93, 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.

[0206]

[0207] Compound I-5-3: Yellow solid, ESI-MS m / z: 409 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 7.38 (d, J = 8.4 Hz, 2H), 7.26 (s, 1H), 7.18 (d, J = 8.4 Hz, 2H), 7.01 (s, 1H), 5.59 (d, J = 2.4 Hz, 1H), 2.97 (hept, J = 7.2 Hz, 1H), 2.57 (d, J = 13.2 Hz, 1H), 2.35–2.28 (m, 1H), 2.07 (qt, J = 13.8, 3.6 Hz, 1H), 1.98 (ddd, J = 13.8, 10.8, 1.8 Hz, 1H), 1.80 (td, J = 13.2, 4.8 Hz, 1H), 1.69 (ddd, J = 13.8, 6.6, 3.6 Hz, 1H), 1.25–1.18 (m, 2H), 1.15 (d, J = 6.6 Hz, 3H), 1.11 (d, J = 6.6 Hz, 3H), 0.96 (s, 3H), 0.87 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 175.93, 145.34, 140.12, 139.91, 137.90, 137.77, 133.11, 130.54, 128.31, 123.43, 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.

[0208]

[0209] 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.

[0210]

[0211] Compound I-5-5: ESI-MS m / z: 405 [M+H] + , 11H NMR (600 MHz, CDCl3) δ 7.24 (s, 1H), 7.17 (d, J = 8.4 Hz, 2H), 7.04 (s, 1H), 6.95 (d, J = 8.4 Hz, 2H), 5.58 (d, J = 2.4 Hz, 1H), 3.86 (s, 3H), 3.06 (hept, J = 6.6 Hz, 1H), 2.57 (d, J = 12.6 Hz, 1H), 2.35–2.25 (m, 1H), 2.07 (qt, J = 13.6, 3.6 Hz, 1H), 2.02–1.95 (m, 1H), 1.68 (dt, J = 13.8, 3.6 Hz, 1H), 1.62–1.54 (m, 2H), 1.31–1.19 (m, 3H), 1.15 (d, J = 6.6 Hz, 3H), 1.11 (d, J = 6.6 Hz, 3H), 0.95 (s, 3H), 0.87 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 176.11, 158.67, 145.52, 140.84, 137.65, 137.20, 134.08, 130.26, 123.77, 119.84, 113.52, 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.

[0212] Example 3: Synthesis of 7-20 lactam salvianol

[0213]

[0214] Step 1: Synthesis of compound II-1

[0215] Under nitrogen protection, add salvianolic 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) to a 150 mL round-bottom flask, stir to dissolve the solid, heat to 70 °C, reflux for 20 h, and monitor the reaction by TLC until completion. Cool to room temperature, add saturated sodium bicarbonate solution (50 mL), stir for 10 min, then separate the layers. Extract the aqueous phase with dichloromethane (50 mL × 2), combine the organic phases, add anhydrous sodium sulfate to dry, filter, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether - ethyl acetate, volume ratio 15:1) to obtain 5.54 g of white solid (II-1) with a yield of 98.9%. ESI-MS m / z: 371 [M-H] - , 11H NMR (600 MHz, CDCl3) δ 6.42 (s, 1H), 3.29 (d, J = 13.2 Hz, 1H), 2.93–2.87 (m, 1H), 2.85 (dd, J = 15.6, 4.2 Hz, 1H), 2.77 (ddd, J = 16.8, 12.0, 6.6 Hz, 1H), 2.35 (qd, J = 12.6, 6.0 Hz, 1H), 2.06 (ddd, J = 17.4, 13.8, 10.2 Hz, 1H), 1.81 (dd, J = 13.2, 6.6 Hz, 1H), 1.61 (s, 3H), 1.54 (d, J = 13.2 Hz, 2H), 1.46 (d, J = 13.2 Hz, 1H), 1.33 (s, 3H), 1.27 (td, J = 13.2, 4.2 Hz, 1H), 1.20 (t, J = 6.6 Hz, 6H), 1.15 (dd, J = 13.8, 3.6 Hz, 1H), 0.95 (s, 3H), 0.83 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 180.97, 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.

[0216] Step 2: Synthesis of Compound Ⅱ-2

[0217] Under nitrogen protection, add Compound Ⅱ-1 (5.5 g, 14.8 mmol), potassium carbonate (3.05 g, 22.1 mmol) and acetone (60 mL) to a 150 mL round-bottom flask, stir until evenly suspended, then dropwise add methyl iodide (3.15 g, 22.1 mmol), heat to 65 °C, reflux for 10 h, and monitor the reaction by TLC until completion. Cool to room temperature, filter the reaction mixture under reduced pressure to remove potassium carbonate, wash the filter cake with dichloromethane (20 mL), evaporate the filtrate under reduced pressure, add saturated ammonium chloride solution (50 mL), then extract with dichloromethane (50 mL × 3). Add anhydrous sodium sulfate to the extract for drying, filter, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether - ethyl acetate, volume ratio 30:1) to obtain 5.5 g of a colorless oil (Ⅱ-2) with a yield of 96.5%. 11H NMR (600 MHz, CDCl3) δ 6.43 (s, 1H), 3.59 (s, 3H), 3.36 (d, J = 13.2 Hz, 1H), 2.95–2.85 (m, 2H), 2.79 (ddd, J = 17.4, 12.0, 6.0 Hz, 1H), 2.38 (qd, J = 12.6, 6.0 Hz, 1H), 2.05 (qt, J = 13.8, 4.2 Hz, 1H), 1.82 (dd, J = 13.2, 6.6 Hz, 1H), 1.65 (s, 3H), 1.58–1.55 (m, 1H), 1.54 (s, 3H), 1.46 (d, J = 13.2 Hz, 1H), 1.33–1.24 (m, 1H), 1.21 (dd, J = 7.2, 3.6 Hz, 6H), 0.96 (s, 3H), 0.76 (s, 3H).

[0218] Step 3: Synthesis of Compound II-3

[0219] Compound II-2 (5.5 g, 14.7 mmol) was added to acetic acid (30 mL), and the mixture was stirred until dissolved. Chromium trioxide (3.2 g, 32.5 mmol) was added, and the reaction was carried out at room temperature for 8 h. The reaction was monitored by TLC until completion. The acetic acid was evaporated under reduced pressure. Ethyl acetate (50 mL) and saturated sodium bicarbonate solution (50 mL) were added, and the mixture was stirred for 10 min. Then, liquid separation was carried out. The aqueous phase was extracted with ethyl acetate (50 mL × 2). 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, volume ratio 30:1) to obtain 3.8 g of a colorless oil (II-3) with a yield of 66.7%. 1 1H NMR (600 MHz, CDCl3) δ 7.63 (s, 1H), 3.61 (s, 3H), 3.51 (d, J = 13.8 Hz, 1H), 3.19 (dd, J = 18.0, 14.4 Hz, 1H), 2.99 (hept, J = 6.6 Hz, 1H), 2.65 (dd, J = 18.0, 4.8 Hz, 1H), 2.12 (dd, J = 14.4, 4.8 Hz, 1H), 1.99 (qt, J = 13.8, 3.6 Hz, 1H), 1.66 (d, J = 23.4 Hz, 6H), 1.50 (d, J = 13.2 Hz, 1H), 1.34 (m, 2H), 1.24 (dd, J = 10.8, 7.2 Hz, 6H), 0.94 (s, 3H), 0.82 (s, 3H).

[0220] Step 4: Synthesis of Compound II-4

[0221]

[0222] Synthesis of Compound Ⅱ-4-1: Under nitrogen protection, add 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) into a 100 mL round-bottom flask, stir to dissolve, heat to 85 °C, reflux for 12 h, monitor by TLC until Compound Ⅱ-3 disappears, cool to room temperature, evaporate ethanol under reduced pressure, add saturated sodium bicarbonate solution (50 mL), extract with ethyl acetate (50 mL×3), add anhydrous sodium sulfate to the extract for drying, filter and evaporate the solvent under reduced pressure. Under nitrogen protection, add the residue into a 250 mL flask, then add ammonium acetate (4.0 g, 52.0 mmol), sodium cyanoborohydride (1.6 g, 26.0 mmol) and methanol (80 mL), cool to 0 °C, stir for 10 min, then add titanium trichloride solution (mass fraction 15%, 6.4 mL, containing 960 mg, 6.2 mmol of titanium trichloride), react at 0 °C for 1 h and then restore to room temperature, continue to react for 6 h, monitor the reaction by TLC until it is completed. Evaporate the solvent under reduced pressure, add saturated sodium bicarbonate solution (50 mL), extract with ethyl acetate (50 mL×3), add anhydrous sodium sulfate to the extract for drying, filter, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (dichloromethane - methanol, volume ratio 30:1) to obtain 1.81 g of a colorless oil (Ⅱ-4-1), with a yield of 87.9%. ESI-MS m / z: 402 [M+H] + , 1 H NMR (600 MHz, CDCl3) δ 6.80 (s, 1H), 4.64 (dd, J = 9.6, 5.4 Hz, 1H), 3.73 (s, 3H), 3.43 (d, J = 13.8 Hz, 1H), 2.96 (hept, J = 7.2 Hz, 1H), 2.56 (ddd, J = 14.4, 9.6, 4.8 Hz, 1H), 2.14 (td, J = 13.8, 6.0 Hz, 1H), 1.93 (m, 1H), 1.70–1.58 (m, 8H), 1.48 (d, J = 13.2 Hz, 1H), 1.23 (dd, J = 12.0, 7.2 Hz, 6H), 0.96 (s, 3H), 0.77 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 176.77, 145.44, 144.62, 130.30, 124.96, 122.44, 120.90, 117.62, 52.89, 51.10, 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.

[0223]

[0224] Synthesis of Compounds Ⅱ-4-2 to Ⅱ-4-9: Under nitrogen protection, add Compound Ⅱ-3 (200 mg, 0.52 mmol), the corresponding amine (1.0 mmol), and 1,4-dioxane (8 mL) into a 50 mL round-bottom flask. Stir until dissolved, then add titanium(IV) isopropoxide (284 mg, 1.0 mmol). Heat to 110 °C and reflux for about 12 h. Monitor the disappearance of Compound Ⅱ-3 by TLC. Cool to room temperature, add absolute ethanol (10 mL) to the reaction flask, and then slowly add sodium borohydride (190 mg, 5.2 mmol). React at room temperature for about 5 h and detect the completion of the reaction by TLC. Add water (1 mL) to quench the reaction. At this time, a large amount of white solid appears. Filter the reaction solution, wash the filter cake with dichloromethane (20 mL), collect the filtrate, evaporate the solvent under reduced pressure. Add saturated sodium bicarbonate solution (20 mL) to the residue, extract with ethyl acetate (20 mL × 3). Add anhydrous sodium sulfate to dry the extract, filter and evaporate the solvent. Purify the residue by silica gel column chromatography.

[0225]

[0226] Compound Ⅱ-4-2: Colorless oil, ESI-MS m / z: 492 [M+H] + , 1 H NMR (600 MHz, CDCl3) δ 7.41 (d, J = 7.2 Hz, 2H), 7.33 (t, J = 7.8 Hz, 2H), 7.24 (t, J = 7.2 Hz, 1H), 7.11 (s, 1H), 3.95 (dd, J = 10.8, 6.7 Hz, 1H), 3.80 (q, J = 13.2 Hz, 2H), 3.61 (s, 3H), 3.38 (d, J = 13.2 Hz, 1H), 2.96 (hept, J = 7.2 Hz, 1H), 2.57–2.40 (m, 1H), 2.20–2.12 (m, 1H), 2.06 (qt, J = 13.8, 3.6 Hz, 1H), 1.64 (s, 3H), 1.62–1.53 (m, 5H), 1.47 (d, J = 13.2 Hz, 1H), 1.24 (dd, J = 13.8, 6.6 Hz, 6H), 1.15 (td, J = 13.2, 4.2 Hz, 1H), 0.97 (s, 3H), 0.80 (s, 3H). 1313C NMR (150 MHz, CDCl3) δ 175.09, 144.32, 143.23, 141.35, 132.30, 128.31, 126.74, 122.25, 118.06, 116.53, 57.10, 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.

[0227]

[0228] Compound Ⅱ-4-3: Colorless oil, 1 1H NMR (600 MHz, CDCl3) δ 7.35 (d, J = 8.4 Hz, 2H), 7.29 (d, J = 8.4 Hz, 2H), 7.09 (s, 1H), 3.93 (dd, J = 10.8, 6.6 Hz, 1H), 3.77 (q, J = 13.2 Hz, 2H), 3.62 (s, 3H), 3.38 (d, J = 13.2 Hz, 1H), 2.95 (hept, J = 7.2 Hz, 1H), 2.52–2.43 (m, 1H), 2.14 (dd, J = 12.6, 6.6 Hz, 1H), 2.05 (dddt, J = 17.4, 13.8, 8.4, 3.6 Hz, 1H), 1.64 (s, 3H), 1.57 (q, J = 6.0, 5.4 Hz, 5H), 1.47 (d, J = 13.2 Hz, 1H), 1.27–1.20 (m, 8H), 1.15 (td, J = 13.2, 4.2 Hz, 1H), 0.97 (s, 3H), 0.79 (s, 3H).

[0229]

[0230] Compound Ⅱ-4-4: Colorless oil, 11H NMR (600 MHz, CDCl3) δ 7.54–7.41 (m, 4H), 7.04 (s, 1H), 3.87 (dd, J = 10.2, 6.6 Hz, 1H), 3.79 (q, J = 13.2 Hz, 2H), 3.55 (s, 3H), 3.32 (d, J = 13.2 Hz, 1H), 2.89 (hept, J = 7.2 Hz, 1H), 2.48–2.36 (m, 1H), 2.07 (ddd, J = 12.6, 6.6, 1.8 Hz, 1H), 1.98 (qt, J = 13.8, 3.6 Hz, 1H), 1.57 (s, 3H), 1.52–1.46 (m, 5H), 1.40 (d, J = 13.1 Hz, 1H), 1.16 (t, J = 7.2 Hz, 6H), 1.08 (td, J = 13.2, 4.2 Hz, 1H), 0.90 (s, 3H), 0.72 (s, 3H).

[0231]

[0232] Compound II-4-5: Colorless oil, 1 1H NMR (600 MHz, CDCl3) δ 8.56–8.52 (d, J = 6.0 Hz, 2H), 7.37 (d, J = 6.0 Hz, 2H), 7.13 (s, 1H), 3.95 (dd, J = 10.2, 6.6 Hz, 1H), 3.88–3.78 (m, 2H), 3.63 (s, 3H), 3.39 (d, J = 13.2 Hz, 1H), 2.96 (hept, J = 7.2 Hz, 1H), 2.55–2.41 (m, 1H), 2.16–2.10 (m, 1H), 2.05 (tdd, J = 17.4, 7.8, 4.8 Hz, 1H), 1.64 (s, 4H), 1.58 (s, 4H), 1.50–1.45 (m, 1H), 1.23 (dd, J = 6.6, 1.2 Hz, 7H), 1.15 (dd, J = 12.0, 2.4 Hz, 1H), 0.97 (s, 3H), 0.79 (s, 3H).

[0233]

[0234] Compound II-4-6: Colorless oil, 11H NMR (600 MHz, CDCl3) δ 7.23–7.20 (m, 1H), 7.17 (s, 1H), 6.96–6.92 (m, 2H), 4.03 (d, J = 6.6 Hz, 1H), 4.01–3.91 (m, 2H), 3.61 (s, 3H), 3.38 (d, J = 13.2 Hz, 1H), 2.96 (hept, J = 7.2 Hz, 1H), 2.50–2.42 (m, 1H), 2.13 (ddd, J = 12.6, 6.6, 1.8 Hz, 1H), 2.05 (dddd, J = 17.4, 13.8, 9.0, 3.6 Hz, 1H), 1.64 (s, 3H), 1.60–1.53 (m, 5H), 1.50–1.44 (m, 1H), 1.25 (dd, J = 10.8, 7.2 Hz, 6H), 1.15 (td, J = 13.2, 4.2 Hz, 1H), 0.97 (s, 3H), 0.79 (s, 3H).

[0235]

[0236] Compound Ⅱ-4-7: White solid, 1 1H NMR (600 MHz, CDCl3) δ 7.52 (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, 4.8 Hz, 2H), 2.93 (p, J = 7.2 Hz, 1H), 2.90–2.76 (m, 2H), 2.45–2.36 (m, 1H), 2.20 (ddd, J = 12.6, 7.2, 2.4 Hz, 1H), 2.02 (qt, J = 13.8, 3.6 Hz, 1H), 1.63 (s, 3H), 1.58 (m, 5H), 1.47 (d, J = 13.2 Hz, 1H), 1.31–1.24 (m, 2H), 1.20 (d, J = 7.2 Hz, 6H), 1.15 (td, J = 13.2, 4.2 Hz, 1H), 0.96 (s, 3H), 0.76 (s, 3H).

[0237]

[0238] Compound Ⅱ-4-8: White solid, 11H NMR (600 MHz, CDCl3) δ 6.92 (s, 1H), 4.15 (s, 1H), 3.73 (s, 4H), 3.64 (s, 3H), 3.40 (d, J = 13.8 Hz, 1H), 2.92 (hept, J = 7.2 Hz, 1H), 2.75 (ddt, J = 24.0, 12.0, 5.4 Hz, 2H), 2.63 (dt, J = 12.0, 6.0 Hz, 1H), 2.56 (dt, J = 12.0, 5.4 Hz, 1H), 2.53–2.37 (m, 5H), 2.15 (s, 1H), 2.02 (qt, J = 13.8, 3.6 Hz, 1H), 1.63 (s, 3H), 1.59 (m, 4H), 1.47 (d, J = 13.2 Hz, 1H), 1.23 (t, J = 7.2 Hz, 6H), 1.15 (td, J = 13.2, 4.2 Hz, 1H), 0.97 (s, 3H), 0.77 (s, 3H).

[0239]

[0240] Compound Ⅱ-4-9: Yield 78.2%, yellow solid, 1 1H NMR (600 MHz, CDCl3) δ 6.93 (s, 1H), 3.78 (dd, J = 10.2, 6.6 Hz, 1H), 3.53 (s, 3H), 3.30 (d, J = 13.2 Hz, 1H), 3.12 (d, J = 11.4 Hz, 2H), 2.61 (t, J = 12.0 Hz, 2H), 2.47–2.40 (m, 2H), 2.30 (q, J = 12.0 Hz, 1H), 2.03–1.93 (m, 2H), 1.89 (s, 1H), 1.79 (d, J = 12.6 Hz, 2H), 1.56 (s, 3H), 1.50 (s, 3H), 1.39 (d, J = 12.6 Hz, 1H), 1.17–1.11 (m, 6H), 1.10–1.03 (m, 1H), 0.89 (s, 3H), 0.70 (s, 3H).

[0241] Step 5: Synthesis of Compound Ⅱ-5

[0242]

[0243] Synthesis of Compounds Ⅱ-5-1 to Ⅱ-5-9: Under nitrogen protection, corresponding Compounds Ⅱ-4-1 to Ⅱ-4-9 (about 200 mg, 0.50 mmol), sodium hydroxide (150 mg, 3.75 mmol), and absolute ethanol (10 mL) were added to a 25 mL round-bottom flask. The mixture was heated to 85 °C and refluxed for about 10 h. The reaction was monitored by TLC until completion. After cooling to room temperature, acetic acid (1 mL) was slowly added to neutralize sodium hydroxide. The solvent was evaporated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). Anhydrous sodium sulfate was added to the extract for drying. After filtration, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the corresponding product.

[0244]

[0245] Compound Ⅱ-5-1, colorless oil, ESI-MS m / z: 370 [M+H] + , 1 H NMR (600 MHz, CDCl3) δ 7.00 (s, 1H), 6.36 (s, 1H), 4.34 (ddd, J = 5.4, 3.6, 2.4 Hz, 1H), 2.87 (hept, J = 7.2 Hz, 1H), 2.66 (d, J = 12.6 Hz, 1H), 2.02–1.88 (m, 3H), 1.81–1.69 (m, 2H), 1.61 (s, 3H), 1.55 (dd, J = 9.6, 3.0 Hz, 1H), 1.50 (s, 3H), 1.44 (d, J = 13.2 Hz, 1H), 1.18 (t, J = 4.8 Hz, 1H), 1.11 (dd, J = 9.6, 7.2 Hz, 6H), 0.82 (s, 3H), 0.75 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 177.07, 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.

[0246]

[0247] Compound Ⅱ-5-2: colorless oil, 11H NMR (600 MHz, CDCl3) δ 7.19 (d, J = 7.8 Hz, 4H), 7.12 (d, J = 7.8 Hz, 2H), 6.18 (s, 1H), 4.59 (d, J = 15.0 Hz, 1H), 4.29 (d, J = 15.0 Hz, 1H), 4.17 (t, J = 2.4 Hz, 1H), 2.91–2.80 (m, 1H), 2.73 (d, J = 10.8 Hz, 1H), 2.01 (q, J = 12.0, 10.2 Hz, 2H), 1.70 (ddd, J = 12.6, 6.0, 3.6 Hz, 1H), 1.68–1.62 (m, 1H), 1.60 (s, 3H), 1.57 (dd, J = 10.2, 5.4 Hz, 2H), 1.54 (s, 3H), 1.43 (d, J = 13.2 Hz, 1H), 1.18 (s, 1H), 1.08 (t, J = 6.6 Hz, 6H), 0.75 (s, 3H), 0.70 (s, 3H).

[0248]

[0249] Compound II-4-3: White solid, 1 1H NMR (600 MHz, CDCl3) δ 7.22 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 8.4 Hz, 2H), 6.23 (s, 1H), 4.55 (d, J = 15.0 Hz, 1H), 4.40 (d, J = 15.0 Hz, 1H), 4.21 (t, J = 2.4 Hz, 1H), 2.92 (hept, J = 7.2 Hz, 1H), 2.79 (d, J = 10.8 Hz, 1H), 2.12–2.00 (m, 2H), 1.81–1.76 (m, 1H), 1.76–1.70 (m, 1H), 1.66 (s, 3H), 1.64 (dd, J = 10.2, 6.0 Hz, 2H), 1.61 (s, 3H), 1.50 (d, J = 13.2 Hz, 1H), 1.25 (t, J = 13.2 Hz, 1H), 1.15 (t, J = 6.6 Hz, 6H), 0.80 (s, 3H), 0.77 (s, 3H).

[0250]

[0251] Compound II-5-4: Colorless oil, 11H NMR (600 MHz, CDCl3) δ 7.51 (d, J = 7.8 Hz, 2H), 7.26 (d, J = 7.8 Hz, 2H), 6.23 (s, 1H), 4.63–4.51 (m, 2H), 4.25–4.17 (m, 1H), 2.92 (hept, J = 7.2 Hz, 1H), 2.80 (d, J = 11.2 Hz, 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 = 13.2 Hz, 1H), 1.26 (dd, J = 8.4, 4.2 Hz, 1H), 1.14 (dd, J = 11.8, 7.2 Hz, 6H), 0.81 (d, J = 14.4 Hz, 6H).

[0252]

[0253] Compound Ⅱ-5-5: Colorless oil, 1 1H NMR (600 MHz, CDCl3) δ 8.48 (d, J = 6.0 Hz, 2H), 7.05 (d, J = 6.0 Hz, 2H), 6.28 (s, 1H), 4.58–4.49 (m, 2H), 4.22–4.19 (m, 1H), 2.93 (hept, J = 7.2 Hz, 1H), 2.80 (d, J = 11.4 Hz, 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 = 13.8 Hz, 1H), 1.28–1.23 (m, 2H), 1.16 (dd, J = 6.6, 4.2 Hz, 6H), 0.83 (s, 3H), 0.81 (s, 3H).

[0254]

[0255] Compound Ⅱ-5-6: Colorless oil, 11H NMR (600 MHz, CDCl3) δ 7.21 (dd, J = 4.8, 1.2 Hz, 1H), 6.95 (d, J = 2.4 Hz, 1H), 6.93 (dd, J = 4.8, 3.6 Hz, 1H), 6.34 (s, 1H), 4.97 (d, J = 15.0 Hz, 1H), 4.39–4.34 (m, 2H), 2.93 (hept, J = 7.2 Hz, 1H), 2.78 (d, J = 7.8 Hz, 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 = 11.2 Hz, 1H), 1.25 (t, J = 13.2 Hz, 1H), 1.17 (t, J = 7.2 Hz, 6H), 0.81 (s, 3H), 0.77 (s, 3H).

[0256]

[0257] Compound Ⅱ-5-7: White solid, 1 1H NMR (600 MHz, CDCl3) δ 7.63 (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 = 7.2 Hz, 4H), 2.86 (m, 2H), 2.77 (m, 1H), 2.63 (d, J = 13.8 Hz, 1H), 2.01 (td, J = 13.8, 4.2 Hz, 1H), 1.92 (q, J = 13.2 Hz, 1H), 1.86–1.80 (m, 1H), 1.73 (t, J = 12.6 Hz, 1H), 1.59 (s, 3H), 1.56 (dd, J = 10.2, 6.0 Hz, 2H), 1.52 (s, 3H), 1.40 (d, J = 13.2 Hz, 2H), 1.32 (t, J = 7.2 Hz, 6H), 1.12 (dd, J = 6.6, 4.8 Hz, 6H), 0.71 (s, 3H), 0.64 (s, 3H).

[0258]

[0259] Compound Ⅱ-5-8: Yellow oil, 11H NMR (600 MHz, CDCl3) δ 6.44 (s, 1H), 4.38–4.33 (m, 1H), 3.69–3.62 (m, 4H), 3.59 (dt, J = 13.8, 6.0 Hz, 1H), 3.34 (m, 1H), 2.95 (hept, J = 7.2 Hz, 1H), 2.74 (d, J = 11.4 Hz, 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), 1.58 (s, 3H), 1.49 (d, J = 13.2 Hz, 1H), 1.19 (dd, J = 11.4, 6.6 Hz, 6H), 0.82 (d, J = 13.2 Hz, 6H).

[0260]

[0261] Compound Ⅱ-5-9: 77.3%, yield colorless oil, 1 1H NMR (600 MHz, CDCl3) δ 6.44 (s, 1H), 4.28–4.23 (m, 1H), 3.33 (dd, J = 13.8, 7.8 Hz, 1H), 3.08–2.98 (m, 3H), 2.95 (p, J = 7.2 Hz, 1H), 2.74 (d, J = 10.2 Hz, 1H), 2.54 (td, J = 12.0, 2.4 Hz, 1H), 2.44 (td, J = 12.0, 2.4 Hz, 1H), 2.09–2.00 (m, 2H), 1.96 (ddd, J = 12.6, 5.4, 3.6 Hz, 1H), 1.85–1.79 (m, 1H), 1.66 (s, 3H), 1.59 (s, 3H), 1.49 (d, J = 12.0 Hz, 1H), 1.40 (d, J = 13.8 Hz, 1H), 1.18 (dd, J = 14.4, 6.6 Hz, 7H), 1.13–1.05 (m, 2H), 0.81 (d, J = 2.4 Hz, 6H).

[0262] Step 6: Synthesis of Compound Ⅱ-6

[0263]

[0264] Synthesis of Compounds Ⅱ-6-1 to Ⅱ-6-8: The corresponding Compounds Ⅱ-5-1 to Ⅱ-5-8 (about 100 mg, 0.27 mmol) were added to dichloromethane (5 mL), stirred and dissolved. Then trifluoroacetic acid (5 mL) was slowly added, and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC until completion. The solvent was evaporated under reduced pressure. Saturated sodium bicarbonate solution (20 mL) was added to the residue, and the mixture was extracted with dichloromethane (20 mL × 3). Anhydrous sodium sulfate was added to the extract for drying. After filtration, dichloromethane was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the corresponding products Ⅱ-6-1 to Ⅱ-6-8.

[0265]

[0266] Compound Ⅱ-6-1: White solid, ESI-MS m / z: 328 [M-H] - , 1 H NMR (600 MHz, DMSO-d6) δ 8.17 (d, J = 5.4 Hz, 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 = 13.2 Hz, 1H), 2.32 (td, J = 13.8, 4.2 Hz, 1H), 1.92–1.77 (m, 2H), 1.63 (t, J = 10.8 Hz, 1H), 1.50 (dd, J = 10.2, 5.4 Hz, 1H), 1.45 (d, J = 13.2 Hz, 1H), 1.40 (d, J = 12.6 Hz, 1H), 1.21–1.15 (m, 1H), 1.10 (t, J = 6.6 Hz, 6H), 0.79 (d, J = 16.2 Hz, 6H). 13 C NMR (150 MHz, DMSO-d6) δ 177.04, 143.67, 141.65, 136.82, 133.43, 125.07, 110.07, 51.74, 50.01, 47.24, 41.60, 34.98, 32.11, 31.75, 29.24, 26.58, 23.42, 23.20, 19.94, 19.57.

[0267]

[0268] Compound Ⅱ-6-2: White solid, ESI-MS m / z: 418 [M-H] - , 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.

[0269]

[0270] Compound II-6-3: White solid, ESI-MS m / z: 453 [M-H] - , 1 1H NMR (600 MHz, CDCl3) δ 7.04 (d, J = 8.4 Hz, 2H), 6.84 (d, J = 8.4 Hz, 2H), 6.15 (s, 1H), 4.45 (d, J = 15.0 Hz, 1H), 4.27 (d, J = 15.0 Hz, 1H), 4.12–4.08 (m, 1H), 3.06 (d, J = 12.6 Hz, 1H), 2.98 (hept, J = 7.2 Hz, 1H), 2.33 (td, J = 13.2, 4.2 Hz, 1H), 1.99 (q, J = 13.8 Hz, 1H), 1.74 (dt, J = 12.0, 4.2 Hz, 1H), 1.68–1.63 (m, 1H), 1.62–1.58 (m, 2H), 1.43 (d, J = 12.6 Hz, 1H), 1.21–1.16 (m, 1H), 1.01 (d, J = 6.6 Hz, 3H), 0.96 (d, J = 6.6 Hz, 3H), 0.71 (d, J = 4.2 Hz, 6H). 1313C NMR (150 MHz, CDCl3) δ 175.68, 141.95, 140.83, 135.11, 134.41, 133.20, 132.50, 124.49, 111.32, 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.

[0271]

[0272] Compound Ⅱ-6-4: ESI-MS m / z: 486 [M-H] - , 1 1H NMR (600 MHz, CDCl3) δ 7.35 (d, J = 8.4 Hz, 2H), 7.05 (d, JⅡ-6-4 = 7.8 Hz, 2H), 6.16 (s, 1H), 4.59 (d, J = 15.0 Hz, 1H), 4.35 (d, J = 15.0 Hz, 1H), 4.13–4.09 (m, 1H), 3.03 (d, J = 12.0 Hz, 1H), 2.97 (hept, J = 7.2 Hz, 1H), 2.32 (td, J = 13.2, 4.2 Hz, 1H), 2.00 (qt, J = 13.8, 3.0 Hz, 1H), 1.79 (dt, J = 12.0, 4.1 Hz, 1H), 1.71–1.59 (m, 3H), 1.45 (d, J = 13.2 Hz, 1H), 1.02 (d, J = 6.6 Hz, 3H), 0.95 (d, J = 6.6 Hz, 3H), 0.73 (d, J = 6.0 Hz, 6H). 13 13C NMR (150 MHz, CDCl3) δ 174.61, 140.69, 139.75, 133.38, 131.38, 127.63, 124.25, 123.21, 110.25, 57.00, 49.57, 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.

[0273]

[0274] Compound Ⅱ-6-5: ESI-MS m / z: 419 [M-H] - , 11H NMR (600 MHz, CDCl3) δ 8.29 (d, J = 6.0 Hz, 2H), 6.92 (d, J = 6.0 Hz, 2H), 6.29 (s, 1H), 4.81 (d, J = 15.6 Hz, 1H), 4.27 (d, J = 15.6 Hz, 1H), 4.22–4.17 (m, 1H), 3.15 (hept, J = 6.6 Hz, 1H), 3.00 (d, J = 13.8 Hz, 1H), 2.43 (td, J = 13.2, 4.2 Hz, 1H), 2.01 (qt, J = 13.8, 3.0 Hz, 1H), 1.93 (ddd, J = 12.6, 5.4, 3.6 Hz, 1H), 1.83–1.77 (m, 1H), 1.72 (dd, J = 10.2, 5.4 Hz, 1H), 1.64 (dt, J = 13.8, 3.6 Hz, 1H), 1.50 (d, J = 13.2 Hz, 1H), 1.10 (d, J = 6.6 Hz, 3H), 1.02 (d, J = 6.6 Hz, 3H), 0.81 (d, J = 3.0 Hz, 6H). 13 13C NMR (150 MHz, CDCl3) δ 175.84, 148.89, 147.20, 142.05, 141.77, 134.64, 133.14, 124.15, 123.29, 110.87, 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.

[0275]

[0276] Compound Ⅱ-6-6: Yellow solid, ESI-MS m / z: 424 [M-H] - , 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.

[0277]

[0278] 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). 1313C NMR (150 MHz, Methanol-d4) δ 176.37, 143.03, 141.51, 135.22, 134.45, 134.07, 133.53, 124.10, 116.53, 109.83, 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.

[0279]

[0280] Compound Ⅱ-6-8, white solid, ESI-MS m / z: 441 [M-H] - , 1 1H NMR (600 MHz, CDCl3) δ 6.52 (s, 1H), 4.36–4.29 (m, 1H), 3.70–3.58 (m, 4H), 3.50 (dt, J=13.2, 6.0 Hz, 1H), 3.32 (dt, J=13.8, 7.2 Hz, 1H), 3.18 (hept, J=6.6 Hz, 1H), 3.03 (d, J=13.2 Hz, 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=13.2 Hz, 1H), 1.26 (td, J=13.2, 4.2 Hz, 1H), 1.17 (dd, J=15.6, 7.2 Hz, 6H), 0.81 (d, J=4.2 Hz, 6H). 13 13C NMR (150 MHz, CDCl3) δ 175.40, 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. Example 4: Synthesis of Salvianolic Acid B Derivatives Based on Compound Ⅱ-6-1

[0281]

[0282] Synthesis of Compound Ⅱ-7-1: Under nitrogen protection, add Compound Ⅱ-6-1 (100 mg, 0.30 mmol), potassium carbonate (126 mg, 0.90 mmol) and acetone (5 mL) into a 25 mL single-necked flask. Stir until evenly dispersed, then add methyl iodide (127 mg, 0.90 mmol) dropwise. Heat to 65 °C and reflux for 20 h. Monitor the reaction by TLC until completion. Cool to room temperature, filter the reaction mixture under reduced pressure to remove potassium carbonate, evaporate the filtrate under reduced pressure to dryness. Add saturated ammonium chloride solution (20 mL) to the residue, extract with ethyl acetate (20 mL × 3). Add anhydrous sodium sulfate to the extract for drying, filter, and evaporate the solvent under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether - ethyl acetate, volume ratio 7:2) to obtain 84 mg of yellow solid (Ⅱ-7-1), with a yield of 77.0%. ESI-MS m / z: 358 [M+H] + , 1 H NMR (600 MHz, CDCl3) δ 6.65 (s, 1H), 4.36 (s, 1H), 3.71 (s, 3H), 3.70 (s, 3H), 3.19 (hept, J = 6.6 Hz, 1H), 2.77 (d, J = 12.6 Hz, 1H), 2.29 (td, J = 13.8, 4.2 Hz, 1H), 1.94 (dd, J = 13.2, 3.6 Hz, 2H), 1.78–1.70 (m, 1H), 1.62–1.54 (m, 2H), 1.44 (d, J = 13.2 Hz, 1H), 1.21–1.15 (m, 4H), 1.11 (d, J = 7.2 Hz, 3H), 1.07 (d, J = 7.2 Hz, 3H), 0.82 (s, 3H), 0.76 (s, 3H). 13 C NMR (150 MHz, CDCl3) δ 177.39, 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.

[0283] Synthesis of Compound II-7-2: Under nitrogen protection, add Compound II-6-1 (550 mg, 1.67 mmol) and 1,4-dioxane (10 mL) to a 25 mL single-necked flask, stir to dissolve, then slowly add DDQ (400 mg, 1.76 mmol), stir at room temperature for 30 min, and monitor the reaction completion by TLC. Filter the reaction solution under reduced pressure, wash the filter cake with dichloromethane (10 mL), evaporate the filtrate to dryness under reduced pressure, and purify the residue by silica gel column chromatography (dichloromethane - methanol, volume ratio 100:1) to obtain 515 mg of dark green solid (II-7-2), with a yield of 93.6%. ESI-MS m / z: 329 [M+H] + , 1 H NMR (600 MHz, CDCl3) δ 8.19 (s, 1H), 6.59 (s, 1H), 4.23 (s, 1H), 2.84 (hept, J = 6.6 Hz, 1H), 2.60 (d, J = 14.4 Hz, 1H), 2.18 (td, J = 13.8, 4.2 Hz, 1H), 1.96–1.90 (m, 1H), 1.90–1.77 (m, 2H), 1.60–1.50 (m, 2H), 1.42 (d, J = 13.2 Hz, 1H), 1.14 (td, J = 13.8, 3.6 Hz, 1H), 1.01 (dd, J = 10.8, 7.2 Hz, 6H), 0.78 (d, J = 12.6 Hz, 6H). 1313C NMR (150 MHz, CDCl3) δ 179.94, 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. Synthesis of Compounds II-8-1 to II-8-4: Under nitrogen protection, add 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) to a 25 mL single-necked flask, stir to dissolve, heat to 110 °C, reflux for 10 h, and monitor the reaction by TLC until completion. Cool to room temperature, evaporate acetic acid under reduced pressure, add saturated sodium bicarbonate solution (20 mL) to the residue, extract with ethyl acetate (20 mL × 3), add anhydrous sodium sulfate to the extract for drying, filter, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (dichloromethane - methanol, volume ratio 150:1 to 80:1) to obtain 40 mg of mixture a and 76 mg of mixture 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. 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 1H NMR (600 MHz, CDCl3) δ 8.02 (s, 1H), 7.34 (d, J = 4.8 Hz, 1H), 7.03 (s, 1H), 4.68 (ddd, J = 5.4, 3.6, 2.4 Hz, 1H), 3.57 (hept, J = 6.6 Hz, 1H), 3.01 (d, J = 13.8 Hz, 1H), 2.37 (td, J = 13.2, 4.2 Hz, 1H), 2.18–2.06 (m, 2H), 1.92–1.85 (m, 1H), 1.75 (dt, J = 13.8, 3.6 Hz, 1H), 1.65 (dd, J = 10.2, 5.4 Hz, 1H), 1.58 (d, J = 13.2 Hz, 1H), 1.33 (dd, J = 13.2, 6.6 Hz, 6H), 1.28 (dd, J = 13.8, 3.6 Hz, 1H), 0.96 (s, 3H), 0.85 (s, 3H). 1313C NMR (150 MHz, CDCl3) δ 177.65, 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. Compound II-8-2: White solid, ESI-MS m / z: 361 [M+Na] + , 1 1H NMR (600 MHz, CDCl3) δ 8.07 (s, 1H), 7.03 (s, 1H), 6.65 (d, J = 4.8 Hz, 1H), 4.65 (ddd, J = 5.4, 3.6, 2.4 Hz, 1H), 3.34 (hept, J = 6.6 Hz, 1H), 3.10 (d, J = 13.8 Hz, 1H), 2.69 (td, J = 13.8, 4.2 Hz, 1H), 2.16–2.07 (m, 2H), 1.87 (t, J = 11.4 Hz, 1H), 1.80 (dp, J = 13.8, 3.6 Hz, 1H), 1.66 (dd, J = 10.2, 5.4 Hz, 1H), 1.56 (d, J = 13.2 Hz, 1H), 1.34 (dd, J = 6.6, 6.0 Hz, 6H), 1.31–1.24 (m, 1H), 0.96 (s, 3H), 0.85 (s, 3H). 13 13C NMR (150 MHz, CDCl3) δ 177.70, 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.

[0285] Compound II-8-3: White solid, ESI-MS m / z: 432 [M+Na] + , 11H NMR (600 MHz, CDCl3) δ 8.02 (s, 1H), 7.09 (s, 1H), 6.88 (q, J = 7.8, 6.6 Hz, 1H), 5.08 (dd, J = 3.6, 2.4 Hz, 1H), 4.72 (qd, J = 13.8, 6.6 Hz, 2H), 3.57 (hept, J = 6.6 Hz, 1H), 2.96 (d, J = 13.8 Hz, 1H), 2.37 (td, J = 13.2, 4.2 Hz, 1H), 2.15–2.02 (m, 2H), 1.89 (s, 3H), 1.84 (ddd, J = 12.6, 10.4, 2.4 Hz, 1H), 1.72 (dt, J = 13.8, 3.6 Hz, 1H), 1.62 (dd, J = 10.2, 5.4 Hz, 1H), 1.55 (d, J = 13.2 Hz, 1H), 1.35 (t, J = 7.2 Hz, 6H), 0.83 (s, 6H). 13 13C NMR (150 MHz, CDCl3) δ 175.52, 171.34, 151.34, 145.12, 140.37, 139.59, 138.15, 123.07, 115.21, 59.50, 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.

[0286] Compound II-8-4: White solid, ESI-MS m / z: 432 [M+Na] + , 1 1H NMR (600 MHz, CDCl3) δ 8.07 (s, 1H), 7.08 (s, 1H), 6.66 (s, 1H), 5.04 (dd, J = 3.6, 2.4 Hz, 1H), 4.75 (qd, J = 13.8, 6.6 Hz, 2H), 3.34 (hept, J = 6.6 Hz, 1H), 3.05 (d, J = 15.0 Hz, 1H), 2.70 (td, J = 13.8, 4.2 Hz, 1H), 2.14–2.00 (m, 2H), 1.88 (s, 3H), 1.85–1.76 (m, 3H), 1.61 (dd, J = 10.2, 5.4 Hz, 1H), 1.54 (d, J = 13.2 Hz, 1H), 1.34 (dd, J = 7.2, 3.6 Hz, 6H), 0.83 (d, J = 7.8 Hz, 6H). 1313C NMR (150 MHz, CDCl3) δ 175.91, 171.22, 151.91, 147.87, 139.18, 135.95, 131.23, 130.06, 116.54, 59.38, 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.

[0287] Example 5: Synthesis of Salvianol 6-position Modified Derivatives

[0288]

[0289] Synthesis of Compound III-1: Add Salvianol I-2 (5.0 g, 15.10 mmol) into a 150 mL round-bottom flask. Under nitrogen protection, add 50 mL of acetone and stir to dissolve. Then slowly add potassium carbonate (6.3 g, 45.65 mmol), and dropwise add methyl iodide (12.9 g, 90.90 mmol). After dropping, heat to 65 °C and reflux for 48 h. After TLC detection shows that the reaction is complete, filter to remove the solid. Wash the filter cake with 20 mL of dichloromethane, evaporate the solvent, add 50 mL of ethyl acetate and 50 mL of saturated ammonium chloride solution to the residue, shake and separate the layers. Extract the aqueous phase with 50 mL of ethyl acetate twice. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether:ethyl acetate = 80:1) to obtain a colorless oil III-1 (5.08 g, yield 90.40%). 1 1H NMR (600 MHz, Chloroform-d) δ 6.67 (s, 1H), 6.36 (dd, J = 9.6, 3.0 Hz, 1H), 6.03 (dd, J = 9.6, 3.0 Hz, 1H), 3.78 (s, 3H), 3.75 (s, 3H), 3.71 (d, J = 13.2 Hz, 1H), 3.53 (s, 3H), 3.24 (p, J = 7.2 Hz, 1H), 2.55 (t, J = 3.0 Hz, 1H), 1.81 (qt, J = 13.8, 3.6 Hz, 1H), 1.64 (dp, J = 14.4, 3.7 Hz, 1H), 1.55 (td, J = 13.2, 3.6 Hz, 1H), 1.47 (d, J = 13.2 Hz, 1H), 1.29 (td, J = 13.2, 4.2 Hz, 1H), 1.21 (d, J = 7.2 Hz, 3H), 1.18 (d, J = 7.2 Hz, 3H), 1.01 (s, 3H), 0.88 (s, 3H). 1313C NMR (150 MHz, Chloroform-d) δ 174.27, 152.86, 150.56, 141.71, 131.91, 130.22, 127.90, 127.08, 120.49, 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.

[0290] 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 protection, 30 mL of acetonitrile was added and stirred until dissolved, then 30 mL of water was added. The temperature was lowered to 0 °C, and after stirring for 10 min, m-chloroperoxybenzoic acid (2.1 g, 12.10 mmol) was added, and stirring was continued at 0 °C for 2 h. After the reaction was completed as detected by TLC, the acetonitrile was evaporated under reduced pressure. 30 mL of saturated sodium bicarbonate solution was added, and the aqueous phase was extracted 3 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 1H NMR (600 MHz, Chloroform-d) δ 7.18 (s, 1H), 4.50 (dd, J = 11.4, 8.4 Hz, 1H), 4.44 (d, J = 8.4 Hz, 1H), 3.74 (s, 3H), 3.66 (d, J = 5.4 Hz, 6H), 3.36 (dt, J = 14.4, 7.8 Hz, 1H), 3.25 (p, J = 7.2 Hz, 1H), 1.90–1.82 (m, 1H), 1.78 (d, J = 11.4 Hz, 2H), 1.57–1.50 (m, 1H), 1.36–1.31 (m, 1H), 1.25 (s, 3H), 1.22 (dd, J = 11.8, 7.2 Hz, 6H), 0.97 (s, 3H). 13 13C NMR (150 MHz, Chloroform-d) δ 176.84, 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.

[0291] 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 protection, 30 mL of methanol was added and stirred at room temperature until dissolved. Sodium hydroxide (600 mg, 15 mmol) was added, and stirring was continued at room temperature for 1 h. After TLC detection showed that the compound reaction was complete, 1 mL of acetic acid was added to neutralize 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, liquid separation was carried out. 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%. 1 H 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=13.8,4.2Hz,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.

[0292]

[0293] Synthesis of Compound Ⅲ-4-1: Add Compound Ⅲ-3 (200 mg, 0.53 mmol) to a 25 mL round-bottom flask. After displacing the air with nitrogen, add 10 mL of dry dichloromethane and pyridine (125 mg, 1.60 mmol). Lower the temperature of the system to 0 °C. After stirring for 10 min, slowly add benzoyl chloride (150 mg, 1.06 mmol). After the addition is complete, slowly raise the temperature of the system to room temperature and stir at room temperature for 24 h. After detecting that the reaction is complete by TLC, add 10 mL of dilute hydrochloric acid (1 mol / L) and stir for 30 min to quench the reaction. Add 20 mL of dichloromethane for dilution and 20 mL of dilute hydrochloric acid (1 mol / L). Separate the organic phase by liquid separation. Then wash the organic phase with 30 mL of saturated sodium bicarbonate. After liquid separation, add anhydrous sodium sulfate for drying. After filtration, evaporate the solvent under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to obtain 150 mg of a white solid with a yield of 58.60%. 1 H 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=12.0Hz,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.

[0294]

[0295] Synthesis of Compound Ⅲ-4-2: Add nicotinic acid (72 mg, 0.58 mmol), N,N-diisopropylethylamine (165 mg, 1.32 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (302.30 mg, 0.79 mmol) into a 25 mL reaction flask. Under nitrogen protection, add 10 mL of dry DMF. After stirring at room temperature for 0.5 h, add Compound Ⅲ-3 (200 mg, 0.53 mmol) and continue stirring for 8 h. After detecting the disappearance of Compound 43 by TLC, add 50 mL of water and 20 mL of saturated brine, cool to 0 °C, and a yellow solid precipitates. Filter to collect the solid. Add 30 mL of dichloromethane to the solid residue to dissolve it, add anhydrous sodium sulfate for drying, filter, evaporate the solvent under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain 94 mg of a white solid with a yield of 36.72%. 1 H NMR (600 MHz, Chloroform-d) δ 8.85 (d, J = 2.4 Hz, 1H), 8.74 (dd, J = 4.8, 1.7 Hz, 1H), 8.05 (dt, J = 7.8, 1.8 Hz, 1H), 7.33 (dd, J = 8.4, 5.4 Hz, 1H), 6.81 (s, 1H), 5.80–5.68 (m, 1H), 5.52 (d, J = 4.8 Hz, 1H), 3.86 (s, 3H), 3.84 (s, 3H), 3.29 (p, J = 7.2 Hz, 1H), 2.86 (d, J = 15.6 Hz, 1H), 2.50 (td, J = 13.8, 4.2 Hz, 1H), 1.97 (qt, J = 13.8, 3.0 Hz, 1H), 1.71 (dd, J = 12.0, 3.6 Hz, 2H), 1.34 (td, J = 13.8, 3.6 Hz, 1H), 1.13 (d, J = 6.6 Hz, 3H), 1.05–1.00 (m, 6H), 0.95 (s, 3H).

[0296] Example 6: Experiment on the alleviation of atrophy of mouse myoblasts (C2C12) induced by the supernatant of mouse colon cancer cell (C26) culture solution by salvigenol derivatives

[0297] The diameter measurement method was used to measure the diameter of myotubes after the differentiation of mouse myoblasts (C2C12). The staining method used in the experiment was hematoxylin-eosin staining, abbreviated as HE staining. The hematoxylin staining solution is alkaline and positively charged, and it can easily bind to the negatively charged and acidic deoxyribonucleic acid (DNA) in the cell nucleus by ionic bonds and stain blue; eosin is an acidic dye, which dissociates into negatively charged anions in water and can easily bind to the positive charges of the amino groups in the proteins in the cytoplasm and stain red. The stained cells were sampled under a high-power microscope, and the diameter of myotube cells was statistically analyzed using Digimizer software.

[0298] The specific method is as follows: Inoculate C2C12 cells at 30,000 / mL in a 24-well plate and differentiate them into mature myotube cells using high-glucose DMEM culture medium containing 2% HS + 1% PS. In addition, inoculate C26 cells and 3T3-L1 adipocytes in T75 flasks respectively, and the culture system is high-glucose DMEM culture medium containing 10% FBS + 1% PS, and place them in a 5% CO2, 37 °C cell culture incubator. When subculturing, subculture at 6 million cells / flask. After culturing in 20 mL of culture medium for 48 h, take the supernatant, centrifuge at 1000 rpm for 3 min, take the supernatant, centrifuge at 4000 rpm for 10 min, and take the supernatant. Mix the 3T3-L1 supernatant and 2% HS differentiation solution at a ratio of 1:1 as the healthy control group. Mix the C26 supernatant and 2% HS differentiation solution at a ratio of 1:1 as the muscle atrophy induction solution. Add an equal amount of muscle atrophy induction solution to both the model group and the drug administration group. At the same time, add different stock solutions of carnosol and carnosol derivatives at the following concentrations of 12.5 μM or 25 μM to the cells, as shown in Table 1.

[0299] Table 1 is the drug administration plan for the experiment of reversing muscle atrophy of carnosol and carnosol derivatives on C2C12 muscle cells in Example 6

[0300] Table 1. Drug administration samples for the atrophy experiment of myoblasts (C2C12) in Example 6

[0301]

[0302]

[0303] Among them, μM refers to μMol / L.

[0304] The method for measuring the diameter of myotubes is as follows:

[0305] After the drug acts for 48 h, fix it with a fixing solution (absolute ethanol: formaldehyde: glacial acetic acid = 20:2:1) for more than 1 h, stain it using hematoxylin-eosin staining method, and collect images under a high-power microscope. Use Digimizer to statistically analyze the diameter of myotubes.

[0306] Calculate the muscle atrophy reversal rate according to the following formula:

[0307] Muscle atrophy reversal rate = (average myotube value of the drug administration group - average myotube value of the model group) / (average myotube value of the control group - average myotube value of the model group) × 100%

[0308] Results and conclusions: Please refer to the appendix Figures 1 - 54 Appendix Figures 1 - 54 is a representative HE staining picture of salvigenin and salvigenin derivatives relieving C26 cell culture solution-induced atrophy of C2C12 mature myotubes. Table 2 in the appendix is the myotube statistical result table. As shown in the appendix Figures 1 - 54 and Table 2, salvigenin derivatives have an obvious reversing effect on muscle cell atrophy.

[0309] Table 2 is the statistical result of salvigenin and salvigenin derivatives relieving atrophy of C2C12 muscle cells, corresponding to the appendix Figures 1 - 54 .

[0310] Table 2. Muscle atrophy reversal rate of C2C12 myotubes under the action of salvigenin and salvigenin derivatives

[0311]

[0312]

[0313] Example 7: Experiment on the effect of I-4-6, II-6-2, II-8-1 and II-8-2 on the viability of mouse myoblasts (C2C12)

[0314] The MTT method was used to detect the survival rate of C2C12 cells. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline formazan and deposit it in the cells, while dead cells do not have this function. The triple solution (10% SDS + 5% isobutanol + 0.01% concentrated hydrochloric acid, aqueous solution) can dissolve the formazan in the cells. The optical absorption value is measured at a wavelength of 570 nm with an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystal formation is proportional to the number of cells.

[0315] The effect of the drug on the viability of mouse myoblasts (C2C12) can be evaluated by the above method. The specific method is as follows:

[0316] C2C12 cells were seeded in a 24-well plate at a density of 30,000 cells / mL. The culture medium was high-glucose DMEM supplemented with 10% FBS and 1% PS, and the cells were incubated in a cell culture incubator with 5% CO2 at 37°C. When the cell confluence reached 70% - 80%, the culture medium was replaced with a differentiation medium containing 2% HS and 1% PS in high-glucose DMEM. The differentiation medium was changed every 48 h, and myotube differentiation was matured on the 5th or 6th day. Compounds Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 or Ⅱ-8-2 were dissolved in the differentiation medium and added at the following concentrations: 1.0625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, and 100 μM, and the mature C2C12 myotubes were treated for 48 h. The culture medium in the plate was discarded, 200 μL of fresh DMEM medium without FBS was added to each well, and 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 triple solution was added to each well and incubated overnight, and then the absorbance was measured at a wavelength of 570 nm using a microplate reader.

[0317] Cell viability (%) = (OD drug - OD blank ) / (OD control - OD blank ) × 100%

[0318] Results and conclusions: Please refer to Appendix Figures 55 - 58 , Appendix Figures 55 - 58The survival rates of C2C12 myotubes after 48 hours of treatment with different concentrations of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 are as follows. As shown in Table 3, when the concentration of Ⅰ-4-6 is 1.0625 μM, the survival rate is 105.959%; when the concentration is 3.125 μM, the survival rate is 102.148%; when the concentration is 6.25 μM, the survival rate is 93.721%; when the concentration is 12.5 μM, the survival rate is 107.047%; when the concentration is 25 μM, the survival rate is 103.457%; when the concentration is 50 μM, the survival rate is 102.811%; when the concentration is 100 μM, the survival rate is 98.752%. When the concentration of Ⅱ-6-2 is 1.0625 μM, the survival rate is 101.057%; when the concentration is 3.125 μM, the survival rate is 98.091%; when the concentration is 6.25 μM, the survival rate is 102.223%; when the concentration is 12.5 μM, the survival rate is 103.822%; when the concentration is 25 μM, the survival rate is 101.669%; when the concentration is 50 μM, the survival rate is 69.800%; when the concentration is 100 μM, the survival rate is 11.228%. When the concentration of Ⅱ-8-1 is 1.0625 μM, the survival rate is 101.778%; when the concentration is 3.125 μM, the survival rate is 108.838%; when the concentration is 6.25 μM, the survival rate is 109.828%; when the concentration is 12.5 μM, the survival rate is 102.572%; when the concentration is 25 μM, the survival rate is 104.249%; when the concentration is 50 μM, the survival rate is 98.067%; when the concentration is 100 μM, the survival rate is 7.763%. When the concentration of Ⅱ-8-2 is 1.0625 μM, the survival rate is 105.227%; when the concentration is 3.125 μM, the survival rate is 109.616%; when the concentration is 6.25 μM, the survival rate is 110.236%; when the concentration is 12.5 μM, the survival rate is 101.260%; when the concentration is 25 μM, the survival rate is 106.090%; when the concentration is 50 μM, the survival rate is 49.165%; when the concentration is 100 μM, the survival rate is 8.176%.

[0319] The above results indicate that the maximum safe concentration of Ⅰ-4-6 for C2C12 muscle cells is 100 μM or higher; the maximum safe concentration of Ⅱ-6-2 for C2C12 muscle cells does not exceed 25 μM; the maximum safe concentration of Ⅱ-8-1 for C2C12 muscle cells does not exceed 50 μM; the maximum safe concentration of Ⅱ-8-2 for C2C12 muscle cells does not exceed 25 μM.

[0320] Table 3 shows 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 Attachment Figures 55 - 58 .

[0321] Table 3. Survival rate of C2C12 cells under the action of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 at different concentrations

[0322]

[0323]

[0324] Example 8 Experiment on the alleviation of atrophy of mouse myoblasts (C2C12) induced by the supernatant of mouse colon cancer cells C26 by Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0325] The myotube diameter after the differentiation of mouse myoblasts (C2C12) was measured by the diameter measurement method. The staining method used in the experiment was hematoxylin-eosin staining method, abbreviated as HE staining method. Hematoxylin staining solution is alkaline and positively charged, and it can easily bind to the negatively charged and acidic deoxyribonucleic acid (DNA) in the cell nucleus by ionic bonds and stain blue; eosin is an acidic dye, which dissociates into negatively charged anions in water and can easily bind to the amino positive charges of proteins in the cytoplasm and stain red. The stained cells were sampled under a high-power microscope, and the Digimizer software was used to statistically analyze the myotube cell diameter.

[0326] The protein immunoblotting experiment was used to evaluate the changes in the levels of related proteins in mouse myoblasts (C2C12). The protein samples separated by PAGE (polyacrylamide gel electrophoresis) were transferred to a solid-phase carrier (such as nitrocellulose membrane). The solid-phase carrier adsorbed the protein by non-covalent bonds and could maintain the polypeptide types separated by electrophoresis and their biological activities unchanged. The protein or polypeptide on the solid-phase carrier was used as an antigen to react with the corresponding antibody, and then react with the enzyme or isotope-labeled secondary antibody. After substrate color development or autoradiography, the protein components expressed by the specific target gene separated by electrophoresis were detected. Through the above method, the effect of salvigenin analogs on the protein level of the C2C12 cell myotrophy model can be evaluated.

[0327] The specific method is as follows: Inoculate C2C12 cells at a density of 30,000 cells / mL in a 24-well plate and differentiate them into mature myotubes using high-glucose DMEM medium containing 2% HS + 1% PS. Additionally, inoculate C26 cells and 3T3-L1 adipocytes in T75 flasks respectively, with the culture system being high-glucose DMEM medium containing 10% FBS + 1% PS, and place them in a 5% CO2, 37 °C cell culture incubator. When passaging, passage at 6 million cells per flask. After culturing with 20 mL of the culture medium for 48 h, take the supernatant, centrifuge at 1000 rpm for 3 min, take the supernatant, centrifuge at 4000 rpm for 10 min, and take the supernatant. Mix the 3T3-L1 supernatant and the 2% HS differentiation medium at a ratio of 1:1 as the healthy control group. Mix the C26 supernatant and the 2% HS differentiation medium at a ratio of 1:1 as the muscle atrophy induction medium. Add an equal amount of the muscle atrophy induction medium to both the model group and the administration group. At the same time, add the stock solution of I-4-6 to the cells at the following concentrations: 6.25 μM, 12.5 μM, 25 μM, as shown in Table 4; add the stock solution of II-6-2 at 6.25 μM, 12.5 μM, 25 μM, as shown in Table 4; add the stock solution of II-8-1 at 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, as shown in Table 4. Add the stock solution of II-8-2 at 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, as shown in Table 4.

[0328] Table 4 shows the dosing regimens 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.

[0329] Table 4. Drug samples for the atrophy experiment of myoblasts (C2C12) in Example 8

[0330] Serial number Number Experimental sample Component 1 CT Control group 1 2% HS differentiation solution 2 C26 medium Model group 1 2% HS differentiation solution + C26 supernatant 3 C26 + Ⅰ - 4 - 6 (6.25 μM) Experimental group 1 2% HS differentiation solution + C26 supernatant + Ⅰ - 4 - 6 (6.25 μM) 4 C26 + Ⅰ - 4 - 6 (12.5 μM) Experimental group 2 2% HS differentiation solution + C26 supernatant + Ⅰ - 4 - 6 (12.5 μM) 5 C26 + Ⅰ - 4 - 6 (25 μM) Experimental group 3 2% HS differentiation solution + C26 supernatant + Ⅰ - 4 - 6 (25 μM) 6 C26 + Ⅱ - 6 - 2 (6.25 μM) Experimental group 4 2% HS differentiation solution + C26 supernatant + Ⅱ - 6 - 2 (6.25 μM) 7 C26 + Ⅱ - 6 - 2 (12.5 μM) Experimental group 5 2% HS differentiation solution + C26 supernatant + Ⅱ - 6 - 2 (12.5 μM) 8 C26 + Ⅱ - 6 - 2 (25 μM) Experimental group 6 2% HS differentiation solution + C26 supernatant + Ⅱ - 6 - 2 (25 μM) 9 CT Control group 2 2% HS differentiation solution 10 C26 medium Model group 2 2% HS differentiation solution + C26 supernatant 11 C26 + Ⅱ - 8 - 1 (3.125 μM) Experimental group 7 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 1 (3.125 μM) 12 C26 + Ⅱ - 8 - 1 (6.25 μM) Experimental group 8 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 1 (6.25 μM) 13 C26 + Ⅱ - 8 - 1 (12.5 μM) Experimental group 9 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 1 (12.5 μM) 14 C26 + Ⅱ - 8 - 1 (25 μM) Experimental group 10 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 1 (25 μM) 15 C26 + Ⅱ - 8 - 2 (3.125 μM) Experimental group 11 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 2 (3.125 μM) 16 C26 + Ⅱ - 8 - 2 (6.25 μM) Experimental group 12 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 2 (6.25 μM) 17 C26 + Ⅱ - 8 - 2 (12.5 μM) Experimental group 13 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 2 (12.5 μM) 18 C26 + Ⅱ - 8 - 2 (25 μM) Experimental group 14 2% HS differentiation solution + C26 supernatant + Ⅱ - 8 - 2 (25 μM)

[0331] Among them, μM refers to μmol / L.

[0332] The method for measuring myotube diameter is as follows:

[0333] After the drug acts for 48 h, fix with a fixative (absolute ethanol: formaldehyde: glacial acetic acid = 20:2:1) for more than 1 h, stain using hematoxylin-eosin staining method, and collect images under a high-power microscope. Use Digimizer to statistically analyze the myotube diameter.

[0334] Calculate the muscle atrophy reversal rate according to the following formula:

[0335] Muscle atrophy reversal rate = (average myotube value of the administration group - average myotube value of the model group) / (average myotube value of the control group - average myotube value of the model group) × 100%

[0336] The method for Western blotting is as follows:

[0337] Protein sample preparation: After the cells were treated with the drug for 48 h, the cell culture medium in the 6-well plate was removed, and the cells were washed 3 times with pre-warmed PBS at 37 °C. After complete digestion with trypsin, the digestion was terminated with a culture medium containing 10% FBS + 1% PS. The cell suspension was centrifuged at 4 °C and 1000 rpm for 5 min, washed once with ice-cold PBS, the supernatant was discarded, and then centrifuged again for 3 min to discard the supernatant. RIPA containing 1% phosphatase inhibitor was added in proportion, and vortexed every 10 min for a total of 30 min. The whole lysis process was carried out on wet ice. After lysis, the lysate was centrifuged at 4 °C and 12,000 rpm for 10 min, and the supernatant was taken. The protein supernatant was diluted 5-fold (i.e., 5 μL of protein supernatant was added to 20 μL of ultrapure water). In addition, protein standards with 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 with ultrapure water, and an ultrapure water control was set as the zero-adjustment well, with a total volume of 25 μL. The BCA working solution was prepared at a ratio of A solution: B solution = 50:1. 200 μL of the BCA working solution and 25 μL of the protein dilution were added to each group, and reacted in the dark at 37 °C for 30 min. Then, the OD value was measured at 562 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and the protein concentration of the sample was calculated using the standard curve. The loading amount was set at 20 μg, and the loading system was 20 μL. The protein was aliquoted according to the protein concentration and dilution factor, heated and denatured in a metal bath at 100 °C for 10 min, cooled on ice, and stored at -80 °C after centrifugation or directly loaded onto the gel.

[0338] Protein electrophoresis: Prepare an appropriate gel concentration and Marker according to the target molecular weight. After the gel solidifies, place it in an electrophoresis tank containing 1x electrophoresis buffer. Pull out the comb and remove the bubbles, then load the samples in order. The samples need to be vortexed evenly before loading. First, run the gel at a constant voltage of 70 V until the samples are compressed into a line and the Marker has started to separate, and then change the voltage to 110 V until the Marker with a molecular weight of 25 KD runs out of the bottom edge of the gel.

[0339] Transfer membrane: Prepare an appropriate amount of transfer buffer. Activate a PVDF membrane with a suitable size in anhydrous methanol for 30 s, and then place the membrane in the prepared transfer buffer. Gently pry open the two glass plates in the middle, cut off the useless parts, and carefully remove the gel. According to the rule of black gel and white membrane and the sandwich method, clamp them in the order of black clamping plate - sponge - three layers of filter paper - gel - PVDF membrane - three layers of filter paper - sponge - white clamping plate in turn, and remove the bubbles in each step. Place it in the electrophoresis tank, connect the power supply, and transfer the membrane at a constant current of 250 mA on ice for 1 h.

[0340] Immune response: After the transfer was completed, the PVDF membrane was cut according to the position of the target band and placed in 5% milk prepared with TPBS. It was blocked at room temperature on a side-shaking bed for more than 1 h. The blocking solution was removed, and the bands were washed three times with TPBS for 10 min each time. 5% milk prepared with TPBS was used as the primary antibody dilution solution, and the primary antibody was diluted at a ratio of 1:1000 and incubated overnight at 4°C with 1 mL. The primary antibody was removed, and the bands were washed three times with TPBS for 10 min each time. 5% milk prepared with TPBS was used as the secondary antibody dilution solution, and the corresponding secondary antibody was diluted at a ratio of 1:5000. The bands were incubated on a side-shaking bed at room temperature for 1 - 2 h, and the bands were washed three times with TPBS for 10 min each time. Solution A and Solution B in the ECL kit were mixed at a ratio of 1:1, evenly applied to the target protein, and after appropriate reaction, the membrane was placed in the Amersham Imager600 machine to detect chemiluminescence and take pictures.

[0341] Results and conclusions: Please refer to the appendix Figures 59 - 78 . Appendix Figures 59 - 76 shows the representative HE staining pictures of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 alleviating C26 cell culture medium-induced atrophy of C2C12 mature myotubes, appendix Figure 77 , Figure 78 is the statistical result graph of myotubes, and Table 6 in the appendix is the statistical result table of myotubes. Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 have an obvious reversing effect on myocyte atrophy, and show a concentration-dependent relationship. When the concentration of Ⅰ-4-6 is 6.25 μM, the reversal rate is 21.53%; when the concentration is 12.5 μM, the reversal rate is 61.39%; when the concentration is 25 μM, the reversal rate is 67.08%. When the concentration of Ⅱ-6-2 is 6.25 μM, the reversal rate is 28.40%; when the concentration is 12.5 μM, the reversal rate is 43.02%; when the concentration is 25 μM, the reversal rate is 66.55%. When the concentration of Ⅱ-8-1 is 3.125 μM, the reversal rate is 42.17%; when the concentration is 6.25 μM, the reversal rate is 35.55%; when the concentration is 12.5 μM, the reversal rate is 42.63%; when the concentration is 25 μM, the reversal rate is 73.21%. When the concentration of Ⅱ-8-2 is 3.125 μM, the reversal rate is 44.05%; when the concentration is 6.25 μM, the reversal rate is 58.19%; when the concentration is 12.5 μM, the reversal rate is 63.15%; when the concentration is 25 μM, the reversal rate is 88.52%.

[0342] Table 5 shows the statistical results of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 alleviating atrophy of C2C12 muscle cells, corresponding to appendix Figures 59 - 78 .

[0343] Table 5. Reversal rate of myotube atrophy of C2C12 under the action of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2

[0344]

[0345]

[0346] Appendix Figures 79 - 82 It is the result of the immunoblot experiment on the protein level effects of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 on the C2C12 cell muscle atrophy model. According to the results, it is found that the C26 supernatant not only activates p-p65 in C2C12 myotubes, leading to overexpression of the E3 ubiquitin ligase Atrogin-1 and reduced expression levels of MHC and MyoD, but also inhibits the phosphorylation of AKT. On the one hand, Ⅰ-4-6, Ⅱ-8-1 or Ⅱ-8-2 can not only inhibit the expression of p-p65 in a concentration-gradient manner, thereby inhibiting the overexpression of Atrogin-1 and promoting the expression of MHC and MyoD; at the same time, Ⅰ-4-6, Ⅱ-8-1 or Ⅱ-8-2 inhibit the reduction of AKT phosphorylation level caused by the 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 upregulates the reduction of AKT phosphorylation level caused by the C26 supernatant in a concentration-dependent manner.

[0347] The above results indicate that Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 can relieve protein degradation in muscle cells, promote myocyte differentiation and growth, and have an obvious alleviating effect on muscle cell atrophy through a new molecular mechanism of action, and show a concentration-dependent relationship.

[0348] Example 9 Experiment on the effects of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 on the viability of mouse preadipocytes (3T3-L1)

[0349] The MTT method was used to detect the survival rate of 3T3-L1 cells. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline formazan and deposit it in the cells, while dead cells do not have this function. The triple solution (10% SDS + 5% isobutanol + 0.01% concentrated hydrochloric acid, aqueous solution) can dissolve the formazan in the cells, and the optical absorbance value is measured at a wavelength of 570 nm with an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystal formation is proportional to the number of cells.

[0350] The effects of drugs on the viability of mouse preadipocytes (3T3-L1) can be evaluated by the above method. The specific method is as follows:

[0351] The 3T3-L1 cells were seeded in a 24-well plate at a density of 30,000 cells / mL. The culture medium was high-glucose DMEM containing 10% FBS + 1% PS, and the cells were incubated in a cell culture incubator with 5% CO2 at 37°C. After the cells reached confluence and continued to fuse for 6 days, they were differentiated into mature adipocytes using high-glucose DMEM medium containing 0.5 mM IBMX, 5 mg / mL insulin, 1 μM dexamethasone, and 10% FBS. After successful differentiation, a large number of lipid droplets were clearly visible in the cells. Compounds I-4-6, II-6-2, II-8-1, or II-8-2 were added at the following concentrations: 1.0625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, and 100 μM, and were formulated in high-glucose DMEM medium containing 10% FBS and then applied to the differentiated and mature 3T3-L1 adipocytes for 48 h. The culture medium was replaced with a solution of MTT (5 mg / mL): culture medium = 1:10, 200 μL / well. After 4 h, 50 μL of the triple solution was added to each well and incubated overnight. After overnight incubation, the absorbance was measured at a wavelength of 570 nm using a microplate reader.

[0352] Cell viability (%) = (OD drug - OD blank ) / (OD control - OD blank ) × 100%

[0353] Results and conclusions: Please refer to Appendix Figures 83 - 86 . Appendix Figures 83 - 86are the survival rates of 3T3-L1 cells under the action of different concentrations of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1, and Ⅱ-8-2 for 48 h. As shown in Table 7, when the concentration of Ⅰ-4-6 is 1.0625 μM, the survival rate is 95.898%; when it is 3.125 μM, the survival rate is 98.395%; when the concentration is 6.25 μM, the survival rate is 101.607%; when the concentration is 12.5 μM, the survival rate is 104.963%; when the concentration is 25 μM, the survival rate is 94.231%; when the concentration is 50 μM, the survival rate is 91.439%; when the concentration is 100 μM, the survival rate is 99.025%. When the concentration of Ⅱ-6-2 is 1.0625 μM, the survival rate is 107.97%; when the concentration is 3.125 μM, the survival rate is 113.401%; when the concentration is 6.25 μM, the survival rate is 115.701%; when the concentration is 12.5 μM, the survival rate is 95.76%; when the concentration is 25 μM, the survival rate is 100.736%; when the concentration is 50 μM, the survival rate is 108.566%; when the concentration is 100 μM, the survival rate is 99.726%. When the concentration of Ⅱ-8-1 is 3.125 μM, the survival rate is 104.226%; when the concentration is 6.25 μM, the survival rate is 94.599%; when the concentration is 12.5 μM, the survival rate is 95.514%; when the concentration is 25 μM, the survival rate is 92.752%; when the concentration is 50 μM, the survival rate is 92.876%; when the concentration is 100 μM, the survival rate is 71.851%. When the concentration of Ⅱ-8-2 is 3.125 μM, the survival rate is 100.707%; when the concentration is 6.25 μM, the survival rate is 103.165%; when the concentration is 12.5 μM, the survival rate is 96.889%; when the concentration is 25 μM, the survival rate is 97.478%; when the concentration is 50 μM, the survival rate is 92.876%; when the concentration is 100 μM, the survival rate is 80.929%.

[0354] The above results indicate that the maximum safe concentration of Ⅰ-4-6 for 3T3-L1 adipocytes is 100 μM or above; the maximum safe concentration of Ⅱ-6-2 for 3T3-L1 adipocytes is 100 μM or above; the maximum safe concentration of Ⅱ-8-1 for 3T3-L1 adipocytes does not exceed 50 μM; the maximum safe concentration of Ⅱ-8-2 for 3T3-L1 adipocytes does not exceed 50 μM.

[0355] Table 6 shows 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 attached Figures 83 - 86 .

[0356] Table 6. Survival rates of 3T3-L1 cells under the action of different concentrations of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0357]

[0358]

[0359] Experimental results of the alleviation of lipolysis in 3T3-L1 adipocytes by Ⅰ-4-6 and Ⅱ-6-2 in Example 10

[0360] The intracellular fat content was evaluated by Oil Red O fat staining method. Oil Red O is a fat-soluble dye that can be highly dissolved in fat and can specifically stain neutral fats such as triglycerides in tissues.

[0361] The changes in the levels of related proteins in mouse adipocytes (3T3-L1) were evaluated by Western blot assay.

[0362] The effect of the drug on the cell lipolysis model can be evaluated by the above methods. The specific methods are as follows: 3T3-L1 cells were seeded in 24-well plates at a density of 30,000 cells / mL, and the culture system was high-glucose DMEM medium containing 10% FBS + 1% PS, and placed in a 5% CO2, 37 °C cell culture incubator. After the cells grew to confluence and continued to fuse for 6 days, they were differentiated into mature adipocytes with high-glucose DMEM medium containing 0.5 mM IBMX, 5 mg / mL insulin, 1 μM dexamethasone and 10% FBS. After successful differentiation, a large number of oil droplets were clearly visible in the cells. In addition, C26 cells and C2C12 myotubes were seeded in T75 flasks respectively. After growing to confluence, the medium was replaced with fresh phenol red-free high-glucose DMEM medium. After 48 h, the phenol red-free high-glucose DMEM supernatant of C26 cells was collected, centrifuged at 1000 rpm for 3 min, and the supernatant was taken. Then it was centrifuged at 4000 rpm for 10 min, and the supernatant was taken. The supernatant of C2C12 cells and phenol red-free high-glucose DMEM medium were mixed at a ratio of 1:1 as the healthy control group. The supernatant of C26 and phenol red-free high-glucose DMEM medium were mixed at a ratio of 1:1 as the lipolysis inducer. The model group and the drug administration group were both added with an equal amount of lipolysis inducer. At the same time, the Ⅰ-4-6 stock solution was added to the cells at the following concentrations: 6.25 μM, 12.5 μM, 25 μM, as shown in Table 7. The Ⅱ-6-2 stock solution was added to the cells at the following concentrations: 6.25 μM, 12.5 μM, 25 μM, as shown in Table 7.

[0363] Table 7 shows the drug administration plan for the lipolysis reversal experiment of Ⅰ-4-6 on 3T3-L1 adipocytes in Example 10.

[0364] Table 7. Drug administration samples for the lipolysis experiment of mouse preadipocytes (3T3-L1) in Example 10

[0365]

[0366] The Oil Red O staining and semi - quantitative method are as follows: After the cells are treated with the drug for 48 h, dilute 0.5% Oil Red O dye (prepared with isopropanol) with distilled water at a ratio of 3:2, mix well, and filter with a 0.42 - μm filter head to obtain the staining solution. Discard the cell culture medium, fix the cells with 4% neutral formaldehyde for more than 1 h, suck out the fixing solution with a vacuum pump, wash the cells 3 times with PBS, place them in a well - ventilated area at room temperature to dry for 20 min, add the Oil Red O staining solution to stain for 30 min and then remove it, wash the cells 2 times with PBS and then wash away the floating color with 60% isopropanol, wash 2 times with distilled water, and then observe and take pictures under an inverted high - power microscope. Then dissolve the Oil Red O bound in the cells with isopropanol, and perform parallel operations with the undifferentiated adipocyte staining group as the blank zero - adjustment group. The staining group performs parallel operations as the blank zero - adjustment group, and measure the OD value at 510 nm with an enzyme - linked immunosorbent assay (ELISA) reader. According to the OD value results, perform Oil Red O semi - quantification according to the following formula:

[0367] Fat content (%)=(OD drug - OD blank ) / (OD control - OD blank )×100%

[0368] The Western blotting method is the same as that described in Example 8.

[0369] Results and conclusions: Please refer to Appendix Figures 87 - 98 . Appendix Figures 87 - 96 is a representative picture of Oil Red O staining for Ⅰ - 4 - 6 and Ⅱ - 6 - 2 to relieve the lipolysis of 3T3 - L1 mature adipocytes induced by C26 supernatant. In the normal control group, a large number of lipid droplets accumulate in the cells, the lipid droplets are larger, and the coloring is deep and bright; after the C26 supernatant acts alone, the lipid droplets in the cells become significantly smaller and the content decreases sharply; when the C26 supernatant is incubated with Ⅰ - 4 - 6 or Ⅱ - 6 - 2 simultaneously, as the concentration of Ⅰ - 4 - 6 or Ⅱ - 6 - 2 increases, the content of lipid droplets stained with Oil Red O in the cells gradually increases and the lipid droplets become larger.

[0370] Appendix Figure 97 , 98 is the semi - quantitative result of Oil Red O staining. As shown in Figure 97 , 98, Ⅰ - 4 - 6 and Ⅱ - 6 - 2 increase the intracellular Oil Red O content in a concentration - dependent manner, indicating an increase in fat content.

[0371] Appendix Figure 99It is the result of the Western blot experiment of protein in the lipolysis model of 3T3-L1 cells by Ⅰ-4-6. The results show that under the stimulation of C26 supernatant, p-p65 is activated in 3T3-L1 adipocytes, and its expression level is up-regulated. Ⅰ-4-6 decreases the expression of p-p65 in a concentration-dependent manner; C26 cell culture medium activates the phosphorylation of HSL (hormone-sensitive lipase), and the expression level of p-HSL is up-regulated, participating in the lipolysis process. As the concentration of Ⅰ-4-6 increases, the expression of p-HSL decreases; C26 cell culture medium activates the phosphorylation of AMPKα (AMP-dependent protein kinase α subtype), and the expression level of p-AMPKα is up-regulated, and energy metabolism is intensified. As the concentration of Ⅰ-4-6 increases, the expression of p-AMPKα decreases.

[0372] The above results indicate that Ⅰ-4-6 has an obvious alleviating effect on adipocyte lipolysis by a new molecular mechanism, reducing fat degradation and alleviating excessive energy consumption, and shows a concentration-dependent relationship.

[0373] PK experiments of Ⅰ-5-3, Ⅰ-4-22, Ⅰ-4-6, Ⅱ-6-2 and Ⅱ-8-1 in Example 11

[0374] BALB / c mice were injected with Ⅰ-5-3, Ⅰ-4-22, Ⅰ-4-6, Ⅱ-6-2 or Ⅱ-8-1 via the tail vein, and the solvent was 5% DMSO + 1% HS-15 + 94% 0.9% Saline, and the dose was 25 mg / kg for all.

[0375] Results and conclusions: Please refer to Appendix Figure 100 and Table 8. The AUC 0-t (ng·hr / mL), T 1 / 2 (hr) of Ⅰ-5-3, Ⅰ-4-22, Ⅰ-4-6, Ⅱ-6-2 and Ⅱ-8-1 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, 273 ng·hr / mL and 1.03 h respectively. The area under the blood drug concentration curve (AUC 0-t ), drug half-life (T 1 / 2 ) and drug residence time (MRT) of Ⅰ-4-6 and Ⅱ-6-2 were relatively ideal.

[0376] Table 8 shows the PK experiment results of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2 in Example 11, corresponding to Appendix Figure 100 Table 8. Parameter data of PK experiments of Ⅰ-4-6, Ⅱ-6-2, Ⅱ-8-1 and Ⅱ-8-2

[0377]

[0378] Experimental Results of Treating Tumor Cachexia Animal Model with Example 12 Ⅰ-4-6 and Ⅱ-6-2

[0379] Pre-inoculate the C26 cell suspension with 1.5 million cells in each axilla of BALB / c mice for preservation. When the tumor volume increases to 1000 cm 3 , take out the tumor, homogenize it with 0.5 mL PBS / g to obtain a tumor tissue suspension, and count and dilute it to 1 million / 100 μL. The mice to be inoculated are grouped by body weight, and the cell suspension is inoculated into the left axilla of BALB / c mice at a dose of 100 μL / mouse. Ⅰ-4-6 and Ⅱ-6-2 are dissolved into a uniform and stable solution with 3% DMSO, preheated 2% Ethanol, 1% HS-15, and 94% 0.9% Saline solution, with a final concentration of 4 mg / mL, a dosage of 40 mg / kg, and an administration route of intraperitoneal injection, and administered on the 3rd day after inoculation. Monitor the body weight, body temperature, tumor size, and food intake of the mice every day. When the body weight of the mice in the model group decreases by about 15% or the tumor volume reaches about 2000 mm 3 at about the 16th day, it is considered to enter the advanced stage of cachexia. After decapitating and sacrificing the mice, biochemical samples such as gastrocnemius muscle, epididymal fat, tumor, and serum are obtained.

[0380] Appendix Figures 101 - 105 shows the tumor-bearing body weight, tumor-free body weight, tumor volume, tumor weight, and tumor anatomical diagram during the survival period of the mice. As shown in the figure, the body weight of the healthy group of mice continued to increase; the tumor-bearing body weights of the mice in the Ⅰ-4-6, Ⅱ-6-2 groups, and the C26 tumor model group continued to rise from the start of the experiment to the 11th day, and began to decline sharply after the 11th day until the end of the experiment, and the same was true for the tumor-free body weight; while the reduction of the tumor-bearing body weight and tumor-free body weight in the Ⅰ-4-6 group was relatively gentle, and the tumor-bearing body weight and tumor-free body weight were higher than those in the C26 Model group from the 14th to the 16th day, and the differences were statistically significant (p < 0.5).

[0381] Appendix Figures 106 - 107 shows the gastrocnemius muscle mass and the actual picture results of the gastrocnemius muscle. There was no difference in the gastrocnemius muscle mass between the Ⅰ-4-6 group and the C26 tumor model group, and there was no statistical significance in terms of mass.

[0382] Appendix Figures 108 - 109 shows the epididymal fat mass and the actual picture results of the epididymal fat. The epididymal fat mass in the Ⅰ-4-6 group of epididymal fat was significantly greater than that in the C26 tumor model group, and the difference was statistically significant (p < 0.5).

[0383] The above results show that Ⅱ-6-2 cannot relieve the weight loss, muscle atrophy, and fat degradation caused by tumor cachexia without affecting the tumor size. Ⅰ-4-6 can relieve the weight loss caused by tumor cachexia by inhibiting fat degradation without affecting the tumor size.

[0384] The above is the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the technical field covered by the present invention, without departing from the method of the present invention, several supplements and improvements can still be made, but these supplements and improvements should also be regarded as the protection scope of the present invention.

Claims

1. Salvianolic acid compounds, characterized in that, Comprising a compound represented by formula (X) or a pharmaceutically acceptable salt of the compound represented by formula (X), Formula (X); In the said formula (X), A is O; B is O; C is N; When A is O, R1 is selected from H, a substituted or unsubstituted 5-7-membered aromatic heterocyclic group, phenyl, a substituted or unsubstituted C1-C 12 alkyl group; the substituents are selected from one or more of halogen, C1-C6 alkyl, cyano, trifluoromethyl, nitro, carboxyl, hydroxyl, hydroxymethyl, methoxy, amino, acetylamino, methanesulfonyl, methoxyamide group; the aromatic heterocyclic group contains one or more heteroatoms selected from N, O, S.

2. Salvianolic acid compounds, characterized in that, The structural formula of the compound is or 。 3. Use of the carnosol compound according to claim 1 or 2 in the preparation of a medicament for treating cachexia diseases.

4. A pharmaceutical composition, characterized in that, The said composition contains the carnosol compound according to claim 1 or 2, and / or any one or more pharmaceutically acceptable excipients.

5. Use of the pharmaceutical composition according to claim 4 in the preparation of a medicament for treating cancer cachexia diseases.

6. The application according to claim 5, wherein, The product dosage form of the said pharmaceutical composition is a capsule, tablet, oral preparation, microcapsule preparation, injection, ointment, spray or suppository; the product administration method of the said pharmaceutical composition is injection, oral administration, parenteral, inhalation spray or transdermal administration.

7. The application according to claim 5 or 6, characterized in that, The said cancer cachexia is muscle atrophy caused by tumor tissue; and / or, the said cancer cachexia is fat reduction caused by tumor tissue; and / or, the said cancer cachexia is reduced appetite caused by tumor tissue.

8. The application according to claim 7, characterized in that, The said tumor is a solid tumor.

9. The application according to claim 7, characterized in that The said cancer cachexia is cancer cachexia caused by digestive tract-related cancers, liver cancer and lung cancer.

10. The application according to any one of claims 5, 6, 8 or 9, characterized in that The carnosol compound represented by formula (X) accounts for 0.001-100 wt.% of the total dry weight of the composition.

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